Modern

BIOTECHNOLOGY
Modern
BIOTECHNOLOGY
Panacea or new Pandora’s box?

Johannes Tramper and Yang Zhu

Wageningen Academic
P u b l i s h e r s
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 CONTENTS

PREFACE 13

PART ONE: INTRODUCTION -- MODERN BIOTECHNOLOGY: PANACEA OR NEW PANDORA’S BOX? 17

1. MODERN BIOTECHNOLOGY: A BLESSING OR A CURSE? 19
1.1. What is (modern) biotechnology? 20
1.2. Bioethics 20
Textbox 1.1. Structure and function of genetic material. 21
1.3. Bioterrorism 25
1.4. Recombinant DNA technology 25
Textbox 1.2. A “triple lock” on the door! 26
1.5. Biotechnology debate 27
1.6. Sources 28
2. MODERN BIOTECHNOLOGY: FOOD FOR DISCUSSION! 29
2.1. The history in a nutshell 30
Textbox 2.1. The recombinant DNA technology. 31
Textbox 2.2. Herman, the transgenic bull. 32
2.2. Supporters and opponents 34
Textbox 2.3. The transgenic tomato “Flavr Savr”. 34
2.3. Why transgenic plants? 37
2.4. Why transgenic animals? 40
Textbox 2.4. Dolly the clone. 41
2.5. Gene and stem cell therapy 43
2.6. EU Legislation 45
2.7. Conclusion 47
2.8. Sources 48
3. GENETICALLY MODIFIED CROPS AND THE EUROPEAN UNION 49
3.1. Introduction 50
Textbox 3.1. Vatican: GM not against God’s will. 51
3.2. Seven focal points to accept GM crops within the EU 52
Textbox 3.2. The 20-year environmental safety record of GM trees. 52
Textbox 3.3. Modern biotechnology: scientific victim? 53
Textbox 3.4. The precautionary principle. 56
Textbox 3.5. Statistics as a framework for decision making. 58
3.3. Conclusions 63
3.4. Sources 64

7
 PART TWO: OUR DAILY FOOD AND DRINK 69
4. CHEESE: BIOTECHNOLOGY THROUGH THE AGES 71
4.1. Old cheese 72
4.2. Traditional curdling 72
Textbox 4.1. Cheese alliance. 73
4.3. Modern curdling 73
Textbox 4.2. The PluGbug technology. 75
4.4. Cheese ripening: now and in the future 76
Textbox 4.3. Ripening agents. 77
Textbox 4.4. The NICE system. 78
4.5. The final question 78
Textbox 4.5. Acceptability of genetically modified cheese. 79
4.6. Sources 80
5. BIOTECHNOLOGY IN THE BAKERY: ON THE RISE! 81
5.1. Our daily bread 82
Textbox 5.1. Sourdough. 82
5.2. Baker’s yeast 83
5.3. Dough 83
Textbox 5.2. Baker’s yeast. 83
5.4. Bread improvers 84
5.5. Enzymes 84
Textbox 5.3. Acrylamide reduction. 85
5.6. Recombinant enzymes 86
Textbox 5.4. Bread enzymes are also good for the environment. 87
5.7. Transgenic crops 88
Textbox 5.5. AMFEP’s policy declaration on modern biotechnology. 89
5.8. Legislation 91
5.9. In conclusion 93
5.10. Sources 94
6. WINE: ONE OF THE OLDEST BIOTECHNOLOGICAL PRODUCTS 95
6.1. What is wine? 96
6.2. The first wine 96
Textbox 6.1. The amphora and traditional Greek wine: retsina. 97
6.3. Alcohol as a stimulant 98
6.4. The scientific discoverer: Louis Pasteur 98
6.5. How is wine made? 99
6.6. Enzymes are the solution! 100
6.7. Champagne with a flick of the wrist 101
6.8. Manipulation of wine yeast 102

8
 Textbox 6.2. GM yeasts: the next battleground? 104
6.9. Manipulation of the grapes 106
Textbox 6.3. GM Grapes Raise Hopes for Midwest Wine Industry. 107
6.10. Winemakers raise their glasses tobiotechnology 108
6.11. In conclusion 110
6.12. Sources 111
7. MEAT FROM THE BIOTECH VAT 113
7.1. Scope 114
7.2. Animal feed 114
7.3. Growth hormones 118
Textbox 7.1. Recombinant gelatin. 120
7.4. Meat processing 121
7.5. Cloned meat 123
7.6. New developments 125
7.7. Biotechnological meat substitutes 127
Textbox 7.2. ‘Happy Birthday’. 127
7.8. In conclusion: Happy Meat! 128
7.9. Sources 130
8. “FRANKENFOOD” 131
8.1. Food and genes 132
8.2. Justified fears? 133
8.3. Are GM foods harmful to health? 134
Textbox 8.1. Risk assessment of GMOs. 135
Textbox 8.2. Genetically modified rice fights allergies. 137
8.4. More anxiety! 139
8.5. Who is telling the whole truth? 140
Textbox 8.3. Golden Rice. 141
8.6. Is there a future for transgenic crops? 144
Textbox 8.4. Chopping onions without tears. 145
8.7. Conclusions 146
8.8. Sources 148

PART THREE: HEALTH HAS LIMITS 151

9. ANTIBIOTICS 153
9.1. Antibiotics: life-saving biotechnology 154
9.2. The bacteria fight back 156
Textbox 9.1. Phases in drug development. 157
9.3. The prospects 159
Textbox 9.2. Vicissitudes of a typical anti-infectives biotech company. 162

9
 9.4. ‘Green’ production 164
9.5. A never-ending story 165
9.6. Taking another look at phages 166
9.7. Conclusions 167
9.8. Sources 169
10. HORMONES: NATURAL REGULATORS 171
10.1. What are hormones? 172
10.2. Human growth hormone (HGH) 173
10.3. Erythropoietin (EPO) 178
Textbox 10.1. The French EPO test. 180
Textbox 10.2. Operación Puerto file still open. 182
10.4. Puregon™: follicle-stimulatinghormone (FSH) 184
10.5. In conclusion 186
10.6. Sources 187
11. GENE THERAPY: A PANACEA FOR GENETIC ABNORMALITIES? 189
11.1. What is gene therapy? 190
11.2. A short history of gene therapy 192
Textbox 11.1. The Gelsinger case. 194
Textbox 11.2. Glybera: gene therapy for lipoprotein-lipase deficient patients. 196
11.3. SCID children 197
11.4. Gene therapy in the uterus 199
11.5. Not everything can be treated (yet) 200
Textbox 11.3. Genetics in a nutshell. 201
11.6. Gene doping 202
Textbox 11.4. The German muscleman. 202
Textbox 11.5. Hormone mafia becomes gene mafia. 204
11.7. Gene therapy: not yet a panacea or a revolution 205
11.8. Sources 206
12. XENOTRANSPLANTATION 207
12.1. The history of xenotransplantation: a shocking past 208
Textbox 12.1. The first xenotransplantation. 208
Textbox 12.2. Human rejuvenation transplants. 210
12.2. The transgenic ‘spare-part pig’ 212
Textbox 12.3. The immune system – some basic facts. 214
Textbox 12.4. Dysregulated coagulation in pig-to-primate xenotransplantation. 216
12.3. Pandemic risks 218
12.4. Social and ethical aspects 219
12.5. In conclusion 223
Textbox 12.5. Willem Kolff. 224

10
 Textbox 12.6. The “perfect match”! 225
12.6. Sources 226
13. THE HUMAN GENOME PROJECT 227
13.1. The human genome 228
12.2 ‘The book of life’ 228
13.3. Human genome sequencing 231
13.4. A new paradigm in health care 231
Textbox 13.1. Diary of Karen Rich; 1 April 2059. 233
13.5. Will the Netherlands climb on the bandwagon? 234
13.6. The surprises of the genome 235
13.7. Where are we now? 236
Textbox 13.2. Paradigm shift: one gene = one protein → one gene = several proteins. 237
Textbox 13.3. The Personal Genome Project. 240
13.8. In conclusion 242
Textbox 13.4. ‘Reading genes’. 242
13.9. Sources 245
14. STEM CELL THERAPY: PROMISING AND CONTROVERSIAL! 247
14.1. Human embryonic stem cells are ‘hot’ 248
14.2. From Bush to Obama 249
14.3. The controversies 251
14.4. What is a stem cell (therapy)? 254
14.5. Types of stem cells 255
14.6. The making of human (embryonic) stem cell lines 257
Textbox 14.1. Pre-implantation genetic diagnostics. 259
Textbox 14.2. Human embryos cloned. 261
14.7. Formation of induced human embryonic pluripotent stem cells by dedifferentiation 263
14.8. In conclusion 265
Textbox 14.3. Summary of key ISSCR guidelines for the translation of stem cell research into the clinic. 266
14.9. Sources 267

PART FOUR: EPILOGUE: CASSANDRA 269

15. MODERN BIOTECHNOLOGY: FOR BETTER OR FOR WORSE? 273

INDEX 277

11
 PREFACE

There’s a long history behind the writing of this book. At its McElroy in a really pleasant collaboration with us. We then
roots are many lectures on modern biotechnology given produced an updated and international version. Where
by one of us (JT) over the last two decades, and still being desirable and appropriate, we replaced typical Dutch cases
delivered at universities and schools, in libraries, for service by international ones and in doing so removed most of
clubs, etc. Many times the question “Why don’t you put it the references to Dutch journals, daily newspapers, etc.
all in a book?” was raised. This suggestion solidified in the However, the English text still benefits greatly from pieces
first half of 2001 when JT took a sabbatical leave at EPFL in of text from Dutch (popular) science writers. We refer to our
Lausanne (Switzerland). A rudimentary draft of the preceding Dutch book for their credits.
Dutch version of this book was written at that time. In the present book the number of (complex) links to
For various reasons this first draft lay pretty well untouched websites is further increased. To facilitate visiting them they
until 2007. In that year the contents of our jobs changed, are numbered and the direct links can easily be found on the
which allowed us to work structurally on an update and website of the publisher: www.wageningenacademic.com/
in November 2009 the Dutch version was published. modernbiotech.
Somebody who greatly facilitated the completion of the book We gratefully acknowledge the grant from the Netherlands
is Tim Jacobs, a young creative graphical designer, who Biotechnology Foundation and the large order by the
prepared all the figures, cartoons, strips, etc. The regular Netherlands Genomics Initiative; it allowed us to print the
meetings with him forced us to stay on track and were in fact book in full colour. We would also like to acknowledge the
a great joy. This continued to be the case when he prepared pleasant and professional collaboration with Wageningen
new graphics for this book. Academic Publishers; having the publisher next door is very
Thanks to grants from Wageningen University and the handy.
Wageningen University Fund we were able to widely Finally, we should finish with a long list of names of those
distribute the Dutch book in the Netherlands. Soon people who, over the years, have contributed in one way or another
started to ask: Why didn’t you write it in English? Our answer to the eventual publication of this book. However, the risk
was this: It is born out of the Dutch situation, but we are now of forgetting somebody is so great that we decided simply
working on an international version written in English. to issue the following statement: our sincerest thanks to
With another grant from Wageningen University, a everyone!
professional translation was first carried out by Sandra The authors

13
part one
Introduction
 MODERN BIOTECHNOLOGY:
PANACEA OR NEW PANDORA’S BOX?

In Greek mythology Pandora is the ‘giver of all’ or the ‘all endowed’ and the first mortal woman to be sent to earth
upon the orders of Zeus. She was given a mysterious box, which she was forbidden to open. Pandora, however, not
only possessed the charm and beauty of a goddess, a gift from Aphrodite, she was also very curious, a characteristic
given her by the god Hermes. Once on earth, therefore, she was unable to resist taking a look inside the box. It was
filled with gifts and calamities, all of which, to her dismay, escaped and spread throughout humanity, with all the
disastrous consequences thereof. Only the spirit of hope was left at the bottom. Figuratively speaking, Pandora’s
box is a source of much suffering. Is modern biotechnology a Pandora’s box, as anti-biotechnology movements
would have us believe or is it a panacea to cure many of the world’s ills? Therein lies the pivotal question in this
book. Our final conclusion is that biotechnology can be the source of much good if it is handled wisely; in other
words, we should lift the lid of this new Pandora’s box carefully and with discretion.

PANDORA’S BOX...

IR
SPA
DE
E
DISEAS

A STER
DI S
HOPE

DORA
PAN

17
1 MODERN BIOTECHNOLOGY:
A BLESSING OR A CURSE?

Developments in the area of modern biotechnology can no longer be stopped. Take, for example, the amazing
pace at which our knowledge and understanding of the genetic material of humans (Textbox 1.1) is moving and the
possibilities that this opens up for health care and forensic science. It’s vital to put this into practice in a sensible
and controlled manner. Winning the trust of the public must be the first step. But reliable information and continuous
communication are crucial if that is to happen. In this book we aim to go some way towards achieving this. The
main focus is on the more controversial topics, such as gene therapy versus gene doping, or therapeutic versus
reproductive cloning. The most famous example of cloning is Dolly the sheep, born in 1996 and the first cloned
mammal. In this chapter we aim to make just a passing acquaintance with modern biotechnology for those who
are unfamiliar with this fast-evolving area of expertise. We have tried to write the various chapters so that they can
stand alone and be read separately. The textboxes contain more detailed information, basic knowledge, or typical
examples, but are not needed for understanding the main body of the text.

THE BIRTH OF DOLLY WAS NOTHING SHORT OF A MIRACLE...

BUT VERY SHORT ON ROMANCE!
BLAM
M
OO
B
KA

J. Tramper, Y. Zhu, Modern Biotechnology – Panacea or new Pandora’s box? 19

DOI 10.3920/978-90-8686-725-7_1, © Wageningen Academic Publishers 2011
 1.1. WHAT IS (MODERN) BIOTECHNOLOGY? opponent while Jimmy Carter, ex-president of the
US, and winner of the 2002 Nobel Peace Prize as
There are many definitions of biotechnology. Basically well as founder of the non-profit Carter Center1, is a
they all come down to the same thing: biotechnology major supporter. The Vatican gives a cautious nod
is the integration of life sciences with engineering. to biotechnology, on the understanding that there
The production of semi-synthetic antibiotics like should continue to be a ban on cloning humans and
amoxicillin by using moulds and enzymes is an ‘tinkering’ with human DNA. It is also clear that even
excellent example of this. People talk about modern renowned scientists cannot seem to agree with each
biotechnology when recombinant DNA technology other and nobody can guarantee absolute safety.
is involved, also called gene technology or genetic The burning question is whether biotechnology
modification (see below and also Textbox 2.1 in the and its products are more dangerous than more
next chapter). Opponents of modern biotechnology conventional equivalents and whether they fit into
consistently use the term genetic manipulation the picture we as a society have of the future. For
because it provokes a negative association. And yet us to be able to establish this, we must keep up
gene technology doesn’t attract universal criticism. the societal debate and continue to research and
Quite the opposite. There are many examples of develop modern biotechnology.
great new products of modern biotechnology and
there are many more in the pipeline, especially in 1.2. BIOETHICS
the medical domain. The development of genetically
modified moulds has, for instance, made the At the beginning of 2002, Francis Fukuyama’s book Our
production of antibiotics far more efficient in the Posthuman Future: Consequences of the Biotechnology
last few decades. Since mid-1990, however, there Revolution (Fukuyama, 2002) was discussed in many
have been heated discussions among supporters book review sections of newspapers, magazines and
and opponents of modern biotechnology, especially journals, e.g. Abrams (2003) and Spier (2002). Francis
in Europe. The thorny issues in these debates have Fukuyama is certainly not a run-of-the-mill thinker. He is
been estimating the risks in terms of health and a renowned political philosopher, and at the beginning of
the environment. It seems that, for the time being this century – as a member of the presidential advisory
at least, it won’t be easy to close the gap that has council on bioethics – he directly advised George Bush
opened up over the years between the various points on matters such as cloning, the use of embryonic stem
of view. The European debate is a good reflection of cells and genetic selection and modification. His opinions
what is happening around the world. For example, therefore partly determined how the Bush government
Prince Charles of Great Britain is an out-and-out
1
www.cartercenter.org

20 Part 1: Introduction
TEXTBOX 1.1.
Structure and function of genetic material.

Genetic information is stored in DNA molecules

(deoxyribonucleic acid). A DNA molecule is a long
strand of nucleotides which are linked to each other
by phosphate groups (the black balls in Figure 1.1).
A nucleotide consists of a deoxyribose molecule (the
sugar ribose in which an –OH is replaced by –H) to
which a nitrogen base is attached. DNA contains four
different nitrogen bases: adenine (A), cytosine (C),
guanine (G) and thymine (T). Genetic characteristics
are established as genes in the DNA molecules. A
gene is a piece of a DNA molecule that codes for a
specific protein. In other words, if a cell contains DNA
with a specific gene, this cell can theoretically make
(express) the protein encoded by this gene. Proteins
regulate all processes in the living cell and as such are Figure 1.1. The two-dimensional structure of DNA.
the building blocks of life. They themselves are made
up of 20 smaller units, called amino acids. and these function like dimmer switches, regulating
The sequence of the nucleotides is the code used the action of genes or groups of genes. In addition,
to lay down genetic information. This code is always between the genes there are much bigger pieces of
laid down in sets of three building blocks, the so- DNA whose function we do not yet know, and which
called triplet code (Figure 1.2). One triplet is called have long been thought to have no function at all.
a codon and represents one amino acid in the amino This view, however, is increasingly being challenged.
acid chain from which proteins are then produced. These pieces are still frequently, and unjustly, called
Most amino acids have several codons and there are junk DNA.
also stop codons. This doesn’t mean that the whole The protein synthesis is carried out by ribonucleic
DNA molecule is used from start to finish to code acid (RNA). This consists of a single stranded chain,
genetic information. On the contrary, these codes are similar in structure to that of DNA, the difference
distributed in small packages on the DNA, the genes. being that deoxyribose and thymine are replaced by
Other pieces of DNA are located between the genes, ribose and uracil (U), respectively. RNA is made in the

Chapter 1: Modern biotechnology: a blessing or a curse? 21

 cell nucleus on the DNA (transcription) and occurs molecule with a specific triplet of unpaired bases, the
in three forms. Messenger-RNA (mRNA) takes the anticodon. This pairs with the corresponding codon
information necessary for the protein synthesis from in the mRNA molecule in the ribosome, resulting in
the DNA in the nucleus to the protein factories of the the coupling of amino acids to each other to form
cell, the so-called ribosomes. Transfer-RNA (tRNA) proteins according to the base sequence in the mRNA
is the form which transports amino acids to the (translation). In most organisms ribosomes roughly
ribosomes and sequences them along the mRNA. consist of equal parts of protein and ribosomal RNA,
For each amino acid there is a separate tRNA the third form of RNA.

Helix construction

Nucleotide A T G C
building blocks Adenine Thymine Guanine Cytosine

Double
DNA helix

Information flow
TCT GTG
ACT TCA
TTA ACT CCG
AGA CAC
TGA AGC AGT
Part of AAT GTA TGA GGC
ATT
gene TCG CAT
TAA

Transcription
UCU GUG
ACU UCA
UUA ACU CCG
Part of AGC GUA
mRNA AUU

Thr
Ser Translation

Part of Val Thr

Val Ser
protein Pro
Leu
eu Ile
Ser

Figure 1.2. The protein synthesis (Thr, Ser, Val, etc. are the separate amino acids in the protein chains).

22 Part 1: Introduction
dealt with modern biotechnology. One example of his should be limited to therapeutic purposes. Jordaan
influence was the ban imposed by the Bush government therefore submits that Fukuyama’s main argument
on using state funding for embryonic stem cell research must be rejected. Subsequently he analyses the
(see Chapter 14). According to Fukuyama, state power supporting arguments in Our Posthuman Future –
should be used to lay down the rules for biotechnology, relating to reproductive freedom and human rights,
and this should not be left to science or business, nor social justice, and psychology – and concludes that
individual freedom of choice. The consequences of an none of these arguments can support Fukuyama’s
unlimited application would, in his view, be too drastic contention that new reproductive technologies
and dangerous. should be limited to therapeutic purposes. Jordaan
A critical analysis of Fukuyama’s bioethics was ends his thirteen-page analysis with the paragraph
published by Jordaan (2009). Jordaan identifies four Antipromethean Heresy:
distinct weaknesses in Fukuyama’s main argument, “Our Posthuman Future is a good dose of feel-
i.e. human nature, which is defined as species-typical good drugs in a philosophical sugar-coating for a
genetic characteristics, is the ultimate basis for human bioconservative audience. To a more open-minded,
values, specifically for our species’ special value – our ethically informed audience, Our Posthuman Future
human dignity. Fukuyama infers from this first premise is a macabre effort to resurrect the discredited
that should any aspect of human nature be changed naturalistic fallacy back into mainstream philosophical
by new reproductive technologies, it would endanger discourse after a well-deserved death more than a
not only human values, but also the very basis of century ago … Our Posthuman Future cannot add
human dignity; therefore justifying the limitation of anything to the global bioethics debate – it can only
such new reproductive technologies to therapeutic pollute these already troubled waters with arguments
uses. The four weaknesses Jordaan identifies are: that have the intellectual accountability of a tabloid
(1) Fukuyama’s definition of human nature is vague feature. Dworkin (2002) describes the ethical
and not based on reality; (2) the relationship he posits dimension of the biotechnology revolution as ‘playing
between human nature on the one hand and values with fire’, but states that playing with fire is ‘what we
on the other is weak, or dependent on other, non- mortals have done since Prometheus’. But then,
related values; (3) even accepting his first premise, it not all philosophers can have promethean courage
does not follow that any change in human nature will to face and explore a radically new value paradigm.
necessarily undermine human dignity; and (4) even Fukuyama clearly prefers humankind to live without
accepting his second premise – any change in human this metaphorical fire. The promethean metaphor
nature will necessarily undermine human dignity – it has been a defining paradigm in classical times, as
still does not follow that new reproductive technologies well as in modernity – it was the cultural catalyst

Chapter 1: Modern biotechnology: a blessing or a curse? 23

 for creating the free and technologically advanced Biotechnology is not harmful
contemporary societies of the West … Beware the day
when we betray our promethean heritage. Beware the The article was written by Cynthia Schneider, who at
antipromethean heresy of Fukuyama.” that time was the US ambassador to the Netherlands
Although we are not professional philosophers, we (she left in mid-2001). Schneider wrote this article
subscribe to the view of Jordaan. We believe that to publicise an international conference she had
only open debates by a well-informed public, hand in organised to take place a few days later in The Hague.
hand with ongoing education, scientific research and A symposium attended by a great many of the big
technology development, can create a viable future for shots in the area of modern biotechnology, among
humankind. them J. Craig Venter, who came to champion her
In a leading Dutch newspaper (NRC), two years before proposition. As a result of a false bomb threat issued by
the publication of Fukuyama’s book, the following an opponent of modern biotechnology, the symposium
headline appeared above an article on the opinion was temporarily suspended. We are not opponents
page: of biotechnology. On the contrary, it has been our

BELIEVE ME, BIOTECHNOLOGY IS NOT DANGEROUS!

TIC
TOC

24 Part 1: Introduction
professional field for decades. But neither are we as convincing response! After the terrible attacks on the
decided as Mrs. Schneider. Using the same words, we Twin Towers in 2001, the fear of biological attacks
would like to convert her statement into a question: in particular has continued to increase and many
American citizens have even gone so far as to buy
Isn’t biotechnology harmful? gas masks. More and more countries are therefore
trying to prepare for an attack with biological weapons,
And that is exactly what we will be discussing in this whereby ironically biotechnology itself will probably
book so as to be able to arrive at the final conclusion: also be used to ensure defence. Practically speaking,
Biotechnology doesn’t have to be harmful! If used the research in question often amounts to the same
sensibly, it can be a blessing rather than a curse for thing as developing these weapons. In mid-2007 five
mankind. Or, in the words of Richard Preston, author American laboratories conducting this kind of research
of the bestseller, The Cobra Event (1997): were closed down, because staff there were infected
with offensive pathogens. There is also a suspicion
I don‘t want ‘The Cobra Event’ to be seen as anti- that a lot of defensive work against bio-weapons is
biotechnology or anti-science, since it isn’t. In offensive in nature. For more information on this topic,
the introduction I compare genetic engineering to go to the website set up by the Sunshine Project, an
metallurgy – it can be used to make plowshares or international non-profit organisation that works to bring
swords. The difference is human intent. to light facts about biological weapons2.

1.4. RECOMBINANT DNA TECHNOLOGY

1.3. BIOTERRORISM
The year 1973 was a special year. It was the year of
Preston’s book, sadly enough, is about a terrorist the energy crisis resulting in car-free Sundays. And
attack on New York City. Not with aeroplanes, but with of course, it was also the year of the Watergate affair
a genetically modified virus. The plot rings so true, which led to the resignation of the American president
that former president Bill Clinton asked FBI experts Richard Nixon. But this year is also regarded as the birth
to look into how realistic such an attack was. Opinion of modern biotechnology. It was in 1973 that Stanley
polls in the US, carried out online in 2000, revealed Cohen and Herbert Boyer3, of Stanford University and
that 94% of respondents were worried that their the University of California in Berkeley, carried out the
country was vulnerable to attacks by bioterrorists; 64% first successful recombinant DNA experiments with the
even thought there was a serious risk of the attack
taking place in the first decade of the 21st century. A 2
www.sunshine-project.org
3
web.mit.edu/invent/iow/boyercohen.html

Chapter 1: Modern biotechnology: a blessing or a curse? 25

 TEXTBOX 1.2. Watson, Berg formed a committee to discuss the
A “triple lock” on the door! potential risks of this technology. These discussions
were made public at a conference in Asilomar,
Above it was mentioned that in 1973 Berg, Boyer California, in February 1975, where guidelines for safe
and Cohen were the first to “recombine” an experimentation were laid down. In the first instance,
organism. At the time a discussion these guidelines only concerned microorganisms
was already brewing about the (bacteria, yeasts and moulds). Special instructions for
risks of this new technique. At the plants and animals only came later. The recombinant
request of the National Academy DNA laboratories are literally equipped with locks
of Sciences, Berg sent a letter to and only specially trained researchers are allowed
the journal Science, in which he called to enter. In these laboratories, a vacuum, amongst
for a one-year moratorium on further other things, is supposed to prevent microorganisms
recombinant DNA experiments (Berg from ‘escaping’. ‘Crippled’ microorganisms are used,
et al., 1974) . Together with about 150
4
so that in the event that they do escape, they will not
other scientific experts, including one be able to survive ‘on the outside’. These measures
of the two scientists who discovered the would appear to have been overcautious and have
double helix structure of DNA (Figure 1.2), James since been relaxed.

bacteria Escherichia coli (E. coli). With genetic “copy Since then, the term modern biotechnology has been
and pasting” they made this E. coli, a bacterium that used whenever recombinant DNA technology is applied
lives in our intestines, resistant to two antibiotics, namely (see Textbox 2.1 in Chapter 2 for more information on this
tetracycline and kanamycin. In that same period their technology).
colleague and later Nobel Prize winner Paul Berg (also at The first commercial applications of this technology
Stanford University) modified the genetic material of the followed less than a decade later, in 1982. The Dutch
same microbial strain with a piece of DNA from a cancer- company Intervet was the first on the market with a
inducing virus. These scientists foresaw the enormous vaccine against swine diarrhoea. Hot on their tails was
consequences of this new technology and called for a the American company Eli Lilly with human insulin for
voluntary, temporary moratorium on further research. diabetics, made from genetically modified bacteria (E.
Once guidelines for safe experimentation had been coli). A piece of human DNA - the bit that ensures we
established during the Asilomar conference (Textbox 1.2) can make insulin in our body - was added to the DNA of
in 1975, research in this area took a great leap forward. these bacteria, so that these microorganisms could make
human insulin for us. As a result, unlimited quantities
4
www.pnas.org/cgi/reprint/71/7/2593

26 Part 1: Introduction
of pure human insulin, so to speak, have been made additives, genetically modified plants and animals, and
available at a reasonable price. Furthermore, this insulin cloning. These are discussed in the context of our daily
causes fewer side effects than the old product, i.e. food and drink (Part II) and our health (Part III).
modified swine insulin. There’s no shortage of coverage of gene technology in
the media. While lecturing around and about the country,
1.5. BIOTECHNOLOGY DEBATE however, it has become clear to us that we scientists,
but also those in business, the public sector and the
When recombinant insulin appeared on the market, media, have not yet succeeded in conveying sufficient
there immediately arose a heated debate between knowledge in the area of modern biotechnology to the
supporters and opponents of modern biotechnology. A man in the street. Which is why fear of this technology
German company had developed a similar commercial has sometimes been blown out of all proportion. We are
process at the same time, but only got manufacturing definitely not in favour of indiscriminately implementing
permission from the German government a good ten everything humanly possible with the help of modern
years later due to pressure from the Green Party. The biotechnology. We do, however, believe that wise use
Green Party and other environmental groups forced of gene technology can lead the way in developments
the introduction of very restrictive German legislation that will create new and better products. First and
concerning modern biotechnology, because of the fear foremost it is essential to establish standards and norms
of irreparable damage to the environment and health. for implementing gene technology, such that the man
However, German diabetics protested that they should in the street starts to believe that these developments
also be able to access this new medicine directly. This can proceed safely with no risk to our health and the
led to a hypocritical situation where the product couldn’t environment. With this book we hope to objectively
be manufactured in their own country, while at the same inform the wider public about modern biotechnology in
time it was being imported and sold on the market. The order to reach the point where the discussion can turn
debate about this technology has continued ever since, to the real issues. Not those that are chiefly dictated
and has become even more heated since mid-1990. It by irrational fear and end up in a “yes it is/no it isn’t”
would be no exaggeration to say that we have ended up discussion. But rather, what do we as a society consider
in a situation of trench warfare. In the various chapters to be acceptable risks and which objectives do we
of this book we will be reviewing in particular the more classify as sufficiently important to justify the use of
controversial topics like recombinant products as food modern biotechnology.

Chapter 1: Modern biotechnology: a blessing or a curse? 27

 1.6. SOURCES Fukuyama, F. (2002). Our Posthuman Future: Consequences
of the Biotechnology Revolution. New York, Farrar,
Abrams, F. R. (2003). JAMA, 289(4), 488-490. Straus & Giroux.
Berg, P., Baltimor.D, Boyer, H. W., Cohen, S. N., Davis, R. Jordaan, D. W. (2009). Antipromethean Fallacies: A Critique
W., Hogness, D. S., et al. (1974). Potential biohazards of Fukuyama’s Bioethics. Biotechnology Law Report,
of recombinant DNA-molecules. Science, 185(4148), 28(5), 577-590.
303-303. Preston, R. (1997). The Cobra Event. Toronto, Canada: The
Dworkin, R. (2002). Sovereign virtue: the theory and practice Random House Publishing Group.
of equality (First paperback edition ed.). Cambridge, Spier, R. E. (2002). Toward a New Human Species? Science,
MA, Harvard University Press. 296(5574), 1807-1808.

28 Part 1: Introduction
2 MODERN BIOTECHNOLOGY:
FOOD FOR DISCUSSION!

“It is one thing to have a safe product; it is another to command confidence in the market place”

Stephen Dorrell, former UK Health minister (1995-1997, Conservative party)

Advances in the field of modern biotechnology seem unstoppable now. This view is also expressed in Ernst &
Young’s annual reports on the biotechnology sector. According to their Beyond borders: the global biotechnology
report 20075, biotechnology even experienced a historical leap forward in 2006 with global growth of more than 10
percent. The growth in global areal with transgenic (genetically modified) crops – in the EU the most controversial
issue – has been steady right from the introduction in 1996 (Figure 2.1a) and in 2009 this areal has again grown
by an ample 7%, totalling 134 million hectares (ISAAA6). The growth is especially strong in Brazil, South Africa and
India. Brazil has even overtaken Argentina and is now, after the USA, the largest producer of transgenic crops. This
figure also shows the hesitance with respect to transgenic crops that exists in Europe; the area occupied by these
crops there is marginal (less than 0.1% of the global total). In 2007 this area even halved, which was largely due to
the almost complete disappearance of these crops in Romania (Figure 2.1b). Nevertheless, at the time of writing
this, in the summer of 2010, it seems that even in Europe the tide is also turning in favour of these crops.
130
million ha.

120 US 210
thousand ha.

China
110 Argentina Poland Portugal
Paraguay 180
100 Romania Czech Republic
Brazil Slovakia France
90 South Africa 150 Germany
80 Canada 137.000
70 Uruguay 120
110.000 350 7.146
India
60 Other countries 3.244
50 90 21.147
40 492
60 5.000
30
20 30 53,225 53.667 75.148 79.269 76.057
10
0 0
1996 1997 1998 1999 2000 2001 2002 2003 2004 2005 2006 2007 2008 2009 2005 2006 2007 2008 2009

Figure 2.1. Area occupied by transgenic crop, globally (a) and in Europe (b) . 7

5
www.ey.com/Publication/vwLUAssets/Global_Biotechnology_Report_2007/$FILE/BeyondBorders2007.pdf
6
www.isaaa.org/resources/publications/briefs/41/executivesummary/default.asp
7
www.lisconsult.nl/images/stories/Downloads/arealen%20transgene%20gewassen%201996%20-%202009.pdf

J. Tramper, Y. Zhu, Modern Biotechnology – Panacea or new Pandora’s box? 29

DOI 10.3920/978-90-8686-725-7_2, © Wageningen Academic Publishers 2011
 It is vital that the new advances are put into practice be found in a book by Confucius (500 BC). However,
in a sensible and controlled manner. Gaining the trust what we currently refer to as modern biotechnology
of the wider public, by providing them with objective originated millennia later, i.e. in the second half of the
information, has to be the first step. Knowledge of last century. As mentioned in Chapter 1, scientists
history is useful too. In this chapter we will take a bird’s Paul Berg, Stanley Cohen, Herbert Boyer and their
eye view of the first 35 years of modern biotechnology. co-workers conducted the first successful recombinant
The pertinent legislation is also briefly discussed. DNA experiments in California in 1973 (Textbox 2.1),
thus heralding the advent of modern biotechnology.
2.1. THE HISTORY IN A NUTSHELL In so doing, they ever so slightly lifted the lid of a
new Pandora’s box. In order to ensure that only
Biotechnology is older than documented history. To good things would emerge this time, they continued
give an example, malting and brewing were already with the research only after the guidelines for safe
taking place in Mesopotamia (current-day Iraq) in 4000 experimentation had been established. When this
BC. In China a description of mould growth on grain for happened, two years later, the research progressed in
the production of alcoholic beverages and vinegar can leaps and bounds on a global scale.

STONE AGE BEER DELIVERY

BEER

30 Part 1: Introduction
 TEXTBOX 2.1. what Boyer and Cohen did with their first successful
The recombinant DNA technology. rDNA experiments8. They started from a plasmid
with a gene that caused resistance to the antibiotic
In recombinant DNA (rDNA) tetracycline. Using molecular “cut-and-paste” work they
technology, changes are added a new gene encoding resistance to the antibiotic
The made to the genetic make- kanamycin to this plasmid. They let this recombinant
ACGT up of an organism. Plasmids plasmid be taken up by E. coli. These bacteria later
of play an important role here. A appeared to become resistant to both tetracycline and
life plasmid is a stable, lone, usually kanamycin. Both genes were therefore expressed by
circular piece of DNA, which can the “recombinant” cells, proof that their experiment had
self-replicate in a host cell. It is succeeded. This was done in 1973, the year regarded
therefore not part of the genome of as the dawn of modern biotechnology. Since then, all
a cell. Plasmids are well-known for kinds of techniques have been discovered, for example,
being able to transfer resistance to activating and deactivating (silencing) genes. The end
antibiotics, because they can easily pass is not in sight and the number of areas for application
from one cell to another. And that is exactly seems inexhaustible.

As described in Chapter 1, the first commercial THE SIXTIES AND SEVENTIES WERE
applications of modern biotechnology followed less PRETTY TOUGH FOR PIG BREEDERS
than a decade later in 1982. The Dutch company CRAP, MAN!!!
Intervet was first in line with a recombinant vaccine
against swine diarrhoea. They were closely followed
by the American company Eli Lilly, which manufactured
human insulin by using genetically modified bacteria.
Since then, the genetic modification of microorganisms
(bacteria, yeasts, moulds) has become routine, and
has resulted in a whole range of new spin-off products
on the market, as we shall see later. A further ten years
later, at the end of the ‘80s and beginning of the ‘90s,
genetically modified, or transgenic, animals and plants
came on the scene. The Dutch bull “Herman” and the
American “Flavr Savr” tomato were the trendsetters in
8
web.mit.edu/invent/iow/boyercohen.html

Chapter 2: Modern biotechnology: food for discussion! 31

 TEXTBOX 2.2. immune system, e.g. AIDS patients. The announcement
Herman, the transgenic bull. about it being added also to food for premature babies
generated a lot of fuss (Figure 2.3). Initially, the
The Dutch firm Pharming is the ‘creator’ of the late bull reasoning was that Herman’s female offspring would
Herman, a ‘blaarkop’ (breed of dairy cattle) to whose be better protected against mastitis (udder infection),
genetic material a human gene was added. When and so less antibiotic would be needed to maintain the
Herman was still an embryo, scientists at Pharming health of these transgenic offspring, and there would be
added the human gene that encoded lactoferrin to a resulting reduction in antibiotics in the food chain. After
the DNA in his cells. Human lactoferrin is an infection- many years in the headlines, Herman was put to sleep
inhibiting protein that occurs in substantial quantities on Friday 2 April 2004 by doctors from the Faculty of
in mother’s milk. The immune system of babies is not Animal Health in Utrecht. The board of the Herman Bull
yet sufficiently developed and lactoferrin helps protect Foundation decided on euthanasia because the animal
them against infection. The human lactoferrin gene was was suffering too much from old-age arthritis. The bull,
synthesised by researchers at Pharming and inserted the first genetically modified cattle in the world, was 13
into the cells of the Herman embryo. This was done years old. Herman’s skin is now on display in the Leiden
in the hope that milk from Herman’s female offspring museum, Naturalis. On the Pharming website9 you can
would contain substantial quantities of this substance. read that the company is close to marketing human
The intention was to extract lactoferrin from the milk lactoferrin as an advanced ingredient in nutritional
and market it as a drug for people with a compromised products.

this area (Textbox 2.2 and 2.3). are intended to make the varieties resistant to
Meanwhile, the number of modern biotechnology disease or certain pesticides, or sometimes to allow
applications has multiplied, with a great many still in extra food to be produced, or to enable them to grow
the pipeline. Biotechnology companies and institutes in poor conditions. Golden Rice is an oft-discussed
have introduced new drugs, vaccines, diagnostic example. It is a recombinant variant of rice that
tests, medical treatments, environmentally friendly contains beta carotene, a substance converted to
products and foodstuffs. One of the most spectacular vitamin A in the body. Vitamin A is essential for a
developments has been the cloning of adult mammals; healthy immune system and good eyesight, amongst
“Dolly the sheep” being the now legendary example of other things. This crop is intended to compensate for
this technology (Textbox 2.4). a shortage of vitamin A in the diet of those living in
In addition, genetically modified variants of a great developing countries. This shortage has until now
many crops have been created. These modifications
9
www.pharming.com

32 Part 1: Introduction
 made many people needlessly blind. Meanwhile a
new recombinant variant has been developed that
contains up to 23 times more beta carotene than the
original transgenic variant. More about Golden Rice
in Chapters 3 & 8.

Plant breeding stations have since grown an entire

range of genetically modified plant varieties and
brought them onto the market. In this way corn varieties
(the so-called Bt crops) have been made resistant to
the European corn borer (Ostrinia nubilalis), with the
introduction of a gene that codes for a protein toxin
of the soil-dwelling bacterium Bacillus thuringiensis;
and varieties of corn, cotton, rapeseed and sugar
beet (the H(erbicide) T(olerant) crops) have been
made invulnerable to specific herbicides. As a result
these crops can now be sprayed with those specific
herbicides, for example, Roundup Ready, allowing
for the destruction of competing weeds but not the
plants that are to be harvested. Research carried out
at the University of Illinois shows that growing these
herbicide-resistant crops is better for the environment
than using other techniques for getting rid of weeds
(AgraFood Biotech, 23 July 2007). The US is ahead of
the game in the introduction of these transgenic crops.
As an illustration, more than 90% of the soya bean crop
in that country consists of HT crops. In the same issue
of AgraFood Biotech there is also an article expressing
Figure 2.2. Mother’s milk from cows. Controversial the expectation that, faced with pressure from fast-
poster issued by the Dutch League for Animal Protection rising food prices, acceptance in the European Union
in 1994. We thank this league for providing archived (EU) is also set to increase rapidly. This expectation
material and the approval to use it for publication. was partly based on the increase in the production of

Chapter 2: Modern biotechnology: food for discussion! 33

 transgenic crops in the EU in 2007, mainly in Spain 2.2. SUPPORTERS AND OPPONENTS
and France. However, since then only a decrease has
been witnessed and the areal coverage is extremely At the end of the last century the expectation that
marginal in size compared to countries like the United modern biotechnology would revolutionise agriculture,
States, Argentina, Brazil, India and China. It remains a health care and environmental protection was so great
very difficult issue in the EU. However, as mentioned that, in a cover story in Business Week magazine
above, in mid-2010 it seems that the tide is starting to of 27 July 1998, the 21st century was hailed as the
turn. century of biotechnology10. The introduction of such

TEXTBOX 2.3. Flavr Savr is therefore, as the name would suggest, a

The transgenic tomato “Flavr Savr”. more flavoursome tomato. In 1995 Calgene received
authorisation to sell Flavr Savr in Canada and Mexico.
On 18 May 1994 the FDA declared the transgenic The tomato was sold for a number of years in about
tomato Flavr Savr from the firm Calgene (now 3000 shops under the name MacGregor. It has since
Monsanto) to be as safe as a traditional tomato. This been taken off the market because the yields were
signalled a breakthrough for the first commercial too low. You can read everything there is to know
food product made by using recombinant DNA about Flavr Savr in a book by Belinda Martineau, one
technology. By inserting an extra piece of DNA, Flavr of the researchers at Calgene who worked on the
Savr was genetically modified so that a specific gene development of Flavr Savr. The book is called “First
is not expressed, i.e. the protein encoded by this Fruit. The creation of the Flavr Savr tomato and the
gene is not made by the cell. This process is called birth of biotech food” (Martineau, 2001).
antisense technology. The protein in this case was
polygalacturonase (PG), an enzyme that accelerates
the rotting process. PG breaks down the pectin in the
‘Flavr Savr’
cell walls, so that the tomatoes become soft and rot Tomato
faster. This genetic modification extends the shelf life
of the tomato by at least ten days. It also allows the
tomatoes to ripen fully and turn red on the plant. This
is in contrast to most traditional supermarket tomatoes,
Traditional
which are plucked when green and then treated with Tomato

ethylene gas (especially in the US) to turn them red,

but which have no time to develop taste and aroma.
10
www.businessweek.com/1998/30/b3588002.htm

34 Part 1: Introduction
a spectacular and revolutionary new technology is foodstuffs from the market, and of forcing the FDA
obviously not without its critics. As already mentioned, to test these products more thoroughly and provide
the UK’s Prince Charles is a very public opponent. them with a label before re-releasing them onto the
In the ‘90s he set up a website to stimulate debate market. In October 2000 the judge declared in favour
between supporters and opponents. Prince Charles of the FDA, which was a victory for the American
himself said the following on the subject: “Genetic biotechnology camp and evidence of the stark contrast
engineering takes mankind into realms that belong to in the way we deal with this issue in Europe. The
God and to God alone.” standpoint of the Bush government was, however, far
from unambiguous. On the one hand Bush wanted
to incorporate the development of biotechnology in a
SCIENCE COMPETES WITH RELIGION
Bio-Shield project11 to combat potential bioterror, while
OVER GENETIC ENGINEERING
on the other hand he was attempting to introduce a
total ban on stem cell experiments; his successor
A Barack Obama has a completely different view
G

on the latter (see Chapter 14). The policy of the

T
C Bush government was intended, out of economic
necessity, to vigorously stimulate the export of
genetically modified agricultural products to Europe.
All this makes it clear that, for the time being, a
continuing social debate on where we want to draw the
A T line in the matter of modern biotechnology is a must.
It is unrealistic to suppose that progress in this field
can be halted, given the speed with which advances
and developments in biotechnology have occurred in
countries like the US, Argentina, Mexico and China.
It’s vital to put this novel technology into practice in a
There is even opposition in the US, where modern sensible and controlled manner to prove safety and
biotechnology has been most widely implemented and usefulness beyond any doubt. To be more specific,
most readily accepted by society. As a result, in May we should now put our knowledge and experience
1998 a coalition of scientists, consumer organisations for instance into the area of high-tech agriculture for
and religious groups filed a lawsuit against the FDA the production of health-promoting foodstuffs and
with the intention of banning 36 genetically modified
11
www.hhs.gov/aspr/barda/bioshield/index.html

Chapter 2: Modern biotechnology: food for discussion! 35

 high-grade products such as medicines by using raise an emotionally charged topic at an early stage
genetically modified (transgenic) plants and animals, was made in Trendanalysis Biotechnology 2007
initially under well-controlled conditions in high- Chances and Choices (a pdf of the full report in
tech glasshouses or stalls. An interesting example Dutch can be downloaded12). The memorandum
is the cultivation of transgenic tobacco plants in looked at ethnicity as a possible factor in scientific
greenhouses for the production of anti-bodies to treat research, genetic diagnostics and genetic population
infections caused by the West Nile virus (Lai et al., studies. The conclusion stated: “The genotype for
2010). When sufficient trust in safety and usefulness certain disorders differs from one ethnic group to
is gained, a wealth of opportunities grows on every another. The efficacy of the drug treatment
bush ready to be explored. for diseases is also affected by the
We, the authors of this book, are afraid patient’s genetic background. However,
that the wrong issues are still being ethnicity is an emotionally

A <<
debated. Take the speed with which charged issue in Europe,

H
ALT
HE
whole genomes are mapped today. especially where genetics

CGA
IET
As a result of that the individual is concerned. At present the

GCG
D INE
human DNA passport is now ethnic background of patients

GAA

IC
MED
a little too close for comfort. and clinical trial subjects is

TTC
Is that what we want? And not automatically registered in

C AT
if it happens, who gets to AGG the Netherlands. The absence
approve it, who is allowed of this information hinders genetic
GTT

to use it? Will it have diagnosis, population studies and

>> C

consequences for our ability scientific research. The government

to get insurance cover? Will it turn must decide whether or not to allow the
our health-care system upside down? registration of ethnicity, and if so, for what
Will it have far-reaching consequences for our purpose and under what conditions.” Well-
criminal code? These are examples of questions distributed information and well-prepared
that we should now be debating publicly so as to public debates are, we believe, indispensable in
lay down the limits of what we want in advance. Not this context.
later, when it becomes nigh on impossible to reverse It is not just the Netherlands that is struggling with
developments in progress. these revolutionary developments. Almost ten
years after the controversial poster issued by the
One admirable first move in the Netherlands to
12
www.cogem.net/ContentFiles/Trendanalyse%202007.pdf

36 Part 1: Introduction
 Source: Getty Images

Dutch League for Animal Protection (Figure 2.2), we 2.3. WHY TRANSGENIC PLANTS?
found the extract and poster below in the October
2003 issue of AgraFood Biotech. It seems that such Plants are genetically modified for four application-
provocative campaigns are needed to wake up the oriented reasons:
general public and get them involved in such far- 1. To improve taste, shelf life and/or dietary value, as
reaching developments. The discussions about in the case of Flavr Savr, and to obtain products
Herman have, in any case, helped result in extensive that prevent disease and disorders and have
legislation in the Netherlands. We will return to this healing properties (novel and functional foods;
later in the chapter. A scientific reconstruction of the nutraceuticals). The science in this area is still
public debate on Herman can be found in the PhD in its infancy, but there are nevertheless huge
thesis, A calf is born, by Elmar Theune (2001). expectations for the long term. Transgenic rice and

Chapter 2: Modern biotechnology: food for discussion! 37

 sorghum are examples of existing products, which idea is to insert an optimum combination of three or
will be discussed in Chapter 8. four resistance genes into a so-called ‘gene cassette’
2. With the aim of making a plant easier and less and to incorporate these into a potato variety. Figure
expensive to cultivate. Disease and pest resistance, 2.3 is a photo of one of their field trials. By producing
protection against cold, drought and/or frost, more of these cassettes with varying combinations
immunity to herbicides, these are the properties that of R-genes, it is possible to make a variety flexible
scientists want to impart to plants. Transgenic soya, and therefore resistant in the long term. It is possible
corn, cotton and canola (rapeseed) are examples of to make variations in space or time, as happens
transgenic crops that are already being produced on naturally in the wild family strains, by initially working
a large scale. The primary aims in this first generation for a few years with one gene cassette while keeping
of transgenic plants are disease and pest resistance the other in reserve, or by using them in combination.
and immunity to herbicides. A newer example is the
transgenic tomato which grows very well in salty soil.
The eminent journal Nature Biotechnology published
an article by Zhang and Blumwald (2001) in which they
describe how they had created genetically modified
tomato plants that not only flourish but produce edible
fruits on extremely salty soils, because the tomato
fruit doesn’t take up salt from the soil. According to
the researchers their discovery means that 60 million
hectares of land around the world will immediately Figure 2.3. Original potato variety Désirée is diseased
become available for the cultivation of such transgenic, and surrounded by healthy Désirée plants supplied with
salt-tolerant crops; land where previously not much a Phytophthora resistance gene (thanks to the DuRPh
would grow. A promising approach to combating researchers14 for the photo).
the notorious potato late blight Phytophthora
infestans comes from Wageningen University13. According to the researchers this method of genetic
By incorporating combinations of resistance genes modification makes it possible to create sustainably
(R-genes) from resistant wild potatoes into existing resistant varieties of much loved classics, such as
commercial potato varieties (cisgenesis), researchers the more than one hundred year old Bintje or Russet
at Wageningen believe that they can protect these Burbank. It is precisely this that makes the approach
potato varieties sustainably against this disease. The so elegant. The researchers are also devising

13
www.cisgenesis.com 14
www.durph.wur.nl/UK

38 Part 1: Introduction
 strategies for this resistance management. The produced commercially on a small scale for use as
question is whether it can also be used in the short chemicals. In recent years there has also been more
term. Genetically modified potatoes for consumption focus on the legislation, particularly concerning the
are still taboo within Europe and so for the moment do manufacture of pharmaceuticals from transgenic
not get further than the glasshouses and trial fields, plants (Spök et al., 2008). Ramessar et al. (2008)
thus still far from our tables. The researchers argue, are looking at these things in detail for transgenic
however, that the cisgenic crops only contain genes corn as a production platform.
that can be used in classical breeding too. They 4. For obtaining plants as alternative feedstocks for
argue that cisgenesis is as safe as or even safer than the production of biofuels. At CHI’s 2009 Advanced
classical breeding and propose a faster and cheaper Biofuels Development Summit there was a
approval of cisgenic crops. consensus that members of the biofuel industry
3. For obtaining plants that produce pharmaceutical are ready to meet the challenge of producing
and other high-value substances. Although plants replacements for petrochemical fuels that will be
have long been the source of many raw materials for cost-competitive and renewable, and that will meet
the pharmaceutical industry (a quarter of medicines the increasingly stringent demands of the Green
contain substances of plant origin), this terrain is Revolution15. Nowadays, we indeed see that there
still virtually barren as regards transgenic plants, are huge investments in the shift from an oil-based
especially compared to transgenic animals. There to a biomass-based society. Today we are also
is, however, enormous potential, partly because experiencing the debate on the question Food or
diseases (such as BSE) that can be transmitted to Fuel?, which we endorse. However, we leave it with
humans are much less of a problem in the production this statement and focus solely on food and health
of plants. Since the mid ‘90s an increasing number in this book. Biofuels are books in themselves (e.g.
of articles have outlined the possibilities of this so- Vertes et al., 2009).
called molecular pharming with transgenic plants The long-term opportunities and possibilities for
and the market is fast approaching. Following small, high-tech countries like the Netherlands are
hundreds of field trials in the United States and the development and testing of new technologies,
Canada dozens of medicines manufactured by especially for items 1, 2 and 4, because in this respect
using transgenic plants have already entered the current ‘farming’ methods will not differ greatly from
various phases of clinical trials, including several those required for transgenic crops. As regards item
in a very advanced phase. Five of them, including 3, some things are a little different. The agricultural
human lactoferrin and lysozyme from rice, are being acreage required for this is small, but the technology

15
www.genengnews.com/gen-articles/alternative-feedstocks-boost-ethanol-production/2942/

Chapter 2: Modern biotechnology: food for discussion! 39

 is advanced. That is where the principle development on the product, herd sizes must be several dozen
opportunities and production possibilities lie for (Factor IX - a drug for haemophilia B - made from
countries like the Netherlands. transgenic sheep or pig’s milk) up to several
thousand (α-anti-trypsin - a drug for cystic fibrosis
PLANTS ARE A GREAT RESOURCE - made from transgenic sheep or goat’s milk). The
FOR THE PHARMACEUTICAL INDUSTRY conventional reproduction methods require a lot of
time and money and there is also a good chance
SI SENHOR, OF COURSE...
...WE’RE PHARMACISTOS! that the foreign gene gets lost in the process.
The requirement for efficient reproductive cloning
techniques with genetically identical offspring is
particularly high in relation to larger herds. The
quest for this has already resulted in Dolly the
sheep (Textbox 2.4), the first cloned mammal. She
was made from an adult udder cell and died on
13 February 2003 from ailments resulting from
premature ageing.
2. To make them more suitable as organ donors
for xenotransplantation (see also Chapter 12).
Xenotransplantation is the transplantation of
animal organs to humans. American scientists
have transplanted insulin-producing cells from
pigs into monkeys with type 1 diabetes. The
2.4. WHY TRANSGENIC ANIMALS? subsequent average survival of these monkeys
was six months, without insulin injections and
Similar in some aspects to plants, animals are with normal blood sugar levels. In the March 2006
genetically modified for the three following application- edition of Nature Medicine plans were announced
oriented reasons: to conduct this research on humans before 2010
1. To render them suitable for the production of high- (Hering et al., 2006). However, a neck-and-neck
purity (human) proteins, in particular medicines. race between xenotransplantation and stem cell
Dozens of medicines made from transgenic therapy threatens to emerge (see also following
animals are currently in various stages of clinical paragraph and Chapters 12 and 14). Political
trials. For commercial applications and depending policies and social acceptance, as well as

40 Part 1: Introduction
 technological advances, will determine who wins Atlantic salmon, causing this genetically modified
the race in the end, or whether both will come in salmon to grow twice as fast.
equal first. Suggestions that it will grow six times the size
3. To obtain “better” farm animals of a normal salmon are pure
and fish. As an illustration of SALMON INDUSTRY THRIVES fantasy: the final size is the
this, work is being done THANKS TO AQUADVANTAGE SALMON same. However, possibly the
on improving disease and most significant advantage
plague resistance, on pigs that for the industry is that the
can digest cellulose, pigs AquAdvantage salmon will
with leaner meat, sheep that also continue to grow in cold
can use low-cysteine feed, conditions. This is in contrast to the
frost-resistant salmon (with non-modified variety and means that
anti-freeze genes from cold- modified salmon can be harvested
resistant fish from the Arctic twice a year instead of the
and Antarctic oceans) and usual once. They also need
giant salmon. An example 30% less food to reach their
of the latter is the AquAdvantage salmon, harvesting weight. AquaBounty Technologies,
made by inserting a gene from an eel and the the producer, expects this salmon to be on the
growth hormone gene of a specific salmon into the market by 201016.

TEXTBOX 2.4. adult cell in this way – an achievement which for years
Dolly the clone. leading scientists deemed biologically impossible.” This
is a literal quote from the inside of the jacket of Gina
“On 5 July 1996 the most famous lamb in history entered Kolata’s book “Clone: the road to Dolly and the path
the world, head and forelegs first. No one broke open ahead”, a very worthwhile read, published in 1998, about
champagne. No one took pictures. Yet the birth of this the history of Dolly and all the events surrounding it.
remarkable creature was soon to provoke amazement
and wonder around the world. Created in Edinburgh’s Dolly was created from a so-called specialised cell,
Roslin Institute by embryologist Ian Wilmut, Dolly was in this instance an udder cell, from a six-year-old
born not from the union of a sperm and an egg, but from sheep. Because DNA undergoes “wear” with every
the genetic material of a single cell taken from a six-year- cell division, Dolly emerged from relatively old genetic
old sheep. Dolly is the first creature to be cloned from an
16
www.aquabounty.com

Chapter 2: Modern biotechnology: food for discussion! 41

 material, immediately giving rise to the question of unspecialised or stem cells (diagram below), which can
how old Dolly actually was at birth. Three years later for example be isolated from embryos, umbilical cord or
it was revealed that Dolly had aged relatively quickly bone marrow. This is one of the reasons why stem cell
and in 2003 she died of early ailments of old age. research is the focus of so much interest (Section 2.5
In theory this problem can be prevented by cloning with and Chapter 13).

Donor of Donor cells

Donor of unfertilised ova Donor sheep Unspecialised
unfertilised ova embryonic cells

In the test tube

be In the test tube
be

Unfertilised Donor udder cell

ovum (specialised) Unfertilised ovum Donor cell

Cell nucleus Cell nucleus Cell nucleus

removed is removed is isolated
The cells are cultured in starvation conditions.
As a result, an adult cell is (possibly) returned to
the unspecialised stage of an embryonic cell Cell nucleus is inserted
into empty ovum
with a micropipette
Ovum without Ovum without
nucleus Cell nucleus
nucleus

The cell nucleus is diploid (2n),

Electrofusion Cultured starved and therefore contains all
by electric shock udder cell chromosomes of the donor
cell in duplicate.

A number of zygotes are

implanted into the uterus
of a surrogate mother
Zygote

Surrogate mother 1
The cloned cell allows for
120-cell multiplication to a 120-cell
embryo embryo in the test tube

When the zygotes have developed into an

embryo of about 120 cells, this embryo is
then inserted into a second surrogate mother

Surrogate mother 2 Dolly Surrogate mother Cloned lamb

The diagrams are adapted versions of the ones originally drawn up by Jos van den Broek, currently Extraordinary Professor in
Biomedical Scientific Communication (Leiden University, the Netherlands).

42 Part 1: Introduction
There are a multitude of long-term opportunities and controversy and about which opinions differ widely.
possibilities in all these areas. The applications in On paper they look like a panacea for almost every
1 and 2 require highly specialised knowledge and disease, but the practical obstacles that still need to
experience, not only of the molecular biology of be negotiated are many and complex and constitute a
animals, but also of industrial high-tech farms, while major challenge for biotechnological researchers. We
only a small amount of land is required. In short, an will briefly look at them here and come back to them in
ideal starting point for countries like the Netherlands more detail in later chapters.
to develop and test new technologies, and to set up
BIOMEDICAL THERAPY HAS LONG SINCE MOVED ON
new production projects with transgenic (cloned) FROM THE EXPERIMENTAL STAGE
animals on high-tech farms. The Dutch company
Pharming, the “creator” of Herman and a global leader
in this area, is a good example of this. However, the
WHOAH! THIS SHOULDN’T
introduction of an animal welfare act, which bans HAPPEN ANYMORE!
nuclear transplantation (see also Chapter 14) - the
cloning technique used by Pharming - together with GRRR
economic and epidemiological reasons, has forced
this company to look abroad, at least as far as
production is concerned. Thus, the Dutch legislation is
now restricting opportunities. This is also a conclusion
in the latest Trendanalysis Biotechnology 2009. In
this Dutch memorandum commissioned by the Dutch
government, one of the biotechnology dilemmas for
policy and politics reads: “Cloning of animals is not
allowed in the Netherlands, but an import ban on
cloned animals, descendants or products of cloned
animals is not sustainable; the government faces the
choice to accept import or to adapt legislation.”
Gene therapy is a treatment for curing congenital
2.5. GENE AND STEM CELL THERAPY disorders. Congenital diseases arise because of
an abnormal or missing gene that causes disease
Gene and stem cell therapy are biomedical symptoms. Gene therapy involves inserting a healthy
developments which have already caused a lot of gene into the cells of the patient. It isn’t always

Chapter 2: Modern biotechnology: food for discussion! 43

 necessary to do this in all cells. That would effectively In 1998 a method was published for the first time to
make gene therapy unviable. Around the world there cultivate human embryonic stem cells in vitro, i.e.
are only a few very specific and rare cases in which outside of the body in test tubes or Petri dishes. This
gene therapy has been at all successful in humans. caused a great deal of excitement in the scientific
Unfortunately there are also cases in which the patient world, with visions of a revolution in health care, but
has died as a result of the treatment. Chapter 11 considerable unrest among religious and bio-ethic
contains a more extensive discussion of the positive groups and public policy-makers. The latter protested
and negative aspects of gene therapy. primarily against the ‘misuse’ of human embryos.
In contrast to gene therapy, in at least one application, However, the real stem cell hype occurred in April
stem cell therapy has long since moved on from the 2001, when the American biologist Daniel Orlic (Orlic
experimental stage. Stem cells from bone marrow have et al., 2001) wrote an article in the leading scientific
been used for more than 30 years in the treatment journal Nature describing how he had largely repaired
of some forms of cancer, for example leukaemia. a mouse heart, damaged following a heart attack, by
Chemotherapy kills not only cancer cells but also most injecting the animal with bone marrow cells. The cells
other growing cells, including the stem cells in bone appeared to develop into heart cells, dramatically
marrow. Which is why bone marrow is taken from the improving the functioning of this organ, but the exact
patient before chemotherapy is started. Where possible, role of the cells in the repair remained unclear.
healthy stem cells are isolated, then stored and injected “The principle of stem cell therapy is as simple as it is
into the patient after the therapy, to provide more healthy fantastical”, said stem cell expert Christine Mummery
growing (blood) cells. If this doesn’t work, bone marrow in the January 2007 issue of a Dutch newspaper
from a donor is used. (NRC). “Stem cells can develop into specialised tissue
Roughly speaking there are two sorts of stem cells: and thus replace diseased tissue. So, new heart
embryonic stem cells and adult stem cells. As the muscle tissue could be created after a heart attack,
name suggests, the former are taken from embryos. or Parkinson’s disease could be cured by creating
When they are isolated from embryos that have only new brain cells. But after years of research we still
undergone a limited number of cell divisions, they have no idea what really happens when stem cells
can grow into any kind of body cell. Adult stem cells are introduced.” In the same year she also stated that
are isolated from bone marrow, umbilical cords, etc., stem cell research was advancing slowly and with
and are already slightly differentiated. They can only varying success mainly because President George
develop into a limited number of related cell types, for Bush had banned the use of government money for
example, blood stem cells into blood cells such as red embryonic stem cell research. She believed that the
and white blood cells. EU was also holding back subsidies for stem cell

44 Part 1: Introduction
research. All this basically means that stem cells have This ministerial decision ended that treatment. Only
far from fulfilled their potential. “Stem cell therapy in university hospitals and the Dutch Cancer Institute
this form is still in a research phase, so treatments were granted a permit to conduct stem cell research.
outside the application of scientific research are Furthermore, permits are only issued if the Central
banned”, said the Dutch Ministry for Health in early Committee on Research involving Human Subjects
2007. In so saying, the minister answered the call to gives approval. This judgement puts the focus firmly
stop stem cell pirates made a year earlier by various on safety, detailed background studies, accurate
medical specialists and stem cell experts. Unlike in research design and comprehensive information to
Spain, Belgium and the United Kingdom, commercial patients - and rightly so. At least in the US, however,
stem cell therapy had been permitted until then in the a fair wind has begun to blow for stem cell research
Netherlands, and for several years already two Dutch since the arrival of Bush’s successor, Barack Obama,
companies had been treating hundreds of incurably in January 2009; more about this in Chapter 14.
sick patients suffering from, for example, a spinal
cord lesion or multiple sclerosis, with expensive, 2.6. EU LEGISLATION
unproven and unauthorised stem cell therapies.
In the EU two provisions concerning legislation on
STEM CELL PIRACY
products involving modern biotechnology came into
force in 2003 and are now being implemented: one
concerning genetically modified foodstuffs and animal
feed, and one concerning the traceability and labelling
of genetically modified organisms and the traceability
of foodstuffs and animal feed produced with genetically
modified organisms. The cornerstone of the EU risk
analysis regarding safety in food and animal feed is
the European Food Safety Authority (EFSA). In close
collaboration with national authorities and in open
consultation with all stakeholders, the EFSA gives
independent advice and makes sure there is clear
communication on existing and expected risks. Figure
2.3 gives an overview of the EU approval procedure
for food and feed originating from genetically modified
plants. The guidelines followed in this process are

Chapter 2: Modern biotechnology: food for discussion! 45

 open to the public and can be downloaded from the conventional equivalent with a history of safe use. This
EFSA website . 17
comparison looks, for example, at the phenotype (the
external appearance of an organism, determined by the
The legislation does, however, differ between EU interaction of the genetic properties – the genotype - and
member states and non-EU countries, but the safety the environment) and chemical composition. Further
assessments in the various national procedures are safety tests are carried out on the basis of perceived
nevertheless based on the internationally recognised differences. The usual items in this risk analysis are
consensus approach of comparative safety analysis the possible presence of toxins and allergens, the risk
(substantial equivalence). This approach involves of transferring the imported genes, and unintentional
comparing the transgenic crop in question with a effects as a result of the genetic modification.

Applicant
application

Member State
transmission
of dossier
EFSA
safety detection
dossier method
Member States Joint Research Center

comments
EFSA
Scientific Panel validation

opinion

European Commission
draft decision
Standing Committee on the
Food Chain and Animal Health
unfavourable favourable
opinion opinion
Council of
European Commission
Ministers

decision

Figure 2.3. Overview of EU approval procedure for GM food and feed (ticks indicate points at which safety is assessed;
thanks to Gijs Kleter for the overview, RIKILT Wageningen, the Netherlands).

17
www.efsa.europa.eu

46 Part 1: Introduction
The second provision concerning traceability in renowned scientific journals. Many of them were
and labelling requires all foodstuffs and foodstuff used in the writing of this chapter. Using this analysis
ingredients originating from genetically modified we have come to the conclusion that there are still
crops, including those without perceivable transgenic multiple opportunities and possibilities for the EU to
material, to be labelled. In addition, there must be a develop into a leading transgenic production area.
documented system of traceability. Labelling is not There is also a firm basis for researching standards
required if the products originate from conventional and norms for the safe implementation of these new
crops, where unavoidable combination with the technologies, without any unacceptable risk to health
transgenic equivalent is less than 0.9 per cent. In some and the environment. A lot depends on future EU-
member states, for example the Netherlands and governments policies in this area as well as social
Germany, GMO-free labelling (i.e. prepared without acceptance. Science and industry are ready. The
gene technology) is permitted. But the criteria here are farmers are generally well-trained, have the necessary
even stricter (Kleter & Kuiper, 2006). experience and are looking for alternatives. In short: it’s
time for the EU to seize the day and become a leading
2.7. CONCLUSION transgenic production continent. In the next chapter we
propose seven points that need attention to remove the
In 2009 we carried out an analysis of the controversies EU hesitation toward gene technology in agriculture.
provoked by modern biotechnology. This analysis is The website GMO Compass18 has been created using
based on a long list of articles that, almost without EU money and is a very good source of information on
exception, were published after the turn of the century new developments in modern biotechnology.

18
www.gmo-compass.org/eng/home

Chapter 2: Modern biotechnology: food for discussion! 47

 2.8. SOURCES Orlic, D., Kajstura, J., Chimenti, S., Jakoniuk, I., Anderson,
S., Li, B., et al. (2001). Bone marrow cells regenerate
Hering, B. J., Wijkstrom, M., Graham, M. L., Harstedt, M., infarcted myocardium. Nature, 410(6829), 701-705.
Aasheim, T. C., Jie, T., et al. (2006). Prolonged diabetes Ramessar, K., Sabalza, M., Capell, T., & Christou, P. (2008).
reversal after intraportal xenotransplantation of wild- Maize plants: an ideal production platform for effective
type porcine islets in immunosuppressed nonhuman and safe molecular pharming. Plant Science, 174(4),
primates. Nature Medicine, 12(3), 301-303. 409-419.
Kleter, G. A., & Kuiper, H. A. (2006). Regulation and risk Spök, A., Twyman, R., Fischer, R., Ma, J., & Sparrow,
assessment of biotech food crops. In P. K. Jaiwal & R. P. (2008). Evolution of a regulatory framework for
P. Singh (Eds.), Metabolic engineering and molecular pharmaceuticals derived from genetically modified
farming II, Plant genetic engineering (Vol. 8, pp. 311- plants. Trends in Biotechnology, 26(9), 506-517.
337). Houston TX, Studium Press. Theune, E. (2001). A calf is born: a reconstruction of the
Lai, H. F., Engle, M., Fuchs, A., Keller, T., Johnson, S., public debate on animal biotechnology. PhD Thesis,
Gorlatov, S., et al. (2010). Monoclonal antibody Wageningen.
produced in plants efficiently treats West Nile virus Vertes, A., Qureshi, N., & Yukawa, H. (2009). Biomass to
infection in mice. Proceedings of the National Academy biofuels: strategies for global industries, Wiley.
of Sciences of the United States of America, 107(6), Zhang, H., & Blumwald, E. (2001). Transgenic salt-tolerant
2419-2424. tomato plants accumulate salt in foliage but not in fruit.
Martineau, B. (2001). First fruit: The creation of the Flavr Savr Nature Biotechnology, 19(8), 765-768.
Tomato and the birth of biotech foods. McGraw-Hill
Companies.

48 Part 1: Introduction
3 GENETICALLY MODIFIED CROPS
AND THE EUROPEAN UNION

Outside the European Union (EU), the area planted with genetically modified crops (GM crops) increases about
10% annually (see Figure 2.1 in preceding chapter). Within the EU there are still seemingly unbridgeable differences
in opinion and acceptance among the Member States. The EU regulation to approve GM crops is very restrictive. By
analysing the controversial issues, especially concerning food and environmental safety, we arrive at seven points
that need attention to remove EU hesitation towards gene technology in agriculture (Table 3.1). We have included
this chapter especially for the policymakers, but we hope that it will interest the layman as well. This chapter is
complementary to Chapter 8.

BRUSSELS IS RUNNING BEHIND ON GM LEGISLATION

NO, MISTER BARROSO. IT’S A LONG WAY

FROM BEING DANGEROUS ...

J. Tramper, Y. Zhu, Modern Biotechnology – Panacea or new Pandora’s box? 49

DOI 10.3920/978-90-8686-725-7_3, © Wageningen Academic Publishers 2011
 3.1. INTRODUCTION To convince all EU members to say yes, many EU
consumers and in particular politicians, must shift
Global food supply is increasingly under pressure as a their attitudes toward GM crops. The first step is to
result of the ever-expanding world population and the recognise the advantages; a step taken for many
growing welfare in developing countries. The transition years already by the Vatican (Textbox 3.1). Convincing
from fossil-based processes to biomass-based examples of GM crops that result in environmental
production, in particular fuels, further aggravates this protection, better land use, phytoremediation (the use
(Boddiger, 2007; Gressel, 2009). In the future, agriculture of (GM)plants to decontaminate polluted water and
will no longer focus solely on the production of food and soils), and especially health benefits for individuals,
feed, but also on fuels, chemicals and pharmaceuticals. will stimulate this shift and step.
This will further increase the pressure on the food
supply. Drought and flood also threaten a sustainable THE VATICAN APPROVES GENETIC MODIFICATION
food supply. To deal with all these challenges, full
input from modern biotechnology, in particular gene
technology is in our opinion an inevitable and essential
measure. However, the issue of whether or not to adopt
genetically modified crops (GM crops) is still a cause of
heated debate in the European Union (EU). By analysing
the controversial issues we come to seven points (Table GM

3.1) that need attention to remove hesitation towards

gene technology in EU agriculture, and to eventually
obtain a firm ‘yes’ to modern biotechnology.

Table 3.1.
Seven focal points to accept GM crops within the EU.

1. Long-term studies to monitor the safety of GM food/crops.

2. Do not polarize!
For the main GM crops, i.e. herbicide-resistant/
3. Global uniformity.
tolerant and insect-resistant crops, positive farm and
4. Risk assessment.
environmental impacts have been proven. Strategies
5. Development of a SMART legislative framework.
for better land-use are now developed by breeding
6. Responsible progress hand in hand with ongoing public
debate. new crop varieties resistant to drought, salinity and

7. Integrated approaches for Third World countries. other extreme environmental stress. Removal of

50 Part 1: Introduction
cadmium from the soil by GM tobacco is a promising global commitment to creating a modern agricultural
example of phytoremediation (Abhilash, Jamil, & infrastructure everywhere and poses the question
Singh, 2009; James & Strand, 2009; Macek et al., whether we have the will and the wisdom to make
2008). The publication on anthocyanin-enriched GM it happen. Miller (2009): “The Golden-Rice story
tomatoes is an example of a health benefit (Butelli makes it clear that the answer is, not yet.” In the
et al., 2008). However, an equally momentous following sections we discuss the two main concerns
achievement of genetic modification of plants, the raised by the opponents of GM crops, i.e. food and
Golden Rice developed a decade ago, has been environmental safety. Our suggested seven attention
largely ignored, because intransigent opposition points (Table 3.1) focus on these two safety issues.
by anti-biotechnology activists makes risk-averse
regulators adopt an over-precautionary approach
that stalls approvals (Miller, 2009). Fedoroff (2008)
argues that a new Green Revolution demands a

TEXTBOX 3.1. Justice and Peace in Rome in November 2004, fellow

Vatican: GM not against God’s will. officials sidestepped taking a definitive stance, insisting
that they were still in the preliminary stage of gathering
This is the title of an announcement in AgraFood information on this difficult issue, while also maintaining
Biotech No 19, December 8, 1999. In October 1999 that this technology “should not be abandoned” (Fox,
members of the Pontifical Academy for Life presented 2004). Five years later Nature Biotechnology headed
two volumes of documents on ethics and genetic a brief announcement with: Vatican cheers GM
technology, the result of more than two years of (Meldolesi, 2009). It announced a closed door meeting
discussion and study. The documents voiced a prudent at the Vatican in May 2009 with leading scientists
yes to GM plants, as the academy was increasingly gathering to discuss a campaign backing agricultural
encouraged that the advantages outweigh the risks. biotechnology. The study week was organised by
Risks should be examined thoroughly, but ‘without a Potrykus, co-inventor of Golden Rice, on behalf of
sense of alarm’. A spokesman of the academy said the Pontifical Academy of Sciences. The Vatican has
they do not agree with groups or persons (e.g. Prince long been concerned about food security, and advisors
Charles), who say GM is against the will of God. Since from the academy, which has a membership list of the
then the Vatican has had an ongoing debate on this most respected names in 20th-century science, have
topic. For instance, by the end of the seminar “GMO: recognised that plant biotechnology has the potential
threat or hope”, convened by the Pontifical Council of to benefit the poor.

Chapter 3: Genetically modified crops and the European Union 51

 3.2. SEVEN FOCAL POINTS TO ACCEPT GM necessary. Dona and Arvanitoyannis (2009) have
CROPS WITHIN THE EU analysed possible health risks of GM crops. They write
that intensive work is currently in progress to thoroughly
1. Long-term studies to monitor the safety of GM understand and forecast possible consequences on
food/crops human, animals, and the environment, but anticipate
More than a decade of consuming GM foods has that many years of careful, independent research are
demonstrated that they have no direct health risks and needed to accomplish this. They believe that these
that the required safety assessments have worked long-term studies are especially needed to take away
well. Domingo (2007) concludes though, that relatively any doubt that may exist in society. Although we do
little scientific toxicological research has been done not support the whole paper, we endorse this last
on GM crops/foods and that long-term studies are view. The first item requiring attention is thus, in our

TEXTBOX 3.2. benefits, and not just the risks, in its deliberations
The 20-year environmental safety record of GM trees. of field trials and releases. Their search in publicly
accessible databases worldwide revealed >700 field
In a commentary of May 2009 in Nature Biotechnology trials with GM trees (including forest trees, fruit trees
Strauss et al. (2009) call for more science- and woody perennials). None of them has reported
based (case-by-case) evaluation of the value and any substantive harm to biodiversity, human health
environmental safety of GM trees, which requires or the environment. Few GM tree species have as
field trials. However, the regulatory impediments yet been deployed commercially. A notable exception
being erected by governments around the world are is Bacillus thuringiensis toxin(Bt)-expressing poplar
making such testing so costly and Byzantine, it is trees in China. Approximately 1.4 million Bt poplars
now almost impossible to undertake field trials on GM have been planted in China on an area of ~ 300 –
trees in most countries. One year later, in a letter to 500 hectares along with conventionally bred varieties
the editor of the same journal, Walter et al. (2010) to provide refugia to avoid the development of Bt
summarise the key published evidence relating to the resistance in insects. The trees are grown in an area
main environmental concerns surrounding the release where economic deployment of poplar was previously
of GM trees. On the basis of their analysis of a very impossible due to high insect pressure. GM trees have
large amount of performance and safety data related been successfully established and have successfully
to GM crops and trees gathered since field trials were resisted insect attack. The oldest trees in the field are
first initiated in 1988, they pled for a consideration of now 15 years old. No harm to the environment has
the opportunity costs for environmental and social been reported.

52 Part 1: Introduction
opinion, a clear definition of how long-term studies companies approached the chief policymakers in
should be conducted, and a proposal that these should President Reagan’s administration (1981-1988)
apply to any novel crop/food product. The latter is in with a request to set up more restrictive regulations
line with what Kok et al. (Kok, Keijer, Kleter, & Kuiper, concerning the acceptance of GM crops (Miller et
2008) propose, i.e. to develop a general screening al., 1997). Their suggestions went considerably
frame for all newly developed plant varieties to select further than could be justified by scientific reasoning,
varieties that cannot, on the basis of scientific criteria, but their motives were clear: regulations as market
be considered as safe as plant varieties that are barriers for less powerful competitors such as seed
already on the market. They conclude that the current companies and biotech starters. Their success has
process of the safety evaluation of GM crops versus led to the present overregulation, strongly limiting the
conventionally bred plants is not well balanced. And introduction of new GM crops, but ironically hitting
we fully support this view. An interesting case is the the multinationals themselves most. Ten years later
safety record of GM trees (Textbox 3.2). Miller (2008) fulminated again and condemned the
decision taken by two university rectors in Germany
2. Do not polarize! to forbid scientists to continue their field trials with GM
Scientifically there is indeed no reason to test GM crops. He accused German universities of protecting
foods more thoroughly than other new food products. their reputations by curtailing the academic freedom
The fact that it happens, is not only the result of of faculty and students in the face of demands and
campaigns by anti-biotechnology organisations. In threats from ideological bigots. In another paper Miller
the early 1980s, under the pretext of environmental and co-authors (Miller, Morandini, & Ammann, 2008)
protection, some of the largest agricultural chemical take a sharp stance against the publication policy

TEXTBOX 3.3. But pseudo controversy and sensational claims have

Modern biotechnology: scientific victim? originated within the scientific community as well, and
even scholarly journals’ treatment of the subject has
“Primarily outside the scientific community, been at times unscientific, one-sided and irresponsible.
misapprehensions and misinformation about These shortcomings have helped to perpetuate ‘The
recombinant DNA-modified (also known as ‘genetically Big Lie’ that recombinant DNA technology applied
modified’, or ‘GM’) plants have generated significant to agriculture and food production is unproven,
‘pseudo-controversy’ over their safety that has resulted unsafe, untested, unregulated and unwanted. Those
in unscientific and excessive regulation (with attendant misconceptions, in turn, have given rise to unwarranted
inflated development costs) and disappointing progress. opposition and tortuous, distorted public policy.”

Chapter 3: Genetically modified crops and the European Union 53

 of some leading scientific journals. By publishing Health Organization, require an estimate of any
activists’ papers with sensational, inaccurate claims unpredictable and unintended effects, even if there
they provoke, according to Miller et al. (2008), pseudo- is no indication that such effects are more likely to
controversy, misapprehensions and misinformation occur in GM crops than in conventional ones (Batista
about GM crops, especially concerning environmental & Oliveira, 2009). There are two different approaches
or health risks (Textbox 3.3). for this purpose (Kuiper, Kok, & Engel, 2003). One is
a targeted approach that is regularly used to evaluate
We think that Miller and co-authors are right in new GM foods. Here, several key nutrients are
principle, but the way they express it will not unite the analysed that, if inadvertently altered, could influence
warring parties, and they are not the only ones using the nutritional value and eventually the safety of the
forceful language. We believe that, for the time being, modified product. This approach does not consider any
too many tests may be preferable to too few, given the unknown anti-nutrients and natural toxins. The second
ever-present hypersensitivity to GM food and crops approach is non-targeted and based on profiling
among the opposition groups, but also among many methods, in which potential alterations in GM food that
policymakers and consumers. occur at the genomic level, as well as at the levels of
gene expression, translation and metabolic pathways,
3. Global uniformity are evaluated. Several recent studies have begun
Some opposition groups request that lengthy to explore profiling methods that aim to increase the
toxicological tests lasting at least two years should probability of detecting any unpredictable, unintended
be conducted with GM foods. The European Food effects and, consequently, improve the efficiency
Safety Authority (EFSA) reports that 90-day food of GM food safety assessment (Batista & Oliveira,
tests on animals, mainly rats, are usually sufficient 2009). Profiling techniques are a potentially powerful,
to demonstrate the safety of GM foods, provided that complementary tool, offering the capacity to broadly
these tests are performed according to international screen for possible changes at different integration
guidelines (Konig et al., 2004). The report calls for a levels of cells or tissues in a non-selective, impartial
more uniform approach to food testing and the use manner. For these reasons we have formulated this
of new (profiling) technologies. It suggests a solid third attention point.
pre-market risk analysis rather than monitoring after
a product has been marketed. Risk assessment of 4. Risk assessment
predictable effects is easily attained through specific Environmental safety policy is generally built on the
in vitro and clinical tests. Some institutions, such as precautionary principle (Textbox 3.4). The precautionary
the Food and Agriculture Organization and the World principle, for example, is the basis of EU Directive

54 Part 1: Introduction
2001/18, which states: “In accordance with the According to them this indicates that arable crops are
precautionary principle, the objective of this Directive is unlikely to survive for long outside cultivations, but it
to approximate the laws, regulations and administrative does not mean that other genetic modification could
provisions of the Member States and to protect human not increase weediness or invasiveness of crop plants.
health and the environment…” For GM crops it can be They conclude: “The ecological impact of plants with GM
assumed that pollen from a GM crop will pollinate a wild traits such as drought tolerance or pest resistance that
variety, if they grow in each other’s neighbourhood. GM might be expected to enhance performance under field
seeds spread with wind or birds, and end up in the wild. conditions will need to be assessed experimentally when
It is thus conceivable that the (pollinated wild variety such plants are developed.” The same holds for GM
of) GM crops in the natural environment could become plants for phytoremediation (Gressel & Al-Ahmad, 2005)
agriculturally problematic. This can be prevented and GM plants for production of pharmaceuticals. GM
by thorough testing, first in the lab, then in a closed crops usually differ from their conventional counterparts
greenhouse followed by contained field testing and finally only with respect to one or a few desirable genes, in
an extensive period of monitoring once the GM crops are contrast to crops from traditional breeding methods
being cultivated on a large scale. Except for the latter, that mix thousands of genes (Atherton, 2002). Armed
this is largely what is required by many governments. with genomic information and nanotechnology, plant
Concerning GM crops, the risk of rampant growth in molecular biologists are redesigning molecular toolkits
the natural environment is very small, and most of them to engineer plants still more precisely (Moeller & Wang,
will not survive anyway. For example, rape-seed plants, 2008). It would thus seem logical that GM crops pose
both transgenic and conventional, were cultivated in a fewer risks than conventionally modified crops.
field and studied by researchers for 10 years without Substantial scientific data on the environmental effects
harvesting. After 5 years there was not a single GM of the currently commercialised GM crops are available.
plant to be seen, and after 10 years there were just a Sanvido et al. (2007) have reviewed this scientific
few conventional ones. Crawley et al. (Crawley, Brown, knowledge derived from the first 10 years of worldwide
Hails, Kohn, & Rees, 2001) state: “Four different crops experimental field research and commercial cultivation.
(oilseed rape, potato, maize and sugar beet) were grown The review focuses on the currently commercially
in 12 different habitats and monitored over a period of 10 available GM crops that could be relevant for agriculture
years. In no case were the genetically modified plants in Western and Central Europe (i.e. maize, rapeseed
found to be more invasive or more persistent than their and soybean) and on the two main GM traits that are
conventional counterparts.” Their results concern GM currently commercialised, i.e. herbicide tolerance and
traits (resistance to herbicides or insects) that were not insect resistance. The sources of information include
expected to increase plant fitness in natural habitats. peer-reviewed scientific journals, scientific books,

Chapter 3: Genetically modified crops and the European Union 55

 reports from regions with extensive GM-crop cultivation, crop cultivation on the environment by considering
as well as governmental reports. The data available so the impacts caused by cultivation practices of
far provide no scientific evidence that the cultivation of modern agricultural systems. Even without GM crops,
the presently commercialised GM crops has caused modern agricultural systems have profound impacts
environmental harm. The authors recognise, though, on all environmental resources, including negative
that results from large-scale cultivation systems, impacts on biodiversity. When discussing the risks
which GM crops generally are, have to be transferred of GM crops, one has thus to recognise that the real
with care to small-scale agricultural systems like in choice for farmers and consumers is not between
Switzerland. The interpretation of results is often GM technology that may have risks and a completely
challenged by the absence of a baseline for the safe alternative. The real choice is between GM crops
comparison of environmental effects of GM crops in and current conventional practices for pest and weed
the context of modern agricultural systems (Sanvido management, all possibly having positive and negative
et al., 2007). There is thus a need to develop scientific outcomes. To ensure a true precautionary policy, one
criteria for the evaluation of the effects of GM crops on should compare the risk of adopting a technology with
the environment to assist the regulatory authorities. In the risk of not adopting it. This all led us to this fourth
their study, Sanvido et al. discuss the effects of GM- item requiring attention.

TEXTBOX 3.4. that precautionary measures must also be met if there

The precautionary principle. is insufficient scientific proof of harm, but inaction
may lead to irreversible damage or risk for health and
In November 2005 an interesting report on this subject environment. Conversely, the UN Millennium Task
was published by the Institute of Advanced Studies, Force on Science, Technology and Innovation states
United Nations University . The report is called
19
in its report of 2005 that the focus on technological
“Trading Precaution: The Precautionary Principle risks may overshadow the possible benefits of an
and the WTO”. In the introduction it is stated that the up-and-coming technology, because these are often
precautionary principle is central to environmental difficult to predict. Underlying the continuing debate
policy and a key element in multilateral environmental on the precautionary principle is the fundamental
treaties. As such it is a fundamental part of the question of how policy concerning health, safety
Cartagena Protocol20 on Biosafety. Policy makers and and environment should be developed if on the one
officials who use the precautionary principle and are hand there is a lack of scientific consensus and on
involved in environmental and health matters assume the other a significant portion of the population has
19
www.ias.unu.edu irrational (from a scientific perspective) opinions
20
bch.cbd.int/protocol

56 Part 1: Introduction
(fears) about (for) the material concerned. The “In order to protect the environment, the precautionary
precautionary principle endeavours to bridge the approach shall be widely applied by States according
gap between scientific uncertainty and regulation to their capabilities. Where there are threats of
of risk. Circumstances determine the way in which serious or irreversible damage, lack of full scientific
precautions are to be taken. These considerations certainty shall not be used as a reason for postponing
make it difficult to draw up a generally applicable cost-effective measures to prevent environmental
definition of the precautionary principle. International degradation.”
lawyers writing about the precautionary principle
usually start from two ostensibly similar definitions. The major difference is in the word ‘cost-effective’,
The first comes from the Bergen Ministerial linking the need to take measures with the possible
Declaration on Sustainable Development of 1990: economic effect. The debate on the precautionary
principle is complex and often abstract. In a certain
“In order to achieve sustainable development, sense the precautionary principle can be seen as a
policies must be based on the precautionary principle. “rather shambolic concept … muddled in policy advice
Environmental measures must anticipate, prevent, and subject to whims of international diplomacy and
and attack the causes of environmental degradation. the unpredictable public mood over the true cost of
Where there are threats of serious or irreversible sustainable living” (O’Riordan & Cameron, 1994). In any
damage, lack of full scientific certainty should not be case, the result was various different pieces of legislation
used as a reason for postponing measures to prevent between the EU and the US. As such the EU has very
environmental degradation.” strict rules on authorisation and marketing of genetically
modified organisms and products compared to the US.
The second oft-quoted definition is to be found in Conversely, some food products such as unpasteurised
Principle 15 of the Rio Declaration on Environment and cheese are heavily regulated in the US for health
Development, of 1992: reasons, while they are highly valued in the EU.

5. Development of a SMART legislative framework where the number of field trials even fell and only
Despite the ongoing controversies, in 2009 the area a handful of the 27 EU countries cultivated the only
planted with GM crops grew still further. More than 13 GM crop approved there (Bt maize). In this context
million farmers in 25 countries planted GM crops, over Richmond (2008) reviews the precautionary principle
90% of them in developing nations (Marshall, 2009). and believes that the progress made in every area of
This rapid progress is causing more anxiety about biotechnology quickly leads to countless applications
the effect on the environment, especially in Europe, and products to benefit the society. Progress is so

Chapter 3: Genetically modified crops and the European Union 57

 rapid that policymakers, legislative authorities and law will be required as a framework for decision making
enforcers cannot keep up. This harbours the risk of (Textbox 3.5). In line with this we propose the SMART
serious and irreversible environmental consequences approach as the fifth item needing attention.
that will be difficult to control. The challenge is to
develop a legislative framework with effective checks The SMART tool for defining goals21 is well-known in
and balances that help avoid serious and possibly human resource management. Analogously, it must
irreversible consequences but, at the same time, do be possible to base the legislation concerning agro-
not restrict innovation. The precautionary principle biotechnology on its own version of the SMART
demands scientifically acceptable evidence that principle:
no damage will be done, if products are introduced Sustainable–Measurable–Acceptable–Reasonable–
or activities implemented. Determining scientific Time-based
standards and norms (for example, less than 5% Many of the elements of the SMART approach
chance of damaging effects) by (i) asking the already exist in the EU 2000 policy on the application
right questions and (ii) producing an acceptable of the precautionary principle. The EU Commission
experimental design, will lead to an approach that can issued a policy communiqué in 2000 outlining “the
reduce risk and provide policymakers and the public Commission’s approach to using the precautionary
with a better understanding of possible problems in the principle” and to “establish Commission guidelines for
future. Richmond also believes that, because there is a applying it” - Communication from the Commission on
need to better understand and evaluate risks, statistics the Precautionary Principle COM 1. These guidelines

TEXTBOX 3.5. safe until proven otherwise. If the gun was empty, but
Statistics as a framework for decision making. I have accepted the incorrect hypothesis that it was
loaded, I am guilty of statistical error Type II. If the gun
There are two overall categories of statistical errors: was indeed loaded and somebody suggested that it
the rejection of a correct hypothesis (Type I) and the wasn’t, that person was guilty of a Type I error, and, if
acceptance of an incorrect hypothesis (Type II). For the trigger is pulled, may also be guilty of murder. If we
example, there is a gun on the table and there is no now replace the gun with open cultivation of transgenic
information available to establish whether or not it is crops, the doom scenario is clear. The lack of sufficient
loaded. The precautionary principle dictates that it data to show that something is harmful doesn’t mean
must be assumed that all guns are loaded unless the that it is safe; the correct conclusion is that there are
opposite is proven. The alternative approach (often used insufficient data to make a judgement.
for environmental considerations) is that everything is
21
www.topachievement.com/smart.html

58 Part 1: Introduction
clearly state that the precautionary principle should A problem hampering this development of the SMART
be applied in a proportional, non-discriminatory and approach, is the difference between the regulatory
consistent manner, with an examination of the benefits structures underlying US and EU policies regarding GM
and costs of action (or lack of action) and with an foods/crops. The US regulates GM foods/crops more
examination of scientific developments. It is interesting as end products, applying roughly the same regulatory
to note that the EU has failed to live up to its own framework as to conventional ones. The EU, contrarily,
policy. A good starting point for the development of a regulates products of agro-biotechnology more as the
globally uniform, SMART-based legislative framework result of a specific production process. Accordingly,
is the critical and thorough review by Chandler and EU regulates GM foods/crops specifically. As a result,
Dunwell (2008) of hundreds of scientific papers on the pertinent US regulation is relatively permissive,
gene flow, risk assessment and environmental release whereas EU regulation is relatively restrictive.
of GM plants. A good model to start working with is Both Ramjoué (2007) and Hammitt et al. (Hammitt,
wheat (Peterson & Shama, 2005). Wheat varieties Wiener, Swedlow, Kall, & Zhou, 2005) analyse why
produced with modern biotechnologies, such as GM food policies in the US and the EU are different.
genetic engineering and mutagenic techniques, have The fact is that the public debate in Europe has ground
lagged behind other crop species and have only to a halt, having been reduced to a hopeless tug-
emerged recently. This offers a unique opportunity of-war about GM foods/crops. A poignant example
to assess comparatively the potential environmental is the overwhelming majority voting in early 2009
risks (human health, ecological, and livestock risks) against the proposals to overturn national bans on
associated with genetically engineered, mutagenic, GM-maize cultivation in France, Greece, Austria and
and conventional wheat production systems. Hungary (Abbott, 2009). This EU impasse over agro-

EU MEMBERS IN TRENCH WARFARE OVER AGRO-BIOTECHNOLOGY

Chapter 3: Genetically modified crops and the European Union 59

 biotechnology was deepened even further in April 2009 in particular, where they have become strong both in
with the ban on GM maize by the German government. politics and communication. Furthermore, they have
In September 2009 Commission President José much more knowledge of these complex matters than
Manuel Barroso started an initiative to develop rules they generally demonstrate in public. That is a hopeful
allowing member states to ban the cultivation of EU- sign, as is the fact that public opposition seems to
approved crops. Proposals are due mid-2010. Despite be falling away. In the Eurobarometer public opinion
of all this hassle, we as authors still strongly belief in survey of 2008, the percentage of those who said they
employing agro-biotechnology and we challenge our were against GM crops fell from 70 to 58% (Abbott,
colleagues to facilitate responsible progress and to 2009).
inform the public objectively.
SEED COMPANIES HAVE THE POWER TO CONQUER THE WORLD
6. Responsible progress hand in hand
with ongoing public debate. I’M BOND,
The advocates and opponents of modern GENES BOND IT’S OURS,
biotechnology need to stop fighting and start agreeing
ALL OURS!!!
on SMART goals for the future. This means specifically CEO
MONSANTO
and quantitatively defining which risks (if any) are
acceptable, what is meant by “substantially equivalent”
in the principle of substantial equivalence, a heavily
criticised principle22, what is of consequence in genetic
modification of crops, what is sustainable, and what is
natural or organic. To achieve this, the advocates and
more especially the experts really need to understand
that the public has both justifiable and imaginary
concerns; this must be respected. In addition, the
opponents should accept that GM plants are here
to stay and can even offer huge benefits if we deal 7. Integrated approaches for Third-World countries.
with these new technologies skilfully and carefully. The domination of global agriculture by a handful of
Subsidised activist organisations such as Greenpeace multinationals can have adverse effects, especially on
are necessary as a counterbalance in a technological small farmers in the Third World. Anti-trust laws should
society such as the EU in general and the Netherlands prevent this happening, but companies like Monsanto
with the patents on the GM plants, have great power
22
www.i-sis.org.uk/subst.php

60 Part 1: Introduction
and could in a doom scenario take over the world in of experience with scientific decision making that makes
their grip via the food supply chain. During the last 30 Africa hesitant; some of the fears of the new science
years, we have seen that Third-World farmers are able have their roots in mistakes in the past. Europeans,
to adopt new, more efficient technologies and really use for instance, introduced water hyacinth and Nile perch
them. However, it is still true that due to gene technology, in Africa with devastating consequences. So how can
agriculture has become even more dependent on a Africans be sure that GM foods/crops will not lead to
smaller number of large companies. We therefore feel even bigger mistakes? African governments can take
justified in asking whether it is desirable for the situation a number of measures to prevent this, for instance
to continue in this way. The first issue to address then is by building a critical mass of people with the ability to
the plausibility of the claim that GM technology has the evaluate and manage technology within the individual
potential to provide the hungry with sufficient food for countries themselves. A strong scientific community will
subsistence. Carter (2007) discusses this claim within help select the best and most useful biotech applications
the domain of moral philosophy to determine whether and avoid any for which the risks outweigh the benefits.
there exists a moral obligation to pursue this end if In the southern part of Africa alone where current food
and only if the technology proves to be relatively safe production is under the threat of climate change (Lobell
and effective. By using Peter Singer’s duty of moral et al., 2008), around 4 million people depend for their
rescue, she argues that we have a moral duty to assist existence on food donations (Botha & Viljoen, 2008).
the Third World through the distribution of GM plants. Knowing this, it makes sense to consider GM food/crops
She concludes her paper by demonstrating that her as a means of reducing hunger and improving food
argument can be supported by applying a version of quality. Africa did not profit from the Green Revolution
the precautionary principle on the grounds that doing that took place in the West in the middle of the last
nothing might be worse for the current situation. Asante century. The expectation is that gene modification
(2008) criticises opinions and perceptions blocking GM of traditional African food crops such as sorghum will
technologies that can potentially improve survival and produce a second green revolution from which they will
quality of life for millions of people in Africa. We endorse benefit. The entire subject of GM organisms/technology
his view that scientists must help provide an answer is however saddled with different opinions, considerable
to this problem by ensuring that debate on GM crops frustrations, and growing ethical and environmental
addresses facts, not opinions. The initial refusal of badly concerns, globally, leading to the already mentioned
needed food by some African countries in 2002 makes problem addressed by Asante (2008). Scientists in
clear that most of them simply do not as yet have the the individual African countries, and more particularly
experience and scientific capacity to make informed scientists from the West, must help to ensure that
decisions about GM food. However, it is not only a lack debates on GM crops address facts, not opinions.

Chapter 3: Genetically modified crops and the European Union 61

 that it is doubtful whether the development costs of this
SUPERDENSE SORGHUM...
GM sorghum can be justified when compared with the
OVERLOADED WITH VITAMINS!
costs of investing in sustainable African agriculture.
According to them, GM sorghum can only be
successfully introduced if it forms part of an integrated
approach. The Alliance for Green Revolution in Africa
shares this vision. In less than two years, according
to Kofi Annan, this organisation has collected $330
million for a comprehensive and integrated project,
3KG initially in the following six areas23:
2KG

1KG 1. Development of higher yielding, disease-resistant

and climate-resilient varieties of African crops.
2. Seed-multiplication and distribution systems.
3. Improved soil health.
After maize, wheat, rice and barley, sorghum is the
4. Agricultural education.
most important grain in the world and the second most
5. Agro-dealer networks that get inputs to farmers in
important crop on the African continent. In 2006 the Bill
remote locations.
and Melinda Gates Foundation donated $450 million
6. Development of policies that benefit small-hold
to the African Biotechnology Sorghum consortium that
farmers.
consists of companies, institutes and universities in
Southern Africa and North America (Botha & Viljoen,
Issues that include water use, food storage and
2008). It aims at using gene technology to improve the
processing, and market development are also
health and welfare of people in the poorest countries
considered.
of the world by making GM sorghum that is more
As in the rest of the world, examples of GM food that
nutritional and more digestible. The target is a GM
are beneficial for the health of individual consumers
sorghum that contains more essential amino acids,
are badly needed in Africa and other Third-World
especially lysine, but also increased levels of vitamin
countries. Biofortified sorghum is a good start. Naqvi et
A and E, and more absorbable forms of iron and zinc.
al. (2009) reported recently on orange maize with extra
vitamins. Using gene technology German and Spanish
Botha and Viljoen (2008) have carefully analysed all
researchers have enriched South African white maize
the advantages and disadvantages, making use of the
experience gained with Golden Rice. They conclude 23
www.nrc.nl/redactie/binnenland/speeches/kofi_annan.pdf

62 Part 1: Introduction
with beta carotene (a precursor of vitamin A causing the elections, the new Commission President José
the orange colour of the maize), and precursors of Manuel Barroso started an initiative to indeed develop
vitamin C and folic acid. Natural white maize, which is rules allowing the separate member states to ban the
a staple food in many developing countries, contains cultivation of EU approved crops. The appointment of
relatively few vitamins. Compared to the normal maize, John Dalli as Commissioner for Health and Consumer
this GM maize contains as precursor equivalents six policy clearly showed a shift from an anti-to pro-GM crop
times as much vitamin C, twice as much folic acid, policy. Less than a month in office he had already taken
and 169 times as much vitamin A. This means that the most controversial decision a euro-commissioner
consumption of 100 to 200 grams of the GM maize can take: at the beginning of 2010 he approved the
yields the daily recommended amount of vitamin A and cultivation of a second GM crop, i.e. the Amflora potato
folic acid and 20% of that of vitamin C. of BASF. For twelve years all decisions on approvals
were halted. In mid-July 2010, at the time of finishing
3.3. CONCLUSIONS this chapter, he came with a new law proposal giving
the separate countries authority to ban GM crops.
In April 2009 a Dutch proposal concerning whether or According to experts, this proposal creates political
not the decision to cultivate GM crops should be left to room to approve GM crops faster at the EU level. It
individual Member States (Anonymous, 2009), was put gives us the feeling anyway, seeing this all happen,
forward to the EU Council. The then Czech presidency that the Member States are moving slowly towards a
said that a surprising number of countries reacted consensus on lifting the bans, which is indispensable
positively to it. The proposal suggested that a possible for responsible progress at least in some of the Member
solution to GM crops approval issues would be to apply States. The point at which a firm “yes” will be obtained
internal market rules for the import of products – with from all members still seems a long way off, but we
a decision on the EU level, but for cultivation it could believe that it is still not too late, if we pay sufficient
be left to each Member State. In September 2009, after attention to the seven points elaborated in this chapter.

Chapter 3: Genetically modified crops and the European Union 63

 3.4. SOURCES assessment and the environmental release of
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66 Part 1: Introduction
part two
Our daily food and drink
 OUR DAILY FOOD AND DRINK

“There has never been any suggestion that genetically manipulated food is harmful to the consumer. And yet
there are still serious concerns about it. Europe now needs to determine whether the truth is closer to the gloomy
pronouncements of Greenpeace or the risk-free Teletubby-like utopia that the biotech industry presents.”

Rik Nijland, science writer, April 1999

Modern biotechnology is clearly a very hot topic in this present day and age, particularly where our daily food and
drink are concerned. Traditional biotechnology has played an important role in our food production for centuries, but
modern biotechnology has now become an unavoidable part of this process. However, the heated discussions are
chiefly concerned with food from transgenic crops, the so-called gene food, variously called Franken(stein) Food or
monster food by its opponents. In part two of the book gene food is dealt with separately in Chapter 8, as are the
traditional biotechnological products like cheese, bread, wine and meat. For here too, modern biotechnology now
plays a key role.

BIOTECHNOLOGY: A TELETUBBY-LIKE UTOPIA?

OH NO!

69
4 CHEESE:
BIOTECHNOLOGY THROUGH THE AGES

“As their highnesses travelled”, wrote Horace Walpole in an 18th century letter to a friend, commenting on a fairy tale he
had been reading, “they were always making discoveries, by accidents or sagacity, of things they were not in quest of.”

It was Walpole who suggested that the word “serendipity” be included in our vocabulary after reading the Three
Princes of Serendip. Serendip is the old Persian name for Sri Lanka. Nowadays serendipity is defined as the finding of
something unexpected and useful particularly whilst looking for something entirely unrelated, or to use the visual words
of Pek van Andel, studying serendipity and a winner of the Ig Nobel prize: “looking for a needle in a haystack and rolling
out of it with a milkmaid.” Since 1994, Serendip has also been an interactive educational website that helps people
improve their chances of deliberately making discoveries by chance24. The discovery of cheese is a notable example
of serendipity.

SERENDIPITY: LOOKING FOR A NEEDLE IN A HAYSTACK

AND ROLLING OUT WITH A MILKMAID.
WHO CARES ABOUT THE NEEDLE!

24
serendip.brynmawr.edu/serendip

J. Tramper, Y. Zhu, Modern Biotechnology – Panacea or new Pandora’s box? 71

DOI 10.3920/978-90-8686-725-7_4, © Wageningen Academic Publishers 2011
 4.1. OLD CHEESE slaughtered young calf and pouring milk into it, the
same process can be observed: the casein in the
Biotechnology is at least as old as documented milk curdles. Here too a similar kind of enzyme is
history. Before 700 BC Homer, the author of The Iliad responsible for this action, namely chymosin (also
and The Odyssey, the oldest preserved examples of called rennin), which leaks from the stomach wall and
Greek literature, described a simple, yet interesting enters the milk. The chymosin then divides up the
biotechnological experiment. He wrote that if you casein into a large part (90%) that separates out and a
crush a fig branch and then stir the crushed part into small part (10%) that remains dissolved in the residual
milk, a solid forms in the milk, leaving a fluid which can liquid (whey). This must have been a mysterious but
then be drained off. What he was describing here is useful occurrence for observers in ancient times.
the making of a type of cottage cheese. What Homer As far as we’re concerned it is one of the first ever
didn’t and couldn’t know, is that the crushed fig branch biotechnological applications.
oozed a little sap which contained the enzyme ficain
(or ficin). This enzyme causes the casein (curds), the 4.2. TRADITIONAL CURDLING
components in milk that help form cheese, to separate
for the most part, thus making the casein curdle. In the nineteenth century there emerged a little
understanding of what curdling actually involved.
MAKING CHEESE BY USING CRUSHED Furthermore, the first curdling company was founded
FIG BRANCHES IS A VERY OLD TECHNIQUE....
in that century, in 1875 in Copenhagen by a man
EVE, ARE YOU MAKING CHEESE AGAIN?! called Christian Hansen. Hansen bought rennet
stomachs from freshly slaughtered young calves and,
using salt solution, extracted the chymosin from them.
The extract, rennet, is one of the first standardised,
industrial products to be used in a biotechnological
process, i.e. cheese-making. The Christian Hansen
company is still producing rennet today, in virtually the
same way. However, since 2002, the company has
been working on new developments in collaboration
with Novozymes, also a Danish company and
one of the world’s biggest enzyme manufacturers,
which makes frequent use of modern biotechnology
From an even earlier age comes yet another cheese
(Textbox 4.1).
story. By removing the fourth stomach of a freshly

72 Part 2: Our daily food and drink

TEXTBOX 4.1. milk as a pre-treatment process to optimise coagulation
Cheese alliance. 25
and give a higher yield of cheese. The yield increase
is in the order of two percent. That may seem trifling to
On 26 August 2002 Novozymes announced an alliance the layman, but cheese professionals regard it as one
with the international food ingredients company Chr. of the greatest innovations for several decades, given
Hansen, with the initial aim of boosting yields in cheese that all other attempts from the whole dairy industry in
production. The first fruit of this collaboration was launched the previous ten years have only delivered a one percent
in 2005. The product in question was a phospholipase increase in yield. The enzyme works especially well in
(hydrolysing enzyme) which was brought onto the market cheeses like mozzarella (BioTimes December 2005;
under the name of YieldMAX PL. The enzyme is added to newsletter published by Novozymes).

For every 10,000 litres of milk, approximately 1 litre of been conducted. Ironically, Dutch cheese-makers are
rennet is used in cheese-making. That may not sound among the few in the industry who still don’t use this
like much, but considering that in the Netherlands technique (find out why in the next section). This is
alone 700,000 tonnes of cheese are produced each even more remarkable because many of the enzymes
year, starting with 7,000,000,000 litres of milk, for used in food production are currently made by using
which 700,000 litres of rennet are needed (approx. recombinant microorganisms (Olempska-Beer,
1 litre of rennet per tonne of cheese), then it soon Merker, Ditto, & DiNovi, 2006). We will come back to
becomes clear just how many calves’ stomachs are them in the next chapter about bread.
required. Rennet is therefore a scarce and expensive
commodity and the industry has long been anxiously 4.3. MODERN CURDLING
searching for an alternative. With the single exception
of the enzyme from the microorganism Mucor miehei, In the early 1980’s the Dutch biotechnology company,
attempts to bring microbial rennet onto the market had Gist-Brocades (now part of DSM), began experimenting
all been fairly unsuccessful. Until twenty years ago in with recombinant DNA technology. Researchers at
1989, that is, when a Dutch company (the present- Gist-Brocades bought from Unilever the chymosin gene
day DSM-Gist), brought an alternative rennet onto the of a cow; this gene is the piece of DNA that ensures
market which was in quality terms at least as good as the production of the enzyme chymosin in suckling
the natural version. The basis of the technique used calves. They then “inserted” this piece into the genetic
to make this new product was laid down in 1973, once material of yeast cells from Kluyveromyces lactis,
the first successful recombinant DNA experiments had one of their so-called “plugbugs” (Textbox 4.2). These

25
www.novozymes.com/NR/exeres/11CACD69-CDAE-4959-94F9-CECD11D83C66.htm

Chapter 4: Cheese: Biotechnology through the ages 73

 plugbugs then made calf chymosin. The daughter cells Environment declared back in 1994 that he was in
of these yeast cells, which can be cultivated in great favour of the use of the recombinant enzyme, because
numbers in huge fermentation vats, also produce calf it meant that vegetarians could eat cheese made from
chymosin. These genetically modified yeast cells have it without fear of betraying their principles.
been used for the past two decades to produce very Ironically, the Netherlands is one of the few countries
pure rennet enzyme, which is identical to the authentic in which cheese-makers still don’t use it. Although
calf chymosin. Extensive testing has shown that this incredibly late in the day for a country where it was
product of modern biotechnology can be used safely first produced and where cheese production is among
with no risk to health and that it works at least as well the highest in the world, permission for its use was
as the traditional rennin. finally granted in 1992. That said, our domestic cheese
Switzerland was the first country to use this new producers still don’t use it, fearing that the German
chymosin. That was in the late ‘80s, when the public consumers will stop buying our cheese. Germany is
was probably still largely unaware of what sort of one of the Netherlands’ biggest customers, but also the
product it was. Now the people of this country are very country where opposition to anything involving modern
sceptical about modern biotechnology. Meanwhile, biotechnology has been very pronounced since the
this product has been accepted and is used in many beginning of the lobby against gene technology. Despite
countries around the world, while other companies that, the use of recombinant chymosin has also been
have also come onto the market with bovine chymosin, permitted in Germany since 1997. Although cheese
made with recombinant microorganisms. France is one manufacturers in both countries are very reluctant to
of the countries that held back approval for a long time, change, consumers have been buying cheeses made
but then gave permission in 1998 following the “Mad with the recombinant enzyme for years, as many
Cow Disease” episode (the risk of such infection via cheese manufacturers in other countries use it and
rennin cannot be excluded). these cheeses are currently very popular among many
Remarkably enough, the production of chymosin from consumers all over the world. However, the percentage
genetically modified microorganisms has stimulated of end-users who are actually aware that modern
cheese consumption in Israel and the United States. biotechnology has been used to make these cheeses, is
The microbial product has been declared kosher. small. The gentleman’s agreement between the Dutch
Religious Jews can, and therefore do, eat this cheese. dairy companies not to use the recombinant enzyme
Muslims may also eat cheese made with this chymosin, seems now, though, to be on shaky ground, because in
because it is also halal (i.e. meets the Islamic criteria recent times recombinant chymosin has been used on
concerning food preparation). Even Professor Lucas a small scale in Germany. That is probably why its use
Reijnders of the Dutch Institute for Nature and the in the Netherlands will not be long in coming.

74 Part 2: Our daily food and drink

TEXTBOX 4.2. The yeast Kluyveromyces lactis is one of DSM-Gist’s
The PluGbug technology. plugbugs. As we saw above, a recombinant form is used
for making curdling enzyme. This yeast has been used
The recombinant microorganisms which DSM-Gist works for more than 30 years to produce the enzyme lactase,
with primarily, come from normal microbial strains which which is used to convert lactose in milk products into
this company has been using for years on a commercial galactose and glucose. These are sugars that can be
scale to produce enzymes, and thus has plenty of better digested by people who have lactose intolerance
experience and knowledge of it. These microorganisms, (major sections of the population in Asia and Africa
or “bugs” as they are popularly called, have a number are lactose-intolerant), so that these people can also
of advantages which they have been selected on over consume milk products without suffering side effects. At
the years. These include efficient secretion of enzymes the beginning of the 1980s researchers at the company
and the certainty that they are isolated not only the lactase gene, but also the DNA
classified as safe organisms. They which is necessary to express the gene,
BUILD YOUR OWN MICROORGANISM.
are GRAS: “Generally Recognized and therefore the lactase. From this
As Safe”. The recombinant they constructed the so-called gene
microorganisms derived from them expression cassettes, on which they
can be used in the same fermentors can easily record a gene of choice. This
and reprocessing and purification LEGO PLUGBUG cassette can then be accurately inserted
apparatus as the non-recombinant into the genome of the yeast, ensuring
strains, on the understanding that they AS!± with almost complete certainty the
±GR
take place under restrictive conditions, expression of this new gene. They also
i.e. depending on the process, under more performed something similar with the bacterium
or less stringent safety requirements, as prescribed by Bacillus licheniformis and the fungus Aspergillus niger.
the law on the use of recombinant organisms. An added This opened the way to the efficient production of a
advantage of these bugs is that they have been modified whole range of proteins “foreign” to these plugbugs, for
so that they have special food requirements, which example enzymes like chymosin, and pharmaceuticals,
means that in the unlikely event of them getting out of etc. This information about the plugbug concept is taken
the fermentor, they will not be able to continue to grow from the Gist-brocades 1991 brochure ‘Biotechnology,
outside. DSM-Gist has given this technology the trade today and tomorrow’, but more recent publications on
name ‘PluGbug , reflecting the ease with which extra
®
this have appeared since, for example, Groot et al.
genes can be plugged into these bugs . 26
(Groot, Herweijer, Simonetti, Selten, & Misset, 2000).
N.B. In the February 2007 issue of Nature Biotechnology
26
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Chapter 4: Cheese: Biotechnology through the ages 75

 (Cullen, 2007) academic and DSM researchers published promotes muscle regrowth after sporting exertions
the complete sequence of the Aspergillus niger genome. and an enzyme that prevents the formation of the toxic
This genome project also produced new application substance acrylamide in baked and fried products (see
possibilities, namely the production of an enzyme that Textbox 5.3).

4.4. CHEESE RIPENING: divided further into the individual amino acids (about
NOW AND IN THE FUTURE 20 different ones in total form the building blocks of
all natural proteins). These amino acids give the basic
When preparing cheese, as much protein and fat as taste to the cheese, but can also later be converted
possible must be separated out of the milk, and as into volatile (sulphurous) components with a strong
soon as possible this must be set aside to ripen into cheese or cabbage taste.
cheese. Ripening is the complex process required for As already said cheese maturation is a relatively
the development of a cheese’s flavour, texture and slow process, because the enzymes required are
aroma. Proteolysis, lipolysis and glycolysis are the only released when the lactic acid bacteria die and
main biochemical reactions that are responsible for then break open, or lyse. The ripening process is thus
the basic changes during the maturation period. As expensive: the storage of cheese in conditioned areas
ripening is a relatively expensive process for the cheese costs the Netherlands alone more than ten million
industry, reducing maturation time without destroying euros per week! It is hardly surprising then that the race
the quality of the ripened cheese has economic and is on to find new means of speeding up the process
technological benefits. A review of traditional and of cheese ripening. Elevated ripening temperatures,
modern methods used to accelerate Cheddar cheese addition of enzymes, addition of cheese slurry,
ripening is presented by Azarnia et al. (2006). adjunct cultures, genetically engineered starters
As we have already established, the separation is and recombinant enzymes and microencapsulation
activated by adding a milk-curdling enzyme, either of ripening enzymes are traditional and modern
from a genetically modified microorganism or in the approaches to accelerating cheese ripening (Azarnia
form of a naturally-occurring enzyme. Early in the et al., 2006).
cheese-making process starter cultures (Textbox 4.3) An approach used by DSM Food Specialties involves
are added along with rennet. The cultures are a mixture adding extra enzymes. In 1996 the company applied
of lactic acid bacteria, whose composition varies from for a patent on the phenylalanine-aminopeptidase
one cheese to another. Lactic acid bacteria play an enzyme, produced by a non-genetically modified
important role in cheese ripening. Enzymes cleave the mould. This enzyme cleaves the amino acid
proteins into short pieces, peptides, which are then phenylalanine from peptides; phenylalanine is an

76 Part 2: Our daily food and drink

TEXTBOX 4.3. milk enzymes, starter bacteria and their enzymes,
Ripening agents. and enzymes from secondary starter cultures and
moulds. Starter bacteria have an important role in the
Cheese ripening is catalysed by milk enzymes, development of flavour. Because of their main role
coagulant, starter lactic acid bacteria and non-starter in the progressive acidification of cheese, increasing
lactic acid bacteria. All milk components remaining in the number of starter bacteria can result in over-
the curd are involved in the ripening, which involves acidification of the final curd. Attenuated starter
the enzymatic degradation of these components. cultures are used for the purpose of reducing the acid-
In general, the important components in cheese producing ability of the cells without the destruction of
ripening are: chymosin or rennet substitutes, natural their intracellular enzymes.

amino acid that contributes to the taste of the cheese. (Textbox 4.4), or an increase in the salt concentration
The addition of phenylalanine-aminopeptidase to or temperature. Theoretically, the use of these fast-
cheese milk shortens the maturation time of Emmental lysing bacteria can shorten the maturation period of,
and Cheddar. From a marketing perspective, however, for example, Gouda cheese by 75%. The use of this
there is a problem. If the recipe is changed, the technique is, however, currently blocked because of
traditional cheese names can no longer be used. So the previously mentioned protection of type indications
DSM is focusing on the American market of enzyme- such as Gouda, but also because the chance of food
modified cheeses, which are made by grating young containing genetically modified organisms (GMOs)
cheese and heating it up in the presence of taste- being accepted by the consumer is still fairly small
forming enzymes. These enzyme-modified cheeses (Textbox 4.5).
can be perfectly processed into ingredients or A third approach being worked on is the addition to the
flavourings for products like hamburgers and pizzas. starter cultures of lactic acid bacteria that overproduce
On the DSM website 27
one can find their present certain enzymes. One such enzyme is cystathionine-
starter cultures and dairy enzymes. β-lyase. This converts the sulphurous amino acid
In a second approach to accelerating cheese ripening, methionine into methanethiol, a direct precursor of
genetically modified lactic acid bacteria are added to volatile aromas in Gouda cheese. Both approaches
the starter culture. These bacteria can lyse ‘to order’ with genetically modified lactic bacteria have given
and then give up their enzymes to the cheese. This spectacular study results, but to our knowledge neither
order can, for example, be given by the substance are being used yet. The future will decide if and when
nisin (a preservative used in cheese preparation) the general public will fully accept these “classic“
products of modern biotechnology.
27
www.dsm.com/en_US/html/dfsd/home.htm

Chapter 4: Cheese: Biotechnology through the ages 77

 TEXTBOX 4.4. extra quantities of this peptide are made. The nisin
The NICE system. gene can, however, be replaced by an arbitrary gene
X via genetic modification. This is how the patented
Nisin is an antibacterial peptide that is produced naturally NIsin Controlled Expression or NICE system came
by some lactic acid bacteria in order to counteract about. In this system, the expression of gene X and
the development of competing microorganisms. The the production of the accompanying protein X can be
action of nisin relies on the creation of permeable accurately controlled by the addition of more or less
bacterial membranes (Figure 4.1). Since nisin is active nisin (Zhou, Li, Ma, & Pan, 2006).
against perishable and pathogenic microorganisms
such as Clostridia and Listeria, it is used as a natural
preservative, for example in cream cheese and in
NisR P*
signal
Eastern Europe in fruit and vegetable preserves. Once transfer
regulated
a lactic acid bacterium has formed nisin, this peptide Pi gene expression
gene X
nisin
appears to stimulate its own production. Nisin induces nisin NisK
induction
a membrane-bound sensor protein, NisK, to activate
a regulator protein, NisR. This occurs via the transfer
of a phosphorus group (Pi). The activated NisR then
binds to the nisin promotor, P*. A promotor is a piece enzyme X

of the DNA in front of a gene or genes that regulates

the action of this gene or genes. Normally the nisin Figure 4.1. NICE gives control over the production of desired

gene is located behind this nisin promotor, so that proteins such as enzymes.

4.5. THE FINAL QUESTION with more casein protein in their milk. This extra protein
in the milk means that more cheese can be produced
In Chapter 2 we mentioned a development in New more cheaply. As a result of this article, several
Zealand that provoked a strong reaction, namely Members of the Dutch House of Representatives put
the campaign by MAdGE (Mothers Against Genetic questions to the former Minister of Agriculture, such as
Engineering in Food and the Environment). The whether he thought the genetic modification of animals
February 2003 issue of Nature Biotechnology reveals for food production was ethical, whether there should
the scientific background. In this journal Karatzas from be an overall testing framework to weigh up the pros
New Zealand published an article (Karatzas, 2003) and cons of this sort of development, and whether
stating that they produced nine transgenic cloned cows he could prevent these cows or their products from

78 Part 2: Our daily food and drink

TEXTBOX 4.5. labelled as genetically modified (preferred in an earlier
Acceptability of genetically modified cheese. product test) and the other as conventional (neutral
in an earlier product test). A smaller control group
Many European consumers still have rather negative received two cheeses with blind codes. Labelling
attitudes towards the use of gene technology in decreased consumers’ intentions to buy the originally
food production. In 2002 Scandinavian researchers preferred GM-labelled cheese, but still the intentions
published the study “Acceptability of genetically were at the same level as the conventionally labelled
modified (gm) cheese as real product alternative” option. Participants chose two GM cheeses out of five
(Lahteenmaki et al., 2002). The objective of this study possible when given the option to take cheese home
was to examine whether taste and health benefits after tasting. Intentions to buy GM cheese could best
influence the acceptability of genetically modified be explained by respondents’ attitudes towards gene
products when they are presented as real product technology and perceived taste benefits. General
alternatives. Consumers in Denmark, Finland, Norway health interest was also a reinforcer of intentions for
and Sweden (n=738) assessed two cheeses: one was GM cheese with reduced fat content.

being imported into the Netherlands. In Chapter 2 we enough, but the final question remains as to whether or
saw the emergence of precisely that sort of EU testing not Dutch legislation can prevent the resulting products
committee, namely the EFSA. As far as Dutch legislation from being imported, given that the WTO treaties allow
is concerned, genetically modifying and cloning cows to free trade when there are no scientific reasons, for
improve cheese production is definitely not important instance with respect to safety, to ban them.

Chapter 4: Cheese: Biotechnology through the ages 79

 4.6. SOURCES Karatzas, C. (2003). Designer milk from transgenic clones.
Nature Biotechnology, 21(2), 138-139.
Azarnia, S., Robert, N., & Lee, B. (2006). Biotechnological Lahteenmaki, L., Grunert, K., Ueland, O., Astrom, A., Arvola,
methods to accelerate cheddar cheese ripening. A., & Bech-Larsen, T. (2002). Acceptability of genetically
Critical Reviews in Biotechnology, 26(3), 121-143. modified cheese presented as real product alternative.
Cullen, D. (2007). The genome of an industrial workhorse. Food Quality and Preference, 13(7-8), 523-533.
Nature Biotechnology, 25(2), 189-190. Olempska-Beer, Z., Merker, R., Ditto, M., & DiNovi, M.
Groot, G. S. P., Herweijer, M. A., Simonetti, A. L. M., Selten, (2006). Food-processing enzymes from recombinant
G. C. M., & Misset, O. (2000). Enzymes in food and microorganisms--a review. Regulatory Toxicology and
feed: past, present and future. In Stanislaw Bielecki, Pharmacology, 45(2), 144-158.
Johannes Tramper & J. Polak (Eds.), Progress in Zhou, X., Li, W., Ma, G., & Pan, Y. (2006). The nisin-controlled
Biotechnology (Vol. 17, pp. 95-99), Elsevier. gene expression system: construction, application and
improvements. Biotechnology Advances, 24(3), 285-295.

80 Part 2: Our daily food and drink

5 BIOTECHNOLOGY IN THE BAKERY: ON THE RISE!

“You can only really say that something is safe if you yourself are convinced. And we are. The enzymes are not
being tinkered with. And if the enzyme producers are doing that, we’ll know about it.”

This bold statement was issued by Esther Delnoij on 7 May 1994. At that time she was head R&D of a manufacturer
of bread improvers. Like cheese, bread is one of the oldest traditional biotechnological products. In the last decade
of the last century, however, modern biotechnology has also entered the baking industry in the form of recombinant
enzymes as bread improvers and raw materials that may originate from genetically modified crops.

ENZYMES ARE NOT BEING TINKERED WITH

IF THE PRODUCERS WOULD BE DOING IT,

THEY WOULD TELL US!

J. Tramper, Y. Zhu, Modern Biotechnology – Panacea or new Pandora’s box? 81

DOI 10.3920/978-90-8686-725-7_5, © Wageningen Academic Publishers 2011
 5.1. OUR DAILY BREAD are trapped in the dough thus forming the light texture
of well-leavened bread that we know and love.
Our daily bread basically consists of flour, water, yeast
and a little salt. No-one knows exactly when and how BREAD WITH YEAST WAS ‘INVENTED’
yeast was first put into bread to make it rise. It almost BY ACCIDENT IN ANCIENT EGYPT
certainly happened by chance the first time, probably
in the Nile valley at the time of the Pharaohs. What we MMM, LET’S SEE WHAT HAPPENS
do know is that the later Egyptians ate leavened bread WHEN I PUT A SINGLE-CELLED
and that the Old Testament is also clear on this subject. ORGANISM INTO THIS DOUGH...
Here there is a description of bread with or without
sourdough (with or without added yeast) (Textbox 5.1).
As anyone who has ever tried to bake bread knows,
bread leavened with yeast is different mainly in terms
of the texture and structure, but also has a better aroma
and taste. When yeast - a living, single-celled organism
- is added to the dough mixture, the yeast cells grow,
divide and thus increase in number. As the yeast grows,
the cells ferment the sugars in the dough, producing
carbon dioxide among other things. These gas bubbles

TEXTBOX 5.1. hence the name sourdough. After a few days you have
Sourdough. grown a sourdough whose concentration of yeast cells
is sufficient for the purposes of baking bread. To this
Wheat contains by nature different types of so- end, the sourdough must be mixed with more flour and
called wild yeast cells. However, the concentration the mixture then left to stand for a day. The number of
of these yeast cells is so low that it is impossible to yeast cells is however never as high as in the baker’s
get a dough to rise with it. You can however let the yeast that you can buy in the shops. That is a so-called
concentration increase naturally. All you need to do pure culture - one type of yeast grown in large vats
to set this process in motion is add water to flour and (fermentors) in a factory. One gram of this contains
ensure that there is enough oxygen in it. Due to the about ten billion yeast cells, while one kilo of flour only
acetic acid and lactic acid bacteria that also occur contains about 30 thousand yeast cells. Sourdough
naturally in flour, the acidity of the mixture increases, bread is therefore usually less light.

82 Part 2: Our daily food and drink

 5.2. BAKER’S YEAST 5.3. DOUGH

Baker’s yeast can rightly be regarded as one of Wheat dough consists mainly of gluten (a protein
the oldest products of industrial fermentation. The network composed of gliadin and glutenin), lipids
industrial-scale production of baker’s yeast and its (fats), starch and other non-starch carbohydrates.
widespread use probably started with the Viennese This natural raw ingredient can vary tremendously
process developed by Ignaz Mautner around 1846. in quality and also undergoes a great many process
To date approximately 500 different yeast types have steps during bread preparation. Dough is developed
been identified. As a result of its ability to produce as a result of various different processes. First of all,
large quantities of carbon dioxide, Saccharomyces the kneading process breaks up the structure of the
cerevisiae is the most commonly used type in bakeries protein complex, which is formed after flour and water
and for that reason is known as baker’s yeast (Textbox are mixed. The kneading stretches the protein chains
5.2). As far as volume and function are concerned it and lines them up next to each other. During the rising
is one of the most important biotechnological products process of the dough, they form a big protein network,
of all time. Every year more than two million tonnes called gluten. It is important that the gluten proteins are
of baker’s yeast are produced around the world. Most mixed well, as this determines the gas-holding capacity
of the yeast is used to make a large variety of bread of the dough as well as its final volume and firmness.
types. It is also used for pastries, biscuits, crackers For centuries, the variation in the raw ingredients and
and pizzas. the considerable number of processing steps have

TEXTBOX 5.2.
Baker’s yeast.

This photo is an electron microscopic image of

baker’s yeast (Saccharomyces cerevisiae). Yeasts
are unicellular fungi (Ascomycetes), which can survive
in aerobic as well as anaerobic (without oxygen)
conditions. They are important for breweries and
bakeries because of the alcohol and carbon dioxide that
they produce as a result of respiration. Reproduction is
normally asexual by means of budding; the buds are
clearly visible on the cells in the photo.
Source: Getty Images

Chapter 5: Biotechnology in the bakery: on the rise! 83

 made it difficult for bakers to bake consistently good 5.5. ENZYMES
quality bread.
The addition of extra enzymes to the dough has the
CONSISTENT BREAD QUALITY USED
following benefits according to DSM28:
TO BE VERY HARD TO ACHIEVE

IT’Z ALL ZHE ZAME, OKAY!!! • Improved dough handling and process tolerance
• Increased baked volume
• Finer crumb structure
• Improved crispiness and colour
• Softer crumb and extended shelf life
• Replacement of traditional emulsifiers
• Reduced reliance on high-cost ingredients such as
gluten
• Acrylamide reduction (Textbox 5.3)

True enough, enzymes have long been used in malting

and baking, but that was in the form of malt flour
and malt extract. These ingredients are also subject
5.4. BREAD IMPROVERS
to strong variations in quality, so these days bakers
prefer to use well-defined enzyme preparations. The
To make bread quality less dependent on the variations
use of α-amylases (enzymes that hydrolyse starches)
in raw ingredient quality and processing conditions
derived from moulds began in the ‘60s. The α-amylases
during kneading, rising and baking, bakers add so-
(nowadays produced from bacteria) produce dextrins
called bread improvers to their dough. The chemical
(intermediary product in conversion of starch to
bread improver potassium bromate was used until the
sugars) from starch. These are further broken down
beginning of the ‘90s, when it was banned because of
into sugars by the naturally occurring β-amylases in
potential carcinogenic properties. Now ascorbic acid,
dough. This improves the yeast fermentation, thus the
better known as vitamin C, is used with a complex
rising process, and consequently the volume, crust
mix of other substances such as emulsifiers, gluten-
colour and shelf life of the bread. A specific example
reducing agents, sugar, milk solids and a combination
with unexpected benefits is described in Textbox 5.4.
of enzymes. Since the early 1990s enzymes in
Variously sourced proteases (enzymes that break down
particular have been used increasingly in the bakery.
28
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84 Part 2: Our daily food and drink

TEXTBOX 5.3. The enzyme basically converts one of the precursors of
Acrylamide reduction. 29
acrylamide, asparagine, into another naturally occurring
amino acid, aspartate. As a result, asparagine is not
It has been confirmed that a wide range of cooked foods available anymore for the chemical reaction that forms
– prepared industrially, in catering, or at home – contain acrylamide when carbohydrate-containing foods are
acrylamide at levels between a few parts per billion heated. The PreventASe™ enzyme essentially reduces
(ppb, μg/kg) and in excess of 1000 ppb. This includes the formation of acrylamide, by up to 90%. PreventASe™
staple foods like bread, fried potatoes and coffee as well is not required to be listed on the product’s food label,
as specialty products like potato crisps, biscuits, crisp and requires no registration in most European countries
bread, and a range of other heat-processed products. (except for France and Denmark) as it is considered a
Immediately following the initial alarming announcement processing aid. A safety record for review has been
at the start of the century, the food industry within the EU submitted to the French food safety authority AFFSA -
took action to understand how acrylamide is formed in resulting in an approval for the product. PreventASe™
food, and to identify potential routes to reduce consumer has also been approved in Denmark and Switzerland.
exposure. From the onset of the acrylamide issue, the Also in the US, PreventASe™ can be applied without any
efforts of many individual food manufacturers and their further registrations. The FDA reviewed the DSM safety
associations have been exchanged and coordinated under data and provided GRAS notification for PreventASe™.
the umbrella of the European Food and Drink Federation Commission recommendation 2007/331/EC of 3 May
(CIAA), to identify and accelerate the implementation of 2007 on the monitoring of acrylamide levels in food
possible steps to reduce acrylamide levels in foods. These required the Member States to monitor annually in
efforts are also intended to explore how the knowledge 2007, 2008 and 2009 the acrylamide levels in certain
developed by industry might also be applied in home foodstuffs, e.g. bread, potato crisps, instant coffee, etc.
cooking and catering which contribute to more than half At the time of finishing this chapter, July 2010, the results
of the dietary intake of acrylamide. Applying the enzyme of 2008 had just been published in a scientific report30 by
asparaginase in food in order to reduce acrylamide EFSA. The report in general suggests lower acrylamide
has been identified by various institutions as one of the values in 2008 compared to 2007, but soft bread, bread
solutions. PreventASe™ is the first asparaginase enzyme not specified, infant biscuit, and biscuit not specified,
that is used in a commercialised product (DSM). Since showed statistically significantly lower levels. Whether
October 2007 consumers in Germany have been able this represents a trend towards lower acrylamide levels
to buy a Christmas biscuit produced with PreventASe™. over time should become clearer from the reports in the
coming years.
29
www.ciaa.be/documents/brochures/ac_toolbox_20090216.pdf
30
www.nbc.nl/files/EFSA%20rapport%20acrylamide%20monitoring%202008.pdf

Chapter 5: Biotechnology in the bakery: on the rise! 85

 proteins) are normally used to reduce the elasticity of to the Association of Manufacturers and Formulators
the dough in hard wheat varieties. Hemicellulases (or of Enzyme Products (AMFEP), a European industrial
cellulases and pentosanases), whose purpose is to association set up in 1977, the following baking enzymes
break down hemicellulose, are used not only to improve with genetically modified microorganisms were already
the baking properties of robust rye flours, but also to being made and used in 1996: α-amylase, glucose
further optimise the dough properties and the quality oxidase, hemicellulase, lipase, malt amylase, protease,
of wheat bread. A multitude of other enzymes are also pullulanase and xylanase. AMFEP is a non-profit trade
used in bakeries. A recent development, for example, is association which has taken a clear-cut and public stance
the addition of a new lipase (fat-hydrolysing enzyme) as on modern biotechnology since 1995. Textbox 5.5 gives
the first enzymatic alternative for traditional emulsifiers, their policy declaration at the turn of the century. On their
and there are still more in the pipeline. There’s nothing very informative and up-to-date website you can find out
wrong with traditional emulsifiers, except that they all about enzyme regulations in the EC and their latest fact
have an E-number. Food without E-numbers is seen sheet on protein engineered enzymes31. They conclude
to be more ‘natural’ and sells better. Since enzymes this fact sheet with: “Protein engineering is regarded by
are ‘natural’ and have the same effect as an emulsifier, AMFEP as a safe and useful tool in the development of
the latest trend is to add this sort of enzyme. Here too, improved enzyme products and processes that bring
however, modern biotechnology is beginning to play an real benefits to manufacturers, consumers and society.”
increasingly important role, and the next question will AMFEP promotes an open dialogue on the use of this
be: is it still ‘natural’? technology. Most of the bigger enzyme producers are full
(14) or associate (9) members of AMFEP.
5.6. RECOMBINANT ENZYMES On the issue of complete safety in compliance with
internationally accepted standards, various national and
In late 1992 the Dutch Consumer’s Association drew international expert committees have issued guidelines
attention to the fact that much of our daily bread was on how safety assessments should be conducted. These
made using bread improvers that contained an enzyme guidelines are all developed from the basic premise
produced with genetically modified bacteria. This was that the enzymes used in the processing of foodstuffs
confirmed by one of the biggest manufacturers of bread are per se intrinsically safe and that the analysis should
improvers, which means that this enzyme was already focus on impurities and by-products, originating from the
on the market before the Netherlands introduced raw materials or produced during fermentation. These
legislation in July 1993 on so-called “novel foods”. guidelines also apply to the safety assessment of enzymes
By 1997 five recombinant enzymes for use in bread- produced with genetically modified microorganisms.
making were authorised by this legislation. According
31
www.amfep.org/papers.html

86 Part 2: Our daily food and drink

TEXTBOX 5.4. reduction in the cost of transport of bread from bakery
Bread enzymes are also good for the environment. to shop. Approximately 45 percent of the reduction in
energy consumption is a consequence of this reduction
The Danish company Novozymes is one of the world’s in transport. The savings have also led to savings in
biggest enzyme manufacturers. The December agricultural production. Less bread is wasted, so less
2005 issue of their newsletter, BioTimes, contained grain is needed, so less fertiliser is used, meaning less
an interesting article about the added benefits of acidification of the soil. The LCA applies primarily to
Novamyl, one of their registered amylases, often the American situation, but there is a clear message
used in bread making. These added benefits have for the EU, where many member states have a strong
been established by means of a so-called ‘Life Cycle preference for crusty breads like baguettes with a
Assessment’ (LCA). LCA is a methodology that much shorter shelf life.
enables a comparison to be made of the effects on the
environment of alternative production technologies WITH BIOTECH ADDITIVES BREAD
that have the same user benefit. LCA takes a holistic CAN BE PRESERVED MUCH LONGER
look at the business and inspects the whole production MY PRODUCTION
system, from the manufacture of raw materials to the FACILITIES ARE
disposal of waste. ISO guidelines ensure that LCAs IN CHINA NOW...
are performed in a transparent and standard way.
The addition of Novamyl enhances the taste and
texture of breads and also produces a delicious,
fresh bread with a long shelf life (10 to 14 days).
This long shelf life enables bakers to use their
production facilities more efficiently. There is
less need for them to go from one product to the
next and they can therefore prolong production
runs. An LCA demonstrated that besides
considerable reductions in energy consumption
and greenhouse gas emissions, there was also a

AMFEP’s policy statement is designed to prevent ensure that the enzymes used in the processing of
any misunderstanding about what people want foodstuffs are obtained with non-pathogenic and
and what people do. AMFEP members therefore non-toxic microorganisms, i.e. microorganisms that

Chapter 5: Biotechnology in the bakery: on the rise! 87

 have a ‘clean’ safety record, with no reported cases under GMP (Good Manufacturing Practice). All this
of pathogenesis or toxicity that can be ascribed to the and more ensures that enzymes are safe and can be
microorganism in question. The raw ingredients that are safely used. It is therefore hardly surprising that the
used for cultivating the microorganisms are carefully JECFA (Joint Expert Committee on Food Additives
selected so that they do not contain any components of the FAO/WHO) concluded that there is no need
that are harmful to health. Every time a new microbial to limit exposure to enzyme preparations that they
strain is developed with improved enzyme production have evaluated, also with respect to allergy (Bindslev-
capacity or the production conditions are changed, the Jensen, Skov, Roggen, Hvass, & Brinch, 2006); none
potential impact on safe usage is carefully evaluated of them have been allocated an ADI (Acceptable Daily
on a case-by-case basis. Every new strain is checked Intake). You can read more about the authorisation,
for its primary taxonomic properties. If the production labelling and traceability of these enzymes in the
strain contains recombinant DNA, the properties and section on legislation.
the safety record of the donor organisms that delivered
genetic information for the production strain are 5.7. TRANSGENIC CROPS
analysed. The safety of the product is usually backed
by documentation from toxicological safety studies. The advent of genetically modified (transgenic) soya
Consistency and quality are assured by production has revived interest in the use of modern biotechnology
in bakeries. Soya ingredients are used in abundance
GOOD MANUFACTURING PRACTICE in the bakery industry: processed soya beans, soya
flour, soya oil and lecithin. Only the protein-containing
THIS GRAIN PARTICLE fractions of the soya ingredients can carry the
IS OKAY! ONLY 13,786 characteristics of genetic modification, since the DNA
TO GO ... determines which proteins a cell can make. Soya oil
and lecithin are products that don’t contain protein, and
will therefore not usually be altered by the modification.
Bread rises because large protein molecules in wheat
dough form a network, i.e. gluten, that gives the dough
its strength, elasticity and capacity to expand, enabling
the dough to trap the carbon dioxide gas produced by
the yeast. It is primarily the large gluten proteins that
GRAINS

bind together during the mixing and kneading process

to form much bigger polymers. It is for this reason

88 Part 2: Our daily food and drink

TEXTBOX 5.5. • for these reasons we see genetic modification as
AMFEP’s policy declaration on modern biotechnology. an extremely important tool in the production of our
enzymes.
1. AMFEP fully supports and is committed to continuing 3. AMFEP is of the opinion that all enzyme products must
the use of genetic modification (modern biotechnology) be judged on their intrinsic properties and not on the
for the development of improved enzyme-producing basis of the method used to develop the production
microorganisms. This technique offers a whole range organism. Our products – whether or not they are
of benefits and it is important that it is researched produced using genetically modified organisms –
with a view to its use by society as a whole. Genetic are only put on the market once complete safety
modification should be regarded as a logical extension has been established according to internationally
of traditional genetic techniques. accepted norms.
2. The microbial enzymes that are produced by AMFEP 4. AMFEP believes that the ‘right to knowledge’ and
members are used in a wide range of industrial the ‘right to choice’ of the consumers must be
applications. The introduction of genetic modification respected and that an open dialogue is the way to
can offer the following benefits with regard to the win their trust in modern biotechnology. Therefore,
production and/or quality of these enzymes: the AMFEP members are prepared to support their
• greater production efficiency and thus less use of clients and actively inform them if an enzyme is or is
energy and raw materials, and less waste; not produced using genetically modified organisms.
• availability of enzyme products that for economic, 5. AMFEB members will continue to provide clients with
enviro-technical or, as regards production staff, enzymes made using genetically modified organisms
health reasons would otherwise not be available, and to help them tackle consumer concerns. Members
making new applications possible; will not compete on the basis of this technology by
• technical improvements due to higher specificity using claims that support or refute genetic modification.
and purity of enzyme products.

that so much time and effort is spent on selecting and made from non-transgenic seeds, the dough from
breeding wheat varieties with a high gluten content. seeds with one or two extra gluten genes showed a
This is a complex procedure as far as genetics is proportional increase in strength and elasticity.
concerned, because six genes are involved. Despite Researchers in the US are also working on transgenic
that, in 1997, Australian and British researchers wheat with higher gluten levels. In 2004 and 2005
showed that genetic modification has huge potential in trial fields were set up in California and Idaho with a
this area too (Barro et al., 1997). Compared to dough transgenic variety developed by the USDA (United

Chapter 5: Biotechnology in the bakery: on the rise! 89

 States Department of Agriculture). In 2006 baking growers and consumers.
tests were conducted with flour from the first harvest in In the August 2007 issue of Nature Biotechnology
Kansas. Although the results were somewhat mixed, researchers from Pioneer Hi-Bred, a subsidiary of
USDA researcher Ann Blechel concluded nevertheless DuPont, published an article declaring that they had
that they had succeeded in producing a wheat variety identified a key gene in phytic acid biosynthesis. By
that would finally give bakers a competitive edge using special genetic techniques to deactivate this gene
(Anonymous, 2006). However, the question remains (gene silencing) in seed tissue from corn, they were able
as to when bread made from this sort of flour will be to create a maize variety with seeds containing very
sold over the counter to consumers. little phytic acid, but high concentrations of inorganic
The American Bakers Association (ABA), which phosphate. This is good for feed given to cattle with
represents 85% of the major bakeries in the US, one stomach, such as chickens and pigs (see also
would also like to see modern biotechnology being Chapter 7 on meat). The same genetic technique also
used to develop flour with increased levels of vitamins, seems to work in soya beans, suggesting that it might
reduced caloric values, fewer allergens, etc. To help also work in other crops. If that is the case, the ground
broaden acceptance, a new GM wheat needs to has been cleared for producing a grain variety with an
include nutritional improvements for consumers and/or improved dietary value.
improved milling and baking characteristics, according By 2004, the Monsanto Company, a leader in the
to Hayden Wands, director of procurement at Sara production of seeds for genetically engineered crops,
Lee Corp and an official of the ABA. “We are not one had made substantial progress in the development
hundred percent convinced that our customers will go of GM wheat varieties for North America. However,
for a GM wheat unless it has enhanced characteristics,” suddenly in that year, the company scrapped its
Wands told a gathering of representatives from wheat program, in part because of opposition from
agriculture and the technology industry at the North American grain merchants and growers, as well
Biotechnology Industry Organization convention in as concerns that some major foreign importers would
Chicago (Gillam, 2010). According to a researcher reject imports of all American wheat because they
from the University of Melbourne (Bhalla, 2006), wheat could be contaminated with genetically engineered
with improved characteristics (Table 5.1) is expected varieties. In their opinion paper, Miller and Carter
to have a substantial effect on food security and on (2009) plead for a return to this technology. According
our society in general. Bhalla believes that progressive to these authors, greater productivity in wheat farming
and consistent implementation of transgenic crops achieved with improved varieties would confer an
is a basis for an increase in productivity, which has important environmental dividend: wheat is the largest
environmental and economic advantages for both crop in the world in terms of area cultivated (220

90 Part 2: Our daily food and drink

million hectares) and is the second largest irrigated eat our lunch.” The authors can be happy again. In
crop (each bushel produced requires about 40,000 The Wall Street Journal of 7 July 2010 Ian Berry
litres of water on average; it is three times thirstier per reports that the world’s largest seed maker Monsanto
bushel than maize for example); therefore, enhanced and the German chemical giant BASF are starting
productivity would conserve both farmland and to develop genetically modified wheat again as part
water. They conclude their paper with: “Monsanto of an expanded joint venture (Berry, 2010). The
and the United States wheat industry might already declining production in the US has sparked renewed
have been relegated to the position of second mover, farmer interest in developing a stronger variety of
and whoever wins the race to produce desirable wheat.
genetically engineered wheat varieties to the market-
place will enjoy a strong cost advantage and attract 5.8. LEGISLATION
market share in many importing countries. Agriculture
remains an important American industry; one that Chapter 2 describes the role of the European Food
should have learned by now that, if it is slow to bring Safety Authority (EFSA) as the cornerstone of EU risk
the best technology to the table, other countries will analysis concerning food and animal feed safety. The

Table 5.1. Examples of transgenic properties that may help address the needs of a growing world population.

Stress tolerance
Abiotic stress
• Drought tolerance
• Salt tolerance
• Oxidative-stress tolerance
• Improved tolerance for aluminium, boron, cold and heat
Biotic stress
• Resistance to pathogenic fungi, viruses and bacteria
• Resistant to insects and nematodes
Agronomic properties
• Herbicide tolerance
• Improved efficiency in water use
• Hybrids
Quality properties
• Improved grain quality
• Improved nutritional quality – lower phytate levels (Raboy, 2007), higher macronutrient content, greater essential
micronutrient content, and better amino acid composition
• Modified gluten composition for people suffering from gluten allergies
• Special types of wheat with health-promoting nutraceuticals in the grains

Chapter 5: Biotechnology in the bakery: on the rise! 91

 authorisation, labelling and traceability of genetically next to the sales shelf. This must be done in a font size
modified organisms, foodstuffs and animal feed that is big enough to be easily identifiable and legible.
are dealt with by two EU directives, 1829/2003 and
1830/2003, which have been in force since 2004. Not CHOOSING BREAD IS GETTING MORE AND MORE DIFFICULT
all ingredients involving genetic modification need to be
labelled as such; recombinant enzymes, for example, MMM... BREAD WITH REGULAR
which are used as processing aids, do not as yet need CTAGGTACCTGA OR WITH
to appear on food labels. However, this legislation is EXTRA ATCGGCTAGGCAA?
still in the development stage.
EU regulation no. 1829/2003 imposes tighter rules
on the authorisation (safety assessment and permit
allocation) and the labelling of GMOs and genetically
modified food and feed. Genetically modified additives
(substances intended to make a product look better,
last longer, be lighter, etc.) and flavourings also fall
under this regulation. Genetically modified food is food
that consists wholly or partially of genetically modified
organisms or is produced with them or contains
ingredients that are produced using GMOs. This
applies regardless of whether it can be demonstrated In certain cases special features or properties must
that DNA or protein created by genetic modification also be mentioned on the label, especially if, due to the
exists in the end product. The reasons for this are that genetically modified component, the food has a different
the consumer must be in a position to make a well- composition, a different nutritional value or nutritional
informed choice between traditional and genetically effect, a different use or certain consequences for the
modified food. health of certain population groups, compared to the
Since 18 April 2004 this regulation makes it obligatory same food without the genetically modified component.
to label genetically modified food and feed as such, A notification is also obligatory if a food may lead to
when it is delivered to the end user (consumer) or to ethical or religious objections.
an institution. As far as unpackaged products or very A tolerance value (0.9%) has been set for ingredients
small packages are concerned, the information about that contain traces of genetically modified organisms
the presence of genetically modified components must due to unforeseen contamination (during cultivation,
be placed permanently and visibly on or immediately harvesting, transport or storage). In order to be able

92 Part 2: Our daily food and drink

to demonstrate that the presence of the GMOs is it being mentioned on the label, this will be sufficient
unforeseen, companies must be able to produce proof reason to take legal action.
to convince the authorities that they have avoided
the use of GMOs and that it is therefore a case of THE SUPERMARKET
unintentional contamination. GMOs that are (still) not
found to be safe may obviously not be present in the GOLD FIELD MULTIGRAIN
product (zero tolerance). - FRESH FOR LONGER -
PACKAGED ON:
EU regulation 1830/2003 guarantees the availability of 18.04.04
the relevant information concerning genetic modification PRICE €: Ingredients: wheat flour**, water, rye

in all phases of the marketing authorisation of GMOs 1.74 flakes*, barley, baker’s yeast*, yeast
flakes**, sunflower flakes*, vegetable
oil***, soya beans***, wheat flakes*,
and the food and feed produced with them. Information corn grits***, dextrose, emulsifiers
(E472e, E471). Made in a factory where
on the presence of GMOs must be conveyed in writing nuts*** are processed.

at each stage of the chain from “farm to fork” and kept * Product from modern biotechnology
** Product from traditional farming
for a period of five years. Suppliers and purchasers 103 348829993 *** Product from organic farming

(excluding consumers of course) must therefore also be

known. If the supplier doesn’t provide any information,
the regulation stipulates that there is no obligation on 5.9. IN CONCLUSION
the part of the purchaser to mention GMOs on or next
to the product. In the (near) future there are likely to be an increasing
As yet, technical agents such as enzymes do not need number of bakery ingredients originating from
to be labelled as GMOs. Nor do milk, meat or eggs transgenic plants; examples of which are wheat,
from animals that have been fed GM feed need a potato and sugar beet, alongside the currently
GMO label. The same applies to substances that are available soya and corn. We are also seeing a rise
produced by fermentation using genetically modified in the use of enzymes produced with recombinant
organisms (e.g. certain additives or vitamins), but microorganisms as bread improvers. In consultation
where no residues of the microorganism appears in with organisations such as Commodity Boards
the ingredient (contained use). N.B. There is a new and AMFEP, EU governments are working hard on
regulation which is waiting to be approved by the authorisation and labelling rules that are fair and
EC Council, which will introduce changes to these acceptable to all parties. In the end these must enable
exemptions. the consumer to choose between genetically modified
If, during an inspection, it appears that a genetically or non-genetically modified food. This choice will also
modified ingredient has been used in a food without result in more pressure on growers and sellers to fully

Chapter 5: Biotechnology in the bakery: on the rise! 93

 separate (make traceable) product flows. The growing communicating this information to bakers.
application of modern biotechnology in food production We would like to conclude this chapter with a magical
and the relatively rapid changes in legislation in this quote made at the turn of the millennium by the
area, also mean that the bakery industry must keep American culinary writer John Thorne: “Bread is an
pace with and, in particular, capitalise wisely on unparalleled and key source of nutrition among foods.
changing circumstances, especially as concerns public The baked dough feeds the body, but the dough
opinion. It’s good to see national organisations helping itself must be fed by the baker, and the process of
out in this regard by translating the directives into real preparing and baking offers a kind of intellectual and
workplace language and finding user-friendly ways of psychological nourishment.” Keep that in mind!

5.10. SOURCES challenges and opportunities. Trends in Biotechnology,

24(7), 305-311.
Anonymous. (2006, 10 July). Gluten in rich wheat trials. Bindslev-Jensen, C., Skov, P., Roggen, E., Hvass, P., & Brinch,
AgraFood Biotech. D. (2006). Investigation on possible allergenicity of 19
Barro, F., Rooke, L., Békés, F., Gras, P., Tatham, A., Fido, different commercial enzymes used in the food industry.
R., et al. (1997). Transformation of wheat with high Food and Chemical Toxicology, 44(11), 1909-1915.
molecular weight subunit genes results in improved Gillam, C. (2010, May 4). US millers, bakers urge caution in
functional properties. Nature Biotechnology, 15(12), GMO wheat work. Reuters.
1295-1299. Miller, H., & Carter, C. (2010). Genetically engineered wheat,
Berry, I. (2010, July 7). Monsanto, BASF Turn Attention to redux. Trends in Biotechnology, 28, 1-2.
Wheat. The Wall Street Journal. Raboy, V. (2007). The ABCs of low-phytate crops. Nature
Bhalla, P. (2006). Genetic engineering of wheat-current Biotechnology, 25(8), 874-875.

94 Part 2: Our daily food and drink

6“In vino veritas”
WINE: ONE OF THE
OLDEST BIOTECHNOLOGICAL PRODUCTS

Wine is probably the oldest of all biotechnological products, and yet modern biotechnology offers a whole range of
possibilities for its production. Every year approximately 27 billion litres of wine are made from grapes plucked from
about 8 million hectares of vineyard. The “magic” world of wine is currently experiencing a real revolution with its
transformation from a production-oriented to a market-oriented industry. And this revolution depends on innovations
in the area of modern biotechnology! Some of these will be discussed in this chapter.

WINE, ONE OF THE OLDEST BIOTECHNOLOGICAL PRODUCTS

J. Tramper, Y. Zhu, Modern Biotechnology – Panacea or new Pandora’s box? 95

DOI 10.3920/978-90-8686-725-7_6, © Wageningen Academic Publishers 2011
 6.1. WHAT IS WINE? NO WINE, NO LOVE!

According to the shortest possible definition, wine is

YOU DRANK IT ALL?
UHHH
fermented grape juice. According to Wikipedia, wine
is a beverage produced when the juice of grapes
is fermented. Needless to say, the International
Organization of Vine and Wine (OIV) has a little
more to say on the subject32. According to the OIV’s
“International Code of Oenological Practices”,
wine is the beverage resulting exclusively from the
partial or complete alcoholic fermentation of fresh
grapes, whether crushed or not, or of grape must
(juice). The Wine & Spirit Trade Association has also
developed a similar standard definition33: Wine is an
alcoholic beverage, obtained by fermenting freshly Scientific proof of the benefits of wine (for the heart and
picked grapes, the fermentation of which takes place blood vessels) when consumed in moderation, was only
according to the local traditions and practices in the demonstrated at the end of the 20th century. More than
area of origin. a century earlier the first oenologist (Section 6.4), Louis
However, John Baldwinson says in Plonk and Pasteur (1822-1895), came to the same conclusion:
Superplonk (1975) that there is something missing
from the above definitions. Nothing is said about the “Wine can be considered with good reason as the
pleasures of wine: the complex colours, tastes, aromas, most healthful and the most hygienic of all beverages.”
associations. Nothing about the glow a good wine can
give or about the natural (some say lively) character.
Nothing about it being good for you, but above all, 6.2. THE FIRST WINE
there was nothing about the fact that wine can make
you happy. “Wine as a panacea for unhappiness.” The Since time immemorial humans have been getting
pronouncement by Euripides’ Bacchae is completely in microorganisms to work for them. There are indications
agreement on this point: “Where there is no wine, there that wine was already being made from grapes about
is no love, or any other pleasure left for men.” 8000 years ago. Chinese rice wine was made during
the Shang dynasty between 1600 and 1100 BC.
32
www.oiv.int There’s no doubt that there were several “inventors” of
33
www.wsta.co.uk

96 Part 2: Our daily food and drink

wine. However, history names no names, unless Noah 1991. Vinicultural knowledge spread further afield as
can be awarded this accolade. a result of trading with neighbouring countries, arriving
eventually in Egypt. At the same conference it was
Vitis vinifera is the grapevine cultivar most commonly revealed that in 3000 BC wine jugs indeed existed in
used for producing wine grapes and is native to the Egypt. The ancient Egyptians have left behind many
Caucasus. Its geographic central position meant images and other evidence of wine dating back to that
that this vine quickly spread to the rest of the world. time. The Egyptians themselves did not grow grapes,
Until 1991 it was believed that wine making was first so the wine must have been imported. At that time not
performed approximately 3000 BC. In 1991, however, only was the art of winemaking already established,
Virginia Badler of the University of Toronto presented but there were thus also a lively trade in wine. In
her research results at a conference of wine experts in Mesopotamia, in the present area of land in Iraq that
California. She had examined a Persian earthenware lies between the Euphrates and the Tigris, 7000-year
amphora from 3500 BC which had a red stain on the old pitchers containing traces of wine have meanwhile
bottom. Infrared spectroscopy revealed that the stain been found in archaeological digs.
contained tannin and tartaric acid among other things. Grape vines reached Greece about 2000 BC.
Both substances occur in wine. Previously, scholars Amphoras (Textbox 6.1) and a winepress found on
from Israel had discovered that grapes had indeed Crete date back to 1500 BC. The art of winemaking
been grown there in 3500 BC. The difference between then spread from Greece to Italy, France and Spain.
wild and cultivated grapes is easy to identify by the These three countries were once the biggest producers
different forms of grape seeds. So clearly wine was in the world, until the United States, Argentina, Chile,
already made 500 years earlier than thought until South Africa and Australia joined them.

TEXTBOX 6.1. of volume for liquids (approximately 23 litres). Since an

The amphora and traditional Greek wine: retsina. amphora was too porous a vessel in which to keep wine,
resin was added to the wine to “seal” the amphoras.
An amphora is a jug, with two handles, Because resin also vastly prolonged the shelf life of
that narrows to a point at the bottom. wine, the Greek viniculturists continued to add resin to
Amphoras were generally used at that their wines. The Greeks got used to the resin taste and
time to store and transport liquids like even became attached to it, whereby retsina became
oil or wine, but also to store grain. Apart the most well-known and best-loved Greek wine and is
from its use as a name for a jug, a still made in abundance. As an outsider you either hate
Greek amphora was also a measure it or you love it.

Chapter 6: Wine: one of the oldest biotechnological products 97

 The entire wine industry is presently noticing the best they could come up with was a semi-magical
(negative) effects of global warming. It seems explanation. With the scientific knowledge we
probable that this global warming will shift the wine possess today about how complex these processes
boundary further and further north. Winegrowers in are, we can only sit back and admire these so-called
the three biggest European wine countries, France, “primitive” people. Time and time again they managed
Italy and Spain, are already moving to cooler areas to develop something with which they were not only
and in the Netherlands a growth in the number of able to make wine in a reasonably reliable way, but
commercial vineyards is clearly visible. Maybe modern also other biotechnological products like cheese,
biotechnology has the solution to the problems created bread, beer and soft leather. All without any scientific
by this climate change (Section 6.9).
34
understanding of the processes.

6.3. ALCOHOL AS A STIMULANT 6.4. THE SCIENTIFIC DISCOVERER:

LOUIS PASTEUR
In the previous chapter on bread we described
how yeast produces carbon dioxide when the yeast Scientific understanding of fermentation processes
cells grow on the sugars in the dough. There is, really only started in the second half of the nineteenth
however, another by-product, albeit in relatively small century when in 1867 Louis Pasteur discovered
quantities, because the bread yeast strains used are a number of undesirable microorganisms which
mainly selected for their capacity to produce carbon spoiled the fermentation of wine and beer. In fact,
dioxide. For most other applications, that by-product microorganisms had been discovered a few hundred
- alcohol - is much more interesting and is even years earlier in 1676 by Antonie van Leeuwenhoek.
the main component. This ‘stimulant’ was probably This Dutch scientist had designed a primitive
discovered back in prehistoric times, as mentioned microscope and studied what he himself called ‘the
above, when people began making a sort of wine smallest animals I have seen thus far’. Pasteur was
and beer. It was initially produced domestically, but asked by Napoleon III to find out why wine, already an
later developed on a semi-industrial scale in sheds important export item for France at that time, spoiled
behind inns and public houses, and in monasteries. during transportation to consumers abroad. His studies
Much later still it was produced on an industrial scale resulted in three simple, but oh so very important,
in wineries and breweries. What’s certain is that the guidelines for making wine, and for fermentation in
first winemakers and beer brewers had no idea how general.
or why these fermentation processes took place. The The first guideline, which now seems so obvious
to us, is hygiene. Wine can only be prevented from
34
www.gmo-compass.org/eng/database/plants/73.grape_vine.html

98 Part 2: Our daily food and drink

rapidly turning into something else, usually vinegar, if sugars to carbon dioxide and water, alcohol is
it is produced and stored in clean containers, as well primarily created. So, an abundance of oxygen
as being sealed off from air and thus from the many stimulates the growth of the cells and reduces alcohol
microorganisms floating around in it. Interestingly formation.
though, making vinegar from wine is also an old
biotechnological process using acetic acid bacteria. 6.5. HOW IS WINE MADE?
The second guideline is the use of one of Pasteur’s
innovations, namely the process that bears his name, Wine is the product of fermented grape must or juice.
pasteurisation. This process is based on the necessary The diagram below shows a very simple overview of
hygiene procedure for extending shelf life. When wine, the red and white wine production routes. However,
beer or other products like milk, are heated and stored it is important to realise that many hybrid forms and
for a while at 75º C, most of the pathogenic and food- specific details have been introduced by knowledge,
spoiling microorganisms are killed, so reducing the art and tradition. Harvested grapes are processed
possibility of a loss of quality, and spoilage, as a result immediately. Crushed whole grapes, called must, are
of undesirable fermentation. used at the beginning of the fermentation process for
making red wine. Colourings, aromatic substances and
WINE INNOVATION: PASTEURISATION flavourings in the skins are extracted in this process.

HEY DAD, LOOK! I’M EXTENDING White wine is produced by first removing the skins in a

THE SHELF LIFE OF YOUR WINE! press before the fermentation. Because the colouring
substances are present in the skin, white wine can also
be obtained from red grapes. Thus only juice is further
processed, starting sometimes with clarification. The
fermentation can be set in motion by inoculation with
a commercial strain of the yeast Saccharomyces
cerevisiae. It is also possible to conduct a natural
or non-inoculated fermentation by using the wine
yeasts that are present naturally on grapes or that
are airborne via the natural flora. At the start of such
Pasteur’s third scientific contribution consisted of a natural fermentation, there are also many wild, non-
understanding the importance of oxygen in the Saccharomyces yeasts and bacteria present, but by
fermentation process. By growing yeast without the end of the process Saccharomyces cerevisiae has
sufficient oxygen for the complete conversion of outnumbered them.

Chapter 6: Wine: one of the oldest biotechnological products 99

 in order to remove undesirable substances that cause
cloudiness, bitterness or acidity.
Depending on the winery and the quantity of
red grapes press fermentation clarification bottle fermentable substances still present, the wine
sometimes undergoes microfiltration so that it enters
the bottle sterile. As Pasteur established, this is
important in terms of shelf life. However, it should be
white or red press clarification fermentation bottle noted that generally the wine does not undergo a heat
grapes
treatment. This would destroy too many aromas and
Aside from climate, grape and yeast variety, and soil flavours and would seriously damage the character of
type, the metabolic activity (metabolism) of other the wine. Good hygiene is the watchword.
microorganisms contributes significantly to the taste
and aroma of the final wine. Lactic acid bacteria, for 6.6. ENZYMES ARE THE SOLUTION!
example, play an important role in the formation of
flavours and aromas. These bacteria convert the Over the last few decades enzyme manufacturers
sharp-tasting malic acid (malate) in the must into have introduced a series of enzymes onto the market
much milder lactic acid (lactate) and carbon dioxide. for all manner of applications in wine production.
This process is called malolactic fermentation and is There are, for example, enzymes called pectinases
responsible for the “fatty” character of the better wines. and hemicellulases that stimulate the extraction of
This conversion is also especially important in wines juice during the crushing and pressing. The cell walls
that would otherwise be too acidic. of grapes consist largely of pectin and hemicellulose
After fermentation, wine undergoes a number of further which are broken down by these enzymes. They
minor or major processes. Red wine first goes through dissolve, as it were, so that the fruit juice in the cells is
the press to remove the skins and other solids, and more easily released.
is clarified if necessary. Many wines are then left to Enzymes are also added for a more efficient release of
mature before being bottled. That can be for a short flavours and aromas, thus allowing the characteristic
time in large tanks, or for years in oak barrels, and aromas of grapes to develop to their maximum potential.
many other configurations in between. During this time This is done primarily with the aim of increasing the
all sorts of chemical changes occur that increase the concentrations of free terpenes in wine. So, although
complexity of the wine, i.e. the bouquet, the flavour you can’t make a bad wine good, you can make a good
becomes more diverse. The wine can be processed wine better. These enzymes provide the winegrower -
further with adsorbing substances, such as proteins, the most traditional of all craftsmen - with all kinds of

100 Part 2: Our daily food and drink

new tools for enhancing the quality, consistency and 6.7. CHAMPAGNE WITH A FLICK OF THE WRIST
stability of his product.
A classic problem in winemaking is the clarification, i.e. Champagne is the most prestigious of all the sparkling
making the wine clear. After every harvest winemakers wines. It is made according to the quality control rules
are confronted with the issue of whether the wine of its appellation: in a very limited region, from specific
will become clear and therefore be easy to filter. The grape varieties and using very well-defined procedures.
cloudiness and accompanying filtering problems are The production process for sparkling wines usually
chiefly caused by the presence of pectins and glucans. consists of two main steps. First, a basic wine is made.
These are large molecules that are present in the wine This has certain specific properties, for example, a
forming solid aggregates which result in a cloudy wine. moderate alcohol content by using early plucked grapes
The gummy glucans in particular can make filtering which still contain very little sugar. After the mixing
problematic. Some of these glucans are associated of various basic wines, if this is required, a second
with the Botrytis cinerea mould, which makes the fermentation is initiated. For champagne this is done
grapes decay. Even if only a very small percentage in the bottle that the consumer purchases. This second
of the grapes used is partially rotten, this can make fermentation is brought about by the addition of the
the resulting wine incredibly difficult to filter. The wine liqueur de tirage, a sugar and yeast mixture. The bottle
can remain cloudy for weeks, even months, which is is then fitted with a crown cap and stored horizontally at
detrimental to the quality. Enzymes can offer a solution 11-12 °C. The second fermentation process only takes a
here too. By adding a mixture of pectinases and few months, but the champagne is then left to mature for
glucanases immediately after alcoholic fermentation, two to eight years, or even longer for top champagnes.
the pectins and glucans are broken down and the wine
quickly clarifies, thus eliminating filtration problems.
Many of the enzymes used are products of
modern biotechnology, because they are made
with recombinant microorganisms. The enzymes
themselves are authentic, i.e. no different from those
produced by the original organisms. Furthermore,
since they are used in relatively small quantities, as The next stage is the remuage: the turning of the bottles
auxiliaries, there is no obligation to mention them on in special racks to collect the yeast deposits in the bottle
labelling (this may change in the future; see previous neck at the crown cap. The bottles are slightly turned
chapter on bread). Some of these enzymes, and others with a rapid motion several times a month and returned
too, are also used on a large scale to make fruit juices. to an increasingly vertical position, with the neck pointing

Chapter 6: Wine: one of the oldest biotechnological products 101

 downwards. The length and intricacy of this procedure is almost everywhere. For the disgorging, the bottles, with
determined by various factors, in particular the settling the crown cap still pointing downwards, are inserted a
properties of the yeast. A month at least is usually little way into a low-temperature bath so that only the few
required. The duration, the storage, and the labour- millimetres of champagne containing the yeast in the neck
intensive nature of the remuage process determine the are frozen. The bottle is placed upright and the crown
cost. So a great deal of effort has been put into simplifying cap with the frozen champagne/yeast prop is removed.
this step, including automation. The space left is topped up with the liqueur d’expédition,
Ten years before the end of the last century, an ingenious which is not just a straightforward sweetener, but a
method was devised by the famous champagne house method of further improving the champagne. The quality
Moët et Chandon. Live yeast was ‘immobilised’ in little 1 of this liquid, the subsequent maturation, the character
mm balls. The immobilisation process that they designed of the wine, the quality of the sugar, and the recipe, are
for this purpose is such that the immobilised yeast behaves all important for the quality of the end product. Finally,
in the same way as the normal, free yeast. However, the bottle is hermetically sealed with the special cork, the
now after fermentation in the bottle, the remueur simply wire collar and the cap. The final result: a party!
has to turn the bottle upside down and the yeast balls
roll along to the crown cap. In short, with the flick of a 6.8. MANIPULATION OF WINE YEAST
wrist the process is complete, resulting in enormous cost
savings. And yet, as far as we know, this process is still Winemaking may well be older than documented history,
not used for making champagne. The employee unions, but there is still room for improvement in many areas
who obviously fight for employment opportunities, have of the process. This is where the application of modern
worked hard to ensure that champagne made in this way biotechnology has so much potential, and why modern
is not allowed to carry the champagne label. The process winemakers are so keen to find out exactly what it has
is sold to producers of sparkling wine from other regions to offer. There are research groups all over the world
(Spain) which don’t have this problem. working on the genetic modification of yeasts in order to:
N.B. The immobilisation of enzymes is a technique
that was developed in the 1960s and 1970s, in order • improve the settling of wine yeasts;
to facilitate the more efficient and more effective use of • optimise the balance between acid and alcohol levels;
these biocatalysts. Immobilisation of entire cells followed • intensify the colour;
in the 1980s. Now many of these immobilised biocatalysts • make the wine “fuller” or “fattier”;
are used on a large scale in industry. • prevent undesirable substances entering the wine;
Nowadays the rest of the process, i.e. the disgorging, the • increase the concentrations of health-improving
removal of yeast, refilling, and recorking, is automated components.

102 Part 2: Our daily food and drink

Yeasts that easily flocculate and/or easily precipitate wine produced with this genetically modified yeast
are worth their weight in gold for sparkling wines, as has more of the desirable volatile acids and better
outlined above for champagne. Every yeast type has colour properties than wines produced with traditional
a set of genes that causes flocculation. But this set is yeasts plus lactic acid bacteria. An analysis of volatile
often not switched on. At the INRA35 (French National components and a sensory evaluation has shown that
Institute of Agronomic Research) in Montpellier, industrial production of wine with the recombinant yeast
researchers have made a recombinant champagne strain is suitable for the commercial production of quality
yeast with its own specific switch to activate the wines.
flocculation genes in these traditional yeast cells. In
BIOTECHNOLOGY CAN TURN
principle, therefore, it will be possible in the future for a
POOR WINE INTO GOOD WINE
winemaker to flocculate the yeast to order.
At the same institute, researchers also inserted genes BUT CHANGING THE LABELS
from lactic acid bacteria into wine yeasts, enabling these
DOES THE TRICK AS WELL!
to convert malic acid into lactic acid and carbon dioxide,
i.e. perform the so-called malolactic fermentation without
using lactic acid bacteria. Malolactic fermentation is often
problematic, because lactic acid bacteria do not thrive €2,-

well in alcohol. Alcoholic and malolactic fermentation VINO

EL
CHEAPO EXC
can now take place simultaneously and can be executed LUS
IVE

by one and the same recombinant yeast. The use of

€25,-
this type of yeast strain has now been approved by the
American FDA (Food and Drug Administration), and Although the name of the quoted writer in Textbox 6.2
has been granted GRAS status (Generally Recognized is unknown to us, we largely agree with her/him. We
As Safe). According to the literature, it is already being also care a great deal about the future of wine, but in
used commercially in Moldavia and the US. The yeast our opinion it will never be just another manufactured
developed by Springer Oenologie (Textbox 6.2), part beverage. No two wines are the same, not now,
of the Lesaffre Yeast Corporation, can induce the not ever! Our line in the sand bans poor quality and
alcoholic as well as the malolactic fermentation in a unsafeness. That will guarantee a blooming enterprise,
matter of five days; so saving the wine producers time with no risk of destroying the whole venture.
(AgraFood Biotech, 25 June 2007, p. 7). According to There have been other fascinating attempts by
recent research conducted by Canadian scientists, biotechnologists to make good wine from poor wine
using recombinant yeasts. Good wine tastes full-
35
www.inra.fr

Chapter 6: Wine: one of the oldest biotechnological products 103

 TEXTBOX 6.2. a stage at which wine can be at risk. Also, the resulting
GM yeasts: the next battleground? wine is less likely to contain biogenic amines which are
produced by the bacterial malolactic fermentation and
On the wineanorak website 36
it is predicted that which can have negative health effects. In the USA
the next battleground in the wine world will be the yeasts are classified as processing agents, and thus
controversial use of genetically modified (GM) yeasts wines made with this yeast would need no declaration
in winemaking. We quote from it: that they contained GM ingredients. This allows GM
“Plenty of these genetically modified strains already yeast to enter winemaking ‘under the radar’, with
exist in laboratories around the globe, but they haven’t consumers or advocacy groups none the wiser. In
previously been commercialized because of the many other countries, such as New Zealand and
negative reactions of consumers to GM food products. Australia, the regulations are more stringent, and yeast
The scientists are busy engineering beneficial traits is considered as part of the ingredients of wine.
into wine yeasts even though they know they won’t be So is anyone making wine using this GM yeast? If they are,
useful for commercial winemaking for the foreseeable they aren’t telling anyone, for understandable reasons.
future … Now, however, a GM yeast strain, called In response to the commercial approval of ML01 in the
ML01, has been commercialized and is authorized USA, the Australian Wine Research Institute has issued
for use in the USA. This yeast, made by Springer a statement declaring that no GM yeasts will be used in
Oenologie, has been the recipient of two extra genes Australian wine for the foreseeable future. But because
(known as transgenes). The first is a malate transporter it is so much easier to produce yeasts with desirable
gene from another yeast, Schizosaccharomyces properties by GM technology (and there are some
pombe, and the second is the malolactic enzyme gene traits that are impossible to select for by conventional
from Oenococcus oeni, the main bacteria responsible breeding), research continues apace globally on
for the natural malolactic fermentation that occurs in GM yeast technology. So what’s the big deal? Aren’t
many wines after alcoholic fermentation. This yeast GM microbes used all the time? … Supporters of the
is therefore able to carry out malolactic fermentation technology argue that what they are doing by developing
(normally done by bacteria) at the same time as GM yeast strains is not with the intention of creating fake
alcoholic fermentation. There are several advantages wines, but with a view to unlocking the latent flavour
to this. The first is that processing wine becomes much and aroma potential of grape must by using yeasts with
faster. The second is that there is less risk of wine special properties. One yeast researcher has even gone
spoilage because there is no delay between alcoholic on record as stating that the best wines are still to be
fermentation and the onset of malolactic fermentation, made, and that this technology is one way forward. What
do I think? As a scientist who cares a great deal about
36
www.wineanorak.com/GM_yeasts.htm

104 Part 2: Our daily food and drink

the future of wine, I favour a cautious approach: if GM knock the elegant science involved in engineering new
yeasts become widespread, the danger is that wine will wine yeasts, I’m afraid I’m going to voice my disapproval
be seen as just another manufactured beverage. If we at the use of GM yeasts in wine. I think it’s time we drew
kill the ‘naturalness’ of wine, we run the risk of destroying a line in the sand and banned the use of GM organisms
the whole venture. So although it rankles with me a bit to in winemaking.”

bodied, a property that stems to a large extent from including protection against heart attacks. However,
the glycerol in wine. However, the strict AOC rules do their real effectiveness in this area is still questionable,
not allow glycerol to be added. But who can protest according to Katan37, an expert in the field of human
if a “new” yeast produces a little more glycerol in the nutrition and the person who has for years explained
wine? This yeast now exists, although a lot more to the general public what is fact and what is fiction
manipulation is required, because it also produces in the claims made by the food industry. In his view,
quite a few undesirable by-products, such as acetic red wine is probably no better than vodka at protecting
acid. us against the risk of heart attack. Nonetheless, we
Saccharomyces cerevisiae, the wine yeast par see wine yeast being genetically modified so that the
excellence, can convert arginine, one of the most fermentation produces more resveratrol (Pretorius &
prevalent amino acids in must, into ornithine and uric Bauer, 2002).
acid during fermentation. However, S. cerevisiae does There is an enormous diversity of strains of the yeast
not immediately use all the uric acid produced. “Free” Saccharomyces cerevisiae (Pennisi, 2005) and the
uric acid reacts spontaneously with the ethanol in wine genetic properties of wine yeasts differ greatly from
to create a new substance called ethyl carbamate. place to place. As a result the variation in commercial
Unfortunately, ethyl carbamate is carcinogenic. yeasts is also very great - accordingly there are as
Research has shown, however, that an industrial wine many wines as yeasts! There is still, however, a lot
yeast can be genetically modified so that the production of research and optimisation to be done. The above-
of ethyl carbamate in Chardonnay is reduced by almost mentioned article by Pretorius and Bauer gives a
90 per cent. Complete removal also seems feasible. In good overview of the objectives of current research
fact, Japanese scientists have already achieved this in this area. Basically it all comes down to improving
in sake. the efficiency of the fermentation process and the
Red wine contains polyphenols, including quercetin subsequent steps, suppressing microbial decay, and
and resveratrol that come from the grape skins. These above all increasing the health benefits and, of course,
substances are attributed with favourable health effects, improving the flavour, colour and the bouquet of wine.

37
www.falw.vu.nl/en/research/health-sciences/people/martijn-katan/index.asp

Chapter 6: Wine: one of the oldest biotechnological products 105

 6.9. MANIPULATION OF THE GRAPES reported at the beginning of 2009. Researchers
at the University of Illinois at Urbana-Champaign
Vines are classified under the Vitis genus. This created a genetically modified grape, called Improved
genus has two subdivisions, namely Euvitis and Chancellor, with resistance to the herbicide 2,4-D
Muscadinia. One single type, Vitis vinifera, has its (Textbox 6.3). Chile and South Africa are also very
origins in Europe, in the Caucasus, while in China active in this field: South Africa for instance in the
there are more than 30 indigenous vines, and in North context of the “Grapevine Biotechnology Program” by
and Central America more than 30 types have been the Institute of Wine Biotechnology in Stellenbosch
characterised. V. vinifera is the most cultivated species and Stellenbosch University (Stellenbosch University
and approximately 5,000 cultivars of this species News, 31 August 2006). Vines as well as yeasts are
are grown on a commercial scale. The same goes being genetically modified in this program and field
for grape cultivars as for yeasts: there are as many trials started in 2006.
wines as there are cultivars. In short, no two wines The subject of GM grapes is discussed extensively
are the same! Initially new cultivars were obtained in a second review article by Pretorius (Vivier &
primarily by randomly selecting natural mutants with Pretorius, 2002). The authors present an overview of
increased yield and/or better grapes for winemaking. those properties that are desirable in vines and that
In the second half of the 20th century this was done can be worked on at the present time with the help of
primarily via a more targeted selection of clones, but recombinant DNA technology. First in line is increased
in 1994 the first field trials took place with genetically resistance to infectious diseases caused by moulds,
modified vines . In the EU so far seven field trials
40
bacteria and viruses. This is to be expected given
with GM grape vines have been executed. A notable that grapes are among the most heavily sprayed of
example with gene-modified fungal-resistant grape all crops, requiring an average of 12 applications in
vines started in 1999 in two areas in Germany, the a season (DeFrancesco & Watanabe, 2008). Second
Palatinate (Pfalz) and Franconia (Franken). The field is higher stress tolerance, i.e. to drought, oxidation
trials were planned to last for 10 years, and examined damage, temperature and osmotic and other abiotic
mainly the varieties Riesling and Chardonnay, stresses. Finally, improved properties concerning
for which it had not been possible previously to quality, such as sugar content and colour.
breed fungus-resistant vines. As result of fear and In their paper DeFrancesco and Watanabe address
political pressure the trials were suspended at the the following question: “With the genome of the
beginning of 2005. In the USA the number of trials grapevine in hand, how likely are oenologists and
is now around sixty. A region-specific example was winegrowers to resort to genetic engineering to tackle
the problems facing viticulture?” They start by saying
40
www.gmo-compass.org/eng/database/plants/73.grape_vine.html

106 Part 2: Our daily food and drink

that the complete genome sequences of two grapevine genome of the two cultivars are over 150 genes for
cultivars now grace the public genome databases, aroma and flavour, three times the number found in
bringing the latest sequencing technologies (see other flowering plants. This knowledge should be very
also Chapter 13) to bear on one of the oldest uses encouraging to grape geneticists and breeders who are
of biotechnology – winemaking. “The grapevine considering the use of recombinant-DNA technology.
genome now joins the august group of completed However, the fear of a public outcry against GM grapes
plant genomes, being the first freshly fruit crop … may continue to stop transgenesis of grapevines from
to be sequenced.” Found amid the sequence of the taking hold. The authors end by quoting Marc Fuchs,

TEXTBOX 6.3. techniques to do so. … Of the eight Chancellor grape

GM Grapes Raise Hopes for Midwest Wine plants eventually developed through this process,
Industry. 38
three retained the herbicide resistance gene. Cuttings
from the Improved Chancellor plants, along with a non-
On 1 January 2009 H. Sterling Burnett reported in modified Chancellor used as a control, were sprayed
the Environment & Climate News on the genetic with relatively high amounts of 2,4-D. The modified
modification of Midwestern grapes. He wrote: “One of Chancellor grapes proved resistant to the herbicide.
the most effective, widely used herbicides in the United … Once the grapes have been found safe to eat, the
States – known as 2,4-D – has a serious drawback: It research team will have to work with a grape grower
devastates grapes. That makes it very difficult to raise to produce a wine using Improved Chancellor. Even
grapes in the Midwest, because 2,4-D is widely used then, environmental activists are likely to mount legal
on popular staple food crops including corn and wheat, challenges to the distribution of the grapes, further
and it can harm grapes up to two miles away from its delaying their introduction into the marketplace,
point of application. … In the aftermath of an accident experts say.”
spill of the pesticide, the United States Department of Bearing in mind the “loud outcry” concerning GM
Agriculture (USDA) found a soil bacterium with a gene Grapevines of the Institute of Science in Society39 we
that allows it to break down 2,4-D. Building on these reckon that the latter statement will prove to be true.
findings, in 2002 Robert Skirvin, a plant biologist at The outcry reads: “Clearly these new developments
the University of Illinois, secured permission to use the are crying out for GM labelling at the very least, and a
gene and transfer it into the Chancellor grape. Skirvin clean sweep of the regulatory regimes would not come
and his colleagues used standard genetic engineering amiss.”

38
www.heartland.org/publications/environment%20climate/article/24364/GM_Grapes_Raise_Hopes_for_Midwest_Wine_Industry.html
39
www.i-sis.org.uk/GMGrapevines_and_ToxicWines.php

Chapter 6: Wine: one of the oldest biotechnological products 107

 molecular biologist at Cornell University since 2004 in an article published in 2005 (Pretorius & Hoj, 2005).
after working in viticulture for twenty years in France. According to them, the technological possibilities of
He acknowledges grower resistance to GM wines and modern biotechnology will cause a paradigm shift: the
notes: “The wine industry is not very receptive to any current process-oriented wine industry will become
innovation.” However, over the past five years, he has consumer-oriented, and thus market-oriented. This
detected a shift in mindset among the people with market demands wine that stimulates all our senses
whom he interacts. “Most growers are convinced that (except hearing, although … think of the crystal clear
science should move forward in case scientists have sound of clinking wine glasses), that is beneficial for
an interesting plant to offer the industry. They could our health and that can be produced sustainably.
then make the choice to use it or not,” he says, but Moreover, it wants wines that continue to be shrouded
Fuchs is optimistic that the path to commercialisation in the undefined mystique we associate with them
is opening up. And so are we. In the above-mentioned (Bisson, Waterhouse, Ebeler, Walker, & Lapsley,
article, Vivier and Pretorius also look at the obstacles 2002). For that, no two wines should ever be the same,
that must be overcome if genetically improved vine that is our message.
cultivars are to be marketed. For example, in the area
of: 6.10. WINEMAKERS RAISE THEIR GLASSES TO
BIOTECHNOLOGY
• science and technology;
• legislation; This was the headline to an article about wine in the
• intellectual property and patents; July 2006 newsletter of LIS Consult, a small Dutch
• politics and economics; consultancy office in the field of biotechnology.
• marketing; Readers can deduce from this headline that
• tradition and culture; winemakers have something to celebrate as far as
• social acceptance. biotechnology is concerned. The same conclusion
could be derived from the headline to an article in
What is clear from these very informative review AgraFood Biotech of 13 February 2006: Wine industry
articles by Pretorius et al., and also from other earlier gradually accepting GM. Not surprisingly, since both
articles from this group, is that the authors are in favour articles use the same sources41, 42. The LIS Consult
of using modern biotechnology in the wine industry. article reads as follows:
However, the articles also show that in 2002 there “Winemaking is steeped in tradition. Connoisseurs
were still many obstacles on the road to commercial place a great deal of value on the production conditions
implementation. Their tone was clearly more optimistic 41
www.checkbiotech.org
42
www.whybiotech.com

108 Part 2: Our daily food and drink

(soil, climate, aspect of the slopes, etc.), and the also been reported in Chile, Eastern Europe and South
maturation and the development of taste and bouquet. Africa. Research is even underway in Italy. And in the
From a technological point of view, much has changed Colmar region of France, small-scale field trials have
in recent decades in the process of winemaking, but already been in progress for some years. In addition to
the cultivation of the grapes and the variety of grape the development of grape cultivars that are resistant
being grown has continued virtually unaltered. Now, to a number of very common diseases, research is
though, changes are afoot in that area. Until recently also being conducted into whether it is possible to
winegrowers, both in Europe and the US, were fierce grow ‘healthier’ grapes. In recent years much has
opponents of the advent of biotechnology in grape been written about health-promoting substances
cultivation. But in a lengthy article on the website of particularly in red wine, e.g. sterols, polyphenols and
the Canadian Council for Biotechnology Information antioxidants, which are micronutrients that may play a
there is mention of a clear policy change among a role in preventing cardiac disorders. The aim is to try
number of leading organisations in the wine sector. to increase concentrations of these components using
As an example, Winetech, a professional association biotechnology, and to make wine even ‘healthier’.
of South African winemakers, recently accepted a
resolution expressing support for the introduction BIOTECHNOLOGY MAKES WINE HEALTHIER
of biotechnology ‘to help in the development of the
tradition and science of winemaking’. The policy is I GET MY WINE
aimed at ‘promoting innovative research and dynamic FOR FREE FROM
science in a responsible and intelligent way’. In HEALTH INSURANCE?
California the representatives of the wine counties
have expressed their support for biotechnology
because they expect it to play a key role in making
grape cultivation sustainable in the long term. This,
in spite of the fact that some counties have accepted
non-GMO resolutions. Another sign that changes
are afoot comes from wine countries around the
world. Australia, Canada and various West European
countries have started field trials. And according to C.
Ford Runge, a professor and director at the University On top of all that, work is also being done on improving
of Minnesota Center for International Food and the yeast types that are used to make wine. For
Agricultural Policy, lab and greenhouse trials have industry, the reduction in maturation time, which

Chapter 6: Wine: one of the oldest biotechnological products 109

 translates into cost savings, is what appeals most. As We conclude this chapter with a free interpretation of a
with many other reports on its website, the Council for piece from the article by Pretorius & Hoj (2005).
Biotechnology Information also tries to present this The image of wine as a harmonious blend of nature,
area of application in a good light. Whether or not the art and science has long been a source of tension
consumer will accept all of this is obviously the million between tradition and innovation. At the beginning of
dollar question, because wine is for many consumers a the 21st century this tension has increased as a result
hand-crafted product, where tradition plays a key role.” of the innumerable promising possibilities that modern
It is clear that authorisation committees such as the biotechnology now offers for the genetic modification
EFSA will continue to pour over this sensitive issue in of grapes and yeast. The greatest challenge is to make
great detail. All that remains is the question of when the most of these possibilities without removing the
the first wine will be sold in which not only enzymes charm, mystique and romance from grape cultivation
and yeast as the products of modern biotechnology are and winemaking. An equally huge challenge is the great
used in the production process, but where the grapes number of complex and interconnected obstacles, put
are also taken from a genetically modified plant. there primarily by legislation and social reticence. These
are currently blocking the commercial availability of
6.11. IN CONCLUSION transgenic grape cultivars, genetically modified yeasts
and malolactic bacterial starter cultures. It goes without
The signs are clear: recombinant DNA technology is saying that a thorough investigation of the potential
gradually gaining acceptance in the vineyard and winery. negative effects of new technologies is necessary, but
Stellenbosch in South Africa was way ahead of the game. if these hurdles cannot be removed within a reasonable
Sadly for this region, Pretorius, who is so convinced of the timeframe, both the individual consumer and the whole
many opportunities of modern biotechnology, has now international wine industry will be the loser. The previous
found employment in a wine institute in Australia. A former sections have included examples of improved grape
colleague of his from Stellenbosch, Professor Hennie van cultivars, yeast strains and bacterial starter cultures.
Vuuren, also left Stellenbosch and has meantime become Together these can make wine production more efficient
the director of the Wine Research Centre at the University and more cost-effective with a maximum profit in terms
of British Columbia in Vancouver. He was the person of product quality and a minimum consumption of raw
responsible for making and marketing a malolactic and an ingredients and damage to the environment. In short,
ethyl carbamate-free wine yeast using genetic modification should we opt for a product that is better value for money
(Pretorius, 6 August 2008, personal communication). They in all aspects if we implement modern biotechnology, or
are now being used by several wineries in North America should we stick to traditional ways on the basis of often
for commercial wine production. irrational motives?

110 Part 2: Our daily food and drink

 6.12. SOURCES Pretorius, I. S., & Bauer, F. F. (2002). Meeting the consumer
challenge through genetically customized wine-yeast
Baldwinson, J. (1975). Plonk and Superplonk. London, strains. Trends in Biotechnology, 20(10), 426-432.
Coronet Books. Pretorius, I. S., & Hoj, P. B. (2005). Grape and wine
Bisson, L. F., Waterhouse, A. L., Ebeler, S. E., Walker, M. A., biotechnology: Challenges, opportunities and potential
& Lapsley, J. T. (2002). The present and future of the benefits. Australian Journal of Grape and Wine
international wine industry. Nature, 418(6898), 696-699. Research, 11(2), 83-108.
DeFrancesco, L., & Watanabe, M. (2008). Vintage genetic Vivier, M. A., & Pretorius, I. S. (2002). Genetically
engineering. Nature Biotechnology, 26(3), 261-263. tailored grapevines for the wine industry. Trends in
Pennisi, E. (2005). Wine yeast’s surprising diversity. Science, Biotechnology, 20(11), 472-478.
309(5733), 375-376.

Chapter 6: Wine: one of the oldest biotechnological products 111

7 MEAT FROM THE BIOTECH VAT

“According to the law of Torah it is forbidden to let animals suffer.”

In Judaism the word Torah is normally used to describe the first five books of the Hebrew Bible. These Five Books
of Moses form the basis of the Jewish faith. Caring for animals is thus not only from this time as appears from this
old Jewish saying. There are many traditions and rituals involved in the slaughter of cattle for human consumption,
whereby concern for the animal’s welfare is also a priority. For example, Muslims can only eat meat that is halal, in
other words, meat that is slaughtered according to strict guidelines. One of the conditions is that the butcher should
ensure that the animal is comfortable. The Jewish religion has similar guidelines (kosher). Meat is consequently
a very traditional product, but not a traditional biotechnological product. Nevertheless, for decades meat has
undergone processes involving the use of enzymes. These processes can thus justifiably be called biotechnological.
In addition, in the production of some sausages there is a fermentation step that gives the sausage a slightly sour
taste. This fermentation is sometimes helped along with the addition of bacteria cultures. As with bread, cheese
and wine, which are really traditional biotech products, it is clear that modern biotechnology is making ever bigger
inroads into cattle breeding and the meat industry. A number of topical examples are discussed in this chapter.

BIOTECH ALSO ENTERS MEAT PRODUCTION

LOOK! SAUSAGES AT BIRTH!

J. Tramper, Y. Zhu, Modern Biotechnology – Panacea or new Pandora’s box? 113

DOI 10.3920/978-90-8686-725-7_7, © Wageningen Academic Publishers 2011
 7.1. SCOPE nutritional value which can vary greatly for chickens,
pigs and cows. The addition of enzymes to animal
It will be a while yet before we find any meat on our feed can drastically increase the availability of the
plates that comes from genetically modified (also various nutrients, resulting in possible improvements
called transgenic or recombinant) animals. It is in the efficiency of feed utilization and in positive
important, however, that we start thinking about this environmental effects. Many of these enzymes are
issue now, in the light of the many developments in made using genetically modified microorganisms.
this area. And just because it isn’t currently happening, There is an increasing likelihood that the most
doesn’t mean that modern biotechnology has not important components, in particular corn and soya,
already made some inroads into the cattle rearing will also be products of modern biotechnology. As far
and meat processing industry. On the contrary, this is as social acceptance is concerned, the use of these
presently happening in the area of animal feed, not to transgenic crops has huge consequences.
mention the injection of hormones into animals. In both
cases recombinant DNA technology is already playing Milk substitutes
a leading role. Enzymes in meat processing are a fact In animal husbandry in West Europe and North
of life now, and it is likely that some of them come from America it is common practice to separate the
genetically modified microorganisms. All these issues ‘production mother’ as soon as possible from her
are discussed in turn in this chapter. Towards the end young. The sooner a suckling piglet or calf can be
there are a few short sections on meat from cloned weaned off maternal milk, the better for the cattle
animals, other new developments and biotechnological breeder, because the mother can then quickly be
meat substitutes. fertilised again; the suckling of young animals has
a contraceptive effect. Feed made from grain and
7.2. ANIMAL FEED vegetal proteins is not usually given to young piglets,
because their stomachs and intestines are only able to
Introduction tolerate milk. However, the addition of (recombinant)
Commercial animal feed consists mainly of barley, rye enzymes (see next page) facilitates the digestion of
and wheat grains. Corn and soya are also important these vegetal proteins, enabling the pig farmer to give
components. The composition of animal feed is a cheaper blend of feed to the piglets. In some cases
such that it facilitates the most efficient uptake of it is even possible, with the addition of enzymes,
carbohydrates, proteins and fats by the cattle. The to completely replace the creamed-off milk, which
grain type and variety, the weather and in particular the is normally used as a substitute for maternal milk,
ripeness of the grain, largely determine the available without slowing down the growth of the piglets.

114 Part 2: Our daily food and drink

 Enzymes add nutritional value
LOTS OF MILK SUBSTITUTES
Still another question is whether all animal feed
ARE AVAILABLE THESE DAYS
additives should also be recombinant-free. Enzymes
THANKS TO BIOTECHNOLOGY
made using recombinant microorganisms play an
important role in the production of animal feed, just as
they do in cheese, bread and wine. These catalysts
can ensure better digestion and take-up of the
nutrients in the feed. As has been previously described,
commercial feed mixes are blended to provide cattle
COLA

with an optimum mix of starch (carbohydrates),

proteins and fats. Maize is usually the most important
source of energy in poultry diets. But if, for economic
reasons, different grain types like barley, rye or wheat
are used instead, productivity is considerably lower
than expected from the nutrient content, because
Transgenic crops digestibility is poor. Furthermore, a similar alternative
In the chapter on bread we saw how society’s demand diet often causes sanitary and health problems such
for a separation of the supply flows of genetically as sticky excrement and poor quality manure, with
modified and conventional crops is becoming all the consequences thereof (infection, soiled eggs,
increasingly urgent. In order to achieve an effective etc.). The extent of the problems is closely related
separation of the two flows they must be processed to the sort of grain, the specific cultivar, the climatic
strictly separately, and there must be proper methods conditions under which the crop was grown, and, most
of demonstrating the separation. Only then are of all, the time at which the grains were harvested (the
labels reliable. Only then can the consumer make ripeness). These problems can largely be prevented
an informed choice and be sure of whether he/she is by adding suitable enzymes, which, for example,
buying recombinant-free food or not. break down grain-cell walls, facilitating the uptake of
Truly “recombinant-free” meat obviously requires nutrients by the animals. The addition of enzymes has
that the animals from which it is sourced are fed with been occurring on some scale for the last 20 years.
crops that also bear the label “recombinant-free”. A still Recombinant DNA technology is making it increasingly
unanswered question is what that implies in terms of attractive in terms of cost, because microorganisms
cost and whether the average consumer is prepared can be optimised by genetic modification to produce
to pay that price. the required enzyme.

Chapter 7: Meat from the biotech vat 115

 Adding enzymes helps the environment happening in recent years, mainly as a result of an EC
The addition of carefully selected enzymes to ruling of May 2000. Since the end of the 1980s meat
animal feed can have a beneficial effect on the and bone meal are no longer permitted in ruminant
environment, as has been mentioned above. Better feed, because of the risk of BSE (Mad Cow Disease).
digestion of proteins, carbohydrates and fats, the On 1 January 2001 an extended ban was introduced
main components of the feed, means less and better under pressure from consumers. As a result, these
quality manure. There is also an advantage in terms of cheap sources of protein and phosphorus are no
minerals, especially phosphorus, which is an essential longer available to producers of pig and poultry feed.
element for life and development. Sixty to sixty-five In principle, then, the addition of phytases has a three-
percent of phosphorus in grains is present in the form fold positive effect. Iron, zinc and other essential metals
of phytate, which animals with only one stomach (e.g. are better taken up; no extra phosphate is needed in
chicken and pigs) find difficult to digest. This has major the feed; and less phosphorus is released into the
consequences both for the feeding of these animals environment via the manure. Tests have substantiated
and for the environment. Phytate forms complexes with the latter effect: up to 42% less phosphate in chicken
the ions of a number of important other elements such manure and up to 35% less in pig manure has been
as iron and zinc, which consequently cannot be taken observed.
up efficiently from the feed by the animals. In order to
ensure that the animals get enough phosphorus, extra LESS PHOSPHORUS IS RELEASED INTO THE ENVIRONMENT...
inorganic phosphate is added to the feed, because this ...AT NIGHT IT USED TO BE QUITE A SHOW!
can be successfully taken up, even with iron and zinc
as counter ions. Most of the phosphorus from animal
feed is, however, released into the environment via
the manure in the form of phytate and as such is one
of the biggest environmental problems of intensive
farming, i.e. exuberant algal growth suffocating lakes
and coastal waters (eutrophication).
Various microorganisms and fungi in particular, make
phytases. These are enzymes that break down phytate Yet phytase is still only added to animal feed in a
into inorganic phosphate, amongst other things. This limited number of countries. This is partly due to its
latter can be taken up successfully by animals with limited availability, since microorganisms don’t make
one stomach. It seems only sensible, therefore, to add very large quantities of them. It is therefore only
such enzymes to animal fodder. In fact this has been to be expected that some companies have tried to

116 Part 2: Our daily food and drink

use copies of the phytase gene in better production can be stored for several years. The best thing about it
organisms by genetically modifying them. DSM-Gist is is that the rapeseed can be added without any further
one of the companies that succeeded in this venture, processing to the animal feed; there is no need to
bringing a commercial product for use in animal feed remove the phytase from the seed. Chicken and swine
onto the market more than ten years ago. Since then, feed already contains a little rapeseed, so by replacing
there has been much activity in this market. An alliance normal rapeseed with this transgenic rapeseed, there
between Novozymes and Roche Vitamins Ltd was is no longer any need to add phytase. Figure 7.1 shows
set up in January 2001 to make enzymes for animal just how well the chicks thrive on it.
feed, and guaranteed these companies a strong Shortly before the turn of the last century, it was
position in this market. In 2003 DSM-Gist acquired expected that commercial introduction would soon
this vitamin concern from Roche. To prevent DSM-Gist follow. However, that has not yet happened. Probably
monopolising the area of enzymes for animal feed, in part because resistance to transgenic plants has
the EU anti-trust legislation required the company to not declined, and partly perhaps because DSM-Gist
transfer the phytase production process to BASF, with earned plenty from the recombinant microbial phytase,
whom they had a similar alliance. Today a long list of and partly too because the expression level of the
manufacturers and suppliers is available. A fact sheet enzyme in rapeseed is low. In the meantime, Mogen
on phytase has recently been published by Jacela et and Plantzyme are defunct.
al. (Jacela et al., 2010); this fact sheet can also be Week
Transgenic seeds
found on the website of the American Association of Aspergillus niger
4
Swine Veterinarians43. Phosphorus cattle food
Control seeds
Another interesting development in this area came No additives
when the Dutch company Mogen inserted the phytase
Transgenic seed
gene in the rapeseed plant by modifying the genetic Aspergillus niger
2
material. In 1997 this company set up a joint venture Phosphorus cattle food
Control seeds
with DSM-Gist called Plantzyme to commercialise No additives
these developments. The phytase was made in 0 200 400 600 800 1000 1200
Growth (grams)
the seed of the recombinant rapeseed plant. The
advantage of this is that harvesting can take place in Figure 7.1. The effect of transgenic seed containing phytase
the usual way. In addition, the enzyme in the seed is on the growth of chicks over four weeks. For comparison the
apparently so well protected from negative influences, effect of feed with added Aspergillus niger (fungus) phytase, with
that it remains active for longer. As a result the seed inorganic phosphate, with conventional non-transgenic seed and
finally feed without any of these additives (Koenderdam, 1997).
43
http://www.aasv.org/shap/issues/v18n2/v18n2p90.html

Chapter 7: Meat from the biotech vat 117

 7.3. GROWTH HORMONES result in a higher concentration of these in the milk,
with all the consequences thereof. Others worried
“Approximately 80% of all beef production in the about further increases in overproduction and the
US is made possible thanks to the use of growth consequent disappearance of the family farm. The use
hormones.” of rBST was therefore banned in Canada and the EU.
In the EU this moratorium on the use of rBST and the
In early 1996 the above statement was made in a other hormones came into force in 1988. There was
Dutch agricultural newspaper by Philip M. Sheng, also a consequent ban on the import of meat and milk
director of the US Meat Export Federation. What this products from cows injected with hormones.
quote shows is that the addition of hormones to cows
is economically advantageous. The growth hormones
GROWTH HORMONES IN THE USA
in question are sex hormones like progesterone, CHECK IT OUT!
testosterone, estradiol and synthetic hormones such ONLY ONE WEEK OLD!
as zeranol and trenbolone; in addition there is the
recombinant bovine growth hormone rBST. By injecting
rBST cows grow faster, milk yield increases, the meat
is leaner, and the feed is used more efficiently, with
less manure as a consequence. This obviously has a
very commercial appeal to cattle farmers.
The journal Genetic Engineering News (GEN) has
a column called “Point of View”. In the edition of
15 January 1998 this column was entitled ‘Public
Education Still Needed on Biotech’. It was written by
Isaac Rabino (1998), a professor of Biological and Along with insulin and the swine diarrhea vaccine,
Health Sciences at the State University of New York. rBST is one of the first commercial products of modern
His article also maintained the following: that the biotechnology and, as the previous paragraph shows,
complexity of the biotechnological discussion points it has been controversial since the late ‘80s. By the mid
is clearly illustrated by the production of rBST; and ‘90s this discussion had flared up again in the EU. The
that action has been taken against the use of rBST daily and weekly papers began to express the view
by consumer groups, who feared that udder infection that it was no longer possible to uphold the European
(mastitis) caused by increased milk production would ban on rBST and on other growth hormones allowed
lead to an increased use of antibiotics which would in the US. A new global trade agreement, signed in

118 Part 2: Our daily food and drink

1995 by the EU and the US, included a statement still no final agreement and the discussion continues
declaring that this sort of hormone product, as well as about compensation rules and possible labelling
genetically modified products, could only be banned of “normal” meat as hormone-free. It costs the EU
on the basis of scientific arguments. In that same year more than 100 million dollars a year to maintain this
an international committee of scientists, summoned recalcitrant attitude.
by the EU, concluded that meat and dairy products
MAD BUREAUCRAT’S DISEASE
from cows treated with hormones do not constitute a
risk to consumer health, provided the hormones are
administered to the cows under strictly controlled
conditions. This paved the way for the lifting of the
import ban on these products. At the end of 1995 the
United States government then appealed directly to
the WTO (World Trade Organization) to lift the ban on
imports. Conversely, the EU did all it could to prevent
the import ban from actually being removed. Earlier
arguments, such as the supposedly excessively
fast development of the skeleton with ensuing pain
(growing pains), tumor formation, reduced fertility, What is clearly undesirable is a repeat of the hypocritical
increased stress and aggression among the treated situation we saw in the 1980s in Germany concerning
animals, were all regurgitated. insulin (see Section 1.5): American farmers being
In 1998 these American meat and dairy products allowed to use hormones and therefore having an
were still not on the European market and the papers economic advantage on the European market with the
reported an escalation of the row into a full-scale war, free trade of their milk and meat products, because the
in which talc and gelatin from hormone-induced cows European farmers are not being allowed to use these
and used in medicines, were thrown into the fray (see hormones. This is aside from all the other pros and cons
also Textbox 7.1). The financial stakes were also in the matter of hormones being used to stimulate growth
blown up out of all proportion. The discussion was and milk yield in cows. In the US this feud has led to
not only to do with rBST and the other hormones an increasing debate in society on such matters.
permitted in the US. EU officials also dragged RANT One of the results of this is that on American
A
E
GU

in the danger of BSE. This was referred to as rBST supermarket shelves you can now find milk that
ED

the ‘Mad Bureaucrat’s Disease’ of Brussels in FREE is clearly labelled as ‘guaranteed rBST free’,
American papers. As far as we know there is both on the cap as well as on the shelf label.

Chapter 7: Meat from the biotech vat 119

 TEXTBOX 7.1. to ban imports of drugs containing tallow or gelatin.
Recombinant gelatin. On 8 December 1999 there was an article in the
AgraFood Biotech journal (Anonymous, 1999)
The rBST affair reveals yet another distinct difference claiming that the American firm FibroGen could
between the US and Europe, namely that of successfully produce gelatin with genetically
entrepreneurship. People in the USA are much quicker modified yeasts and plants (tobacco). According to
to turn a problem into an opportunity than those of us in the company, the main advantage was the safety
Europe. The following example is a clear demonstration of the product. Gelatin from animals carries the
of that. risk of being contaminated (with, for example,
Gelatin is made from collagen. Collagen is the protein prions that cause Mad Cow Disease), or invoking
of the fibrous connective tissue of bone, cartilage the wrong immunological reactions in vaccinations
and skin. It is the most prevalent protein in the higher against measles, mumps, scarlet fever, etc. The new
vertebrates. Collagen, and therefore gelatin too, is process enables FibroGen to make ‘customised’ safe
obtained from animal carcasses. Gelatin is regularly gelatins for specific applications. The company was
used in food, but a secondary important application extraordinarily quick to capitalise on the escalating
is in the pharmaceutical industry. It is an ingredient rBST affair. When we looked at their website44 again
found in hard capsules, soft gels, plasma expanders, in 2010, we noticed that they now offer rDNA-yeast
tablet binding agents and coatings, and vaccine gelatins that are similar to human gelatin and that
stabilisers. Therefore, it is understandable that the US are non-immunogenic, as demonstrated by research
pharmaceutical industry reacted angrily when, as a in collaboration with groups that specialise in gelatin
result of the bovine hormone affair, the EU threatened allergy.

In 2003 the biotech company Monsanto lodged had been making rBST-free claims for years. Stanley
a complaint against the dairy company Oakhurst Bennett, the director of Oakhurst Dairy45, maintained
Dairy from Portland, Maine in the US, because of that the company had been producing rBST-free milk
a misleading claim. According to Monsanto the for five years because consumers liked it: “We are
advertising done by this family company gave the in the business of marketing milk, not Monsanto’s
impression that milk that didn’t come from rBST cows drugs.”
was safer than rBST milk, while scientific studies Five years later, in 2008, the discussion flared up
demonstrated that this was not the case. What is again in the US, while in the EU it hasn’t been on
remarkable is that the biotech company waited until
this moment to take action, when the dairy company 44
www.fibrogen.com/collagen_gelatin
45
www.oakhurstdairy.com/about

120 Part 2: Our daily food and drink

the news agenda for years. There was a heated from pineapple, are now used to tenderise meat. It is
discussion about labelling, supermarkets removed simply a matter of time before enzymes made with
rBST products from the shelves and major dairy recombinant microorganisms make their entry in this
companies stopped buying and processing rBST area too. Yet modern biotechnology already has its
milk. A report published in mid-2009 spoke about “foot in the door” in meat production.
an overreaction and a storm in a teacup: only one in It is extremely important to maximise the profit on
ten consumers is worried enough to want to change commercial products of meat processing. To this
purchasing and consumption behaviour. Half of the end methods are being developed to restructure
“worriers” already buy alternative products. low-value pieces, scraps and juices to make them
look tastier and more appetising and to enhance
DID YOU KNOW THAT PINEAPPLE
the flavour and texture, to raise the market value.
IS USED TO TENDERISE MEAT?!
These processes usually consist of cutting the meat
into small pieces, then shaping and binding. For
some years now Novozymes, the Danish enzyme
manufacturer mentioned earlier, has been selling
a mixture of specific proteolytic enzymes that
dramatically improves this process. The enzyme
transglutaminase can also make a very positive
contribution in this area. This enzyme binds proteins,
peptides and amino acids together, with the result that
7.4. MEAT PROCESSING the texture is better, lysin - an essential amino acid
- is better protected against chemical conversions,
Meat can be tenderised naturally by storing it for about fats and fat-soluble substances are locked in, heat
ten days at 2° C. This slow process of converting and water-resistant films are formed, heat treatments
muscle into meat with endogenous proteases for jelly formation are unnecessary, elasticity and
(protein-digesting enzymes) and collagenases water-binding capacity are enhanced, solubility and
(endopeptidases) ensures tender meat; the downside functional properties change for the better, and the
is moisture loss and shrinkage. Research has been nutrition value increases because various proteins,
under way since 1940 to see if this “hanging” process whose composition is complementary to limiting
(“aging” in game) can be improved by using exogenous essential amino acids, are bound to each other.
enzymes. On a commercial scale proteases from In short, a whole gamut of positive effects may be
plants, for example papain from papaya and bromelain achieved.

Chapter 7: Meat from the biotech vat 121

 Until recently the only commercially available enzyme Ajinomoto brought a transglutaminase from a
with this mechanism was transglutaminase isolated microbial source onto the market. Its use in food
from the livers of Guinea pigs. The scarcity, and the production is, however, banned in EU countries.
difficult and laborious method of obtaining the enzyme These microbial sources have highlighted cheaper
in a workable form, made it extremely expensive - production methods, especially if recombinant DNA
too expensive for use in industrial meat processing. technology is used, which is only a matter of time.
New sources are now available. For instance, This will open the way to efficient and profitable meat
transglutaminases are found in microorganisms processing, and much more. Table 7.1 shows a list
such as Streptoverticillium and Streptomyces of a great many other interesting possibilities for
strains. Several years ago the Japanese company food processing.

Table 7.1. Summary of application possibilities of the microbial enzyme transglutaminase in food processing.

Source Product Effect

Meat Hamburgers, meatballs, dumplings, Better elasticity, texture, taste and
shaomai Tinned meat Frozen meat aroma. Good texture and appearance.
Compressed meat Improved texture and lower costs.
Restructuring of meat
Fish Fish pie Improved texture and appearance
Krill Krill pie Improved texture
Collagen Shark fin imitation Imitation of tasty food
Grain Baked food Improved texture with more volume
Soya beans Mapuo-Doufu Baked Tofu (Aburaage) Improved shelf life. Improved texture
Tofu Improved shelf life
Fruit and vegetables Celery Food preservation
Casein Stimulators of mineral absorption Cross- Improved mineral absorption in the
linked proteins intestine. Reduced allergen reactions
Gelatin Sweet food Food low in calories and good texture,
form and elasticity
Fat, oil and proteins Hard fats Lard substitute with good taste, texture
and aroma
Vegetable proteins Protein powders Gel formation with good texture and
taste
Herbs Herbs Improved taste and aroma

122 Part 2: Our daily food and drink

 7.5. CLONED MEAT everything that is edible. Sooner or later we will be
confronted with the question as to whether meat from
In Chapters 1 and 2 genetically modified, so-called cloned animals is suitable for consumption. In fact we
transgenic animals, and their clones, were introduced. have already reached that point. A cattle company in
We will come back to this topic again in Chapter 14. Canada already applied in 2003 to the authorities to
The number of transgenic and cloned animals is still be allowed to sell meat from cloned cattle for human
relatively small, but the number is rapidly rising. Even consumption.
in a country like New Zealand, with campaigns against A more recent development began on 28 December
transgenic animals (see Section 2.2), the authorities 2006 when the Centre for Veterinary Medicine (CVM)
granted permission in April 2010 to the country’s of the FDA published a three-part discussion document
leading agricultural research company to continue its entitled: A Risk-Based Approach to Evaluate Animal
work on genetically modified livestock . This decision
46
Clones and Their Progeny – Draft. It consisted of a draft
means the animals, which include a herd of 100 risk analysis, a proposal for a risk management plan
genetically modified (GM) cows, can be returned to an and draft guidelines for the industry. Before the draft
active breeding program. Not everyone is happy with risk analysis was published, it had already been looked
this decision. “We are appalled,” said Claire Bleakley at by independent scientific experts, who agreed with
of GE free NZ47. “They now have carte blanche to the methods the CVM used to evaluate the data, and
produce any number of GM animals with no way to therefore supported the conclusions.
properly assess the potential danger to health and According to the CVM meat and milk from cloned cows,
the environment, and the controls are no stricter than pigs and goats are just as safe to eat as conventional
those for previous decisions.” food and thus require no special labelling. The
According to Jonathan Cowie (2000) there will be a American Biotechnology Industry Organization (BIO)
major global food crisis in the middle of this century. backed the CVM in this matter and said in a press
He bases this conclusion on the report “GM crops: The release that the results are consistent with numerous
Social and Ethical Issues” of 1998, which was drawn studies already showing that food from animal clones
up by a working group (of which Cowie was a member) and their offspring is safe. The CVM invited with the
of the English Society of Biology and six affiliated 2006 draft the American citizens to give their opinion
associations (whose specialist interests range from on the document. BIO had this to say:
agricultural production to ecological conservation).
Such a food crisis will mean the inevitable use of “While there are currently no products from cloned
animals and their offspring in the market, the
46
www.abc.net.au/science/articles/2010/04/15/2873674.htm publication of the FDA’s draft risk assessment
47
www.gefree.org.nz

Chapter 7: Meat from the biotech vat 123

 will begin an essential public discussion on the The article quotes surveys showing that the average
technology and how it can be successfully used by American is very concerned about the introduction of
farmers and ranchers.” meat and milk from clones in the supermarket, and
that members of Congress are preparing legislation
The American Meat Institute responded by issuing a requiring food from clones and their offspring to be
warning that the FDA should be extremely cautious labelled as such. According to one of their reporters
about allowing such animal clone products on the there is also a threat of a new controversy about
market because many consumers have difficulty whether food manufacturers can label their products
accepting this technology. They urge the government from the offspring of cloned animals as ‘organic’, if the
not just to uphold the safety of these products in farmers concerned comply with the requisite federal
the political arena, but also to make it easier for the criteria. It is further stated that the debate about cloned
consumer to obtain a better understanding of what food has hotted-up and that in 2008 there might well
cloning actually entails, so that the overall confidence be a battle between the cloning industry, the anti-
of the consumer in food provision is maintained. cloning supporters, the FDA and Congress. The article
emphasises that the public should not overestimate
CONSUMERS HAVE DIFFICULTIES the differences between cloning cattle to improve
ACCEPTING BIOTECH FOOD the breed, and what current-day cattle breeders are
AND WHY DON’T YOU TRUST now doing to improve their herds. In conclusion, it is
YOUR BUTCHER ANYMORE? remarked that the facts are less threatening than many
Americans believe.
On 15 January 2008 the CVM-FDA published a three-part
final report of almost 1000 pages, the main conclusion of
which was: Milk and meat from healthy clones and their
offspring are just as safe for consumption as that from
normal cows, pigs and goats. The scientific committee
of the EFSA simultaneously brought out a draft opinion
in support of this conclusion. The European Group on
Ethics in Science and New Technologies (EGE48) took a
Proof that the meat institute’s comment is not entirely fiercely opposing point of view, saying that the cloning of
wide of the mark appeared in an article in the Washington animals for food production is not justified for ethical and
Post of 10 February 2007 (Anonymous, 2007). The welfare reasons. And so began the commotion about
headline runs as follows: “Frankenfood? Not quite.”
48
ec.europa.eu/european_group_ethics/index_en.htm

124 Part 2: Our daily food and drink

this in the EU. In mid-2009 a complete ban on cloned “One biologist practises his profession with a pair of
meat in the EU seemed imminent. Even the then newly binoculars, the other with test tubes. Midas Dekkers
appointed MEPs seemed to have similar views to the (1946) does it with a typewriter, microphone or camera.
EGE about it. A year later on 7 July, the day of the final His interest in animals grew from his time studying in
revision of this chapter, the New York Times headed an Amsterdam, his interest in people was awakened in
article with: Europe Seeks to Ban Food from Clones. his parents’ cafe. Midas Dekkers writes mostly about
James Kanter, the reporter, opened with: “The European the common ground between both. Since this is rather
Parliament asked on Wednesday for a ban on the sale remarkable, it is often rather amusing. Before you
of foods from cloned animals and their offspring, the know it, you’ve really learned something.”
latest sign of deepening concern in the European Union
about the safety and ethics of new food technologies.” HEALTHY EATING IS ABOUT AS
What is clear to us is that not everything in the area of FEASIBLE AS A HEALTHY LAMPPOST
modern biotechnology is accepted unquestioningly by
society, neither in the EU nor in the US. It might once
have been the case for the US public, but we now see
the American government itself encouraging a serious
SURE!
public debate about this thorny issue - something that
is not really happening yet here in the Netherlands or
anywhere else. Times change.

7.6. NEW DEVELOPMENTS

“Healthy eating is about as feasible as a healthy

lamppost.”

On Monday 25 June 2001 the Dutch public debate

“Biotechnology and Food” began with the opening
event “Food and Genes” (see also Chapter 8). At
some point during this day there was an appearance
by Midas Dekkers. His appearance was announced in What we remember are his statements about meat.
the program booklet with the above quote. The booklet He noticed that most people are very opposed to
described him as follows: the thought that a piece of steak, or meat in general,

Chapter 7: Meat from the biotech vat 125

 could be made entirely within a factory. Remarkable, in about six years. But cultivating a real piece of meat
he says, because practically all our food is made for would take at least twice as long. Sebastiaan Donders
the most part in a factory. Now, however, industry is nicely illustrated this research process in the article and
working on a recipe for minced meat from a lab. Figure 7.2 is a loose interpretation of it. In the article a
few other curious studies in this area are also mentioned
Test-tube steak (Textbox 7.2).
“Researchers in the Netherlands have created
enzyme transfer
what was described as soggy pork and are now treatment on medium

investigating ways to improve the muscle tissue in

the hope that people will one day want to eat it. No young skeleton isolated stem cells
muscle stem cells in culture
one has yet tasted the product, but it is believed the emerging
artificial meat could be on sale within five years.” stem cell clones
placing on matrix differentiation
and starvation to muscle cells
These statements are made by Nick Britten in the
cells fuse
Telegraph of 29 November 200949. Four years earlier emerging muscle
meat from into fibers and cultivated
eventually cells from stem
the headline to a full-page article by Annemarie Eek the lab
into meat cell clones
stem cell clones

in a Dutch biology journal (Eek, 2005) had a more

Figure 7.2. Steps in the development of meat from the lab,
cautious message: “It sounds like science fiction, but
adapted from Eek (2005).
Dutch researchers are cultivating meat from pig stem
cells. Due to practical problems it will still be a while Vegetarian groups and animal rights campaigners see no
before cultivated meat appears in our supermarket.” ethical objection if meat was not a piece of a dead animal,
She wrote that the Netherlands was the first country to says Nick Britten in the Telegraph article. Meat produced
systematically conduct research into the development in the lab could also reduce greenhouse gas emissions
of cultivated meat. The idea is that stem cells (see associated with real animals, according to him. However,
also Chapter 14) multiply and then differentiate into the Vegetarian Society said: “The big question is how
muscle cells, which then fuse into muscle fibres. These could you guarantee you were eating artificial flesh rather
fibres would finally form a real piece of meat together than flesh from an animal that had been slaughtered. It
with connective tissue and fat cells. There’s still a lot would be very difficult to label and identify in a way that
of research to be done, however, before this stage is people would trust.” A week earlier Prince Charles, a
reached. The expectations then were that it would be fierce opponent of GM food, warned that people were
possible to make a sort of minced meat from the fibres creating problems by treating food as an easy commodity

49
www.telegraph.co.uk/science/science-news/6680989/Meat-grown-in-laboratory-in-world-first.html

126 Part 2: Our daily food and drink

TEXTBOX 7.2. stem cells from sheep, but pieces of frog muscle grew
‘Happy Birthday’. a tiny bit. The minute pieces of frogs’ legs, in Calvados
sauce, surrounded by a host of live frogs, were
Back in 1912 the Nobel Prize winner and surgeon presented at a bio-art exhibition in Nantes in March
Alexis Carrel succeeded in keeping a piece of heart 2003.
muscle from a chicken embryo alive. Every two days A more serious attempt to develop cultivated meat
the tiny muscle received fresh food and a clean bottle. was made by tissue engineers at Touro College in
The story goes that Carrel sang ‘Happy Birthday’ to his New York at the behest of NASA. In order to be able
little muscle every year and that it wasn’t until 32 years to provide meat for astronauts in space, they cultivated
later that the piece of meat died, along with its carer. goldfish tissue in a Petri dish. Unfortunately, the meat
In West Australia artists and tissue engineers only grew on serum from calf foetuses, which meant
collaborated on cultivating meat. It didn’t work with that still more animals were needed for its production.

rather than a precious gift from nature. The real problem consumers still doubted the safety of Tropina, fearing
is in our opinion the answer to the question “how are we that it contained aromatic hydrocarbons. Opposition in
going to feed the world in 2050 in a sustainable way?” Japan even led to a complete ban on edible proteins
produced by the petrochemical industry. Governments
7.7. BIOTECHNOLOGICAL MEAT SUBSTITUTES around Europe demanded more research. Further
studies showed that the product was not carcinogenic.
There is an urgent need for new sources of protein On these grounds Tropina was permitted, albeit in
as an alternative to meat. This is because of the restricted quantities and only for export! However,
growing world population and increased affluence when the price of oil began to rise, the substrate
in countries such as China and India. Back in the on which the yeasts were cultivated became quite
1950s oil companies, and not the food industry, had expensive. BP therefore decided to stop production
already begun producing microbial protein as a meat of Tropina, because it could no longer compete with
substitute, the so-called single cell protein. They used soya protein (Israelidis, 1988). The British company
by-products as a substrate for cultivating certain types ICI ran into the same problems. It introduced Pruteen,
of microorganisms. British Petroleum (BP) brought a protein made from bacteria grown on methanol as
Tropina onto the market, a protein originating from feed. The raw protein content of Pruteen was 72% and
a yeast that was grown on alkanes. BP first studied it was brought onto the market as a high-value protein.
Tropina for twelve years for toxicity and carcinogenic In other words, it had a well-balanced amino acid
properties, but found no proof of harmful effects. But composition. ICI was building a Pruteen factory with

Chapter 7: Meat from the biotech vat 127

 60,000 ton capacity per year when the company was that of whole insects still needs to be investigated. In
confronted with the oil price rise. Despite successful addition to these technological aspects it is vital to find
engineering, it turned out that Pruteen too could no out whether consumers will accept food made from
longer compete with proteins from soya and fish. insect cells and under what conditions, and whether
A microbial protein that enjoyed more success is this can be influenced by a suitable marketing strategy
mycoprotein. It was produced with a Fusarium fungus (Verkerk, Tramper, Van Trijp, & Martens, 2007).
and processed into Quorn products. In 1986 Quorn was
launched on the British market as a meat substitute. BIOTECHNOLOGICAL MEAT SUBSTITUTES
These products satisfy a number of important
ONE ‘BIG HOPPER’ MENU PLEASE!
consumer needs. For example, they are healthy, easy
to prepare and have the same flavour and texture as
normal food. Quorn is now seen as more of a meat
substitute than, for example, soya protein.
A completely different and interesting group of
organisms that can be used as a source of protein are
insects. In the Western world this is virtually unheard
of as a food group, but it is becoming increasingly
important. Edible insects (more than 1,380 types)
have long been accepted as a nutritious source of
proteins, vitamins and energy in many non-Western
countries. Not all varieties are suitable, however, for
large-scale breeding, because they are susceptible
to disease. Modern biotechnology does, however,
offer an alternative in the form of insect cell cultures, 7.8. IN CONCLUSION: HAPPY MEAT!
whether or not they are infected with a (recombinant)
virus. Insect cells can be grown in bioreactors on a In 2006 the Dutch VPRO science programme NWTV
large scale under controlled, closed conditions. There made an online appeal inviting people to come up
are also several possible ways of purposely and with a new name for cultivated meat50, meat-like
rationally changing the composition of insect cells, material thus that is made from muscle cells grown
thereby changing the nutritional value. Whether this in bioreactors. Three of the more than 200 entries
is feasible on a technological level and whether the were rewarded by the jury with an internet radio. The
nutritional value of insect cells is comparable with
50
http://noorderlicht.vpro.nl/artikelen/28570228/

128 Part 2: Our daily food and drink

winners were La Box from Joop de Meij (a vegetarian), Note added in proof:
Ewart Kuijk with Kreas (Greek for meat) and Dafne The British food authority FSA reported in August 2010
Westerhof from pork paradise The Promised PigLand that meat from a cloned bull had ended up in the food
with Happy Meat: chain. Moreover milk of offspring of a cloned cow has
also reached consumers.
“Then you know you’re going to eat a tasty meal
and no animal had to suffer for it.”

Isn’t that what we all want?

Chapter 7: Meat from the biotech vat 129

 7.9. SOURCES Jacela, J., DeRouchey, J., Tokach, M., Goodband, R.,
Nelssen, J., Renter, D., & Dritz, S. (2010). Feed
Anonymous. (1999, 8 December). Gelatin to be manufactured additives for swine: Fact sheets–high dietary levels of
recombinantly. AgraFood Biotech, p. 28. copper and zinc for young pigs, and phytase. Journal of
Anonymous. (2007, 10 February). Frankenfood? Not quite. Swine Health and Production, 18(2), 87-91.
Washington Post. Koenderdam, I. (1997, 18 January). Eerste product Mogen
Cowie, J. (2000). Genetic modification and the meat market. nadert de markt. Chemisch Weekblad, p. 1.
Nature, 404(6781), 921-922. Rabino, I. (1998, 15 January). Public Education Still Needed
Eek, A. (2005, 30 September). Biefstuk uit het buisje. Bionieuws. on Biotech. Genetic Engineering News.
Israelidis, C. J. (1988). Nutrition – single cell protein, Verkerk, M., Tramper, J., Van Trijp, J., & Martens, D. (2007).
twenty years later, Biopolitics: Proceedings First Insect cells for human food. Biotechnology Advances,
Biointernational Conference (Vol. 1). 25(2), 198-202.

130 Part 2: Our daily food and drink

8 “FRANKENFOOD”

“My concern is if we don’t have a broadly educated public … charlatans out there will be able to play on public fears.”

This is a quote from Donna Shalala, a professor of Political Sciences at the University of Miami, where she has also
held the post of President since 2001. Donna Shalala is considered to be one of America’s best leaders51 and is not
afraid of controversy, as demonstrated by the above forthright remark she made during a lecture on gene technology for
scientists. Unfortunately her fears are not unfounded and the concerns many people have about genetically modified
(transgenic) food crops are largely based on misleading information. The objectionable term Frankenfood, used by
opponents to describe food made from what they call genetically manipulated plants, conjures up negative associations.
The size of the rift between biotechnologists and the anti-GM food lobby and the extent of the unwillingness of either
to reach out to the other is plain to see.

THERE’S A GREAT GAP BETWEEN

In this chapter we will look at a number of topics related to THE BIOTECH AND ANTI-GM LOBBY
genetically modified food. The first section will deal with our food
production and more specifically “GM foods”. Secondly we will I’M NOT CLOSING IT!
look at whether the concerns about GM foods are justified. In the IF YOU WON’T, I WON’T!
third section we will consider whether GM foods are harmful to
our health. After this section we will see that it is not just
consumers but also farmers who are concerned about
the production of GM foods. In the fifth section we will ask
who is telling the truth in the debate about Frankenfood. In
the last section of this chapter we take a look at the future
of these crops. The idea behind this chapter is to present
readers with enough information so that they can form their
own considered opinion on this subject.

51
www.usnews.com/usnews/news/articles/051022/22shalala.htm

J. Tramper, Y. Zhu, Modern Biotechnology – Panacea or new Pandora’s box? 131

DOI 10.3920/978-90-8686-725-7_8, © Wageningen Academic Publishers 2011
 8.1. FOOD AND GENES Consumer Product Safety Authority (VWA) was
founded by the government with the following
The science on which gene technology is based is mission53: “The Food and Consumer Product Safety
not exactly straightforward, and yet it should still be Authority works on safe and healthy food, safe
possible for laymen to make reasonable choices based products and healthy animals. To this end, the VWA
on good, objective information, each according to their looks at the risks, evaluates them, communicates
own insight, beliefs or principles, and not on fear. In the about them and makes them manageable within
media a lot of time and space has always been devoted society.” An ambitious, heavy and difficult task, which
to this subject, but consistent information of any real from a structural point of view requires a lot of money.
scope aimed at the general public only appeared in the By 2008 budget cuts had already prevented the VWA
Netherlands in 2001, when the government set up the from being able to function as required. On 20 June
temporary Terlouw Committee. 2008 a Belgian agricultural expert wrote a very plain-
Jan Terlouw, the chairman of the committee, is speaking report about it, declaring that the VWA was
a physicist, writer and politician. Even after his not functioning properly and was being patronised by
retirement in 1996 he remained very active and is the Ministry of Agriculture. Janneke Snijder, Member
still a great advocate of human and animal rights. of the Dutch House of Representatives for the liberal
This Biotechnology and Food Committee, also party, concluded that the House of Representatives and
called the Food and Genes Committee, carried out a the Agriculture Minister were heading for a showdown
survey of what the public in the Netherlands thought about the humiliatingly bad organisational structure of
about the use of biotechnology. It also attempted to the VWA. The majority of the House wanted the whole
make accurate information accessible to a broader situation to be sorted out, before merging the VWA,
audience. In its report (2002, in Dutch) the committee
52
according to the most recent plan, with the General
maintained that there was no question about the safety Inspectorate and the Plant Disease Service (these
of food prepared using modern biotechnology and services are both part of the Ministry of Agriculture).
that the majority of the public thought this technology The Minister states conversely that the merger should
should be allowed to develop further. The report made just go ahead because “it will be years” before the
several important recommendations including better situation at the VWA improves. It seems to us there is
information, freedom of choice for the consumer and little hope therefore of the VWA being able to complete
the setting up of an independent Food Authority. its mission in the near future and to play a leading role
Shortly thereafter, on 10 July 2002, the Food and in removing irrational fears about genetically modified

52
www.voedingscentrum.nl/resources2008/eindrapport_terlouwpdf.pdf
53
www.vwa.nl

132 Part 2: Our daily food and drink

food; a situation quite typical for the EU as a whole. PRINCE CHARLES: ‘MANIPULATION OF GENETIC
At the time of the Terlouw Committee, in 2001, a MATERIAL SHOULD ONLY BE THE WORK OF GOD’
comprehensive report appeared on the Internet about
this controversial topic (A Report on Genetically I CAN’T FIND THE TERMS ‘BIOTECHNOLOGY OR GM’
Engineered Crops). The author, Charles M. Rader, is IN ANY OLD RELIGIOUS BOOK!
himself an outsider in this area. Using a great many
verifiable facts he made a careful analysis of the
various aspects and put it online in layman’s terms.
We gratefully made use of this report to put together
the first version of this chapter several years ago. For
this final version we have again, though to a lesser
extent, used the January 2008 revised version of the
Rader report. We have now, for the revision, especially into consideration other, far-reaching, consequences.
used information from a dozen or so publications Leviticus 19:19 forbids cross-fertilisation, i.e. sexual
that appeared recently in leading scientific journals. reproduction of two different plants or animals (species
However, it is still worth visiting Rader’s website .
54
or cultivars). At this point it is worth mentioning that
natural cross-fertilisation between different species is
8.2. JUSTIFIED FEARS? not at all rare, for example, cross-fertilisation in nature
between different types of birds is a known occurrence.
“Ye shall keep my statutes. Thou shalt not let thy If improvements made possible by genetic modification
cattle gender with a diverse kind: thou shalt not sow using recombinant DNA technology fall under this ban,
thy field with mingled seed: neither shall a garment then strictly speaking many other, long-used breeding
mingled of linen and woollen come upon thee.” techniques must also be excluded. Obviously it should
be possible for consumers to decide whether or not
Leviticus 19:19, King James version. they want to buy such ‘cross-hybridised’ products.
Vegetarians must be able to choose food that is free
Some people, among them the UK’s Prince Charles, of animal genes and Muslims and Jews should be
believe that the manipulation of genetic material should able to buy food free of pork and other forbidden meat
only be the work of God or Mother Nature. We respect products, even in terms of genes.
this point of view, but we don’t think it can be the basis In order to make these choices accurate labelling is
for an argument to ban gene technology, without taking vital, as also recommended in the Terlouw Committee’s
report. Since this committee was set up, far-reaching
54
members.tripod.com/c_rader0/gemod.htm

Chapter 8: “Frankenfood” 133

 legislation, discussed in Chapters 2 and 5, has been genetically modified the soya plant with a gene coding
introduced both at a national and EU level. For many for a protein rich in methionine. The gene came from
people, though, there is still the question of how certain a Brazilian nut to which some people are allergic.
we can be that genetically modified food is not harmful Tests have shown that the enriched protein from the
to our health, in the short or long term. There is also transgenic soya also causes allergic reactions in
some doubt as to whether transgenic crops will solve these people. The transferred gene therefore seems
the world’s food shortage problem and will result in to code for the protein that causes the allergy. For
healthier and more complete food, and whether they this reason the project was suspended long before
are damaging for the environment in the long term. there was any talk of marketing or consumption. This
The precautionary principle (O’Riordan & Cameron, example therefore shows why testing is necessary,
1994) is central to environmental policy (see Textbox that it happens and that it safeguards the consumer.
3.3 in Chapter 3). If we approach food safety from the Ironically, opponents of gene technology have a
precautionary principle, then all significantly modified different take on this example: “This just goes to show
crops must be carefully evaluated. This would apply not what can go wrong.” What is clear is that (products of)
just to crops that have been modified with recombinant transgenic crops should be and are more stringently
DNA technology. In contrast to the more traditional tested and regulated than ‘normal’ crops, especially
breeders, molecular biologists that genetically modify in the EU (S. H. Morris, 2007). To date the safeguards
plants using recombinant DNA technology have a seem to be more than adequate: there are no
pretty good idea what they are doing. In other words, indications that anyone has been exposed to unsafe
genetic modification is much more of a targeted GM foods in the more than ten years during which
process than traditional breeding, which relies largely they have been consumed on a large scale, especially
upon chance. Unexpected changes can still occur with in the US. That’s not something you can always say
genetic modification, but from a scientific perspective about “normal” food.
there is no reason why GM foods should be more
thoroughly tested than other new food, except with a 8.3. ARE GM FOODS HARMFUL TO HEALTH?
view to removing public concern.
The following example demonstrates the need for, As mentioned above, in 2002 the Terlouw Committee
and efficacy of, testing. Soya is a rich but low-quality maintained that there should be no doubt about the
source of protein compared to animal proteins safety of food prepared using modern biotechnology.
because it contains very little methionine - one of the According to information from 2002 (20 questions
essential amino acids. Researchers have therefore on genetically modified (GM) Foods)55, from the

55
www.who.int/foodsafety/publications/biotech/20questions/en/index.html

134 Part 2: Our daily food and drink

TEXTBOX 8.1. requires detailed characteristics of:
Risk assessment of GMOs.
• the genetically modified plant and its use as food;
In 2003, the Codex Alimentarius Committee, under the • the origin of the gene;
auspices of the FAO and WHO, adopted guidelines to • the inserted DNA and flanking DNA at the place of
harmonise the premarket process of risk assessment insertion in the genome;
of genetically modified plants intended for the global • the substances expressed (for example, proteins
market. These guidelines are intended to help individual and any new metabolic products that are the result of
countries draw up consistent legislation that provides for the new gene product);
a strong food safety evaluation process, while avoiding • the possible toxicity and anti-nutritional properties of
trade barriers. Every new gene crop has to undergo a new proteins or metabolic products;
premarket safety assessment to evaluate intentional • the protein expressed by the new gene compared to
and unintentional changes for any adverse effects that which is known to cause Coeliac’s, if the DNA
on human health, which are caused by introducing comes from wheat, barley, rye, oat or related grains;
recombinant DNA (genes). The aim is to identify hazards • the protein expressed by the new gene with regard
and, if any are found, to demand a risk assessment, to possible allergy;
and if necessary a risk management strategy, for • key endogenic nutrients and anti-nutrients including
example, non-approval, approval but with labelling and/ toxins and allergens for possible increases for
or monitoring required, or approval without restriction. specific host plants (the DNA recipients).
The process is based on science and requires the use of
methods and criteria which are proven to be predictive. Some steps of the risk assessment require a scientific
New methods must be validated and it must be shown assessment of existing information; others require
that they improve the risk assessment. experiments, in which case validation of the analysis,
The framework for evaluating potential safety risks sensitivity and verifiable documentation is required.

World Health Organization (WHO), all genetically separately, and subjected to individual safety tests,
modified food products that are now available on the because different genes are inserted in different ways
international market, have undergone a risk analysis in different GM crops. It is therefore impossible to
by national authorities in compliance with the Codex make a generally applicable declaration concerning
guidelines (Textbox 8.1). These analyses have shown the safety of GM foods. In general terms, the safety
no proof of any risk to consumer health. assessment of GM foods should, according to WHO,
WHO indicates that every GM food must be tested investigate:

Chapter 8: “Frankenfood” 135

 • toxicity; biotechnology companies involved in commercial
• allergenicity; GM foods?” This question arises from his analysis of
• specific components thought to have nutritional or the surprisingly limited number of scientific articles
toxic properties; concerning human and animal toxicological and health
• stability of the inserted gene; risk studies on GM crops, such as potatoes, corn, soya
• nutritional effects associated with the genetic beans and rice. In addition, it appears that, of the few
modification; and scientific publications available, most do not originate
• any unintended effects which could result from the from companies that grow transgenic crops or make
gene insertion. products from them. Domingo wonders how that is
possible, given that the debate on the safety of GM
A major problem when conducting safety assessments foods is causing such controversy. His overview shows
on GM foods is the use of the concept of substantial that these are mainly short-term studies, in which
equivalence. This concept is based on the principle that nutritional values rather than toxicological aspects are
“if a new food is found to be substantially equivalent examined. He concludes that long-term studies are
in composition and nutritional characteristics to an urgently required, where attention is devoted primarily
existing food, it can be regarded as being as safe as to (1) people with abnormal digestion as a result of
the conventional food.” The problem is obviously the chronic gastrointestinal disease, and (2) undesirable
definition of the word ‘equivalent’. Kuiper et al. (2002) DNA transfer into mucosa and intestinal flora.
state that this concept is not a safety assessment per According to the European Food Safety Authority
se, but does enable us to identify possible differences (Anonymous, 2008a; EFSA, 2008) (see also Chapters
between existing food and a new product. Extra 2 & 3), 90-day food trials with rodents, mainly rats, are
attention can then be focused on the differences generally sufficient to show that GM foods are safe,
in terms of the toxicological aspects. It is more of a provided that they are conducted in compliance with
starting point than an endpoint. international guidelines. This was the conclusion of the
Thus, safety assessments are conducted on all the EFSA in a report published at the beginning of March 2008
GM foods brought onto the market. To date there has in the scientific journal Food and Chemical Toxicology
been no evidence that eating such food carries any (van Haver et al., 2008). It was stated, however, that
risk for our health. Yet Domingo (2007) in his critical these trials are not suitable for identifying potential
literature review of toxicity studies on genetically allergens. The report calls for a more uniform approach
modified plants concludes with the question: “Where to food testing and the use of new profiling technologies.
is the scientific evidence showing that GM plants/ It rejects monitoring following a product’s entry on the
foods are toxicologically safe as assumed by the market; in any case it should in no circumstances be a

136 Part 2: Our daily food and drink

substitute for a solid preliminary risk analysis. corresponding conventional crops.
The first 2008 issue of Nature Biotechnology (Goodman • Four allergy tests currently used have no thorough
et al., 2008) contains an article by seven allergy experts, scientific basis.
entitled “Allergenicity assessment of genetically modified • Recent research on GM crops has shown the
crops – what makes sense?” The seven maintain the misleading nature of these four tests.
following:
They conclude that the current safety assessments
• GM crops offer major opportunities for improving according to Codex guidelines (Textbox 8.1) are based
food quality, increasing harvest yields and reducing on the currently available knowledge on food allergens
dependence on some pesticides. and risk, and that they are therefore suitable for assessing
• Before they are brought onto the market, they the potentially increased risk of allergy from a gene crop
should first be subject to a very stringent safety test, compared to the corresponding conventional crop.
as well as a detailed analysis of the allergenic risks. However, making the four non-scientifically validated
• There is (still) no documented evidence that the allergy tests compulsory may lead to the rejection of
current approved commercially grown GM crops safe and useful products, astronomical costs and to a
caused allergic reactions as a result of an allergenic possible cessation in trading without a corresponding
protein, coded by the gene inserted by genetic reduction in risk. Worse still, the use of unsuitable tests
modification. such as unvalidated animal models instead of highly
• Neither do the current commercial GM crops suitable tests could even lead to the introduction of a
have any biologically significant increased product that does contain a significant risk for a group
levels of endogenous allergens compared to the of consumers with allergies. Textbox 8.2 shows that the

TEXTBOX 8.2. every day for 26 weeks. At the end of the study period,
Genetically modified rice fights allergies. the test animals did not show any health problems, in an
initial demonstration that the allergy-fighting rice may be
What if the food we ate fought allergies instead of safe for consumption, the researchers say.
causing them? A new form of GM rice can, researchers
RICE CAN HELP AGAINST ALLERGIES
announced in 2009 (Domon et al., 2009). The new
transgenic rice designed to fight common pollen allergy
appears safe in animal studies. In laboratory studies the
researchers fed a steamed version of the transgenic rice
and a non-transgenic version to a group of monkeys

Chapter 8: “Frankenfood” 137

 world can also turn the other way around. place in a context and be compared with alternatives.
The reality of more than a decade of consuming GM Yet many recombinant DNA risk analysis protocols
foods has demonstrated that those brought onto the only concern the assessments of GMOs, i.e. not in a
market to date have not been a direct cause of harm real-life context. It should be noted here that nothing
to our health and that the safety assessments have is ‘absolutely’ safe, because (1) science can never
therefore, so far, worked as intended. Yet the scientific prove that a product will never cause harm, and (2)
basis of some tests is still shaky, as suggested above. everything comes with a certain degree of risk.
Alan McHughen, a professor at the University of 5. An analysis conducted with technical expertise is not
California and former chairman of the International enough to give solid scientific backing to the work as
Society for Biosafety Research, sums up the “fatal a whole; the reason for conducting the analysis at all,
shortcomings” of GM foods legislation in the US in should also be scientifically valid.
a letter published in 2007 in Nature Biotechnology 6. In the case of risk assessments most GMO data
(Anonymous, 2007; McHughen, 2007): requirements are excessive, in any case far beyond
the point that is necessary to be able to determine
1. Many legal bodies are still under the false impression relative safety.
that the process of transgenesis (modification with 7. Political, ethical and economic factors play a role
recombinant DNA technology) is inherently risky and in many risk assessments, thereby blurring the
that all products developed using recombinant DNA scientific focus.
technology must therefore categorically be critically
investigated. At the same time, they suppose that In fact we can conclude that (1) eating genetically
‘conventional’ breeding processes, including the modified food has thus far caused no direct harm to
genome-disturbing methods such as radiation health, (2) from a scientific point of view there is no
mutagenesis, are risk-free and therefore do not justification for testing GM foods more thoroughly than
require much or any legal investigation. other new foods, (3) further scientific justification of risk
2. Another misconception is that the combination of analyses of food is required in general, and (4) there
genes of different species is unnatural and risky. This should be more worldwide uniformity in risk analyses.
causes unnecessary anxiety among consumers and Here’s a delicate question we can ask ourselves:
is the reason behind the request for exclusive GMO would I dare eat GM foods and give it to my children?
legislation. The answer is yes, and moreover, we are undoubtedly
3. Products of a comparable risk level should undergo a doing so already! Complete separation of conventional
comparable critical examination. and transgenic animal feed, just to give an example,
4. Because risk is relative, risk assessments should take is certainly not yet watertight and is a relatively

138 Part 2: Our daily food and drink

expensive procedure, so most of our animal products erecting barriers (co-existence). EU policy-makers in all
come from cattle that have been fed some GM foods. member states are struggling with the implementation
Statements from representatives of the grain trade of coherent co-existence regulations. Demont & Devos
and the cattle feed, food and biotechnology industries (2008) appeal for flexible co-existence regulation
corroborate this. They point to the fact that the growing which explicitly takes account of economic motives.
proportion of GM corn and soya in Argentina, Brazil NB: Organic agriculture does not by definition produce
and the US makes it increasingly difficult to separate healthier products than conventional and transgenic
the normal, conventionally cultivated corn and soya crops. In a report published in 2007 by Andrew
from GM corn and soya. It is also becoming more and Staehelin and David Christopher of the American
more difficult to keep GM crops permitted by these Council on Science and Health, it was shown on the
countries, but not authorised by the EU, from entering basis of yet more scientific publications that GM crops
the export channels. This results in higher costs for and food from those crops is even safer and more
those GM-free products which the European market healthy than “organic” food56.
is demanding. The industry and trade is therefore
appealing for a more lenient European authorisation ORGANIC FOOD ISN’T NECESSARILY
policy. The downside is that a considerable number of SAFER THAN GM FOOD
consumers in the EU and US still have doubts about the
quality of GM foods. Environmental and development
organisations therefore continue to mount serious
campaigns against the cultivation of these gene crops.

8.4. MORE ANXIETY!

There is further concern from organic farmers, most

of whom think that transgenic crops are not organic,
even if they are more pest and disease resistant, and
thus require less pesticide. They are afraid that their
crops will be “contaminated” by cross-pollination with
the genes from the transgenic crops. They believe Another concern among organic farmers is an
they are entitled to protection against this, in a passive accelerated increase in the resistance of pests to
sense by keeping the different fields at an adequate the few pesticides they can use. Take, for example,
distance from each other and in an active sense by
56
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Chapter 8: “Frankenfood” 139

 crops like cotton and corn genetically modified with a thus its presence detected, under UV light.
gene from the bacterium Bacillus thuringiensis. These Finally, a concern about the complete dependence
transgenic plants then make a protein (Bt toxin) that is of agriculture on a limited number of multinationals,
toxic for insects, thus preventing them from decimating endangering the livelihood of small third-world farmers
the crops. The fear is that large-scale cultivation will in particular. Anti-cartel laws should in theory prevent
increase the risk of these insects becoming pesticide- this. And the last 30 years have shown that Third-
resistant. If, for instance, the bollworm should become World small farmers are able to adopt more efficient
resistant to the Bt toxin, it would be a disaster for technologies and have in fact done so. Nevertheless,
organic cotton growers, because bacterial Bt toxin is gene technology has made agriculture more dependent
the only means they have to eliminate this insect. In on a smaller number of large companies and we
America the mandatory solution is the so-called refuge believe it is right to ask whether this should continue.
strategy. This theory assumes that a resistant mutant
of the insect, which undoubtedly occurs once, has no 8.5. WHO IS TELLING THE WHOLE TRUTH?
advantage in relation to the non-mutated insect if the
transgenic crop is sufficiently alternated with the normal It should be obvious by now that we are not members
variant (the refuge). It sounds simple, but in practice it of the lobby that opposes gene technology. As we
isn’t. Questions such as “what percentage should be have already mentioned in Chapter 1, we have been
refuge area?”, “What is the best spatial division?”, and biotechnologists for decades. Yet, in this book, we
“What should refuge crop rotation look like?” are not are endeavouring to provide both sides with as much
easy to answer. So far the refuge strategy is working, objective information as possible. In this section we
but there is still some doubt as to whether that will will show that neither party tells the whole truth all of
continue to be the case as more of these transgenic the time, which can be very misleading for the general
crops are grown. public. For the sake of balance and not desiring to lay it
Then there is the additional concern that antibiotic- on too thickly, i.e. with a view to winning over one party
resistant genes, used particularly in the first- or another, we use just a few examples from each side.
generation transgenic crops as selection markers, There are far more detailed examples in the above-
will be transferred to bacteria, causing them to mentioned report by Rader.
become resistant to the antibiotics in question as Proponents like to say that genetic modification is as
well. There is no proof that this can actually happen, old as agriculture itself, thereby implying that there
but as a precaution alternatives are currently being is little difference between traditional breeding and
used, for example, a gene from a jelly fish, whereby breeding with the help of recombinant DNA technology.
a fluorescent protein is formed that can be seen, and It is certainly true that unexpected genetic changes

140 Part 2: Our daily food and drink

occur in the more conventional techniques, which itself have no money to buy the seed. More about Golden
should be an argument for the strict regulation of every Rice in Textbox 8.3.
newly introduced crop. It can certainly be no valid An issue about which both parties blatantly exaggerate
argument for the safety of transgenic crops to exempt in our view is food shortages. According to its
them from careful testing. proponents, biotechnology is the solution to the world’s
The big companies, who have thus far developed the hunger problems, while according to its opponents
most important transgenic crops, like to use Golden there is enough food, and the problem is simply one
Rice as their model. Golden Rice was developed with of distribution and politics. The truth lies somewhere
the aim of tackling vitamin A deficiency in large parts in-between, particularly in the future.
of the world. This shortage causes many children to And now the Pusztai affair. Árpád Pusztai (1931) is a
go blind, die prematurely or contract all kinds of other researcher born in Hungary, who has worked for most of
diseases. The vitamin A content in Golden Rice is his career at the Rowett Research Institute in Aberdeen,
dramatically increased by genetic modification. These Scotland. He is regarded as a leading expert in plant
big companies have, however, done little to develop lectins, an area in which he has published prolifically.
this transgenic rice, except for allowing their patents In 1998 Pusztai claimed in a BBC programme that the
on it to be used. This will yield little, however, since the results of his research showed that rats fed with a diet
stakeholders, i.e. the poor farmers in the Third World, of genetically modified potatoes developed deformed

TEXTBOX 8.3. Genetically modified rice, known as Golden Rice

Golden Rice. because of its colour, was developed with the aim
of increasing levels of provitamin A. Golden Rice is
Every year more than two million people go blind. In regarded as the first example of a transgenic crop
60% of cases in India, China and sub-Saharan Africa that had a direct benefit for the consumer, but in fact
this is the result of a vitamin A deficiency. Education the first was the tomato Flavr Savr (see Textbox 2.2
in the area of health care, vitamin supplementation, in Chapter 2). When Golden Rice was introduced to
gardening, food and feeding programmes and the Asia, people were confronted with the same problem
use of genetically modified rice with higher levels that we might expect with transgenic sorghum in
of provitamin A (β-carotene) are possible ways of Africa (Botha & Viljoen, 2008). Namely, concerns
preventing this. The UN Standing Committee on about the environment, patents, efficacy and social
Nutrition stresses the need to integrate these measures acceptance. As far as the environment is concerned
in order to tackle food shortages .57
there is a fear that the recombinant genes will jump
over to traditional and wild rice varieties, which
57
www.unsystem.org/scn

Chapter 8: “Frankenfood” 141

 according to ecologists could have far-reaching there is some question as to whether it will be as
consequences for Asia, because rice has its origins socially unacceptable as brown rice due to cultural
there. Golden Rice is reportedly free from royalties, preferences59. Stein et al (Stein, Sachdev, & Qaim,
but various international patents on it are in the hands 2006) conclude that Golden Rice can help alleviate
of multinationals. These businesses have agreed that vitamin A deficiency, but a range of other approaches
poor farmers don’t have to pay royalties if they earn is also necessary to tackle this complex problem.
less than 10,000 American dollars from Golden Rice, They also conclude that the uptake of β-carotene
and if the rice is not exported . In practice, however, it
58
from Golden Rice with the highest content still
would be difficult for poor farmers to prove compliance needs to be scientifically verified. In short, it is still
with either condition, simply because they don’t have very questionable whether the desired objectives
the means to do so. can be achieved with Golden Rice. We refer to the
There is also some doubt as to the efficacy of Golden previously mentioned Rader website for a more
Rice in preventing vitamin A deficiency. Provitamin A detailed overview of the fierce battle that has already
(β-carotene) first needs to be converted into retinol, been waged between opponents and supporters on
the form of vitamin A that is absorbed by the body. this subject.
The result is that, at the very most, ten percent of An announcement in the 4 February 2008 issue of
the provitamin A is finally available as vitamin A. AgraFood Biotech (Anonymous, 2008b) gives us
This means that you would have to eat Golden Rice hope. According to Robert Zeigler, general director
to the equivalent of 250 g of uncooked rice per day of the International Rice Research Institute (IRRI),
in order to obtain the required quantities of vitamin farmers will probably be able to get hold of Golden
A. Conversion of β-carotene and uptake of retinol Rice by 2011. By the end of January 2008 he reported
requires the presence of lipids, especially unsaturated that the first field trials would take place that year in
fatty acids, that are not soluble in water. Brown rice the Philippines with the aim of releasing the new crop
contains both β-carotene and the required fatty in 2011, 10 years after the first developments. He
acids in the innermost layers of the husk. However, was speaking after having received a 20 million dollar
this is lost when the husk is removed to make white subsidy for the project from the Bill & Melinda Gates
rice (which is the most popular). In Golden Rice Foundation. IRRI believes this subsidy can help them
β-carotene is also present in the innermost white supply 18 million households, primarily in South Asia
part, the endosperm, but not the fatty acids required and sub-Saharan Africa, with better rice varieties,
for conversion, so the efficacy is somewhat reduced. while it is expected that the yield will rise by 50% in
In addition, since Golden Rice is yellow in colour, the next decade.

58
www.econexus.info 59
www.panap.net

142 Part 2: Our daily food and drink

organs. The activities of Greenpeace in 1996 on the specific type of protein that are present in all nuts,
quayside at Rotterdam when the first ship laden with seeds and bulbs. Some of these lectins, such as that
transgenic soya arrived had already fuelled the fire of in the red kidney bean, are poisonous to humans.
the biotechnology debate, but this revelation really set Lectins are destroyed at higher temperatures, which
the cat among the pigeons. Pusztai was fired by the is why we first need to cook beans before eating
institute shortly thereafter. We believe that this affair them. Others, such as tomato lectins, are evidently
has been blown up so much in Europe, thanks to harmless when present. Some lectins are toxic for
the convenient manipulation of the press by extreme insects and are therefore seen as a potential natural
opponents (crash courses in how to manipulate the pesticide. The lectins from snowdrops are toxic for
press are ten-a-penny on the net), that many people insects, but not for people. In his experiment, Pusztai
have developed an irrational fear of biotechnology and fed these transgenic potatoes to one group of rats and
Europe is now lagging seriously behind in this area. normal potatoes with added lectins to a control group;
Matters have been made worse by the ban on all GM both types of potato were eaten raw. Both groups of
crops which was in force for years after the Pusztai rats developed deformed organs, and there was no
affair. statistically significant difference between the two
groups; independent statisticians later confirmed this.
HOW TO MANIPULATE THE PRESS
However, Pusztai claimed that the organs of the rats
that had eaten the transgenic potatoes were more
deformed and published this research together with
Ewen in 1999 in The Lancet. He stuck to this claim
whenever the press was present. Opponents still use
this as an argument against transgenic crops, because
it was published in such a renowned journal.
In June 1999 the influential British Royal Society
published a critical analysis of Pusztai’s results,
and concluded that they were not significant for the
following reasons:

Up until 1998 Pusztai worked at the Rowett Institute, 1. Poor experimental design, possibly exacerbated
where he conducted experiments with transgenic by lack of ‘blind’ measurements resulting in
potatoes. These potatoes were genetically modified unintentionally biased results.
with the lectin gene from snowdrops. Lectins are a 2. Uncertainty about the differences in chemical

Chapter 8: “Frankenfood” 143

 composition between strains of non-GM and GM a high-standing journal. The editors of The Lancet
potatoes. argue that despite the admittedly lax methodology
3. Possible dietary differences due to non-systematic - and disregarding the serious reservations of the
dietary enrichment to meet Home Office and other referees - they had published the article with the
requirements. hope of making a constructive contribution to the
4. The small sample numbers used in experiments debate between scientists, media and the public on
testing several diets (all of which were non-standard a heavily politically-charged subject60. Miller et al.
diets for the animals used) and which resulted in regard it as a scandalous and irresponsible deceit
multiple comparisons. and a travesty of the peer review system of research
5. Application of inappropriate statistical techniques in articles. We have no choice other than to conclude
the analysis of results. that in this case the Pandora’s box was recklessly
6. Lack of consistency of findings within and between opened. We would however prefer to confine
experiments. ourselves to the conclusion by the Royal Society that
only high-quality research within legal frameworks is
However, the following was also concluded: good enough when it comes to food safety. Neither
side can argue with that, and hopefully they will work
“Although we have no evidence of harmful effects together on this.
from genetic modification, this of course does not
mean that harmful effects can be categorically ruled 8.6. IS THERE A FUTURE FOR
out. This issue can be resolved only by the necessary TRANSGENIC CROPS?
research carried out to a high standard and by full
use of the regulatory mechanisms for dealing with “Fear of biotechnology is perpetuated by activists
safety of food.” spreading propaganda that is based on zero science
… it is the misinformed informing the uninformed.”
Pusztai himself also emphasised that his findings were
preliminary and should be seen as a forerunner to Ken Hobby, President of the US Grains Council.
further research.
In the article by Miller et al. (Miller, Morandini, & We are in no doubt that genetically modified crops
Ammann, 2008) there was also a great deal of attention have a future, even in Europe, and hopefully also in the
devoted to the Pusztai affair. They wondered how it Third World. In countries such as the US, Canada and
was possible for such an inaccurate and incomplete Argentina transgenic food crops have been cultivated
study to have got past the peer review system of such
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144 Part 2: Our daily food and drink

on a large scale for years. In fact, these crops have that we are convinced. A nice example of this sort of
nothing but advantages for the farmer, and in particular development is genetically modified carrots with higher
for the multinationals who supply the seeds and levels of absorbable calcium. Morris et al. (J. Morris,
agricultural chemicals. What we need are examples Hawthorne, Hotze, Abrams, & Hirschi, 2008) describe
where there is a clear benefit for consumers and where food studies in which people and mice are fed with
it is demonstrated that the crops are as safe as all our these transgenic carrots. They show that the uptake of
other food in all aspects. If we continue to develop this calcium rises considerably compared to control groups
technology carefully, then the future will bring food fed with normal carrots. Calcium deficiency is a major
crops that give a bigger yield, are more nutritious, problem in our Western world, causing osteoporosis
healthier, tastier and better for the environment. Of especially in the elderly. This deficiency can be

TEXTBOX 8.4. enhance the taste and nutritional values of these

Chopping onions without tears. onions. The research director Colin Eady says: “What
we’re hoping is that we’ll essentially have a lot of the
Using a special technique called gene silencing, nice, sweet aromas associated with onions without that
researchers from the Crop & Food Research Institute associated bitter, pungent, tear-producing factor. This is
in New Zealand have an exciting project because
deactivated a gene in ONIONS WERE USED FOR KEEPING YOUR AIRWAYS it’s consumer orientated and
onions. As a result, these OPEN AT NIGHT WHEN HAVING A COLD everyone sees this as a
genetically modified onions can GREAT... RRRRR... good biotechnology story.”
be chopped without inducing “NO-TEAR ZZZZZ... He expects that in about ten
tears. The deactivated gene ONIONS” years time the first ‘no-tear
codes for the enzyme onion’ will appear on the
lachrymatory factor market. His colleague
synthase (LFS). When Meriel Jones of Liverpool
traditional onions are University, who designed the Suprasweet
cut this enzyme comes onion by growing it on low-sulphur soil, hails this research:
into contact with the sulphur compounds that exist in the “This is a great development. It shows how genetic
onions and these are then converted into ‘tear inducers’. engineering can lead to real benefits for both cookery
In the genetically modified onions, LFS is not made and and health. Although conventional growing has identified
thus neither are the “tear inducers”. some sweet, mild onions, this discovery will eventually
The researchers even anticipate that this will also give farmers new varieties and consumers more choice.”

Chapter 8: “Frankenfood” 145

 prevented by using genetic modification to raise the Together, we hope we are heading for an informed,
levels of absorbable calcium in fruit and vegetables safe, healthy and sustainable society.
that contain relatively little calcium. Another more
remarkable example of such developments stems 8.7. CONCLUSIONS
from New Zealand and is described in Textbox 8.4
(Anonymous, 2008b). Researchers around the world are studying how to
To date no damage to the environment has been improve crops and farming techniques to address
observed as a result of the cultivation of transgenic worldwide hunger. By breeding staple crops such as
crops. In the case of transgenic-cotton plantations in wheat, rice, maize, soya, and sorghum to be more
particular there has been a drastic reduction in the pest- and weed-resistant, more nutrient-rich and high-
use of pesticides. In short, if it is possible for opposing yielding, and more digestible, they hope to offer more
parties to tread the DNA path together, then there nutrition per hectare of farmed land. “Grain sorghum
is, in our view, a “golden (rice) future”. So it’s a pity is a very important crop in Africa”, Kent Bradford,
that high-ranking people like Ken Hobby still make director of the Seed Biotechnology Center at the
imprudent statements about the other side (see focal University of California, Davis, said in an interview
point 2 in Chapter 3). This will do little to unite the two with Clara Moskowitz of LiveScience (Moskowitz,
parties: quite the contrary. It is all the more regrettable 2008). “Unfortunately, its protein content is relatively
because this man also does and says some very indigestible – the nutrient is inefficiently metabolized.
sensible things. The quote comes from a press release There is work in trying to modify sorghum so the protein
from early 2008 in which the US Grains Council is more digestible. That would be a huge bonus.”
(USGC) launched a publicly accessible multimedia Some scientists think the real key to ending world
and interactive CD-ROM about genetically modified hunger lies in genetically modifying crops to provide
crops available on their website . In the same press
61
boons that nature cannot match. But many people
release, Hobby also says: “The lack of user-friendly question the wisdom of dabbling in complicated natural
and accessible scientific information in one place is processes that we don’t fully understand. “I think using
one of the primary reasons why modern agricultural genetically-engineered crops would not only not solve
biotechnology still provokes concerns among many the situation, but it would continue to put the food
consumers and among international legislators. supply at risk,” said Ryan Zinn, campaign coordinator
The USCG has made this CD-ROM in an attempt for the Organic Consumers Association, a non-profit
to address this.” This touched a chord with us. It is organization, in the same article in LiveScience.
precisely for these reasons that we wrote this book. “When you’re messing with the crop’s genome, you
run the risk of opening Pandora’s box.” Conversely
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146 Part 2: Our daily food and drink

Bradford told LiveScience: ‘Nobody can point to a and benefits. The risks are exceedingly small, but the
single thing to say there’s been unintended health benefits tangible.’ We fully agree with Bradford and
consequences. While it’s always possible, it’s also repeat focal point 7 of Chapter 3 stating that only
possible that breeding crops could have unintended integrated approaches can help Third-World countries,
health consequences. It’s a matter of balancing risks not just genetic engineering.

Chapter 8: “Frankenfood” 147

 8.8. SOURCES Substantial equivalence - an appropriate paradigm for
the safety assessment of genetically modified foods?
Anonymous. (2007, 23 July). McHughen lists GM regulation’s Toxicology, 181, 427-431.
“fatal flaws”. AgraFood Biotech, p. 9. McHughen, A. (2007). Fatal flaws in agbiotech regulatory
Anonymous. (2008a, 17 March). Ninety-day feeding studies policies. Nature Biotechnology, 25(7), 725-727.
satisfactory, says EFSA. AgraFood Biotech, p. 4. Miller, H. I., Morandini, P., & Ammann, K. (2008). Is
Anonymous. (2008b, 4 February). No-tear onions. AgraFood biotechnology a victim of anti-science bias in scientific
Biotech, p. 21. journals? Trends in Biotechnology, 26(3), 122-125.
Botha, G. M., & Viljoen, C. D. (2008). Can GM sorghum Morris, J., Hawthorne, K. M., Hotze, T., Abrams, S. A., &
impact Africa? Trends in Biotechnology, 26(2), 64-69. Hirschi, K. D. (2008). Nutritional impact of elevated
Demont, M., & Devos, Y. (2008). Regulating coexistence of calcium transport activity in carrots. Proceedings of the
GM and non-GM crops without jeopardizing economic National Academy of Sciences of the United States of
incentives. Trends in Biotechnology, 26(7), 353-358. America, 105(5), 1431-1435.
Domingo, J. L. (2007). Toxicity studies of genetically modified Morris, S. H. (2007). EU biotech crop regulations and
plants: A review of the published literature. Critical environmental risk: a case of the emperor’s new
Reviews in Food Science and Nutrition, 47(8), 721-733. clothes? Trends in Biotechnology, 25(1), 2-6.
Domon, E., Takagi, H., Hirose, S., Sugita, K., Kasahara, Moskowitz, C. (2008, 23 April). Radical Science Aims to Solve
S., Ebinuma, H., & Takaiwa, F. (2009). 26-Week Food Crisis. LiveScience
Oral Safety Study in Macaques for Transgenic Rice O’Riordan, T., & Cameron, J. (1994). Interpreting the
Containing Major Human T-Cell Epitope Peptides precautionary principle. Earthscan/James & James.
from Japanese Cedar Pollen Allergens. Journal of Stein, A. J., Sachdev, H. P. S., & Qaim, M. (2006). Potential
Agricultural and Food Chemistry, 57(12), 5633-5638. impact and cost-effectiveness of Golden Rice. Nature
EFSA. (2008). Safety and nutritional assessment of GM plants Biotechnology, 24(10), 1200-1201.
and derived food and feed: The role of animal feeding van Haver, E., Alink, G., Barlow, S., Cockburn, A., Flachowsky,
trials. Food and Chemical Toxicology, 46(Supplement G., Knudsen, I., Kuiper, H., Massin, D. P., Pascal, G.,
1), S2-S70. Peijnenburg, A., Phipps, R., Poting, A., Poulsen, M.,
Goodman, R. E., Vieths, S., Sampson, H. A., Hill, D., Ebisawa, Seinen, W., Spielmann, H., van Loveren, H., Wal,
M., Taylor, S. L., & van Ree, R. (2008). Allergenicity J. M., & Williams, A. (2008). Safety and nutritional
assessment of genetically modified crops - what makes assessment of GM plants and derived food and feed:
sense? Nature Biotechnology, 26(1), 73-81. The role of animal feeding trials. Food and Chemical
Kuiper, H. A., Kleter, G. A., Noteborn, H., & Kok, E. J. (2002). Toxicology, 46, S2-S70.

148 Part 2: Our daily food and drink

part three
Health has limits
 HEALTH HAS LIMITS

“Recently there has been much criticism of health care, but since 1950 infant mortality has declined by a factor of
five and the average life expectation has increased from 71 to almost 80 years.”

Hans Galjaard

Emeritus Prof. Hans Galjaard is the father of prenatal diagnostics and clinical genetics in the Netherlands. Not
only is he a proficient physician, he is also the author of the best-selling Alle mensen zijn ongelijk (All men are
different) and of the book published in 2008, Gezondheid kent geen grenzen (Health has no limits). He is also
the man behind the statement: “It’s fascinating how many new insights have been gained into the evolution of
plants, animals and humans, thanks to the genetic revolution.” The genetic revolution is also expected to cause
a fascinating turnaround in health care. Prior to the genetic revolution there were three other developments
that transformed health care. These were:
better hygiene as a result of the introduction of
sanitation systems and sewers; anaesthesia,
which enabled doctors to treat sick patients
under sedation; and finally the introduction
of vaccines and antibiotics, which prevented
and treated many infectious diseases caused
by viruses and bacteria. Antibiotics and the
genetic revolution are the focus of part III of
this book. The possibilities in health care seem
particularly boundless as regards the genetic
revolution. It is also clear that the road to a HYGIEIA PANACEA ASCLEPIUS
panacea, to a drug to cure all ills, has still not
reached its end. For the time being, therefore, The Greek God of medicine and healing Asclepius with his daughters
health does have limits! Hygieia (goddess of health) and Panacea (goddess of healing), all
three accompanied by the snake, the symbol of health.

151
9There is a tide in the affairs of men. Which, taken at the flood, leads on to fortune.
ANTIBIOTICS

This quote from the play Julius Caesar by Shakespeare sums up better than any other the history of penicillin, a substance
excreted by the mould Penicillium notatum. Penicillin was discovered in 1928 by the British scientist Alexander Fleming.
A series of compounds, belonging to one of the most important categories of antibiotics, was derived from this substance.
The enormous potential of this antibiotic, which first became obvious during the Second World War, has been realised
in all kinds of ways. Innumerable human lives have been saved by using this category of antibiotics in cases of bacterial
infection. But that’s not all. The development of the penicillin production process was a great stimulus to the progress of
modern biotechnology in general and of large-scale biotechnological processes in particular (Demain & Elander, 1999;
Demain & Sanchez, 2009; Tramper et al., 2001). Bacteria are, however, tough little rascals and can quickly build up
resistance. So there arose a sort of eternal “arms race” between bacteria and antibiotics. Modern biotechnology enables
new weapons to be developed for preventative and curative purposes, and these are also deployed against a whole
range of other diseases. A few notable developments in the field of antibiotics are the subject of this chapter.

ONE OF THE BIGGEST DISCOVERIES

OF THE 20TH CENTURY WAS ‘JAMES BOND’
BY IAN FLEMING
THAT’S PENICILLIN!
DISCOVERED BY ALEXANDER FLEMING!

J. Tramper, Y. Zhu, Modern Biotechnology – Panacea or new Pandora’s box? 153

DOI 10.3920/978-90-8686-725-7_9, © Wageningen Academic Publishers 2011
 9.1. ANTIBIOTICS: as Penicillium notatum. A few days later, before
LIFE-SAVING BIOTECHNOLOGY cleaning the dish, he took another look and discovered
that all the bacteria around the mould had disappeared
Since its discovery in 1928 many a book has been (Figure 9.1). It is to Fleming’s great credit that he
written about penicillin. This substance, which has had realised the significance of this observation, namely
a major impact on the course of history, is referred to that a substance (named penicillin by him, because
as “yellow magic” by some writers, even though it isn’t it was produced by a Penicillium mould) hindered the
actually yellow in its pure form. The story behind the growth of bacteria. It was this discovery that finally
discovery of penicillin by Sir Alexander Fleming is well earned him a Nobel Prize for Medicine in 1945.
worth repeating. Fleming was a scientist studying the However, he made little contribution to developing it
growth of bacteria. One day one of his bacteria cultures, into a medicine. This was done by Howard Florey and
a Staphylococcus, which he was growing in Petri Ernst Chain who shared the Nobel Prize with Fleming
dishes, was contaminated by a mould, later identified (Landsberg, 1949).

Figure 9.1. Petri dish with fungus (white circles) and bacteria cultures (smears). (Source: Fotolia).

154 Part 3: Health has limits

It wasn’t until a decade later that Alexander Fleming’s transformed into a human bioreactor before the term
discovery that bacteria cultures disappeared in Petri even existed. Pure penicillin was obtained from the
dishes accidentally contaminated by moulds resulted urine of the fearless officers. When introducing his
in a pharmacological revolution: the synthesis of pure discovery in the United States, Florey truthfully declared
penicillin by Florey and Chain from the moulds that to the amazement of his American colleagues: “It’s a
Fleming described. The story of how Florey isolated remarkable substance, gentlemen: grown in bedpans
the first pure penicillin at the University of Oxford is and purified by the Oxonian police force” (Anonymous,
still one of the most gruesome stories in the history of 1995).
pharmacy. The moulds needed for the manufacturing Penicillin was first used on a large scale in the
seemed to thrive best in dirty hospital bedpans. At first American field hospitals in the Second World War,
it seemed impossible to get the antibiotic in pure form and led to an astonishing reduction in mortality from
from this ‘raw material’. Florey solved the problem infectious diseases. American and Canadian soldiers
by administering the thus-acquired moulds orally to introduced it to the Netherlands. Within a few years
the local police force, which was thereby collectively death from bacterial infection fell to unprecedented

THE FIRST HUMAN BIOREACTOR... 10

WAS A POLICE FORCE IN OXFORD! death from infectious diseases (per 1000 per year)

annual population growth (percentage per year)

2

AHHH, IT FEELS GREAT TO HELP!

8

1,5
6

0 60 0,5
46
40

44

48
42

56
50

54

58
52
19

19

19

19

19

19

19

19

19

19

19

Figure 9.2. Penicillin, infectious mortality and the post-war

population growth (adapted from Anonymous, 1995).

Chapter 9: Antibiotics 155

 levels in the Netherlands too (Figure 9.2). The main 9.2. THE BACTERIA FIGHT BACK
problem initially was how to meet the surge in demand.
First the moulds were grown in large quantities in ‘milk In the beginning penicillin was a first-rate remedy
churns’. Later, ever bigger bioreactors were designed. against bacterial infections and was often life-saving.
This technology facilitated the development of many However, all kinds of bacterial strains soon became
other large-scale biotechnological processes. resistant to it. This was mainly, but not exclusively,
In the Second World War the Dutch Yeast and Spirit the result of the exchange of plasmids with resistant
Factory in Delft was also working in deep secrecy genes (Textbox 2.1 in Chapter 2). Mobile DNA (so-
on the development of a production process for called jumping genes), bacterial viruses and the
penicillin. Five years after the war the now Royal uptake of material from dead bacteria are also means
Dutch Yeast and Spirit Factory became one of the used by bacteria to pass on resistance to each other.
biggest penicillin producers in the world. The British In the same year that penicillin was introduced on
science historian Marlene Burns wrote a thesis an industrial scale, resistant bacterial strains were
(Burns, 2005) on this development, which reads appearing, in particular among staphylococci, which
like a pure-bred thriller. She obtained her doctorate are normal skin bacteria that can cause horrible
on the strength of it in 2005 at the University of wound infections. Eight years later penicillin was
Sheffield (The development of penicillin in the only effective against 15% of Staphylococcus aureus
Netherlands 1940-1950: the pivotal role of NV infections. Staphylococcus aureus is the notorious
Nederlandsche Gist- en Spiritusfabriek, Delft). After bacterium which is now virtually resistant to
the war the Delft factory evolved into one of the antibiotics and has led to whole hospital departments
world’s biggest penicillin producers because of a being shut down for varying periods of time; it is
mixture of fortuitous circumstances. But, according referred to as MRSA, short for methicillin-resistant
to Burns, it really boiled down to what united the Staphylococcus aureus (methicillin is a second
members of the Delft research team during the war: generation antibiotic derived from penicillin) or,
a will to succeed. So it is quite disappointing, she these days, multi-resistant Staphylococcus aureus,
writes with a suitable feel for understatement, that because this strain has become resistant to virtually
now almost sixty years later, penicillin production all antibiotics. Now not only are the staphylococci
in Delft has almost entirely stopped. In March 2005 multi-resistant, but so are, for example, pathogens
the management of DSM Gist BV (successor to the like Klebsiella, Serratia and Acinetobacter strains,
Royal Dutch Yeast and Spirits Factory) decided to causing among others infections of respiratory and
move most of its production to China and India. urinary tracts.

156 Part 3: Health has limits

 H develop resistance against practically all new antibiotics.
R N S
Only the so-called glycopeptide antibiotics - most

O
importantly among them vancomycin and teicoplanin
N
- remained free from resistance. This type of antibiotic
O
has a different mechanism of action from the penicillins.
OH Only one gene is involved in resistance in penicillins,
O
whereas there are at least five involved for resistance
Figure 9.3. Basic structure of penicillin antibiotics. against the glycopeptide antibiotics.
In the 1980s people were convinced that the
In 1959 second generation antibiotics came onto pharmaceutical industry was winning the battle against
the market, the so-called semi-synthetic antibiotics. resistance. However, this conclusion seemed a little
Penicillin consist of a nucleus and a side chain (Figure premature. In 1997 vancomycin and teicoplanin also
9.3). With the help of chemical synthesis the side chain finally failed as the antibiotics of last resort when
R of penicillin can be split off and a selection of other side resistant bacterial strains were discovered. If the spread
chains can be connected to the nucleus in its place. This of these resistant strains is not stopped, the floodgates
was the start of a whole new generation of antibiotics to will open all over the world. The pharmaceutical industry,
which bacteria were not then resistant. Two years after but particularly smaller biotechnology companies, are
their introduction there were again a few Staphylococci therefore urgently looking for alternatives, especially
aureus resistant to these new antibiotics. It seemed to since the development of a new medicine is a lengthy
be only a question of time before staphylococci would process (Textbox 9.1).

TEXTBOX 9.1. Clinical trial (5-7 years):

Phases in drug development. Phase I Healthy volunteers are administered
varying doses.
A drug undergoes approximately twelve to fifteen Phase II A small group of patients receive the drug.
years of research before it appears on the market. The therapeutic effect is compared with existing
This time period includes the following phases: medicines.
Research phase (3-5 years) Phase III As in phase II, only on a bigger scale and
New substances are made and tested on their for a longer period.
efficacy. Phase IV Once the drug has appeared on the
Preclinical trial (2-3 years) market, additional clinical research takes place
First in vitro (including tissue culture), then on animals. among patients using the drug.

Chapter 9: Antibiotics 157

 is becoming an increasingly big problem. These
THE BACTERIA FIGHT BACK
practices create an environment for microorganisms
in which resistant variants have the upper hand. This
is the case in Western hospitals. The most notorious
hospital bacterium (MRSA) has been on the rise since
the 1980s, and is especially prevalent abroad. The
situation in Dutch hospitals is not so bad because
of the strict antibiotic policy adopted here. Modern
pig sties are also a breeding ground for resistant
bacteria, because pigs are administered antibiotics in
massive doses. They were previously given as growth
In 2007 a very good book by Annet Mooij was promoters and to prevent infections, but in 2006 this
published on this matter (Mooij, 2007). It is called De practice was banned and antibiotics were no longer
onzichtbare vijand – Over de strijd tegen infectieziekten allowed to be added as standard practice to the feed.
(The invisible enemy - On the fight against infectious Nevertheless, antibiotic use in pig farming is still
diseases). According to Mooij, if there is any lesson to abundant, with the result that about 40% of pigs are
be learned from the last decade it is this: that every infected with MRSA. A quarter of pig farmers is also
advance in the fight against infectious diseases is infected. Now there are staphylococci in circulation
conditional. The major plagues from the past appeared that are resistant to all available antibiotics. If these
to be under control, but diseases like malaria, cholera infections can no longer be treated, an old and serious
and tuberculosis are now no longer on the way out problem will again rear its ugly head: wound infections.
but on the way back. In the 21 February 2008 issue An ominous prospect.”
of Nature, the article “Global trends in emerging Mooij ends her book with the following paragraph:
infectious diseases” by Jones et al. (2008) states that “In the fight against infectious diseases the human
most of the (returning) upcoming infectious diseases biological machinery is clearly missing the target.
are caused by bacteria, which is a clear reflection Whether we are able to make it in the long term
of the great numbers of recorded bacteria resistant depends on the resources we have at our disposal:
to antibiotics. Mooij also writes: “In contrast to the new medicines and vaccines, knowledge and
belief of about forty years ago, the book of infectious ingenuity. The future of humanity and microbes likely
diseases will never be closed. The widespread use will unfold as episodes of a suspense thriller that
of antibiotics, unnecessary treatments and half- could be titled ‘Our Wits Versus Their Genes’, wrote
finished treatments means that antibiotic resistance the microbiologist and Nobel Prize winner Joshua

158 Part 3: Health has limits

Lederberg. It will be a tragedy in many acts, an infinite second indicator that the clinical pipeline for antibiotics
play, but thrilling it will remain.” is virtually empty, he points out that few major
pharmaceutical companies are active in the arena of
MODERN PIG STIES ARE A BREEDING
antibacterial infectious disease. The departure and/or
GROUND FOR RESISTANT BACTERIA
partial retreat of many pharmaceutical companies from
this therapeutic area around 1990, reflects not only
a mix of economic and market projections, but also
AUG APA ADCA
MEN a partial or complete failure of research programmes
TIN
that used existing models as a basis for finding new
leads that were robust enough to become clinical
candidates. In short, according to Walsh, there are not
many new (categories of) antibiotics in the pipeline.
The article “Antibiotics: where did we go wrong?” in
Drug Discovery Today of January 2005 by Overbye
and Barrett (2005), who both work for a major
pharmaceutical company, takes an identical line. The
ANTI
BIOTICS
authors present an overview of antibacterial medicines
that are in the clinical trial phase (Textbox 9.1), e.g.
9.3. THE PROSPECTS for major pharmaceutical companies three in Phase I,
one in Phase II and four in Phase III, and for smaller
A gloomy outlook, then, as far as infectious bacterial biotech companies two in the preclinical phase, four
diseases are concerned. Let us therefore summarise in Phase I, three in Phase II, one in Phase II-III, and
the pertinent trendsetting review articles from the first two in Phase III. They conclude that there has been
decade in this new millennium and see if these present a shift in antibacterial R&D efforts from the major
a more positive picture. pharmaceutical companies to a whole contingent of
Christopher Walsh put the question in Nature Reviews biotech companies. This small business approach
Microbiology of October 2003 (Walsh, 2003): “Where has led to an explosion in creativity as regards
will new antibiotics come from?” He maintains that strategies, choice of targets, use of genomics, and
there was an innovation gap of almost 40 years after development paradigms. The trends they see include
1962, before the arrival of two new categories of (1) a combination of acquisition of niche products that
antibiotics with different antibacterial mechanisms. are not being developed by major pharmaceutical
But he doubts whether this has closed the gap. As a companies, (2) the use of scientific discoveries that

Chapter 9: Antibiotics 159

 have not been applied successfully in the search scientific results, but as yet no new potential agents to
for medicines by major pharmaceutical companies, fight bacterial infections. None of the biotech companies
and (3) an increasing improvement in an existing who got onto the band wagon has survived on the
category of antibiotics. To their surprise none of the basis of its own internal R&D programme. Those who
major pharmaceutical companies has successfully did survive have various common characteristics, e.g.
developed these new target approaches in order to a solid and well-defined work plan, excellent scientific
identify potential medicines. leadership, a unique platform for a known product,
In their analysis of where ‘they’ went wrong, there are heavily financed starting capital and little income. They
about eight factors, but they stress that finger-pointing also have or had the virtually unattainable objective
is not the solution. The rapidly rising resistance of bringing an internal, potential antibiotic onto the
requires the industry to join forces and undergo far- market. The big problem with this is the enormous
reaching paradigm shifts in relation to how antibiotics amount of capital needed for the clinical trials.
can be developed and brought onto the market. In As we’ve already seen, the biotech companies have a
their view a possible way forward is to admit that “we” dozen antibiotics sitting in the pipeline, and according
have fallen short of the target (by a process of gleaning to Barrett these are to be launched over the next six to
the lessons learned from each other from previous seven years. A notable example is ceftobiprole, which
experiences) and that we must continue to look for a is currently the leading compound in a new generation
joint, universal solution to convince the pharmaceutical of cephalosporins (Figure 9.6). In March 2008 this
industry to invest again in antibacterial research and compound received the approval letter from the
development. As to the question “Where did we go FDA for the treatment of complicated skin infections
wrong?”, there is no one answer, but there must be (Cornaglia & Rossolini, 2009). Barrett expects that
one common solution. really new genomics-based developments will not be
Six months later, in 2005, Barrett (Barrett, 2005) alone implemented until around 2015. However, the average
posed the following question in Current Opinion in failure rate of medicines in this stage of development
Microbiology: “Can biotech deliver new antibiotics?” means that no more than two to four of these candidate
He reported that after the publication in 1995 of the first antibiotics will reach the market, and none of them are
two complete genome sequences of two pathogenic expected to be a great success. But in Barrett’s view
bacteria (Haemophilus influenzae and Mycoplasma it does show that the biotech companies could deliver
genitalium) many academic and industrial laboratories such products, in isolation from the R&D paradigm, and
threw themselves into pathogenic bacterial genomics it is this potential success that will propel the continued
with the aim of developing new anti-bacterial methods investment. Basically, modern biotechnology can
and/or antibiotics. This has delivered many spectacular deliver antibiotics, but it must be done in collaboration

160 Part 3: Health has limits

with the major pharmaceutical companies so that the here is that it was written by the medical specialists
costly clinical trials can be funded. Salvador Bello Drond and Manuel Vilá Justribó, who
In October 2003 Walsh asked where the new antibiotics work in the Spanish university hospitals of Zaragoza
would come from (Walsh, 2003). In December 2006 and Lleida, respectively. Even more interesting is that
he and two colleagues (Clardy, Fischbach, & Walsh, Spain is one of the countries that is a real breeding
2006) answered this question in Nature Biotechnology: ground for resistant bacteria because of the excessive
“New antibiotics from bacterial natural products.” The and practically unlimited use of antibiotics.
general gist of their article is as follows. The demand These two medics are well aware of this and wrote the
for new antibiotics has largely been met in the last article as a warning. They claim that because of the
five decades by semi-synthetic customisation of fast growth in the number of multi-resistant bacteria,
a few natural molecular template structures, such fears are growing among doctors and that this fear is
as penicillin, which were discovered in the mid- gradually starting to filter through to society as a whole.
20th century. More recently, however, technological In their view, the problem is made worse by the fact
advances, e.g. the introduction of high-throughput that very few people are aware that there is little hope
screening techniques, have seen a reincarnation of the at this moment of many new antibiotics coming on the
search for antibiotics from bacterial natural products. market in the short to medium term. The authors advise
The main focus of the search is for new antibiotics that careful use of the available antibiotics, based on
from old (e.g. streptomycetes) and new sources (e.g. a detailed knowledge of their mechanism and the use
actinomycetes, cyanobacteria and as yet uncultivated of new forms of administration, such as inhalers, may
bacteria). This has resulted in various newly solve the problem in part. A very sound piece of advice,
discovered antibiotics with unique template structures particularly, but certainly not exclusively, for a country
and/or new mechanisms, with the potential to form the like Spain. The article “How antibiotics can make us
basis of new categories of antibiotics. In their list of sick: the less obvious adverse effects of antimicrobial
these antibiotics which are already in the clinical trial chemotherapy” from 2004 by Stephanie Dancer in
phase, we see practically the same substances that The Lancet Infectious Diseases also carried the same
we saw in Barrett’s table. In many cases, though, a message (Dancer, 2004).
major pharmaceutical company has come on board. Another notable review article on antibiotics we found
An example of this trend is described in Textbox 9.2. was called “Combination drugs, an emerging option
for antibacterial therapy.” It was written by Cottarel and
Yet another interesting review article, called “Will We Wierzbowski (Cottarel & Wierzbowski, 2007) from the
Still Have Antibiotics Tomorrow?”, was published in Center for Advanced Biotechnology, Boston University,
2007 (Dronda & Justribo, 2007). What’s interesting and was published in December 2007. The authors

Chapter 9: Antibiotics 161

 TEXTBOX 9.2. and fungal cell walls. At that time the company worked
Vicissitudes of a typical anti-infectives biotech together with Wageningen University, the Netherlands,
company. on the cleavage of the side chain of A40926 (Figure 9.4).
The product can be used as template to prepare new
Lepetit Research Centre was until 1995 a medium-sized glycopeptide derivatives (Jovetic, de Bresser, Tramper,
(100-150 employees) research laboratory, belonging & Marinelli, 2003). In 2000 Biosearch Italia became
to Marion Merrell Dow, devoted to the discovery and the first small biotech company in Italy to go public and
development of novel anti-infectives. It was established appear on the Nuovo Mercato stock exchange. Then in
in Gerenzano near Milan, Italy. Lepetit discovered 2003 it merged with the American biopharmaceutical
rifamycin followed by teicoplanin, ramoplanin, lantibiotic company Versicor Inc. into Vicuron Pharmaceuticals
actagardine, A40926 and dalbavancin, all important (listed on both the NASDAQ and Nuovo Mercato stock
antibiotics. In 1995 Lepetit was bought by Hoechst exchange). At that time the company had three molecules
and became Hoechst Marion Roussel, representing in the clinical pipeline (Phase II and III), i.e. dalbavancin,
the pharmaceutical branch of Hoechst. As result of anidulafungin and ramoplanin. In 2005 the company was
this operation Biosearch Italia arose as a spin-off in bought for a stunning $ 1.9 billion by Pfizer, who brought
1997. Biosearch Italia presented itself as a small Italian anidulafungin onto the market as a novel antifungal for
biopharmaceutical company focusing on new antibiotics systemic infections. By late 2006 Pfizer implemented
for the treatment of infections caused by multi-resistant a global R&D restructuring and closed the research
pathogens. They focused on glycopeptides, the class of centre in Gerenzano. However, molecules discovered
antibiotics to which vancomycin, teicoplanin, A40926 and by Lepetit scientist continue their story. Ramoplanin,
dalbavancin belong, and on other inhibitors of bacterial since December 2009 acquired by Nanotherapeutics,

A. teichomyceticus

A40926 deacyl-A40926

Figure 9.4. Bioconversion of A40926 into deacyl-A40926.

162 Part 3: Health has limits

 Inc., based in Florida, is now entering Phase III trials as glycopeptide, has been recently acquired by a newly
an oral antibiotic for the treatment of Clostridium difficile- formed US-based biopharmaceutical company Durata
associated disease (Shah et al., 2010). Dalbavancin Therapeutics, Inc., and is proceeding in its late stage
(Malabarba & Goldstein, 2005), a second generation clinical development.

conclude like everyone else that there is an urgent infection. Another advantage of this approach is
need for new effective therapies for the treatment of that it may result in shorter treatment periods and/or
infectious bacterial diseases, whereby the increase in lower doses, which may reduce the speed with which
resistance is minimised. In their view, combinations pathogenic bacteria become resistant. There are now
of different categories of antibiotics or the addition a few small (start-up) biotech companies working on
of adjuvants (pharmacological agents that reinforce the development and marketing of such antibacterial
the antibacterial action) are a promising alternative combinations.
therapeutic approach whose efficacy has already The authors divide the combination therapies into four
been proven in, for example, tuberculosis. Starting categories on the basis of the mechanism of action
with the existing categories of antibiotics (Table 9.1), with which the components potentiate the activity (of
it is not only possible to increase the activity of well- each other) (Figure 9.5):
known and effective antibiotics with combinations, but A

also to support the development of substances which

A 2a
have already proved to be very effective antibiotics A

but which are too toxic for patients with a bacterial 2b

T1 4a
1

Table 9.1. Most important classes of antibiotics and their action. T2 4b

Class Action 3
Fluoroquinolones Inhibition of DNA synthesis
A
Aminoglycosides, tetracyclines, Inhibition of protein synthesis

ketolides, macrolides, chloramphenicol,

lincosamides Figure 9.5. Mechanisms of combination therapy: (1) Adjuvant (A)

Rifampicin Inhibition of RNA synthesis inhibits the degradation or modification of the drug; (2) adjuvant
Trimethoprim, sulfonamide Inhibition of folic acid synthesis inhibits the cell repair (a) or intrinsic resistance pathway (b);
Penicillins, cephalosporins, Inhibition of cell wall synthesis (3) adjuvant inhibits the efflux pumps; (4) combination of two
carbapenems, daptomycin antibiotics with (a) or without (b) similar target T, reproduced
Colistin, polymyxin Damage to cell wall integrity with permission (Cottarel & Wierzbowski, 2007).

Chapter 9: Antibiotics 163

 1. The breakdown or modification of the actual 9.4. ‘GREEN’ PRODUCTION
antibiotic is prevented by a second compound, the
adjuvant. Although it seems from the above that semi-synthetic
2. The accumulation or retention of the actual antibiotic antibiotics have a limited life, they are still produced and
in the cell is facilitated by an adjuvant which stops it prescribed on a large scale. The “big” antibiotics such
being pumped out. as ampicillin, amoxicillin and cefalexin are expected
3. The intrinsic repair and tolerance mechanism of the to be in use for a further 20 years. The first two are
bacterial cells against the antibiotic is inhibited by penicillins, the third belongs to the cephalosporins,
an adjuvant. which are derivatives of penicillin (Figure 9.6). DSM-
4. A second compound is itself also an antibiotic with Gist is the global market leader in this area. The
the same or a different mechanism of action from biggest competition comes from Spain, Italy, India and
the primary antibiotic. China. The processes used in these countries start
with the fermentation product penicillin G obtained
Some of these combinations have already been from traditional moulds. This is used to make the semi-
used successfully to fight difficult infections. The manufactures 6-APA and 7-ADCA from which various
most well-known example is Augmentin which was
®
semi-synthetic antibiotics are then manufactured,
brought onto the market by GlaxoSmithKline. The including the above-mentioned ones.
antibiotic in this is amoxicillin belonging to the β-lactam In the Dutch DSM production process of cefalexin, which
(penicillins) category of antibiotics which inhibit cell began in March 2001 in Delft (the former Gist-brocades),
wall synthesis. Bacteria can become resistant to this a genetically modified Penicillium strain is used, which
by taking a gene from other organisms that codes dramatically reduces the number of processing steps
for an enzyme that breaks down β-lactam. These and thus the costs. The new process also uses 35%
enzymes, the β-lactamases, catalyse the hydrolysis less energy and the use of organic solvents has been
of penicillins causing them to become ineffective. If a reduced to virtually zero. By using genetically modified
pathogenic bacterium takes up a gene which codes for moulds and by replacing chemical synthesis with
a β-lactamase that can break down amoxicillin, it has biocatalysis (i.e. the use of enzymes as biocatalysts) in
then become resistant to it. Clavulanate, a compound the subsequent steps too, the production of antibiotics
that inhibits these β-lactamases, is present beside has become much more environmentally friendly;
amoxicillin in Augmentin®. As a result amoxicillin can still hence the name ‘green’ chemistry. As mentioned in
have an antibacterial effect on these resistant bacteria. Section 9.1, all antibiotic production processes at DSM
All in all, not a very rosy outlook, but luckily this have been transferred to India and China. Only the
Pandora’s box is not completely without hope! production process for cefalexin with the genetically

164 Part 3: Health has limits

 modified mould is still carried out in the factory in Delft. consists of more than 100 scientific publications and
DSM is not only the biggest producer of these various patents; Figure 9.6 comes from this book. DSM
antibiotics, it is also leading the way in terms of also conducts a lot of molecular biological and genetic
knowledge (generation). A large-scale study in the research on moulds and in 2005 they unravelled the
form of an intensive collaboration between DSM and gene card, the genome, of Penicillium chrysogenum.
six Dutch academic research groups over a five-year This is not the strain that Fleming used for his discovery.
period (the Chemferm project) ended in 2001 with the It is the modern workhorse for penicillin production, and
publication of a book edited by Alle Bruggink. The book has since been ‘bred’ so that it produces a thousand
contains a summary of the results of this teamwork and times more penicillin than its natural predecessor which

Fermentation was plucked from a melon in 1953. Until recently DSM

were the exclusive owners of the genome sequence,
H but now, 80 years after the discovery of penicillin, they
N S
Me have published it (van den Berg et al., 2008), probably in
O N Me
O order to pass the post before other researchers. Either
COOH
Penicillin G way, it opens the way to yet more innovative processes
H2 N S H2N S and antibiotics based on modern biotechnology.
Me
N Me N
O O Me
COOH
NH2 COOH 9.5. A NEVER-ENDING STORY
6-APA R’ 7-ADCA
O
R It seems highly likely that the battle between bacteria

NH2 H NH2 H
and antibiotics will become a never-ending story. This
N S
Me
N S seems to hold true today more than ever. New medicines
O Me O
R O
N R O
N Me will continue to be followed by new resistances. Global
COOH COOH
public health then becomes a matter of which is faster,
SSP’s SSC’s
the bacteria or the pharmaceutical industry. Given the
Figure 9.6. General production chart of penicillin-derived time required to build up resistance and to develop a
antibiotics (semi-synthetic penicillins, SSP’s, e.g. ampicillin medicine, this looks set to be and is in fact already an
(R = H) and amoxicillin (R = OH)) and cephalosporin-derived exciting race. As we know, a few biotech companies
antibiotics (SSC’s, e.g. cefalexin (R = H) and cefadroxil (R = have already started on new experimental methods
OH)). Intermediates: 6-aminopenicillanic acid (6-APA) and to tackle resistant bacteria once and for all. Recently
7-aminodesacetoxy-cephalosporanic acid (7-ADCA). Reproduced we have published a review on this topic (Jovetic et
with permission from Tramper et al. (2001). al., 2010). Many pharmaceutical companies have also

Chapter 9: Antibiotics 165

 taken the plunge, literally and figuratively, looking for by binding to a specific location on a bacterium protein
new types of antibiotic and other biological activities. which is essential for the survival of this bacterium.
According to Williams (Williams, 2009) marine The researchers use the interaction between these
bacteria will equip us in the coming century, if properly two as a basis for a method to quickly trace new
developed and used, with weapons for our eternal antibiotics. The researchers speculated that a
battle against multi-resistant bacteria. chemical substance that can interrupt the interaction
The sea is our richest source of biodiversity and is, between an antibacterial phage protein and an
so far, practically unexploited. Sponges in particular essential bacterium protein may well have the same
demonstrate a wealth of biological activity that is antibacterial effect. After a screening of 125,000 small
promising for medical use. ‘Overfishing’, however, molecules, a targeted selection of a phage protein
threatens these very vulnerable ecosystems. It is and Staphylococcus aureus protein provided eleven
therefore vital to develop technologies to prevent this candidate antibiotics; these indeed seemed to be
occurring. That is why a number of researchers from able to inhibit the growth of Staphylococcus aureus.
our own research group are working, for example, Small molecules are better antibiotics than the
on bioreactors, in which they plan to cultivate huge relatively big intact bacteriophages and the relatively
quantities of sponges from very small quantities big antibacterial phage protein molecules. They have
under tightly controlled conditions; this being the better pharmacokinetic properties in human tissues
first criterion for developing a similar pharmaceutical and are less likely to cause undesirable immune
process (Sipkema et al., 2005). It is sometimes also reactions. Bacteriophages and antimicrobial phage
worth first taking a look back in history and revisiting proteins seem to be valuable instructors in any case
old knowledge in the light of what we know now. This is in the search for this sort of ‘small’ antibiotic. Yet
dealt with in the following section of this chapter. the use of bacterial viruses as a cure for bacterial
infections dates back to 1921 and was the discovery
9.6. TAKING ANOTHER LOOK AT PHAGES of the Frenchman Félix d’Herelle. This therapy was
however consigned to oblivion, except in the former
Bacteriophages or “bacterial viruses”, usually called Eastern bloc, because of the meteoric rise of the
phages, are natural specialists in killing bacteria. To antibiotics. There researchers worked eagerly on
do this they produce a whole range of antibacterial the further development of phage-based medicines
proteins. These phage proteins may be a source of (Vandamme & Raemaeckers, 2003).
inspiration in the search for new antibiotics, according Independently of each other, Edward Twort in 1915
to a study by Canadian scientists (Projan, 2004) A and Félix d’Herelle in 1917 discovered the phages and
phage protein exercises its antibacterial mechanism d’Herelle was the first to use the term bacteriophage.

166 Part 3: Health has limits

Like all other viruses, phages consist of an outer from the rapidly increasing antibiotic resistance. A
shell of proteins enveloping a DNA or RNA strand, Western ode to bacteriophages appeared in the 2007
their genome. Viruses cannot translate or copy their article “Biotechnological exploitation of bacteriophage
genetic material themselves. They use the “expertise” research” (Petty, Evans, Fineran, & Salmond, 2007).
and “machinery” of other organisms for this purpose. Aside from the huge possibilities for molecular biology,
Viruses reproduce at the expense of cells of living nanotechnology and the detection of specific bacteria,
organisms. In the case of phages these are bacteria. the authors observed a whole shopping list of potential
They are not interested in animal and plant cells. In opportunities for the use of phages to treat infectious
theory, therefore, bacteriophages seem to be the ideal bacterial diseases. In short, the phage therapies are
agent for treating infectious bacterial diseases. no longer on the way out, but are on the way back, as
That’s what they thought in the former Soviet Union, Annet Mooij concluded in Section 9.2 for the “enemy”,
where research into phage medicine was carried out better known as infectious bacteria. One thing is
in earnest. In the period 1920-1950 no fewer than 800 certain: phage-based antibacterial medicines still have
mostly Russian publications on phages appeared. a long way to go in the research centres before we’ll
The Eliava Institute in the Georgian capital Tbilisi, find them in (Western) pharmacies.
which focused entirely on phage therapy, had its
heyday between 1970 and 1980. Hundreds of people 9.7. CONCLUSIONS
worked on the production of phage medicines and
huge quantities of phages in the form of pills, creams Medical care requires constantly novel antibiotics due to
and sprays were sold over the counter. The collapse the growing prevalence of resistant pathogens in hospital
of the Soviet Union and the ensuing economic crisis or community-acquired infections. Notwithstanding
also signalled the downfall of the Eliava Institute. It is this need, major pharmaceutical players seem to be
now trying to survive on the back of a few spin-offs reducing their R&D efforts in the area of new antibiotics.
and some phage preparations are still being produced, This is due to a combination of factors: considerations
for instance an artificial skin which is impregnated with about maturity of the new drug candidates, the strong
viruses to heal skin and burn wounds. competition among pharmaceutical companies, and the
The West was never convinced by the Georgian phage increase in generic antibiotics on the market. There is
therapy and still isn’t. From a scientific perspective, still a perception that the discovery of novel antibiotics
this is not altogether unjustified, because there are has become a very rare event. Despite significant
still a great many snags in this area, and much more advances in bacterial genomics, high-throughput
scientific substantiation is needed. Now, however, this screening techniques and synthetic methods, the
reticence seems to be changing, partly due to pressure discovery of novel antibiotics over the past thirty years

Chapter 9: Antibiotics 167

 has not sufficiently kept pace with the demand for new resistance is an unavoidable aspect of evolution,
agents. On the other hand, past and present successes which indeed is closely linked to the magnitude of the
suggest a return to microbial products that could be selective pressure. This was once more emphasised
used per se or as scaffolds in the quest for better on the day, 11 August 2010, when we were finishing this
drugs against multiresistant bacteria. Fortunately, our chapter by the alarming news in the daily papers that a
armamentarium for treating Gram-positive infections new Asian superbug has spread from India to the UK.
is being enriched by novel β-lactams, glycopeptides, These bacteria have a newly found gene called New
lantibiotics and other peptides in different phases of Delhi metallo-beta-lactase, or NDM-1, making them
development, and our options are increasing with the highly resistant to almost all antibiotics, including the
introduction of specific vaccines and combinatorial most powerful class called carbapenems, and experts
drugs. It has never been more important to understand say there are no new drugs on the horizon to tackle it.
in detail the mechanisms of, and routes to, resistance As our battle with antibiotic resistance is thus destined
in bacteria, so that we can improve the surveillance to continue, it is of the utmost importance that we learn
of emerging mechanisms of resistance to antibiotics to use antibiotics cautiously and appropriately. Only
introduced in clinics and the environment. We should in this way can we delay the spread of resistance, a
be aware that emergences and diffusion of bacterial natural phenomenon that will surely not disappear.

168 Part 3: Health has limits

 9.8. SOURCES J., et al. (2008). Global trends in emerging infectious
diseases. Nature, 451(7181), 990-993.
Anonymous. (1995, January). Hiroshima, penicilline en de Jovetic, S., de Bresser, L., Tramper, J., & Marinelli, F. (2003).
ENIAC. PolyTechnisch Tijdschrift, pp. 36-39. Deacylation of antibiotic A40926 by immobilized
Barrett, J. F. (2005). Can biotech deliver new antibiotics? Current Actinoplanes teichomyceticus cells in an internal-loop air-
Opinion in Microbiology, 8(5), 498-503. lift bioreactor. Enzyme and Microbial Technology, 32(5),
Burns, M. (2005). Wartime Research to Post-War Production: 546-552.
Bacinol, Dutch Penicillin, 1940-1950. University of Jovetic, S., Zhu, Y., Marcone, G. L., Marinelli, F. & Tramper, J.
Sheffield, Sheffield. (2010). ß-lactam and glycopeptide antibiotics - first and
Clardy, J., Fischbach, M. A., & Walsh, C. T. (2006). New last line of defence? Trends in Biotechnology, (In Press),
antibiotics from bacterial natural products. Nature Doi: 10.1016/j.tibtech.2010.09.004.
Biotechnology, 24(12), 1541-1550. Landsberg, H. (1949). Prelude to the discovery of penicillin. Isis,
Cornaglia, G., & Rossolini, G. M. (2009). Forthcoming 40(3), 225-227.
therapeutic perspectives for infections due to multidrug- Malabarba, A., & Goldstein, B. (2005). Origin, structure, and
resistant Gram-positive pathogens. Clinical Microbiology activity in vitro and in vivo of dalbavancin. Journal of
and Infection, 15(3), 218-223. Antimicrobial Chemotherapy, 55(Supplement 2), ii15-ii20.
Cottarel, G., & Wierzbowski, J. (2007). Combination drugs, Mooij, A. (2007). De onzichtbare vijand - Over de strijd tegen
an emerging option for antibacterial therapy. Trends in infectieziekten. Amsterdam, Balans.
Biotechnology, 25(12), 547-555. Overbye, K. M., & Barrett, J. F. (2005). Antibiotics: where did we
Dancer, S. J. (2004). How antibiotics can make us sick: the less go wrong. Drug Discovery Today, 10(1), 45-52.
obvious adverse effects of antimicrobial chemotherapy. Petty, N. K., Evans, T. J., Fineran, P. C., & Salmond, G. P. C.
Lancet Infectious Diseases, 4(10), 611-619. (2007). Biotechnological exploitation of bacteriophage
Demain, A. L., & Elander, R. P. (1999). The beta-lactam research. Trends in Biotechnology, 25(1), 7-15.
antibiotics: past, present, and future. Antonie Van Projan, S. (2004). Phage-inspired antibiotics? Nature
Leeuwenhoek International Journal of General and Biotechnology, 22(2), 167-168.
Molecular Microbiology, 75(1-2), 5-19. Shah, D., Dang, M., Hasbun, R., Koo, H., Jiang, Z., DuPont,
Demain, A. L., & Sanchez, S. (2009). Microbial drug discovery: H., et al. (2010). Clostridium difficile infection: update
80 years of progress. Journal of Antibiotics, 62(1), 5-16. on emerging antibiotic treatment options and antibiotic
Dronda, S. B., & Justribo, M. V. (2007). Will we still have resistance. Expert Review of Anti-Infective Therapy, 8(5),
antibiotics tomorrow? Archivos De Bronconeumologia, 555-564.
43(8), 450-459. Sipkema, D., Osinga, R., Schatton, W., Mendola, D., Tramper,
Jones, K., Patel, N., Levy, M., Storeygard, A., Balk, D., Gittleman, J., & Wijffels, R. H. (2005). Large-scale production

Chapter 9: Antibiotics 169

 of pharmaceuticals by marine sponges: Sea, cell, or J., Driessen, A., et al. (2008). Genome sequencing
synthesis? Biotechnology and Bioengineering, 90(2), and analysis of the filamentous fungus Penicillium
201-222. chrysogenum. Nature Biotechnology, 26(10), 1161-1168.
Tramper, J., Beeftink, H. H., Janssen, A. E. M., Ooijkaas, L. P., Vandamme, E., & Raemaeckers, P. (2003). Virus komt de mens
Van Roon, J. L., Strubel, M., et al. (2001). Biocatalytic te hulp. Natuur, wetenschap en Techniek, 26(September),
production of semi-synthetic cephalosporins: process 29.
technology and integration. In A. Bruggink (Ed.), Walsh, C. (2003). Where will new antibiotics come from? Nature
Synthesis of β-lactam antibiotics - Chemsitry, Biocatalysis Reviews Microbiology, 1(1), 65-70.
and Process Integration. (pp. 207). Dordrecht, Kluwer Williams, P. G. (2009). Panning for chemical gold: marine
Academic Pubishers. bacteria as a source of new therapeutics. Trends in
van den Berg, M., Albang, R., Albermann, K., Badger, J., Daran, Biotechnology, 27(1), 45-52.

170 Part 3: Health has limits

10 HORMONES: NATURAL REGULATORS

“We are not doctors and we aren’t writing prescriptions for you! We believe that we are smarter than most doctors
about steroids. We’re sticklers about the truth in anything and we happen to know a lot about steroids (some say that
we know too much). This book is telling you what we believe to be practical, real world information incorporating the
very latest developments in steroid use. You may not care for our sense of humour, or our attitudes, but we honestly
think that there is very little argument in the factual information presented. We happen to be bodybuilders so we do
slant the information toward that endeavour. What’s important is that most of the drugs we talk about, we’ve used
ourselves a number of times. You should know how a drug really works, not how the label says it’s supposed to.”

An excerpt from the original Underground Steroid Handbook62; Daniel Duchaine wrote the book in 1988, just before
he went to prison for a year on a steroid charge.

In order for a body to function well, be it that of a human or animal, the many chemical reactions that take place
in the cells need to be well regulated and in tune with each other, as do those of the various tissues and organs.
Hormones have an important role to play here. They regulate metabolism, reproduction, growth and many other
bodily processes. Behaviour and frame of mind are also affected by them.

HORMONE USE CAN HAVE STRANGE EFFECTS

WE BELIEVE WE’RE SMARTER

THAN DOCTORS ABOUT STEROIDS ...
... I EVEN WROTE A
BOOK ABOUT IT!

MY FIR
STEROIDST
S

62
www.qfac.com/books/origush.html

J. Tramper, Y. Zhu, Modern Biotechnology – Panacea or new Pandora’s box? 171

DOI 10.3920/978-90-8686-725-7_10, © Wageningen Academic Publishers 2011
 10.1. WHAT ARE HORMONES? waste products, such as hydrogen peroxide, are broken down;
(11) cytoplasm, living content of a cell, not including the nucleus
Hormones come in all shapes and sizes. Some are and large vacuoles; (12) lysosome, an organelle that contains
steroids, complex chemical compounds, such as a number of enzymes, whose destructive capacity means
our sex hormones (progesterone, testosterone and that they have to be separated from the rest of the cell; (13)
oestrogen), but bile acids and sterols (e.g. cholesterol) centrioles, organelles that play an important role during nucleus
division; (14) membrane, selectively permeable structure,
14 9 13 10 8 7 12 3 1 2 5 6 4 11 composed mainly of lipids and proteins, which surround cells
and also occur within the cells to encase organelles.

also belong to this group. Others are shorter or longer

chains of amino acids, peptides and proteins, such
as insulin. There are differences between hormones
from glands, tissues and cells. The first are produced
in the glands and transported via the bloodstream
to the organs where they do their work. The tissue
hormones only exercise their influence in their close
Figure10.1. Cross-section of an animal cell: (1) nucleolus, a surroundings. Cell hormones regulate all processes
non-membrane bound structure that produces ribosomal RNA; in the cell which they inhabit. Hormones are also
(2) nucleus, cell nucleus containing the DNA; (3) ribosomes, categorised according to their mechanism of action
small organelles where protein synthesis occurs; (4) vesicle, a or chemical structure. They are identified by specific
small vacuole, for example, a Golgi vesicle or a membranous molecules or receptors in the cells of their target
vesicle, for transporting larger quantities of material; (5) organ. The receptors are usually proteins at the cell
rough endoplasmatic reticulum, for transporting proteins, with membrane (e.g. receptors for insulin or adrenalin)
ribosomes on the surface; (6) Golgi apparatus, network in which or in the cytoplasm (e.g. receptors for oestrogen or
products like polysaccharides are produced and taken away progesterone). The hormones regulate by latching
by budded vesicles; (7) microtubule, cylindrical unbranched onto enzymes or nucleic acids (the building blocks of
tube that fulfils a skeletal function in cells that are not round, the genetic material, DNA and RNA). Plants also have
for example, nerve cells; (8) smooth endoplasmatic reticulum, such regulators, called phytohormones.
carries no ribosomes and is involved in fat metabolism; (9) When a hormone is not produced in the correct
mitochondria, function in aerobic respiration and generate quantities in an organism, various anomalies occur.
energy for the cell; (10) peroxisome, microbody where toxic Sometimes the changes are a natural process; for

172 Part 3: Health has limits

instance, a reduction in oestrogen levels during mass. Growth hormone deficiency in adults can also
the menopause in aging women or a gradual have a great many serious consequences.
decline in testosterone levels in the andropause Following the discovery and development of growth
or male menopause in aging men. Where there is hormone into medicines, pituitary glands from
a deficiency, it is possible in some cases to top-up deceased people were for many years the only source
hormones using medicines. The first example of this from which this hormone could be isolated. The use of
was human insulin made from recombinant bacteria, the thus acquired somatotropin was strictly regulated
already described in Chapter 1. In the present chapter and only prescribed in children with a serious form
three other examples of human hormones are of dwarfism as a result of a lack of self-made growth
discussed, namely growth hormones (somatotropin hormone. In general the children who were treated
or somatrophin), erythropoietin (EPO) and follicle- underwent a surprising recovery in growth and were
stimulating hormone (FSH). All three are products of consequently spared the (psychological) misery
modern biotechnology. of dwarfism. As a rule-of-thumb the treatment was
started before puberty and stopped as soon as the
10.2. HUMAN GROWTH HORMONE (HGH) epiphyses, the cartilaginous endplates of the bones,
had fused (Pownall, 1994).
Growth hormone as medicine Tragically, however, it became clear that the
Human growth hormone has been firmly placed on the therapeutic use of growth hormone acquired in this
medical map since the end of the 1950s. At that time way also brought with it major risks, e.g. the transfer
clinical researchers discovered this hormone in the of diseases. The most horrific of these is Creutzfeldt-
pituitary, a gland the size of a small pea in the middle Jakob disease (caused by prions and comparable to
of the head under the brain. The pituitary is regarded Mad Cow Disease), which raised serious questions at
as the ‘master gland’ because many hormones from the beginning of the 1980s about the use of growth
there regulate the excretion of hormones in other hormone extracted from pituitaries. In 1985 the use
glands. In times gone by, the pituitary was called of this hormone in the Netherlands was discontinued.
the seat of the soul. Hormone deficiency can be In an article entitled “Illegale hormonen” (illegal
congenital or the result of a cyst, tumour, radiation hormones) which was published as a scientific
or trauma. The consequences for children and adults editorial in the NRC on 18 July 2002, the following
can be far-reaching. For every ten thousand births, appeared. To their knowledge, one of the more than
one newborn baby suffers from a growth hormone 560 children treated with growth hormone extracted
deficiency of some degree of severity. These children from the pituitary gland of deceased people has died
grow (much) slower and have (a lot) less muscle of Creutzfeldt-Jakob disease. Fortunately, the sale of

Chapter 10: Hormones: natural regulators 173

 human growth hormone, obtained from recombinant
E. coli bacteria, has now made the bizarre extraction
from human pituitary glands unnecessary. This method
was introduced in 1985, shortly after insulin came onto
the market as the first hormone product of recombinant
DNA technology. The commercial availability of much
bigger quantities of safe growth hormone has also
opened the way to other applications, not all of which
have been equally desirable, as we will see in the
following paragraphs.
New research has meantime shown that the fusion
of the epiphyses does not signal an end to the need PRESENT MARKET: FUTURE MARKET:
for growth hormone. Adults with growth hormone € 1.5 BILLION € 3 BILLION?
deficiency often have all kinds of deficiencies, such Figure 10.2. Expected trend in demand for human growth
as an abnormal physical make-up, a poorer physical hormone.
condition, a different fat metabolism (accumulation
of fat tissue), increased cholesterol levels and as a Growth hormone as anti-ageing agent
result more cardiovascular disorders, porous bones, Children and adults with growth hormone deficiency
sexual disorders, and an overall reduced quality of life have relatively large amounts of fat and very little ‘lean’
and reduced life expectancy, even if they have been tissue mass. Their muscles are poorly developed and
treated with growth hormones as children. Clinical they have little stamina, while their kidney function is
studies carried out in the late 1990s convinced the impaired and their blood pressure is low. This is proof
American authorities to approve growth hormone that growth hormone does more than just regulate
treatment of adults with a deficiency. According to Ken the growth of a child. When adults with a deficiency
Attie, a clinical researcher who worked at Genentech receive growth hormone, there are recorded reductions
(a manufacturer of recombinant growth hormone) in in subcutaneous and abdominal fat of 13 and 30%
San Francisco at the time of his pronouncement, in respectively. More muscle tissue is also developed.
the US alone there are 70,000 adults with a growth The media has exaggerated these types of results
hormone deficiency (Pramik, 1999). This alone, and made growth hormone out to be a sort of elixir of
however, is insufficient to explain the growth in youth. However, non-prescribed and unsupervised use
demand (Figure 10.2), as predicted at the end of the is very unwise, because of the potential occurrence of
20th century. all manner of side effects such as fluid accumulation

174 Part 3: Health has limits

(oedema), headache and an unhealthy reduction in ageing?” (Vance, 2003). She reported that there were
blood sugar levels (hypoglycaemia), especially at high various websites offering growth hormone in oral or
doses. aerosol form. In her opinion efficacy has not been
In 1990 an article was published on the effects of shown in any of these substances. She concludes
human growth hormone on men over 60 (Rudman that follow-up studies confirm the findings of Rudman
et al., 1990). It suggested that a short course of et al. concerning changes in physical condition, but
recombinant growth hormone could reverse ageing improvements in functioning have not been observed.
symptoms, such as paunch, atrophied muscles, In her view fitness has a more positive effect.
double chin and reduced sexual performance, in
GROWTH HORMONE IS USED FOR ANTI-AGEING
otherwise healthy men. The article was based on a six-
month study of twelve elderly men ranging from 61 to I’M 90 YEARS OLD, BUT STILL
81 years of age. As a result of this article rejuvenation
LOOKING FANTASTIC
anti-ageing clinics offering the human growth hormone
(HGH) sprouted up all over the place, particularly in
the US, and popular science books and articles were
published with enticing titles like:

“Grow young with HGH: the amazing medically proven

plan to reverse aging” (Klatz & Kahn, 1998).
“HGH: Age-reversing miracle” (Elkins, 1999).
“Staying young: growth hormone and other natural In 2007 a review article on this appeared in the
strategies to reverse the aging process” (Gilbert, renowned journal published by the American College
Jamie, & Gross, 1999) of Physicians (Liu et al., 2007). The seven authors, all
“The new anti-aging revolution: stopping the clock for with a medical background, analysed all clinical trials
a younger, sexier, happier you” (Klatz & Goldman, on growth hormone to determine whether it could be
2003). used safely and effectively by healthy older patients.
“Sweet syringe of youth” (Langreth, 2000) . 63
They deliberately excluded studies in which the efficacy
of growth hormones was evaluated on the treatment
In 2003 Dr. Mary Lee Vance from the Medical of a specific disease. They conducted this research
department of the University of Virginia Medical Centre, because the use of growth hormone as an anti-ageing
published the article “Can growth hormone prevent agent is very controversial and yet is used as such by
many people, even though it has not been approved
63
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Chapter 10: Hormones: natural regulators 175

 for that purpose by the FDA, and its dissemination as officer found a thermos flask containing 26 ampoules
an anti-aging agent is therefore illegal in the US. in one of the swimmers’ bags. Thirteen of them were
The use of growth hormone as an anti-aging agent filled with a liquid. The other thirteen contained a
scores as one of the highest health-related searches powder described on the label as containing human
on the internet. According to Mary Lee Vance more somatrophin, i.e. human growth hormone. The female
than one and a half billion dollars’ worth of growth owner of the luggage told customs that the ampoules
hormone is sold every year, a third of which is belonged to her coach. The customs department were
probably not under prescription and therefore illegal. in no doubt, however, as to their purpose.
Proponents of the use of growth hormone as an anti-
aging agent claim that in 2002 more than one hundred
A CUSTOMS OFFICER DISCOVERS SOME AMPOULES....
thousand people obtained growth hormone without
NO SIR, THAT’S WATER FOR MY CONTACTS...
a prescription. Liu et al. conclude that there are few ...YOU KNOW... SWIMMING POOL, CHLORINE, EYES...
published data on the effect of growth hormone on the
elderly, but available evidence suggests that the risks
far outweigh the benefits if the hormone is used as
an anti-ageing agent in healthy elderly patients. The
most frequently occurring side effects were oedema in
soft tissue and joint pain, while few significant positive
effects on physical make-up were reported. In short,
there is little chance of growth hormone being made
available on the market as a legal anti-ageing agent.
The online search we conducted as a result soon
Growth hormone as performance-enhancing drug revealed why they were so convinced. We found a
The controversial fact that ‘fat tissue disappears and website64, which is now banned, containing a 9-page
muscle tissue appears’ when growth hormone is used manual for the “underground” user and which opened
has also made it irresistible to athletes since the early with a similar quote to that at the start of this chapter,
1980s and has led to illegal use. This became only too also from the Underground Steroid Handbook. The
obvious when in January 1998 newspapers reported last paragraph of the manual began as follows: “The
that the Chinese federation had pulled out a swimmer active substance somatrophin is available as a dry
and coach en route to the world championships in Perth. powder and has to be mixed with the solution in the
The team members’ bags were examined on arrival at accompanying ampoule before being injected.” There
Sydney airport during a routine control. The customs
64
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176 Part 3: Health has limits

is a fear, justified by this internet article, that this Movement Science and doping expert Harm Kuipers,
is just the tip of the iceberg of a widespread use of a former World Champion speed skater, says in
growth hormone as a performance-enhancing drug. this article that the WADA announcement is window
If this explains the phenomenal growth in demand, dressing to a large extent. The test only traces HGH
this wonderful medicine, one of the first products of and this disappears from the urine a few days after
modern biotechnology, will be seriously but unjustly administration. An “indirect” test on measurable, long-
discredited. term effects in the body after HGH use is much more
Even more distressing is the example in the previously necessary (as with EPO, see following paragraph).
mentioned article “Illegale hormonen”. According to Looking at the available studies, Kuipers doubts very
this article illegal growth hormone, extracted from the much whether HGH actually does anything, So he
pituitaries of deceased people, was on sale in the Dutch advised the WADA to investigate this first, and then, if
bodybuilders’ circuit back then. Bodybuilders who used the growth hormone does actually make a difference,
this product, of Russian origin, run the risk of getting to invest in an “indirect” test. Recently a review has
Creutzfeldt-Jakob disease in 10 to 20 years. The Dutch been written by scientists from the WADA and the
Health Inspectorate therefore issued a direct warning IOC (Barroso, Schamasch, & Rabin, 2009). From the
against the use of this illegal growth hormone. In our same year is the review Growth Hormone in Sport:
view it is unwise from a scientific perspective to use Beyond Beijing 2008 (Segura et al., 2009).
recombinant growth hormone without a prescription. So The importance of this topic is further stressed by
using this sort of illegal pituitary extract is tantamount the editorial “Game over for sports cheats” written
to playing Russian roulette. by Vicky Heath, the Chief Editor of Nature Reviews
There has been a urine test for tracing human growth Endocrinology (Heath, 2010). She welcomes the
hormone since 2004, which meant it was available “groundbreaking new initiative” that WADA recently
for the Olympic Games in Athens. The test was rolled out: the athlete biological passport. She writes:
developed by the German endocrinologist Prof. “in addition to the usual random drugs tests, athletes
Christian Strasburger, who has since improved it. are now required to undergo regular monitoring of
In response, the World Anti-Doping Agency (WADA) biological variables, such as their levels of hemoglobin
announced that athletes would be thoroughly tested or red blood cell count. Plotting these measurements
for performance enhancement with human growth over time and looking for abnormal variations should
hormone for the first time at the Olympic Games facilitate indirect detection of doping because the
in Beijing. The science section in the 26/27 July downstream effects often remain evident long after the
2008 edition of the Dutch newspaper NRC, was actual drug has disappeared from the body. On the face
largely dedicated to this doping control. Professor of of it, longitudinal screening is an excellent idea, but only

Chapter 10: Hormones: natural regulators 177

 time will tell whether the athlete biological passport scale for therapeutic treatments in very simple
proves an effective deterrent.” Like her we stress that “bioreactors”. In 1988 the American company
the potential for gene doping must be explored (see Amgen, based in California, brought recombinant
also next chapter on gene therapy) and we would like EPO produced in this way onto the market.
to end this section with an outcry from her, which we EPO is prescribed to patients with anaemia resulting
also fully endorse: “the use of performance-enhancing from kidney problems, and to cancer patients receiving
drugs is not restricted to elite athletes; published data chemotherapy. Before recombinant EPO came onto
suggest that abuse of anabolic androgenic steroids and the market, severe forms of anaemia were treated
other endocrine drugs is on the rise among high-school by blood transfusions with all the accompanying
and college athletes. Therefore, it is crucial that young limitations and risks (transfer of viral infections such
people and their mentors are properly educated about as AIDS or hepatitis B and C, and immune system
the risks involved. Clinicians and other health-care problems). The availability of recombinant human
professionals should take the initiative in this respect, EPO (r-huEPO) on the market has dramatically
as they have a duty of care to highlight the uncertain improved the treatment of anaemia and it is now one
benefits and potentially harmful effects of doping.” of the world’s best-selling medicines. The question is
whether it all finds its way to the real patients.
10.3. ERYTHROPOIETIN (EPO) For decades, top sportsmen and women have been
trying to boost their levels of red blood cells, because
“Did biotechnology ruin the Tour de France?” So it enables them to take up oxygen more easily. That’s
ran the headline of the article by Gaby van Caulil in the purpose of altitude training. The thin air stimulates
BIOnieuws of 8 August 1998 (Van Caulil, 1998). In the body to produce extra erythrocytes, enabling the
it he writes about the “dual” use of erythropoietin, sportsmen to perform better when they return to lower
or EPO. EPO is a hormone made by our kidneys, altitudes. That EPO is used as a blood doping agent is
which regulates the production of red blood cells a known fact and famous top sportsmen and women
(erythrocytes) in bone marrow. Red blood cells are have admitted taking it. Performance enhancements
packed with haemoglobin. This is the pigment that of approximately ten percent are possible. In top
gives blood its red colour and ensures that oxygen sports this is a monumental improvement. However,
is transported from the lungs to other bodily tissue. injecting EPO is dangerous. The hormone thickens
In 1985 molecular biologists inserted the gene that the blood, making it more difficult for the heart to pump
codes for human erythropoietin into the genetic this bodily fluid around the body. The sudden death
material of animal cells. These recombinant cells of 18 Dutch and Belgian cyclists between 1987 and
make it possible to produce human EPO on a large 1990 is still shrouded in mystery. These sportsmen

178 Part 3: Health has limits

died from unexplained cardiac arrest (some of them National Anti-doping Laboratory describe a more
in their sleep, when the heart rate is at its lowest). effective test (Lasne & de Ceaurriz, 2000). EPO is a
There are stories of cyclists having to get up every protein that consists of 165 interlinked amino acids.
two hours during the night to use the home trainer The amino acid chains in the recombinant EPO are
in order to keep their circulation going and thus stop identical in principle to that of the naturally occurring
their heart from giving out (Bloembergen, 2007). EPO. Yet there is a difference. These amino acid
And yet, EPO as a performance- chains have side chains that consist
ALTITUDE TRAINING...
enhancing drug is still popular of saccharide molecules (the chain
among sportsmen and women, CYCLING IS FUN... CYCLING IS FUN... is glycosylated). Different
because it remains difficult to CYCLING IS FUN... CYCLING IS FUN... saccharide chains are linked
trace with certainty. An indication to the natural EPO than to
of EPO use is the red blood the recombinant hormone. Taking
cell count. This is expressed as a this difference as a starting point
percentage of the total blood volume, the the French researchers designed
haematocrit value. A value of more than a detection method (Textbox 10.1).
50% suggests EPO use and the person It is, however, a labour-intensive and
in question is then suspected of doping. expensive way of tracing in multiple
The 1998 Tour de France has become stages (Van ‘t Hoog, 2008).
notorious because of the great many However, during the 2000 Olympic
participants suspected of using EPO, Games in Sydney this test was
determined on the basis of the used, in combination
haematocrit value. Up with an indirect blood
until 2000, however, test which was also
this suspicion was very published that same
hard to substantiate. year by Australian
Fortunately, strenuous researchers in the June
efforts were and still are being made edition of Haematologica. Rinze
to develop more watertight tests. A watertight EPO Benedictus (Benedictus, 2000) called it a “Sherlock
test will not only protect sportsmen and women from Holmes method”, whereby a number of different
the massive pressure of commerce, but also heal blood parameters are integrated in a model. By
the tarnished reputation of this great medicine. entering data from dozens of blood samples into the
In the journal Nature researchers from the French model, a mathematical representation can be built

Chapter 10: Hormones: natural regulators 179

 TEXTBOX 10.1. than positive charges. The effect of the voltage applied
The French EPO test. across the plate is to move both proteins downwards.
When the recombinant EPO reaches pH ~ 4.8 (~ pI),
The French EPO urine test uses electrophoresis, a there are as many negative as positive charges, and
chemical separation technique based on differences in the molecule ceases to move. That only happens
electrical charges. Urine samples are placed at the top with the natural EPO at pH ~ 4.2. There are several
on a plate of electrophoresis gel (figure 10.3) and a bands visible because natural and recombinant EPO
difference in voltage is applied across the plate. In the both have several (iso)forms with small reciprocal
figure the cathode (negative) is at the top and the anode differences (they are microheterogenic). The bands
(positive) at the bottom. A gradient in pH is also applied are made visible using two dyeing methods (double
across the plate; from top to bottom, the pH lowers and immuno-blotting and chemoluminescence). This
it therefore becomes more acidic. Proteins without side ‘double blot’ procedure makes the test unique and
chains have positive and negative charges. The more minimises the chance of false positive results (Coons,
acidic the more positive charges, and the less acidic 2004).
the more negative. At a certain pH, there are exactly
the same number of negative as positive charges and
the protein is electrically neutral. This is called the iso-
electric point (pI).
Since the recombinant and natural EPO have the
same amino acid chain in principle, they can’t be
differentiated in this way. The side chains, however,
consist of different sugar molecules which have a
negative charge at a higher pH. This means that for a
certain pH (e.g. 5), the balance between negative and a b c d e f g h
positive charges is different for recombinant and natural
EPO. The sugar side chains of the natural EPO have Figure 10.3. EPO test using electrophoresis (from Lasne
relatively more negative charges than the recombinant and Ceaurritz, 2000; adapted); a. purified from urine,
EPO, in other words, the pI of natural EPO is lower commercially available; natural human EPO; b. commercial
than that of the recombinant EPO, respectively in the recombinant EPO-β; c. commercial recombinant EPO-α; d.
range 3.9 - 4.4 and 4.4 - 5.1. EPO samples are brought urine from a control person; e. and f. urine from two anaemic
up to the negative upper side at pH = 5.2, i.e. both patients treated with recombinant EPO-β; g. and h. urine from
natural and recombinant EPO have more negative two ‘positive’ cyclists from the 1998 Tour.

180 Part 3: Health has limits

up so that with the right threshold value it selects (Textbox 10.2). The 2007 Tour was also seriously
EPO users and doesn’t give false positives. There tarnished. For the first time in history a leader and
are two versions of the model: one that predicts wearer of the yellow jersey was disqualified from
use up to three weeks before the test, and one the course. In 2008 the Tour was hit on the first day
that gives a positive result for doping a few days and after the first week two “positive” cyclists had
before. Internist and EPO expert J. Marx, of the already been sent home by their teams. A week later
University Medical Center Utrecht, said at the time the same occurred for a two-times stage winner. The
that this theoretically signalled the end of EPO as editor of de Volkskrant (“Any hope of clean Tour is
an undetectable performance-enhancing drug. “But again lost”) on 19 July 2008 wrote that in the case of
because this indirect, Australian model will probably the first two cyclists we could still console ourselves
not stand up to legal scrutiny, the French urine test with the thought that neither had a place in the overall
still has to be carried out after the blood test. Then ranking. But for the latter case it was a completely
the sportsman or woman has absolutely no chance”, different matter:
says the EPO expert in the article by Benedictus. “Now that one of the most promising young cyclists
After Sydney 2000 the WADA decided that the has succumbed to the temptation, the link between
French urine test alone was reliable enough to doping and cycling has become all too clear. ….It is
sanction a sportsman or woman caught using EPO as true as it is impotent to conclude that the latest
(Köhler, 2008). Backed by the IOC, which used to doping affair has brought the credibility of cycling in
draw up the doping list and approved the doping general and of the Tour in particular to a new low
labs, WADA is the highest doping authority in the point. Because how is it possible to restore credibility?
world. Thus with the EPO urine test WADA opted for Stricter controls are only part of the solution, if only
a test that (directly) shows the EPO molecule itself. because the limits of what is still defensible on legal
The results of EPO use are visible in blood, but that is and human grounds are becoming visible.”
an indirect test. Just as Marx expected, indirect tests At least with a really watertight test the legal
are difficult from a legal standpoint. Like Benedictus, possibilities improve. The criticism of the use of
Köhler uses detective work as a metaphor: there is the direct urine test alone is gaining momentum, as
a body, and probably a murderer too, but no murder demonstrated in an interview with the doping expert
weapon. In 2004 the urine test was validated and Rasmus Damsgaard (Randewijk, 2008). There
every year we hear the same cry yet again: this will are also increasing calls for a combined direct and
be the cleanest Tour ever! So far, however, this has indirect test (Köhler, 2008). The above-mentioned
proved to be anything but the case. In 2006 the Tour Harm Kuipers says in this latter newspaper article
was a complete disaster in terms of doping scandals that the international skating union has been

Chapter 10: Hormones: natural regulators 181

 TEXTBOX 10.2. place in the Tour of 2006. Most of the Spanish cyclists
Operación Puerto file still open. had a lucky escape. …The file remains open. …”
One of the statements made by Jaksche in the interview
This was the headline to an intermezzo in an article on the subject reads as follows: “The doping problem is
entitled “Doping as perpetual motion” by Marije not my problem, or that of Basso or Ulrich, it is a problem
Randewijk in the Dutch Volkskrant of 12 July 2008. It of the system. We are all victims and perpetrators alike.
is an interview with Jörg Jaksche, one of the leading You are forced to go along with it, and because you do
players in Operación Puerto: it, you force others to come along with you. It’s perpetual
“On 23 May 2006, following a raid by the Guardia Civil motion.”
on the laboratory of haematologist Merino Batres and In the first week of October 2008, the Spanish courts
the Madrid apartments of Dr. Eufemiano Fuentes, 220 dismissed the investigation into this biggest ever doping
blood bags, growth hormones, anabolic steroids and scandal, thereby permanently closing the Operación
EPO were seized. On the same day Fuentes and Manolo Puerto file from a legal point of view. “Only the sport of
Salz, team leader of Liberty Seguros, were arrested. … cycling refuses to close the book on ‘Puerto’” according
Three more key figures in the biggest doping affair in the to an article in the NRC of 4 October 2008. “It’s better to
sport were picked up. …The collaboration with Fuentes randomly search for perpetrators than act as if nothing
cost top favourites Ivan Basso and Jan Ullrich their is going on.”

combining blood tests with urine tests since 2000. to the researchers the levels in the milk were so
The main problem with the urine test on its own is high and stable that they could be used in clinical
that there are an increasing number of variants of experiments. The intention is, amongst other things,
recombinant EPO on the market, which look more to incorporate the EPO in medicines for patients with
and more like the natural EPO, particularly because cardiovascular disorders. Its use in the sporting world
human tissue rather than animal cells is now used for is obviously not mentioned.
production. Unfortunately, the latest developments In order to increase production still further a number of
in the area of modern biotechnology are gratefully pigs have also now been modified and the hope is to
used in many illegal laboratories. In the Korea Times obtain more EPO from pig milk, if possible at the lowest
of 22 February 2006 there was a report on one possible cost price. Whatever happens, it is virtually
such development. A team of researchers from the impossible to trace all new types that come on the black
University of Gyeongsang in South Korea announced market. According to Damsgaard the only solution is
that they had genetically modified nine mice in such a to develop a benchmark of what a normal test result
way that they produced EPO in their milk. According should be for an individual sportsman or woman.

182 Part 3: Health has limits

 THE UPSCALING OF EPO-PRODUCTION is a sign that the drug-plagued sport is turning a corner
USING PIGS INSTEAD OF MICE should think again. At a race for cycling’s budding
stars several weeks after the 2009 Tour, no positive
MICE JUST WEREN’T DOING doping tests turned up either—until customs officials
IT FOR ME ANYMORE! raided a Ukrainian team bus, seized doping gear, and
investigators later wrested admissions of blood doping
and use of endurance booster EPO during the event.
The 2010 Tour began on Saturday 3 July in Rotterdam,
with cycling bosses holding their breath in the hope that
the race would be clean—and not just in appearance.”
Now three weeks later the Tour has a winner again, the
same as last year, i.e. the Spaniard Alberto Contador.
Will it again be a Tour free of positive doping tests? For
that we have to wait another couple of weeks for the
results, too late probably for this book. But if so, does
that mean then that Pandora’s hope is lifted from the
But is a watertight test the end of the story? Probably bottom of the box, or does it herald the new era of gene
not. Researchers who have inserted the gene for EPO doping? Let’s hope not!
in mouse muscles with the intention of developing In conclusion, as far as EPO as a medicine is
therapeutic applications, are afraid that once their concerned, there may well be more areas of application
research delivers results, sportsmen and women in the future. In 2003 German researchers came to the
will also use this gene therapy for doping (see also conclusion that EPO is also a good candidate for the
Chapter 11 on gene therapy). The ethical question is treatment of schizophrenia. The hormone would not so
whether this is reason enough to halt such research, much tackle the symptoms (paranoia and delusions)
which could be life-saving for some. as the underlying neurodegeneration, which proceeds
On 3 July 2010 Jamey Keaton, Associate Press Writer despite successful medication. A clinical trial has begun
published the article “Doping lurks as wild card at 2010 in eight German centres. The same researchers had
Tour.” 65
He writes: “Last year’s Tour de France was already observed the beneficial effects of EPO in 2002
notable for more than just Lance Armstrong’s return. It in the repair of a brain haemorrhage. The status of this
was also free of positive doping tests. This came after research can be found on the website of the group.66
three straight years during which cycling’s main event
was marred by drug cheats. But those who believe this
65
sports.yahoo.com/sc/news?slug=ap-tourdefrance-doping

Chapter 10: Hormones: natural regulators 183

 10.4. PUREGON™: FOLLICLE-STIMULATING appears to be a rich source of two other members
HORMONE (FSH) of the family of fertility hormones: follicle-stimulating
hormone (FSH) and luteinising hormone (LH).
Sooner or later in their lives most adults want to These are the active components of the preparation
have children. Those who fail in their attempts to Humegon® which stimulates follicle ripening.
fulfil this desire often feel frustrated, desperate Thanks in part to the advent of in-vitro fertilisation
and hopeless, with all the ensuing consequences. (IVF), in which both the above-mentioned
In our Western society about 15% of all couples preparations are used, the collection of urine
have fertility problems. Fertility hormones have samples in the Netherlands and beyond has risen to
been prescribed for decades to solve this issue. millions of litres per year. The collection of this special
The prescription of the fertility hormone FSH made urine has always been a delicate matter because
from genetically modified animal cells is fairly new. it requires the cooperation of volunteers. In less
These cells are originally isolated from the ovary of than 80 years this urine collection has undergone
a Chinese hamster and can then be grown in flasks a veritable metamorphosis. First it was collected by
and bioreactors. bike in the town of Oss. Then a regional collection
In 1923 Organon, once part of Akzo Nobel but since was conducted by horse and cart. After the Second
10 March 2009 part of the American pharmaceutical World War the operation was expanded to the whole
company Merck, began producing insulin under the country by car. The organisation ‘Moeders voor
licence of the University of Toronto. Zwanenberg Moeders’ (Mothers for Mothers) now has a whole
Vleesbedrijven, a meat company, supplied its fleet of ten tonne trucks for the job.67
subsidiary Organon with “worthless” slaughter
waste - in this case pancreas - from which Organon
THE COLLECTION OF URINE FROM
PREGNANT WOMEN IS VERY SUCCESSFUL!
isolated the valuable insulin. Less than a decade
later, this approach still appeared to be a golden
USE ONE OF THE TOILET BOOTHS, MADAM...
formula; in 1932 Organon began with the isolation of
the valuable fertility hormone hCG (human Chorionic
Gonadotrophin) from the “waste” urine of pregnant NO DRINKIN
G
women. The hormone which induces ovulation was WATER!

sold under the brand name Pregnyl® and was used

in fertility treatment. This would later be the scenario
for the waste urine of postmenopausal women. This

66
www.neuroprotection-schizophrenia.de 67
www.moedersvoormoeders.nl

184 Part 3: Health has limits

Fertility hormones are extracted from urine using is encoded not by one but by two genes. Saccharide
classical biochemical methods. The foremost groups then have to be linked to the protein chains
complications are the variable quality of the urine, (a process called glycosylation) if the FSH is to
the large scale of production and the strict purity be activated (Figure 10.4). Genetically modified
criteria which the injection preparations have to microorganisms seem unable to glycosylate. In
satisfy. Reasons enough for Organon to conduct a contrast, after genetic modification, Chinese hamster
search in the 1980s for an alternative production cells seemed perfectly capable of performing
method using modern biotechnology. In July 1996 the whole process; moreover, they are safe and
they hit the jackpot. Organon came on the market widely accepted for the preparation of recombinant
with human FSH (Puregon™), made with genetically medicines.
modified Chinese hamster cells. Since Puregon is more than 99% pure, it can be
FSH is one of the most complex protein molecules in administered to patients subcutaneously rather than
the human body. It consists of two protein chains and intramuscularly. Patients can also inject themselves.

Follicle Stimulating Hormone (FSH) is composed of two protein chains (α and β) and contains a number of sugar groups.
For the production of FSH the Chinese Hamster Ovary (CHO) cell line is used.

The human FSH gene is Chinese Hamster Ovary (CHO cell)

built into the DNA of the
Chinese Hamster cell line
mRNA

Transcription
Human DNA Complete FSH

Translation

DNA

Nucleus

The CHO cell assembles

the α- and β-chains,
couples sugar groups
to it and excretes FSH

Figure 10.4. Mammalian cells for FSH production, adapted from Olijve & Houwink (1993).

Chapter 10: Hormones: natural regulators 185

 The recombinant FSH is homogenous and contains no available in recombinant form and are approaching
human protein impurities, as is the case with traditional practical application.68, 69, 70
preparations. This makes it ideal for use in patients who
are allergic to FSH from urine and thus have to cease 10.5. IN CONCLUSION
further treatment. Because the manufacture of Puregon
does not rely on the availability of the raw ingredient In this chapter we have taken a close look at three
(urine), production is more flexible. In short, a great hormones made with the help of modern biotechnology
product, a fantastic production process, and fortunately and sold as medicines. Two of them save lives and the
no incorrect applications (at least as far as we know). third creates lives. All three are really wonderful, pure
So has the organisation “Mothers for Mothers” become medicines with many benefits compared to the old
obsolete? Not quite yet. As long as there are no better drugs. Yet two of the three have a tainted reputation,
alternatives for the other fertility hormones that are also mainly because of their illegal use as performance-
obtained from urine, this organisation will still play a enhancing drugs. That’s a crying shame, especially
vital role. However, it is highly likely that sooner or later since it is precisely these two that are often used as
modern biotechnology will come up with alternatives life-saving treatments in patients. Nevertheless, they
for these also. In fact it has almost reached that point. are still beautiful examples of very worthwhile products
Both the two other mentioned fertility hormones are of modern biotechnology.

68
www2.cochrane.org/reviews/en/ab005070.html
69
www.gfmer.ch/Endo/PGC_network/Recombinant_luteinizing_hormone_Pou.htm
70
www.fertstert.org/article/S0015-0282(09)00502-0/abstract

186 Part 3: Health has limits

 10.6. SOURCES Liu, H., Bravata, D. M., Olkin, I., Nayak, S., Roberts, B.,
Garber, A. M., et al. (2007). Systematic review: The
Barroso, O., Schamasch, P., & Rabin, O. (2009). Detection safety and efficacy of growth hormone in the healthy
of GH abuse in sport: Past, present and future. Growth elderly. Annals of Internal Medicine, 146(2), 104-115.
Hormone & LGF Research, 19(4), 369-374. Olijve, W., & Houwink, E. H. (1993, October). Biotechnologie
Benedictus, R. (2000, 16 September). EPO-doping in toom. vernieuwt geneesmiddelen. Chemisch Magazine.
Bionieuws. Pownall, M. (1994). Biotechnology triumph brings hope to
Bloembergen, J. (2007, 7/8 July). Limonade in de benen. NRC. more patients. International Biotechnology Laboratory
Coons, R. (2004, 1 November). New detection methods help (May).
level the field. Genetic Engineering News. Pramik, M. J. (1999, 1 January). Recombinant human growth
Elkins, R. (1999). HGH: Age-Reversing Miracle. Orem, UT, hormone: one of biotech’s first products expands
Woodland Publishing. indications. Genetic Engineering News.
Gilbert, E. M. D., Jamie, J.J., & Gross, S. (1999). Staying Randewijk, M. (2008, 19 July). Dopingexpert juicht epo-Tour
young: growth hormone and other natural strategies toe; interview met Rasmus Damsgaard. De Volkskrant.
to reverse the aging process. Boca Raton, FL, Age Rudman, D., Feller, A. G., Nagraj, H. S., Gergans, G. A.,
Reversal Press. Lalitha, P. Y., Goldberg, A. F., et al. (1990). Effects of
Heath, V. (2010). Game over for sports cheats? Nature human growth-hormone in men over 60 years old. New
Reviews Endocrinology, 6(8), 413. England Journal of Medicine, 323(1), 1-6.
Klatz, R., & Goldman, R. (2003). The new anti-aging Segura, J., Gutiérrez-Gallego, R., Ventura, R., Pascual, J.,
revolution: stopping the clock for a younger, sexier, Bosch, J., Such-Sanmartín, G., et al. (2009). Growth
happier you! (3rd ed.). Laguna Beach, CA, Basic hormone in sport: beyond Beijing 2008. Therapeutic
Health Publications, Inc. Drug Monitoring, 31(1), 3-13.
Klatz, R., & Kahn, C. (1998). Grow Young with HGH: The Van ‘t Hoog, A. (2008, 16 August). Scheidsrechter met kolom
Amazing Medically Proven Plan to Reverse Aging. en massaspectrum. C2W.
New York, Collins. Van Caulil, G. (1998, 8 August). Heeft biotechnologie de Tour
Köhler, W. (2008, 26/27 July). Epodope. NRC. de France bedorven? BIOnieuws.
Langreth, R. (2000). Sweet syringe of youth. Forbes Global. Vance, M. L. (2003). Retrospective - Can growth hormone
Lasne, F., & de Ceaurriz, J. (2000). Recombinant prevent aging? New England Journal of Medicine,
erythropoietin in urine. Nature, 405(6787), 635. 348(9), 779-780.

Chapter 10: Hormones: natural regulators 187

11 GENE THERAPY:
A PANACEA FOR GENETIC ABNORMALITIES?

“I just bumped into a man who we admitted to the Antonie van Leeuwenhoek hospital four years ago. I can
attribute the fact that he is now in good health due to the gene therapy he received back then.”

Winald Gerritsen, director of the Cancer Centre in the Free University Amsterdam Medical Centre, March 1999.

The January 1996 issue of Chemisch Magazine contained an article in the New Technological Trends of the 21st
century section entitled “Gene therapy causes fourth medical revolution.” The article begins as follows:
“More than 4,000 diseases and abnormalities are caused by a defect in a single gene. Although still in its infancy,
gene therapy may be the solution. Insiders believe that this therapy will result in a new revolution in the medical
world in the 21st century. History has already witnessed three major turnarounds in the fight against disease. The
first was with a greater focus on sanitation facilities and sewers, which went a long way to suppressing infectious
diseases.” - We have already read about Louis Pasteur’s pioneering activities in the area of hygiene in the chapter
on wine. – “Then came anaesthesia, which enabled doctors to treat patients under sedation. Finally, there was the
introduction of vaccines and antibiotics, which prevented and treated many viral and bacterial infectious diseases.”
Now, more than a decade later, we will address the following question in this chapter: Does gene therapy really
herald the fourth revolution? First, however, we will look at what gene therapy actually involves.

THE FOUR MEDICAL REVOLUTIONS

THE SEWER THE SEDATION THE ANTIBIOTICS THE GENE THERAPY?

J. Tramper, Y. Zhu, Modern Biotechnology – Panacea or new Pandora’s box? 189

DOI 10.3920/978-90-8686-725-7_11, © Wageningen Academic Publishers 2011
 11.1. WHAT IS GENE THERAPY?

The idea behind gene therapy is simple. When a

child is born with a serious disease caused by a
gene defect, it is possible in theory to provide the
diseased cells with a normal copy of the gene,
thereby removing the defect. The practice, however,
is far from simple. Whilst it is true that medical and
DNA
biotechnological researchers have developed a few
techniques to insert ‘corrective genes’ in (diseased)
cells, stable integration of a ‘working corrective gene’
in the DNA of the cells is another matter entirely.
The simplest method is to inject naked DNA,
containing the corrective gene, into tissue. Nowadays
a DNA molecule is easy to make and to buy. However,
in contrast to other cell types, only muscle cells Figure 11.1. Liposome with corrective DNA.
can take up loose DNA. A more efficient procedure
is to use a so-called vector, such as a liposome The most efficient method is the use of viruses as a
or virus. A liposome is an artificially synthesised vector. The advantage of viruses is that they have a
bubble membrane containing, for instance, an natural tendency to stick to host cells and are able to
aqueous solution with DNA (Figure 11.1). The bubble inject their genetic material into the host cells. This is
membrane can be made up of a double layer of their way of multiplying and “surviving”. The virus DNA
molecules that resemble phospholipids, the natural then becomes integrated in the cell DNA, causing
molecules that are the main component of the cell the cell to make large quantities of the new virus and
membrane in living organisms. The molecules have become “sick”. Viruses are the Trojan horses of biology:
a hydrophilic (water-loving) head and a hydrophobic they are experts at incorporating foreign DNA.
(water-repellent) lipid tail. In theory, liposomes can So a virus is a perfected instrument for introducing
be used to take their content into individual cells genes into cells. With the help of recombinant DNA
by allowing the membrane to fuse with the cell techniques it is possible to replace the genes of a virus
membrane. If the content were to be, for example, that are crucial for replication of the virus with human
the corrective DNA, this could be transferred via the genes. The virus is consequently weakened so much
liposomes into the target cell. that it can no longer kill our cells, but is still able to deliver

190 Part 3: Health has limits

DNA to the cell. Such viruses are obviously first tested in gene therapy on patients. Gene therapy has been used,
animals to highlight any of the possible major problems: for instance, as the last resort in the case of children
with serious and untreatable congenital disorders.
• It is difficult to infect a large number of body cells So that it can be used in gene therapy, a corrective
simultaneously with weakened viruses. gene (or genes) must be incorporated in the virus DNA
• If the infection succeeds, the genes often fail to reach in such a way that the viruses can no longer replicate
the cell nucleus which contains the cell’s DNA. in a host cell and so can no longer make this cell sick.
• If they do reach it, they become incorporated in Many viruses infect a variety of cell types, while gene
abnormal locations in the DNA, often causing them therapy is directed at specific cells/tissues. As of this
to be ineffective. moment, there are two principal ways of using virus
• If, in the beginning, the genes appear to be working vectors for gene therapy to reach this goal. The most
reasonably well, this action usually stops after a few common technique involves a doctor removing cells
weeks. containing the defective gene from the patient’s body,
adding a corrective gene to these cells in the laboratory
Despite the disappointing results in animals, medical using a genetically modified virus and then implanting
professionals have still been relatively quick to test the cells in the patient’s body again (Figure 11.2).

In the test tube

Virus infects cell

Genetically Integrate
modified virus Placing
Cytoplasm
Cell nucleus back

Body cells
of patient

Production
Body cells ‘corrective’ Cell membrane
of patient enzyme

Figure 11.2. Principle of gene therapy with a genetically modified virus.

Chapter 11: Gene therapy: a panacea for genetic abnormalities? 191

 Initially it was mainly blood cells that were ‘corrected’, were transferred. Details can be found on their very
but these have a limited life span and so ‘the cure’ informative and user-friendly website71. In June 2010,
is also of short duration. As a result current research for example, it showed that the US (64.3%) and
now focuses on bone marrow cells, which continue to Europe (29.3%) had carried out the greatest number
divide and ensure the creation of blood cells. of trials by far; the Netherlands came in at 1.6% with
In the second technique doctors directly administer 27 trials. Cancer treatment outscored the rest with
the virus vector with the corrective gene to the tissue 64.5%. The great majority (60.5%) of all clinical trials
manifesting the gene deficiency. This approach, were still in Phase I (see Textbox 9.1. Phases of drug
therefore, is only effective against local abnormalities, development). In short, this is a very rich source of
such as cystic fibrosis, or inherited muscular disorders up-to-date information about gene therapy. Also a rich
like Duchenne muscular dystrophy. In principle, tumours source of rather up-to-date information is the website72
can also be treated in this way by administering a virus of the Human Genome Program sponsored by the
vector with a “suicide” gene; then, when the tumour US Department of Energy Office of Science, Office of
cell is treated with certain chemotherapies, this gene Biological & Environmental Research.
ensures that the tumour cell self-destructs. The concept of gene therapy has existed for quite a
while. In 1972 Professor Theodore Friedmann and
11.2. A SHORT HISTORY OF GENE THERAPY his colleague Richard Roblin, from the University of
California in La Jolla, wrote in the leading scientific
Since 1997 several medical researchers and an editor journal Science about the possibilities of gene therapy
of a medical journal have been keeping a database in genetic abnormalities (Friedmann & Roblin, 1972);
on clinical trials for gene therapy (Edelstein, Abedi, & Friedmann was involved from the very beginning of
Wixon, 2007; Edelstein, Abedi, Wixon, & Edelstein, gene therapy. In 1989 researchers at the American
2004). They obtain the data from official bodies, from National Cancer Institute in Bethesda were the first to
literature, at conferences and directly from researchers experiment with gene therapy in humans (Rosenberg
or research sponsors. By June 2010 the database et al., 1990). Five patients with advanced stage cancer
contained almost 1,650 trials, some finished, some were involved in this pioneering trial. The trial showed
ongoing and a few with “start-up authorisation”. An that gene therapy in principle also works in people.
analysis is made continuously not only of numbers It also revealed some important prerequisites for
and geographical distribution but also of the medical subsequent clinical studies with gene therapy. After
reasons behind the trials and the way in which genes that, the next approved clinical trial took place in 1990

71
www.wiley.co.uk/genmed/clinical
72
www.ornl.gov/sci/techresources/Human_Genome/medicine/genetherapy.shtml

192 Part 3: Health has limits

(Blaese et al., 1995). For the first time some success The mood changed, however, at the end of the 1990s
was achieved with gene therapy carried out in two girls following two serious events. The first took place
with SCID (Severe Combined Immune Deficiency) - a on 17 September 1999 when the 18-year old Jesse
very serious congenital syndrome in which the immune Gelsinger died as a result of a gene therapy treatment
system doesn’t work; however, Section 11.3 describes (Raper et al., 2003). The death was attributed to a
how, ten years later, the treatment of SCID with gene totally unexpected and devastating inflammatory
therapy had disastrous results. From 1990 to 1999 response to the adenovirus used (Textbox 11.1). The
the number of clinical trials rose dramatically (Figure FDA therefore stopped this and a few other trials.
11.3). In general, expectations ran high in this period, In February 2005 the American Ministry of Justice
but there were still voices of concern to be heard about delivered its final verdict on this case (Couzin & Kaiser,
the possible risks of gene therapy, and some critics 2005). The University of Pennsylvania, where the trial
pointed out that this treatment had thus far delivered had been carried out, was held responsible and had
little in the way of therapeutic benefits. to pay a settlement fee of $517,000. In addition, extra

132 restrictions on gene therapy research were imposed

on the doctors who had performed the therapy.
116 117 116
112 In 2000 morale was boosted somewhat by a report
108

96
101
from France about successful gene therapy treatment
95

85
89 in ten children with a rare form of SCID (Cavazzana-
82
76 Calvo et al., 2000). However, the joyous enthusiasm
67 68 didn’t last longer than two years, when at the end of
2002 there was an alarming report that the two children
51
in which the greatest number of “corrective” cells had
37 38
been implanted had developed leukaemia as a side
23 effect. The study was voluntarily stopped and, once the

8
14
protocol was revised, it was restarted with lower doses
1 2 of corrective cells. In Section 11.3 we elaborate on this
20 6

09
20 0

20 4

20 8
20 2
8

20 7
96

99
90

issue.
20 3

05
89

98
94
92

10
01
97
93

95
91

0
0
0

0
0
0
98

20

20

20

20
19
19
19
19
19
19
19
19
19
19
19
≤1

Figure 11.3. Number of approved clinical trials over the years The death of Jesse Gelsinger and the serious
(last updated October 2010 ). 73
consequences thereof overshadowed a positive result
of gene therapy reported at the time. At the end of 1999
the company Avigen declared that the first patients
73
www.wiley.co.uk/genmed/clinical/

Chapter 11: Gene therapy: a panacea for genetic abnormalities? 193

 TEXTBOX 11.1. resistance response. This resistance destroys the
The Gelsinger case. cells that are infected with the virus and it is precisely
these cells that should set the gene therapy in motion.
A virus is a packaged piece of DNA, that specialises However, gene therapy was and still is a spectacular,
in offering up its gene package to host cells. For new, experimental therapy and successes in this
example, the adenovirus, which is responsible for area are bound to attract attention. Such it was that
some of our colds, can bind to almost any cell type Jessie Gelsinger, who when he turned eighteen had
and then insert its DNA into the host cells. One a serious, but not life-threatening metabolic disease,
such adenovirus, genetically modified with a view to was injected with an enormous quantity of modified
gene therapy, appeared to work well in animal tests, adenovirus with the aim of correcting his metabolic
but in practice turned out to be unsuitable for gene disease. Four days later, Jessie died of a massive
therapy. The DNA of this virus is not built into our untreatable immune response to the weakened
chromosomes. It therefore only cures as long as the adenovirus. In one of his columns from 2001 about
virus multiplies in our cells and that usually stops after gene therapy, Emeritus Professor Piet Borst, scientist,
a few weeks have passed. Additionally, there is also columnist and long-time director of the Dutch Cancer
an immunological complication. People usually have Institute in Amsterdam, argued that the gene therapy
antibodies protecting them against adenoviruses and performed on Jessie Gelsinger was medically and
therefore it is necessary to inject huge quantities scientifically irresponsible. He concluded (and we
of the virus. That is a risky practice in patients with totally agree with him on this point): “This is not how
a compromised immune system. Moreover, the it should be done!” After this unfortunate incident the
weakened virus is often still able to generate a powerful rules in America have obviously been tightened.

with haemophilia B were experiencing positive effects from the blood of seropositive donors. At the time of
of their experimental gene therapy. Haemophilia B is a Avigen’s report, the first three patients, who by the
rare blood-clotting disorder. Because of a congenital end of 1999 had already received experimental gene
gene defect the bodies of sufferers from this disease therapy some months before, were making factor IX
don’t make any coagulation factor IX. Regular injections themselves again, albeit in low concentrations. Avigen
with coagulation factor IX from donor blood prevents inserted the gene for factor IX in an adeno-associated
them becoming handicapped or dying of internal virus (AAV) (Figure 11.4). Encouraged by the positive
(mainly in the joints) or external haemorrhaging. In results, Avigen is sponsoring clinical trials at Stanford
the 1980s many of these patients contracted AIDS, University Medical Center and the paediatric hospital
because their coagulation factor preparation was taken in Philadelphia (Sedlak, 2003).

194 Part 3: Health has limits

1. Original virus DNA is being removed of vascular medicine and a specialist
in the genetics of lipid metabolism at
Factor IX gene
the University of Amsterdam and his
colleague Erik Stroes, investigated
whether it was possible to cure a
2. Gene for factor IX is being implanted in the
virus mantel of adeno-associated virus (AAV) rare lipid metabolic disorder with gene
therapy using AAV. About 30 people in
the Netherlands have a genetic defect
whereby lipoprotein-lipase (LPL)
is poorly made or not made at all in
3. Modified virus is being injected their bodies. LPL is a fat-processing
in muscle tissue of the patient
4. Virus penetrates cell enzyme that splits fatty acids from the
Virus implants DNA into cell nucleus fat in the lipoprotein. In these 30 people,
however, the lipoproteins in the form of
big fat balls (chylomicrons) continue to
circulate in the blood. They end up in
the pancreas, where they can cause a
very painful infection. A strict fat-free
and alcohol-free diet is currently
5. Cell produces factor IX the only thing that helps a little.
Kastelein and Stroes incorporated
6. Factor IX enters bloodstream of patient the LPL gene into an AAV and, following
and restores coagulation capacity of the blood successful animal trials, they began in
2005 with clinical trials on patients. The
trial involved eight patients, each of whom had suffered
Figure 11.4. Gene therapy to treat haemophilia B, adapted from a pancreatic infection more than five times. In
from Köhler (2000). June 2007 Stroes was able to present the first hopeful,
provisional results in Seattle at the annual congress of
AAV is like a cold virus, but without the adverse effects. gene therapists.
The immune reaction is minimal and the location where the He reported that the pain after a great many injections
gene is incorporated is well-known. The risk of it activating under sedation into both thighs was minimal, as were
a cancer gene is also slight. John Kastelein, a professor the side effects. The fat content in the blood fell and the

Chapter 11: Gene therapy: a panacea for genetic abnormalities? 195

 effect appeared to be long-lasting. Some of the patients and obesity. Professor Kastelein was in 1998 one of
also reported having fewer stomach aches. This first the founders of Amsterdam Molecular Therapeutics Inc.
trial was to demonstrate safety. Now the correct dosage (AMT), a gene therapy company based on the concept
must be found. At the very least this early success gives of gene replacement in hereditary lipoprotein disorders
hope to around 5,000 of these patients worldwide; but it (Textbox 11.2).
may also be important for other, much more frequently
occurring disorders in which lipid metabolism plays a Another provisional success story was published in April
role, for example, cardiovascular disorders, diabetes 2008 by a British/US research group in the online edition

TEXTBOX 11.2. from an ongoing Canadian clinical study indicate that

Glybera: gene therapy for lipoprotein-lipase a single administration of Glybera in LPLD patients
deficient patients .74
results in a remarkable improvement in the ability to
break down the chylomicrons that transport dietary
Lipoprotein-lipase-deficiency (LPLD) is a seriously fat (triglycerides). LPLD patients are incapable of
debilitating, and potentially lethal, orphan disease, for clearing chylomicrons which are responsible for
which no approved therapy exists today. The disease causing significant morbidity and mortality. “The long-
is caused by mutations in the LPL gene, resulting in term improvement in chylomicron handling following
highly decreased or absent lipoprotein-lipase (LPL) Glybera administration is very impressive”, said Dr.
activity in patients. LPL activity is needed in order to André Carpentier, co-investigator from the University
break down chylomicrons, large fat-carrying particles of Sherbrooke, Quebec, Canada, who designed and
that are formed in the gut and enter the circulation after analysed the chylomicron sub-study. “These data
each meal. When such particles are not adequately are important, because the major complications
broken down they accumulate in the blood, and they observed in LPLD patients, including pancreatitis, are
may obstruct small blood vessels, which in turn can a consequence of chylomicron overload. They also
lead to pancreatitis. Recurrent pancreatitis in LPLD constitute evidence for a long-term clinically relevant
patients can result in difficult-to-treat diabetes. On lipoprotein-lipase activity induced by Glybera” noted
June 4, 2010 Amsterdam Molecular Therapeutics the principal investigator, Prof. Daniel Gaudet, from the
reported new data showing that its lead product University of Montreal, and ECOGENE-21 clinical study
Glybera results in the break-down of chylomicrons center, Chicoutimi, Quebec, Canada. These new data
in LPLD patients. Glybera is a gene therapy product provide a basis for explaining the mechanism of action
that induces functional lipoprotein activity. New data of Glybera in LPLD patients, and in general for continued
pharmacologic activity after one-time gene therapy.
74
www.amtbiopharma.com

196 Part 3: Health has limits

of The New England Journal of Medicine (Bainbridge neck cancer. This product, Gendicine, was approved by
et al., 2008). Six patients between 17 and 26 years old, the Chinese authorities on 16 October 2003, after more
who had gone virtually blind because of a rare congenital than five years of clinical trials. The article describes not
disease, regained a little of their sight following gene only the activities that led to the successful market entry
therapy. The improvement in the 18-year old British man, of Gendicine, but also the educational campaign to inform
Steven Howarth, was particularly spectacular. Here too, the public, the building of the production facility and the
the trial was carried out first to demonstrate safety. The technology and quality controls used to guarantee the
researchers now want to treat children whose sight has production of a safe and effective product. The Chinese
not deteriorated as much. Trials in young dogs have government’s policy of heavily promoting R&D in the
actually demonstrated a much more drastic improvement area of gene therapy is also emphasised in the article.
in sight than in adults treated thus far.
On 3 June 2010 scientists from Mount Sinai School of 11.3. SCID CHILDREN
Medicine in New York reported to the press (United Press
International) that they had developed a gene therapy In 1990 the first partially successful gene therapy
that is safe and effective in reversing advanced heart procedure was carried out in two girls with a specific
failure. The researchers said the therapy, called Mydicar, form of SCID (Blaese et al., 1995). SCID is an
is designed to stimulate production of an enzyme that abbreviation for Severe Combined Immune Deficiency.
enables the failing heart to pump more effectively. In a The syndrome consists of several very serious
Phase II study, injection of the gene SERCA2a through congenital disorders, resulting in a very fragile immune
a routine, minimally invasive cardiac catheterisation system. Without treatment, patients often die before
was safe and showed clinical benefit in treating and they reach the age of one or two. In these patients the
decreasing the severity of heart failure. The data indicate enzyme adenosine deaminase (ADA) was missing, or
that SERCA2a is a promising option for patients with not functioning or poorly functioning.
heart failure. The above-mentioned columnist Piet Borst (Textbox
A great many clinical trials involving gene therapy are at 11.1) wrote that after the Gelsinger drama there was
an advanced stage, but as far as we know there has only light at the end of the tunnel after all and referred to the
been one commercial treatment, and that was in China. group under Alain Fischer in Paris. This concerned the
In the September 2005 issue of the American journal treatment of a rare form of SCID, in which there was
Human Gene Therapy Chinese researchers describe a defective gene on the X-chromosome (SCID-X1).
the world’s first commercial gene therapy treatment As a result of this gene defect in SCID-X1 patients,
(Peng, 2005). It involved a recombinant adenovirus with precursor cells of the immune system do not develop
a human gene that suppresses certain types of head and into adult resistance cells. The patients therefore have

Chapter 11: Gene therapy: a panacea for genetic abnormalities? 197

 no resistance to infectious diseases. In people with et al., 2003). Initially the researchers thought that the
a properly functioning immune system, intruders are leukaemia occurred because the gene was inserted so
rendered harmless by specialised white blood cells unfortunately in the DNA as to accidentally activate an
(T cells). Precursor cells of white blood cells usually oncogene in the T-cells. An oncogene is a gene that can
develop under the influence of growth factors into convert cells into tumours. Four years later, however, a
effective white blood cells, but not so in SCID-X1 number of American and German researchers reported
patients. These children were therefore not resistant to in Nature of 27 April 2006 that the inserted gene itself
all kinds of germs that are easily fought off by healthy was the cause of the disease, and they concluded that
people. They used to have to live in a ‘bubble’ to far more time was required in gene therapy research to
protect them from infection. Now they are administered carry out experiments with laboratory animals, before
with the missing resistance cells by means of regular testing on humans.
bone marrow transplants. After the discovery of leukaemia, the research was
In 1999 the French researchers “harvested” precursor voluntarily discontinued and only continued with lower
cells from SCID babies with a bone marrow puncture. doses of corrective cells after the protocol had been
They infected these precursor cells with a ‘cripple’ virus, revised. In January 2005, however, traces of cell growth
in which the researchers had inserted the ‘corrective’ were discovered in a third child, this time as a result
gene. After three days of being exposed to this virus of another oncogene. This child and one of the first
in a test-tube, the precursor cells were put back in the two victims responded well to chemotherapy. Sadly,
body of the babies (Figure 11.2). After three months the other of the two did not, and died in October 2004.
of isolated nursing care, adult resistance cells were Edelstein et al. reported in 2007 that all the other patients
circulating in the babies’ blood. Since they now had a in the French study had to date benefited from the gene
properly functioning immune system, they were allowed therapy. On 31 January 2009 it was reported that eight
to go home. On 21 January 2001, Borst wrote: “In two of the ten had been able to live four years on average
of the babies the gene therapy took place more than a without medication, according to an international study
year ago, and they are still in great condition, according under the supervision of the gene therapy centre in
to Fischer. Fischer’s success is based on careful Milan.
preliminary research on cells in laboratory animals, so The latest news we have about the “bubble boy”
that he could find out precisely what the best method treatment is from 21 July 2010. Gene Emery writes for
was of carrying out gene therapy.” Two years later, Reuters that the 10-year study of nine boys born without
however, it seemed that it was still not enough, when the ability to ward off germs has found that gene therapy
the researchers published that two of the patients is an effective long-term treatment, but it comes at a
treated had contracted leukaemia (Hacein-Bey-Abina price: four of them developed leukaemia. He quotes the

198 Part 3: Health has limits

team leader of the Paris research group that reported therapy would be more successful if it were carried out
their conclusion in the New England Journal of Medicine: prenatally, on the embryo in the uterus. Embryonic cells
“All children except one, including the three survivors grow faster than adult cells and so take up “foreign”
of T-cell acute leukaemia, could live normally in a non- DNA more easily. Moreover, the immune system in
protected environment and cope with microorganisms the embryo is not yet fully developed, so there will
without harmful consequences while growing normally.” be less risk of the “foreign” substance being rejected.
Earlier that month, on 7 July 2010, Amy Dockser Aside from the fact that this form of gene therapy
Marcus wrote in The Wall Street Journal that begs all kinds of ethical questions (for example,
researchers have launched a new gene-therapy trial whether embryos should be manipulated), there are
for children with the rare disease known as “bubble boy also extra risks associated with it. If the treatment is
syndrome”. In the new study scientists plan to enrol 20 not performed well, the embryo and, potentially the
boys with SCID-X1 at five sites around the world. In mother, may develop an infection which could lead to a
this new trial researchers will take stem cells from a miscarriage. Another concern is that the inserted gene
patient’s own bone marrow, deliver a functioning gene ends up not only in the intended tissues but also in
into those cells in the lab and then infuse them back other tissues of the unborn child, for example, in the
into the patient. The researchers believe they have bone marrow, where it could cause damage.
stripped out the feature of the treatment that caused The most controversial issue in the discussions on gene
leukaemia. The parts of the vector thought to have therapy is the possible danger for future generations.
activated leukaemia-causing genes have been taken Such criticism is levelled not only at the treatment of
out. Study participants will be monitored for 15 years the unborn child, but also at gene therapy in general.
to rule out any cancer risk. There are clearly only risks for future generations if the
administered gene also gets into the reproductive cells,
11.4. GENE THERAPY IN THE UTERUS i.e. into the ova of the woman or the sperm of the man.
It does appear that DNA can go from one cell to another,
The above section seems to demonstrate that it is so in principle a little of the “foreign” DNA could reach
extremely difficult to insert a corrective gene into the the reproductive cells. However, it has been calculated
cells of patients in such a way that it functions well that the risk of this causing congenital disorders in the
and that the recipient of the gene gets better and offspring is extremely slight. What is clear is that much
stays healthy. According to Mels Sluyser (1999), a more research is needed to define good treatment
well-known cancer researcher as well as an artist of protocols. Only then will an informed decision be possible
some distinction , the question is whether the gene
75
on the question of whether or not it is acceptable to
conduct gene therapy on an unborn child.
75
www.mels-sluyser.com/Nederlands/overmels.html

Chapter 11: Gene therapy: a panacea for genetic abnormalities? 199

 The debate on preconceived genetic modification not feasible. There are two major theoretical problems
of reproductive cells is a whole different ball game, in changing our reproductive cells (aside from all the
particularly when it involves the insertion of desirable technical obstacles): directed change of a number of
characteristics (intellect, musicality, etc.) or the genes in the same ovum/sperm is impossible, nor is
‘erasing’ of undesirable characteristics (baldness, red there any theoretical solution for this problem in sight;
hair, etc.); that takes us into the arena of eugenics. altering the expression of complex characteristics
‘Made-to-measure children’ may well be a great topic like intellect requires a knowledge of genes and gene
for conversation today, but the reality, should society interactions that lies outside our imaginary powers.”
ever find that acceptable, is a long way off. Although technological advances often move at a
faster pace than we believe possible, we completely
PRECONCEIVED GENETIC MODIFICATION
agree with this statement, never mind the discussion
I WOULD LIKE TO ORDER as to whether or not it is even desirable.
ONE BABY TO GO ...
11.5. NOT EVERYTHING CAN BE TREATED (YET)
... MEDIUM SIZE,
LOW ON FAT WITH
EVERYTHING ON IT! Even if the genes responsible for all congenital
diseases were known, the use of gene therapy would
still be limited. This was obvious right from the early
days of gene therapy (Mariman, 1994). Some basic
knowledge of human genetics is invaluable in order
to be able to understand this, and you can find this
in Textbox 11.3. In principle gene therapy can cure
mainly the recessive genetic diseases. The dominant
or recessive nature of an inherited characteristic is
already determined at fertilisation. The sperm cell
delivers a complete set of genes to the ovum, which
The previously mentioned Emeritus Professor has a complete set of its own. So a fertilised ovum
Piet Borst was very clear about this in his column and the new cells resulting from this, excluding the
Eugenetische oprispingen (Eugenic burps) of 21 reproductive cells, contain a copy of each gene
October 2000. “I don’t think that anyone able to read this from the father and the mother. The symptoms of
column (eight years and older) will ever witness this, a recessive genetic disease only manifest if both
because all those carelessly proposed procedures are copies of a gene are defective. As long as one copy

200 Part 3: Health has limits

TEXTBOX 11.3. with the complementary half (or halves) of the genetic
Genetics in a nutshell. material of the diploid parent cell (2n). The haploid
cells thus formed are sex cells (gametes) which again
Chromosomes carry genes and regulate cell activity. produce cells via fertilisation (union of a male and female
They are made up of DNA with RNA and proteins. It gamete) with a complete set of chromosomes (2n).
is assumed that every chromosome has a double helix These cells have characteristics of both parent cells
which consists of two strands of complementary DNA whereby the relationship between them is determined
(see also Textbox 1.1). The number of chromosomes by a simple dominant/recessive relationship (Mendel’s
per cell nucleus determines the type. If there is one of Laws) between the alleles.
every chromosome, it is called a haploid cell. If there An allele is one of the two possible forms of a gene
are two of every chromosome (so-called homologous in a diploid cell. The alleles of a specific gene occupy
chromosomes), then it is a diploid cell. Humans are the same place (locus) on homologous chromosomes.
diploid and have two sets of 23 chromosomes, thus A gene can assume different forms (alleles) due to
46 in total, of which one pair is sex chromosomes. Cell mutations. A gene is homozygotic if the two loci have the
division is the process whereby a cell splits into two same alleles and heterozygotic if the alleles are different.
daughter cells. During this process the chromosomes If there are two different alleles present, one of them (the
duplicate – whereby the two strands of DNA form each dominant allele) suppresses the effect of the other (the
other’s matrix - and then split up during a process called recessive allele). The allele that determines the normal
mitosis, so that each daughter cell gets a package of form of the gene is usually dominant, while the mutated
chromosomes identical to the parent cell. Meiosis or cell is usually recessive. Most mutations are therefore
reductional division, in contrast, is the cell division expressed in the phenotype (outer appearance) if it is
process that leads to the formation of daughter cells homozygotic, i.e. if both alleles are mutated.

is healthy, the disease will not be present. So the of the defect in the dominant gene, other genetic
insertion of a healthy copy in the cells of a patient techniques, for example gene silencing, may offer a
with a recessive disease may lead to an improvement solution for some of the dominant inherited diseases.
or a cure. In dominant inherited diseases, however, In addition to recessive and dominant genetic disorders,
the symptoms always manifest when there is one where there are defective copies of a specific gene,
defective gene copy, even if there is a healthy second there is a big group of complex genetic diseases
copy of the gene in the cells. In this case, therefore, caused by a combination of different poorly functioning
the addition of another healthy copy will not lead to a proteins and adverse environmental influences. Well-
cure. Maybe in the future, depending on the nature known examples of this are rheumatic disorders and

Chapter 11: Gene therapy: a panacea for genetic abnormalities? 201

 the frequently occurring skin disease psoriasis. Several genes and experiments were already being carried out
genes at a time would have to be corrected in order for on patients with a congenital muscular disorder. The
these sorts of diseases to be effectively treated with first results, however, were disappointing. Injections of
gene therapy. This is a particularly challenging and, for myoblasts (the progenitors of muscle cells), to which
the time being, unfeasible exercise. an intact gene had been added outside the patient’s
body, did not strengthen the muscles of young people
11.6. GENE DOPING with Duchenne muscular dystrophy. The first genuinely
positive results of muscle-strengthening gene therapy
The term gene doping is relatively new. It first surfaced were seen in the late 1990s with mice and these led to
in the media towards the end of the 1990s, when discussions about gene doping. Journalists call such
the first publications about muscle-strengthening transgenic mice, half affectionately and half warily,
genetic modification of rodents began to appear. If Schwarzenegger mice. A more recent, spectacular
you “Google” the term gene doping now, you will get example of what gene doping can do to mice was
thousands of hits. published in the online journal PLoS in August 2007 by
Right from the first gene therapy experiments in Se-Jin Lee of the Johns Hopkins University in Baltimore.
the early 1990s, researchers were discussing the He describes a genetic procedure aimed at influencing
possibility of strengthening muscles by inserting new the formation of two proteins which regulate muscle

TEXTBOX 11.4. mutations in both versions of the gene. The father is

The German muscleman. not known to the doctors.

THE GERMAN MUSCLEMAN

In June 2004 the New England Journal of Medicine
reported the existence of a German muscleman (Van
Caulil, 2004). When he was seven months old he could
already stand and as a mere four-year old he could
hold dumb-bells weighing three kilos in outstretched
arms. So the 2020 Olympic Gold for weightlifting is
virtually a given for Germany. But this is not the result of
the notorious East German doping programme of that
time. It is nothing more than Mendelian inheritance.
His mother was a strong athlete, with one version
of a defective myostatin gene. Her precious son got

202 Part 3: Health has limits

growth, namely myostatin and follistatin. Myostatin with most other forms of doping. WADA had a broad
inhibits muscle growth, while follistatin deactivates definition for gene doping (Schjerling, 2004): “the non-
the effect of myostatin by binding to it. The aim of the therapeutic use of cells, genes, genetic elements, or of
procedure was to deactivate the myostatin gene and the modulation of gene expression, having the capacity
continually activate the follistatin gene. The resulting to improve athletic performance.” Meanwhile it has been
transgenic mice acquired four times as much muscle revised to: “The transfer of cells or genetic elements or
mass. This result was a lot better than expected and led the use of cells, genetic elements or pharmacological
to the suspicion that a few other unknown mechanisms agents to modulating expression of endogenous genes
play a role in muscle formation. Lee advises athletes having the capacity to enhance athletic performance,
not to experiment with this, but he hopes to be able to is prohibited.” Schjerling is a researcher at the Muscle
help people suffering from muscular dystrophy, AIDS Research Centre in Copenhagen, an institute that
or cancer. examines the question of how sportsmen and women
The tremendous impact on muscle growth of could raise their level to achieve medal status with
deactivating the myostatin gene was nothing new. The gene therapy. In November 2003 the Rathenau Institute
Belgian Blue cattle are blatant proof of how effective organised a symposium on the ‘makeable’ man. This
it can be. These animals are popular among cattle institute shows the impact of science and technology
breeders: they have more muscle mass, a stronger on our daily life and charts the dynamics of this impact
skeleton and less fat. All without any training - they by conducting independent research and debate.76
are lethargic beasts! The myostatin gene of these Schjerling said at this symposium: “I don’t expect to
cattle contains an error, which means that they can’t see any genetic doping at the 2004 Olympic Games
make myostatin. Moreover, the cows have traditional in Athens. … The risks at this stage are too great. …
breeding to thank for this phenomenon, not gene Research is not advanced enough.” During the Olympic
therapy or genetic modification. A myostatin deficiency Games in Athens there were frequent statements
can also turn a man into Samson (Textbox 11.4). in the media expressing the expectation that this
would be the last Games without gene doping. Hidde
The transgenic musclemen are not only arousing Haisma, a professor in Therapeutic Gene Modulation
interest in the meat industry (Textbox 11.5). The WADA, at the University of Groningen, also supported this
the world authority in anti-doping, is also eager to conclusion in 2004, having spent that year analysing
know more. As far as is known gene doping has not the possibilities and risks of gene doping at the request
yet been used in the sporting world. And yet it has of the Netherlands Centre for Doping Issues (NeCeDo).
been on WADA’s official black list since 1 January However, it now seems that the 2008 Olympic Games
2003. That’s the reverse way of doing things compared
76
www.rathenau.nl

Chapter 11: Gene therapy: a panacea for genetic abnormalities? 203

 TEXTBOX 11.5. and/or meat production in these so-called ‘knock
Hormone mafia becomes gene mafia. outs’. There is already a lot of practical experience
with animals who by nature have one myostatin gene
For years Willem Koert has been a scientific journalist that doesn’t work or works inefficiently. For example,
on the weekly paper at Wageningen University. In the it is well known that there are varieties with a poorly
issue that appeared on 14 October 2004 he wrote functioning myostatin gene that do not suffer any
an article on gene doping with the same headline as health problems. And yet breeding programmes are
this textbox. The subtitle of the article was as follows: thwarted by dystocia. This means that the calves
“Illegal fiddling with genetic modification untraceable.” become so big in the uterus that it is impossible to bring
The summary stated: “Doping detectives think that the them into the world naturally. A Caesarean section is
first gene technologically changed top athletes will be always required. Randomly intervening in the genome
making their appearance within the next few years. of animals in order to deactivate the myostatin gene
Will the hormone mafia follow this example and feed is therefore not a real option, never mind all kinds of
cattle using ‘gene doping’? The clandestine potential other possible complications. So we are in complete
of an experimental technology.” agreement with Koert. Consequently, to end with, his
At the moment the hormone mafia are still using old- central magnified, fairly crass statement: “If idiots start
fashioned preparations such as anabolic steroids experimenting with this technology in underground
and clenbuterol, but who will be able to stop them labs, I dread to think what will happen.” And so do we!
from using gene technology now that the possibility
GENE DOPING IS BIG BUSINESS
has come enticingly close? Researchers from John
Hopkins University have published prolifically about the
myostatin gene. Due to a defect in that gene “double- GENE DOPING

muscle cows”, such as the Belgian Blues, make none

or hardly any of the hormone protein myostatin and as a
result acquire extraordinarily big muscles. Using gene EPO -5,6% | STEROIDS -3,2% | GENE THERAPY
+ 10,1% | THG -1,2%

technology, this gene could in principle be deactivated

in healthy cows, but there remains considerable doubt
as to whether that alone will increase the muscle growth

in Beijing were also free of gene doping. Of course, we of urgency. WADA finances most of these research
can’t be absolutely certain because there are no tests projects. Françoise Lasne, who as we saw in Chapter
yet. But these are being developed with some degree 10 has been responsible for so much groundbreaking

204 Part 3: Health has limits

doping test research, has already shown that doping essentially entirely mapped (see Chapter 13). The
with the gene for EPO, injected into key muscles before biggest task is yet to be tackled, though, and that is
a sporting performance, can be traced. She conducted the identification of all genes, their functions and the
the experiment on macaques. However, the complexity way in which they operate. Only then will it be possible
of the matter leads to the unfortunate suspicion that this to point to the genes responsible for specific diseases.
is yet another case of lagging behind events. The technique itself will also have to be substantially
improved, in order to properly insert corrective genes
11.7. GENE THERAPY: NOT YET A PANACEA into the cell, so that they have a lasting desirable effect.
OR A REVOLUTION In short, neither panacea nor revolution just yet - but
hopefully, as Winston Churchill once said, it is the end
As mentioned earlier, more than 4,000 diseases and of the beginning!
abnormalities, ranging from SCID to cystic fibrosis, are That success in gene therapy is still a long way off
caused by a congenital defect in a single gene. Many is evidenced by the fact that there have still been no
other disorders, including cancer, heart defects, HIV recorded cases of gene doping. Sporting history is full
and senility, are to some degree the result of postnatal of examples of athletes suffering premature death,
damage to one or more genes. It is also clear that cardiac dilation and other ailments in return for a
gene therapy is still far from being a panacea for all higher chance of victory. A who-dares-wins approach!
diseases caused by a defect in one single gene. Yet, That was also the message from Mark Frankel of
in this chapter, we have also shown that there have the American Association for the Advancement of
been a few successes, albeit with eventual setbacks, Science (AAAS) at the gene doping symposium in
in the treatment of some serious genetic abnormalities. St. Petersburg in June 2008 (Köhler, 2008). There he
For the time being this will not be the case for many demonstrated that the huge financial stakes would
other genetic disorders because there is limited or no guarantee that gene doping will be used as soon as
knowledge about their genetic foundation. However, possible. As Frankel says, sport, medicine and science,
this situation is likely to change rapidly in future it’s all business. And right in the middle, there’s gene
decades, now that the human genome has been doping! We’re very much afraid he may be right.

Chapter 11: Gene therapy: a panacea for genetic abnormalities? 205

 11.8. SOURCES afweerreacties op gentherapie worden niet gemeld.
NRC.
Bainbridge, J., Smith, A., Barker, S., Robbie, S., Henderson, Köhler, W. (2008, 26 July). Epodope. NRC.
R., Balaggan, K., et al. (2008). Effect of gene therapy Mariman, E. (1994). Gentherapie gaat erfelijke ziektes te
on visual function in Leber’s congenital amaurosis. New lijf. Chemisch Magazine (March), 104-106.
England Journal of Medicine, 358(21), 2231-2239. Peng, Z. (2005). Current status of gendicine in China:
Blaese, R., Culver, K., Miller, A., Carter, C., Fleisher, T., recombinant human Ad-p53 agent for treatment of
Clerici, M., et al. (1995). T lymphocyte-directed gene cancers. Human Gene Therapy, 16(9), 1016-1027.
therapy for ADA-SCID: initial trial results after 4 Raper, S., Chirmule, N., Lee, F., Wivel, N., Bagg, A., Gao, G.,
years. Science, 270(5235), 475-480. et al. (2003). Fatal systemic inflammatory response
Cavazzana-Calvo, M., Hacein-Bey, S., Basile, G., Gross, F., syndrome in a ornithine transcarbamylase deficient
Yvon, E., Nusbaum, P., et al. (2000). Gene therapy patient following adenoviral gene transfer. Molecular
of human severe combined immunodeficiency Genetics and Metabolism, 80(1-2), 148-158.
(SCID)-X1 disease. Science, 288(5466), 669-672. Rosenberg, S., Aebersold, P., Cornetta, K., Kasid, A.,
Couzin, J., & Kaiser, J. (2005). Gene therapy - As Gelsinger Morgan, R., Moen, R., et al. (1990). Gene transfer
case ends, gene therapy suffers another blow. into humans--immunotherapy of patients with
Science, 307(5712), 1028-1028. advanced melanoma, using tumor-infiltrating
Edelstein, M. L., Abedi, M. R., & Wixon, J. (2007). Gene lymphocytes modified by retroviral gene transduction.
therapy clinical trials worldwide to 2007 - an update. New England Journal of Medicine, 323(9), 570-578.
Journal of Gene Medicine, 9(10), 833-842. Schjerling, P. (2004, 1 November). With good comes
Edelstein, M. L., Abedi, M. R., Wixon, J., & Edelstein, R. M. the bad: the reality of gene doping; the misuse of
(2004). Gene therapy clinical trials worldwide 1989- gene therapy by elite athletes is inevitable. Genetic
2004 - an overview. Journal of Gene Medicine, 6(6), Engineering News.
597-602. Sedlak, B. J. (2003, 15 May). Possibilities move forward for
Friedmann, T., & Roblin, R. (1972). Gene therapy for human gene therapy; gene activity therapeutics enter phase
genetic disease? Science, 175(4025), 949-955. II trials. Genetic Engineering News.
Hacein-Bey-Abina, S., von Kalle, C., Schmidt, M., Le Deist, Sluyser, M. (1999, 17 April). Gentherapie in de baarmoeder.
F., Wulffraat, N., McIntyre, E., et al. (2003). A serious Telegraaf.
adverse event after successful gene therapy for Van Caulil, G. (2004, 17 September). Transgene strijd –
X-linked severe combined immunodeficiency. New Gentherapie zal de weg volgen van menig ander
England Journal of Medicine, 348(3), 255-256. geneesmiddel: een toepassing als doping. Bionieuws,
Köhler, W. (2000, 29 January). Fatale haast; meeste pp. 8-9.

206 Part 3: Health has limits

12 XENOTRANSPLANTATION

“People are entitled to disagree with xenotransplantation, but then they should register as organ donors.”

The above statement was made by Guido Persijn, former Medical Director of the Eurotransplant Foundation,
an international organisation that coordinates organ donation and transplantation. Xenotransplantation is
the transplantation of organs, tissues or cells from one species of animal to another. This chapter will look at
transplantations between animal and humans. Xenotransplantation is one possible solution for the organ donor
shortage in the area of transplant medicine77. However, there is still a ban on this type of procedure because of
the lack of clarity about the sort of risks entailed. The natural rejection responses to cross-species components still
create insurmountable problems. The transfer of viral DNA with, as yet, unpredictable consequences is also another
matter that requires due attention. The various facets of this topic will be discussed in this chapter, as will the question
of whether or not xenotransplantation is ethically responsible. We’ll begin with the history of xenotransplantation,
which has its origins in a dark past.

XENOTRANSPLANTATION IS GREAT
STUFF FOR BACHELOR PARTIES
I THOUGHT A BUNNY SUIT
WOULD BE ENOUGH!

77
www.eurotransplant.org/?id=xeno

J. Tramper, Y. Zhu, Modern Biotechnology – Panacea or new Pandora’s box? 207

DOI 10.3920/978-90-8686-725-7_12, © Wageningen Academic Publishers 2011
 12.1. THE HISTORY OF al. (Deschamps, Roux, Sai, & Gouin, 2005). Because of
XENOTRANSPLANTATION: A SHOCKING PAST my personal interest in the subject of organ donation, the
article read as a real horror story, so incredibly thrilling,
Don’t take your organs to heaven; heaven knows we that I felt almost guilty for reading it ‘in the boss’s time’. It
need them here. became the most important source of information for this
chapter. The Dutch book was published in November
This quotation, whose origins are unknown to me (JT), 2009. Although we did quite a bit of revision, it remains
hangs on the wall in my sister’s bathroom. For more than also one of the main sources for this English version.
ten years, she has been hosting one of my brother’s Long before there was any insight into
kidneys (see Textbox 12.6). It was a perfect match! So xenotransplantations of whatever kind, there were stories
you’ll understand my particular interest in this topic. I in folklore about creatures that were half-man, half-beast
wrote the first draft of the Dutch version of this chapter in (chimeras). Pre-historic cave drawings seldom showed
2001 during a sabbatical in Lausanne. It lay untouched people, but in the Lascaux caves in France there is
until 2007 when in December of that year my co-author a drawing of a man with a bird’s head (circa 15,000
(YZ) updated it. On 28 October 2008 I began the final BC). The first description of what can be defined as
revision of the Dutch version by reading an article on the xenotransplantation comes from Indian mythology, in a
history of xenotransplantation, written by Deschamps et Sanskrit text from the 12th century BC (Textbox 12.1).

TEXTBOX 12.1.
The first xenotransplantation.

Shiva and Parvati are two Gods from Indian mythology.

According to the legend, their child Ganesha was born
while Shiva was out hunting. As in so many myths,
Ganesha was born a giant. When Shiva returned home
and saw his wife with this gigantic ‘stranger’, he beheaded
him. Parvati informed him that he had just killed his own
son, and threatened to destroy the universe if Ganesha
was not resurrected. Shiva, who wasn’t able to re-attach
Ganesha’s head, ordered his servants to bring him the
head of the first living creature they encountered. And so
Ganesha was given new life with the head of an elephant.
Source: Shutterstock

208 Part 3: Health has limits

Going back many centuries before Christ there were clearly show is that the idea of extending or enhancing
already descriptions of transplantation experiments life, by replacing failing or missing organs with organs
with people. For example, Susrata, an Indian from a human donor, was already around a long time
surgeon, was said to have used pieces of skin, about ago.
600 years before the calendar era, to replace noses
that had been cut off (often as a punishment). The The first xenotransplantations were carried out at
most notorious example of early transplantations is the beginning of the sixteenth century with cells and
probably ‘the miracle of the black leg’ from the 3rd tissues from bone, skin, and testicles, etc., the latter
century AD. In Rome two Syrian doctors, Cosmas and playing a particularly important role (Textbox 12.2).
Damian, amputated the gangrenous leg of a verger. Organs were only used much later on, because for a
They attached in its place the leg of a dead black long time there was no technique to keep the bleeding
man. All these early attempts were transplantations under control once the diseased organ had been cut
from human to human (allotransplantation); all were out, and no way to restore blood circulation after the
probably not very successful, even though the species transplant.
barrier had not been crossed. However, what they In 1668 the Dutchman Job van Meekeren reported a
successful xenotransplantation with bone, performed
SOMETHING WENT WRONG by a Russian who used a piece of dog skull to repair a
human skull. His claim that this had never been done
I’M SORRY, THERE WAS A SMALL MIX UP
before was later refuted. In 1501 the Iranian physician
WITH YOUR ALLOTRANSPLANTATION.
Muhammad Baha’ al-Dawla published medical notes in
YOU’RE SORRY? The Quintessence of Experience. In it he described the
surgical removal of a piece of skull that was infected
and replaced by a piece of dog bone, the brain being
protected by a piece of cucumber. He also reported
that in Herat, Afghanistan, the Indian surgeon Ala-ul-
Din had used fresh canine skin to replace all the skin
on the head of a patient suffering from eczema. These
early experiences were followed by many other similar
primitive experiments. The first person to describe it
as a transplantation was the Scottish surgeon John
Hunter in 1778, when he wrote of a transplantation of a
human tooth to the comb of a cockerel.

Chapter 12: Xenotransplantation 209

 Xenotransfusions are also centuries old. The parliament, quickly followed by the English parliament
transfusion of lamb’s blood to a 15-year old boy on 15 and then by the Pope. Despite this ban, documents
June 1667 is the first documented account of such a describing xenotransfusions continued to appear,
procedure. This was performed by the French doctor even until quite recently. In 2000 the Indian Dhani
Jean-Baptiste Denis, King Louis XIV’s physician, and Ram Baruah administered more than quarter of a litre
the surgeon Paul Emmerez in Paris. Less than half of pig’s blood to the 22-year-old Hussan Ali, who was
a year later, on 23 November, Richard Lower carried suffering from severe anaemia. Ali was discharged
out the same experiment in London on the 22-year- from hospital four weeks later. Tests confirmed that he
old Arthur Coga, also successfully. Several other had non-human blood cells in his bloodstream. A few
transplantations followed, but with less success, and years ago, fresh blood shortages also led to calls to
on 10 January 1670 they were banned by the French reconsider xenotransfusions. Artificial oxygen carriers

TEXTBOX 12.2. first procedure on a man, using the testicles of a

Human rejuvenation transplants. chimpanzee; slithers of testicle were inserted into the
scrotum. Three years later 43 men had undergone this
In 1889 the French-American doctor and physiologist operation and in 1930 that figure rose to 500. Women
Charles-Edouard Brown-Séquard injected himself received an ovary from female apes for the treatment
subcutaneously with an aqueous extract of crushed of menopause. Yet more shocking is the fact that he
testicles from dogs and Guinea pigs. These injections inserted a human ovary into a female chimpanzee
were intended to restore his physical strength and (Nora) and inseminated her with human sperm; so
capacities that were diminishing due to the ageing much for ethics!? The insemination was unsuccessful,
process. In so doing the 72-year-old Brown-Séquard but Nora did get the leading role in the 1929 novel
invented opotherapy, a treatment using bodily fluids “Nora the she-monkey becomes a woman”.
and a forerunner to endocrinology. Since then, During his career Voronoff was concerned about
numerous medicines based on crushed animal the adequate supply of donors (monkeys), and
organs have come onto the market; extract of thyroid considered setting up monkey farms in French
and pancreas are still available. Serge Voronoff Guyana in which to breed monkeys for export. Vilified
turned this therapy into a surgical procedure. Born in by the scientific community and the public, Voronoff
Russia in 1866, Voronoff acquired French citizenship stopped performing after 2000 xenotransplantations.
in 1895. He wanted to rejuvenate men by means of He died in 1951. It should, however, be noted that he
xenotransplantation with the testicles of chimpanzees was the first person to draw attention to the problem
and baboons. On 12 June 1920 he carried out the of donor shortages.

210 Part 3: Health has limits

for blood transfusions have also been attracting received a kidney from a chimpanzee. When she died
attention in research circles in recent years. nine months later, an autopsy revealed the only cause
The most important criterion for successful organ of death to be an acute imbalance of electrolytes.
transplantation is the restoration of the vascular Nine months surviving without rejection of the liver
tissue of the organ in question by the stitching delivered proof that xenotransplantation was possible
together of the arteries. More than a century ago, in principle. Yet the results remain unimpressive
the Frenchman Mathieu Jaboulay and his student and this nine-month survival is still a record. In
Alexis Carrel pioneered this technique, using mainly 1976 the Swiss Jean-François Borel discovered
kidneys. Kidneys were preferred because it was easy cyclosporin A, a drug which was also expected to
to prove the success of the operation: you just had to be capable of suppressing rejection reactions in
wait until the patient urinated! xenotransplantations.
In January 1906, they attached the kidney of a pig, On 26 October 1984 the American Leonard Bailey
slaughtered three hours before, to the inside of carried out the operation that would become the most
a woman’s elbow. In the next day and a half they famous xenotransplantation in history. It concerned
collected one and a half litres of urine, but on the third the premature twelve-day old Baby Fae who had a
day they were forced to remove the kidney because heart defect. She was given a baboon heart. The
of thrombotic symptoms. Three months later they conditions seemed optimal, including the fact that
repeated the same operation with the same result cyclosporin A was now available. In addition, of the
in a different woman, this time with a goat’s kidney. six available baboons, the one chosen gave the
These transplantations are often regarded as the first weakest response with white blood cells and would
real xenotransplantation experiments, even as the probably cause the least rejection reactions as a
first organ transplantations. In 1909, 1913 and 1923 result. After eleven stable days, however, the first
other researchers carried out a xenotransplantation rejection symptoms were observed and 20 days after
with a kidney from a macaque, a Japanese monkey the xenotransplantation Baby Fae died. Much of the
and a lamb, respectively, but the results weren’t hope vested in xenotransplantation died with her and
much better. There then followed a period of 40 years a moratorium followed de facto. The new anti-rejection
without further attempts. drug, Tacrolimus, which was brought onto the market
The failure of these first experiments was the direct in 1992, did little to change this situation. The arrival
result of a lack of means to prevent rejection by the of transgenic pigs the same year, however, did seem
immune system. When these means came in the to herald change and a new era.
early 1960s, the interest in (xeno)transplantations
was reawakened. In 1964 a 23-year-old woman

Chapter 12: Xenotransplantation 211

 12.2. THE TRANSGENIC ‘SPARE-PART PIG’ functions. But, after several kidney transplants in the
1960s from chimpanzee to human, the entire medical
“The creatures outside looked from pig to man, community has agreed that ape organs are not a real
and from man to pig, option. This has to do with anatomical differences,
and from pig to man again; animal ethics and practical problems with breeding, but
but already it was impossible to say which was which.” probably more to do with the growing understanding
that non-human primates are a source of viruses that
George Orwell: Animal Farm; 1945 can be or can become very dangerous for humans.

Animal Farm Pigs are now the animal of choice. The anatomy of
their internal organs shows major similarities to those
of humans. They are also available in abundance and
have a relatively short reproduction cycle with big
litters. Years of experience also show that pigs are
relatively easy to breed in germ-free conditions. But
from an evolutionary point of view, pigs are a lot further
away from humans. This results in all kinds of acute and
chronic rejection symptoms. Genetic modification of
the pig and immune suppression in the organ recipient
George Orwell should overcome that. But even in the absence of
these obstacles, there still remains the question of
Source: Identim
whether the physiology of the pig is similar enough
Anyone looking at animals to solve the shortage of to that of humans. It is, as yet, a largely unexplored
transplant organs, should actually focus on those area of xenotransplantation: the comparison of
species that are evolutionarily most closely related, physiological and biochemical characteristics of man
e.g. primates (mammal group that includes monkeys, and pig; the subtle differences in hormone regulation,
apes and human beings). Apes are therefore the mineral concentrations and blood pressure.
recommended organ donors, especially since it now The size of hearts, kidneys, lungs, liver and blood
appears that there is a less than 1% difference in vessels in pigs are a good match with those of
genetic make-up between chimpanzee and man. That humans, making pigs ideal organ donors. But without
would seem to imply that there would be few rejection special precautionary measures, xenotransplantation
symptoms and only slight differences in the organ is guaranteed to fail. The rejection reaction is so

212 Part 3: Health has limits

strong, that the animal organ can be irreparably already have an antibody. This may be because we are
damaged within a few minutes. This sort of hyperacute confronted early on in life with bacteria which have the
organ rejection also occurs if a patient gets a human same sugar chain on the cell surface (Prather, 2007).

O A B AB

Pig sugar side chain

Galactose N-acetylgalactosamine L-fucose

1 Rejection

Anti body

2
No
rejection

3
No
rejection

Figure 12.1. The fight of the blood groups, adapted from Van Zundert (1998).

organ from a donor with a different blood group. Blood So our blood reacts directly to the pig organ, resulting
groups are determined by the nature of the sugar chain in hyperacute rejection (Reaction 1).
of the red blood cells (Figure 12.1). However, these Up until the turn of the century, getting around this
blood group determinants are not only found in red immune reaction by modifying the blood group
blood cells; they are also present in most organs, cells determinants in the pig was more of a theoretical
and tissues in the body. In pig organs, as well as in the than a practical or feasible solution. That’s why, at
pig cells that line the inside of the pig’s arteries, there the beginning of the 1990s, Imutran, a subsidiary of
are blood group determinants, which are recognised the pharmaceutical giant Novartis, tried a different
by our immune system as foreign, and for which we approach. Researchers there succeeded in genetically

Chapter 12: Xenotransplantation 213

 TEXTBOX 12.3. to the body’s own proteins or cells. This phenomenon
The immune system – some basic facts. is called self-tolerance. One reason for the strong
immune responses induced by viruses, for example,
The immune system is a remarkably adaptable is their particulate and repetitive structure. Viruses
defence system that has evolved to protect the body have small genomes and only a few different proteins
from invading microorganisms such as bacteria and available to build up a viral particle. The proteins
viruses. It vigorously fights such pathogens with an are therefore arranged in a highly repetitive and
armada of specialised cells and molecules, including highly organised format. There exists no comparable
the antibody-producing B cells and the T cells. B structure within the human body where a body cell
cells and T cells are the main effectors of the immune always has a few hundred to a few thousand different
system to establish effective and long-lasting immune proteins on its surface. During human evolution, the
responses and they are also the cell types that need immune system appears to have learned to recognise
to be targeted for successful vaccination strategies. such highly repetitive structures as foreign and
While B and T cells respond efficiently to foreign potentially harmful, and it reacts accordingly with a
invading viruses or bacteria, they usually don’t react potent immune response.

modifying pigs so that they produced a human protein immune processes, in order to better understand them
that could block the immune reaction between pigs’ so that more effective means to combat them can be
organs and human blood (Reaction 3). Testing with developed. In 2007 recent progress, the state of affairs
apes showed that the hyperacute rejection did indeed and future possibilities were looked at by Yang and
fail to occur. And yet, the pig organs failed in the long Sykes (Yang & Sykes, 2007a, 2007b) in two leading
run. Preventing a hyperacute immune reaction was journals. These data are not easy to summarise, but
clearly not the only challenge. At a later stage the body suffice it to say, there is still a long way to go. On the
throws in a whole army of antibodies and different website78 of Cytos Biotechnology we found a short
processes into the fight to take out the ‘intruders’. and simple summary of the immune system (Textbox
Even when there is a “perfect match” between human 12.3). Cytos Biotechnology is a Swiss company that
donor and recipient (Textbox 12.6), a transplant only is developing and commercialising a novel class of
succeeds by at least temporarily suppressing the medicines – called Immunodrugs™. Immunodrugs™
immune system with medication. However, it is still not are therapeutic vaccines intended for use in the
known whether this medication is effective enough in treatment and prevention of common chronic diseases
the case of a xenotransplantation. Which is why there which afflict millions of people worldwide.
is lots of research taking place into these complicated
78
www.cytos.com

214 Part 3: Health has limits

So to start with, a successful xenotransplantation 1990s on the genetic modification of pigs with a
involves preventing hyperacute rejection (Reaction 1). human gene. The gene in question codes for the
The most obvious solution is to give the pig the sugar protein hDAF (human decay-accelerating factor), one
chain that corresponds to the determinant of our blood of the proteins that inhibits the so-called complement
group ‘O’, because there are no antibodies for this sugar activation in humans (Siegert, Van Es, & Daha, 1996).
chain (Reaction 2). This means that only the extremity Complement activation is the result of the cascade-like
of the sugar chain, the galactose α-1,3-galactose interaction of plasma proteins and membrane-bound
epitope (an epitope is the location on the antigen which proteins. There is a total of approximately 30 proteins
can be specifically recognised by antibodies that will in the complement system. The complement system
bind to it) in the blood group determinant of the pig, will traces intruders in the bloodstream and destroys
have to be changed: a fucose in place of a galactose. them by drilling through their cell membranes. In a
The enzyme that links the latter sugar unit (galactose), transplanted organ they bind to the endothelial cells,
α-1,3-galactosyltransferase or α-1,3-GalT for short, the ‘inner lining’ of the blood vessels. These cells are
must be deactivated and an extra enzyme must be immediately destroyed. As a result coagulation factors
‘incorporated’ to ensure that the fucose attaches to the are released from the underlying cell layers, causing
right location. Deactivating a gene entirely is always the blood vessels to be blocked within minutes. Pierson
a complicated process, which until recently only had III et al. (2009) have nicely pictured and described
a reasonable chance of success in less complex this process in their invited review article “Current
animals such as mice, resulting in so-called “knock- status of xenotransplantation and prospects for
out” mice. Since 1992, however, researchers have clinical application” (Textbox 12.4). The complement
been working hard on the genetic modification of system has a number of safety markers where the
pigs for the purposes of xenotransplantation. In their induced reaction can be stopped (this is necessary if a
review ‘Xenotransplantation: The next generation complement factor binds to the body’s own structure);
of engineered animals’ d’Apice and Cowan (2009) hDAF marks one of these points. This also applies to
address the questions: What to remove? What to proteins coded as CD59 and MCP, as well as hDAF
add? And how to do it? In their final section “Horses membrane-bound proteins79.
for courses?” they end with: “Sounds familiar … so
maybe a hurdler can run on the flat? Our approach is On 23 December 1992 the first hDAF transgenic pig
to try to build a multitalented pig and put him over a few was born; it was named Astrid. Three years later the
different courses.” American company Nextran produced transgenic
As mentioned previously, the English company pigs that expressed two of this type of protein, hDAF
Imutran (Cambridge) began research in the early
79
www.ntvg.nl/node/290588/print

Chapter 12: Xenotransplantation 215

 TEXTBOX 12.4. thrombin. Thrombin amplifies the clotting cascade by
Dysregulated coagulation in pig-to-primate (a) activating XIa (not shown), (b) activating platelets,
xenotransplantation. (c) cleaving fibrinogen into fibrin monomers that form
the primary clot matrix, and (d) activating factor XIIIa
Coagulation is occurring continuously within the (not shown), which cross-links fibrin monomers into
bloodstream, but is normally restrained by a network an insoluble clot. TFPI and thrombomodulin normally
of inhibitory pathways involving endothelial proteins inhibit coagulation on healthy endothelium, while
such as thrombomodulin and tissue factor pathway soluble antithrombins inhibit thrombin by forming a
inhibitor (TFPI) (Panel A). Increased coagulation complex with its active site.
is normally initiated when endothelium retracts or Porcine EC activation – whether by xenoantibodies,
becomes ‘activated’ by injury, in part because von complement, or other factors – results in loss of
Willebrand factor (vWF) is expressed and tissue factor natural anticoagulant proteins (TFPI, thrombomodulin)
(TF) is liberated into the circulation. The coagulation and acquisition of a procoagulant phenotype (Panel
cascade then becomes amplified by the factors shown B). In addition functional incompatibilities in the
(VIIa/TF complex, IXa, and Xa) which in turn activate coagulation system between pigs and humans cause

Human endothelium exposed to human blood

Exposed subendothelium

Thrombomodulin
‘Contact factors’ Anti-
vWF XII APC protein S Thrombins Fibrinogen
Thrombin
Factor XIa XIa + VIIIa Va + Xa
Platelets Exposed vWF Prothrombin Fibrin
TF + VIIa
active Tissue Factor
TFPI
Injured endothelium Normal clot formation

Porcine endothelium exposed to human blood

Exposed subendothelium

Thrombomodulin
‘Contact factors’ Anti-
vWF XII APC protein S Thrombins Fibrinogen
Thrombin
Factor XIa XIa + VIIIa Va + Xa
Platelets Exposed vWF Fibrin
Prothrombin
TF + VIIa
active Tissue Factor
TFPI
Injured endothelium Thrombosis

Figure 12.2. Dysregulated coagulation in pig-to-primate xenotransplantation, reproduced with permission (Pierson III et al., 2009).

216 Part 3: Health has limits

both inappropriate or accelerated thrombin formation inhibition. The relative intensity of clot formation, the
and inefficient restraint of clot activation. Our current net product of coagulation pathway enzyme effects,
hypothesis is that xenografts succumb to an otherwise is symbolised by arrow weight at the thrombin and
insignificant humoral or cellular immune response fibrin steps. Pathways where pig endothelial proteins
which amplifies endothelial injury and intravascular inefficiently dampen coagulation are indicated with
thrombosis, and becomes manifest as thrombotic hatchmarked red lines in Panel B. For simplicity, only
microangiopathy. The blue arrows designate cascade the activated clotting factor intermediaries and key
amplification steps, while the red lines identify loci of points at which regulation occurs are shown.

and CD59, with an even greater chance of stopping then there have been many more promising
rejection. The Texan Robert Pennington owes his life examples. For instance, a group at the University
to one of these pigs. In autumn 1997 the liver of this of Missouri-Columbia joined forces with Immerge
man, who was 17 at the time, suddenly failed. There Biotherapeutics to make an α-1,3-GalT knock-out of
was no donor available at the time. Dr. Marlon Levy, the Imutran pigs with an hDAF gene (Prather, 2007).
a transplant surgeon at Baylor University Medical This transgenic pig model was disseminated by the
Center in Dallas, offered to pump his blood outside ‘National Swine Resource and Research Center’81.
his body through a transgenic pig’s liver until a donor Further modifications are still necessary before pigs’
liver became available. One of the Nextran transgenic organs can be successfully transplanted to humans.
pigs, a sow later named Sweetie Pie by Robert, was The review of Klymiuk et al. (Klymiuk, Aigner, Brem,
transported to Dallas and slaughtered; its liver was & Wolf, 2010) provides an overview of the transgenic
connected outside Robert’s body to his bloodstream. approaches that have been used so far to generate
For nearly seven hours spread over three days until donor pigs for xenotransplantation, as well as their
a donor liver was found, the pig’s liver detoxified biological effects in in vitro tests and in preclinical
Robert’s blood thereby saving his life . 80
transplantation studies. As a future challenge they
The next major step forward was made in 2001, see the combination of the most important and
when two different groups announced that they had efficient genetic modifications in multi-transgenic
created transgenic α-1,3-GalT knock-out pigs and pigs for clinical xenotransplantation. Aigner et al.
then cloned them. A year later, on 25 July 2002, the (Aigner, Klymiuk, & Wolf, 2010) review the selection
first four double knock-out pig clones (with both gene of promoter sequences for reliable transgene
copies deactivated) were born at PPL Therapeutics, expression for this purpose.
the company that created Dolly the Sheep. Since Xenotransplantation is still a very experimental

80
www.pbs.org/wgbh/pages/frontline/shows/organfarm 81
www.nsrrc.missouri.edu

Chapter 12: Xenotransplantation 217

 procedure and no creature has yet survived a 12.3. PANDEMIC RISKS
xenotransplanted pig organ for any length of time,
not even a monkey. However, developments in The last few decades have seen the spread of new
recent years have been such that, particularly with infectious diseases such as Ebola, HIV, Creutzfeldt-
transgenic pigs, pre-clinical studies with “pig-to-other- Jakob, SARS and Mexican flu. These were probably
animal” xenotransplantations are running in various animal diseases by origin, which have now become
countries to further investigate the feasibility, and infectious for humans. This has raised great fears that
‘pig-to-human’ xenotransplantations are starting to xenotransplantation would exacerbate such mutations.
appear on the agenda again. Xenotransplantation of Xenotransplantation is therefore regarded as a serious
the insulin-producing islets of Langerhans from the risk for public health, because it brings with it the
pancreas is in particular expected to proceed quickly risk of transferring swine pathogens, in particular
from pre-clinical to clinical phase (Schuurman, viruses that are not endemic to humans (Louz,
2008). One reason for this is that less than five Bergmans, Loos, & Hoeben, 2008). If patients receive
percent of the islets of Langerhans in pigs have the immune suppressants and still have no immunity,
Gal epitope, meaning that the risk of hyperacute xenotransplantation can, in the worst-case scenario,
rejection is smaller than in other pig organs (Prather, lead to a global pandemic with a new life-threatening
2007). Consequently, no α-1,3-GalT knock-out pigs virus. Many exogenous viruses can be eliminated by
are expected to be needed, in contrast to other pig pathogen-free breeding, by selection and vaccination
organs. Rajotte (2008) states that, according to clinical of the donor animals and by adequate screening of the
research, the preparation and transplantation of the organs for xenotransplantation. However, O’Connell
islets must be SAS - Safe, Affordable and Simple: (2008) concludes that suitable facilities for looking
(1) safe with regard to the transfer of pathogens (see after donor pigs still need to be designed. These will
following section), (2) affordable within our health- require lots of money for investment and maintenance
care parameters, and (3) simple and reproducible and it will be a considerable time before donor pigs
production of transplantable islets with a minimum of from a suitable facility are made viable for clinical use.
regulatory control. In short, there’s still a long way to go just with regard to
Transgenic pig clones clearly signal the first breach facilities and protocols.
of the rejection barriers. Yet there are still no clinical The biggest concern is about the pig endogenous
trials involving humans. There is still too little data retrovirus (PERV), of which there are several copies
available on the extent of the risk of transferring in the pig genome. This concern goes back to 1997
potentially very infectious viral DNA. Concerns about when it was shown that PERV could infect human cells
this are huge. if they were grown in a test-tube. Further indications

218 Part 3: Health has limits

were demonstrated in 2000 by means of experiments And despite all the ‘moped drivers without helmets’
with mouse models. PERV was transmitted when islets and major donor events, it doesn’t look as though this
of Langerhans were transplanted from a pig’s pancreas situation will change any time soon. In Europe in 2005
to an NOD/SCID (non-obese diabetic/severe combined only 3,540 kidney transplants were carried out, and the
immune-deficient) mouse; NOD/SCID mice are mouse average waiting time was three years. The picture was
models that are diabetic and have a defective immune no different in 2008. At that moment there were about
system. This proves that animal viruses can be 75,000 and 11,300 patients on the waiting list for kidneys
potentially transferred during xenotransplantation to in the US and Europe respectively (Sprangers, Waer, &
humans. Since then this has been a subject of concern Billiau, 2008). There have been no drastic changes since
and discussion, not only among health authorities. then and from what we know now, xenotransplantation is
It has given rise to a precautionary approach, strict unlikely to change this scene for the time being.
regulation and even a moratorium in many countries The transplantation of animal organs to humans is
on clinical trials, at least with humans82. It is crucial that ethically acceptable, according to a Dutch committee of
we remain aware of the fact that xenotransplantation the Health Council in its published opinion in January
combines possible advantages for the individual 1998 to the Minister of Public Health. But the committee
patient along with the risk of serious, large-scale, new also concluded that, before surgeons can routinely
infectious diseases (pandemics). Basically, PERV proceed to xenotransplants, problems with rejection
requires and is undergoing a thorough risk assessment and infection must first be resolved. The committee’s
at this time. opinion was largely consistent with those that had
appeared earlier in the United Kingdom and the US.
12.4. SOCIAL AND ETHICAL ASPECTS The British Nuffield Council of Bioethics stated that
the breeding of pigs was the most acceptable solution
In 1998 about two and a half million Dutch people for xenotransplantation. Breeding monkeys for this
registered their organs for transplant after their death. purpose was deemed unacceptable, mainly because of
However, this generous gesture is a drop in the ocean. the greater risk of infection. Monkeys are more closely
On the global level the supply of donor hearts, kidneys, related to humans than pigs. Pathogens in monkeys can
livers and lungs has for years met only a fraction of the more easily adapt when they enter a human body, than
demand. In 2002, the number of people across the world bacteria and viruses from pigs. And yet there is still a risk
registered as waiting for an organ was over 250,000. of infection from pigs’ organs, as we saw earlier in this
Less than a third of those waiting received a transplant . 83
chapter.

82
www.fda.gov/BiologicsBloodVaccines/Xenotransplantation/default.htm
83
www.eurotransplant.org/?id=xeno

Chapter 12: Xenotransplantation 219

 In an interview in 2000, Maaike Werner, the press Faced with the same question, the virologist Ab
officer for the Dutch Society for the Protection of Osterhaus (Erasmus University Rotterdam) said: “I
Animals until 2005, gave the following answer to the can’t give a yes/no answer to that question. In terms
question of whether xenotransplantation is ethically of animal welfare, I think xenotransplantation should
responsible or not: “The Society for the Protection of proceed under the right conditions. We breed pigs for
Animals doesn’t think xenotransplantation is ethically meat already. But if I look at the risk of infection in the
acceptable. We are worried about the emergence of future, I’m not sure about it. It may be that we create
a new bio-industry for organs. Just because the meat new viruses using this technique; then I predict a
industry is accepted, doesn’t mean that the organ doom scenario like AIDS. Xenotransplantation should
industry should automatically be allowed. The meat only proceed very gradually under strict conditions.
industry has to meet certain criteria on animal welfare, For example, you can start transplanting parts of
the organ industry is very different. Just imagine a organs and test them for viral infections. Either way,
genetically modified pig living in a sterile room without developments in this area cannot be stopped. So we’d
daylight or straw before it has its throat slit. And many better adopt the right approach and make sure that
laboratory animals have already gone the same way. A there’s a plan in place should anything go wrong.”
pig’s heart has a smaller capacity than a human heart, In an interview in the same year, Jan IJzermans, a
so they want to make a donor pig do conditioning transplant surgeon at the Erasmus University Medical
exercises. Don’t you think that’s absurd?” Centre in Rotterdam, summarised the facts: “The
problem is that the discussion is still vague, because
EXERCISES FOR PIGS...
ONE OF THE ABSURD ASPECTS there’s no real idea of how great the dangers are. We
OF XENOTRANSPLANTATION? have to weigh up the interests of the individual patient
who could be saved with a donor organ, and the risk of
a new epidemic. In contrast to many other controversial
medical procedures, however, xenotransplantation
cannot be dealt with simply as a personal choice, if
the possibilities are there. Anyone with a pig’s heart
or kidney may constitute a risk for the entire health of
the public.”
At the turn of the last century, the Dutch Foundation for
the Consumer and Biotechnology used a subsidy from
the Ministry of Public Health, Welfare and Sport (VWS)
to organise a public debate on xenotransplantation.

220 Part 3: Health has limits

The aim of the debate was three-fold: to give the unnaturalness of the procedure, the crossing of human
public information, let them form an opinion on and animal, and possible adverse consequences for
the basis of this information, and then assess the quality of life, were also important considerations.
opinion. On 10 November 2000 the bioethicist Egbert Compared with other forms of organ replacement
Schroten (University of Utrecht) and the then VWS (existing and experimental), xenotransplantation was
minister Mrs Els Borst, opened this debate called found to be the least acceptable.
Should xenotransplantation be allowed? It followed Many believed that xenotransplantation should only
an information campaign on xenotransplantation that be allowed if there were no other ways of solving the
began in December 1999. During that time the press donor shortage problem. The risk of infection was
also devoted a lot of attention to this subject. In the first the most important concern, but for the majority of
months of 2001 citizens were able to go and discuss people this was not a defining enough reason to reject
xenotransplantation in various locations around the xenotransplantation. In any case, people didn’t expect
country. In addition, the science theatre Pandemonia xenotransplantation to be used as long as there were
staged the play “Dierbaar Leven” (loosely translated still uncertainties about the risks. Opponents objected
this means ‘life is precious’, but there is a double to the consequences for animals with regard to
meaning in the Dutch since “dier” means animal) animal welfare, genetic modification and the fact that
about xenotransplantation. The idea behind this was xenotransplantation is a new use of animals. Others
to get young people interested in the topic. There was found these consequences problematic too, but for
a website (no longer available) allowing for a debate them the purpose of the use (saving human lives) was
on the subject. The public debate was concluded at more important. Nor did the latter group see any basic
the end of April 2001 with a final meeting. In mid- difference between this and using animals for other
2001 the final report was published by the foundation. purposes, such as eating them.
The summary and conclusions are available online , 84
Young people were much more concerned about
unfortunately only in Dutch, but the gist of these is as surviving than adults. While adults were more inclined
follows. About half the people who interactively took to accept the end of their lives, young people were
part in the debate had no particular opinion for or more willing to calculate in certain downsides of
against xenotransplantation. Proponents point mainly xenotransplantation. In conclusion, we would like
to the possibility of saving human lives and solving the to mention that people also indicated the desire to
problem of donor shortages. Opponents place more be able to vote in the future as to whether or not
emphasis on the “makeability” of the body - the extent xenotransplantation goes ahead. Crucial in this regard
to which we are ready to interfere with the body. The was clarity on the criteria used by the government and
doctors when deciding on who gets which treatment.
84
www.weten.nl/webzine/nummer4_2001/pdf/bruikbaar.pdf

Chapter 12: Xenotransplantation 221

 That, then, was the final report on the public debate. social and regulatory framework for clinical
A Dutch website that is constantly concerned with this xenotransplantation. She does so on the basis of an
theme is that of the working group on transplantation analysis of recent literature on regulatory questions
questions, which critically analyses all the medical, concerning xenotransplantation. She concludes
moral and risk aspects of transplantation85. There that the global scale on which the research is
is also a lot of relevant information to be found currently taking place, requires that some aspects
abroad, for example, from Canadian supporters and86
of xenotransplantation should be reconsidered.
American and British opponents.
87 88
Inadequacies and weaknesses in national legislation
Little has happened in the Netherlands since the big can, in her opinion, not only have undesirable
debate. Xenotransplantation has received very little local effects, but international implications as well.
attention and not much has changed as regards Conversely, the lack of international implementation
the thorny points. The global picture is very similar. of rules or “loose” interpretation of standards has a
What we do see is that there are still many scientific negative impact on groups and populations who are
publications and review articles appearing. George already disadvantaged, and may result in potential
(2006) concludes in his article that xenotransplantation risks worldwide. Although specific subjects such as
will probably remain controversial because of the animal welfare and rights continue to be discussed,
complex nature of the medical, ethical and legal the most important aspect of the regulatory questions
questions. If the scientific problems were to be solved, concerning xenotransplantation is increasingly
the decision to proceed with the clinical application shifting to the multi-faceted aspects of locality and
of this technique would depend, in his view, on a globality, where space, as well as formal legislation, is
collective decision based on ethical, regulatory and created for non-governmental networks as potentially
legal frameworks arising from a consensus. What is flexible and normative instruments. In short, if we the
clear is that there is still much opposition. Googling authors understand it correctly, there’s still a long way
Frankenstein in combination with xenotransplantation to go.
yields thousands of hits, generally not very friendly So far, the religious angle has received little attention
with respect to this topic. in the press and scientific literature. We have only
In recent reviews, Professor Mariachiara Tallacchini been able to find one recent article on this (Bruzzone,
(Tallacchini, 2008; Tallacchini & Beloucif, 2009) 2008), which basically concludes that no religion has
defines what she sees as a suitable ethical, an official ban on xenotransplantation!

85
www.stelling.nl/xeno
86
www.islet.org
87
www.crt-online.org
88
www.uncaged.co.uk

222 Part 3: Health has limits

 12.5. IN CONCLUSION surrounding xenotransplantation if you completely ban
research in this area? And even with research, there
Xenotransplantation clearly stirs society’s conscience. is still the question of how science can acquire real
This social anxiety is mainly to do with the risk that insight into the risks without exposure to those risks.
transplanting an animal organ to a human body may It’s not always possible to rationalise. It is fortunate that
wake a sleeping virus. The pig genome may contain the decision rests not only on scientists’ shoulders, but
viruses that are harmless to the pig. All they do is sit in also on those of the whole of society.
the DNA and replicate. Nothing more. It could be, with Companies are also wrestling with this problem.
the emphasis on ‘could’, because nobody knows for Geron Bio-Med in Roslin, Scotland, a company trying
sure, that these viruses mutate in a human environment in various ways to apply the cloning technique used
into a variant that is harmful to humans. It would be to “make” Dolly the Sheep and to make money out
unlucky indeed to have created with our own hands a of it, reported in mid-2000 that it was getting rid of its
brand-new viral infection to rival, for example, HIV. potential donor pigs. The company was planning to
focus on embryonic stem cells that can be converted
THERE ALSO ARE SOME UNEXPECTED COMPLAINTS in the laboratory into specialised human cells and
AGAINST XENOTRANSPLANTATION

?
tissues for all sorts of medical applications (Chapter
14). The hope is that whole organs can be made in
CAN I GET A GOAT HEART this way. Human, not animal, organs - from our own
TRANSPLANT? I’M A MUSLIM! stem cells. But that’s still in the future. In an interview in
2005, leading stem cell researcher Christine Mummery
said that she wouldn’t bet all her money on stem cell
research being successful in this area. “If you need
whole organs, xenotransplantation will be the only
option for a long time to come”, she predicted.
Yang and Sykes (Yang & Sykes, 2007b) say in their
review article that considerable money and effort is
being invested in alternatives to xenotransplantation.
Artificial organs (Textbox 12.5) and mechanical devices
may offer a potential solution for some organ failures,
No government, minister, doctor or patient would but in the near future they don’t have the potential
care to have that on their conscience. On the other to supplant transplantation as a long-term curative
hand, how can you gain more insight into the risks therapy. Likewise organ and tissue regeneration on

Chapter 12: Xenotransplantation 223

 TEXTBOX 12.5. important people of the twentieth century. In 2004
Willem Kolff . 89
Kolff received similar acclaim in the Netherlands:
he finished in 47th place in public elections for De
Willem Johan Kolff (1911-2009) was a Dutch internist Grootste Nederlander (the greatest Dutch man/
who emigrated to the United States. Kolff was best woman), a list of the hundred most important people
known for his invention of the artificial kidney (in 1942 in the country’s history. The American National
in Kampen) and his life’s work on all kinds of other Academy of Engineering calculated in 2003 that
artificial organs, such as a heart-lung machine in 1956 more than 20 million people owed their lives to this
and the artificial heart in 1957. After 1950 he lived and invention of Kolff. Every year hundreds of thousands
worked as a professor in the US. In 1990 the American of people undergo medical treatment that would not
journal “Life” listed him as one of the hundred most have been possible without his work.

the basis of stem cells is very promising, but according organs. This will obviously vary according to the organ
to Yang and Sykes a good many years of research will in question: the heart has a less complex biochemical
be needed before the clinical phase can start. They interaction with the body than the kidneys or liver. But
therefore conclude that xenotransplantation may well this doesn’t detract from the fact that more research is
be the current solution for the lack of donors. definitely needed in the area of xenotransplantation,
however much the opinions of opponents and
This chapter has demonstrated that xenotransplantation proponents differ. Maybe other options, such as organ
is still a tricky business, both technically and ethically. breeding using stem cells, will catch up with the “spare
The technical problems around rejection can probably pig parts” possibility before it comes into practice.
be overcome in the relatively short term, but we This is surely more likely now that we have come to
certainly haven’t read or heard the last word on the the end of the Bush era, when ethical considerations
ethics and the risk of new viruses. Only when the got in the way of federal financing for most research
most serious rejection hurdles and viral risks have involving human embryonic stem cells. With the arrival
been eliminated, can the practice really show whether of President Obama, change is in the air. Than again,
complete pig organs can fulfil their replacement maybe the xenotransplantation cynics will be right after
function as desired, or whether they will simply act as all: “Xenotransplantation is the future and always will
a short-term transition for the transplantation of human be.” Only time will tell if they are right.

89
www.willemkolffstichting.nl/index.php?phm=1

224 Part 3: Health has limits

TEXTBOX 12.6. instance, the specialists thoroughly investigated the
The “perfect match”! possibility of using a family member as a donor. My
brother seemed to be the best “match” for my sister;
As I (JT) mentioned previously, in mid-1997 my sister in fact, the perfect match. Without thinking twice
suffered acute renal failure in both kidneys when she he donated one of his kidneys to my sister. By the
was just 50. This was followed by a long and miserable beginning of 1999 it was a ‘fait accompli’. The transplant
period, in which her close family repeatedly feared for had proceeded successfully and for more than ten years
her life. After more than a year, her condition stabilised now my sister has been able to lead a relatively normal
and she had to undergo dialysis five times a day at life with minimum use of medication; all because of a
equal intervals. Apart from all the other complaints, this perfect match! It all makes you think differently about the
was hardly an appealing state to be in. So the thought alternatives. Not everyone who’s on a donor waiting list
of a transplant wasn’t far from our thoughts. In the first is fortunate enough to have such a brother!

Chapter 12: Xenotransplantation 225

 12.6. SOURCES Prather, R. S. (2007). Targeted genetic modification:
Xenotransplantation and beyond. Cloning and Stem
Aigner, B., Klymiuk, N., & Wolf, E. (2010). Transgenic pigs for Cells, 9(1), 17-20.
xenotransplantation: selection of promoter sequences Rajotte, R. V. (2008). Moving towards clinical application.
for reliable transgene expression. Current Opinion in Xenotransplantation, 15(2), 113-115.
Organ Transplantation, 15(2), 201-206. Schuurman, H. (2008). Regulatory aspects of pig-to-human
Bruzzone, P. (2008). Religious aspects of organ islet transplantation. Xenotransplantation, 15(2), 116-
transplantation. Transplantation Proceedings, 40(4), 120.
1064-1067. Siegert, C., Van Es, L., & Daha, M. (1996). Het
d’Apice, A., & Cowan, P. (2009). Xenotransplantation: The complementsysteem en de klinische gevolgen van
next generation of engineered animals. Transplant stoornissen. Nederlands Tijdschrift voor Geneeskunde,
Immunology, 21, 111-115. 140, 2268-2273.
Deschamps, J. Y., Roux, F. A., Sai, P., & Gouin, E. (2005). Sprangers, B., Waer, M., & Billiau, A. (2008).
History of xenotransplantation. Xenotransplantation, Xenotransplantation: where are we in 2008? Kidney
12(2), 91-109. International, 74(1), 14-21.
George, J. F. (2006). Xenotransplantation: an ethical dilemma. Tallacchini, M. (2008). Defining an appropriate ethical,
Current Opinion in Cardiology, 21(2), 138-141. social and regulatory framework for clinical
Klymiuk, N., Aigner, B., Brem, G., & Wolf, E. (2010). xenotransplantation. Current Opinion in Organ
Genetic modification of pigs as organ donors for Transplantation, 13(2), 159-164.
xenotransplantation. Molecular Reproduction and Tallacchini, M., & Beloucif, S. (2009). Regulatory issues in
Development, 77(3), 209-221. xenotransplantation: recent developments. Current
Louz, D., Bergmans, H., Loos, B., & Hoeben, R. (2008). Opinion in Organ Transplantation, 14(2), 180-185.
Reappraisal of biosafety risks posed by PERVs in Van Zundert, M. (1998). Varkentje vol reserveonderdelen.
xenotransplantation. Reviews in Medical Virology, Chemisch Magazine, pp. 298-299.
18(1), 53-65. Yang, Y. G., & Sykes, M. (2007a). Tolerance in
O’Connell, P. (2008). The rationale and practical issues xenotransplantation. Current Opinion in Organ
for the maintenance of clean herds for clinical islet Transplantation, 12(2), 169-175.
xenotransplantation. Xenotransplantation, 15(2), 91-92. Yang, Y. G., & Sykes, M. (2007b). Xenotransplantation:
Pierson III, R., Dorling, A., Ayares, D., Rees, M., Seebach, current status and a perspective on the future. Nature
J., Fishman, J., et al. (2009). Current status of Reviews Immunology, 7(7), 519-531.
xenotransplantation and prospects for clinical
application. Xenotransplantation, 16(5), 263-280.

226 Part 3: Health has limits

13 THE HUMAN GENOME PROJECT

“The probability of life originating from accident is comparable to the probability of the Unabridged Dictionary
resulting from an explosion in a print shop.”

Albert Einstein90

Charles Robert Darwin, author of On the Origin of Species, was born on 12 February 1809, which was why so much
attention was paid to the creator of the evolution theory in 2009, and why that year was designated the International
Year of Darwin. It was also a good excuse for supporters of Darwin, for creationists (people who believe in the
biblical version of the origin of the world), and other religious believers, to revive the age-old discussion on the
question of whether it is possible as a scientist to believe in God and the Bible, the “book of life”. As the quote above
suggests, even a great scientist like Albert Einstein wasn’t an unconditional proponent of evolution theory, of the
theory that everything just happened by blind chance. The sublime example of ‘the natural order and precision’ is
the DNA, the genetic material in the nucleus of living cells.

LIFE ORIGINATING FROM ACCIDENT IS AS LIKELY AS CREATING A DICTIONARY

FROM AN EXPLOSION IN A PRINT SHOP ...
... RESEARCHERS TAKE PROBABILITY CALCULATIONS EVEN FURTHER!

NATIONAL
LIBRARY

90
answers.yahoo.com/question/index?qid=20071020041348AAmsi18

J. Tramper, Y. Zhu, Modern Biotechnology – Panacea or new Pandora’s box? 227

DOI 10.3920/978-90-8686-725-7_13, © Wageningen Academic Publishers 2011
 13.1. THE HUMAN GENOME 12.2 ‘THE BOOK OF LIFE’

The collection of DNA in a cell nucleus, which is At the very end of the 20th century the world witnessed
contained in 23 pairs of chromosomes in the case of an exciting race between the Human Genome Project
humans, is called the genome, and is regarded by some - an international consortium of scientists from the
as the blueprint of life, the language of God, or in other public sector - and the commercial company Celera
words: “the other book of life”! The study of genomes Genomics headed by Craig Venter. Who would be the
is called genomics. The year 2001 is recognised as the first to decipher the human genome? On 26 June 2000
year in which researchers succeeded in sequencing both parties jointly revealed, with much celebration
all the building blocks in the human genome (Section and in the presence of the former American president
13.3). It consists of two sets of three billion building Bill Clinton and the former British Prime Minister Tony
blocks and there are four different building blocks, as Blair, that they had unravelled the genetic code - five
we saw in the second chapter in Textbox 2.1. Only two years earlier than expected. Prematurely, in a sense,
percent of the genome is used for our 23,000 genes, since neither had done much more than produce a
which contain the code for constructing our proteins, rather rough draft. On 13 February 2001 there was
the building blocks of life. The surprises (see Section yet more ceremony: both draft versions of “the other
13.6) that this unravelling revealed, teach us primarily book of life” were published. Nature, the leading
that the wise old philosopher Socrates was right all British scientific journal, devoted one hundred and
those centuries ago when he said, “The more you fifty pages to it, including annotations. The article had
know, the more you realise you know nothing.” 273 authors and contained the results of the Human
Genome Project, directed at that time by Francis
Collins. The American equivalent Science used
The
up more than one hundred pages to print the data
blueprint delivered by Venter’s company, which began later
of than the public project, but caught up within a year.
life The two most prominent genome hunters, the deeply
religious Christian believer Collins and the aggressive
entrepreneurial scientist Venter, shared the honour.
More than five years later both published a book in
which they examined the human genome from their
own personal perspective.
In The Language of God: a scientist presents evidence

228 Part 3: Health has limits

for belief, Collins (2006) explains why it is that, in his 2008 by Hub Zwart (2008) (Understanding the Human
eyes, science and faith should be able to walk the same Genome Project: a biographical approach). The two
path without conflict. He believes that creationists are questions addressed:
denying truths that science has demonstrated. The 1. What may we learn from these autobiographical
essence of his argument is that the Big Bang and sources about the dynamics of scientific change?
Darwinian evolution theory are sufficient to explain the 2. What is their added value in understanding science
creation of our world, including nature, the human body in general and the Human Genome Project in
and the world of modern molecular biology in all its particular?
complexity, albeit with one exception: our perceptions For non-philosophers the answers are not easy to
of right and wrong. These perceptions can only be understand and to summarise to a length fitting this
explained, according to Collins, by accepting that there chapter and we refer therefore to the original paper.
was a second moment of creation. He believes that What we do understand, though, is the answer of
the language of creation can be read in the genome Francis Collins to the question at a press conference
and in mathematics, two products of two moments of in San Francisco, February 2001, whether the
creation. The book is not an autobiography, but does sequencing warranted the Nobel Prize. He replied that
contain personal anecdotes. it would have to be given to 3492 people to properly
In contrast, Venter’s book, A Life Decoded. My recognise everyone who had significantly contributed
genome: My Life, is a full-length historiography (Venter, to this common effort. Zwart: “Although somewhat
2007). On the one hand he discusses in layman’s rhetorical, no doubt, autobiographical documents
terms what sort of information can be retrieved from reveal that there is a kernel of truth in this reply.”
genomes. On the other, he speaks about the actual In 2004 the Human Genome Project was officially
information that was found in his own genome and the wound down, despite the fact that some parts still
(possible) implications of that for his life. Each book hadn’t been mapped. In subsequent years a lot of
is a justification of the path the authors chose, i.e. the gaps have been filled and work is still being done on
path of a believer (scientist) and that of a (scientific) perfecting it. The American Department of Energy
entrepreneur. Two paths in which the human genome (DOE) maintains the website91 which gives a detailed
did and still does play a central role. history of the project and the milestones, objectives
An in-depth analysis of these two (semi) and results. The question remains, what do we really
autobiographies and of a third one, i.e. The common know and understand about the human genome, and
thread: science, politics, ethics and the human genome what can we do with it, particularly in the area of health
by John Sulston with science writer Georgina Ferry as care.
co-author (Sulston & Ferry, 2002), was published in
91
www.doegenomes.org

Chapter 13: The human genome project 229

 DNA analysis
1. In order to determine the DNA sequence, it is first cut into fragments

2. The fragments are cut so that

they overlap with the cut 3. DNA sequencers determine the
fragments of one chromosome base sequence of the fragments

C G A C C T C T A G G C T T A G T

A T C G G A G C G A T T A A C

A A T C C 4. Overlapping ends
are matched

Old method Commercial method

- Split 1 chromosome - Split several chromosomes
into fragments into fragments
- Split fragments into - Determine DNA = CGTAATC
smaller pieces of fragments
- Determine DNA = CGTAATC - Put fragments back
of fragments together in strands
- Put fragments back together
(search for overlapping pieces)
- Put fragments back together
in strands (search for
overlapping pieces)

Figure 13.1. DNA analysis, adapted from Van der Laan (2000).

230 Part 3: Health has limits

 13.3. HUMAN GENOME SEQUENCING be done much faster and much cheaper, and with that
in mind he set up the company Celera Genomics in
The idea of determining the sequence of the bases the late 1990s. Instead of using enzymes to cleave
(A, C, T, G) in human DNA emerged in the mid- one chromosome into fragments, he used the shotgun
1980s in the US and on 1 October 1990 the Human approach to blast several chromosomes at a time into
Genome Project (HGP) officially started. Work in this fragments of a few thousand base pairs. That can be
area was initially limited to the US, funded mainly done by vibrating the DNA to fragments with ultrasound
by the government and a large national health-care or by forcing it under high pressure through a tiny
organisation, but Europe, in particular the UK, and opening. Using a large number of DNA sequencers
Canada and Japan later added their efforts. The these fragments are analysed, after which superfast
mission of the HGP was:
92
computers fit all the pieces of the puzzle together into
the original DNA chains, gratefully making use of the
“To identify the full set of genetic instructions contained knowledge from the Human Genome Project, which
in our cells and to read the complete text written in the is freely available on the Internet. However, though
language of the hereditary chemical DNA.” much faster, there is also the problem that the margin
of error is much bigger. For this and other reasons,
In order to clarify the sequence of the approximately the two competing factions decided to collaborate at
three billion human base pairs, the DNA was first the beginning of 2000. By the end of June 2000 they
cut into fragments. Two methods were used for this thus jointly presented the first version of the human
purpose (Figure 13.1). The slow, painstaking (‘old’) genome.
method which the researchers of the Human Genome
Project opted for, and a “quick and dirty” approach 13.4. A NEW PARADIGM IN HEALTH CARE
employed later by Craig Venter. In the first method one
chromosome was cut into large fragments, each of Just before the turn of the last century, there was not
which was then individually cut into smaller fragments. only a final sprint to map the human genome, but also
The base sequence in these small fragments was a great many Jules Verne-like predictions as to the
determined automatically using a DNA sequencer. By effect modern biotechnology would have on health
cleaving with specific enzymes that very selectively care in the 21st century. Such futuristic forecasts
break up the DNA chains in specific places, with a bit are not unusual with the advent of a new millennium.
of playing around the original sequence can be found. More than a hundred years earlier, in 1895, Lord
The geneticist Craig Venter believed that it could all Kelvin (an Irish-Scottish physicist, regarded as one

92
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Chapter 13: The human genome project 231

 of the most important physicists of the 19th century) Here are a few examples. He expected there to be a
declared that flying machines were impossible. Eight few successful cases of gene therapy in 2010. As we
years later the Wright brothers proved him wrong! In write, in mid-2010, his prediction has come true to a
the same year, H.G. Wells (1866-1946) wrote ‘The certain extent. In Chapter 11 we concluded that there
Time Machine’, in which the main character travelled had been some success with gene therapy, but that
to the year 802,701 to see a world which boasts the awaited revolution still hadn’t taken place. Collins
ideal preventive health care. Based on their timeline also thought that serious discussions would take
(Figure 13.2), Shamel and Udis-Kessler concluded place on the broad application of diagnostics before
in the December 1999 issue of Genetic Engineering fertilisation, in particular on the consequences thereof.
News, that Wells was probably 800,600 years wrong! This would be for the purposes of identifying genetic
Certain that they could not make a bigger mistake abnormalities that may result in disabled children.
than Wells, they made a hypothetical journey into These predicted serious debates took place in the
the future to see what lies in store for us in 2050. Netherlands in 2008, but at the time the subject was
They spend a day looking through the eyes of their embryo selection. The result was preliminary, very
imaginary ‘heroine’, Karen Rich. Textbox 13.1 is our limited legislation on embryo selection in the event of a
shorter fantasy version of what they ‘see’. genetic predisposition for breast cancer.
As for 2020, Collins predicted that various drugs
NA

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will have been developed on the basis of genetic

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mapped, and that clinical research on extending

1800 1850 1900 1950 1975 2000 2025 2050 2075 2100
life will be taking place. By then the analysis of an
Figure 13.2. Timeline for health care and biotechnology, individual genome (DNA passport) will cost no more
adapted from Shamel & Udis-Kessler (1999) than $1000 per person. This latter forecast now seems
over-cautious, since we are virtually at that juncture
Francis Collins was also seduced into making already. For 2030 he also predicted the availability of
predictions at the turn of the millennium (Potera, computer models of the human cell for research, and
2000). Using the unravelling of the human genome the presence of groups opposing technology in general
as his starting point, he even does it in quite a lot of and modern biotechnology in particular that are avidly
detail for the first four decades of the 21st century. protesting against all these new developments.

232 Part 3: Health has limits

TEXTBOX 13.1. my grandfather finally died of at a very late age.
Diary of Karen Rich; 1 April 2059. Hardly feel the nano-needle go in as the 321 diagnosis
begins.
7-8 a.m.: In next to no time, I find out that there is a microscopic
Switched off the alarm clock and switched on the coffee polyp in my intestines and also what the best prophylactic
machine with my voice, as usual. treatment is.
Looked wistfully at my grandpa’s giant mop of hair in the Decide to begin treatment right away.
last photo I have of him and can just about remember Half an hour later and I’m on my way home after a
the genomics breakthrough that made it possible to cure microinjection from the 321 and with a medical cocktail
baldness. in my pocket to drink before dinner.
Put on my ‘NaturalRed’, no-iron cotton clothes and drink Suddenly realise that 50 years ago there was a good
a cup of breakfast that is scientifically tailored to my chance that this polyp would have gone unnoticed until
morning physiology. it was too late.
Spray on a little perfume, also customised for my 6 p.m. – 11 p.m.:
body - it smells of African orchids, and it’s completely Arrive home and choose one of my ‘personal diet TV
biosynthetic. meals’ – carb-rich bread with a fat-free, protein-rich beef-
8 a.m. – 4 p.m.: flavoured tempeh cutlet, a crunchy, vitamin-E enriched
‘Work’ in Lifeco’s park-style factory churning out biotech- salad, and for dessert a delicious chocolate mousse that
related products made from renewable raw materials. lowers my cholesterol and regulates my insulin levels.
4 p.m. – 6 p.m.: Sit back in my relax-and-massage chair to watch a
Visit my GP for an annual check-up. recent holographic film and enjoy my meal.
Give him my MediChip, which he inserts into the fully 11 p.m.:
automated, diagnostic and therapeutic CustomMed While getting into bed, realise that I can’t imagine how
321. life must have been 50 years ago, in 2009 when people
Don’t think I have any health problems and the report had so many health problems and other worries, like
concurs. the credit crunch and Darwinists and Creationists
But just to be on the safe side, decide to wait a few hammering each other over the head.
minutes to let the 321 perform a full diagnosis on a Am grateful to the scientists who, with a little help from
fragment of tissue sample. modern biotechnology, were able to make the world a
Have a DNA passport that links up to several of the 5000 healthier place.
known genetic abnormalities that increase the risk of a Just before I nod off, I wonder what life will be like in
certain disease; these include intestinal cancer which another fifty years, in 2109.

Chapter 13: The human genome project 233

 Finally, Collins predicted that by 2040 health care 13.5. WILL THE NETHERLANDS CLIMB
will be much more extensive thanks to knowledge ON THE BANDWAGON?
about our genome. Predisposition for diseases will
then be established by looking at individual DNA The decoding of the human genome was, as mentioned
passports and personal preventive health care will above, a thrilling race between the entrepreneur Craig
be available and effective. The testing of neonates Venter on the one hand, and a consortium of mainly
for predisposition to diseases in later life will also be American and British scientists on the other. The rest
possible, although it will not yet be possible to take of Europe, including the Netherlands, stood on the
into account all the environmental factors. In addition, sidelines, but when Clinton and Blair presented the
gene therapy and gene-based medicines will be map of the human genome in a joint show with Collins
available for most disorders and the average lifespan and Venter, the Dutch government suddenly seemed
will be 90. International tension will increase due to wake up. In mid-2000, at the government’s request,
to socio-economic inequality in terms of access to industry and universities wrote the Strategic Action
medical treatments. There will also be debates on the Plan for Genomics. This plan made recommendations
classification of human traits and characteristics. This for investment in DNA research. A year later a follow-up
last reeks of eugenics, the extremely controversial committee delivered concrete proposals and advised
theory that the human race can (or should) be the cabinet to invest € 270 million in genomics over the
improved by selecting individuals with ‘desirable’ next five years, on the basis that the investments should
characteristics (e.g. good health, beauty, intelligence, contribute to the improvement of the quality of life of the
etc) before reproduction. population. Investments should therefore be focused
If we take all the predictions together, the general on the relationship between diet and health, methods
expectation seems to be that there will be a paradigm for improving food safety, mechanisms of infectious
shift in health care in the course of the 21st century. diseases, the occurrence of disorders influenced by
At the moment we go to the doctor when we feel both genetic and environmental factors, and on the
unwell. He makes a diagnosis and prescribes a functioning of ecosystems and sustainability - focusing
treatment. The expectation now is that there will on environmentally friendly and healthy plant and
come a time in the 21st century when not only can animal products. Another interesting recommendation
an individually ‘preventive service book’ be drawn up was to develop objective information so that individual
on the basis of our personal DNA passport for serious members of the public could come to a more balanced
and less serious ‘events’, but we will also be following view. Activities in those five years would prove whether
a personalised ‘preventive diet’. All with a view, of enough had been done to catch up with the top groups
course, to a long and healthy life! in this promising area, where the requisite investment

234 Part 3: Health has limits

seems astronomical, but where the profit can be even about the unexpectedly small number of human genes
greater if the above-mentioned expectations can in compared to the number of human proteins: “So the
any way be realised. And after those five years it would correlation between genes and proteins is really small,
also be demonstrated whether people had a more there’s no debate about that now. We still know too
balanced view on these matters. One thing is sure, little about the expression levels of DNA, RNA and
the Netherlands Genomics Initiative (NGI), which was protein to be able to say anything intelligent about the
set up in 2003, has put the Netherlands back on the correlation between them. There are many preceding
genomics map. This initiative began a second five-year regulatory steps. With every publication I am again
period in 2008 with the help of another € 280 million surprised about how complex the working of genes is.
of government money . Whether this has all helped to
93
And this was not the only surprise that came with the
create a more balanced public opinion is less certain. unravelling of the genome.”
We’ve certainly not been aware of it.
The Dutch contribution to the analysis of the human 13.6. THE SURPRISES OF THE GENOME
genome was therefore fairly minimal in the first draft
versions. The person who was most involved was On the back of the above-mentioned publications in
Gertjan van Ommen, head of the Department of Nature and Science, the Dutch magazine BIOnieuws
Human Genetics at the Leiden University Medical dedicated most of its edition of 3 March 2001 to the
Centre, and founder of the Leiden Genome Technology unravelling of the human genome. The “ten surprises
Center. He was chairman of the Human Genome of the genome” were also included in this edition:
Organisation, or HUGO , not to be confused with
94

the Human Genome Project. HUGO had no direct 1. The genome is like a wasteland. The genome is not
role in the final phase of the Human Genome Project. a nicely arranged row of genes. Extensive “deserts”
Van Ommen: “As far as that was concerned, we at of millions of bases where nothing seems to happen,
HUGO simply stood on the sidelines and applauded.” are alternated by densely populated “urban areas”
HUGO is an ongoing concern with 1,200 members of genes.
around the world and existed before the “sequencing” 2. We have a lot fewer genes than expected, approx.
began. The organisation is primarily concerned with 31,000 (since recalculated at 23,000). Earlier
the dissemination of technologies, patents, ethical estimates started at 100,000 genes or more, but
aspects, name allocation and gene mapping. During those figures were not based on the whole genome.
an interview in 2001, Van Ommen said the following 3. The human genes can make three times more
proteins per gene than the genes of a simple
93
www.genomics.nl organism. Human genes are often built up of little
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Chapter 13: The human genome project 235

 pieces of separate DNA, exons, which can encode 9. There is no scientific basis for racial distinction. At
different proteins in various combinations (Textbox the DNA level all people are 99.9 % alike.
13.3). 10.The genome already offers benefits for health care,
4. The architecture of many human proteins is more for medicine and for the development of medications.
complex than that of a simple organism. Many Genes responsible for human diseases that had
human proteins are multifunctional tools, with not yet been found, are now being discovered via
several active components - the domains. known DNA data.
5. More than 200 human genes are the result of
horizontal transfer of bacteria. These homologues The most important outcome is a paradigm shift in
don’t occur in fruit flies, roundworms or yeast. The our genome knowledge (Textbox 13.2). Knowing,
genes appear to be good at self-preservation or for instance, how so few genes can encode such
even enter into a symbiotic relationship with the a complex organism; after all, humans have only
host. approximately twice as many genes as, for example,
6. Self-replicating base sequences store about a fruit fly, while worm, sand rocket and rat have almost
half of our DNA. This DNA is a fossil archive that the same number. Suddenly the concept of one gene,
enables us to look back almost 800 million years in one protein becomes much too oversimplified, at least
evolution. in the case of humans.
7. There is also ‘junk’ DNA which appears only to
exist for its own sake. For molecular biologists this 13.7. WHERE ARE WE NOW?
is the irritating selfish DNA, such as the segment
christened Alu. This piece of DNA, consisting of The ultimate goal of the Human Genome Project
200 to 300 bases, occurs a million times in the was to construct a map ‘portraying’ all human genes,
human genome. MUTATION SPEED ... i.e. mapping the entire sequence of all base pairs
8. The mutation speed is two (the building blocks) of all the genetic material - the
... ANOTHER THING
times faster in men than MEN ARE BETTER AT! DNA - of humans. A large part of the sequencing
in women. This means was determined in ‘draft’ form for the publication in
not only that the majority 2001. Careful verification was still necessary to rectify
of mutations occur in men, multiple mistakes and it was precisely these mistakes
but that the latter are also that rendered a correct sequencing analysis extremely
unconsciously responsible for complex. Even then there were still large gaps in the
evolutionary changes. sequence that remained undetermined. Attention was
primarily focused on gene-rich pieces of chromosomes,

236 Part 3: Health has limits

TEXTBOX 13.2.
DNA
Paradigm shift: one gene = one protein → one
gene = several proteins.

DNA contains information in the form of genes

which tell a cell which proteins to make (Figure
13.3). Proteins regulate all the processes in our
body. Proteins themselves are made up of smaller
units, called amino acids (20 different kinds), which RNA
are present in the cell. In order to be able to make G A T
A

proteins, first of all a copy (transcription) is made

C T T A G C A T G G A C A T T C

of a small part of the DNA, a gene. This transcript G A A T C G T A C C T G T A A G

G A T

is called messenger-RNA (mRNA) and is in fact an T

T
C T A G

anti-copy, a sort of mirror image of the gene, that is to

C

say mRNA is complementary to the gene. Both ends

of the complementary DNA strings are different. One
Protein
side ends with a phosphate group (see also Figure
1.1 in Textbox 1.1) and is called 5’-terminus. The
other side that ends with the ribose pentagon is the
3’-terminus. Formation of mRNA can only start at the
3’-terminus, so the reading of the gene in the DNA
string takes place in the direction from 3’ to 5’. During
the reading the DNA helix opens and closes as if it
were a closed zipper with two fasteners that move
at a close distance from each other in the same
direction and open and close the zipper respectively.
At the end of 2008 Nature published an article about
a genetic analysis which appeared to show that
not one but several proteins could be made from
nearly all human genes. This occurs via an interim
step, between gene and mRNA (Figure 13.4). First
a temporary mRNA is made, then sections, introns, Figure 13.3. DNA → RNA → Protein.

Chapter 13: The human genome project 237

 A Gene B from that are cut out (spliced). An intron is a piece of
the gene that does not contain code for the pertinent
protein. The code for pieces (domains) of the protein
can be found on the so-called exons. As the figure

Exon1 Exon2 Intron1 Exon3 Intron2 Exon4

shows splicing can be done in various different ways.
(E1) (E2) (I1) (E3) (I2) (E4) The mRNA can also be spliced at a different place
Transcription (alternative polyadenylation). The net result is that
sometimes a fragment of gene is passed over, while
E1 E2 I1 E3 I2 E4 Temporary or pre-mRNA others have two different start or end sites, etc. This
is how two or more active proteins can be created. A
Splicing of I1 and I2 Splicing and exon-skipping
close-up of fragments of DNA around the genes (A

E1 E2 I1 E3 I2 E4 E1 E2 I1 E3 I2 E4
and B) shows other codes which researchers suspect
regulate the amounts of the various proteins. The
proportions can vary dramatically between different
tissues. This orchestration of these processes
E1 E2 E3 E4 E1 E2 E4
ensures that a heart becomes a heart, and a brain
2 variaties of final mRNA
becomes a brain, etc.
Figure 13.4. One gene is translated in several different ways into
messenger-RNA, adapted from Van Santen (2008).

the so-called euchromatic segments, while the gene- Genome Project officially ended in 2004, our genome
poor segments (heterochromatic) have received little had still not been completely mapped. The burning
attention so far. And yet these latter segments also question is whether a final version, in which 99.9% of
contain important structural and regulating elements, the human genome has been determined with extreme
for example the genes that play a key role in the accuracy, will ever come to pass. The relatively difficult
earliest embryogenesis. and therefore expensive mapping of the last segments
The first “draft versions” of our genome therefore still also provides little extra information, while the really
contained a great many gaps and uncertainties. Yet challenging and essential work has really only just
doctors, geneticists and pharmaceutical companies begun, i.e. the search for the significance and effect
were able, for example, to start researching the of all these pieces of DNA. How do you link a piece
genetic causes of some diseases or developing new of DNA to a function in the body, to a disease, to
therapies and medicines for them. When the Human an abnormality? And what happens if, because of a

238 Part 3: Health has limits

mutation, for example, there is an incorrect base pair Other studies from that time also corroborate this.
in the sequence? Only when we have the answers In addition, in May that same year, there was an
to these sorts of questions can we make full use of announcement about the unravelling of the complete
the code of the human genome and can the above- genome of another well-known person, i.e. James
mentioned predictions come true. The explanation and Watson - who together with Francis Crick won the
study of life processes based on the complete set of Nobel Prize for Medicine for discovering the double
an organism’s genetic information is called genomics. DNA helix, which elicited Salvador Dali to say: ‘The
This is, therefore, the area where attention must be announcement of Watson and Crick about DNA … is
focused in the coming decades. Since the Human for me the real proof of the existence of God’ (Stent,
Genome Project ended, great progress has been 1974). By the end of 2007 it had been established that
made in answering some of these questions. Below one individual can easily differ from another individual,
are a few examples in brief. or from something like the above-mentioned average
During the race between the Human Genome Project reference genome, by as many as 15 million of the 3
and Celera Genomics, the researchers from both billion bases, i.e. not 0.1 but 0.5 % genetic variation.
teams combined samples from various individuals, for This discovery of the many major genetic differences
reasons of time and money as well as privacy issues, between individual humans was hailed by Science as
to create a sort of “reference” encoding strand, so that the scientific breakthrough of the year 2007.
they only needed to analyse half, i.e. 3 billion, of the As more genomes are unravelled, the question of
bases. They did so on the assumption that little detail protecting privacy will come further to the fore (Cohen,
would be lost as a result, since the genetic variation 2007). Watson, for example, didn’t want the status of
between different genomes was estimated to be no the key gene for Alzheimer disposition in him to be made
more than 0.1 percent. However, an article published known. Venter, in contrast, laid himself genetically
in 2007 suggested that this was incorrect. In that article bare. In the PLoS Biology article (Gross, 2007) there is
the DNA sequence of both sets of chromosomes of one even a list of more than 20 of his gene variants that are
person was fully described. The genome was that of associated with an increased risk of alcoholism, anti-
none other than the gene hunter Craig Venter himself. social behaviour, tobacco and other addictions, heart
Together with colleagues from the J. Craig Venter disease and Alzheimer’s. Venter is very relaxed about
Institute and three other universities they revealed in making his genome public. He stresses that in the great
the October 2007 issue of PLoS Biology (Gross, 2007) majority of cases, genetic features and diseases are
the sequence of all 46 chromosomes, that is, the set not determined by one single gene. The more people
from his father and his mother. A comparison of these that have their complete genome, characteristics and
two sets reveals that there are many major differences. health status revealed, the easier and more reliably

Chapter 13: The human genome project 239

 TEXTBOX 13.3. characteristics and things like hobbies and favourite
The Personal Genome Project. food and TV programmes. The project wants to
speed up genetic research, in part by giving scientists
In October 2008 Harvard University Medical School access to the information. The volunteers can decide
announced the start of the Personal Genome Project, for themselves whether they want their information to
the final objective of which is to analyse the gene-rich go public. The scientists behind this project hope not
part that codes the proteins, the exome, in thousands only that this will be a quicker way to find correlations
of volunteers, and to put it online . The project began
95
between DNA sequences and specific characteristics,
with 10 volunteers and aims to have 100,000 online but also that it will pave the way for genome privacy
exomes accompanied by photos, medical files, legislation.

scientists can interpret the still very enigmatic human number of known genetic characteristics. In fact, this
genome (Textbox 13.3.). Venter: ‘we have nothing to is comparable to what the fast-growing DTC (direct-
fear, and everything to gain!’ to-consumer) market of genetic testing in America is
offering: for a few hundred dollars you can be screened
“A total gene passport is worth nothing”, this is what for a short or long series of genetic characteristics
Edwin Cuppen contended in October 2008 at his by companies with amusing names like 23andMe96,
inaugural speech as a professor of Molecular Genetics deCODEme97 or the less amusing, but cheap,
in Utrecht, the Netherlands. He had had his own Pathway Genomics98. Cuppen didn’t need his whole
genes mapped for this event, and made them public genome mapped to make his point. What he aimed to
during his oration, with the deliberate aim of provoking do was to prevent the sort of atmosphere surrounding
discussion. By his own admission he had to think twice the introduction of GM crops from building up around
before laying his whole “life” open. Especially since DNA profiling. His proposition is that we should use
when you make your own genome public, you are these new techniques to our advantage. His response
also exposing half the genome of your children. Is that to the question of why we need such a gene passport
really what you want? And what if some unpleasant was: “Doctors and insurers won’t need it for many
facts come to light in the process? Cuppen defended years to come. The new sequencing techniques are
the proposition that, after weighing up all the pros of principal importance in the clinical environment. You
and cons, a gene passport is a good development. don’t want a passport of a patient, but a passport of his
Using apparatus in his own laboratory he had his
own DNA, but not his whole genome, screened for a 96
https://www.23andme.com
97
www.decodeme.com
95
www.personalgenomes.org 98
www.pathway.com

240 Part 3: Health has limits

tumours is very useful. With this you can sometimes African. As the entire cancer genome was screened,
make a better estimate of whether a certain treatment mutations were found in unexpected places. According
will succeed or not. It is more efficient, may prevent a to Professor of Haematology Bob Löwenberg of the
lot of discomfort, and is also cheaper in the end.” We Erasmus Medical Centre in Rotterdam, these would
applaud such debates. They are vital if we are to have have been missed if only the known gene regions had
effective legislation, especially concerning privacy been examined.
and insurability. In 2008 a law was introduced in the If you stretch out all the DNA strands of the 23 pairs of
US prohibiting discrimination on the basis of genetic chromosomes in a human cell and lay them end to end,
factors. they will measure two metres. How the cell manages
There is still no 1000-dollar genome as such, but it is to cram this into its minuscule nucleus is still an almost
on the way, requiring just a few finishing touches to complete mystery. It’s a bit like trying to fill a tennis ball
existing methods. In 2007 it was Venter and Watson with a twenty kilometre long rope. What we do know
who had their entire genome unveiled (at a cost of 1 to is that histones play a role here. Histones are proteins
2 million dollars); in 2008 it was the turn of an unknown that enable the DNA strands to lie close to each other,
Chinese and an African. The analysis of their genomes, without getting in a tangle and becoming completely
or at least 99% of the sequence and all the interesting inaccessible (Textbox 13.4). In 2008 another piece of
bits of it, took two months and cost 250,000 dollars, this mystery was solved by researchers at the Dutch
but by the end of 2008 there was one company which Cancer Institute and the Erasmus Medical Centre. Bas
claimed to be able to sequence a human genome for van Steensel and his colleagues wrote in Nature of 7
5,000 dollars. The 1000-dollar genome will offer many May 2008 that long pieces of human DNA strands are
possibilities for cancer treatment. Cancer is a real DNA stuck to the inside of the cell nucleus wall (Figure 13.5).
disease. It is the changes in the DNA of body cells that
Stop code
convert a cell into a cancer cell, and it is these DNA Cell nucleus wall

changes that determine the behaviour of the tumour -

how fast it grows, or spreads and what chemotherapy
it is sensitive to. All this information is stored in the Active DNA Inactive DNA
base sequence of the tumour cell DNA. Affordable
and reliable access to this knowledge may therefore Figure 13.5. Cell nucleus: inactive pieces of DNA glued to the
revolutionise the treatment of cancer. The 6 November nucleus wall, adapted from Rouwé (2008).
2008 issue of Nature contained the first ever description
of the whole genome of a human cancer cell, as well The researchers found 1,300 marked pieces of DNA
as the above-mentioned genomes of the Chinese and that stuck to the nucleus wall. These pieces were

Chapter 13: The human genome project 241

 TEXTBOX 13.4. RNA polymerase can read hundreds of base pairs per
‘Reading genes’. minute, but if the DNA is still wound up, it takes a lot
longer. It may then take a few minutes before there
The protein spools around which the DNA helix are random movements of the molecules in question,
is tightly wound are the histones, which in turn are to give each other space again. Researchers from
very tightly folded up like beads on a string in the the University of California in Berkeley were the first
cell nucleus. In order to be able to function, the cell to record this acceleration and deceleration process
must constantly read information from the DNA. around twisted DNA. They believe that the slowing
Figure 13.5 gives an idea of how a cell does this. down by the histones is necessary for all kinds of
The gene-reading protein RNA-polymerase begins processes that genes and RNA undergo before
by enveloping an unravelled fragment of DNA. It protein synthesis. They published these views in
briefly separates the two strands from each other the 31 July 2009 issue of Science (Hodges, Bintu,
and copies the base sequence to an RNA molecule. Lubkowska, Kashlev, & Bustamante, 2009).

relatively big and strictly limited by stop codes: stick two metres of DNA strands in the cell nucleus (and that
to the wall up to here, and let the rest hang loose. they can still have accessible, and active parts) has at
Remarkably enough, the DNA attached to the nucleus least gone a little further towards being solved. Textbox
wall only contained inactive genes. Active genes 13.4 describes an even later development in this area.
are present in the loose loops and can be read. The
researchers suspect that the cell deliberately makes 13.8. IN CONCLUSION
- and keeps - certain genes inactive by sticking them
to the nucleus wall. They also believe that the cell can “It was premature to describe the human DNA code
move pieces of DNA from the wall to the loose loops as the ‘book of life’”, said Maarten van Lohuizen of the
in the middle of the nucleus, and vice versa, probably Dutch Cancer Institute during his inaugural speech
to activate or inactivate, respectively, new genes under as part-time professor at the University of Utrecht in
new circumstances. According to Steensel, which 2004. “We still have to learn how to decipher another
pieces of DNA are attached to the nucleus wall, differs important foreign language: the histone code.” The
not only from one time to another, but also from one histone code delivers no template for producing
cell to another. For example, it will be different in a skin proteins like DNA. The code simply determines to a
cell than in a liver cell. It is for that reason that they great extent which gene or which set of genes in a cell
are investigating different types of human cells. The is active or not, or which is in a sort of standby position.
puzzle of how one minuscule human cell can contain The picture of DNA as a blueprint of life is therefore far

242 Part 3: Health has limits

too simple. In addition, according to the upcoming field CD: a gigantic collection of data that only has meaning
of epigenetics, the DNA sequence is not the only thing when it is “unlocked”. It is a carrier of information, but
that determines someone’s genetic characteristics. not the essence of it. In his book The Music of Life,
What the mother eats during the pregnancy can also Noble compares the organism and its behaviour to
have an influence. Epigenetics is the study of reversible a musical performance. He interprets this metaphor
genetic changes in gene functions that occur without almost literally, and shows how brilliantly it fits. “Is
changes to the DNA sequence. It is also the study of there a score? No, only a series of molecules. Is there
the processes that affect the unfolding development of a composer? Yes, evolution. And a conductor? Yes,
an organism. In both cases a study is made of how natural law and the organism. What is the orchestra?
gene-regulating information, not expressed in DNA The organs and biochemical processes in the body.
sequences, is transferred from one generation (cells or And everything responds to everything else.”
organisms) to the next. In other words, “in addition to” There’s no doubt that the unravelling of our genome has
the genetic information that is encoded within the DNA. revealed many things and induced many developments
The enormous variety of body cells that we possess, - not only in scientific labs. At the very least it has
e.g. brain cells, heart cells, liver cells, skin cells, etc., taught us that things are a lot more complicated than
cannot be explained by DNA alone. Scientists are we thought, and that we are still at the beginning of
therefore now looking for a sort of instruction booklet, the learning process in many areas. As for answers
that tells each cell type how to read the human genome to questions like “where does chemistry become life,
for itself. where does life become consciousness, and where
In short, anyone who thought that we had reached the does consciousness become individual personality?”,
end of the road when the entire genome was unravelled, they will be a long time coming.
was completely mistaken. Other “languages”, such as June 26, 2010 marks the tenth anniversary of the
histone codes, first need to be deciphered. And even initial decoding of the human genome’s 3 billion base
when we know those, we are probably still only at the pairs, this is how the editorial in The Lancet starts in
beginning of the journey. The interaction between the the issue of that date. The progress made since then
various components of an organism and the dynamics is summarised on half a page and corresponds rather
thereof also need to be learned - a new domain of well with the topics in this chapter. The editorial ends
knowledge known as systems biology. One proponent with the hope of The Lancet that the second post-
of systems biology is the British physiologist Denis genomic decade will progress towards personalised
Noble. He is trying to fathom the interplay between medical care by the application of targeted therapy for
genome, organism and environmental factors at all individual patients, a hope we can fully support.
levels (Noble, 2008). Noble likens the genome to a An article with a somewhat similar title, The View a

Chapter 13: The human genome project 243

 Decade On, as the headline of the editorial in The disease, seems less like a paradigm shift than a new
Lancet, reviews the book Drawing the Map of Life frontier, once again driven by new technologies. The
– Inside the Human Genome Project by Victor K. future trajectory, McElheny suggests, is promising
McElheny . It is written by Angela N.H. Creager in
99
though unpredictable. Drawing the Map of Life
Science of 9 July 2010 and she concludes with: “The sketches out a more complete history of genomics
book’s depiction of current trends in biomedicine, with than previously available, but clearly the story is not
the decline of ‘gene-centered’ accounts of traits and yet finished.” This chapter is!

99
shass.mit.edu/news/news-2010-mcelheny-drawing-map-life

244 Part 3: Health has limits

 13.9. SOURCES Rouwé, B. (2008, 10 May). DNA hangt met zijn inactieve
delen aan de kernwand. NRC.
Cohen, J. (2007). Genomics: Venter’s genome sheds new Shamel, R. E., & Udis-Kessler, A. (1999, December).
light on human variation. Science, 317(5843), 1311- Biotechnology in the 21st century. Genetic Engineering
1311. News.
Collins, F. S. (2006). The language of God: a scientist Stent, G. (1974). Molecular biology and metaphysics. Nature,
presents evidence for belief. New York, Free Press. 248(5451), 779-781.
Gross, L. (2007). A New Human Genome Sequence Paves Sulston, J., & Ferry, G. (2002). The common thread: A story
the Way for Individualized Genomics. Plos Biology, of science, politics, ethics, and the human genome.
5(10), e266. London, Joseph Henry Pr.
Hodges, C., Bintu, L., Lubkowska, L., Kashlev, M., & Van der Laan, S. (2000, 12 August). De geheimen van het
Bustamante, C. (2009). Nucleosomal Fluctuations genoom. Chemisch2Weekblad, p. 31.
Govern the Transcription Dynamics of RNA Polymerase Van Santen, H. (2008, 8 November). Veelzijdige
II. Science, 325(5940), 626-628. boodschapper. NRC.
Noble, D. (2008). The music of life: biology beyond genes. Venter, J. C. (2007). A life decoded: my genome, my life.
Oxford University Press, USA. Viking Press.
Potera, C. (2000, August). Life after human genome map. Zwart, H. (2008). Understanding the Human Genome Project:
Genetic Engineering News. a biographical approach. New Genetics and Society,
27(4), 353-376.

Chapter 13: The human genome project 245

14 STEM CELL THERAPY:
PROMISING AND CONTROVERSIAL!

“Mankind has been forever in search of eternal youth. Where magicians and alchemists failed in their efforts, the
biomedical scientist seems to offer the promise of eternal life with the discovery of the stem cell.”

Hans Clevers and Ronald Plasterk

This is a quote from the foreword of a book called Stamcellen (Stem Cells), written by one of the world’s leading
stem cell researchers, Christine Mummery, and two of her colleagues (Mummery, Van de Stolpe, & Roelen, 2007).
Stem cells are cells that, depending on the conditions, form specific cell types, tissues and organs. They don’t yet
have any specific or specialist function like normal (somatic) body cells, for example blood cells, skin cells and liver
cells.

THE SEARCH FOR ETERNAL YOUTH ...

... MAGICIANS FAILED, SCIENTIST WON!
LOSER!

SHUT UP!

J. Tramper, Y. Zhu, Modern Biotechnology – Panacea or new Pandora’s box? 247

DOI 10.3920/978-90-8686-725-7_14, © Wageningen Academic Publishers 2011
 14.1. HUMAN EMBRYONIC STEM CELLS we subscribe, clearly represents the status of stem
ARE ‘HOT’ cell therapy in the first decade of the 21st century:
promising yet controversial! Afraid of controversy, the
In the earliest stage of a human embryo, immediately Bush government largely restricted activity in this area,
after the first cleavages of the zygote (fertilised egg), but fortunately, with the entry of Obama into the White
all cells are still identical, still undifferentiated and can House, clinical stem cell treatments look within reach
in principle still multiply endlessly and develop under again (see next section).
the right conditions into any of the two hundred and When the first edition of the book “Stamcellen” was
ten differentiated, adult cell types that go to make up published in 2006, Clevers and Plasterk were both
our body. This great capacity of the early embryonic directors of the Hubrecht Laboratory in Utrecht, the
stem cells to differentiate is called pluripotency. centre par excellence for research on developmental
Pluripotency and the endless growth capacity are the biology and stem cell therapy. It is an institute of the
characteristics that are useful in stem cell therapy: KNAW (Royal Netherlands Academy of Science),
the targeted cultivation of a specific sort of tissue cell where Mummery worked for years. Since 1 April
from embryonic stem cells in a laboratory, and the 2008, she has been a professor at LUMC (the Leiden
transplantation of these to a patient, for example heart University Medical Centre). Plasterk left science for
cells to someone who has lost heart muscle tissue politics in 2007, to become Minister of Education.
following a heart attack. Stem cell research obviously is a dynamic field, not
“Embryonic stem cells have achieved prominence in only in the lab. Since the appearance of the stem
part because of the still unsubstantiated hopes that cell book another spectacular and paradigm-shifting
therapies that use them can ameliorate a variety of development has taken place, namely the manufacture
human ailments. They have attracted controversy of human pluripotent stem cells by the induced de-
mainly because the cells are obtained from human differentiation of specialised adult cells, or in other
embryos, linking stem cell research to historical battles words the “reprogramming of differentiated normal
over abortion and over the legal and moral status of cells back into pluripotent undifferentiated stem cells”
the human embryo and fetus.” This abstract is from (Section 14.7). There is a good chance that in the long
the chapter on stem cells in the book “The Art and run this will remove the need to make pluripotent stem
Politics of Science” by Harold Varmus (2009). Varmus cells from embryos, and thus avoid the accompanying
won the Nobel Prize for Medicine in 1989 and was controversy. Despite these rapid new developments,
director of the American National Institute of Health the book by Mummery et al. is still well worth reading,
(NIH), which allocates billions of dollars every year to because it discusses in a clear way the underlying
medical research in the US. His point of view, to which ideas and principles of stem cell therapy. The

248 Part 3: Health has limits

interested Dutch reader can use this as a basis for would, according to him, form the framework within
understanding and interpreting the new developments, which research on embryonic stem cells could evolve in
discussions and publications on this subject in journals a scientifically valuable and responsible way. However,
or newspapers, and thereby establish an informed Tom Okarma, head of Geron, said that the draft
opinion on this hot topic. An updated English version guidelines showed that government officials have still
could in our opinion form a very useful contribution to not fully understood the potential of human embryonic
educate the broader public globally. Anyway, we were stem cells. Given the potential of stem cell therapy, a
very grateful for the use of the book Stamcellen to write much broader framework and much stronger incentives
this chapter. are required. In his view, stem cell treatments will be
able to cure previously untreatable diseases, thereby
14.2. FROM BUSH TO OBAMA saving lives as well as money that is currently spent
on ineffective medicines. Treatments with embryonic
Shortly after President Obama was inaugurated in stem cells are not 100,000 dollar therapies that extend
January 2009, the Californian company Geron was lives by three weeks, says Okarma. With a simple
given the go-ahead by the FDA to start the first clinical intervention you can permanently repair a defective
trials with human embryonic stem cells. Although
the FDA vigorously denied that this had anything
FROM BUSH TO OBAMA ...
to do with the arrival of Obama in the White House,
nonetheless it did mark the beginning of a new era.
Obama’s predecessor, President George Bush jr, had
in 2001 restricted all federal support for research with
1 - 0 FOR ME,
human embryonic stem cells to 15 then-existing, NIH-
MISTER BUSH!
registered, stem-cell lines, the “Presidential list”. More
indications of a new stem cell era quickly followed.
In March 2009 Obama declared that he wanted to
lift the restrictions that Bush had imposed. A month
later, the Obama government laid down guidelines
to regulate this research. In mid-2009 a White House
spokesman announced that the administration was
busy processing approximately 50,000 reactions from
the public on these draft guidelines and that the final
guidelines would be ready by early July 2009. These

Chapter 14: Stem cell therapy: promising and controversial! 249

 function of an organ or tissue that has been damaged Researchers will monitor the patients for over a year
by an injury or disease. According to Okarma what is to find out whether the treatment is safe and whether
needed is a presidential committee of experts to advise defective functions and movement possibilities have
the administration on government policy, allocate been repaired.
research grants, and promote collaboration between If this clinical trial is successful, much of the resistance
researchers in industry and academia. to applications with embryonic stem cells will probably
fall by the wayside. In any case, the new era has
The final guidelines for allocating federal money to started with an explosion of spectacular novelties.
stimulate stem cell research, more than ten billion There isn’t a day goes by without one or two appearing
dollars, were made public on 6 July 2009. Only on the Internet. On 27 July 2009, for example, there
research on stem cells from embryos left over after was a report on the identification of the most suitable
in-vitro fertilisation (IVF) is eligible for federal funding, stem cells for cultivating bone implants and a report
provided there is written consent from the donors. How on surgical thread with embedded own stem cells to
all this will evolve is not yet clear, but Geron began stimulate the healing process of sutured wounds. A
the first clinical trials in early 2009. The trials involve year later, as another example, in the July-August 2010
experimental treatment with embryonic stem cells for issue of Euro/Biotech/News it is reported that Italian
patients paralysed by transverse myelitis (spiral-cord researchers have restored sight to blind patients using
injuries). The study was however halted after seven stem cells from the patients’ own bodies; it concerned
months because safety concerns surfaced in an 106 patients whose eyes had been severely damaged
animal study, which showed an increased frequency by chemical burns. The preceding May-June issue
of small cysts within the injury site in the spinal cord. of this journal brought somewhat unexpected, but
In response, Geron developed new testing methods pleasantly surprising news: “The Vatican has taken
that essentially ensure the purity of the drug, which a bold step into unchartered territory with its decision
is actually a mix of different cells. On 31 July 2010 to finance new research into the potential use of adult
Thomas Gryta writes in the online Wall Street Journal stem cells for the treatment of intestinal disease and
that the FDA has cleared Geron to resume their stem possibly other conditions.” Although the Vatican has a
cell study. The initial testing in humans will focus on positive stance with respect to transgenic crops (see
the safety of the drug, and its effectiveness must still Textbox 3.1 in Chapter 3) and to biotechnology in
be proved. The study will evaluate safety in eight to general, it is on the understanding that there should
ten patients with recent severe spinal-cord injuries. continue to be a ban on cloning humans and tinkering
The company agreed with the FDA to leave 30-day with human DNA (Chapter 1). Nevertheless this news
intervals between the first patients, for safety reasons. seems to be a first step in a slight loosening of the ban.

250 Part 3: Health has limits

 14.3. THE CONTROVERSIES It wasn’t until 1994 that interest in human stem cells
was renewed. In that year an NIH (American National
The current debates on stem cells and the political Institutes of Health) panel issued a report on possible
policy that regulates their use, have been influenced prospects for research into early embryogenesis. It
by several crucial events, whereby the position of was written with the prospect of there being a new,
successive American governments has had a major and in all expectation, permissive political policy. The
impact on what happened in the rest of the world. recommendations for research on human embryos in
Each of these events was of critical significance for this 1994 report were almost identical to recent and
the course of stem cell research. It began in 1978 promising work at that time on mouse embryos and
with the birth of Louise Brown, the first IVF baby. In- stem cells. The report also expressed the expectation
vitro fertilisation is the fertilisation of human eggs in that major progress in mammal biology would
the laboratory and, after a few cell divisions, the result, which would greatly facilitate the successful
implantation of the formed embryo into the uterus, application of human embryonic stem cells in clinical
giving childless couples a greater chance to have a research. Changes in the political climate, however,
child. IVF took off after the birth of Louise Brown. Since led subsequently and all too quickly to a ban which
more eggs were fertilised than embryos implanted, and suspended much of the research recommended in the
there was no possibility of preserving the excess IVF report. In 1996 US congress banned the use of federal
embryos for later IVF use, there was little discussion funds to create or destroy human embryos solely for
about their alternative use. As a consequence and research purposes.
with the minimum of discussion, in Oxford (England) It was Ariff Bongso, a pioneer in the field of stem cell
amongst other places, the remaining embryos were research, who was the first to recommence work on
used in the first, albeit failed, attempts to obtain an human stem cells in Singapore. In 1994 he described
embryonic stem cell line. A cell line consists of the a procedure for removing cells from a blastocyst
same type of cells which can be further cultivated and cultivating them in a petri dish in the laboratory
under suitable conditions in the laboratory. Only when, (Section 14.4). A blastocyst (Figure 14.1) is a small
a short while later, a freezing procedure for embryos round structure filled with fluid and cells, which is
was developed, did the ethical discussions on the use formed after several divisions of the fertilised egg.
of remaining IVF embryos begin, particularly because In mammals, where implantation of the blastocyst
the stock of frozen IVF embryos grew exponentially. takes place in the uterus, the cells which form the
The often heated discussions resulted in the blocking actual embryo are located as a cluster of cells, the
of research on human embryonic stem cells, and inner cell mass (or embryoblast), eccentrically in the
research shifted again to the stem cells of mice. blastocyst.

Chapter 14: Stem cell therapy: promising and controversial! 251

 were all difficult to cultivate and therefore not really
suitable for research or use in applications.

Endometrium
For a long time the use of human embryos for the
creation of stem cell lines was only possible in a
very limited sense in many European countries too.
Inner cell mass Yet, a number of research groups inside and outside
the United States, and especially in Asia, stubbornly
persisted with the development of new human
embryonic stem cell lines and made significant
Trophoblast
advances. The International Stem Cell Initiative
was set up in January 2003, and decided in 2005 to
Blastocyst cavity compare all registered human embryonic stem cell
lines with each other to establish similarities and
differences and to stimulate further research; 75
Figure 14.1. A blastocyst at the endometrium, the mucosa cell lines from 14 countries around the world were
that lines the inner wall of the uterus in which the blastocyst involved in this research. Since 2002 the use of
becomes implanted. human embryos for making stem cell lines has been
permitted in the Netherlands under certain conditions
The American James Thomson used this method in (with the consent of the CCMO (Central Committee on
1998 to make the first human embryonic stem cell line. Research Involving Human Subjects)). A declaration
This scientific breakthrough breathed new life into of consent from the donors of the embryos (both the
stem cell research and in no time various laboratories man and the woman) is a prerequisite. Since July
were isolating new human stem cell lines. This 2009 that has also become a crucial precondition in
research has been politically and ethically charged the US. However, because of the restrictions many
from the beginning, and boycotted to a large extent research groups have shifted their attention to less
by the Bush government. As previously mentioned, emotionally charged research on adult stem cells.
in 2001 President Bush banned government funded Stem cells occur not only in embryonic tissue, but in
research with human embryonic stem cell lines which virtually all our tissues. Admittedly, these adult stem
were made after 9 August 2001. This effectively cells have more limited differentiation possibilities,
restricted stem cell research in the US thereafter to but this field has nevertheless advanced in leaps
the ‘presidential list’, the fifteen stem cell lines which and bounds. After years of fundamental research,
were officially registered at the NIH; these cell lines these adult stem cells are now beginning to bear

252 Part 3: Health has limits

fruit in medical applications100. In December 2008 the specialised human cells back to the stem cell stage.
Translational Adult Stem Cell research programme By the end of 2007, it had happened: the era of the
began in the Netherlands. It had a budget of more “formation of induced human embryonic pluripotent
than 22 million euros for research into making stem stem cells by dedifferentiation of specialised cells”
cell therapy a reality for patient care. had arrived, and led to feverish new developments
The final crucial event was the birth of Dolly the Sheep without the ethically-charged label carried by stem cell
in 1997 - a remarkable scientific achievement. The lines isolated from human embryos (Section 14.6). It
way in which biologists looked at the arrangement went hand in hand, in the US too, with a relaxing of
of genetic information changed fundamentally as a the restrictions under which human embryonic stem
result. Up till then it was thought that the transition cell research could be carried out. For example, at
from undifferentiated stem cells to fully differentiated, the end of 2004 a referendum in California resulted
specialised tissue cells was effectively irreversible. in the release of three billion dollars for embryonic
All body cells do have precisely the same genome, stem cell research. The Californian Institute of
but in stem cells there is a completely different Regenerative Medicine was set up, but the money
package of active genes from those in specialised only became available in mid-2009 after delays
cells. After many futile attempts, the introduction caused by lengthy legal procedures. Within four
of the genetic material from an udder cell from the years, from the beginning of December 2009, the
“mother sheep” into an oocyte of another ewe and institute hopes to have ten to twelve new stem cell
the implantation of the formed “embryo” in the uterus therapies in the clinical trial phase with humans.
of a third, resulted in the clone Dolly. It showed that, The aim of the institute is to promote the transition
in contrast to expectations, genetic reprogramming from tests performed on animals in the laboratory
of adult specialised cells to much earlier stages in to tests in humans in the clinic, very like the Dutch
their development is a very real possibility. It also Translational Adult Stem Cell Research programme.
suddenly opened the way to ‘patient-specific’ stem It looks therefore as if, in the early days of the Obama
cells for personal therapy. But Dolly’s birth also raised era, the lines have been redrawn. Time will provide
fears of human reproductive cloning, which seriously the answers to the questions whether stem cells are
limited any desire to design a promising method for ultimately suitable for therapeutic applications and for
reprogramming cells for therapeutic purposes. So which disorders, but primarily with which stem cells:
it was more than ten years before the publication embryonic, adult, or induced pluripotent stem cells, or
of the first examples of genetic reprogramming of perhaps all three?

100
www.xcell-center.com

Chapter 14: Stem cell therapy: promising and controversial! 253

 14.4. WHAT IS A STEM CELL (THERAPY)? As we have seen, stem cells in a much earlier stage
of human and animal development, i.e. in the early
The term “stem cells” has a well-defined meaning for embryo, have much greater potential. These early
biologists, and implies more than just the controversial, embryonic stem cells are the progenitors of all 210
politically-charged types originating from human (differentiated) cell types from which tissues and
embryos. All specialised cells such as skin cells, organs of the adult human are made. It is because of
liver cells and brain cells are formed in humans and this “plural” potential that they are called pluripotent.
animals by means of an orderly process, in which In principle, pluripotent stem cells offer the most
undifferentiated cells divide and differentiate into possibilities for stem cell therapy.
these specialised cells. Under certain conditions, the Stem cell research is to the first decade of the 21st
undifferentiated cells can form two types of daughter century what recombinant DNA research was to the
cells when they divide: a daughter cell that cannot be 1970s and 1980s, and the Human Genome Project
differentiated from the mother cell and does the same was to the 1990s - the most visible and most striking
thing, and another daughter cell that moves towards manifestation of the promising and spectacular
becoming the specialised cell. These undifferentiated developments in the biological sciences. Most stem
cells, which not only endlessly replicate but can also cell biologists agree that human embryonic stem cells
produce differentiated offspring, are called stem cells. have the greatest potential in principle to treat human
Many stem cells also occur in adult tissue and have a diseases and wounds (Gruen & Grabel, 2006). This
limited ability to differentiate, for example, blood stem expectation is based on the observation that these
cells differentiating into all sorts of blood cells such as stem cells can differentiate themselves into most, but
red and white blood cells, but not into insulin-producing not all, cell types from which the adult human body is
cells. Such “multipotent” adult stem cells can divide in composed, not only in the body (in vivo), but also under
two ways (Figure 14.2). suitable conditions in vitro (test-tube, Petri dish, etc.).
The road to successful stem cell therapies therefore
seems to be a straightforward one: make the desired
specialised cell type from human embryonic stem
cells, for example, pancreas β-cells for the treatment of
diabetes type 1 (an autoimmune disease whereby the
patient cannot make insulin in the body), and transplant
these cells to the desired location in the patient. This is
Figure 14.2. The two ways in which adult stem cells can easier said than done, though, because the knowledge
divide, adapted from Mummery et al.(2007). and technology required for such a process has not yet

254 Part 3: Health has limits

been sufficiently developed. There is an abundance of form of serum, and/or additives and growth factors.
scientific literature on differentiation and transplantation Cells to be transplanted must be guaranteed to be
of mouse stem cells, but the literature on differentiation free of animal substances which contain pathogens or
and transplantation of human embryonic stem cells is cause immune reactions. For that reason, researchers
lagging far behind. However, the distance between the are eagerly looking for effective purification methods
two is diminishing all the time. Nonetheless, a great for the existing cell lines and investigating isolation
many hurdles, both ethical and scientific, still need to techniques and cultivating methods that don’t use
be overcome before we see the routine application of animal components. The risk of transplanted cells
stem cell therapy in the clinic. themselves being rejected is also very real. Since cells
One hurdle that is not often explicitly mentioned is the that are genetically identical to those of the patient are
resistance, not to say aversion, which a great many the most promising approach to this problem, intensive
researchers have to investing time in acquiring a research is under way to find methods for making these
better understanding of the nature of possible ethical “patient-specific” cells. Finally, there is an obstacle that
resistance to their work. It is usually these researchers has become much greater in recent times, namely
that are best suited to feeding the ethical and political the funding of stem cell research. Barack Obama has
debates with relevant and objective information, opened the doors for this research again and hopefully
providing that they have the right skills to (dare to) where he leads others will follow. It will certainly help
discuss and defend their work on ethical grounds. In scientists to overcome the other obstacles.
our view, universities could play an important role here
by giving the subject more attention and form. 14.5. TYPES OF STEM CELLS
An important scientific obstacle yet to be tackled is
the risk of tumour formation, which accompanies An investigation of the development of a fertilised egg
every transplantation with undifferentiated cells. Many into an adult human presents a good picture of the
methods are being researched to eliminate the tumour- various different stem cells that exist (Figure 14.3).
forming cells, as far as possible, before and after The merging of a sperm and oocyte creates a zygote,
transplantation. As with any experimental therapy, an whereby the sperm cell is “swallowed up” by the much
acceptable level of risk must be carefully defined. bigger ovum. A male and female set of chromosomes
What is essential here is that the patients eligible for come together and fuse into one cell nucleus which
treatment are given as much information as possible contains the whole genome, i.e. in the case of a human
on all aspects of their treatment. Until recently, nearly cell nucleus 23 pairs of chromosomes. The cytoplasm
all human stem cell lines were isolated and cultivated contains the necessary components for this fusion, but
in the presence of animal components, often in the also for the first cell divisions. Approximately one day

Chapter 14: Stem cell therapy: promising and controversial! 255

 after fertilisation the human zygote cleaves itself into a After about three days and three cell divisions a
two-cell embryo, then into a four- and eight-cell embryo solid cluster of eight blastomeres is formed, called
(cleaving divisions), whereby the original cytoplasm is morula (mulberry). From this stage onwards, the first
distributed over all the cells, so that there is no change morphological (shape and composition) differences
in the total volume. The cells of these early embryos between the cells can be observed and the totipotency
are called blastomeres and in theory each is able, rapidly decreases. After four days the blastocyst stage
individually, to form a complete embryo that can nestle is reached, and more than a day later the blastocyst
into the lining of the uterus and develop into a complete (Figure 14.1) implants itself into the uterus. There are
individual. This property is called totipotency. only two cell types in the early blastocyst, namely the

Blastocyst

Zygote Morula

Human fetus

Totipotent

Pluripotent

Ectoderm Mesoderm Entoderm Reproduction cells

Nerve cell Skin cell Bone cell Blood cell Pancreas cell Loung cell Sperm cell Ovum

Figure 14.3. From zygote to fully specialised cell.

256 Part 3: Health has limits

trophoblasts and those in the eccentric cluster of cells, have all the properties of the final fully matured cell.
the inner cell mass. The latter is the actual embryonic These progenitor cells are called unipotent, because
part which will later form the fetus and the new they can only differentiate into one cell type. Yet
individual. The so-called extra-embryonic structures, they might still be interesting for cell transplantation,
such as the placenta and the umbilical cord, are not because in principle they can still multiply outside the
formed from the cells of the inner cell mass, but come body, albeit to a limited extent. The final completely
partly from the trophoblast cells which surround the differentiated cell has to exercise its role within the
fluid-filled cavity and partly from the eccentric cluster of organ and can generally not divide or only to a limited
cells of the blastocyst. Since all cells of an adult human, extent. However perfectly it functions, such a cell is no
except those of the extra-embryonic structures, can longer suitable for cell transplantation purposes.
be created from the inner cell mass, they are called
pluripotent. 14.6. THE MAKING OF HUMAN (EMBRYONIC)
The pluripotent stage doesn’t last long, because the STEM CELL LINES
cells rapidly differentiate during the normal embryonic
development into more specialised cells. In addition The making of a cell line involves the isolation of
to the progenitors of the reproductive cells, three new a certain type of cell from a tissue or organ and the
cell types emerge from the inner cell mass. These cultivation thereof, so that only this type of cell appears
are called cotyledons: the outer layer or ectoderm, in the culture. A considerable number of cells can be
the middle layer or mesoderm and the inner layer or cultivated from this cell line and frozen in small portions
endoderm. All human organs and tissues stem from at very low temperatures. These frozen cells constitute
these. A few examples of specialised cells that are the cell bank with which further work can be done
formed from the various cotyledons are given in Figure over a long period of time. The standard procedure
14.3. Stem cells are still present after the embryonic for isolating stem cells uses embryos in the blastocyst
stage, but they have a more limited potential. In most, stage. The cells of the inner cell mass are isolated from
and perhaps even in all, organs and tissues of an adult the blastocyst and spread out on a special medium in
individual there is a small stock of adult stem cells. Petri dishes. The development of a suitable medium on
Such organ or tissue stem cells can still divide and which the stem cells can multiply without differentiating,
generally develop into a limited number of cell types requires specialist knowledge, expertise and research.
of which the tissue or organ in question consists: they It is this approach (Figure 14.4A) that raises the ethical
are multipotent. objection, i.e. that embryos are lost in the process.
Differentiating multipotent stem cells give rise to A second method (Figure 14.4B), to which there is no
progenitor cells which can still divide, but don’t yet objection in principle, involves isolating a single cell, a

Chapter 14: Stem cell therapy: promising and controversial! 257

 A: Standard procedure

Cell line
8-cells phase

Blastocyst

B: Cell line from single blastomere

Implantation in uterus

C: SCNT (= somatic cell nuclear transfer)

D: Alternative SCNT

Somatic cell Implantation in uterus

cdx2-/-

cdx2-/-
E: Cell fusion

x =
Somatic cell Embryonic stem cell Hybrid cell

Figure 14.4. Five ways to make a human embryonic stem-cell line, reproduced with permission (Gruen & Grabel, 2006).

258 Part 3: Health has limits

blastomere, from an even earlier stage of the embryo. is possible to start from one cell. It is a well-known fact
The cell in question is one of the eight fourth-generation in animal cell culture that a minimum number of cells
cells (1 → 2 → 4 → 8) which come into being as a are necessary for growth. This cell layer, the so-called
result of 3 cleaving divisions. The isolated blastomere feeder, gives off a factor which prevents the cells from
is spread (plated out) on a medium which already differentiating and losing their pluripotency; in 1988
contains a cell layer. Normally speaking many cells are this factor was identified as leukaemia inhibitory factor
needed to start the process of developing an embryonic (LIF). When a blastomere multiplies on such a medium,
stem cell line. By plating it out on an existing cell layer, it the “adapted” daughter cells are isolated and further

TEXTBOX 14.1. The patients involved were not infertile, but had a
Pre-implantation genetic diagnostics. life-threatening disorder in the family. Embryos with
such a congenital anomaly are not implanted in the
Dutch embryo legislation requires that embryos left uterus. The intense political debate that resulted from
over from IVF treatment can be used under certain this authorisation by the Secretary of State, surprised
conditions for medical scientific research. However, the researchers. They thought that embryo selection
they must not be specially created for this purpose. for congenital abnormalities – in this case for the
Through research and debate the Rathenau breast cancer gene – would be a good alternative
Institute 101
stimulates the public to make a judgement to terminating the pregnancy after an amniocentesis
about scientific and technological developments. with an unfavourable outcome for the parents. For the
In 2008 this institute investigated the views of the researchers, abortion seemed morally more difficult
public on the medical scientific use of both left over to justify than embryo selection, for which an embryo
embryos and specially created embryos. This matter had to be created. In the end, the cabinet decided
of embryo use became a sensitive subject later that that the embryo selection could proceed if the existing
year. In mid-2008, after the Secretary of State for multidisciplinary committee in the MUMC+ continued
Public Health had given the Maastricht University to carefully examine each individual case. The
Medical Centre (MUMC+) permission to carry out MUMC+ also examines the seriousness and nature
embryo selection among carriers of a congenital of the disease and the treatment possibilities, and
breast cancer gene, there emerged a serious conflict has to submit new diseases that they want to present
in the cabinet. In the MUMC+ research included for pre-implantation genetic diagnostics (PGD) to a
looking for serious genetic abnormalities in embryos national guidelines committee on PGD (Rathenau
before they were implanted in an IVF treatment. Institute Annual Report 2008).

101
www.rathenau.nl/en.html

Chapter 14: Stem cell therapy: promising and controversial! 259

 cultured as a separate cell line. Human embryonic stem even therapeutic cloning. The genetic material to be
cell lines obtained in this manner have the necessary transferred is usually obtained by performing a skin
pluripotent characteristics, both in vitro and in vivo. biopsy on the patient. The many cells contained in
This second approach uses the remarkable regulatory this piece of skin are allowed to multiply in a culture
capacity of the early mammalian embryo: if one or two medium and are later detached from one another and
cells are missing, it can regenerate the missing cells and from the culture medium using the enzyme trypsin.
form a complete embryo again. The goal is to isolate One of the cells is sucked up in a micropipette and
just one cell of the eight and make a cell line of it. The injected between the cytoplasm and the surrounding
rest of the embryo can then be transplanted directly zona pellucida (protective glycoprotein membrane
into the uterus, since it is still able to implant itself there surrounding the oocyte and the early embryo). The
and generate a complete embryo, fetus and finally a cells are fused using an electrical pulse and the
neonate. It can also be frozen for later transplantation. nucleus containing DNA enters the cytoplasm of the
In fact, this method is no different from the one in which oocyte. After a few more procedures, the resulting
a blastomere is isolated for genetic pre-implantation zygote is grown in vitro in a blastocyst and a human
diagnostics (Textbox 14.1). This approach also evokes stem cell line is developed from this as shown in Figure
the tantalising futuristic scenario in which IVF babies 14.4A. The cells in this line are theoretically genetically
have a genetically compatible embryonic stem cell line identical to the cell from the nucleus and thus identical
in the freezer, which can be used later if necessary for to the cells from the patient, if the donor cell came from
all kinds of stem cell therapies without risk of rejection him or her.
reactions. We certainly haven’t heard the last word on In 2004 and 2005 the first articles claiming success
this. with SCNT were published in leading journals by
The third approach, which until 2006 attracted the South Korean research group led by Hwang.
the most attention in terms of preventing immune However, in 2006, these claims appeared to be
reactions, involves transferring the nucleus of an adult fraudulent. This was a major setback for human
somatic cell, for example a skin cell of the patient to be SCNT and tempered the optimism that had built up
treated, to an oocyte from which the genetic material around it. In mid-2009, SCNT had still not resulted in
has been extracted with a micropipette (Figure14.4C); human embryonic stem cell lines. Scientists continue,
the extracted genetic material concerns one set of untiringly, with their attempts and there have been
chromosomes, because the unfertilised oocyte is a few minor successes (Textbox 14.2). The ethical
haploid (at this stage there is also no question of a real sticking point in this method is that blastocysts made
cell nucleus with a nuclear membrane). This approach in this way can be used not only for therapeutic
is called SCNT or Somatic Cell Nuclear Transfer, or cloning, but also for reproductive cloning. If such a

260 Part 3: Health has limits

TEXTBOX 14.2. that of a normal human embryo and then we’re talking
Human embryos cloned. about more than 5000 genes. However, no stem cell
lines were manufactured from them. ACT did try, but
In the Netherlands and a great many other countries, found that with further cultivating more abnormalities
cloning of people is forbidden. In Belgium, China, Spain appeared. The blastocyst stage was not reached and
and the UK, however, there are some institutes working the standard procedure for generating cell lines (Figure
in this area. In the US, too, cloning is authorised, but 14.4A) could therefore not be used.
receives no federal funding, and it is therefore mainly
FURTHER CULTIVATION OF CLONED
industry that is carrying out research in this particular EMBRYOS REVEALED ABNORMALITIES
domain. Two competing American biotechnology
companies have cloned human embryos with the aim
MMM... THEIR SHAPE
AND COLOR ARE CHANGED!
of developing embryonic stem cell lines. Stemagen, in
California, was first in early 2008. In the journal “Stem
Cells” they wrote that they had made one cloned
human embryo with donor DNA from an adult. With
extensive genetic controls they also demonstrated that
the clone really was a clone. After the Hwang disaster
that was an absolute imperative. Despite the fact that
it is impressive work, it appeared in a low-key journal
and barely caught the world’s attention. A year later
the competitor Advanced Cell Technology (ACT) from The procedure used to make these human clones
Massachusetts published a similar study in “Cloning was the same one used to make Dolly. There were
and Stem Cells”. They had made 19 embryos that more than 200 failed attempts to get to Dolly; at the
survived until they had cleaved three or four times, cost of many ova. The chance of success was, and
i.e. they consisted of 8 or 16 cells. ACT also properly still is, very small. This certainly applies to the cloning
verified that the clones were from an adult human. of donor DNA from adults. Human ova are difficult
During the transition from an undifferentiated embryonic to acquire. In recent years there have been many
stem cell to a final differentiated adult cell, all kinds heated discussions about an alternative way to clone
of genes are deactivated and others just activated. human embryos, for example the use of animal rather
For a successful cloning it must also be possible to than human ova. In mid-2008 the British Houses of
reverse this process. ACT found proof that the gene Parliament voted to legalise these hybrid embryos and
activity of the cloned embryos does indeed resemble as such led the way in this area. However, ACT also

Chapter 14: Stem cell therapy: promising and controversial! 261

 showed in this new study with their genetic analysis “N.Y. to Pay for Eggs for Stem Cell Research.” The
why cloning using this hybrid technique (still) doesn’t article revealed that New York is the first state to allow
work: the genes that were deactivated in the donor researchers funded by state money to pay women to
cell, cannot be reactivated. This explains the failures donate eggs for embryonic stem cell research. Many
to date. scientists are very happy with this decision, but critics
Recently a new controversy looked set to emerge who fear that vulnerable women will be exploited,
on this subject. In The Washington Post of 26 June condemn it outright. What’s certain is that we haven’t
2009 (Anonymous, 2009) there was an article headed heard the last on this subject.

blastocyst were implanted in a uterus, in theory it of such ‘defective’ embryos for the development of
could develop into a full-blown individual, and, what’s cell lines should be easier for some opponents to
more, a 100 percent genetic clone of the DNA donor. accept, because the embryo is not viable as a human
The research is still at the theoretical stage for being, and thus cannot be deprived of an existence
humans, but in mice it was already a reality in 2009 by the creation of cell lines. The zygote made by
(Cyranoski, 2009). Tiny, the first cloned mouse, came nuclear transfer can cleave in vitro and produce cell
about as a result of the reprogramming of a connective masses for the blastocyst, and therefore cell lines, but
tissue cell of her clone parent. And Tiny is no longer the the (induced) genetic defect prevents development
only one. Since Tiny, 27 other cloned mice have been in the uterus. The example given in Figure 14.4D
“born” in this way. One of the males has since created concerns a gene that is essential for the implanting
healthy offspring, born after a normal copulation. The of the embryo in the uterus. Hurlbut is a professor at
“reprogrammed clone premiers” live in Beijing and were Stanford University and has a medical, ethical-medical
created by researchers at the zoological institute of the and theological background. He served for eight years
Chinese Academy of Science. The research report of on the President’s Council on Bioethics. So he’s not
the Chinese scientists was published in Nature (Zhao a newcomer suggesting this dubious alternative102. He
et al., 2009). is at least thinking professionally about these difficult
The fourth approach, the idea for which was launched and ethically controversial matters (Glaser & Hurlbut,
by Hurlbut (Hurlbut, 2005), is in our view based on 2005). Induced pluripotent stem cells (next section)
a dubious premise. The idea starts with a nucleus fortunately may make such an alternative unnecessary.
containing a gene with a mutation brought about by The fifth protocol named in Gruen & Grabel (Gruen &
genetic modification; the pertinent gene is essential Grabel, 2006) avoids the use of human oocytes and
for the embryo to develop in the uterus. The sacrifice embryos to develop genetically compatible human

102
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262 Part 3: Health has limits

embryonic stem cells by fusing a cell from an existing as implausible. By inserting four specific embryonic
human embryonic stem cell line with an adult somatic genes into the genome of specialised mouse cells,
cell (Figure 14.4E). The chromosomes of the original these were reprogrammed into cells that could
embryonic stem cells must then be removed, so that effectively differentiate into any other body cell. In
the cells only have the chromosomes of the somatic other words, specialised cells apparently possess
cell, and thus of the patient. This is necessary for two enough plasticity and can be returned with relatively
reasons. Firstly, the chromosome complement of these straightforward procedures to the pluripotent stage.
hybrids is not stable in time. Secondly, if these cells The disbelief with which the results were received
preserve the DNA of the stem cell line, they are not only spurred on Yamanaka’s research group to
genetically compatible with the patient. The technology generate more convincing proof. This was delivered
to remove all the embryonic stem cell chromosomes ten months later (Okita, Ichisaka, & Yamanaka, 2007)
is not yet available, will probably be very difficult to in an article demonstrating that specialised cells could
produce and is very labour-intensive. In addition, the be reprogrammed into pluripotent stem cells, which
removal has to occur after the reprogramming of the could then differentiate into any specialised body
donor cell DNA, so that the hybrid cell has accepted the cell. To their slightly unpleasant surprise, Yamanaka’s
characteristics of a stem cell. A method for doing that research group had to admit that they were no longer
has, however, not yet been established. Development the first with this proof, since two other laboratories
and testing of this technology will no doubt take years published an article at virtually the same time claiming
and may well be as difficult and expensive as SCNT. that they too had managed this (Maherali et al., 2007;
The technology published in 2007 regarding the Wernig et al., 2007). So began a heated race that
induced reprogramming of adult human cells into was still in full swing during the writing of this chapter
pluripotent cells, places all these approaches in a (mid-2010). In March 2009 alone, four articles were
very different light and probably makes them largely published describing a refinement of the technique.
irrelevant (Baker, 2009). This fierce competitiveness is understandable:
induced pluripotent stem cells are almost as promising
14.7. FORMATION OF INDUCED HUMAN as human embryonic stem cells, but without the ethical
EMBRYONIC PLURIPOTENT STEM CELLS BY objections.
DEDIFFERENTIATION The first embryonic stem cells from mice were isolated
in 1981. It wasn’t until 1998, however, that the same
In August 2006 two Japanese researchers from Kyoto was achieved with human embryonic stem cells. The
University (Takahashi & Yamanaka, 2006) published time between the first induced pluripotent mice stem
a sensational article regarded by many fellow experts cells and those of humans is substantially shorter. By

Chapter 14: Stem cell therapy: promising and controversial! 263

 the end of 2006 pluripotent stem cells (iPS cells) had racing to understand the true nature and utility of the
been induced from mice. By the end of 2007, and early induced pluripotent stem cells (iPS cells).
2008 there were human iPS cells (Park, Zhao et al., Konrad Hochedlinger, a principal faculty member of the
2008; Takahashi et al., 2007; Yu et al., 2007). Induced Harvard Stem Cell Institute, published with colleagues
patient-specific pluripotent stem cells from patients in the April 2010 issue of Nature (Stadtfeld et al., 2010)
with diabetes, Huntington’s disease and muscular that in most mouse iPS cells a cluster of genes, known
dystrophy are described in two articles, which were to be important in development, was not activated. He
published in August 2008 (Dimos et al., 2008; Park, found a small portion of iPS cells in which those genes
Arora et al., 2008); there are still none from embryonic were active, and the cells had the full development
stem cells for people with these diseases. According to potential of embryonic stem cells. Konrad is now
Jeanne Loring, director of the Center for Regenerative repeating the experiment with human cells, and says
Medicine at the Scripps Research Institute in La Jolla, his work suggests that it may be possible to optimise
California, the field of stem cell research is in danger the reprogramming process or to use the genetic
of losing sight of the big questions because of the differences to sort good iPS cells from bad.
competitiveness. Questions such as: what are the In February 2010, researchers from Stanford University
mechanisms of reprogramming and what exactly will School of Medicine published in PLoS ONE that when
reprogrammed cells be able to do on a therapeutic an adult cell is reprogrammed into the embryonic-like
level? She concludes the following: “Making cells is not state, the slate is not wiped clean – cells still have
the end point!” On the contrary, we the authors believe residual gene activity of their original cell type. This
it is only the beginning of the biological challenges - the suggests that for a cell to be completely reset, more
real therapeutic stem cell work. steps might be needed, or certain cell types might be
On 19 July 2010 Carolyn Y. Johnson of the Globe better candidates for reprogramming. Also in February,
Staff writes in The Boston Globe that the breakthrough a study in the journal Stem Cells by researchers of
discovery that scientists could transform adult cells Advanced Cell Technology, a stem cell company in
into stem cells has sparked research in labs across Worcester, Massachusetts, found that blood vessel
the world, spawned start-up companies, and bolstered and retinal cells made from iPS cells aged rapidly. One
the long-term dream that a patient’s own cells could thing is clear, as long as iPS cells have differences,
be used to regenerate damaged tissue. Meanwhile, even slight, their use will be limited, both as potential
scientists have found that these cells, while similar in therapy and as tools to study the origins of disease
many ways to embryonic stem cells, contain subtle or test drugs. It is appropriate to end this section
differences that affect their biology and therapeutic the same as Section 14.3: Time will provide the
potential. Now, researchers all over the world are answers to questions such as whether stem cells are

264 Part 3: Health has limits

ultimately suitable for therapeutic applications and for whether a medical ethics committee is involved to
which disorders, but primarily with which stem cells: protect the rights of patients and whether the proposed
embryonic, adult, or induced pluripotent stem cells, or treatment will be supervised by an official regulatory
perhaps all three? body such as the US FDA. This is an excellent, very
informative and sobering website, including video
14.8. IN CONCLUSION messages from stem cell experts. Visiting this website
is a must for people considering stem cell therapy. The
Stem cell research is in a critical transition phase at the ISSCR has also issued key guidelines for the translation
moment. The first “stem cell products” have reached the of stem cell research into the clinic. These guidelines
clinical test phases and the market is approaching. In are summarised in Textbox 14.3 and come from the
fact an internet search for stem cell therapies results review The bioethics of stem cell research and therapy
in more than 200 companies that claim to grow stem (Hyun, 2010).
cells, inject them back into the patient and cure almost It was a very long time before Geron got permission
any condition (CRC News 1 July 2010). Researchers from the FDA to start the first clinical trials with human
from the Stanford Institute for Stem Cell Biology and embryonic stem cells (Section 14.2). This reflects
Regenerative Medicine warn about these online stem the uncertainty that still surrounds the regulations
cell therapies in the 2 July 2010 issue of the journal Cell on such clinical trials. Questions about the suitability
Stem Cell. In this issue, Dr. Irving Weissman, director of of the regulations in question are increasingly being
this institute, warns of the potential risks to patients and asked, but have until now been obscured by ethical
describes practices and guidelines to assess the validity controversies. Regulations appropriate for these times
of internet claims, such as being wary of clinics that are essential to ensure adequate safety and to gain the
advertise results mainly through patient testimonials. trust of the public, without erecting excessive obstacles
The researchers have launched a website to educate to the development of these products. In 2008 the
and protect patients from unproven stem cell therapies EU led the way with its Advanced Therapy Medicinal
sold online that can be dangerous and very costly. This Products regulation (von Tigerstrom, 2008). On 2
website 103
from the International Society for Stem Cell June 2010 the European Science Foundation (ESF)
Research (ISSCR) includes questions to ask potential released their 38th Science Policy Briefing: Human
clinics, and users can submit a specific website for Stem Cell Research and Regenerative Medicine – A
the society to investigate. When a company or clinic European Perspective on Scientific, Ethical and Legal
is submitted for investigation, the society will evaluate Issues104. In their press release105 of 24 June 2010

103
www.closerlookatstemcells.org//AM/Template.cfm?Section=Home
104
www.esf.org/publications/science-policy-briefings
105
www.esf.org/media-centre/press-releases.html

Chapter 14: Stem cell therapy: promising and controversial! 265

 TEXTBOX 14.3. written consent, and they should demonstrate their
Summary of key ISSCR guidelines for the understanding of the involved risks.
translation of stem cell research into the clinic • Scientists and regulators should work to develop
(Insoo Hyun 2010). common reference standards.
• Appropriate quality standards and management
• Investigators involved in preclinical or clinical research systems for manufacturing cells need to be developed.
involving stem cells or their direct derivatives should • Sufficient preclinical studies in relevant animal models
act within the ISSCR guidelines and other relevant need to be performed.
policies and regulations. • Cells to be used in clinical trials must be extensively
• Clinical research involving stem cells or their direct tested for potential toxicities, including tumorigenicity,
derivatives should be reviewed by human subject in vitro and in animal studies.
review committees supplemented with experts in • Patients should be monitored for long-term health
stem cell science. effects and adverse events reported in a timely
• Donors and patients need to give well-informed manner.

concerning this briefing they summarise the stem-cell • Belgium, Sweden and the UK have adopted
legislation in Europe: legislation to allow the creation of embryos for
research purposes under strict conditions
• Twenty-five countries have adopted legislation • Seventeen countries allow the procurement of stem
which explicitly prohibits human reproductive cells from supernumerary embryos.
cloning (excluding Poland, Lithuania and Ireland as • Bulgaria, Croatia, Cyprus, Luxembourg, Romania
well as Croatia and Luxembourg). and Turkey have not adopted legislation regarding
• Belgium, Sweden, UK, Spain, Finland, the Czech human stem cell research.
Republic and Portugal allow human embryonic
stem cell research and the derivation of new human It is clear that further harmonisation can do no harm:
embryonic stem cell lines from supernumerary (in on the contrary, it is in our opinion a must to clear the
excess) in vitro fertilisation embryos by law. The way for legal, reliable, scientifically proven stem cell
same countries allow somatic cell nuclear transfer therapies!
by law except Finland and the Czech Republic who
neither prohibit nor allow it. N.B. The NIH also has a very informative website on
stem cells106.

106
stemcells.nih.gov/info/basics/basics1.asp

266 Part 3: Health has limits

 14.9. SOURCES Okita, K., Ichisaka, T., & Yamanaka, S. (2007). Generation
of germline-competent induced pluripotent stem cells.
Anonymous. (2009, 26 June). N.Y. to pay for eggs for stem Nature 448, 313-318.
cell research. The Washington Post. Park, I. H., Arora, N., Huo, H., Maherali, N., Ahfeldt, T.,
Baker, M. (2009). Stem cells: Fast and furious. Nature, Shimamura, A., et al. (2008). Disease-specific induced
458(7241), 962-965. pluripotent stem cells. Cell, 134(5), 877-886.
Cyranoski, D. (2009). Mice made from induced stem cells. Park, I. H., Zhao, R., West, J. A., Yabuuchi, A., Huo, H. G.,
Nature, 460(7255), 560-560. Ince, T. A., et al. (2008). Reprogramming of human
Dimos, J. T., Rodolfa, K. T., Niakan, K. K., Weisenthal, L. somatic cells to pluripotency with defined factors.
M., Mitsumoto, H., Chung, W., et al. (2008). Induced Nature, 451(7175), 141-146.
pluripotent stem cells generated from patients with Stadtfeld, M., Apostolou, E., Akutsu, H., Fukuda, A., Follett,
ALS can be differentiated into motor neurons. Science, P., Natesan, S., et al. (2010). Aberrant silencing of
321(5893), 1218-1221. imprinted genes on chromosome 12qF1 in mouse
Glaser, V., & Hurlbut, W. B. (2005). Personal profile - An induced pluripotent stem cells. Nature, 465(7295),
interview with William B. Hurlbut. Rejuvenation 175-181.
Research, 8(2), 110-122. Takahashi, K., Tanabe, K., Ohnuki, M., Narita, M., Ichisaka, T.,
Gruen, L., & Grabel, L. (2006). Concise review: Scientific Tomoda, K., et al. (2007). Induction of pluripotent stem
and ethical roadblocks to human embryonic stem cell cells from adult human fibroblasts by defined factors.
therapy. Stem Cells, 24(10), 2162-2169. Cell, 131(5), 861-872.
Hurlbut, W. B. (2005). Altered nuclear transfer as a morally Takahashi, K., & Yamanaka, S. (2006). Induction of pluripotent
acceptable means for the procurement of human stem cells from mouse embryonic and adult fibroblast
embryonic stem cells. Perspectives in Biology and cultures by defined factors. Cell, 126(4), 663-676.
Medicine, 48(2), 211-228. Varmus, H. (2009). The art and politics of science. New York,
Hyun, I. (2010). The bioethics of stem cell research and WW Norton & Company.
therapy. Journal of Clinical Investigation, 120(1), 71-75. von Tigerstrom, B. J. (2008). The challenges of regulating
Maherali, N., Sridharan, R., Xie, W., Utikal, J., Eminli, S., Arnold, stem cell-based products. Trends in Biotechnology,
K., et al. (2007). Directly reprogrammed fibroblasts show 26(12), 653-658.
global epigenetic remodeling and widespread tissue Wernig, M., Meissner, A., Foreman, R., Brambrink, T.,
contribution. Cell Stem Cell, 1(1), 55-70. Ku, M. C., Hochedlinger, K., et al. (2007). In vitro
Mummery, C., Van de Stolpe, A., & Roelen, B. (2007). reprogramming of fibroblasts into a pluripotent ES-cell-
Stamcellen (2nd edition ed.). Amsterdam, Veen like state. Nature, 448(7151), 318-324.
Magazines. Yu, J., Vodyanik, M. A., Smuga-Otto, K., Antosiewicz-

Chapter 14: Stem cell therapy: promising and controversial! 267

 Bourget, J., Frane, J. L., Tian, S., et al. (2007). Induced Zhao, X. Y., Li, W., Lv, Z., Liu, L., Tong, M., Hai, T., et al.
pluripotent stem cell lines derived from human somatic (2009). iPS cells produce viable mice through tetraploid
cells. Science, 318(5858), 1917-1920. complementation. Nature, 461(7260), 86-90.

268 Part 3: Health has limits

part four
Epilogue
 CASSANDRA

In Greek mythology Cassandra is one of the daughters of Priam, the king of Troy, who lived during the Trojan war.
She was blessed by the god Apollo with the gift of prophecy. The Cassandra syndrome refers these days to an
ominous prediction that later turns out to be true. Will the predictions of the opponents to modern biotechnology
also turn out to be Cassandra prophecies? It certainly seems unlikely now. Compared to other revolutionary
technologies, the calamities caused by modern biotechnology after more than 35 years are non-existent. The
doom scenarios concerning modern biotechnology are very different from those of Cassandra in another way too.
The more Cassandra warned people of an approaching disaster, the less they believed it would happen. In the
figurative sense, Cassandra therefore stands for a prophet of doom, whom nobody believes. That can’t be said
of the opponents of modern biotechnology, who manage to attract attention and support from all possible media.

CASSANDRA

271
15
We started this book by asking
MODERN BIOTECHNOLOGY:
FOR BETTER OR FOR WORSE?

Isn’t biotechnology harmful?

We also said that by the end of the book, we would have drawn the following conclusion:

Biotechnology doesn’t have to be harmful!

We have tried in this book to convince the reader of this proposition. Now that it is finished, we the authors are more
convinced than ever. So we hope that, having read the book, you too can genuinely subscribe to this point of view.
In the writing of this publication we have used literally thousands of articles from websites, newspapers, technical
and scientific journals, books, encyclopaedias, digital newsletters, annual reports, other reports, and so forth. We
have borrowed many sentences, especially from scientific journalists. We have tried to create an anthology of
the many things that have been written in the area of modern biotechnology for the layman. These are frequently
referred to in the text and the more scientific ones also in the list at the end of each chapter. Websites are taken up
as footnotes. To facilitate visiting them, they are also available as a link on the publisher’s website.107 We have spent
years writing what you have just read. However, developments in modern biotechnology move at such high speed
that we have repeatedly had to rewrite parts of the text. We cannot therefore guarantee that references have not
accidentally been omitted, and we offer our apologies should this be the case.
We have restricted ourselves to the subjects that have caused or are still causing controversy. We do, however,
realise that there are many more interesting topics in modern biotechnology, for example, the dawning of the DNA
era in forensic research. We have also made no mention of bio-nanotechnology, bioinformatics, systems biology or
synthetic biology, which are closely related to or follow on naturally from modern biotechnology. All these modern
biotechnologies rely on advanced scientific research and practical entrepreneurship, and their effect on society is
huge.
As with all technologies, the influence of biotechnology on society can be used for good or bad. The decision lies

107
www.wageningenacademic.com/modernbiotech

J. Tramper, Y. Zhu, Modern Biotechnology – Panacea or new Pandora’s box? 273

DOI 10.3920/978-90-8686-725-7_15, © Wageningen Academic Publishers 2011
 with the user of the technology. The biotechnology itself is no more than a means. The biotechnological revolution,
which began in the early ‘70s of the 20th century, will undeniably greatly affect the appearance of the 21st century,
for better or worse.
The scientific journalist Jan Blom expressed a similar view in the final chapter of his book ‘Biotechnologie in
Nederland’ (Biotechnology in the Netherlands), which was published in 1985. Now, 25 years later, this effect is
very noticeable. It is also clear that developments are moving even faster and their impact is even greater than was
initially predicted. Therefore, it is of the utmost importance that modern biotechnology remains a prominent point
of order on society’s agenda, and that this continues to be food for discussion. We must all have a say in deciding
what is permitted, what is not permitted! This new Pandora’s box must be carefully and skilfully opened, so that we
release the gifts and not the curses. We hope that this book has helped.

274 Part 4: Epilogue

 INDEX

GM = genetically modified, also called transgenic in case of GM plants and GM animals

GMO = genetically modified organism

A -recombinant 114

acrylamide 76, 85 -transgenic 31, 40, 123

additives, GM 87, 88, 92, 115, 117 -cloned 43, 123, 124, 125

adeno-associated virus 194 Annan, Kofi 62

adenovirus 193, 194, 197 antibiotic(s)

adjuvant(s) 163, 164 -glycopeptide 157, 162

African Biotechnology Sorghum consortium 62 -resistance 158, 167, 168

allele(s) 201 -semi-synthetic 20, 157, 161, 164, 165

allergy 134-137 anti-rejection drug 211

Alliance for Green Revolution in Africa 62 antisense technology 34

allotransplantation 209 AquAdvantage salmon 41

altitude training 178 artificial organ(s) 224

Amgen 178 Asclepius 151

AMFEP 86, 89 Asilomar conference 26

Amflora potato 63 Aspergillus niger 75, 117

amino acids 21, 76, 105, 134, 237 Association of Manufacturers and

amoxicillin 20, 164, 165 Formulators of Enzyme Products see AMFEB

amphora 97 athlete biological passport 177

ampicillin 164, 165 Augmentin 164

amylase(s) 84, 86, 87

Animal Protection 33, 36, 220 B
animal(s) Baby Fae 211

-ethics 212 baker’s yeast 83

-feed 45, 92, 114, 115, 117 bakery ingredients 93

J. Tramper, Y. Zhu, Modern Biotechnology – Panacea or new Pandora’s box? 277

DOI 10.3920/978-90-8686-725-7, © Wageningen Academic Publishers 2011
 BASF 63, 91, 117 cell(s)
Bacillus thuringiensis 33, 52, 140 -B 214
bacteriophage(s) 166, 167 -bank 257
Barroso, EU Commission President 60, 63 -cross-section 172
Berg, Paul 26, 30 -division 201
Bill and Melinda Gates Foundation 62 -line 251
biological passport, athlete 177 -somatic 247, 258, 260, 266
bio-art exhibition 127 -T 198, 199, 214
bioethics 20 ceftobiprole 160
biofuel(s) 39 cephalosporins 160, 163, 165
biomass-based 39, 50 Chain, Ernst 154
biotechnology, definition 20 champagne 101, 103
blastocyst 251, 252, 256-262 cheese
blastomere 256, 258-260 -genetically modified 79
blood groups 213 -maturation 76
bovine growth hormone, recombinant see rBST -ripening 76
Boyer, Herbert 25, 30 chimeras 208
British Royal Society 143 chromosome 201, 228, 241, 263
bread improvers 81, 84, 86, 93 chymosin 72, 75
Bt Christian Hansen company 72
-resistance 52 cisgenesis 38
-toxin 140 clinical trial(s), phases 157
Bush, George 20, 35, 44, 249, 252 cloning 19, 20, 42, 124, 260-262
Codex Alimentarius guidelines 135, 137
C codon 21
Calgene 34 co-existence 139
carotene, beta 32, 63, 141, 142 Cohen, Stanley 25, 30
Cartagena Protocol 56 Collins, Francis 228, 229, 232-234
Carter, Jimmy 20 complement system 215
casein 72, 78, 122 Confucius 30
Cassandra 271 corn, GM 33, 38, 39, 139
cefalexin 164, 165

278 INDEX
corrective DSM(-Gist) 73, 75, 85, 117, 156, 164
-cell(s) 193, 198 dwarfism 173
-DNA 190
-enzyme(s) 191 E
-gene(s) 190-192 EFSA 45, 54, 136
cotton, GM 33, 38, 140, 146 embryo selection 259
crop(s) environmental safety 52, 54
-Bt 33 enzyme(s)
-H(erbicide) T(olerant) 33 -modified cheese(s) 77
cross-pollination 139 -meat processing 114
Creutzfeldt-Jakob disease 173, 177, 218 epigenetics 243
curd(s), curdling 72, 75, 77 epitope 215
cyclosporin A 211 EPO
-medicine 183
D -performance-enhancing drug 179
Dali, Salvador 239 -test 180
Darwin, Charles Robert 227, 229 EPO, synthesis with
diploid 201 -recombinant animal cells 178
DNA -transgenic mice 182
-analysis 230 -transgenic pigs 182
-corrective 190 erythropoietin see EPO
-hereditary chemical 231 Escherichia coli (E. coli) 25, 31, 174
-two-dimensional structure 21 euchromatic segments 238
-junk 21, 236 EU approval procedure 46
-naked 190 eugenics 200, 234
-passport 36, 232, 234 Eurobarometer 60
-sequencer 231 European Food Safety Authority see EFSA
-zipper 237 exome 240
database, clinical trials gene therapy 192 exon(-skipping) 238
Dolly 19, 41, 253, 261 expression cassette 75
donor shortage 207, 210, 221
dough 83

INDEX 279
 F -technology 20

FDA 34, 35, 85, 103, 123, 249, 265 -therapy 43, 178, 183, 189-205, 232

feeder 259 Generally Recognized As Safe see GRAS

fertility hormone(s) 184, 185 Genetics in a nutshell 201

ficain, ficin 72 genome

Flavr Savr, transgenic tomato 34, 37 -1000-dollar 241

Fleming, Alexander 153-155 -draft version 228

Florey, Howard 154 -sequencing 231

folic acid 63, 163 -surprises 235, 236

follicle-stimulating hormone see FSH genomics 228, 235, 239

food safety 45, 54, 85, 134, 144 genotype 36, 46

food(s), GM 46, 52, 104, 126, 131 Geron 249, 250, 265

Franken(stein)food 69, 124, 131 gluten 83, 89, 91

FSH 184, 185 GMO Compass 47

Fukuyama, Francis 20 GMP 88

Golden Rice 32, 51, 141

G Good Manufacturing Practice see GMP

gelatin, recombinant 120 grape(s), GM 106, 107

Gelsinger, Jesse 193, 194 GRAS 75, 85, 103

Gendicine 197 ‘green’ chemistry 164

gene(s) Green Party 27

-corrective 190 Greenpeace 60, 69, 143

-definition 21 Green Revolution 39, 51, 61, 62

-doping 178, 183, 202-205 growth hormone(s) 41, 118, 173

-dominant 201
-lactase 75 H
-mafia 204 halal 74, 113

-passport 240 haploid 201

-phytase 117 herbicide tolerance 55, 91

-reading 242 Herman, transgenic bull 32, 37, 43

-recessive 201 heterochromatic segments 238

-silencing 90, 145, 201 histone code 242, 243

280 INDEX
histone(s) 241, 242 Kolff, Willem 224
Homer 72 kosher 74, 113
HUGO 235
Human Genome Organisation see HUGO L
human growth hormone labeling 45, 47, 79, 92, 101, 107, 119
-anti-ageing agent 174 lactamase(s), beta 164
-deficiency 173 lactase, gene 75
-performance-enhancing drug 176 lactic acid bacteria 76, 78, 82, 100, 103
Hygieia 151 lactoferrin 32, 39
hyperacute rejection 213-215, 218 LCA 87
legislative framework 57-59
I Life Cycle Assessment see LCA
immobilisation of cells / enzymes 102 lipoprotein-lipase deficiency 195, 196
immune system 32, 194, 214 liposome 190
induced pluripotent stem (iPS) cell(s) see stem cell(s)
inner cell mass 251, 252, 257 M
insect(s) Mad Cow Disease 74, 116, 120, 173
-cell cultures 128 MAdGE 37, 78
-edible 128 maize, GM 55, 59, 62, 90
-resistance 55 malolactic fermentation 100, 103, 104, 110
insulin, human 26, 40 mastitis (udder infection) 32, 118
Intervet 26 meat
intron 238 -artificial 126
IVF 184, 251, 259, 260 -cloned 123, 125
in-vitro fertilisation see IVF -cultivated 126, 127, 128
-happy 128
K -recombinant-free 115
kidney -substitutes 127, 128
-artificial 224 medical revolution(s) 189
-transplant(s) 212, 219 methicillin-resistant Staphylococcus aureus see MRSA
Kluyveromyces lactis 75 mice
Kofi Annan 62 -cloned 262

INDEX 281
 -knock-out 215 phage(s) 166, 167
-Schwarzenegger 202 Pharming 32, 43
milk substitutes 114 pharming 39
modern biotechnology, definition 20 phenotype 46, 201
Monsanto 34, 60, 90, 120 phytase(s) 116, 117
Mothers Against Genetic Engineering see MAdGE phytoremediation 50, 51, 55
MRSA 156, 158 pig(s)
multi-resistant Staphylococcus aureus see MRSA -clones 217, 218
Mummery, Christine 44, 223, 247, 248 -endogenous retrovirus see PERV
muscleman 202 -genome 218, 223
mutation speed 236 -germ-free conditions 212
-knock-out 217, 218
N -multitalented 215
NICE system 78 -spare-part 212
nisin 77, 78 -Sweetie Pie 217
Novozymes 72, 73, 87, 117, 121 -transgenic 211, 215, 217, 218
nucleotide(s) 21 pituitary gland 173, 174, 177
plasmid 31
O plugbug(s) 75
Obama, Barack 35, 249 potato(es), GM 38, 63, 141, 143
Onions, GM 145 Potrykus 51
Prince Charles 35, 126, 133
P prion(s) 120, 173
Panacea 151, 189 precautionary principle 54, 56, 58, 134
pandemic risks 218 processing
Pandemonia 221 -agents 104
Pandora’s box 17, 30, 274 -aids 92
pasteurization 99 profiling method(s) 54
Pasteur, Louis 96, 98 protein synthesis 21, 22
penicillin 153-165 Pruteen 127
Personal Genome Project 240 Pusztai affair 141
PERV 218, 219

282 INDEX
Q RNA 21

Quorn 128 Roundup Ready 33

Roslin Institute 41

R
rapeseed, GM 33, 117 S
rBST 118, 119, 120 Saccharomyces cerevisiae 83, 99, 105

recombinant bovine growth hormone see rBST safety, environmental 52, 54

recombinant DNA (rDNA) technology 31 safety, food 45, 54, 134-136

refuge strategy 140 salmon, GM 41

rejection 207, 211-219, 224, 260 Schneider, Cynthia 24

rejuvenation 175, 210 SCID 193, 197

rennet 72 SCNT 258, 260

rennin 72 serendipity 71

resistance / resistant Severe Combined Immune Deficiency see SCID

-antibiotic 31, 153-165 single cell protein 127

-bacterial strains 156, 157 SMART legislative framework 58

-Bt 52 Somatic Cell Nuclear Transfer see SCNT

-disease 38, 41 sorghum, GM 61, 62, 141, 146

-gene(s) 38, 107 sourdough 82

-herbicides 55, 106 soya, GM 33, 38, 88, 134, 139

-insect(s) 55 splicing 238

-management 39 Staphylococcus aureus 156, 166

-pest 38, 55 starter culture 76, 110

-pesticide 140 stem cell(s)

-plaque 41 -adult 44, 250, 252, 254

retsina 97 -bone marrow 44

ribosomes 22 -embryonic 44, 223, 248

ripening -induced pluripotent 248, 253, 263-265

-agent(s) 77 -line 251, 257, 258

-cheese 76, 77 -multipotent 254, 257

risk analysis 45, 123, 135 -patient-specific 253, 255, 264

risk assessment 54, 135, 138, 219 -piracy 45

INDEX 283
 -pluripotent 248, 254, 256, 257 V
-Presidential list 249, 252 vaccine
-therapy 40, 43, 44, 247 -Immunodrugs 214
-totipotent 256 -swine diarrhoea 26, 31
-unipotent 257 vancomycin 157, 162
steroid(s) 171, 172 Vatican 20, 51, 250
substantial equivalence 46, 136 Venter, J. Craig 24, 228, 231, 234, 239, 241
systems biology 243, 273 vitame A 32, 62, 141
vitamin C 63, 84
T Vitis vinifera 97, 106
teicoplanin 157, 162
tobacco, GM 36, 51, 120 W
tolerance WADA 177, 181, 203, 204
-herbicide 55, 91 Watson, James 26, 239, 241
-stress 91, 106 Wilmut, Ian 41
-value 92 Wine(s)
-zero 93 -definition 96
tomato, GM 38, 51 -GM 108
traceability 45, 92 wheat, GM 59, 82, 89
transcription 22 World Anti-Doping Agency see WADA
translation 22
transgenesis 138 X
transgenic bull Herman 32, 37, 43 xenotransfusion 210
transgenic tomato Flavr Savr 34, 37 xenotransplantation 40, 207-225
transglutaminase 121, 122
trees, GM 52 Y
triplet code 21 yeast(s), GM 102, 104, 120
trophoblast 252, 257
Tropina 127 Z
zipper, DNA 237
U zygote 42, 248, 255, 256, 260, 262
udder infection (mastitis) 32, 118

284 INDEX