D:data/New-age~/Question/Third/III-15.

pm5/Third proof/7-4-05

196 Question Bank of Biochemistry

ORGAN FUNCTION TEST

Q. What are Renal Function Tests? List the Renal function tests? Describe in detail any
one RFT.
Ans. Kidney plays an important role in the maintenance of acid base balance and volume of water
in the body. It serves an important function of excretion of products of metabolism and other
harmful substances. The assessment of renal functions involves urine analysis, blood chemistry,
urography and some special renal function tests.
Each kidney contains about 1 million nephrons. A nephron has a glomerulus and a long tubule that
has three parts
(1) The proximal convoluted tubule (PCT)
(2) The thin loop of Henle (LH)
(3) The distal convoluted tubule (DCT)
The glomeruli are the ultra filter and the filtrate produced is like plasma except that it has almost
no protein, 180 litres of this filtrate is produced in 24 hrs, of which 178 litres of water and most of
the organic and inorganic solutes are reabsorbed. Normally, some components of the filtered solutes
are actively absorbed; glucose, phosphate, amino acids, sodium etc. for some solutes, the maximum
reabsorptive capacity of the tubule is limited and filtered material in excess of this limit is passed on
in the urine. Normal renal threshold for glucose is 180 mg m%; if excess is presented to the
nephrons, it would result in glycosuria. Other solutes are not reabsorbed or are only passively and
partially reabsorbed or are actively secreted by the tubule. Inulin, a carbohydrate used for renal
function studies is not at all reabsorbed by the tubules. Some urea is passively reabsorbed but most
of the filtered urea escapes reabsorption. Exogenous creatinine, H+, K+, phenol red (PSP), para
aminohippurate and penicillin are actively secreted by the tubule cells, thus increasing excretion over
the amount filtered.
Classification of Renal function Tests
On the basis of different functions of kidney the renal function test can be classified as follows
I. Test based on glomerular filtration
Urea Clearance Test
Endogenous Creatinine Clearance Test
Inulin Clearance Test
Cr51 EDTA Test
II. Test based on tubular function
Concentration and Dilution Test
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PSP excretion test

Measurement of tubular secretary mass
III. Tests to measure Renal Plasma Flow (RPF)
Para amino hippurate test (PAH)
Filtration fraction
Maximal Tubular Capacity (Tm)
I. Tests based on Glomerular Filtration
Clearance Tests
Urea clearance, creatinine clearance and inulin clearance test are used to examine for impairment
of glomerular filtration. Clearance is means of expressing quantitatively the rate of excretion of a given
substance by the kidney. This is defined as a volume of blood or plasma, which contains the amount
of substance, which is excreted in the urine in one minute or alternatively, the clearance of a
substance, may be defined as that volume of blood or plasma cleared of the amount of substance
found in one minute excretion of urine.
Urea Clearance Test
The normal blood urea level is in the range of 20 – 40 mg/dl. Uraemia donates a high level of
blood urea and is often due to retention caused by kidney damage. It can also rise in dehydration and
it catabolic states. Urea clearance is defined as the volume of blood or plasma cleared of urea per
minute. If the urine volume exceeds 2 ml/min, the rate of urea elimination is at a maximum and is
directly proportional to the concentration of urea in the blood.
Volume of blood-cleared urea per minute can be calculated from the formula
U×V
B
Where U = concentration of urea in urine (mg).
V = volume of urine in ml/min.
B = the concentration of urea in blood (mg).
Substituting average values, the number of ml of blood cleared of urea per minute
100 × 2.1
= = 75
2.8
It means that the amount of urea excreted in the urine in one minute is equal to the amount found
in 75 ml of blood.
The clearance which occurs when the urinary volume exceeds 2 ml/min is termed as Maximum
Urea Clearance (Cm) and averages normal value is 75.
Cm = 75 ml (normal range 75 ± 10)
Standard Clearance
When the urinary volume is less than 2 ml/min, the rate of urea elimination is reduced, because
relatively more urea is reabsorbed in the tubules, and is proportional to the square root of the urinary
volume. Such clearance is termed as Standard Clearance of Urea (Cs), and average normal value
is 54.
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U× V
Cs = = 54 (normal range 54 ± 10)
B
If a large volume than normal is cleared/mt, renal function is satisfactory. If a smaller volume is
cleared, renal function is impaired.
Expression as %
Sometimes, the result of a urea clearance test is expressed as a % of the normal maximum or of
the normal standard urea clearance depending on whether the urinary output is greater or lesser than
2 ml/min.
Expressed as % of normal
U × V 100
Cm = ×
B 75
% = 1.33
U × V 100
Cs = ×
B 54
% = 1.85
Endogenous Creatinine Clearance Test
At normal levels of creatinine, this metabolite is filtered at the glomerulus, but neither secreted nor
reabsorbed by the tubules. Hence its clearance gives glomerular filtration rate (GFR). This is a
convenient method for estimation of GFR since
(1) It is a normal metabolite in the body.
(2) It does not require the intravenous administration of any test material.
(3) Estimation of creatinine is simple.
Measurement of 24 hour excretion of endogenous creatinine is convenient. This longer collection
period minimizes the timing error.
Inulin Clearance Test
Inulin, a homopolysaccharide, is a polymer of fructose and is ideal substance, as it is not
metabolized in the body. It is excreted entirely through glomerular filtration, being neither excreted nor
reabsorbed by renal tubules. Hence, the number of ml of plasma, which is, cleared of Inulin in one
minute is equivalent to the volume of glomerular filtrate formed in one minute.
Cr51 EDTA Clearance Test
This is simplified single injection method for determination of Cr51 EDTA plasma clearance. This
test is used for routine assessment of Glomerular Filtration Rate (GFR) in adults as well as children.
It is particularly convenient in children where it is not easy to collect 24 hour urine sample. It has
been used for children younger than 1 year old.
Q. What are Liver function Tests. Describe any one test in detail.
Ans. Liver, is a versatile organ, which is involved in metabolism and independently involved in
many other biochemical functions. The regenerating power of liver cells is tremendous. There are
different functions, which are actively performed by liver cells.
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Functions of liver
Metabolic functions
Liver is the key organ and the principle site where the metabolism of carbohydrates, lipids and
proteins take place.
(1) It is the principle organ where cholesterol is synthesized, and catabolized to form bile acids
and bile salts. Esterification of cholesterol solely occurs here.
(2) In this organ the monosaccharides other than glucose are converted to glucose i.e. galactose
is converted to glucose, fructose is converted to glucose.
(3) Liver besides other organs can bring about metabolism of nucleic acids, vitamins and
minerals.
Secretory function
Liver is responsible for the formation and secretion of bile in the intestine. Bile pigments-bilirubin
formed from heme catabolism, is conjugated in liver cells and secreted in the bile.
Excretory function
Certain exogenous dyes like BSP (Bromosulphthalein) and Rose Bengal dye are exclusively
excreted through liver cells.
Detoxication and Protective function
Kupffer cells of liver remove foreign bodies from blood by phagocytosis. Liver cells can
detoxicate drugs, hormones and convert them into less toxic substances for excretion.
Storage function
Liver stores glucose in the form of glycogen. It also stores Vit B12, Vit A etc.
Miscellaneous function
Liver cells are responsible for conversion of preprothrombin (inactive) to active prothrombin in
the presence of Vit K. It also produces other clotting factors like factor V, VII and X. Albumin is
solely synthesized in liver and to some extent α and β globulins also.
Classification of Liver function Tests
The tests used in the study of patients with liver and biliary tract diseases can be classified
according to the specific functions of the liver involved.
I Tests based on abnormalities of pigment metabolism
Serum bilirubin and VD Bergh reaction
Icteric index
Urine bilirubin
Urine and faecal urobilinogen
II Tests based on liver’s role in carbohydrate metabolism
Galactose tolerance test
Fructose tolerance test
Epinephrine tolerance test
III Tests based on changes in plasma proteins
Estimation of total plasma proteins( albumin and globulin).
Flocculation tests
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IV Tests based on Abnormalities of lipids

Cholesterol – Cholesteryl Ester Ratio
V Tests based on the detoxicating function of the liver
Hippuric acid test
VI BSP Tests based on Excretory function of liver
Bromosulphthalein test (– reaction test)
I131 Rose Bengal Test
VII Formation of Prothrombin by liver
Determination of Prothrombin time
VIII Determination of Serum Enzyme activities.
I. Tests based on Abnormalities of Bile pigment metabolism
VD Bergh Reaction and Serum Bilirubin
The methods for detecting and estimating bilirubin in serum are based on the formation of a purple
compound ‘azo-bilirubin’ where bilirubin in serum is allowed to react with a freshly prepared solution
of VD Bergh’s diazo reagent.
Serum bilirubin
It gives a measure of the intensity of jaundice, higher values are found in obstructive jaundice than
in hemolytic jaundice.
Icterix Index
It measures the degree of jaundice by measuring the intensity of the yellow colour of the serum.
The serum or plasma is diluted with physiological saline until it matches in colour a 1 in 10,000
solution of potassium dichromate (Standard solution). The dilution factor is termed the icteric index.
Interpretation
The normal range is from 4 to 6. In latent jaundice, the index is from 7 to 15.
Urine Bilirubin
Most of the tests used for detection of bile pigments depend upon the oxidation of bilirubin to
differently coloured compounds such as biliverdin (green) and bilicyanin (blue).
Interpretation
Bilirubin is found in the urine in obstructive jaundice due to various causes and in ‘cholestasis’.
Conjugated bilirubin can pass through the glomerular filtrate. Bilirubin is not present in most cases of
hemolytic jaundice, as unconjugated bilirubin is carried in plasma attached to albumin hence it cannot
pass through the glomerular filtrate.
Bilirubin in faeces
Bilirubin is not normally present in faeces since bacteria in the intestine reduce it to urobilinogen.
Some may be found if there is very rapid passage of materials among the intestine. It is regularly
found in faces of patients who are being treated with gut sterilizing antibiotics such as neomycin.
Urinary and Faecal Urobilinogen
Urine urobilinogen
Normally there are mere traces of urobilinogen in the urine. Average is 0.64 mg, maximum normal
is 4 mg/24 hours.
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Faecal urobilinogen
Normal quantity of urobilinogen excreted in the faeces per day is from 50 – 250 mg. Since
urobilinogen is formed in the intestine by the reduction of bilirubin the amount of faecal urobilinogen
depends primarily on the amount of bilirubin entering the intestine.
Q. What are Thyroid function tests? Mention any one test in detail.
Ans. The principal hormones secreted by the follicular cells of thyroid are:
Thyroxine (T4)
Tri-iodo thyronine (T3)
‘Reverse’ T3
The hormones T4, T3 and “reverse” T3 are iodinated amino acid tyrosine. The iodine in thyroxine
accounts for 80% of the organically bound iodine in thyroid venous blood. Small amounts of ‘reverse’
tri-iodo thyronine, Monoiodotyrosine (MIT) and other compound are also liberated.
Classification of Thyroid function tests
The assessment of hormones of thyroid gland can be performed by various procedures. But the
most commonly and widely used tests are based on the primary functions of thyroid gland.
Tests based on Primary function of thyroid, viz. Substrate input and hormone synthesis:
Radio-iodine “uptake” studies and ‘turn-over’ (RAI or RIU) studies
PBI131 in serum
T3-suppression test
TSH-stimulation test
TRH-stimulation test
Tests measuring blood levels of thyroid hormones
Serum PBI and BEI levels
Serum T4 levels
Effective Thyroxine ratio(ETR)
Serum T3 levels
Serum TSH level
Erythrocyte uptake of 131 Triodothyronine
Plasma Tyrosine level
Tests based on metabolic effects of thyroid hormones
Serum cholesterol
Serum creatinine
Serum uric acid
Serum CK
BMR
“Scanning” of thyroid gland
Immunological tests to detect auto-immune diseases of thyroid gland
Determination of anti thyroid auto antibodies
Determination of antimicrosomal antibodies