Send Orders for Reprints to reprints@benthamscience.

ae
The Open Bioactive Compounds Journal, 2017, 05, 1-8 1

The Open Bioactive Compounds

Journal
Content list available at: www.benthamopen.com/TOBCJ/

DOI: 10.2174/1874847301705010001

RESEARCH ARTICLE
Cytotoxic Activity of Two Cembranoid Diterpenes from Nicotiana
Sylvestris Against Three Human Cancer Cell Lines
Amir Reza Jassbi1,*, Marzieh Vafapour2, Ardeshir Shokrollahi2, Omidreza Firuzi1, Mehdi Zare1,
Jima N. Chandran3, Bernd Schneider3 and Ian T. Baldwin4
1
Medicinal and Natural Products Chemistry Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
2
Department of Phytochemistry, Yasouj university, Yasouj 75914-353, Iran
3
Research group Biosynthesis/NMR, Max Planck Institute for Chemical Ecology, Jena, Germany
4
Department of Molecular Ecology, Max Planck Institute for Chemical Ecology, Hans-Knöll-Strasse 8, D-07745 Jena,
Germany

Received: May 23, 2017 Revised: August 29, 2017 Accepted: September 01, 2017
Abstract:
Background:
Two cembranoid diterpenes, [(1S, 2E, 4R, 6R, 7E, 11E)-2, 7, 11-cembratriene-4, 6-diol] (1) and its epimer [(1S, 2E, 4S,
6R,7E,11E)-2, 7, 11-cembratriene-4, 6-diol] (2) were isolated from surface dichloromethane washings and chloroform extract of
Nicotiana sylvestris leaves.

Methods:
The compounds were purified using silica gel column- thin layer- and flash column chromatography methods. The structures of the
isolated compounds were elucidated using spectroscopic analysis and their 1H and 13C NMR spectroscopic data were compared with
those of authentic samples reported in the literature. The cytotoxic activity of 1 and 2 against three human cancer cell lines, including
LS180 (human colon adenocarcinoma), MCF-7 (human breast adenocarcinoma) and MOLT-4 (human lymphoblastic leukemia) were
evaluated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) colorimetric bioassay.

Results:
The IC50 values of compounds 1 and 2 were calculated to be between (28.4±3.7 up to 44.0±6.4 µM) (mean±S.E.M.) for the above
mentioned cell lines.

Keywords: Nicotiana sylvestris, Solanaceae, Cembranoid diterpenoids, Anticancer agents, Acyclic diterpenoids, Sesquiterpenoids.

1. INTRODUCTION
Nicotiana sylvestris Spegazzini & Comes is a Solanaceae, known by the common names South American tobacco,
woodland or flowering tobacco. N. sylvestris is a perennial plant, indigenous to northwestern Argentina. This plant is
considered to be one of the ancestors of Nicotiana tabacum [1].
The two cembranoid diterpenes, [(1S,2E,4R,6R,7E,11E)-2,7,11-cembratriene-4,6-diol] (1) and its C-4 epimer,
[(1S,2E,4S,6R,7E,11E)-2,7,11-cembratriene-4,6-diol] (2) together with acyclic diterpenoids, 17-hydroxygeranyllinalool
and two sesquiterpenoids were isolated from the leaves wax of N. sylvestris [2].

* Address correspondence>to this author at the Medicinal and Natural Products Chemistry Research Center, Shahid Abaeiyan Ave, Zip:
71348-53734, Shiraz, Iran, Tel: +98-071-32303872, Fax: +98-071-32332225; E-mails: [email protected], [email protected]

1874-8473/17 2017 Bentham Open

2 The Open Bioactive Compounds Journal, 2017, Volume 05 Jassbi et al.

The cembranoids 1 and its C-4 epimer (2) are two major constituents of different tobacco species [3]. They are
thought to function as insecticides and as inhibitors of plant-growth and fungal-spores germination [4, 5]. Additionally,
they are reported as aldose reductase and prostaglandin inhibitors. They inhibited behavioral sensitization to nicotine in
rats and blocked several types of nicotine acetylcholine receptors [6, 7]. Compounds 1, 2 and their semisynthetic
derivatives, which were transformed by catalytic action of terrestrial and marine bacteria or via chemical
transformation, have been reported as anticancer agents by different authors in animal and in vitro models against
human cancer cells [8 - 11]. However, to the best of our knowledge, their activity against LS180 (human colon
adenocarcinoma), MCF-7 (human breast adenocarcinoma) and MOLT-4 (human lymphoblast leukemia) is discussed
here for the first time.

2. MATERIALS AND METHODS

2.1. General Experimental Procedures

Optical rotations were measured by a Kruss Optronic polarimeter in chloroform. IR spectra were recorded on a
Perkin Elmer Spectrum One FT-IR spectrometer. The 1H and 13C NMR spectra were recorded by Bruker Avance 400
and 500 spectrometer (1H: 400 and 500 MHz, 13C: 100 and 125 MHz) using CDCl3 as solvent and TMS as the internal
standard. Mass spectra were recorded on an Agilent 5975 C inert GC/MSD. TLC analyses were performed on pre
coated silica gel 60 F254 0.5 mm. The TLC plates were impregnated with 5% AgNO3. The silica gel (230-400 mesh)
impregnated with silver nitrate was used as the stationary phase for flash column chromatography (FCC) and silica gel
(70-230 mesh) for the gravity column chromatography (CC).
Fetal bovine serum (FBS), phosphate buffered saline (PBS), RPMI 1640 and trypsin was purchased from Biosera
(Ringmer, UK). Hexane, ethyl acetate, methanol, dichloromethane, dimethyl sulfoxide, silica gel for CC and FCC, the
TLC plates and silver nitrate were purchased from Merck (Darmstadt, Germany). 3-(4,5-dimethylthiazol-2-yl)-2,5-
diphenyltetrazolium bromide (MTT) were obtained from Sigma-Aldrich (St. Louis, MO, USA). Cisplatin was
purchased from EBEWE Pharma (Unterach, Netherlands), penicillin/streptomycin were purchased from Invitrogen (San
Diego, CA, USA).

2.2. Plant Material and Extraction

The seeds of N. sylvestris were obtained from Max Planck Institute for Chemical Ecology (MPICE’s) glass house
and cultivated in the greenhouse of Medicinal and Natural Products Chemistry Research Center (MNCRC) in December
2012. The cultivation procedure was the same as previously reported [12]. Briefly, about 20 seeds after sterilization
were germinated on phytagel agar and kept in a growth chamber with 16/8 h day and night, the day and night
temperature was set at 30 and 20±1°C respectively. After 10 d, the seedlings were transferred to soil and transferred to
glasshouse with the same light and temperature regimes and irrigations once a day. After 34 d, the plants were harvested
and their leaves were subjected to extraction.
At room temperature, the leaves of the plants (136.0 g) were placed in DCM (200 mL) for 30 seconds. Then, the
surface-extracted leaves were subjected to chloroform extraction for three days. The extracts were concentrated in
vacuum at 40°C. The two extracts were subjected to silica gel TLC analyses and after assuring their similarity, were
pooled (2.6 g). AgNO3 (3%)-impregnated silica gel (230-400 mesh, 100 g) was used for purification of the extract’s
constituents. Elution of the column started with n-hexane, continued with stepwise increasing the EtOAc portion up to
100% to increase the polarity of the eluent mixture, and ended up with 5% MeOH in EtOAc. Fractions 18-21, each 75
mL, were checked with 5% AgNO3 impregnated silica gel- TLC and found to be of a similar composition. Similar
fractions were pooled (437 mg). Then flash chromatography was employed (column dimensions: 202 cm, 15 g silica
gel, n-hexane-EtOAc solvent system) to obtain compound 1 (78 mg) as a pure gummy material. Compound 2 (34.0 mg)
was obtained from fraction 22 by flash chromatography (silica gel (230-400 mesh, 5 g), and elution with n-hexane-
EtOAc (1: 1).

2.3. Spectral Data of the Compounds

(1S,2E,4R,6R,7E,11E)-2,7,11-Cembratriene-4,6-diol (1): white powder (78 mg, 0.06% w/w); [α]D + 291.1 (CHCl3);
IR (CHCl3) νmax cm-1: 3347 (OH), 2919 (C-H), 1671 (C=C), 1431, 1373, 1170, (C-O), 1119 (C-C), 1096, 1037, 975, 946,
923, 755, 625. EIMS m/z (rel. int. %): 306 [M]+ C20H34O2 (0.1), 288 [306-H2O] (7), 273 [288-CH3] (6), 260 (14), 245
(21), 230 (4), 217 (13), 203 (10), 189 (15), 189 (6), 176 (19), 163 (18), 149 (11), 136 (32), 121 (44), 109 (58), 108 (51),
Cytotoxic Activity of Two Cembranoid Diterpenes The Open Bioactive Compounds Journal, 2017, Volume 05 3

105 (22), 95 (61), 93 (47), 81 (100), 67 (50), 43 (98) (Fig. 1A). 1H NMR (500 MHz, CDCl3) δ 5.39 (d, J = 15.6 Hz, 1H,
H-3), 5.26 (br. d, J = 9.2 Hz, 1H, H-7), 5.21 (dd, J = 15.6, 9.2 Hz, 1H, H-2), 4.99 (t, J = 5.3 Hz, 1H, H-11), 4.81 (ddd, J
= 9.2, 9.2, 1.4 Hz, 1H, H-6), 2.05 (dd, J = 14.1, 1.4 Hz, 1H, H-5b), 1.92 (dd, J = 12.0, 3.7 Hz, 1H), 1.86 (dd, J = 14.1,
8.9 Hz, 1H, H-5a), 1.70 (d, J = 1.4 Hz, 3H, Me-19), 1.50 (s, 3H, Me-20), 1.39 (s, 3H, Me-18), 0.82 (d, J = 6.7 Hz, 3H,
Me-16), 0.79 (d, J = 6.7 Hz, 3H, Me-17) (Fig. 1C). 13C NMR (125 MHz, CDCl3) δ 136.47 (C-8), 136.10 (C-3), 133.05
(C-12), 131.41 (C-7), 130.40 (C-2), 124.41 (C-11), 71.48 (C-4), 64.45 (C-6), 52.46 (C-5), 46.24 (C-1), 38.84 (C-9),
36.48 (C-13), 32.93 (C-15), 28.73 (C-18), 27.66 (C-14), 23.05 (C-10), 20.60 (C-17), 19.37 (C-16), 15.90 (C-19), 15.00
(C-20) (Fig. 1D).
(1S,2E,4S,6R,7E,11E)-2,7,11-Cembratriene-4,6-diol (2): white powder (34 mg, 0.025% w/w); [α]D: +151.50
(CHCl3); IR (CHCl3) νmax cm-1: 3372 (OH), 2949.3, 1670.0, 1444, 1186, 1123, 1082, 1024, 971, 946.8, 816, 768, 635;
EIMS m/z (rel. int. %): 306 [M+] C20H34O2 (0.98), 288 [306-H2O] (7), 273 [288- CH3] (6), 260 (14), 245 (21), 230 (4),
217 (13), 189 (15), 175 (12), 161 (18), 151 (30), 123 (43), 109 (58), 95 (61), 93 (46), 81 (100), 69 (50), 67 (34), 55 (52),
43 (98) (Fig. 1B). 1H NMR (400 MHz, CDCl3) δ 5.39 – 5.20 (m, 3H, H-2, H-3, H-7), 5.03 (t, J = 5.1 Hz, 1H, H-11),
4.47 (ddd, J = 8.9, 8.9, 2.0 Hz, 1H, H-6), 1.66 (s, 3H, Me-19), 1.51 (s, 3H, Me-20), 1.34 (s, 3H, Me-18), 0.81 (d, J = 6.7
Hz, 3H, Me-16), 0.78 (d, J = 6.7 Hz, 3H, Me-17) Fig. (1E). 13C NMR (100 MHz, CDCl3) δ 137.55 (C-3), 136.80 (C-8),
133.39 (C-12), 130.59 (C-7), 127.79 (C-2), 124.41 (C-11), 72.45 (C-4), 66.34 (C-6), 52.18 (C-5), 46.41 (C-1), 38.86
(C-9), 36.81(C-13), 33.00 (C-15), 30.12 (C-18), 27.95 (C-14), 23.33 (C-10), 20.65 (C-17), 19.34(C-16), 16.07 (C-19),
15.00 (C-20) (Fig. 1F).

2.4. Cell Lines and Culture

The following human cancer cell lines were purchased from the National Cell Bank of Iran, Pasteur Institute,
Tehran, Iran: LS180 (human adenocarcinoma), MCF-7 (human breast adenocarcinoma) and MOLT-4 (human
lymphoblastic leukemia) cells. The cells were cultured in RPMI 1640 medium supplemented with fetal bovine serum
(10% v/v), penicillin (100 units/mL) and streptomycin (100 µg/ml). MOLT-4 cells were grown in suspension, while
LS180 and MCF-7 cells were grown in monolayer cultures in humidified air containing 5% CO2 at 37°C.

2.5. Cytotoxicity Assay

The inhibitory effect of 1 and 2 against cancer cell growth was evaluated by the MTT reduction assay. This
colorimetric assay is based on the conversion of the yellow 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium
bromide (MTT) to the purple formazan by the action of mitochondrial enzyme succinate dehydrogenase in viable cells.
Compounds 1 and 2 were dissolved in dimethylsulfoxide (DMSO) and diluted at least 400 times in growth medium
before being incubated with cells. LS180, MCF-7 and MOLT-4 cells were seeded in 96-well plates at the densities of
50,000, 30,000 and 50,000 cells/mL in 100 µL, respectively and incubated for 24 h. Then, 50 mL of medium was
replaced with fresh medium containing 3-4 different concentrations of the compounds. After 72 h of incubation, the
medium of each well was replaced by fresh RPMI without phenol red containing 0.5 mg/ml MTT and incubated at 37
°C for 4 h. DMSO was used to solubilize the formed formazan crystals. The absorbance of different wells was
measured at 570 nm, with background correction at 655 nm using a microplate reader. The potency of cell growth
inhibition for each compound was expressed as IC50 value, defined as the concentration that caused 50% of maximum
inhibition of cell viability. IC50 values were calculated with software CurveExpert, version 1.3 for Windows.

3. RESULTS AND DISCUSSION

The structures of compounds 1 and 2 were elucidated using spectroscopic analysis, including 1H NMR and APT 13C
NMR together with EIMS spectra and comparing them with those of reference compounds (Fig. 1A-F) [11, 13]. The
stereochemistry of compound 1 was determined by X-ray crystallography (4), and later the absolute configuration of
compound 2 was established by its ozonolysis followed by X-ray crystallography of the resulting degradation product
[14] and at C-6, with chemical transformation to a related cembratriene-4,6,11-triol [15]. The carbon signals in the 13C
NMR spectra of both compounds 1 and 2 had identical or very near chemical shifts to those reported for the authentic
samples in the literature [13]. The 1H NMR spectra of the compounds were compared to those reported for compounds
1 and 2 and their related cembranoids [11, 13].
Inverting the stereochemistry at C-4 affected the chemical shifts of C-4 (δ 71.48 to 72.45), and had about 2 ppm
changes in the chemical shifts at gamma positions: C-2 (δ 130.40 to 127.79) and C-6 (δ 64.45 to 66.34) in compound 1
4 The Open Bioactive Compounds Journal, 2017, Volume 05 Jassbi et al.

compared to those recorded for 2, respectively. The epimerization at C-4 also affected even more drastically the 1H
NMR spectral signals of the olefinic parts of compound 2. In the 1H NMR spectrum of 1, the signals of H-2, H-3 and
H-7 were well resolved and recorded at δ 5.21 (dd, J = 15.6, 9.2 Hz), 5.39 (d, J = 15.6 Hz) and 5.26 (ddd, J = 9.2, 9.2,
1.4 Hz), while in the 1H NMR of 2, these signals are presented as an unresolved multiplet at δ 5.39 – 5.20 due to
overlapping the signals in second-order interactions.
A
A b u n d a n ce

S c a n 1 0 ( 0 . 1 7 9 m in ) : D I R E C T P R O B E _ 9 1 7 . D
43 81
2000000

1800000

1600000
95
121
1400000

1200000
136
1000000

800000 67
149
245
600000 163
108
400000 189 217
203 2 7 32 8 8
200000
176 231 260
30 3 0 63 2 0
0
20 40 60 80 100 120 140 160 180 200 220 240 260 280 300 320
m /z - - >

Fig. 1 contd.....
Cytotoxic Activity of Two Cembranoid Diterpenes The Open Bioactive Compounds Journal, 2017, Volume 05 5

Fig. 1 contd.....
6 The Open Bioactive Compounds Journal, 2017, Volume 05 Jassbi et al.

Fig. (1). EI-MS (A and B), 1H NMR (C and E) and 13C NMR (D and F; 500 MHz in CDCl3) spectra of compounds 1 and 2,
respectively.

The cytotoxic potentials of compounds 1 and 2 were tested against three human cancer cell lines, LS180, MCF-7
and MOLT-4, and were presented as IC50s in Table (1) in comparison to the standard anticancer agent cisplatin. The
inhibitory effect of compounds 1 and 2 against tumor promotion has been previously reported. It has been shown that
Cytotoxic Activity of Two Cembranoid Diterpenes The Open Bioactive Compounds Journal, 2017, Volume 05 7

these compounds are able to inhibit the skin tumor promoting effect of 12-O-tetradecanoylphorbol-13-acetate (TPA) in
mice [8]. The tumorigenisis inhibition of cembranoid diterepenoids against prostate cancer cell lines has also been
reported for different derivatives of cembranoids (1S,2E,4S,6E,8S,11E)-2,6,11-cembratriene-8-O-methyl-4,8-diol and
the known (1S,2E,4R,6R,7E,11E)-2,7,11-cembratriene-4-O-methyl-4,6-diol isolated from N. tabacum and their
biotransformed, mostly oxidized products, by the action of two fungal strains: Cunninghamella NRRL 5695 and Mucor
ramannianus ATCC 9628 [9]. Compound 1 was converted into its C-6 hydroxyl esters with different carbamate
functionalities and also to different hydroxylation reaction biocatalyzed by symbiotic Bacillus species isolated from a
Red sea marine sponge [11]. Compound 1 and its derivatives were tested on highly invasive prostate cancer cell lines
and a SAR conclusion indicated their high potential against cancer cell lines due to the polar substitution on C-6
hydroxy group [11].
The semisynthetic products and a secocembranoid diterpenoid resulted from the reaction of compound 1 with
different halogenated carbamic acids and also reactions catalyzed in the presence of marine Bacillus species and M.
ramannianus ATCC 9628 and C. elegans showed antiproliferative activity against highly malignant +SA mammary
epithelial cells with an IC50 range of 15–30 μM [10].
The relatively good cytotoxic activity of compounds 1 and 2 (Table 1) in comparison to the standard anticancer
agent cisplatin against the tested cancer cell lines in the present study is compatible with the previous reports.
Table 1. Cytotoxic activity of compounds 1 and 2 against human cancer lines determined by MTT reduction assay.

IC50 (μM)
Sample
LS-180 MCF-7 MOLT-4
Compound 1 42.6 ± 3.2 44.0 ± 6.4 39.4 ± 4.8
Compound 2 42.8 ± 2.1 35.2 ± 3.6 28.4 ± 3.7
Cisplatin 5.9 ± 2.4 12.6 ± 5.8 3.1 ± 0.8
The results are presented as mean ± S.E.M. of 3-4 experiments.

ETHICS APPROVAL AND CONSENT TO PARTICIPATE

Not applicable.

CONSENT FOR PUBLICATION

Not applicable.

CONFLICT OF INTEREST
The authors declare no conflict of interest, financial or otherwise.

ACKNOWLEDGEMENTS
The authors are thankful to financial supports of Shiraz University of Medical Sciences, the University of Yasooj,
Iran, and Alexander von Humboldt Foundation (grant number: 3.4 - IRN/1101775).
REFERENCES

[1] Gray JC, Kung SD, Wildman SG, Sheen SJ. Origin of Nicotiana tabacum L detected by polypeptide composition of Fraction I protein. Nature
1974; 252(5480): 226-7.
[http://dx.doi.org/10.1038/252226a0] [PMID: 4421263]
[2] Wallin I, Narbonne C, Wahlberg I, Nishida T, Enzell CR. Two new acyclic diterpenoids from Nicotiana sylvestris. Acta Chem Scand B 1980;
34: 391-6.
[http://dx.doi.org/10.3891/acta.chem.scand.34b-0391]
[3] Roberts DL, Rowland RL. Macrocyclic Diterpenes. α- and β-4,8,13-Duvatriene-1,3-diols from Tobacco1. J Org Chem 1962; 27: 3989-95.
[http://dx.doi.org/10.1021/jo01058a056]
[4] Springer JP, Clardy J, Cox RH, Cutler HG, Cole RJ. The structure of a new type of plant growth inhibitor extracted from immature tobacco
leaves. Tetrahedron Lett 1975; 16: 2737-40.
[http://dx.doi.org/10.1016/S0040-4039(00)75227-0]
[5] Wahlberg I, Enzell C. Tobacco cembranoids. Beitr Tabakforsch Int 1984; 12: 93-104.
[http://dx.doi.org/10.2478/cttr-2013-0530]
[6] Olsson E, Holth A, Kumlin E, Bohlin L, Wahlberg I. Structure-related inhibiting activity of some tobacco cembranoids on the prostaglandin
8 The Open Bioactive Compounds Journal, 2017, Volume 05 Jassbi et al.

synthesis in vitro. Planta Med 1993; 59(4): 293-5.

[http://dx.doi.org/10.1055/s-2006-959684] [PMID: 8372141]
[7] Ferchmin PA, Lukas RJ, Hann RM, et al. Tobacco cembranoids block behavioral sensitization to nicotine and inhibit neuronal acetylcholine
receptor function. J Neurosci Res 2001; 64(1): 18-25.
[http://dx.doi.org/10.1002/jnr.1049] [PMID: 11276047]
[8] Saito Y, Takizawa H, Konishi S, Yoshida D, Mizusaki S. Identification of cembratriene-4,6-diol as antitumor-promoting agent from cigarette
smoke condensate. Carcinogenesis 1985; 6(8): 1189-94.
[http://dx.doi.org/10.1093/carcin/6.8.1189] [PMID: 2990757]
[9] Baraka HN, Khanfar MA, Williams JC, El-Giar EM, El Sayed KA. Bioactive natural, biocatalytic, and semisynthetic tobacco cembranoids.
Planta Med 2011; 77(5): 467-76.
[http://dx.doi.org/10.1055/s-0030-1250478] [PMID: 21049399]
[10] El Sayed KA, Laphookhieo S, Yousaf M, et al. Semisynthetic and biotransformation studies of (1S,2E,4S,6R,7E,11E)-2,7,11-
cembratriene-4,6-diol. J Nat Prod 2008; 71(1): 117-22.
[http://dx.doi.org/10.1021/np0704351] [PMID: 18177013]
[11] El Sayed KA, Laphookhieo S, Baraka HN, et al. Biocatalytic and semisynthetic optimization of the anti-invasive tobacco
(1S,2E,4R,6R,7E,11E)-2,7, 11-cembratriene-4,6-diol. Bioorg Med Chem 2008; 16(6): 2886-93.
[http://dx.doi.org/10.1016/j.bmc.2007.12.056] [PMID: 18222089]
[12] Jassbi AR, Gase K, Hettenhausen C, Schmidt A, Baldwin IT. Silencing geranylgeranyl diphosphate synthase in Nicotiana attenuata
dramatically impairs resistance to tobacco hornworm. Plant Physiol 2008; 146(3): 974-86.
[http://dx.doi.org/10.1104/pp.107.108811] [PMID: 17965175]
[13] Wahlberg I, Arndt R, Wallin I, Vogt C, Nishida T, Enzell CR. Tobacco Chemistry. 59. Six New Cembratrienetriols from Tobacco. Acta Chem
Scand B 1984; 38: 21-30.
[http://dx.doi.org/10.3891/acta.chem.scand.38b-0021]
[14] Aasen AJ, Junker N, Enzell CR, Berg J-E, Pilotti A-M. Tobacco chemistry 36. Absolute configuration of tobacco thunberganoids.
Tetrahedron Lett 1975; 16: 2607-10.
[http://dx.doi.org/10.1016/S0040-4039(00)75193-8]
[15] Wahlberg I, Wallin I, Narbonne C, Nishida T, Enzell CR, Berg J. Tobacco chemistry. 55. Three new cembranoids from Greek tobacco. The
stereochemistry of (1S, 2E, 4S, 6R, 7E, 11E)-2, 7, 11-cembratriene-4, 6-diol. Acta Chem Scand B 1982; 36: 3.

© 2017 Jassbi et al.

This is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International Public License (CC-BY 4.0), a
copy of which is available at: https://creativecommons.org/licenses/by/4.0/legalcode. This license permits unrestricted use, distribution, and
reproduction in any medium, provided the original author and source are credited.