Editorial 2 2 I have recently sent out a letter to many of our colleagues and friends in the Iranian scientific community informing them of the recent addition of AJMB to two of the world’s largest databases, namely: Scopus and Embase. In case the spectrum of the coverage by these databases are not known to the AJMB readers in Iran, I have decided to provide you with some information about these two databases obtained from the official company website. Scopus (launched in November 2004) is a database of abstracts and citations and covers nearly 18,000 titles from more than 5,000 international publishers, including coverage of 16,500 peer-reviewed journals in the scientific, technical, medical and social sciences as well as fields in arts and humanities. It is owned by Elsevier and searches in Scopus incorporate searches of scientific web pages (435 Million), patent databases (23 Million) from 5 patent offices (US Patent and Trademark Office, European Patent Office, Japan Patent Office, World Intellectual Property Organization and UK Intellectual Property Office), article-in-press from over 3000 journals and full coverage of Medline titles. Embase or the Excerpta Medica Database is a biomedical and pharmacological database produced also by Elsevier and contains over 11 million records from 1947 to the present date. Each record is fully indexed and covers over 5,000 biomedical journals from 70 countries and is available online through a number of database vendors. Embase has a holding of more than 2,000 biomedical titles that are not offered by the Medline. Embase delivers comprehensive, authoritative, and reliable coverage of the most relevant biomedical literature. Now that AJMB has been recognized as a journal deserving to be included in these databases, it is up to the Iranian scientists to support the AJMB’s mission of publishing high quality articles in the field of Medical Biotechnology from Iran and worldwide. I continue to look forward to receiving your high quality articles and would appreciate if you could inform and encourage your colleagues to submit their articles to AJMB and enjoy its international exposure among thousands of other medical scientific journals in the world. 68 68 Ali M. Ardekani Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Editorial 158.pdf ####

Production of Monoclonal Antibody against Human Nestin 2 2 We have employed a peptide-based antibody generation protocol for producing antibody against human nestin. Using a 12-mer synthetic peptide from repetitive region of human nestin protein devoid of any N- or O-glyco-sylation sequences, we generated a mouse monoclonal antibody capable of recognizing human, mouse, bovine, and rat nestin. A wide variety of nestin proteins ranging from 140-250 kDa was detected by this antibody. This antibody is highly specific and functional in applications such as ELISA, flow cytometry, immunocytochemistry, and Western blot assays. 69 77 Reza Hadavi Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Iran Amir-Hassan Zarnani Nanobiotechnology Research Center, Avicenna Research Institute, ACECR Nanobiotechnology Research Center, Avicenna Research Institute, ACECR Iran Negah Ahmadvand Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Iran Ahmad Reza Mahmoudi Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Iran Ali Ahmad Bayat Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Iran Jafar Mahmoudian Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Iran Mohammad Reza Sadeghi Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Haleh Soltanghoraee Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Mohammad Mehdi Akhondi Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Majid Tarahomi Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Mahmood Jeddi-Tehrani Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Iran Hodjattallah Rabbani Monoclonal Antibody Research Center, Avicenna Research Institute, ACECRImmune and Gene Therapy Lab, CCK, Department of Oncology-Pathology, Karolinska University Hospital Solna, Karolinska Institutet Monoclonal Antibody Research Center, Avicenna Research Institute, ACECRImmune and Gene Therapy Lab, CCK, Department of Oncology-Pathology, Karolinska University Hospital Solna, Karolinska Institutet IranSweden AntibodiesBlottingImmunohistochemistryMonoclonalNestinPeptidesWestern 32.pdf Lendahl U, Zimmerman LB, McKay RD. CNS stem cells express a new class of intermediate filament protein. Cell 1990;60(4):585-595. ##Amoh Y, Li L, Katsuoka K, Hoffman RM. Multipotent nestin-expressing hair follicle stem cells. J Dermatol 2009;36(1):1-9. ##Kawamoto M, Ishiwata T, Cho K, Uchida E, Korc M, Naito Z, et al. Nestin expression correlates with nerve and retroperitoneal tissue invasion in pancreatic cancer. Hum Pathol 2009;40(2):189-198. ##Leach SD. Epithelial differentiation in pancreatic development and neoplasia: new niches for nestin and Notch. J Clin Gastroenterol 2005;39(4 Suppl 2):S78-S82. ##Rogelsperger O, Ekmekcioglu C, Jager W, Klimpfinger M, Konigsberg R, Krenbek D, et al. Coexpression of the melatonin receptor 1 and nestin in human breast cancer specimens. J Pineal Res 2009; 46(4):422-432. ##Loja T, Chlapek P, Kuglik P, Pesakova M, Oltova A, Cejpek P, et al. Characterization of a GM7 glioblastoma cell line showing CD133 positivity and both cytoplasmic and nuclear localization of nestin. Oncol Rep 2009;21(1):119-127. ##Rushing EJ, Sandberg GD, Horkayne-Szakaly I. High-Grade astrocytomas show increased Nestin and Wilms's tumor gene (WT1) protein expression. Int J Surg Pathol 2009. [Epub ahead of print] ##Sellheyer K, Nelson P, Krahl D. Dermatofibrosarcoma protuberans: a tumour of nestin-positive cutaneous mesenchymal stem cells? Br J Dermatol 2009;161(6):1317-1322. ##Yamada H, Takano T, Ito Y, Matsuzuka F, Miya A, Kobayashi K, et al. Expression of nestin mRNA is a differentiation marker in thyroid tumors. Cancer Lett 2009;280(1):61-64. ##Messam CA, Hou J, Major EO. Coexpression of nestin in neural and glial cells in the developing human CNS defined by a human-specific anti-nestin antibody. Exp Neurol 2000;161(2):585-596. ##Kohler G, Milstein C. Continuous cultures of fused cells secreting antibody of predefined specificity. Nature 1975;256(5517):495-497. ##Shojaeian S, Allameh A, Zarnani AH, Chamankhah M, Ghods R, Bayat AA, et al. Production and characterization of monoclonal antibodies against the extracellular domain of CA 125. Immunol Invest 2010;39(2):114-131. ##Grigelioniene G, Blennow M, Torok C, Fried G, Dahlin I, Lendahl U, et al. Cerebrospinal fluid of newborn infants contains a deglycosylated form of the intermediate filament nestin. Pediatr Res 1996; 40(6):809-814. ##Kleeberger W, Bova GS, Nielsen ME, Herawi M, Chuang AY, Epstein JI, et al. Roles for the stem cell associated intermediate filament Nestin in prostate cancer migration and metastasis. Cancer Res 2007;67(19):9199-9206. ##Jin ZG, Liu L, Zhong H, Zhang KJ, Chen YF, Bian W, et al. Second intron of mouse nestin gene directs its expression in pluripotent embryonic carcinoma cells through POU factor binding site. Acta Biochim Biophys Sin 2006;38(3):207-212. ##Corsino PE, Davis BJ, Norgaard PH, Parker NN, Law M, Dunn W, et al. Mammary tumors initiated by constitutive Cdk2 activation contain an invasive basal-like component. Neoplasia 2008;10(11): 1240-1252. ##Gil YG, Kang MK. Capsaicin induces apoptosis and terminal differentiation in human glioma A172 cells. Life Sci 2008;82(19-20):997-1003. ##Veselska R, Kuglik P, Cejpek P, Svachova H, Neradil J, Loja T, et al. Nestin expression in the cell lines derived from glioblastoma multiforme. BMC Cancer 2006;6:32. ##Wu A, Oh S, Wiesner SM, Ericson K, Chen L, Hall WA, et al. Persistence of CD133+ cells in human and mouse glioma cell lines: detailed characterization of GL261 glioma cells with cancer stem cell-like properties. Stem Cells Dev 2008;17(1): 173-184. ##Rappa G, Mercapide J, Anzanello F, Prasmickaite L, Xi Y, Ju J, et al. Growth of cancer cell lines under stem cell-like conditions has the potential to unveil therapeutic targets. Exp Cell Res 2008;314 (10):2110-2122. ##

Angiotensin II Differentially Induces Matrix Metalloproteinase-9 and Tissue Inhibitor of Metalloproteinase-1 Production and Disturbs MMP/TIMP Balance 2 2 Angiotensin II, the main component of the renin-angiotensin system, is associated with cardiovascular diseases such as hypertension, vascular remodeling and inflammation. Remodeling process results from dysregulation of Matrix Metalloproteinases (MMPs) and their tissue inhibitors (TIMPs). MMPs are considered as important target genes for angiotensin II. The aim of this study was to determine the effects of angiotensin II on MMP-9 and TIMP-1 production and MMP/TIMP balance in a monocytic cell type. Human monocytic U-937 cells were cultured and treated with 100 nM angiotensin II. Supernatants were analyzed for MMP-9 and TIMP-1 using ELISA and zymography methods. Real-time PCR was utilized to evaluate relative MMP-9 and TIMP-1 genes expression following treatments. Cytotoxicity potentials of treatments were determined by assaying lactate dehydrogenase leakage from the cells. Stimulation of the monocytic cells with angiotensin II significantly increased MMP-9 and TIMP-1 secretion as measured by ELISA (p<0.05). It also augmented gelatinolytic activity of MMP-9 in the conditioned media as much as 49% (p<0.05). Incubation of the cells with angiotensin II for 12 hr increased MMP-9 and TIMP-1 gene expression 2.7 and 1.8 folds, respectively (p<0.05). Angiotensin II treatments did not establish significant cytotoxic effects. In summary, our data provide further evidences that monocytic MMP-9 is a major effector of angiotensin II. It is induced more efficiently than TIMP-1 by angiotensin II that leads to MMP/TIMP imbalance. Our data also reveal the pivotal participation of these cells in pathological cardiovascular remodeling mediated by angiotensin II. 79 86 Hamid Yaghooti Department of Biochemistry, School of Medicine, Iran University of Medical Sciences Department of Biochemistry, School of Medicine, Iran University of Medical Sciences Iran Mohsen Firoozrai School of Medicine, Iran University of Medical Sciences, Tehran, Iran School of Medicine, Iran University of Medical Sciences, Tehran, Iran Iran Soudabeh Fallah School of Medicine, Iran University of Medical Sciences School of Medicine, Iran University of Medical Sciences Iran Mohammad Reza Khorramizadeh Department of Medical Biotechnology, School of Advanced Medical Technologies, Tehran University of Medical Sciences Department of Medical Biotechnology, School of Advanced Medical Technologies, Tehran University of Medical Sciences Iran Angiotensin IIMatrix metalloproteinase 9Monocytic cellTissue inhibitor of metalloproteinase-1 33.pdf Mehta PK, Griendling KK. Angiotensin II cell signaling: physiological and pathological effects in the cardiovascular system. Am J Physiol Cell Physiol 2007;292:82-97. ##Unger T. The role of the renin-angiotensin system in the development of cardiovascular disease. Am J Cardiol 2002;89(2):3A-9A. ##Cheng ZJ, Vapaatalo H, Mervaala E. Angiotensin II and vascular inflammation. Med Sci Monit 2005; 11(6):RA194-205. ##Galis ZS, Khatri JJ. Matrix metalloproteinases in vascular remodeling and atherogenesis: the good, the bad, and the ugly. Circ Res 2002;90:251-262. ##Visse R, Nagase H. Matrix metalloproteinases and tissue inhibitor of metalloproteinases. Circ Res 2003;92:827-839. ##Raffetto JD, Khalil RA. Matrix metalloproteinases and their inhibitors in vascular remodeling and vascular disease. Biochem Pharmacol 2008;75(2): 346-359. ##Li YY, Feldman AM, Sun Y, McTiernan CF. Differential expression of tissue inhibitors of metalloproteinases in the failing human heart. Circulation 1998;98:1728-1734. ##Browatzki M, Larsen D, Pfeiffer CAH, Gehrke SG, Schmidt S, Kranzhofer A, et al. Angiotensin II stimulates matrix metalloproteinase secretion in human vascular smooth muscle cells via nuclear factor-?B and activator protein-1 in a redox sensitive manner. J Vasc Res 2005;42(5):415-423. ##Wang TL, Yang YH, Chang H, Hung CR. Angiotensin II signals mechanical stretch-induced cardiac matrix metalloproteinase expression via JAK-STAT pathway. J Mol Cell Cardiol 2004;37(3):785-794. ##Luchtefeld M, Grote K, Grothusen C, Bley S, Bandlow N, Selle T, et al. Angiotensin II induces MMP-2 in a p45phox-dependent manner. Biochem Biophys Res Commun 2005;328(1):183-188. ##Kim MP, Zhou M, Wahl LM. Angiotensin II increases human monocyte matrix metalloproteinase-1 through the AT2 receptor and prostaglandin E2: implications for atherosclerotic plaque rupture. J Leukoc Biol 2005;78:195-201. ##Kanome T, Watanabe T, Nishio K, Takahashi K, Hongo S, Miyazaki A. Angiotensin II upregulates acyl- CoA: cholesterol acyltransferase-1 via the angiotensin II type 1 receptor in human monocytemacrophages. Hypertens Res 2008;31(9):1801-1810. ##Rezaei A, Ardestani SK, Forouzandeh M, Tavangar SM, Khorramizadeh MR, Payabvash S, et al. The effects of N-acetylcysteine on the expression of matrix metalloproteinase-2 and tissue inhibitor of matrix metalloproteinase-2 in hepatic fibrosis in bile duct ligated rats. Hepatol Res 2008;38(12): 1252-1263. ##Fielitz J, Leuschner M, Zurbrugg HR, Hannack B, Pregla R, Hetzer R, et al. Regulation of matrix metalloproteinases and their inhibitors in the left ventricular myocardium of patients with aortic stenosis. J Mol Med 2004;82(12):809-820. ##Yang D, Ma S, Li D, Tang B, Yang Y. Angiotensin II receptor blockade improves matrix metalloproteinases/ tissue inhibitor of matrix metalloproteinase-1 balance and restores fibronectin expression in rat infarcted myocardium. Biochem Biophys Res Commun 2009;388(3):606-611. ##Shah PK, Falk E, Badimon JJ, Fernandez-Ortiz A, Mailhac A, Villareal-Levy G. Human monocyte-derived macrophages induce collagen breakdown in fibrous caps of atherosclerotic plaques. Potential role of matrix-degrading metalloproteinases and implications for plaque rupture. Circulation 1995; 92(6):1565-1569. ##Fotheringham JA, Mayne MB, Grant JA, Geiger JD. Activation of adenosine receptors inhibits tumor necrosis factor–alpha release by decreasing TNF-alpha mRNA stability and p38 activity. Eur J Pharmacol 2004;497(1):87-95. ##Yuan ZY, Nimata M, Okabe T, Shioji K, Hasegawa K, Kita T, et al. Olmesartan, a novel AT(1) antagonist, suppresses cytotoxic myocardial injury in autoimmune heart failure. Am J Physiol Heart Circ Physiol 2005;289:H1147-1152. ##Vincenti MP, Brinckerhoff CE. Signal transduction and cell-type specific regulation of matrix metalloproteinase gene expression: can MMPs be good for you? J Cell Physiol 2007;213:355-364. ##Fiotti N, Altamura N, Fisicaro M, Carraro N, Uxa L, Grassi G, et al. MMP-9 microsatellite polymorphism and susceptibility to carotid arteries atherosclerosis. Arterioscler Thromb Vasc Biol 2006;26:1330-1336. ##Castoldi G, di Gioia CRT, Travaglini C, Busca G, S Redaelli, Bombardi C, et al. Angiotensin II increases tissue-specific inhibitor of metalloproteinase-2 expression in rat aortic smooth muscle cells in vivo: evidence of a pressure-independent effect. Clin Experi Pharmcol Physiol 2007;34(3):205-209. ##Chua CC, Hamdy RC, Chua BHL. Angiotensin II induces TIMP-1 production in rat heart endothelial cells. BBA-Mol Cell Res 1996;1311(3):175-180. ##Zhang W, Zhong M, Yang G, Li JP, Guo C, Wang Z, et al. Matrix metalloproteinase-9/tissue inhibitors of metalloproteinase-1 expression and atrial structural remodeling in a dog model of atrial fibrillation: inhibition with angiotensin-converting en-zyme. Cardiovasc Pathol 2008;17(6):399-409. ##Wang XD, Chen XM, Wang JZ, Hong Q, Feng Z, Fu B, et al. Signal transducers and activators of transcription 3 mediates up-regulation of angiotensin II-induced tissue inhibitor of metalloproteinase-1 expression in cultured human senescent fibroblasts. Chin Med J 2006;119(13):1094-1102. ##Yang BC, Phillips MI, Mohuczy D, Meng H, Shen L, Mehta P, et al. Increased angiotensin II type 1 receptor expression in hypercholesterolemic atherosclerosis in rabbits. Arterioscler Thromb Vasc Biol 1998;18:1433-1439. ##

Conjugation of R-Phycoerythrin to a Polyclonal Antibody and F (ab')2 Fragment of a Polyclonal Antibody by Two Different Methods 2 2 R-Phycoerythrin (R-PE), a fluorescent protein from phycobiliprotein family, is isolated from red algae. Conjugation of antibodies to R-PE facilitates multiple fluorescent staining methods. In the present study polyclonal antibodies and polyclonal F(ab')2 fragment antibodies were conjugated to R-PE by two different methods. The efficiency of the methods was evaluated using Immunocytochemistry (ICC) and Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE). In the first conjugation method, PE was attached to SMCC linker followed by conjugation of antibody to PE-SMCC. In the second method, SH groups were added onto R-PE molecule, while the antibody was attached to SPDP linker. Then, the antibody-SPDP molecule was conjugated to R-PE. Our results showed that the two conjugation methods did not have any abrogative effects on the antibody binding activity. 87 92 Jafar Mahmoudian Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Iran Mahmood Jeddi-Tehrani Monoclonal Antibody Research Center, Avicenna Research Institute, ACECRImmune and Gene Therapy Lab, Cancer Center Karolinska, Karolinska Institute Monoclonal Antibody Research Center, Avicenna Research Institute, ACECRImmune and Gene Therapy Lab, Cancer Center Karolinska, Karolinska Institute IranSweden Hodjattallah Rabbani Monoclonal Antibody Research Center, Avicenna Research Institute, ACECRImmune and Gene Therapy Lab, Cancer Center Karolinska, Karolinska Institute Monoclonal Antibody Research Center, Avicenna Research Institute, ACECRImmune and Gene Therapy Lab, Cancer Center Karolinska, Karolinska Institute IranSweden Ahmad Reza Mahmoudi Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Iran Mohammad Mehdi Akhondi Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Amir-Hassan Zarnani Nanobiotechnology Research Center, Avicenna Research Institute, ACECR Nanobiotechnology Research Center, Avicenna Research Institute, ACECR Iran Leila Balaei Goli Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Iran Mahdokht Babaei Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Iran Roya Ghods Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Iran AntibodyConjugationImmunocytochemistryPhycoerythrin 34.pdf Glazer AN. Phycobilisomes: structure and dynamics. Annu Rev Microbiol 1982;36(1):173-198. ##Jiang T, Zhang J, Chang W, Liang D. Crystal structure of R-phycocyanin and possible energy transfer pathways in the phycobilisome. Biophys J 2001;81(2):1171-1179. ##Hardy RR. Purification and coupling of fluorescent proteins for use in flow cytometry. Handbook of experimental immunology. 4th ed. Oxford: Black-well Scientific; 1986. ##Chambers JD, Simon SI, Berger EM, Sklar LA, Arfors KE. Endocytosis of beta 2 integrins by stimulated human neutrophils analyzed by flow cytometry. J Leukoc Biol 1993;53(4):462-469. ##Kronick MN, Grossman PD. Immunoassay techniques with fluorescent phycobiliprotein conjugates. Clin Chem 1983;29(9):1582-1586. ##Oi VT, Glazer AN, Stryer L. Fluorescent phycobiliprotein conjugates for analyses of cells and molecules. J Cell Biol 1982;93(3):981-986. ##Ferrante A, Thong YH. Optimal conditions for simultaneous purification of mononuclear and polymorphonuclear leucocytes from human blood by the hypaque-ficoll method. J Immunol Methods 1980;36(2):109-117. ##Migneault I, Dartiguenave C, Bertrand MJ, Waldron KC. Glutaraldehyde: behavior in aqueous solution, reaction with proteins, and application to enzyme crosslinking. BioTechniques 2004;37(5): 790-802. ##Mattson G, Conklin E, Desai S, Nielander G, Savage MD, Morgensen S. A practical approach to crosslinking. Mol Biol Rep 1993;17(3):167-183. ##Jiang T, Zhang J, Liang D. Structure and function of chromophores in R-phycoerythrin at 1.9 A resolution. Proteins 1999;34(2):224-231. ##Wang W, Singh S, Zeng DL, King K, Nema S. Antibody structure, instability, and formulation. J Pharm Sci 2006;96(1):1-26. ##Barbet J, Machy P, Leserman LD. Monoclonal antibody covalently coupled to liposomes: specific targeting to cells. J Supramol Struct Cell Biochem 2004;16(3):243-258. ##

An Investigation into the Antifungal Property of Fabaceae using Bioinformatics Tools 2 2 Chemodiversity in plants provides sources of great value which might be helpful for finding new leads in drug discovery programs. Fabaceae as the third largest family of flowering plants was chosen to investigate its possible antifungal activity. In order to increase the effectiveness of the result, molecular similarity methods and chemical data were used. Twelve plants were selected from Fabaceae and collected from the North and South of Iran. Percolation method with 80% ethanol was used for extraction of collected plants. Antifungal activities of these extracts were determined using broth microdilution method against Candida albicans (C. albicans) ATCC 10231, Aspergillus fumigatus (A. fumigatus) AF 293 and Asperigillus niger (A. niger) ATCC 16404. Extracts with promising activity were screened for toxicity with larvae of Artemia salina (brine shrimp). Dalbergia sissoo, Lathyrus pratensis, Oreophysa microphyalla, Astragalus stepporum, Ebenus stellata, Sophora alopecuroides, Ammodendron persicum and Taverniera cuneifolia showed activity against at least one of the microorganisms used in this study. According to the results of our experiment, the extracts of these plants can be used for further investigation in therapeutic research. 93 100 Zahra Arabi Drug Design and Bioinformatics Unit, Medical Biotechnology Department, Biotechnology Research Center, Pasteur Institute Faculty of Biological Science, Shahid Beheshti University Drug Design and Bioinformatics Unit, Medical Biotechnology Department, Biotechnology Research Center, Pasteur Institute Faculty of Biological Science, Shahid Beheshti University IranIran Soroush Sardari Drug Design and Bioinformatics Unit, Medical Biotechnology Department, Biotechnology Research Center, Pasteur Institute Drug Design and Bioinformatics Unit, Medical Biotechnology Department, Biotechnology Research Center, Pasteur Institute Iran Antifungal ResearchBioinformaticsFabaceaePlant extracts 35.pdf Aliero AA, Grierson DS, Afolayan AJ. Antifungal activity of Solanum pseudocapsicum. Res J Bot 2006;1:129-133. ##Capoor MR, Nair D, Deb M, Verma PK, Srivastava L, Aggarwal P. Emergence of non-albicans Candida species and antifungal resistance in a tertiary care hospital. Jpn J Infect Dis 2005;58:344-348. ##Olalde Rangel JA. The systemic theory of living systems and relevance to CAM. Evid Based Complement Alternat Med 2005;2(1):13-18. ##Wojciechowski MF, Lavin M, Sandesrson MJ. A phylogeny of Legumes (Leguminosae) based on analysis of the plastid MATK gene resolves many well-supported subclades within the family. Am J Bot 2004;91:1846-1862. ##Wink, M. Evolution of secondary metabolites from an ecological and molecular phylogenetic perspective. Phytochemistry 2003;64(1):3-19. ##Abad MJ, Ansuategui M, Bermejo P. Active antifungal substances from natural sources. ARKIVOC 2007;7:116-145 ##Lovkova MY, Buzuk GN, Sokolova SM, Kliment' eva NI. Chemical features of medicinal plants. Appl Biochem Microbiol 2001;37(3):229-237. ##Cowan MM. Plant products as antimicrobial agents. Clin Microbiol Rev 1999;12(4):564-582. ##Verpoorte R. Exploration of nature’s chemodiversity: the role of secondary metabolites as leads in drug development. Drug Discov Today 1998;3(5): 232-238. ##Leite SP, Vieira JRC, Medeiros PL, Leite RMP, Lima VLM, Xavier HS, et al. Antimicrobial activity of Indigofera suffruticosa. Evid Based Complement Alternat Med 2006;3(2):261-265. ##Palombo EA. Traditional medicinal plant extracts and natural products with activity against oral bacteria: potential application in prevention and treatment of oral diseases. Evid Based Complement Alternat Med 2009;10:1-15. ##Coley PD, Heller MV, Aizprua R, Arauz B, Flores N, Correa M, et al. Using ecological criteria to design plant collection strategies for drug discovery. Front Ecol Environ 2003;1(8):421-428. ##Ronsted N, Savolainen V, Molgaard P, Jager AK. Phylogenetic selection of Narcissus species for drug discovery. Biochem Syst Ecol 2008;36(5-6): 417-422. ##Gasteiger J, Engel Th. Chemoinformatics. Weinheim: Wiley-VCH; 2003. ##Johann S, Pizzolatti MG, Donnici C, Resende MA. Antifungal properties of plants used in Brazilian traditional medicine against clinically relevant fungal pathogens. Braz J Microbiol 2007;38(4): 632-637. ##Scorzoni L, Benaducci T, Fusco Almeida AM, Siqueira Silva DH, Bolzani VS, Soares Mendes Gianinni MJ. The use of standard methodology for determination of antifungal activity of natural products against medical yeasts Candida sp. and Cryptococcus sp. Braz J Microbiol 2007;38(3):391-397. ##Ernst EJ, Rogers PD. Antifungal Agents: Methods and Protocols. Totowa, NJ: Humana Press Inc; 2005. ##McLaughlin JL, Rogers LL, Anderson JE. The use of biological assays to evaluate botanicals. Drug Inform J 1998;32:513-524. ##Jansen R, Cai Z, Raubeson, L, Daniell, H, dePamphilis CW, Leebens-Mack J, et al. Analysis of 81 genes from 64 plastid genomes resolves relationships in angiosperms and identifies genome-scale evolutionary patterns. Proc Natl Acad Sci USA 2007;104(49):19369-19374. ##Daylight.com: Simplified Molecular Input Line Entry System (SMILE) [Internet]. Aliso Viejo, CA; c2008. Available from: http://www.daylight.com/ smiles/. ##Yadav H, Yadav M, Jain S, Bhardwaj A, Singh V, Parkash O, et al. Antimicrobial property of a herbal preparation containing Dalbergia sissoo and Datura tramonium with cow urine against pathogenic bacteria. Int J Immunopathol Pharmacol 2008;21(4): 1013-1020. ##Robeson DJ, Ingham JL, Harborne B. Identification of two chromone phytoalexins in the sweet pea, Lathyrus odoratus. Phytochemistry 1980;19(10): 2171-2173. ##Ryu YB, Westwood IM, Kang NS, Kim HY, Kim JH, Moon YH, et al. Kurarinol, tyrosinase inhibitor isolated from the root of Sophora flavescens. Phytomedicine 2008;15(8):612-618. ##Sheridan RP, Kearsley SK. Why do we need so many chemical similarity search methods? Drug Discov Today 2002;7(17):903-911. ##Jassbi AJ, Zamanizadehnajari S, Aberoomand AP, Tahara S. Antibacterial diterpenoids from Astra-galus brachystachys. Z Naturforsch 2002;57:1016-1021. ##Pistelli L, Bertoli A, Lepori E, Morelli I, Panizzi L. Antimicrobial and antifungal activity of crude extracts and isolated saponins from Astragalus verrucosus. Fitoterapia 2002;73(4):336-339. ##Uyar Z, Boke N, Turkay E, Koz O, Yasa I, Kirmizigul S. Flavonoid glycosides and methylinositol from Ebenus haussknechtii. Nat Prod Res 2006; 20(11):999-1007. ##Keymanesh K, Hamedi J, Moradi S, Mohammadipanah F, Sardari S. Antibacterial, antifungal and toxicity of rare Iranian plants. Int J Pharmacol 2009;5(1):81-85. ##Ranganathan S, Balajee SA. Anti-cryptococcus activity of combination of extracts of Cassia alata and Ocimum sanctum. Mycoses 2000;43(7-8):299-301. ##Adwan G, Mhanna M. Synergistic effects of plant extracts and antibiotics on Staphylococcus aureus strains isolated from clinical specimens. Middle East J Sci Res 2008;3(3):134-139. ##

Anti-Arthritic Activity of Premna serratifolia Linn., Wood against Adjuvant Induced Arthritis 2 2 Adjuvant induced arthritis is a chronic crippling, skeleton-muscular disorder having nearest approximation to human rheumatoid arthritis for which there is currently no medicine available effecting a permanent cure. Even modern drugs used for the amelioration of the symptoms, offer only temporary relief and also produce severe side effects. In the indigenous system of medicine, wood of Premna serratifolia Linn., is reported to be useful in the treatment of arthritis. It is a large shrub, distributed throughout Asia, used against a wide variety of diseases. However, no systematic study has been reported regarding its anti-arthritic activity. This work was aimed at the scientific validation of the ethno-pharmacological claim about its anti-arthritic property. In the present study, anti-arthritic activity of ethanol extract of Premna serratifolia Linn., wood is done by Freund's adjuvant induced arthritis model. Loss in body weight during arthritis condition was corrected on treatment with ethanol extract and standard drug, indomethacin. Biochemical parameters such as hemoglobin content, total WBC, RBC, erythrocyte and sedimentation rate were also estimated. The ethanol extract at the dose of 300 mg/kg body weight inhibited the rat paw edema by 68.32% which is comparable with standard drug indomethacin 74.87% inhibition of rat paw edema after 21 days. The results of the current investigation concluded, ethanol extract of Premna serratifolia Linn., wood possess a significant anti-arthritic activity against adjuvant induced arthritis and justifying its therapeutic role in arthritic condition. The observed anti-arthritic activity may be due to the presence of phytoconstituents such as irridiod glycosides, alkaloids, phenolic compounds and flavonoids. 101 106 Rekha Rajendran Department of Pharmacognosy and Phytochemistry, Mohamed Sathak A. J. College of Pharmacy Department of Pharmacognosy and Phytochemistry, Mohamed Sathak A. J. College of Pharmacy India Ekambaram Krishnakumar Department of Pharmaceutical Biotechnology, Balaji Institute of Pharmaceutical Sciences Department of Pharmaceutical Biotechnology, Balaji Institute of Pharmaceutical Sciences India ArthritisFreund’s adjuvantIndomethacin 36.pdf The Wealth of India, Dictionary of Indian raw materials and industrial products, vol. 8, New Delhi: Council of Scientific and Industrial Research; 1972,240. ##Kapoor LD. CRC Handbook of Ayurvedic Medicinal Plants, Herbal Reference Library. Boca Raton London, New York Washington: CRC Press; 2001, 70. ##Rajendran R, Suseela L, Meenakshi Sundaram R, Saleem Basha N. 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Antimicrobial activity of crude extracts and fractions of Premna serratifolia Linn., root. Medicinal Plants-Int J Phytomed Rel Indus 2010;2(1):[1p.]. ##Hazeena BV, Sadique J. Long term effect of herbal drug Withania somnifera on adjuvant induced arthritis in rats. Indian J Exp Biol 1988;26(11):877-882. ##Henry AN, Kumari GR, Chitra V. Flora of Tamil Nadu. Botanical survey of India southern circle (Vol 2). Coimbatore;1987,167. ##Harbone JB. Phytochemical methods, a guide to modern techniques of plant analysis. 2nd ed. London, New York: Chapman and Hal;1973,90. ##Ecobichon DJ. The basis of toxicology testing. 2nd ed. New York: CRC press; 1999,43-46. ##Newbould BB. Chemotherapy of arthritis induced in rats by mycobacterial adjuvant. Br J Pharmacol Chemother 1963;21(1):127-136. ##Carl MP. Experimental joint disease observations on adjuvant induced arthritis. J Chronic Dis 1963; 16(8):863-864. ##Singh S, Majumdar DK. Effect of fixed oils of Ocimum sanctum against experimentally induced arthritis and joint edema in laboratory animals. J Pharmacogn 1996;34(3):218-222. ##Eric GB, Lawrence JL. Rheumatoid arthritis and its therapy, The Textbook of therapeutic drugs and disease management. 6th ed. Baltimore: Williams and Wilkins Company; 1996,579-595. ##William JK. Arthritis and allied condition, A Textbook of rheumatology (Vol. 1). 3rd ed. Baltimore: Tokyo: A Waverly Company; 1996,207-226. ##Winder CV, Lembke LA, Stephens MD. Comparative bioassay of drugs in adjuvant induced arthritis in rats: flufenamic acid, mefenamic acid and phenylbutazone. Arthritis Rheum 2005;12(5):472-482. ##Walz DT, Dimartino MJ, Misher A. Adjuvant induced arthritis in rats.II. Drug effects on physiologic, biochemical and immunologic parameters. J Pharmacol Exp Ther 1971;178(1):223-231. ##Somasundaran S, Sadique J, Subramoniam A. Influence of extra-intestinal inflammation on the in vitro absorption of 14C-glucose and the effects of anti-inflammatory drugs in the jejunum of rats. Clin Exp Pharmacol Physiol 1983;10(2):147-152. ##Somasundaran S, Sadique J, Subramoniam A. In vitro absorption of [14C] leucine during inflammation and the effect of anti-inflammatory drugs in the jejunum of rats. Biochem Med 1983;29(2):259-264. ##Elmali N, Esenkaya I, Harma A, Ertem K, Turkoz Y, Mizrak B. Effect of resveratrol in experimental osteoarthritis in rabbits. Inflamm Res 2005;54(4): 158-162. ##Choi EJ, Bae SC, Yu R, Youn J, Sung MK. Dietary vitamin E and quercetin modulate inflammatory responses of collagen-induced arthritis in mice. J Med Food 2009;12(4):770-775. ##Amresh G, Singh PN, Rao ChV. Antinociceptive and antiarthritic activity of Cissampelos pareira roots. J Ethnopharmacol 2007;111(3):531-536. ##

The Effect of Biopsy During Precompacted Morula Stage on Post Vitrification Development of Blastocyst Derived Bovine Embryos 2 2 Improvements on embryo micromanipulation techniques led to the use of embryo biopsy in commercial embryo transfer programs for genetic analysis of preimplantation bovine embryos. The aim of this study was to evaluate the quality of bovine blastocyst derived from embryos biopsied at different precompacted morulae stages by assessment of cryosurvivability of the resulting blastocysts. The in vitro produced bovine embryos were subjected to biopsy at days 2, 3, and 4 post-insemination with different cell numbers (4 to 16-cells). Embryo cell biopsy was carried out in a 100 ?l drop of H-SOF following pronase drilling by aspiration of one blastomere. The biopsied embryos were then cultured in SOFaaBSA co-cultured with oviduct cells-monolayer until blastocyst formation. The blastocysts were cryopreserved at room tempera-ture after exposure of equilibration (glycerol 1.4 M for 5 min and then glycerol 1.4 M and ethylene glycol 3.6 M for 5 min) and vitrification solutions (3.4 M glycerol and 4.6 M ethylene glycol). The blastocysts were loaded into the center of 0.25 ml straws separated by air bubbles from 2 columns of sucrose 0.5 M and plunged immediately into liquid nitrogen. There was no significant difference in cryosurvivability of vitrified-warmed blastocysts derived form biopsied embryos at different pre-compacted morula stages. The quality of biopsy derived blastocysts was identical to that of non-biopsy derived ones in terms of post vitrifcation survival and hatching rates. In conclusion there was no preference between different times of embryo biopsy at precompacted morula stages in term of cryosurvivability of biopsy derived bovine blastocysts. 107 111 Abolfazl Shirazi Research Institute of Animal Embryo Technology, Shahrekord UniversityReproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Research Institute of Animal Embryo Technology, Shahrekord UniversityReproductive Biotechnology Research Center, Avicenna Research Institute, ACECR IranIran Sara Borjian Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Ebrahim Ahmadi Research Institute of Animal Embryo Technology, Shahrekord University Research Institute of Animal Embryo Technology, Shahrekord University Iran Hassan Nazari Research Institute of Animal Embryo Technology, Shahrekord University Research Institute of Animal Embryo Technology, Shahrekord University Iran Banafsheh Heidari Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran BovineCryopreservationEmbryo ResearchFertilization in Vitro 37.pdf Bowen RA, Reed ML, Schnieke A, Seidel GE Jr, Stacey A, Thomas WK, Kajikawa O. Transgenic cattle resulting from biopsied embryos: Expression of c-ski in a transgenic calf. Biol Reprod 1994; 50(3):664-668. ##Hyttinen JM, Peura T, Tolvanen M, Aalto J, Jänne J. Detection of microinjected genes in bovine preimplantation embryos with combined DNA diges-tion and polymerase chain reaction. Mol Reprod Dev 1996;43(2):150-157. ##Agrawala PL, Wagner VA, Geldermann H. Sex determination and milk protein genotyping of preimplantation stage bovine embryos using multiplex PCR. Theriogenology 1992;38:969-978. ##Handyside AH, Kontogianni EH, Hardy K, Winston RML. Pregnancy from human biopsied preimplantation embryos sexed by Y-specific DNA amplification. Nature 1990;344:768-770. ##Herr CM, Reed KC. Micromanipulation of bovine embryos for sex determination. Theriogenolgy 1991;35:45-54. ##Gustafsson H, Jaakma Ü, Shamsuddin M. Viability of fresh and frozen-thawed biopsied bovine embryos. Acta Vet Scand 1994;35(3):217-222. ##Thibier M, Nibart M. The sexing of bovine embryos in the field. Theriogenology 1995;43(1):71-80. ##Urszula B, Lutjen KJ, O'Neill C. Assessment of the viability and pregnancy potential of mouse embryos biopsied at different preimplantation stages of development. Hum Reprod 1990;5(2):203-208. ##Naitana S, Loi P, Ledda S, Cappai P, Dattena M, Bogliolo L, et al. Effect of biopsy and vitrification on in vitro survival of ovine embryos at different stages of development. Theriogenology 1996;46(5): 813-824. ##Chesné P, Heyman Y, Chupin D, Procureur R, Ménézo Y. Freezing cattle demi-embryos: influence of a period of culture between splitting and freezing on survival. Theriogenology 1987;27:218. ##Shirazi A, Borjian S, Nazari H, Ahmadi E, Heidari B, Bahiraee A. ¬¬Effects of timing on cell biopsy from pre-compacted morula stage bovine embryos on subsequent embryonic development. J Reprod Infertil 2010;11(1):25-32. ##Shirazi A, Soleimani M, Karimi M, Nazari H, Ahmadi E, Heidari B. Vitrification of in vitro produced ovine embryos at various developmental stages using two methods. Cryobiology 2010;60(2):204-210. ##Ali J, Shelton JN. Successful vitrification of day-6 sheep embryos. J Reprod Fertil 1993;99:65-70. ##Szell AZ, Windsor DP. Survival of vitrified sheep embryos in vitro and in vivo. Theriogenology 1994;42:881-889. ##Suzuki T, Saha S, Sumantri C, Takagi M, Boediono A. The influence of polyvinyl pyrolidone on freezing of bovine IVF blastocysts following biopsy. Cryobiology 1995;32(6):505-510. ##Vajta G, Holm P, Greve T, Callesen H. Cumulative efficiency of biopsy, vitrification and in straw dilution in a bovine in vitro embryo production system. Theriogenology 1996;45(1):162. ##Agca Y, Monson, RL, Northey DL, Peschel DE, Schaefer DM, Rutledge JJ. Normal calves from transfer of biopsied, sexed and vitrified IVP bovine embryos. Theriogenology 1998;50(1):129-145. ##Lopes RFF, Forell F, Oliveira ATD, Rodrigues JL. Spliting and biopsy for bovine embryo sexing under fields conditions. Theriogenology 2001;56 (9):1383-1392. ##Carroll J, Depypere H, Matthews CD. Freeze–thaw-induced changes of the zona pellucida explains decreased rates of fertilization in frozen–thawed mouse oocytes. J Reprod Fertil 1990;90: 547-553. ##Ghetler Y, Skutelsky B, Ben Nun I, Ben Dor L, Amihai D, Shalgi R. Human oocyte cryopreservation and the fate of cortical granules. Fert Steril 2006;86(1):210-216. ##