Future directions for translation of tissue engineering products into clinic 2 2 At present, Iran has been known one of the up-warding countries in the world in regenerative medicine using stem cells therapy. In fact, the outcomes of some clinical trials on stem cell therapy of myocardial infarction, vitiligo, decompensated cirrhosis, and osteoarthritis narrate the feasibility of stem cell-based therapy for treatment of human diseases 1,2. However, in a similar manner with global configuration, the commercialization and translation of tissue engineering products into clinical phase has been restricted. It might be due to weak collaboration of different specialties for technology transfer of the multidisciplinary projects of tissue engineering field into clinical phase. Basic tissue engineers mostly prefer elegant studies, whereas physicians have tendency to solve medical problems with products indicating efficiency, easy to use, and cost benefit. Actually, a surgeon encountered with a dilemma between a partially effective tissue-engineered product that is both expensive and difficult to apply and a more traditional approach may choose the latter option. Therefore, a coherent teamwork between basic sciences and medicine as well as acquisition of competent knowledge about target tissue is necessary to conduct tissue engineering in the clinic. Moreover, it should be considered that in developing countries including Iran the high cost of high-tech biomedical research necessitates government investment 3. Currently, the policy makers have established some action plans to support of science-based companies financially. This is a suitable opportunity to ligature basic research and market for commercialization of tissue engineering products. However, because private investors beyond academic laboratories should provide financing of tissue engineering products, incentive of private companies for investment should not be neglected. It is notable that tissue-engineered products will fulfill small market size unless they could indicate much superior results than competitive alternatives. It is noticeable that tissue engineers should determine the requirements of community and develop strategies to penetrate the products into clinic. Indeed, the communication between scientists and policy makers should be increased to better definition of national research priorities. On the other hand, considering local necessities and natural resources should be rather than subjective experts’ notions or international superiorities. Finally, ethical and legal regulations should be actually defined that indubitably make great profits to the society. 103 103 Somaieh Kazemnejad Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Editorial 240.pdf Mobini S, Khanmohammadi M, Heidari-Vala H, Samadikuchaksaraei A, Moshiri A, Kazemnejad S. Tissue Engineering and Regenerative Medicine in Iran: Current State of Research and Future Outlook. Mol Biotechnol 2015;57(7):589-605.##Gheisari Y, Baharvand H, Nayernia K, Vasei M. Stem cell and tissue engineering research in the Islamic Republic of Iran. Stem Cell Rev 2012;8(3):629-639.##Samadikuchaksaraei A, Mousavizadeh K. High-tech biomedical research: lessons from Iran's experience. Biomed Eng Online 2008;7:17.##

Development of a Single Stranded DNA Aptamer as a Molecular Probe for LNCap Cells Using Cell-SELEX 2 2 Background: Nowadays, highly specific aptamers generated by cell SELEX technology (systematic evolution of ligands by exponential enrichment) are being applied for early detection of cancer cells. Prostate Specific Membrane Antigen (PSMA), over expressed in prostate cancer, is a highly specific marker and therefore can be used for diagnosis of the prostate cancer cells. The aim of the present study was to select single-stranded DNA aptamers against LNCap cells highly expressing PSMA, using cell–SELEX method which can be used as a diagnostic tool for the detection of prostate cancer cells. Methods: After 10 rounds of cell-SELEX, DNA aptamers were isolated against PSMA using LNCaP cells as a target and PC-3 cell lines for counter SELEX. Five DNA aptamers with more than 70% affinity were selected up on flow cytometry analysis of positive clones. Results: Dissociation constants of two selected sequences (A12-B1) were estimated in the range of 33.78±3.77 and 57.49±2.214 pmol, respectively. Conserved secondary structures of A12 and B1 sequences suggest the necessity of these structures for binding with high affinity to native PSMA. Comparison of the secondary structures of our isolated aptamers and aptamer A10 obtained by protein SELEX showed similar stem-loop structures which could be responsible for the recognition of PSMA on LNCap cell surface. Conclusion: Our results indicated that selected aptamers may turn out to be ideal candidates for the development of a detection tool and also can be used in targeted drug delivery for future smart drugs. 104 111 Faezeh Almasi Department of Biotechnology, College of Science, Tehran University Department of Biotechnology, College of Science, Tehran University Iran Seyed Latif Mousavi Gargari Department of Biology, Shahed University Department of Biology, Shahed University Iran Fatemeh Bitaraf Department of Biology, Shahed University Department of Biology, Shahed University Iran Samaneh Rasoulinejad Department of Biology, Shahed University Department of Biology, Shahed University Iran Cell-SELEXDNA aptamerExonucleasesProstate specific membrane antigen 242.pdf Siegel R, Naishadham D, Jemal A. Cancer statistics, 2013. CA Cancer J Clin 2013;63(1):11-30.##Lepor A, Catalona WJ, Loeb S. The prostate health index: its utility in prostate cancer detection. Urol Clin North Am 2016;43(1):1-6.##Bacich DJ, Pinto JT, Tong WP, Heston WD. Cloning, expression, genomic localization and enzymatic activities of the mouse homolog of prostate-specific membrane antigen/NAALADase/folate hydrolase. Mamm Genome 2001;12(2):117-123.##Beckett ML, Cazares LH, Vlahou A, Schellhammer PF, Wright GL Jr. Prostate-specific membrane antigen levels in sera from healthy men and patients with benign prostate hyperplasia or prostate cancer. Clin Cancer Res 1999;5(12):4034-4040.##Rajasekaran AK, Anilkumar G, Christiansen JJ. Is prostate-specific membrane antigen a multifunctional protein? Am J Physiol Cell Physiol 2005;288(5):C975-981.##Cibiel A, Pestourie C, Ducongé F. In vivo uses of aptamers selected against cell surface biomarkers for therapy and molecular imaging. Biochimie 2012;94(7):1595-1606.##Nery AA, Wrenger C, Ulrich H. Recognition of biomarkers and cell-specific molecular signatures: aptamers as capture agents. J Sep Sci 2009;32(10):1523-1530.##Bitaraf FS, Rasooli I, Mousavi Gargari SL. DNA aptamers for the detection of Haemophilus influenzae type b by cell SELEX. Eur J Clin Microbiol Infect Dis 2016;35(3):503-510.##Ohuchi S. Cell-SELEX Technology. Biores Open Access 2012;1(6):265-272.##Lupold SE, Hicke BJ, Lin Y, Coffey DS. Identification and characterization of nuclease-stabilized RNA molecules that bind human prostate cancer cells via the prostate-specific membrane antigen. Cancer Res 2002;62(14):4029-4033.##Dassie JP, Liu XY, Thomas GS, Whitaker RM, Thiel KW, Stockdale KR, et al. Systemic administration of optimized aptamer-siRNA chimeras promotes regression of PSMA-expressing tumors. Nat Biotechnol 2009;27(9):839-849.##Ni X, Zhang Y, Ribas J, Chowdhury WH, Castanares M, Zhang Z, et al. Prostate-targeted radiosensitization via aptamer-shRNA chimeras in human tumor xenografts. J Clin Invest 2011;121(6):2383-2390.##Chu TC, Marks JW 3rd, Lavery LA, Faulkner S, Rosenblum MG, Ellington AD, et al. Aptamer: toxin conjugates that specifically target prostate tumor cells. Cancer Res 2006;66(12):5989-5992.##Dhar S, Gu FX, Langer R, Farokhzad OC, Lippard SJ. Targeted delivery of cisplatin to prostate cancer cells by aptamer functionalized Pt(IV) prodrug-PLGA-PEG nanoparticles. Proc Natl Acad Sci USA 2008;105(45):17356-17361.##Kasten BB, Liu T, Nedrow-Byers JR, Benny PD, Berkman CE. Targeting prostate cancer cells with PSMA inhibitor-guided gold nanoparticles. Bioorg Med Chem Lett 2013;23(2):565-568.##Xu W, Siddiqui IA, Nihal M, Pilla S, Rosenthal K, Mukhtar H, et al. Aptamer-conjugated and doxorubicin-loaded unimolecular micelles for targeted therapy of prostate cancer. Biomaterials 2013;34(21):5244-5253.##Wang AZ, Bagalkot V, Vasilliou CC, Gu F, Alexis F, Zhang L, et al. Superparamagnetic iron oxide nanoparticle-aptamer bioconjugates for combined prostate cancer imaging and therapy. ChemMedChem 2008;3(9):1311-1315.##Ben Jemaa A, Bouraoui Y, Sallami S, Banasr A, Ben Rais N, Ouertani L, et al. Co-expression and impact of prostate specific membrane antigen and prostate specific antigen in prostatic pathologies. J Exp Clin Cancer Res 2010;29:171.##Tewari A, ed. Prostate cancer: A comprehensive perspective. 1st ed. London: Springer-Verlag; 2013. 1110 p.##Actis P, Rogers A, Nivala J, Vilozny B, Seger RA, Jejelowo O, et al. Reversible thrombin detection by aptamer functionalized STING sensors. Biosens Bioelectron 2011;26(11):4503-4507.##Meyer C, Hahn U, Rentmeister A. Cell-specific aptamers as emerging therapeutics. J Nucleic Acids 2011;2011:904750.##Zare H, Rajabibazl M, Rasooli I, Ebrahimizadeh W, Bakherad H, Ardakani LS, et al. Production of nanobodies against prostate-specific membrane antigen (PSMA) recognizing LnCaP cells. 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The Cytotoxicity of Dacarbazine Potentiated by Sea Cucumber Saponin in Resistant B16F10 Melanoma Cells through Apoptosis Induction 2 2 Background: Malignant melanoma is a highly aggressive malignant melanocytic neoplasm which resists against the most conventional therapies. Sea cucumber as one of marine organisms contains bioactive compounds such as polysaccharide, terpenoid and other metabolites which have anti-cancer, anti-tumor, anti-inflammatory and antioxidant properties. The present study was designed to investigate the anticancer potential of saponin extracted from sea cucumber Holothuria leucospilata alone and in combination with dacarbazine on B16F10 melanoma cell line. Methods: The B16F10 cell line was treated with different concentrations of saponin (0, 4, 8, 12, 16, 20 µg/ml), dacarbazine (0, 1200, 1400, 1600, 1800, 2000 µg/ml) and co-administration of saponin-dacarbazine (1200 da+8 sp, 1200 da+4 sp) for 24 and 48 hr and the cytotoxic effect was examined by MTT, DAPI, acridine orange/propodium iodide, flow cytometry and caspase colorimetric assay. Results: The results exhibited that sea cucumber saponin, dacarbazine, and co-administration of saponin-dacarbazine inhibited the proliferation of melanoma cells in a dose and time dependent manner with IC50 values of 10, 1400 and 4+1200 µg/ml, respectively. Morphological observation of DAPI and acridine orange/propodium iodide staining documented typical characteristics of apoptotic cell death. Flow cytometry assay indicated accumulation of IC50 treated cells in sub-G1 peak. Additionally, saponin extracted induced intrinsic apoptosis via up-regulation of caspase-3 and caspase-9. Conclusion: These results revealed that the saponin extracted from sea cucumber as a natural anti-cancer compound may be a new treatment modality for metastatic melanoma and the application of sea cucumber saponin in combination with dacarbazine demonstrated the strongest anti-cancer activity as compared with the drug alone. 112 119 Javad Baharara Department of Biology, Research Center for Applied Biology, Mashhad Branch, Islamic Azad University Department of Biology, Research Center for Applied Biology, Mashhad Branch, Islamic Azad University Iran Elaheh Amini Department of Animal Biology, Faculty of Biological Sciences, Kharazmi University Department of Animal Biology, Faculty of Biological Sciences, Kharazmi University Iran Najme Nikdel Research Center for Applied Biology, Mashhad Branch, Islamic Azad University Research Center for Applied Biology, Mashhad Branch, Islamic Azad University Iran Farzaneh Salek-Abdollahi Research Center for Applied Biology, Mashhad Branch, Islamic Azad University Research Center for Applied Biology, Mashhad Branch, Islamic Azad University Iran ApoptosisDacarbazineMelanomaSaponinsSea cucumbers 243.pdf Khoobchandani M, Ganesh N, Gabbanini S, Valgimigli L, Srivastava MM. Phytochemical potential of Eruca sativa for inhibition of melanoma tumor growth. Fitoterapia 2011;82(4):647-653.##Salma Y, Lafont E, Therville N, Carpentier S, Bonnafé MJ, Levade T, et al. The natural marine anhydrophytosphingosine, Jaspine B, induces apoptosis in melanoma cells by interfering with ceramide metabolism. Biochem Pharmacol 2009;78(5):477-485.##D'Orazio J, Jarrett S, Amaro-Ortiz A, Scott T. UV radiation and the skin. Int J Mol Sci 2013;14(6):12222-12248.##Chinembiri TN, du Plessis LH, Gerber M, Hamman JH, du Plessis J. Review of natural compounds for potential skin cancer treatment. Molecules 2014;19(8):11679-11721.##Zigler M, Villares GJ, Lev DC, Melnikova VO, Bar-Eli M. Tumor immunotherapy in melanoma: strategies for overcoming mechanisms of resistance and escape. Am J Clin Dermatol 2008;9(5):307-311.##Grossman D, Altieri DC. Drug resistance in melanoma: mechanisms, apoptosis, and new potential therapeutic targets. 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Triterpenoids of marine origin as anti-cancer agents. Molecules 2013;18(7):7886-7909.##Bahrami Y, Zhang W, Franco C. Discovery of novel saponins from the viscera of the sea cucumber Holothuria lessoni. Mar Drugs 2014;12(5):2633-2667.##Esmat AY, Said MM, Soliman AA, El-Masry KS, Badiea EA. Bioactive compounds, antioxidant potential, and hepatoprotective activity of sea cucumber (Holothuria atra) against thioacetamide intoxication in rats. Nutrition 2013;29(1):258-267.##Wijesinghe WA, Jeon YJ, Ramasamy P, Wahid ME, Vairappan CS. Anticancer activity and mediation of apoptosis in human HL-60 leukaemia cells by edible sea cucumber (Holothuria edulis) extract. Food Chem 2013;139(1-4):326-331.##Janakiram NB, Mohammed A, Rao CV. Sea cucumbers metabolites as potent anti-cancer agents. Mar Drugs 2015;13(5):2909-2923.##He NW, Zhao Y, Guo L, Shang J, Yang XB. Antioxidant, antiproliferative, and pro-apoptotic activities of a saponin extract derived from the roots of Panax notoginseng (Burk.) F.H. Chen. J Med Food 2012;15(4):350-359.##Soltani M, Parivar K, Baharara J, Kerachian MA, Asili J. Putative mechanism for apoptosis-inducing properties of crude saponin isolated from sea cucumber (Holothuria leucospilota) as an antioxidant compound. Iran J Basic Med Sci 2015;18(2):180-187.##Hu XQ, Wang YM, Wang JF, Xue Y, Li ZJ, Nagao K, et al. Dietary saponins of sea cucumber alleviate orotic acid-induced fatty liver in rats via PPARalpha and SREBP-1c signaling. Lipids Health Dis 2010;9:25.##Lewandowska U, Gorlach S, Owczarek K, Hrabec E, Szewczyk K. Synergistic interactions between anticancer chemotherapeutics and phenolic compounds and anticancer synergy between polyphenols. Postepy Hig Med Dosw (Online) 2014;68:528-540.##Swift LH, Golsteyn RM. Genotoxic anti-cancer agents and their relationship to DNA damage, mitosis, and checkpoint adaptation in proliferating cancer cells. Int J Mol Sci 2014;15(3):3403-3431.##Bojo ZP, Deaño CD, Jacinto SD, Concepcion GP. Synergistic in vitro cytotoxicity of adociaquinone B and heptyl prodigiosin against MCF-7 breast cancer cell line. Philipp Sci Lett 2010;3(2):48-58.##Hammerová J, Uldrijan S, Táborská E, Slaninová I. Benzo[c]phenanthridine alkaloids exhibit strong anti-proliferative activity in malignant melanoma cells regardless of their p53 status. J Dermatol Sci 2011;62(1):22-35.##Oktem G, Uysal A, Oral O, Sezer ED, Olukman M, Erol A, et al. Resveratrol attenuates doxorubicin-induced cellular damage by modulating nitric oxide and apoptosis. Exp Toxicol Pathol 2012;64(5):471-479.##Tabolacci C, Lentini A, Mattioli P, Provenzano B, Oliverio S, Carlomosti F, et al. Antitumor properties of aloe-emodin and induction of transglutaminase 2 activity in B16-F10 melanoma cells. Life Sci 2010;87(9-10):316-324.##Zhang F, Zhang Z, Liu Z. Effects of Huaier aqueous extract on proliferation and apoptosis in the melanoma cell line A875. Acta Histochem 2013;115(7):705-711.##Fujiki H, Suganuma M. 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Neuroprotective Effects of Herbal Extract (Rosa canina, Tanacetum vulgare and Urtica dioica) on Rat Model of Sporadic Alzheimer’s Disease 2 2 Background: Sporadic Alzheimer’s Disease (SAD) is caused by genetic risk factors, aging and oxidative stresses. The herbal extract of Rosa canina (R. canina), Tanacetum vulgare (T. vulgare) and Urtica dioica (U. dioica) has a beneficial role in aging, as an anti-inflammatory and anti-oxidative agent. In this study, the neuroprotective effects of this herbal extract in the rat model of SAD was investigated. Methods: The rats were divided into control, sham, model, herbal extract -treated and ethanol-treated groups. Drug interventions were started on the 21st day after modeling and each treatment group was given the drugs by intraperitoneal (I.P.) route for 21 days. The expression levels of the five important genes for pathogenesis of SAD including Syp, Psen1, Mapk3, Map2 and Tnf-α were measured by qPCR between the hippocampi of SAD model which were treated by this herbal extract and control groups. The Morris Water Maze was adapted to test spatial learning and memory ability of the rats. Results: Treatment of the rat model of SAD with herbal extract induced a significant change in expression of Syp (p=0.001) and Psen1 (p=0.029). In Morris Water Maze, significant changes in spatial learning seen in the rat model group were improved in herbal-treated group. Conclusion: This herbal extract could have anti-dementia properties and improve spatial learning and memory in SAD rat model. 120 125 Parvaneh Daneshmand Genetic Research Center, University of Social Welfare and Rehabilitation Sciences Genetic Research Center, University of Social Welfare and Rehabilitation Sciences Iran Kioomars Saliminejad Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Marzieh Dehghan Shasaltaneh Laboratory of Neuro-organic Chemistry, Institute of Biochemistry and Biophysics (IBB), University of Tehran Laboratory of Neuro-organic Chemistry, Institute of Biochemistry and Biophysics (IBB), University of Tehran Iran Koorosh Kamali Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Gholam Hossein Riazi Laboratory of Neuro-organic Chemistry, Institute of Biochemistry and Biophysics (IBB), University of Tehran Laboratory of Neuro-organic Chemistry, Institute of Biochemistry and Biophysics (IBB), University of Tehran Iran Reza Nazari Laboratory of Neuro-organic Chemistry, Institute of Biochemistry and Biophysics (IBB), University of Tehran Laboratory of Neuro-organic Chemistry, Institute of Biochemistry and Biophysics (IBB), University of Tehran Iran Pedram Azimzadeh Basic and Molecular Epidemiology of Gastrointestinal Disorders Research Center, Shahid Beheshti University of Medical Sciences Basic and Molecular Epidemiology of Gastrointestinal Disorders Research Center, Shahid Beheshti University of Medical Sciences Iran Hamid Reza Khorram Khorshid Genetic Research Center, University of Social Welfare and Rehabilitation Sciences Genetic Research Center, University of Social Welfare and Rehabilitation Sciences Iran Alzheimer diseaseGene expressionHerbal extract 244.pdf Diwu YC, Tian JZ, Shi J. 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Inhibitory Effects of Some Carbohydrates on Nano-Globular Aggregation of both Normal and Glycated Albumin 2 2 Background: Protein aggregation is one of the important, common and troubling problems in biotechnology, pharmaceutical industries and amyloid-related disorders. Methods: In the present study, the inhibitory effects of some carbohydrates (alginate, β-cyclodextrin and trehalose) on the formation of nano-globular aggregates from normal (HSA) and glycated (GHSA) human serum albumin were studied; when the formation of aggregates was induced by the simultaneous heating and addition of dithiotheritol. For the investigations, the biophysical methods of UV-vis spectrophotometry, circular dichroism spectroscopy, transmission electron microscopy and tensiometry were employed. Results: The effect of inhibitory mechanism of these inhibitors on the aggregation of HSA and GHSA was expressed and compared together. Conclusion: The results showed that the nucleus formation step of the aggregation process of HSA and GHSA was different in the presence of alginate (compared to β-cyclodextrin and trehalose). The inhibition efficiencies of the carbohydrates on the aggregate formation of HSA and GHSA were different, arising from the differences in the hydrophobicities of HSA and GHSA, and also, the differences between HSA- and GHSA-carbohydrate interactions. 126 132 Ali Akbar Moosavi-Movahedi Institute of Biochemistry and Biophysics, University of Tehran Institute of Biochemistry and Biophysics, University of Tehran Iran Naghmeh Sattarahmady Department of Medical Physics, Faculty of Medicine, Shiraz University of Medical SciencesNanobiology and Nanomedicine Research Center, Shiraz University of Medical SciencesPharmaceutical Sciences Research Center, Shiraz University of Medical Sciences Department of Medical Physics, Faculty of Medicine, Shiraz University of Medical SciencesNanobiology and Nanomedicine Research Center, Shiraz University of Medical SciencesPharmaceutical Sciences Research Center, Shiraz University of Medical Sciences IranIranIran Esmaeil Sharifi Institute of Biochemistry and Biophysics, University of Tehran Institute of Biochemistry and Biophysics, University of Tehran Iran Hossein Heli Nanobiology and Nanomedicine Research Center, Shiraz University of Medical Sciences Nanobiology and Nanomedicine Research Center, Shiraz University of Medical Sciences Iran AlbuminAlginateProtein aggregationTrehalose 245.pdf Bagger HL, Øgendal LH, Westh P. 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Fold Des 1998;3(1):R9-23.##Mierzeiewska D, Mitrowska P, Rudnicka B, Kubicka E, Kostyra H. Effect of non-enzymatic glycosylation of pea albumins on their immunoreactive properties. Food Chem 2008;111(1):127-131.##Lee MJ, Fennema OR. Ability of cyclodextrins to inhibit aggregation of beta-casein. J Agric Food Chem 1991;39(1):17-21.##Richards AB, Krakowka S, Dexter LB, Schmid H, Wolterbeek AP, Waalkens-Berendsen DH, et al. Trehalose: a review of properties, history of use and human tolerance, and results of multiple safety studies. Food Chem Toxicol 2002;40(7):871-898.##Wyatt GR, Kale GF. The chemistry of insect hemolymph. II. Trehalose and other carbohydrates. J Gen Physiol 1957;40(6):833-847.##Stephen AM, Phillips GO, Williams PA. Food Polysaccharides and Their Applications. New York, USA: CRC Press; 2006. 625p.##Hurst R, Bao Y, Ridley S, Williamson G. Phospholipid hydroperoxide cysteine peroxidase activity of human serum albumin. 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Effect of Removal of Spermatogonial Stem Cells (SSCs) from In Vitro Culture on Gene Expression of Niche Factors in Bovine 2 2 Background: Niche cells, regulating Spermatogonial Stem Cells (SSCs) fate are believed to have a reciprocal communication with SSCs. The present study was conducted to evaluate the effect of SSC elimination on the gene expression of Glial cell line-Derived Neurotrophic Factor (GDNF), Fibroblast Growth Factor 2 (FGF2) and Kit Ligand (KITLG), which are the main growth factors regulating SSCs development and secreted by niche cells, primarily Sertoli cells. Methods: Following isolation, bovine testicular cells were cultured for 12 days on extracellular matrix-coated plates. In the germ cell-removed group, the SSCs were removed from the in vitro culture using differential plating; however, in the control group, no intervention in the culture was performed. Colony formation of SSCs was evaluated using an inverted microscope. The gene expression of growth factors and spermatogonia markers were assessed using quantitative real time PCR. Results: SSCs colonies were developed in the control group but they were rarely observed in the germ cell-removed group; moreover, the expression of spermatogonia markers was detected in the control group while it was not observed in the germ cell-removed group, substantiating the success of SSCs removal. The expression of Gdnf and Fgf2 was greater in the germ cell-removed than control group (p<0.05), whereas the expression of Kitlg was lower in the germ cell-removed than control group (p<0.05). Conclusion: In conclusion, the results revealed that niche cells respond to SSCs removal by upregulation of GDNF and FGF2, and downregulation of KITLG in order to stimulate self-renewal and arrest differentiation. 133 138 Vahid Akbarinejad Department of Theriogenology, Faculty of Veterinary Medicine, University of TehranTheriogenology Association, Faculty of Veterinary Medicine, University of Tehran Department of Theriogenology, Faculty of Veterinary Medicine, University of TehranTheriogenology Association, Faculty of Veterinary Medicine, University of Tehran IranIran Parviz Tajik Theriogenology Association, Faculty of Veterinary Medicine, University of Tehran Theriogenology Association, Faculty of Veterinary Medicine, University of Tehran Iran Mansooreh Movahedian Department of Anatomical Sciences, Faculty of Medical Sciences, Tarbiat Modares University Department of Anatomical Sciences, Faculty of Medical Sciences, Tarbiat Modares University Iran Reza Youssefi Department of Theriogenology, Faculty of Veterinary Medicine, University of TehranTheriogenology Association, Faculty of Veterinary Medicine, University of Tehran Department of Theriogenology, Faculty of Veterinary Medicine, University of TehranTheriogenology Association, Faculty of Veterinary Medicine, University of Tehran IranIran BovineGene expressionStem cells 246.pdf Oatley JM, Brinster RL. 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Spermatogonial stem cell biology in the bull: development of isolation, culture, and transplantation methodologies and their potential impacts on cattle production. Soc Reprod Fertil Suppl 2010;67:133-143.##Shafiei Sh, Tajik P, Ghasemzadeh-nava H, Movahedin M, Talebkhan Garoussi M, Qasemi-Panahi B, et al. Isolation of bovine spermatogonial cells and co-culture with prepubertal sertoli cells in the presence of colony stimulating factor-1. Iran J Vet Med 2013;7(2):83-90.##He Z, Kokkinaki M, Jiang J, Dobrinski I, Dym M. Isolation, characterization, and culture of human spermatogonia. Biol Reprod 2010;82(2):363-372.##Reding SC, Stepnoski AL, Cloninger EW, Oatley JM. THY1 is a conserved marker of undifferentiated spermatogonia in the pre-pubertal bull testis. Reproduction 2010;139(5):893-903.##Izadyar F, Den Ouden K, Creemers LB, Posthuma G, Parvinen M, De Rooij DG. Proliferation and differentiation of bovine type A spermatogonia during long-term culture. 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The Effect of Media Supplementation with Angiotensin on Developmental Competence of Ovine Embryos Derived from Vitrified-warmed Oocytes 2 2 Background: This study was aimed to assess the effects of angiotensin II (Ang II) supplementation to the In Vitro Maturation (IVM) and In Vitro Culture (IVC) media of vitrified-warmed ovine oocytes on their developmental competence and expression of Na+/K+/ATPase in resulting embryos. Methods: The slaughterhouse-derived immature oocytes (n=1069) were randomly distributed into four experimental groups: groups I and II) IVM/IVF and IVC of fresh and vitrified oocytes without angiotensin supplementation (Control-Fresh and Control-Vit groups, respectively); group III) IVM of vitrified oocytes in the presence of Ang II followed by IVF/IVC (Vit-IVM group); and group IV) IVM/IVF of vitrified oocytes followed by IVC wherein the embryos were exposed to Ang II on day 4 of IVC (Vit-D4 group). The embryos were immunostained with primary antibodies against Na+/K+/ATPase α1 and β1 subunits. Results: In Vit-IVM and Vit-D4 groups, the rates of expanded and total blastocysts on day 7 as well as the proportion of blastocysts on day 8 were increased. The expression of Na+/K+/ATPase α1 and β1 subunits were positively influenced by the addition of Ang II on day 4 (Vit-D4 group). Conclusion: The addition of Ang II to the IVM and IVC media could improve blastocysts formation in vitrified sheep oocytes. This improvement might be related to the greater expression of Na+/K+/ATPase α1 and β1 subunits when Ang II was added during IVC. 139 144 Mohammad Mehdi Naderi Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Sara Borjian Boroujeni Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Ali Sarvari Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Banafsheh Heidari 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 Amir-Hassan Zarnani Reproductive Immunology Research Center, Avicenna Research Institute, ACECR Reproductive Immunology Research Center, Avicenna Research Institute, ACECR Iran Abolfazl Shirazi Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECRResearch Institute of Animal Embryo Technology, Shahrekord University Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECRResearch Institute of Animal Embryo Technology, Shahrekord University IranIran Angiotensin IINa+/K+/ATPaseOocyteOvine Vitrification 247.pdf Luciano AM, Chigioni S, Lodde V, Franciosi F, Luvoni GC, Modina SC. 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Leptin Receptor Gene Polymorphism may Affect Subclinical Atherosclerosis in Patients with Acromegaly 2 2 Background: Acromegaly is associated with increased morbidity and mortality related to cardiovascular diseases. Leptin (LEP) and Leptin Receptor (LEPR) gene polymorphisms can increase cardiovascular risks. The aim of this study was to investigate association between the frequencies of LEP and LEPR gene polymorphisms and subclinical atherosclerosis in acromegalic patients. Methods: Forty-four acromegalic patients and 30 controls were admitted to study. The polymorphisms were identified by using polymerase chain reaction from peripheral blood samples. The levels of systolic and diastolic blood pressure, BMI, fasting plasma glucose, fasting insulin, IGF-I, GH, IGFBP3, leptin, triglyceride, carotid Intima Media Thickness (cIMT) and HDL and LDL cholesterol concentrations were evaluated. Results: There was statistically significant difference between the LEPR genotypes of acromegalic patients (GG 11.4%, GA 52.3%, and AA 36.4%) and controls (GG 33.3%, GA 50%, and AA 16.7%) although their LEP genotype distribution was similar. In addition, the prevalence of the LEPR gene G and A alleles was significantly different between patients and controls. No significant difference was found among the G(-2548)A leptin genotypes of groups in terms of the clinical parameters. cIMT significantly increased homozygote LEPR GG genotype group compared to AA subjects in patients. But the other parameters were not different between LEPR genotypes groups of patients and controls. Conclusion: It can be said that the LEPR gene polymorphism may affect cIMT in patients. The reason is that LEPR GG genotype carriers may have more risk than other genotypes in the development of subclinical atherosclerosis in acromegaly. 145 150 Sebahat Turgut Department of Physiology, Faculty of Medicine, University of Pamukkale Department of Physiology, Faculty of Medicine, University of Pamukkale Turkey Senay Topsakal Division of Endocrinology, Department of Internal Medicine, Faculty of Medicine, University of Pamukkale Division of Endocrinology, Department of Internal Medicine, Faculty of Medicine, University of Pamukkale Turkey Melek Tunç Ata Department of Physiology, Faculty of Medicine, University of Pamukkale Department of Physiology, Faculty of Medicine, University of Pamukkale Turkey Duygu Herek Department of Radiology, Faculty of Medicine, University of Pamukkale Department of Radiology, Faculty of Medicine, University of Pamukkale Turkey Fulya Akın Division of Endocrinology, Department of Internal Medicine, Faculty of Medicine, University of Pamukkale Division of Endocrinology, Department of Internal Medicine, Faculty of Medicine, University of Pamukkale Turkey Şeyma Özkan Department of Physiology, Faculty of Medicine, University of Pamukkale Department of Physiology, Faculty of Medicine, University of Pamukkale Turkey Günfer Turgut Department of Physiology, Faculty of Medicine, University of Pamukkale Department of Physiology, Faculty of Medicine, University of Pamukkale Turkey AcromegalyLeptinPolymorphism 248.pdf Colao A, Ferone D, Marzullo P, Lombardi G. 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