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Research Institutions without Walls 2 2 A network can be loosely defined as a structure linking together individual and organizational actors with shared goals or values, though often not a shared geography. A large body of literature highlights the important interaction between knowledge and networks. Interest in the impact of networking on knowledge translation and exchange, diffusion of innovations, knowledge management, and organizational outcomes is also increasing 1. There has been a growing interest in research networks and its implications on the creation of new knowledge. For example, there seems to be a consensus that those "scientists who collaborate with each other are more productive, oftentimes producing 'better' science, than are individual investigators. An open science platform can empowers researchers in their daily work and where everybody has equal opportunity to seek, share and generate knowledge. A value network can be defined as a network of relationships, which creates both tangible and intangible value through a complicated dynamic exchange between individuals, groups and organisations 2. The partnership for research and innovation in the health system funding opportunity recognizes the need to create networks of health researchers and clinical practitioners that can generate solutions to improve sustainable quality and value for money in the health system. The partnership for health research and innovation in the health system will support research and innovation. It seems that the concept of research networking in developing countries with several limitations such as research budgets should be engraved in the minds rather than papers. 63 63 Shahin Akhondzadeh Tehran University of Medical Sciences Tehran University of Medical Sciences Iran Editorial 153.pdf Abbott S, Petchey R, Kessel A, Killoran A. What sort of networks are public health networks? Public Health 2006;120(6):551-556.##Hara N, Hew KF. Knowledge sharing in an online community of health care professionals. Inform Technol People 2007;20(3):235-261.##

Computational Survey of FHIT, A Putative Human Tumor Suppressor, Truncates Structure 2 2 Background: Fragile Histidine Triad protein (FHIT), as a known tumor suppressor protein, has been proposed to play crucial role in inhibiting p53 degradation by MDM2. Studies have confirmed FHIT interaction with p53 or MDM2, although functional interacting domains of FHIT with MDM2 and/or p53 are not completely defined. Thus, through determining the significant structural interacting domains of FHIT, information with regard to MDM2 and p53 would be provided. As there were no previous studies evaluating the interaction of optimized important parts of target molecules, docking study was employed. Methods: Truncated structures of FHIT were screened to reveal critical sections engaging in FHIT interaction. HEX program was used in order to study the interaction of target structures. Results: Given the total energy, FHIT structures (β5-7, α1) and (α1) of FHIT were showed to be better candidates in comparison with other structures in interaction with optimized MDM2 part. Furthermore, FHIT structures (β4-7, α1) and (β5-7, α1) were considered to be better than other structures in interaction with optimized p53 part. FHIT truncates which interact with MDM2 optimized part exhibited lower energy levels than FHIT truncates which interact with p53 optimized part. Conclusion: Our results can be useful for designing new inhibitors of this protein complex interaction which would result in tumor repression. 64 71 Ameneh Eslamparast Biotechnology Research Center, Pasteur Institute of Iran Iran Mohammad Hossein Ghahremani Department of Pharmacology-Toxicology, Faculty of Pharmacy, Tehran University of Medical Sciences Department of Pharmacology-Toxicology, Faculty of Pharmacy, Tehran University of Medical Sciences Iran Soroush Sardari Drug Design and Bioinformatics Unit, Medical Biotechnology Department, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran Iran Fragile histidine triad proteinMDM2 proteinTumor suppressor proteins 144.pdf Croce CM, Sozzi G, Huebner K. Role of FHIT in human cancer. J Clin Oncol 1999;17(5):1618-1624.##Deng WG, Nishizaki M, Fang BL, Roth JA, Ji L. Induction of apoptosis by tumor suppressor FHIT via death receptor signaling pathway in human lung cancer cells. Biochem Biophys Res Commun 2007;355(4):993-999.##Woenckhaus M, Merk J, Stoehr R, Schaeper F, Gaumann A, Wiebe K, et al. Prognostic value of FHIT, CTNNB1, and MUC1 expression in non-small cell lung cancer. Hum Pathol 2008;39(1):126-136.##Kohno T, Yokota J. How many tumor suppressor genes are involved in human lung carcinogenesis? Carcinogenesis 1999;20(8):1403-1410.##Yang Q, Yoshimura G, Sakurai T, Kakudo K. The Fragile Histidine Triad gene and breast cancer. Med Sci Monit 2002;8(7):RA140-144.##Terry G, Ho L, Londesborough P, Duggan C, Hanby A, Cuzick J. The expression of FHIT, PCNA and EGFR in benign and malignant breast lesions. Br J Cancer 2007;96(1):110-117.##Greenspan DL, Connolly DC, Wu R, Lei RY, Vogelstein JTC, Kim YT, et al. Loss of FHIT expression in cervical carcinoma cell lines and primary tumors. Cancer Res 1997;57(21):4692-4698.##Cao J, Li W, Xie J, Du H, Tang W, Wang H, et al. Down-regulation of FHIT inhibits apoptosis of colorectal cancer: mechanism and clinical implication. Surg Oncol 2006;15(4):223-233.##Moore CD, Shahrokh K, Sontum SF, Cheatham TE, Yost GS. Improved cytochrome P450 3A4 molecular models accurately predict the Phe215 requirement for raloxifene dehydrogenation selectivity. Biochemistry 2010;49(41):9011-9019.##Ding Y, Larson G, Rivas G, Lundberg C, Geller L, Ouyang C, et al. S Strong signature of natural selection within an FHIT intron implicated in prostate cancer risk. Plos One 2008;3(10):e3533.##Ohta M, Inoue H, Cotticelli MG, Kastury K, Baffa R, Palazzo J, et al. The FHIT gene, spanning the chromosome 3p14.2 fragile site and renal carcinoma-associated t(3;8) breakpoint, is abnormal in digestive tract cancers. Cell 1996;84(4):587-597.##Baffa R, Veronese ML, Santoro R, Mandes B, Palazzo JP, Rugge M, et al. Loss of FHIT expression in gastric carcinoma. Cancer Res 1998;58(20):4708-4714.##Perez-Ordonez B, Beauchemin M, Jordan RCK. Molecular biology of squamous cell carcinoma of the head and neck. J Clin Pathol 2006;59(5):445-453.##Nishizaki M, Sasaki J, Fang B, Atkinson EN, Minna JD, Roth JA, et al. Synergistic tumor suppression by coexpression of FHIT and p53 coincides with FHIT-mediated MDM2 inactivation and p53 stabilization in human non-small cell lung cancer cells. Cancer Res 2004;64(16):5745-5752.##Orengo CA, Thornton JM. Alpha plus beta folds revisited: some favoured motifs. Structure 1993;1(2):105-120.##Lima CD, D Amico KL, Naday I, Rosenbaum G, Westbrook EM, Hendrickson WA. MAD analysis of FHIT, a putative human tumor suppressor from the HIT protein family. Structure 1997;5(6):763-774.##Freedman DA, Epstein CB, Roth JC, Levine AJ. A genetic approach to mapping the p53 binding site in the MDM2 protein. Mol Med 1997;3(4):248-259.##Freedman DA, Levine AJ. Regulation of the p53 protein by the MDM2 oncoprotein-thirty-eighth G.H.A. Clowes Memorial Award Lecture. Cancer Res 1999;59(1):1-7.##Cavazzoni A, Galetti M, Fumarola C, Alfieri RR, Roz L, Andriani F, et al. Effect of inducible FHIT and p53 expression in the Calu-1 lung cancer cell line. Cancer Lett 2007;246(1-2):69-81.##Fan H, Wang X, Zhu J, Robillard GT, Mark AE. Molecular dynamics simulations of the hydrophobin SC3 at a hydrophobic/hydrophilic interface. Proteins 2006;64(4):863-873.##Eswar N, Webb B, Marti-Renom MA, Madhusudhan MS, Eramian D, Shen MY, et al. Comparative protein structure modeling using MODELLER. Curr Protoc Protein Sci 2007;Chapter 2:Unit 2.9.##Fahham N, Ghahremani MH, Sardari S, Vaziri B, Ostad SN. Simulation of different truncated p16(INK4a) forms and in silico study of interaction with Cdk4. Cancer Inform 2009;7:1-11.##Ritchie DW, Kemp GJ. Protein docking using spherical polar fourier correlations. Proteins 2000;39(2):178-194.##Ritchie DW. Evaluation of protein docking predictions using Hex 3.1 in CAPRI rounds 1 and 2. Proteins 2003;52(1):98-106.##Oliner JD, Pietenpol JA, Thiagalingam S, Gyuris J, Kinzler KW, Vogelstein B. Oncoprotein MDM2 conceals the activation domain of tumour suppressor p53. Nature 1993;362(6423):857-860.##Chen J, Marechal V, Levine AJ. Mapping of the p53 and mdm-2 interaction domains. Mol Cell Biol 1993;13(7):4107-4114.##Lin J, Chen J, Elenbaas B, Levine AJ. Several hydrophobic amino acids in the p53 amino-terminal domain are required for transcriptional activation, binding to mdm-2 and the adenovirus 5 E1B 55-kD protein. Genes Dev 1994;8(10):1235-1246.##Zhao Y, Bernard D, Wang S. Small molecule inhibitors of MDM2-p53 and MDMX-p53 interaction as new cancer therapeutics. BioDiscovery 2013;4(8):1-15.##Chene P. Inhibition of the p53-MDM2 interaction: targeting a protein-protein interface. Mol Cancer Res 2004;2(1):20-28.##Vassilev LT, Vu BT, Graves B, Carvajal D, Podlaski F, Filipovic Z, et al. In vivo activation of the p53 pathway by small-molecule antagonists of MDM2. Science 2004;303(5659):844-848.##Fuchs SY, Adler V, Buschmann T, Wu X, Ronai Z. Mdm2 association with p53 targets its ubiquitination. Oncogene 1998;17(19):2543-2547.##Michael D, Oren M. The p53-Mdm2 module and the ubiquitin system. Semin Cancer Biol 2003;13(1):49-58.##Moll UM, Petrenko O. The MDM2-p53 interaction. Mol Cancer Res 2003;1(14):1001-1008.##Bahnassy AA, Zekri AR, Madbouly MS, El-Naggar M, El-Khelany ZF, El-Merzebany MM. The correlation between FHIT, P53 and MMR genes in human papillomavirus-associated cervical carcinoma. J Egypt Natl Canc Inst 2006;18(3):191-202.##Grasberger BL, Lu T, Schubert C, Parks DJ, Carver TE, Koblish HK, et al. Discovery and cocrystal structure of benzodiazepinedione HDM2 antagonists that activate p53 in cells. J Med Chem 2005; 48(4):909-912.##

Development of a High-resolution Melting Analysis Method Based on SYBR Green-I for rs7216389 Locus Genotyping in Asthmatic Child Patients 2 2 Background: Asthma is caused by the combination of different factors. Current concepts of asthma pathogenesis emphasize on gene-environment interactions. Mega-genome scanning projects revealed that different Single Nucleotide Polymorphisms (SNPs) are related to asthma susceptibility. rs7216389-T is one of them that is related to childhood asthma and its effect on childhood asthma severity has been proved in different nations, however no study has been performed in Eastern Mediterranean and Middle East countries yet. Methods: To perform population genetic studies, a rapid and high-throughput screening method is necessary. High-resolution melting analysis is a rapid, powerful and accurate method, which is suitable for this type of studies. Therefore, it has been decided to develop a high-resolution melting method for rs7216389 locus genotyping in Iranian asthmatic children. In the current study, a high-resolution melting analysis method based on SYBR Green-I was developed to check the frequency of rs7216389-T mutation in Iranian asthmatic children for the first time. Results: Second and third classes of intercalating dyes are commonly used for high-resolution melting method. However, in this study, SYBR Green-I was used for rs7216389 locus genotyping for the first time. Our results for 60 samples showed that SYBR Green-I has good efficacy for rs7216389 locus genotyping through high-resolution melting method in comparison with PCR-RFLP and sequencing. Conclusion: Comparison of our results based on HRM analysis with PCR-RFLP showed that our developed method is rapid, accurate, high-throughput and economic to study the rs7216389 locus in asthmatic children and it is applicable for other similar population genetic studies. 72 80 Zahra Vali Malaria and Vector Research Group (MVRG), Biotechnology Research Center, Pasteur Institute of IranDepartment of Pediatrics, Shahid Sadoughi Hospital, School of Medicine, Shahid Sadoughi University of Medical Sciences Department of Pediatrics, Shahid Sadoughi Hospital, School of Medicine, Shahid Sadoughi University of Medical Sciences IranIran Abbasali Raz Malaria and Vector Research Group (MVRG), Biotechnology Research Center, Pasteur Institute of Iran Iran Hanieh Bokharaei Malaria and Vector Research Group (MVRG), Biotechnology Research Center, Pasteur Institute of Iran Iran Mohammad Nabavi Department Allergy and Immunology, Hazrate Rasoul Akram Hospital, Tehran University of Medical Sciences Department Allergy and Immunology, Hazrate Rasoul Akram Hospital, Tehran University of Medical Sciences Iran Mohammad Hassan Bemanian Department Allergy and Immunology, Hazrate Rasoul Akram Hospital, Tehran University of Medical Sciences Department Allergy and Immunology, Hazrate Rasoul Akram Hospital, Tehran University of Medical Sciences Iran Mina Sharifi Yazdi Boo-Ali Polyclinic of Social Security Organization Iran Navid Dinparast Djadid Malaria and Vector Research Group (MVRG), Biotechnology Research Center, Pasteur Institute of Iran Iran AsthmaChildReal-time PCRSYBR Green-I 145.pdf Du R, Litonjua AA, Tantisira KG, Lasky-Su J, Sunyaev SR, Klanderman BJ, et al. Genome-wide association study reveals class I MHC-restricted T cell-associated molecule gene (CRTAM) variants interact with vitamin D levels to affect asthma exacerbations. J Allergy Clin Immunol 2012;129(2):368-373.##Duffy DL, Martin NG, Battistutta D, Hopper JL, Mathews JD. Genetics of asthma and hay fever in Australian twins. Am Rev Respir Dis 1990;142(6 Pt 1):1351-1358.##Hoffjan S, Nicolae D, Ober C. Association studies for asthma and atopic diseases: a comprehensive review of the literature. Respir Res 2003;4:14.##Zhang J, Pare PD, Sandford AJ. Recent advances in asthma genetics. Respir Res 2008;9:4.##Moffatt MF, Kabesch M, Liang L, Dixon AL, Strachan D, Heath S, et al. Genetic variants regulating ORMDL3 expression contribute to the risk of childhood asthma. Nature 2007;448(7152):470-473.##Bisgaard H, Bonnelykke K, Sleiman PM, Brasholt M, Chawes B, Kreiner-Moller E, et al. Chromosome 17q21 gene variants are associated with asthma and exacerbations but not atopy in early childhood. Am J Respir Crit Care Med 2009;179(3):179-185.##Bouzigon E, Corda E, Aschard H, Dizier MH, Boland A, Bousquet J, et al. Effect of 17q21 variants and smoking exposure in early-onset asthma. N Engl J Med 2008;359(19):1985-1994.##Cecil JE, Tavendale R, Watt P, Hetherington MM, Palmer CN. An obesity-associated FTO gene variant and increased energy intake in children. N Engl J Med 2008;359(24):2558-2566.##Halapi E, Gudbjartsson DF, Jonsdottir GM, Bjornsdottir US, Thorleifsson G, Helgadottir H, et al. A sequence variant on 17q21 is associated with age at onset and severity of asthma. Eur J Hum Genet 2010;18(8):902-908.##Hirota T, Harada M, Sakashita M, Doi S, Miyatake A, Fujita K, et al. Genetic polymorphism regulating ORM1-like 3 (Saccharomyces cerevisiae) expression is associated with childhood atopic asthma in a Japanese population. J Allergy Clin Immunol 2008;121(3):769-770.##Madore AM, Tremblay K, Hudson TJ, Laprise C. Replication of an association between 17q21 SNPs and asthma in a French-Canadian familial collection. Hum Genet 2008;123(1):93-95.##Tavendale R, Macgregor DF, Mukhopadhyay S, Palmer CN. A polymorphism controlling ORMDL3 expression is associated with asthma that is poorly controlled by current medications. J Allergy Clin Immunol 2008 ;121(4):860-863.##Sleiman PM, Annaiah K, Imielinski M, Bradfield JP, Kim CE, Frackelton EC, et al. ORMDL3 variants associated with asthma susceptibility in North Americans of European ancestry. J Allergy Clin Immunol 2008;122(6):1225-1227.##Vossen RH, Aten E, Roos A, den Dunnen JT. High-resolution melting analysis (HRMA): more than just sequence variant screening. Hum Mutat 2009;30(6):860-866.##Wittwer CT. High-resolution DNA melting analysis: advancements and limitations. Hum Mutat 2009;30(6):857-859.##Galanter J, Choudhry S, Eng C, Nazario S, Rodriguez-Santana JR, Casal J, et al. ORMDL3 gene is associated with asthma in three ethnically diverse populations. Am J Respir Crit Care Med 2008;177(11):1194-1200.##Price EP, Smith H, Huygens F, Giffard PM. High-resolution DNA melt curve analysis of the clustered, regularly interspaced short-palindromic-repeat locus of Campylobacter jejuni. Appl Environ Microbiol 2007;73(10):3431-3436.##Kagkli DM, Folloni S, Barbau-Piednoir E, Van den Eede G, Van den Bulcke M. Towards a pathogenic Escherichia coli detection platform using multiplex SYBR(R)Green Real-time PCR methods and high resolution melting analysis. PLoS One 2012;7(6):e39287.##Boulet LP, FitzGerald JM, Levy ML, Cruz AA, Pedersen S, Haahtela T, et al. A guide to the translation of the Global Initiative for Asthma (GINA) strategy into improved care. Eur Respir J 2012;39(5):1220-1229.##Jung M, Muche JM, Lukowsky A, Jung K, Loening SA. Dimethyl sulfoxide as additive in ready-to-use reaction mixtures for real-time polymerase chain reaction analysis with SYBR Green I dye. Anal Biochem 2001;289(2):292-295.##Karsai A, Muller S, Platz S, Hauser MT. Evaluation of a homemade SYBR green I reaction mixture for real-time PCR quantification of gene expression. Biotechniques 2002;32(4):790-792, 794-796.##Zipper H, Brunner H, Bernhagen J, Vitzthum F. Investigations on DNA intercalation and surface binding by SYBR Green I, its structure determination and methodological implications. Nucleic Acids Res 2004;32(12):e103.##Distefano G, Caruso M, La Malfa S, Gentile A, Wu S-B. High resolution melting analysis is a more sensitive and effective alternative to gel-based platforms in analysis of SSR--an example in citrus. PLoS One 2012;7(8):e44202.##Wittwer CT, Reed GH, Gundry CN, Vandersteen JG, Pryor RJ. High-resolution genotyping by amplicon melting analysis using LCGreen. Clin Chem 2003;49(6):853-860.##Reed GH, Wittwer CT. 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An In silico Based Comparison of Drug Interactions in Wild and Mutant Human β-tubulin through Docking Studies 2 2 Background: Tubulin protein being the fundamental unit of microtubules is actively involved in cell division thus making them a potential anti-cancer drug target. In spite of many reported drugs against tubulin, few of them have started developing resistance in human β-tubulin due to amino acid substitutions. Methods: In this study we generated three mutants (F270V, A364T and Q292E) using Modeller9v10 which were targeted with compounds from higher and lower plants along with marine isolates using iGEMDOCK2.0 to identify their residual interactions. Results: The mutant F270V does not bring in any increase in the binding affinity in comparison with the taxol-wild type due to their conservative substitutions. However, it increases the volume of the active site. A364T mutant brings a better binding among few of the marine and higher plants isolates due to the substitution of the non-reactive methyl group with the polar residue. But this leads to reduced active site volume. Finally the mutant Q292E from epothilone binding site brings a remarkable change in drug binding in the mutants in comparison with the wild type due to the substitution of uncharged residue with the charged one. But as such there was no change in the volume of the active site observed in them. Conclusion: Lower plants extracts were reported to exhibit better interactions with the taxol and epothilone binding sites. Whereas marine and higher plants isolates shows significant interactions only in the wild type instead of the mutants. In addition to this, the residual substitutions were also found to alter the conformations of the active sites in mutants. 81 93 Selvaakumar Chellasamy Department of Biotechnology and Bioinformatics, Padmashree Dr.D.Y. 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Enrichment of Undifferentiated Type A Spermatogonia from Goat Testis Using Discontinuous Percoll Density Gradient and Differential Plating 2 2 Background: The well documented source for adult multipotent stem cells is Spermatogonial Stem Cells (SSCs). They are the foundation of spermatogenesis in the testis throughout adult life by balancing self-renewal and differentiation. The aim of this study was to assess the effect of percoll density gradient and differential plating on enrichment of undifferentiated type A spermatogonia in dissociated cellular suspension of goat testes. Additionally, we evaluated the separated fractions of the gradients in percoll and samples in differential plating at different times for cell number, viability and purification rate of goat SSCs in culture. Methods: Testicular cells were successfully isolated from one month old goat testis using two-step enzymatic digestion and followed by two purification protocols, differential plating with different times of culture (3, 4, 5, and 6 hr) and discontinuous percoll density with different gradients (20, 28, 30, and 32%). The difference of percentage of undifferentiated SSCs (PGP9.5 positive) in each method was compared using ANOVA and comparison between the highest percentage of corresponding value between two methods was carried out by t-test using Sigma Stat (ver. 3.5). Results: The highest PGP9.5 (94.6±0.4) and the lowest c-Kit positive (25.1±0.7) in Percoll method was significantly (p≤0.001) achieved in 32% percoll gradient. While the corresponding rates in differential plating method for the highest PGP9.5 positive cells (81.3±1.1) and lowest c-Kit (17.1±1.4) was achieved after 5 hr culturing (p<0.001). The enrichment of undifferentiated type A spermatogonia using Percoll was more efficient than differential plating method (p<0.001). Conclusion: Percoll density gradient and differential plating were efficient and fast methods for enrichment of type A spermatogonial stem cells from goat testes. 94 103 Banafsheh Heidari Reproductive Biotechnology Research Center, Avicenna Research Institute (ACECR) Reproductive Biotechnology Research Center, Avicenna Research Institute (ACECR) Iran Minoo Gifani Immunology Research Center (IRC), Tabriz University of Medical Science Immunology Research Center (IRC), Tabriz University of Medical Science Iran Abolfazl Shirazi 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 Behzad Baradaran Immunology Research Center (IRC), Tabriz University of Medical Science Immunology Research Center (IRC), Tabriz University of Medical Science Iran Mohammad Mehdi Naderi Reproductive Biotechnology Research Center, Avicenna Research Institute (ACECR) Reproductive Biotechnology Research Center, Avicenna Research Institute (ACECR) Iran Bahareh Behzadi 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 Niknam Lakpour Nanobiotechnology Research Center, Avicenna Research Institute (ACECR) Nanobiotechnology 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 C-kitDifferential platingGoatPercoll gradientPGP 9.5Type A spermatogonia 147.pdf Hermann BP, Sukhwani M, Hansel MC, Orwig KE. 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Comparative Proteomics Study of Streptozotocin-induced Diabetic Nephropathy in Rats’ Kidneys Transfected with Adenovirus-mediated Fibromodulin Gene 2 2 Background: Transforming Growth Factor-beta (TGF-β) activation appears to be crucial for tissue injury in Diabetic Nephropathy (DN). Fibromodulin, the small leucine-rich proteoglycan, has been proposed to be the potent TGF-β modulator. In this study, the therapeutic effects of fibromodulin in the kidneys of streptozotocin (STZ)-induced diabetic rats were investigated. Methods: Diabetic rats received intraperitoneal (IP) injections of recombinant adenovirus expression vectors (RAd5) containing fibromodulin (RAd-FMOD) and were killed after 10 weeks. Proteins were isolated from the rat kidney and separated using two-dimensional gel electrophoresis. The differentially expressed proteins were analyzed using Matrix-assisted laser desorption/ ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Results: Ten spots were identified using MALDI-TOF-MS. The identified proteins were primarily responsible for cell metabolism, cytoskeleton formation, and oxidative stress. RAd-FMOD treatment markedly attenuated the albuminuria in diabetic rats. Conclusion: Taken together, these results provide a valuable clue in exploring the mechanism underlying the therapeutic effects of fibromodulin in diabetic nephropathy suggesting that it can be a potential agent in the treatment of this disease. 104 112 Akram Maleki Zanjan Metabolic Disease Research Center, Zanjan University of Medical Sciences Zanjan Metabolic Disease Research Center, Zanjan University of Medical Sciences Iran Ali Ramazani Zanjan Pharmaceutical Biotechnology Research Center, Zanjan University of Medical Sciences Zanjan Pharmaceutical Biotechnology Research Center, Zanjan University of Medical Sciences Iran Maryam Foroutan Department of Molecular Medicine & Genetics, School of Medicine, Zanjan University of Medical Sciences Department of Molecular Medicine & Genetics, School of Medicine, Zanjan University of Medical Sciences Iran Alireza Biglari Department of Molecular Medicine & Genetics, School of Medicine, Zanjan University of Medical Sciences Department of Molecular Medicine & Genetics, School of Medicine, Zanjan University of Medical Sciences Iran Parisa Ranjzad Vascular Gene Therapy Unit, Research School of Clinical & Laboratory Sciences, Manchester Academic Health Science Centre, The University of Manchester Vascular Gene Therapy Unit, Research School of Clinical & Laboratory Sciences, Manchester Academic Health Science Centre, The University of Manchester UK Ali Awsat Mellati Zanjan Metabolic Disease Research Center, Zanjan University of Medical Sciences Zanjan Metabolic Disease Research Center, Zanjan University of Medical Sciences Iran Diabetic nephropathyFibromodulinProteomics 148.pdf Manni ME, Bigagli E, Lodovici M, Zazzeri M, Raimondi L. The protective effect of losartan in the nephropathy of the diabetic rat includes the control of monoamine oxidase type A activity. Pharmacol Res 2012;65:465-471.##Schaefer L. Small leucine-rich proteoglycans in kidney disease. J Am Soc Nephrol 2011;22:1200-1207.##Schaefer L, Raslik I, Gröne HJ, Schönherr E, Macakova K, Ugorcakova J, et al. Small proteoglycans in human diabetic nephropathy: discrepancy between glomerular expression and protein accumulation of decorin, biglycan, lumican, and fibromodulin. FASEB J 2001;15(3):559-561.##Zheng Z, Nguyen C, Zhang X, Khorasani H, Wang JZ, Zara JN, et al. Delayed wound closure in fibromodulin-deficient mice is associated with increased TGF-beta3 signaling. J Invest Dermatol 2011;131(3):769-778.##Aaltonen P, Luimula P, Åström E, Palmen T, Grönholm T, Palojoki E, et al. Changes in the expression of nephrin gene and protein in experimental diabetic nephropathy. Lab Invest 2001;81(9):1185-1190.##Kritz AB, Wan S, Yim AP, Kingston PA, Baker AH. 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In vitro Therapeutic Effects of Low Level Laser at mRNA Level on the Release of Skin Growth Factors from Fibroblasts in Diabetic Mice 2 2 Background: Numerous in vitro reports suggest that Low Level Laser Therapy (LLLT) affects cellular processes by biostimulation, however most of them emphasize on using visible light lasers which have low penetration. The aim of this study was to determine the effect of infrared laser light (which is more useful in clinic because of its higher penetration) on secretion of Fibroblast Growth Factor (FGF), Platelet Derived Growth Factor (PDGF) and Vascular Endothelial Growth Factor (VEGF), as important growth factors in wound healing. Methods: Fibroblasts were extracted from the skin of 7 diabetic and 7 nondiabetic mice and cultured. Cell cultures of experimental group were irradiated with single dose of LLLT (energy density of 1 J/cm2) using an 810 nm continuous wave laser and the control group was not irradiated. Secretion of growth factors by skin fibroblasts were quantified through real time polymerase chain reaction. Results: Diabetic irradiated group showed significant increase in FGF (p=0.017) expression, although PDGF increased and VEGF decreased in both diabetic and nondiabetic irradiated groups, but these variations were not statistically significant. Conclusion: These results suggest that LLLT may play an important role in wound healing by stimulating the fibroblasts. 113 118 Nooshafarin Kazemi khoo Department of Medical Genetics, Tehran University of Medical Sciences Department of Medical Genetics, Tehran University of Medical Sciences Iran Mohammad Ali Shokrgozar National Cell Bank of Iran, Pasteur Institute of Iran Iran Iraj Ragerdi Kashani Department of Anatomy, School of Medicine, Tehran University of Medical Sciences Department of Anatomy, School of Medicine, Tehran University of Medical Sciences Iran Amir Amanzadeh National Cell Bank of Iran, Pasteur Institute of Iran Iran Ehsan Mostafavi Department of Epidemiology, Pasteur Institute of Iran Iran Hassan Sanati National Cell Bank of Iran, Pasteur Institute of Iran Iran Laleh Habibi Department of Medical Genetics, Tehran University of Medical Sciences Department of Medical Genetics, Tehran University of Medical Sciences Iran Saeid Talebi Department of Medical Genetics, Tehran University of Medical Sciences Department of Medical Genetics, Tehran University of Medical Sciences Iran Morteaz Abouzaripour Department of Anatomy, School of Medicine, Tehran University of Medical Sciences Department of Anatomy, School of Medicine, Tehran University of Medical Sciences Iran Seyed Mohammad Akrami Department of Medical Genetics, Tehran University of Medical Sciences Department of Medical Genetics, Tehran University of Medical Sciences Iran BiostimulationCell cultureLow level laser therapyTechniquesWound healing 149.pdf Pogrel MA, Chen JW, Zhang K. Effects of low-energy gallium-aluminum-arsenide laser irradiation on cultured fibroblasts and keratinocytes. Lasers Surg Med 1997;20(4):426-432.##Hallman H, Basford J, O'Brien JF, Cummins LA. Does low-energy helium-neon laser irradiation alter "in vitro" replication of human fibroblasts? Lasers Surg Med 1988;8(2):125-129.##Van Breugel HH, Bär P. Power density and exposure time of He-Ne laser irradiation are more important than total energy dose in photo-biomodulation of human fibroblasts in vitro. Lasers Surg Med 1992;12(5):528-537.##Lubart R, Wollman Y, Friedmann H, Rochkind S, Laulicht I. Effects of visible and near-infrared lasers on cell cultures. J Photochem Photobiol B 1992;12(3):305-310.##Passarella S, Casamassima E, Molinari S, Pastore D, Quagliariello E, Catalano IM, et al. Increase of proton electrochemical potential and ATP synthesis in rat liver mitochondria irradiated in vitro by helium-neon laser. FEBS Lett 1984;175(1):95-99.##Grossman N, Schneid N, Reuveni H, Halevy S, Lubart R. 780 nm low power diode laser irradiation stimulates proliferation of keratinocyte cultures: involvement of reactive oxygen species. Lasers Surg Med 1998;22(4):212-218.##Yu W, Naim J, Lanzafame R. The effects of photo-irradiation on the secretion of TGF and PDGF from fibroblasts in vitro. Lasers Surg Med Suppl 1994;6:8.##Reddy GK, Stehno-Bittel L, Enwemeka CS. Laser photostimulation accelerates wound healing in diabetic rats. Wound Repair Regen 2001;9(3):248-255.##Nemeth AJ. Lasers and wound healing. Dermatol Clin 1993;11(4):783.##Kana JS, Hutschenreiter G, Haina D, Waidelich W. Effect of low-power density laser radiation on healing of open skin wounds in rats. Arch Surg 1981;116(3):293.##Kipshidze N, Nikolaychik V, Keelan MH, Shankar LR, Khanna A, Kornowski R, et al. Low power helium: Neon laser irradiation enhances production of vascular endothelial growth factor and promotes growth of endothelial cells in vitro. Lasers Surg Med 2001;28(4):355-364.##Saygun I, Karacay S, Serdar M, Ural AU, Sencimen M, Kurtis B. Effects of laser irradiation on the release of basic fibroblast growth factor (bFGF), insulin like growth factor-1 (IGF-1), and receptor of IGF-1 (IGFBP3) from gingival fibroblasts. Lasers Med Sci 2008;23(2):211-215.##Moore P, Ridgway TD, Higbee RG, Howard EW, Lucroy MD. Effect of wavelength on low-intensity laser irradiation-stimulated cell proliferation in vitro. Lasers Surg Med 2005;36(1):8-12.##Martin G, Sprague C, Epstein C. Replicative life-span of cultivated human cells. Effects of donor's age, tissue, and genotype. Lab Invest 1970;23(1):86-92.##Goldstein S, Littlefield JW, Soeldner JS. Diabetes mellitus and aging: diminished plating efficiency of cultured human fibroblasts. Proc Natl Acad Sci USA 1969;64(1):155-160.##Loots MA, Kenter SB, Au FL, Van Galen W, Middelkoop E, Bos JD, et al. Fibroblasts derived from chronic diabetic ulcers differ in their response to stimulation with EGF, IGF-I, bFGF and PDGF-AB compared to controls. Eur J Cell Biol 2002;81(3):153-160.##Grazul-Bilska A, Luthra G, Reynolds L, Bilski J, Johnson M, Adbullah SA, et al. Effects of basic fibroblast growth factor (FGF-2) on proliferation of human skin fibroblasts in type II diabetes mellitus. Exp Clin Endocrinol Diabetes 2002;110(4):176-181.##Greenhalgh D, Sprugel K, Murray M, Ross R. PDGF and FGF stimulate wound healing in the genetically diabetic mouse. Am J Pathol 1990;136(6):1235.##Hammes H-P, Lin J, Bretzel RG, Brownlee M, Breier G. Upregulation of the vascular endothelial growth factor/vascular endothelial growth factor receptor system in experimental background diabetic retinopathy of the rat. 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How long after laser irradiation should cellular responses be measured to determine the laser effect? J Laser Appl 2007;19:74.##Pourreau-Schneider N, Ahmed A, Soudry M, Jacquemier J, Kopp F, Franquin JC, et al. Helium-neon laser treatment transforms fibroblasts into myofibroblasts. Am J Pathol 1990;137(1):171.##Hawkins D, Houreld N, Abrahamse H. Low level laser therapy (LLLT) as an effective therapeutic modality for delayed wound healing. Ann NY Acad Sci 2005;1056:486-493.##Kazemi khoo N. Successful treatment of diabetic foot ulcers with low-level laser therapy. Foot 2006;16:184-187.##Kazemi khoo N, Iravani A, Arjmand M, Vahabi F, Lajevardi M, Akrami SM, et al. A metabolomic study on the effect of intravascular laser blood irradiation on type 2 diabetic patients. Lasers Med Sci 2013;28(6):1-6.##Khamseh ME, Kazemi khoo N, Aghili R, Forough B, Lajevardi M, Hashem Dabaghian F, et al. Diabetic distal symmetric polyneuropathy: Effect of low-intensity laser therapy. 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An In vitro Study on Chick Somite Ability to Express Cerberus, Chordin, FGF8,Follistatin, and Noggin Transcripts 2 2 Background: In vitro simulation of developmental processes is an invaluable tool to shed light on the intrinsic mechanism of developmental biosystems such as central nervous system in mammals. Chick somites have been used to simulate the neural differentiation of human neural progenitor cells. In the present study, we aimed to indicate whether somites have the ability to express required neural differentiation factors at mRNA level. Methods: Chick embryos were isolated from the yolk surface of the fertilized eggs and somites were subsequently isolated from embryos under a dissecting microscope. Total RNA of the somites was extracted and RT-PCR carried out with specific primers of cerberus, chordin, FGF8, follistatin and noggin. Results: Data showed that five aforementioned factors were co-expressed after 7 days in vitro by somites. Conclusion: We concluded that neural induction property of somites appeared by production of required neural differentiation factors including cerberus, chordin, FGF8, follistatin and noggin. 119 122 Samaneh Sadat Hosseini Farahabadi Cell and Molecular Biology Division, Department of Biology, School of Sciences, University of Isfahan Cell and Molecular Biology Division, Department of Biology, School of Sciences, University of Isfahan Iran Khadijeh Karbalaie Department of Cellular Biotechnology at Cell Science Research Center, Royan Institute for Biotechnology, ACECR Department of Cellular Biotechnology at Cell Science Research Center, Royan Institute for Biotechnology, ACECR Iran Hossein Salehi Department of Anatomy, School of Medicine, Isfahan University of Medical Sciences Department of Anatomy, School of Medicine, Isfahan University of Medical Sciences Iran Farzaneh Rabiee Department of Cellular Biotechnology at Cell Science Research Center, Royan Institute for Biotechnology, ACECR Department of Cellular Biotechnology at Cell Science Research Center, Royan Institute for Biotechnology, ACECR Iran Kamran Ghaedi Cell and Molecular Biology Division, Department of Biology, School of Sciences, University of IsfahanDepartment of Cellular Biotechnology at Cell Science Research Center, Royan Institute for Biotechnology, ACECR Cell and Molecular Biology Division, Department of Biology, School of Sciences, University of IsfahanDepartment of Cellular Biotechnology at Cell Science Research Center, Royan Institute for Biotechnology, ACECR IranIran Mohammad-Hossein Nasr-Esfahani Department of Cellular Biotechnology at Cell Science Research Center, Royan Institute for Biotechnology, ACECR Department of Cellular Biotechnology at Cell Science Research Center, Royan Institute for Biotechnology, ACECR Iran Cerberus proteinChordinFGF8 proteinFollistatinNoggin proteinSomites 150.pdf Gilbert SF. Developmental biology. 9th ed. Sunderland, Mass: Sinauer associates; 2010.##Rowan AM, Stern CD, Storey KG. Axial mesendoderm refines rostrocaudal pattern in the chick nervous system. Development 1999;126(13):2921-2934. ##Salehi H, Karbalaie K, Salamian A, Kiani A, Razavi S, Nasr-Esfahani MH, et al. Differentiation of human ES cell-derived neural progenitors to neuronal cells with regional specific identity by co-culturing of notochord and somite. Stem Cell Res 2012;8(1):120-133. ##Hamburger V, Hamilton HL. A series of normal stages in the development of the chick embryo. J Morphol 1951;88(1):49-92.##Sagha M, Karbalaie K, Tanhaee S, Esfandiari E, Salehi H, Sadeghi-Aliabadi H, et al. Neural induction in mouse embryonic stem cells by co-culturing with chicken somites. Stem Cells Dev 2009;18(9):1351-1360.##Wilson L, Maden M. The mechanisms of dorsoventral patterning in vertebrate neural tube. Dev Biol 2005;282(1):1-13.##Lim DA, Tramontin AD, Trevejo JM, Herrera DG, García-Verdugo JM, Alvarez-Buylla A. Noggin antagonizes BMP signaling to create a niche for adult neurogenesis. Neuron 2000;28(3):713-726.##Sasai Y, Lu B, Steinbeisser H, De Robertis EM. Regulation of neural induction by the Chd and Bmp-4 antagonistic patterning signals in Xenopus. Nature 1995;376(6538):333-336.##Urshansky N, Mausner-Fainberg K, Auriel E, Regev K, Karni A. Low and dysregulated production of follistatin in immune cells of relapsing-remitting multiple sclerosis patients. J Neuroimmunol 2011;238(1-2):96-103.##Yu X, He F, Zhang T, Espinoza-Lewis RA, Lin L, Yang J, et al. Cerberus functions as a BMP agonist to synergistically induce nodal expression during left-right axis determination in the chick embryo. Dev Dyn 2008;237(12):3613-3623.##Lahti L, Saarimäki-Vire J, Rita H, Partanen J. FGF signaling gradient maintains symmetrical proliferative divisions of midbrain neuronal progenitors. 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The Synergic Effects of Crocus Sativus L. and Low Frequency Electromagnetic Field on VEGFR2 Gene Expression in Human Breast Cancer Cells 2 2 Background: Angiogenesis, which is required for embryonic development and many physiological events, plays crucial role in many pathological conditions such as tumor growth and metastasis. Recent studies indicate anticancer and antitumor properties of saffron against human cancers. Many processes are affected by Electromagnetic Field (EMF) and its effect on proliferation and gene expression were examined. In this experimental study, the synergic effects of saffron and EMF on VEGFR2 gene expression in MCF7 cells were investigated. Methods: Saffron was extracted using freeze dryer. MCF7 cells were grown in RPMI1640 medium supplemented with 10% FBS and incubated at 37C with 5% CO2. After 24 hr cells were treated with saffron extract at concentrations of 100, 200, 400 and 800 μg/ml. Forty eight hr after treatment all flasks were exposed with EMF (50 Hz, 0.004 T). Then total RNA was extracted and cDNA was synthetized using specific primer. Synthetized products were analyzed by Real Time PCR to determine expression level of VEGFR2. Data were analyzed by SPSS (ANOVA & Tukey). Results: Critical inhibitory effect on VEGFR2 gene expression was 20% at 400 μg/ml. Synergic use of EMF and saffron extract showed most reduction (38%) at 100 μg/ml. On the other hand synergic use of 200, 400 and 800 μg/ml saffron aqua extract and EMF decline noticeably the VEGFR2 level of gene expression to 29, 35 and 36%, respectively. EMF itself also reduced VEGFR2 up to 25% in comparison with control group which is remarkable at p<0.001. Conclusion: Results indicate a decrease in the expression of vascular endothelial growth factor receptor in the treated samples with saffron extract compared to control. This reduction in VEGFR2 level induced by synergic treatment of saffron and EMF which reveals induction of inhibitory effects of saffron on angiogenesis and could be also considered as a promising chemotherapeutic agent in breast cancer treatment. 123 127 Marzieh Mousavi Research Center for Animal Development Applied Biology, Mashhad Branch, Islamic Azad University Research Center for Animal Development Applied Biology, Mashhad Branch, Islamic Azad University Iran Elaheh Amini Research Center for Animal Development Applied Biology, Mashhad Branch, Islamic Azad UniversityDepartment of Biology, School of Sciences, Mashhad Branch, Islamic Azad University Research Center for Animal Development Applied Biology, Mashhad Branch, Islamic Azad UniversityDepartment of Biology, School of Sciences, Mashhad Branch, Islamic Azad University IranIran Khadijeh Shahrokhabadi Department of Biology, School of Sciences, Mashhad Branch, Islamic Azad University Department of Biology, School of Sciences, Mashhad Branch, Islamic Azad University Iran AngiogenesisCancerMCF7 cellsSaffronVEGFR2 151.pdf Bhat TA, Singh RP. 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The carotenoid crocetin enhances pulmonary oxygenation. J Appl Physiol 1988;65(2):683-686.##Feizzadeh B, Afshari JT, Rakhshandeh H, Rahimi A, Brook A, Doosti H. Cytotoxic effect of saffron stigma aqueous extract on human transitional cell carcinoma and mouse fibroblast. Urol J 2008;5(3):161-167. ##Mousavi SH, Tavakkol-Afshari J, Brook A, Jafari-Anarkooli I. Role of caspases and Bax protein in saffron-induced apoptosis in MCF-7 cells. Food Chem Toxicol 2009;47(8):1909-1913.##Tavakkol-Afshari J, Brook A, Mousavi SH. Study of cytotoxic and apoptogenic properties of saffron extract in human cancer cell lines. Food Chem Toxicol 2008;46(11):3443-3447.##McKay JC, Prato FS, Thomas AW. A literature review: the effects of magnetic field exposure on blood flow and blood vessels in the microvasculature. Bioelectromagnetics 2007;28(2):81-98.##Delle Monache S, Alessandro R, Iorio R, Gualtieri G, Colonna R. Extremely low frequency electromagnetic fields (ELF-EMFs) induce in vitro angiogenesis process in human endothelial cells. Bioelectromagnetics 2008;29(8):640-648.##Chen X, Zhuang J, Kolb JF, Schoenbach KH, Beebe SJ. Long term survival of mice with hepato cellular carcinoma after pulse power ablation with nanosecond pulsed electric fields. Technol Cancer Res Treat 2012;11(1):83-93.##Wang Z, Yang P, Xu H, Qian A, Hu L, Shang P. Inhibitory effects of a gradient static magnetic field on normal angiogenesis. Bioelectromagnetics 2009;30(6):446-453.##