Autistic Disorders and Medical Biotechnology 2 2 Autistic Disorders (ADs) are neuro-developmental disorders in the category of pervasive developmental disorders chiefly described by three main deficits: 1) deviant communication, 2) impaired reciprocal social interaction, and 3) limited, repetitive and stereotypic patterns of behaviors or interests 1. The world-wide prevalence of ADs is estimated to be 62/10,000 1. Although various treatment regimens have been proposed for improving overall function of autistic patients, a definite efficient treatment which can target both core and associated symptoms in ADs has not been established so far. For example, current approved drugs by the Food and Drug Administration (FDA) such as risperidone and aripiprazole have not been proven to pose significant effect on the core features of this disorder 2-4. While the absolute pathophysiologic mechanism of ADs is still debated, several genetic, environmental and neurobiological factors such as immune dysfunction, oxidative stress and imbalance of the inhibitory-excitatory systems are implicated in the pathogenesis of these disorders 5-7. Neurobiological models have become research areas of interest for development of novel therapeutic agents in this regard 1. Increased neuronal excitation in various central pathways has been proposed as one of the main neurobiological dysregulations in autistic patients 1. Indeed, biotechnology and in particular gene therapies plays an important role in the future of research in autism 1. Neurexin 1: Part of family of genes that play a role with the neurotransmitter glutamate which is linked to autism. Gene chip technology was used to scan for genetic similarities in people with autism. DNA was scanned to search for copy number variations (CNVs), or insertions and deletions of genetic material linked to autism 1. Adult Form of Fragile X Syndrome: Expression of toxic RNA leads to Fragile X Tremor Ataxia Syndrome is modifiable by gene therapy 1. Fragile X Syndrome: Caused by loss of a gene called FMPR which acts as a break on a protein synthesis in specific area of the brain. This allows another protein, mGluR5 Metabotropic Glutamate Receptor, to function unchecked resulting in over activity in the brain. Reducing mGluR5 reduces symptoms associated with fragile x syndrome 1. MECP2 Activation of the gene reversed Rett syndrome. The MECP2 gene mutation is present in 95 percent of individuals with the disease 1. 1 1 Shahin Akhondzadeh Tehran University of Medical Sciences Tehran University of Medical Sciences Iran Autistic disorderBiotechnologyGenetic diseases 193.pdf Mohammadi MR, Akhondzadeh S. Autism spectrum disorders: etiology and pharmacotherapy. Current Drug Therapy 2007;2:97-103.##Akhondzadeh S, Fallah J, Mohammadi MR, Imani R, Mohammadi M, Salehi B, et al. Double-blind placebo-controlled trial of pentoxifylline added to risperidone: effects on aberrant behavior in children with autism. Prog Neuropsychopharmacol Biol Psychiatry 2010;34:32-36.##Ghaleiha A, Asadabadi M, Mohammadi MR, Shahei M, Tabrizi M, Hajiaghaee R, et al. Memantine as adjunctive treatment to risperidone in children with autistic disorder: A randomized, double-blind, placebo-controlled trial. Int J Neuropsychopharmacol 2013;16(4):783-789.##Ghaleiha A, Ghyasvand M, Mohammadi MR, Farokhnia M, Yadegari N, Tabrizi M, et al. Galantamine efficacy and tolerability as an augmentative therapy in autistic children: A randomized, double-blind, placebo-controlled trial. J Psychopharmacol 2013;28(7):677-685.##Ghaleiha A, Mohammadi E, Mohammadi MR, Farokhnia M, Modabbernia A, Yekehtaz H, et al. Riluzole as an adjunctive therapy to risperidone for the treatment of irritability in children with autistic disorder: A double-blind, placebo-controlled, randomized trial. Ped Drugs 2013;15:505-514.##Mohammadi MR, Yadegari N, Hassanzadeh E, Farokhnia M, Yekehtaz H, Mirshafiee O, et al. Double-blind, placebo-controlled trial of risperidone plus amantadine in children with autism: a 10-week randomized study. Clin Neuropharmacol 2013;36:179-184.##Asadabadi M, Mohammadi MR, Ghanizadeh A, Modabbernia A, Ashrafi M, Hassanzadeh E, et al. Celecoxib as adjunctive treatment to risperidone in children with autistic disorder: a randomized, double-blind, placebo-controlled trial. Psychopharmacology (Berl) 2013;225:51-59. ##

Production and Characterization of Monoclonal Antibodies against Human Prostate Specific Antigen 2 2 Background: Prostate Specific Antigen (PSA) is an important laboratory marker for diagnosis of prostatic cancer. Thus, development of diagnostic tools specific for PSA plays an important role in screening, monitoring and early diagnosis of prostate cancer. In this paper, the production and characterization of a panel of murine monoclonal antibodies (mAbs) against PSA have been presented.  Methods: Balb/c mice were immunized with PSA, which was purified from seminal plasma. Splenocytes of hyperimmunized mice were extracted and fused with Sp2/0 cells. By adding selective HAT medium, hybridoma cells were established and positive clones were selected by ELISA after four times of cloning. The isotypes of produced mAbs were determined by ELISA and then purified from ascitic fluids using Hi-Trap protein G column. The reactivities of the mAbs were examined with the purified PSA and seminal plasma by ELISA and western blot techniques. Furthermore, the reactivities of the mAbs were assessed in Prostate Cancer (PCa), Benign Prostatic Hyperplasia (BPH) and brain cancer tissues by Immunohistochemistry (IHC). Results: Five anti-PSA mAbs (clones: 2G2-B2, 2F9-F4, 2D6-E8, IgG1/К) and clones (2C8-E9, 2G3-E2, IgG2a/К) were produced and characterized. All mAbs, except 2F9-F4 detected the expression of PSA in PCa and BPH tissues and none of them reacted with PSA in brain cancer tissue in IHC. Besides, all mAbs could detect a protein band around 33 kDa in human seminal plasma in western blot. Conclusion: These mAbs can specifically recognize PSA and may serve as a component of PSA diagnostic kit in various biological fluids. 2 7 Ali Ahmad Bayat 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, ACECRDepartment of Molecular Medicine, School of Advanced Medical Technologies, Tehran University of Medical Sciences Monoclonal Antibody Research Center, Avicenna Research Institute, ACECRDepartment of Molecular Medicine, School of Advanced Medical Technologies, Tehran University of Medical Sciences IranIran Mahdi Shabani 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 Omid Yeganeh Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Iran Hadi Hassannia Department of Immunology, School of Public Health, Tehran University of Medical Sciences Department of Immunology, School of Public Health, Tehran University of Medical Sciences Iran Ali Sadeghitabar Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Iran Leila Balay-Goli Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Iran Farzaneh Noutash-Haghighat Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Iran Ali reza Sarrafzadeh Department of Pathology, Khatam Al Anbia Hospital Iran Mahmood Jeddi-Tehrani Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Iran ELISAImmunohistochemistryMonoclonal antibodyProstate specific antigenWestern blot 194.pdf Stat bite: Estimated worldwide cancer mortality among men, 2002. J Natl Cancer Inst 2005;97(19):1402.##Wang MC, Papsidero LD, Kuriyama M, Valenzuela LA, Murphy GP, Chu TM. Prostate antigen: a new potential marker for prostatic cancer. Prostate 1981;2(1):89-96.##Michel S, Deleage G, Charrier JP, Passagot J, Battail-Poirot N, Sibai G, et al. Anti-free prostate-specific antigen monoclonal antibody epitopes defined by mimotopes and molecular modeling. Clin Chem 1999;45(5):638-650.##Sensabaugh GF. Isolation and characterization of a semen-specific protein from human seminal plasma: a potential new marker for semen identification. J Forensic SciJ Forensic Sci 1978;23(1):106-115.##Lilja H. Structure, function, and regulation of the enzyme activity of prostate-specific antigen. World J UrolWorld J Urol 1993;11(4):188-191.##Stamey TA, Yang N, Hay AR, McNeal JE, Freiha FS, Redwine E. Prostate-specific antigen as a serum marker for adenocarcinoma of the prostate. N Engl J MedN Engl J Med 1987;317(15):909-916.##Nelson MA, Porterfield BW, Jacobs ET, Clark LC. Selenium and prostate cancer prevention. Semin Urol Oncol 1999;17(2):91-96.##Leinonen J, Niemela P, Lovgren J, Bocchi L, Pettersson K, Nevanlinna H, et al. Characterization of monoclonal antibodies against prostate specific antigen produced by genetic immunization. J Immunol MethodsJ Immunol Methods 2004;289(1-2):157-167.##Christensson A, Laurell CB, Lilja H. Enzymatic activity of prostate-specific antigen and its reactions with extracellular serine proteinase inhibitors. Eur J Biochem 1990;194(3):755-763.##Vegvari A, Rezeli M, Sihlbom C, Hakkinen J, Carlsohn E, Malm J, et al. Molecular microheterogeneity of prostate specific antigen in seminal fluid by mass spectrometry. Clin BiochemClin Biochem 2012;45(4-5):331-338.##Zhang WM, Leinonen J, Kalkkinen N, Dowell B, Stenman UH. Purification and characterization of different molecular forms of prostate-specific antigen in human seminal fluid. Clin Chem 1995;41(11):1567-1573.##Lwaleed BA, Greenfield R, Stewart A, Birch B, Cooper AJ. Seminal clotting and fibrinolytic balance: a possible physiological role in the male reproductive system. Thromb HaemostThromb Haemost 2004;92(4):752-766.##Christensson A, Lilja H. Complex formation between protein C inhibitor and prostate-specific antigen in vitro and in human semen. Eur J Biochem 1994;220(1):45-53.##Ruckle HC, Klee GG, Oesterling JE. Prostate-specific antigen: concepts for staging prostate cancer and monitoring response to therapy. Mayo Clin Proc 1994;69(1):69-79.##Bayat AA, Yeganeh O, Ghods R, Zarnani AH, Ardekani RB, Mahmoudi AR, Mahmoudian J, et al. Production and characterization of a murine monoclonal antibody against human ferritin. Avicenna J Med Biotechnol 2013;5(4):212-219.##Hadavi R, Zarnani AH, Ahmadvand N, Mahmoudi AR, Bayat AA, Mahmoudian J, et al. Production of monoclonal antibody against human nestin. Avicenna J Med Biotechnol 2010;2(2):69-77.##Sarial S, Asadi F, Jeddi-Tehrani M, Hadavi R, Bayat AA, Mahmoudian J, et al. 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Comparative Analysis of Prostate Cancer Gene Regulatory Networks via Hub Type Variation 2 2 Background: Prostate cancer is one of the most widespread cancers in men and is fundamentally a genetic disease. Identifying regulators in cancer using novel systems biology approaches will potentially lead to new insight into this disease. It was sought to address this by inferring gene regulatory networks (GRNs). Moreover, dynamical analysis of GRNs can explain how regulators change among different conditions, such as cancer subtypes. Methods: In our approach, independent gene regulatory networks from each prostate state were reconstructed using one of the current state-of-art reverse engineering approaches. Next, crucial genes involved in this cancer were highlighted by analyzing each network individually and also in comparison with each other. Results: In this paper, a novel network-based approach was introduced to find critical transcription factors involved in prostate cancer. The results led to detection of 38 essential transcription factors based on hub type variation. Additionally, experimental evidence was found for 29 of them as well as 9 new transcription factors. Conclusion: The results showed that dynamical analysis of biological networks may provide useful information to gain better understanding of the cell. 8 15 Pegah Khosravi Department of Bioinformatics, Institute of Biochemistry and Biophysics (IBB), University of TehranSchool of Biological Sciences, Institute for Research in Fundamental Sciences (IPM) Department of Bioinformatics, Institute of Biochemistry and Biophysics (IBB), University of TehranSchool of Biological Sciences, Institute for Research in Fundamental Sciences (IPM) IranIran Vahid H. Gazestani Institute of Parasitology, McGill University, Montreal Institute of Parasitology, McGill University, Montreal Canada Mohammad Akbarzadeh Department of Bioinformatics, Institute of Biochemistry and Biophysics (IBB), University of Tehran Department of Bioinformatics, Institute of Biochemistry and Biophysics (IBB), University of Tehran Iran Samira Mirkhalaf Department of Bioinformatics, Institute of Biochemistry and Biophysics (IBB), University of Tehran Department of Bioinformatics, Institute of Biochemistry and Biophysics (IBB), University of Tehran Iran Mehdi Sadeghi School of Biological Sciences, Institute for Research in Fundamental Sciences (IPM)National Institute of Genetic Engineering and Biotechnology (NIGEB) School of Biological Sciences, Institute for Research in Fundamental Sciences (IPM)National Institute of Genetic Engineering and Biotechnology (NIGEB) IranIran Bahram Goliaei Department of Bioinformatics, Institute of Biochemistry and Biophysics (IBB), University of Tehran Department of Bioinformatics, Institute of Biochemistry and Biophysics (IBB), University of Tehran Iran Gene regulatory networksProstate cancerTranscription factors 195.pdf Vogelstein B, Kinzler KW. 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Effects of Combined Soy Isoflavone Extract and Docetaxel Treatment on Murine 4T1 Breast Tumor Model 2 2 Background: Emergence of drug resistance has brought major problems in chemotherapy. Using nutrients in combination with chemotherapy could be beneficial for improvement of sensitivity of tumors to drug resistance. Soybean-derived isoflavones have been suggested as chemopreventive agents for certain types of cancer, particularly breast cancer. In this study, the synergistic effects of soy isoflavone extract in combination with docetaxel in murine 4T1 breast tumor model were investigated. Methods: In this study, mice were divided into 4 groups (15 mice per group) of control, the dietary Soy Isoflavone Extract (SIE, 100 mg/kg diet), the Docetaxel (DOCE, 10 mg/kg) injection and the combination of dietary soy isoflavone extract and intravenous docetaxel injection (DOCE+SIE). After 3 injections of docetaxel (once a week), 7 mice were sacrificed to analyze MKI67 gene and protein expressions and the rest were monitored for diet consumption, tumor growth and survival rates. Results: In DOCE+SIE group, diet consumption was significantly higher than DOCE group. While lifespan showed a trend towards improvement in DOCE+SIE group, no significant difference was observed among the 4 studied groups. Tumor volume was not significantly affected in treated groups. A lower but not significant MKI67 protein expression was detected in western blot in DOCE+SIE group. The mRNA expression was not significantly different among groups. Conclusion: The results suggest that the combination of soy isoflavone as an adjunct to docetaxel chemotherapy can be effective in improving diet consumption in breast cancer. 16 21 Ehsan Hejazi Department of Clinical Nutrition and Dietetics, School of Nutrition Sciences and Food Technology, Shahid Beheshti University of Medical Sciences Department of Clinical Nutrition and Dietetics, School of Nutrition Sciences and Food Technology, Shahid Beheshti University of Medical Sciences Iran Javad Nasrollahzadeh Department of Clinical Nutrition and Dietetics, School of Nutrition Sciences and Food Technology, Shahid Beheshti University of Medical Sciences Department of Clinical Nutrition and Dietetics, School of Nutrition Sciences and Food Technology, Shahid Beheshti University of Medical Sciences Iran Ramina Fatemi Reproductive Immunology Research Center, Avicenna Research Institute, ACECR Reproductive Immunology Research Center, Avicenna Research Institute, ACECR Iran Leila Barzegar-Yar MohamadI Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Iran Kioomars Saliminejad Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Zohre Amiri Department of Basic Sciences and Cellular and Molecular Nutrition, School of Nutrition Sciences and Food Technology, Shahid Beheshti University of Medical Sciences Department of Basic Sciences and Cellular and Molecular Nutrition, School of Nutrition Sciences and Food Technology, Shahid Beheshti University of Medical Sciences Iran Masoud Kimiagar Department of Clinical Nutrition and Dietetics, School of Nutrition Sciences and Food Technology, Shahid Beheshti University of Medical Sciences Department of Clinical Nutrition and Dietetics, School of Nutrition Sciences and Food Technology, Shahid Beheshti University of Medical Sciences Iran Mohammad Houshyari Department of Radiation Oncology, Shohada Tajrish Hospital, Shahid Beheshti University of Medical Science Department of Radiation Oncology, Shohada Tajrish Hospital, Shahid Beheshti University of Medical Science Iran Maryam Tavakoli Reproductive Immunology Research Center, Avicenna Research Institute, ACECR Reproductive Immunology Research Center, Avicenna Research Institute, ACECR Iran Farah Idali Reproductive Immunology Research Center, Avicenna Research Institute, ACECR Reproductive Immunology Research Center, Avicenna Research Institute, ACECR Iran Breast cancerDocetaxelSoy isoflavone extract 196.pdf Farinotti M, Vacchi L, Simi S, Di Pietrantonj C, Brait L, Filippini G. 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Intravenous Transplantation of Very Small Embryonic Like Stem Cells in Treatment of Diabetes Mellitus 2 2 Background: Diabetes Mellitus (DM), simply known as diabetes, refers to a group of metabolic diseases in which there are high blood sugar levels over a prolonged period. In this study, the feasibility and safety of intravenous transplantation of Very Small Embryonic Like stem cells (VSELs) were investigated for diabetes repair, and finally the migration and distribution of these cells in hosts were observed. Methods: Mouse bone marrow VSELs were isolated by Fluorescent Activating Cell Sorting (FACS) method by using fluorescent antibodies against CD45, CXCR4 and Sca1 markers. Sorted cells were analyzed for expression of oct4 and SSEA1 markers with immunocytochemistry staining method. To determine multilineage differentiation, sorted cells were differentiated to Schwann, osteocyte and beta cells. Ten days after the establishment of a mouse model of pancreas necrosis, DiI-labeled VSELs were injected into these mice via tail vein. Pancreases were harvested 4 weeks after transplantation and the sections of these tissues were observed under fluorescent microscope. Results: It was proved that CD45-, CXCR4+, and Sca1+ sorted cells express oct4 and SSEA1. Our results revealed that intravenously implanted VSELs could migrate into the pancreas of hosts and survive in the diabetic pancreas. In treated groups, blood glucose decreased significantly for at least two month and the weights of mice increased gradually.   <div> Conclusion: This study provides a strategy for using VSELs for curing diabetes and other regenerative diseases, and the strategy is considered an alternative for other stem cell types. </div> 22 31 Morteaz Abouzaripour Faculty of Medicine, Tehran University of Medical Sciences Faculty of Medicine, Tehran University of Medical Sciences Iran Iraj Ragerdi Kashani Faculty of Medicine, Tehran University of Medical Sciences Faculty of Medicine, Tehran University of Medical Sciences Iran Parichehr Pasbakhsh Faculty of Medicine, Tehran University of Medical Sciences Faculty of Medicine, Tehran University of Medical Sciences Iran Nader Atlasy Faculty of Medicine, Tehran University of Medical Sciences Faculty of Medicine, Tehran University of Medical Sciences Iran Diabetes mellitusTransplantationVery small embryonic like stem cells 197.pdf Liew CG, Andrews PW. Stem cell therapy to treat diabetes mellitus. Rev Diabet Stud 2008;5(4):203-219.##Phuc Pham Van (2011). Stem Cell Therapy for Islet Regeneration, Stem Cells in Clinic and Research, Dr. Ali Gholamrezanezhad (Ed.), ISBN: 978-953-307-797-0, InTech, Available from: http://www.intechopen.com/books/stem-cells-in-clinic-and-research/stem-cell-therapy-for-islet-regenerationwww.intechopen. ##Hussain MA, Theise ND. Stem-cell therapy for diabetes mellitus. Lancet 2004;364(9429):203-205.##Kucia M, Reca R, Campbell FR, Zuba-Surma E, Majka M, Ratajczak J, et al. A population of very small embryonic-like (VSEL) CXCR4+ SSEA-1+ Oct-4 & plus; stem cells identified in adult bone marrow. Leukemia 2006;20(5):857-869.##Wojakowski W, Tendera M, Michałowska A, Majka M, Kucia M, Maślankiewicz K, et al. Mobilization of CD34/ CXCR4+, CD34/CD117+, c-met+ stem cells, and mononuclear cells expressing early cardiac, muscle, and endothelial markers into peripheral blood in patients with acute myocardial infarction. Circulation 2004;110(20):3213-3220.##Kucia M, Dawn B, Hunt G, Guo Y, Wysoczynski M, Majka M, et al. Cells expressing early cardiac markers reside in the bone marrow and are mobilized into the peripheral blood after myocardial infarction. Circ Res 2004;95(12):1191-1199.##Paczkowska E, Larysz B, Rzeuski R, Karbicka A, Jałowiński R, Kornacewicz-Jach Z, et al. Human hematopoietic stem/progenitor-enriched CD34 (+) cells are mobilized into peripheral blood during stress related to ischemic stroke or acute myocardial infarction. Eur J Haematol 2005;75(6):461-467.##Kucia M, Zhang YP, Reca R, Wysoczynski M, Machalinski B, Majka M, et al. Cells enriches in markers of neural tissue-committed stem cells reside in the bone marrow and are mobilized into the peripheral blood following stroke. Leukemia 2006;20(1):18-28.##Halasa M, Baskiewicz-Masiuk M, Dabkowska E, Machalinski B. An efficient two-step method to purify very small embryonic-like (VSEL) stem cells from umbilical cord blood (UCB). Folia Histochemica Cytobiologica 2008;46.(2):239-238.##Ratajczak MZ, Zuba-Surma EK, Machalinski B, Ratajczak J, Kucia M. Very small embryonic-like (VSEL) stem cells: purification from adult organs, characterization, and biological significance. Stem Cell Rev 2008;4(2):89-99.##Ceradini DJ, Kulkarni AR, Callaghan MJ, Tepper OM, Bastidas N, Kleinman ME, et al. Progenitor cell trafficking is regulated by hypoxic gradients through HIF-1 induction of SDF-1. Nat Med 2004;10(8):858-864.##Ponomaryov T, Peled A, Petit I, Taichman RS, Habler L, Sandbank J, et al. Induction of the chemokine stromal-derived factor-1 following DNA damage improves human stem cell function. J Clin Invest 2000;106(11):1331-1339.##Takahashi T, Kalka C, Masuda H, Chen D, Silver M, Kearney M, et al. Ischemia- and cytokine-induced mobilization of bone marrow-derived endothelial progenitor cells for neovascularization. Nat Med 1999;5(4):434-438.##Tögel F, Isaac J, Hu Z, Weiss K, Westenfelder C. Renal SDF-1 signals mobilization and homing of CXCR4-positive cells to the kidney after ischemic injury. Kidney Int 2005;67(5):1772-1784.##Shin DM, Zuba-Surma EK, Wu W, Ratajczak J, Wysoczynski M, Ratajczak MZ, et al. Novel epigenetic mechanisms that control pluripotency and quiescence of adult bone marrow-derived Oct 4 & plus; very small embryonic-like stem cells. Leukemia 2009;23(11):2042-2051.##Annaloro C, Onida F, Lambertenghi Deliliers G. Autologous hematopoietic stem cell transplantation in autoimmune diseases. Expert Rev Hematol 2009;2(6):699-715.##Rosato E, Pisarri S, Salsano F. Current strategies for the treatment of autoimmune diseases. J Biol Regul Homeost Agents 2010;24(3):251-259.##Karussis D, Karageorgiou C, Vaknin-Dembinsky A, Gowda-Kurkalli B, Gomori JM, Kassis I, et al. Safety and immunological effects of mesenchymal stem cell transplantation in patients with multiple sclerosis and amyotrophic lateral sclerosis. Arch Neurol 2010;67(10):1187-1194.##Szodoray P, Varoczy L, Szegedi G, Zeher M. Autologous stem cell transplantation in autoimmune and rheumatic diseases: from the molecular background to clinical applications. Scand J Rheumatol 2010;39(1):1-12.##Nagaya N, Fujii T, Iwase T, Ohgushi H, Itoh T, Uematsu M, et al. Intravenous administration of mesenchymal stem cells improves cardiac function in rats with acute myocardial infarction through angiogenesis and myogenesis. Am J Physiol Heart Circ Physiol 2004;287(6):H2670-2676.##Mazo M, Planat-benard V, Abizanda G, Pelacho B, Léobon B, Gavira JJ, et al. Transplantation of adipose derived stromal cells is associated with functional improvement in a rat model of chronic myocardial infarction. Eur J Heart Fail 2008;10(5):454-462.##Mazo M, Gavira JJ, Abizanda G, Moreno C, Ecay M, Soriano M, et al. Transplantation of mesenchymal stem cells exerts a greater long-term effect than bone marrow mononuclear cells in a chronic myocardial infarction model in rat. Cell Transplant 2010;19(3):313-328.##Mark AL, Sun Z, Warren DS, Lonze BE, Knabel MK, Melville Williams GM, et al. Stem cell mobilization is life saving in an animal model of acute liver failure. Ann Surg 2010;252(4):591-596.##Shi XL, Gu JY, Han B, Xu HY, Fang L, Ding YT. magnetically labeled mesenchymal stem cells after autologous transplantation into acutely injured liver. World J Gastroenterol 2010;16(29):3674-3679.##Walker PA, Shah SK, Jimenez F, Gerber MH, Xue H, Cutrone R, et al. Intravenous multipotent adult progenitor cell therapy for traumatic brain injury: preserving the blood brain barrier via an interaction with splenocytes. Exp Neurol 2010;225(2):341-352.##Zhang H, Liu Z, Li R, Wang D, Liu W, Li J, et al. Transplantation of embryonic small hepatocytes induces regeneration of injured liver in adult rat. Transplant Proc 2009;41(9):3887-3892.##Li ZH, Liao W, Cui XL, Zhao Q, Liu M, Chen YH, et al. Intravenous transplantation of allogeneic bone marrow mesenchymal stem cells and its directional migration to the necrotic femoral head. Int J Med Sci 2011;8(1):74-83.##Sordi V, Malosio ML, Marchesi F, Mercalli A, Melzi R, Giordano T, et al. Bone marrow mesenchymal stem cells express a restricted set of functionally active chemokine receptors capable of promoting migration to pancreatic islets. Blood 2005;106(2):419-427. ##Gao F, Wu DQ, Hu YH, Jin GX. Extracellular matrix gel is necessary for in vitro cultivation of insulin producing cells from human umbilical cord blood derived mesenchymal stem cells. Chin Med J (Engl) 2008;121(9):811-818. ##Parekh VS, Joglekar MV, Hardikar AA. Differentiation of human umbilical cord blood-derived mononuclear cells to endocrine pancreatic lineage. Differentiation 2009;78(4):232-240.##Wang HS, Shyu JF, Shen WS, Hsu HC, Chi TC, Chen CP, et al. Transplantation of insulin-producing cells derived from umbilical cord stromal mesenchymal stem cells to treat NOD mice. Cell Transplant 2011;20(3):455-466.##Phuc PV, Nhung TH, Loan DT, Chung DC, Ngoc PK. Differentiating of banked human umbilical cord blood-derived mesenchymal stem cells into insulin-secreting cells. In Vitro Cell Dev Biol Anim 2011;47(1):54-63. ##Kadam S, Muthyala S, Nair P, Bhonde R. Human placenta-derived mesenchymal stem cells and islet-like cell clusters generated from these cells as a novel source for stem cell therapy in diabetes. Rev Diabet Stud 2010;7(2):168-182.##Sun Y, Chen L, Hou XG, Hou WK, Dong JJ, Sun L, et al. Differentiation of bone marrow-derived mesenchymal stem cells from diabetic patients into insulin-producing cells in vitro. Chin Med J (Engl) 2007;120(9):771-776. ##Xie QP, Huang H, Xu B, Dong X, Gao SL, Zhang B, et al. Human bone marrow mesenchymal stem cells differentiate into insulin-producing cells upon microenvironmental manipulation in vitro. Differentiation 2009;77(5):483-491.##Phadnis SM, Joglekar MV, Dalvi MP, Muthyala S, Nair PD, Ghaskadbi SM, et al. Human bone marrow-derived mesenchymal cells differentiate and mature into endocrine pancreatic lineage in vivo Cytotherapy 2011;13(3):279-293. ##Li M, Abraham NG, Vanella L, Zhang Y, Inaba M, Hosaka N, et al. Successful modulation of type 2 diabetes in db mice with intra-bone marrow–bone marrow transplantation plus concurrent thymic transplantation. J Autoimmun 2010;35(4):414-423.##Trovato L, De Fazio R, Annunziata M, Sdei S, Favaro E, Ponti R, et al. Pluripotent stem cells isolated from human amniotic fluid and differentiation into pancreatic beta-cells. J Endocrinol Invest 2009;32(11):873-876.##Chao KC, Chao KF, Fu YS, Liu SH. Islet-like clusters derived from mesenchymal stem cells in Wharton's Jelly of the human umbilical cord for transplantation to control type 1 diabetes. PLoS One 2008;3(1):e1451. ##Wu LF, Wang NN, Liu YS, Wei X. Differentiation of Wharton's jelly primitive stromal cells into insulin-producing cells in comparison with bone marrow mesenchymal stem cells Tissue Eng Part A 2009;15(10):2865-2873.##Kadam SS, Sudhakar M, Nair PD, Bhonde RR. Reversal of experimental diabetes in mice by transplantation of neo-islets generated from human amnion derived mesenchymal stromal cells using immuno-isolatory macrocapsules. Cytotherapy 2010;12(8):982-991.##Chandra V, G S, Phadnis S, Nair PD, Bhonde RR. Generation of pancreatic hormone-expressing islet-like cell aggregates from murine adipose tissue-derived stem cells. Stem Cells 2009;27(8):1941-1953. ##Zhang YH, Wang HF, Liu W, Wei B, Bing LJ, Gao YM. Insulin-producing cells derived from rat bone marrow and their autologous transplantation in the duodenal wall for treating diabetes. Anat Rec (Hoboken) 2009;292(5):728-735.##Wong RS. Mesenchymal stem cells: angels or demons? J Biomed Biotechnol 2011;2011:459510.##Rackham CL, Dhadda PK, Chagastelles PC, Simpson SJ, Dattani AA, Bowe JE, et al. Pre-culturing islets with mesenchymal stromal cells using a direct contact configuration is beneficial for transplantation outcome in diabetic mice. Cytotherapy 2013;15(4):449-459.##Ito T, Itakura S, Todorov I, Rawson J, Asari S, Shintaku J, et al. Mesenchymal stem cell and islet co-transplantation promotes graft revascularization and function. Transplantation 2010;89(12):1438-1445. ##Lin P, Chen L, Yang N, Sun Y, Xu YX. Evaluation of stem cell differentiation in diabetic rats transplanted with bone marrow mesenchymal stem cells. Transplant Proc 2009;41(5):1891-1893. ##Kodama M, Takeshita F, Kanegasaki S, Ochiya T, Quinn G. Pancreatic endocrine and exocrine cell ontogeny from renal capsule transplanted embryonic stem cells in streptozocin-injured mice. J Histochem Cytochem 2008;56(1):33-44. ##Kodama M, Tsukamoto K, Yoshida K, Aoki K, Kanegasaki S, Quinn G. Embryonic stem cell transplantation correlates with endogenous neurogenin 3 expression and pancreas regeneration in streptozotocine-injured mice. J Histochem Cytochem 2009;57(12):1149-1158.##Ratajczak MZ, Zuba-Surma EK, Wojakowski W, Ratajczak J, Kucia M. Bone marrow-home of versatile stem cells. Transfus Med Hemother 2008;35(3):248-259.##Huang Y, Kucia M, Hussain LR, Wen Y, Xu H, Yan J, et al. Bone marrow transplantation temporarily improves pancreatic function in streptozotocin-induced diabetes: potential involvement of very small embryonic-like cells. Transplantation 2010;89(6):677-685.##Chu K, Kim M, Jeong SW, Kim SU, Yoon BW. 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ADSCs on PLLA/PCL Hybrid Nanoscaffold and Gelatin Modification: Cytocompatibility and Mechanical Properties 2 2 Background: Development of tissue engineering and regenerative medicine has led to designing scaffolds and their modification to provide a better microenvironment which mimics the natural niche of the cells. Gelatin surface modification was applied to improve scaffold flexibility and cytocompatibility. Methods: PLLA/PCL aligned fibrous scaffold was fabricated using electrospinning method. ADSCs were seeded after O2 plasma treatment and gelatin coating of the scaffolds. The morphological and mechanical properties of blends were assessed by Scanning Electron Microscopy (SEM), tensile test and ATR-FTIR. The cells proliferation was evaluated by MTT assay.  Results: Based on the results, it is supposed that gelatin coating is a brilliant method of surface modification which significantly increases the mechanical properties of scaffold without any changes on the construction or on the direction of nanofibers which conducts cell’s elongation. MTT analysis exhibited that ADSCs attachment, viability and proliferation significantly (p<0.05) increased after gelatin treatment. Conclusion: Gelatin surface modification is a highly beneficial method to improve cytocompatibility, flexibility and mechanical features of the scaffolds which doesn’t affect the nanofibers construction. Proliferation of Adipose Derived Stem Cells (ADSCs) as a remarkable source of stem cells was investigated for the first time on PLLA/PCL hybrid scaffold. 32 38 Maedeh Mashhadikhan Department of Biology, Faculty of Sciences, Science and Research Branch, Islamic Azad University Department of Biology, Faculty of Sciences, Science and Research Branch, Islamic Azad University Iran Masoud Soleimani Department of Hematology and Blood Banking, Faculty of Medical Sciences, Tarbiat Modares University Department of Hematology and Blood Banking, Faculty of Medical Sciences, Tarbiat Modares University Iran Kazem Parivar Department of Biology, Faculty of Sciences, Science and Research Branch, Islamic Azad University Department of Biology, Faculty of Sciences, Science and Research Branch, Islamic Azad University Iran Marjan Yaghmaei Department of Biology, Faculty of Sciences, Science and Research Branch, Islamic Azad University Department of Biology, Faculty of Sciences, Science and Research Branch, Islamic Azad University Iran GelatinTissue engineeringTissue scaffold 198.pdf Xu C, Lu W, Bian S, Liang J, Fan Y, Zhang X. Porous collagen scaffold reinforced with surfaced activated PLLA nanoparticles. ScientificWorldJournal 2012;2012:695137.##Cardoso GB, Perea GN, D’Avila MA, Dias CG, Zavaglia CA, Arruda AC. Initial study of electrospinning PCL/ PLLA blends. Adv Mater Phys Chem 2011;1(3):94-98.##Zhao P, Jiang H, Pan H, Zhu K, Chen W. Biodegradable fibrous scaffolds composed of gelatin coated poly(epsilon-caprolactone) prepared by coaxial electrospinning. J Biomed Mater Res A 2007;83(2):372-382.##Shabani I, Haddadi-Asl V, Seyedjafari E, Soleimani M. Cellular infiltration on nanofibrous scaffolds using a modified electrospinning technique. Biochem Biophys Res Commun 2012;423(1):50-54.##Shabani I, Haddadi-Asl V, Seyedjafari E, Babaeijandaghi F, Soleimani M. Improved infiltration of stem cells on electrospun nanofibers. Biochem Biophys Res Commun 2009;382(1):129-133.##He L, Liu B, Xipeng G, Xie G, Liao S, Quan D, et al. Microstructure and properties of nano-fibrous PCL-b-PLLA scaffolds for cartilage tissue engineering. Eur Cell Mater 2009;18:63-74.##Choi NS, Kim CH, Cho KY, Park JK. Morphology and hydrolysis of PCL/PLLA blends compatibilized with P (LLA‐co‐CL) or P (LLA‐b‐CL). J Appl Polym Sci 2002;86(8):1892-1898.##Paul MA, Alexandre M, Degée P, Henrist C, Rulmont A, Dubois P. New nanocomposite materials based on plasticized poly (L-lactide) and organo-modified montmorillonites: thermal and morphological study. Polymer 2003;44(2):443-450.##Todo M, Harada A, Tsuji H. Fracture characterizarion of biodegradable PLLA polymer blends. 16th International conference on composite materials. Kyoto, 8-13 July 2007. p. 1-6.##Klump D, Horch RE, Kneser U, Beier JP. Tissue engineering of skeletal muscle. J Cell Mol Med 2010;14(11):2622-2629.##Chen JP, Su CH. Surface modification of electrospun PLLA nanofibers by plasma treatment and cationized gelatin immobilization for cartilage tissue engineering. Acta Biomater 2011;7(1):234-243.##Jahani H, Kaviani S, Hassanpour-Ezatti M, Soleimani M, Kaviani Z, Zonoubi Z. The effect of aligned and random electrospun fibrous scaffolds on rat mesenchymal stem cell proliferation. Cell J 2012;14(1):31-38.##Guo C, Cai N, Dong Y. Duplex surface modification of porous poly (lactic acid) scaffold. Mater Lett 2013;94:11-14.##Kim MS, Jun I, Shin YM, Jang W, Kim SI, Shin H. The development of genipin-crosslinked poly(caprolactone) (PCL)/gelatin nanofibers for tissue engineering applications. Macromol Biosci 2010;10(1):91-100.##Htay MAS. Water vapour transmission and degradation properties of biaxially stretched PCL films and cell-permeable membranes [master's thesis]. [Singapore]: National University of Singapore 2004. 130 p.##Jin W, Liu L. Micro construction of poly(epsilon-caprolactone)/poly(L-lactic acid) blend film by solution casting under microwave irradiation. Macromol Biosci 2004;4(7):656-664.##Can E, Udenir G, Kanneci AI, Kose G, Bucak S. Investigation of PLLA/PCL blends and paclitaxel release profiles. AAPS PharmSciTech 2011;12(4):1442-1453.##Chamy R, Rosenkranz F. Biodegradation of hazardous and special products [Internet]. Croatia: InTech; 2013 Jun 14. Kodama Y. Degradability: enzymatic and in simulated compost soil of PLLA: PCL blend and on their composite with coconut fiber; [64 pages]. Availiable from: http://www.intechopen.com/books/biodegradation-of-hazardous-and-special-products/degradability-enzymatic-and-in-simulated-compost-soil-of-plla-pcl-blend-and-on-their-composite-with-.##Patricio T, Gloria A, Bartolo P. Mechanical and biological behaviour of PCL and PCL/PLA scaffolds for tissue engineering applications. Chem Eng Trans 2013;32:1645-1650.##Yao M, Deng H, Mai F, Wang K, Zhang Q, Chen F, et al. Modification of poly (lactic acid)/poly (propylene carbonate) blends through melt compounding with maleic anhydride. Express Polym Lett 2011;5:937.##Shang S, Yang F, Cheng X, Walboomers XF, Jansen JA. The effect of electrospun fibre alignment on the behaviour of rat periodontal ligament cells. Eur Cell Mater 2010;19:180-192.##Tseng LF, Mather PT, Henderson JH. Shape-memory-actuated change in scaffold fiber alignment directs stem cell morphology. Acta Biomater 2013;9(11):8790-8801. ##Chew SY, Mi R, Hoke A, Leong KW. The effect of the alignment of electrospun fibrous scaffolds on Schwann cell maturation. Biomaterials 2008;29(6):653-661.##Seyedjafari E, Soleimani M, Ghaemi N, Shabani I. Nanohydroxyapatite-coated electrospun poly (l-lactide) nanofibers enhance osteogenic differentiation of stem cells and induce ectopic bone formation. Biomacromolecules 2010;11(11):3118-3125.##Lin Y, Wang L, Zhang P, Wang X, Chen X, Jing X, et al. Surface modification of poly (L-lactic acid) to improve its cytocompatibility via assembly of polyelectrolytes and gelatin. Acta Biomate 2006;2(2):155-64.##Oraby MA, Waley AI, El-Dewany AI, Saad EA, Abd El-Hady BM. Electrospinning of gelatin functionalized with silver nanoparticles for nanofiber fabrication. Model Numer Simul Mater Sci 2013;3(4):95-105.##Desimone D, Li W, Roether JA, Schubert DW, Crovace MC, Rodrigues ACM, et al. Biosilicate®–gelatine bone scaffolds by the foam replica technique: development and characterization. Sci Technol Adv Mater 2013;14(4):045008.##Pignatello R. Biomaterials applications for nanomedicine [Internet]. Croatia: InTech; 2011 16 Nov. Gorgieva S, Kokol V. Collagen-vs. gelatine-based biomaterials and their biocompatibility: Review and perspectives; [37 pages]. Availiable from: http://www.intechopen.com/books/biomaterials-applications-for-nanomedicine/collagen-vs-gelatine-based-biomaterials-and-their-biocompatibility-review-and-perspectives.##Houa X, Zhangb BL, Shea F, Cuia YL, Shib LY, Yaoa KD. Surface of gelatin modified poly (L-lactic acid) film. Chinese J Polym Sci 2003;21(3):277-283.##Cui YL, Hou X, Qi AD, Wang XH, Wang H, Cai KY, et al. 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Cloning and Expression of CD19, a Human B-Cell Marker in NIH-3T3 Cell Line 2 2 Background: CD19 is a pan B cell marker that is recognized as an attractive target for antibody-based therapy of B-cell disorders including autoimmune disease and hematological malignancies. The object of this study was to stably express the human CD19 antigen in the murine NIH-3T3 cell line aimed to be used as an immunogen in our future study. Methods: Total RNA was extracted from Raji cells in which high expression of CD19 was confirmed by flow cytometry. Synthesized cDNA was used for CD19 gene amplification by conventional PCR method using Pfu DNA polymerase. PCR product was ligated to pGEM-T Easy vector and ligation mixture was transformed to DH5α competent bacteria. After blue/white selection, one positive white colony was subjected to plasmid extraction and direct sequencing. Then, CD19 cDNA was sub-cloned into pCMV6-Neo expression vector by double digestion using KpnI and HindIII enzymes. NIH-3T3 mouse fibroblast cell line was subsequently transfected by the construct using Jet-PEI transfection reagent. After 48 hours, surface expression of CD19 was confirmed by flow cytometry and stably transfected cells were selected by G418 antibiotic. Results: Amplification of CD19 cDNA gave rise to 1701 bp amplicon confirmed by alignment to reference sequence in NCBI database. Flow cytometric analysis showed successful transient and stable expression of CD19 on NIH-3T3 cells (29 and 93%, respectively). Conclusion: Stable cell surface expression of human CD19 antigen in a murine NIH-3T3 cell line may develop a proper immunogene which raises specific anti-CD19 antibody production in the mice immunized sera. 39 44 Hajar Abbasi-Kenarsari Immunology Research Center, Tabriz University of Medical Sciences, Department of Immunology, School of Medicine, Tabriz University of Medical SciencesStudents Research Committee, Tabriz University of Medical Sciences Immunology Research Center, Tabriz University of Medical Sciences, Department of Immunology, School of Medicine, Tabriz University of Medical SciencesStudents Research Committee, Tabriz University of Medical Sciences IranIranIran Farzaneh Shafaghat Department of Immunology, International Branch of Aras, Tabriz University of Medical Sciences Department of Immunology, International Branch of Aras, Tabriz University of Medical Sciences Iran Behzad Baradaran Immunology Research Center, Tabriz University of Medical SciencesDepartment of Immunology, School of Medicine, Tabriz University of Medical Sciences Immunology Research Center, Tabriz University of Medical SciencesDepartment of Immunology, School of Medicine, Tabriz University of Medical Sciences IranIran Ali Akbar Movassaghpour Hematology and Oncology Research Center, Tabriz University of Medical Sciences Hematology and Oncology Research Center, Tabriz University of Medical Sciences Iran Dariush Shanehbandi Immunology Research Center, Tabriz University of Medical Sciences Immunology Research Center, Tabriz University of Medical Sciences Iran Tohid Kazemi Immunology Research Center, Tabriz University of Medical SciencesDepartment of Immunology, School of Medicine, Tabriz University of Medical Sciences Immunology Research Center, Tabriz University of Medical SciencesDepartment of Immunology, School of Medicine, Tabriz University of Medical Sciences IranIran B cellCD19CloningGene expression 199.pdf Blanc V, Bousseau A, Caron A, Carrez C, Lutz RJ, Lambert JM. 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Use of Raman Spectroscopy to Decrease Time for Identifying the Species of Candida Growth in Cultures 2 2 Background: The objective of this study is to establish Raman signatures from pure cultures of different Candida species using Raman Spectroscopy (RS) and use these signatures for rapid identification of unknown Candida species. Methods: Pure cultures of five Candida species were evaluated using RS to build a limited signature library. ‘Raman Processing’ (RP) software was used for Principal Component Analysis (PCA) and Differential Functional Analysis (DFA). Results: Eleven principal components described at least 95% variance in the spectra. Raman signatures from these known Candida species were able to identify the species of unknown Candida cultures with 100% accuracy. Conclusion: Raman spectroscopy can improve early identification of Candida species and may facilitate early optimal antifungal therapy.   45 48 Nitin S. Chouthai Division of Neonatal-Perinatal Medicine, Wayne State University Division of Neonatal-Perinatal Medicine, Wayne State University United States of America Anuj A. Shah Division of Neonatal-Perinatal Medicine, Wayne State University Division of Neonatal-Perinatal Medicine, Wayne State University United States of America Hossein Salimnia Division of Pathology, Wayne State University Division of Pathology, Wayne State University United States of America Olena Palyvoda Lumigen Instrument Center, Wayne State University Lumigen Instrument Center, Wayne State University United States of America Suneetha Devpura Department of Radiation Oncology, Henry Ford Health System Department of Radiation Oncology, Henry Ford Health System United States of America Michael Klein Pediatric Surgery, Wayne State University Pediatric Surgery, Wayne State University United States of America Basim Asmar Division of Pediatric Infectious Diseases, Wayne State University Division of Pediatric Infectious Diseases, Wayne State University United States of America Candida speciesRamanSpectrum analysis 200.pdf Stoll BJ, Hansen N, Fanaroff AA, Wright LL, Carlo WA, Ehrenkranz RA, et al. 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Indian J Med Microbiol 2012;30(3):264-269. ##Trovato L, Betta P, Romeo MG, Oliveri S. Detection of fungal DNA in lysis–centrifugation blood culture for the diagnosis of invasive candidiasis in neonatal patients. Clin Microbiol Infect 2012;18(3):E63-E5.##Morris AJ, Byrne TC, Madden JF, Reller LB. Duration of incubation of fungal cultures. J Clin Microbiol 1996;34(6):1583-1585.##