Editorial 2 2 The seventh National Biotechnology Congress in Iran was held in Tehran from 12-14 September 2011. Since many of our readers most likely did not find the opportunity to attend this conference, I take this opportunity to provide a summary of activities in this three day congress. In the seventh National Biotech-nology Congress in Iran, dozens of talks and several hundred posters were presented. The topics of biotech-nology-related subjects presented in this congress were wide spread and included: Transgenic animals, plant biotechnology, systems biology, next generation DNA sequencing, biotech market, biotech manage-ment, environment, pharmaceuticals, food industry, medical biotechnology, marine biotechnology, bio-informatics and ethics. This congress was attended by hundreds of students, researchers, government officials and industry leaders from all corners of Iran. This congress was sponsored and/or supported by more than 25 national research centers, private institutes and government organizations including the presidential office. Holding regular National biotechnology meetings in the field of biotechnology in Iran is hoped to deepen the discussions of biotech-related issues in both academic and non-academic centers and encourage inter-actions between the interested parties. An interesting and novel aspect of this congress was an attempt to initiate a dialogue between the biotech leaders and religious authorities. The reality is that application of biotechnologies in different disciplines including genetic manipulation of plant and animal cells, food, … has caused anxieties and social concerns in recent years. Therefore, any attempt to encourage dialogues between the biotech and religious leaders should be helpful in presenting biotechnology as a helpful technology to solve societal problems. Today biotechnology is one of the fastest growing technologies worldwide as was predicted a couple of decades ago. The market value of biotech-related technologies is estimated to be over $ 1000 billion worldwide and is dominated by the developed countries mainly due to the massive investments they did in biotech industry a couple of decades earlier. The question is whether the developing countries can develop the biotech industry segments of their economies soon enough to capture a portion of this huge market worldwide. Hopefully holding the seventh National biotechnology meeting in Iran would focus the attention of the scientific, economic and political leaders on this important issue. 156 156 Ali M. Ardekani Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Editorial 164.pdf ####

Stress-Induced Proteomic Changes in the Hippocampus of Pregnant Wistar Rats 2 2 Stress is a threatening factor that all living organisms encounter throughout life. Depending on the type of stress, there are several mechanisms for keeping body homeostasis to minimize stress effects. Brain is an organ which shows high sensitivity to stress conditions. Although many studies have shown induced-stress effects on rat embryos, little is known about the mechanisms involved in coping with stress by female rats during pregnancy. In the present study, restraint stress method was applied because this technique has been widely used in animal models to induce both psychological and physical stress. Restraint stress was applied in regular sessions (1 and 3 hrs) in two groups of 6 pregnant Wistar rats and similar number of animals was used as control group receiving no stress. ACTH and corticosterone levels in plasma samples were shown to increase in response to stress treatments. On the last day of pregnancy, rat hippocampus from the brain of each animal in all three groups was removed and analyzed using 2 Dimensional Gel Electrophoresis (2DE) technique. Using Image Master Software, approximately 2000 proteins were detected in the 2D gels analyzed, among which 34 proteins exhibited differential expression. These results indicate that the proteome patterns from the hippocampus of pregnant rats subjected to 1 and 3 hr of stress differs significantly from the control (unstressed) group. Future mass spectrometry identification of the 34 protein spots discovered in this study should allow a more precise understanding of molecules and cellular pathways involved in stress-induced responses during pregnancy. 157 166 Ali M. Ardekani Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Nader Maghsudi Neuroscience Research Center, Shahid Beheshti University of Medical Sciences Neuroscience Research Center, Shahid Beheshti University of Medical Sciences Iran Anna Meyfour Reproductive Biotechnology Research Center, Avicenna Research Institute Reproductive Biotechnology Research Center, Avicenna Research Institute Iran Rasool Ghasemi Neuroscience Research Center, Shahid Beheshti University of Medical Sciences Neuroscience Research Center, Shahid Beheshti University of Medical Sciences Iran Niknam Lakpour Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Elahe Nooshinfar Department of Physiology, Faculty of Paramedical Sciences, Shahid Beheshti University of Medical Sciences Department of Physiology, Faculty of Paramedical Sciences, Shahid Beheshti University of Medical Sciences Iran Zahra Ghaempanah Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran 2 Dimensional gel electrophoresisBrainHippocampusPregnancyProteomicsStress 70.pdf Latendresse G. 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Neuropsychologia 2007;45(12):2699-2711.##Sapolsky RM. The possibility of neurotoxicity in the hippocampus in major depression: a primer on neuron death. Biol Psychiatry 2000;48(8):755-765.##Grossman R, Buchsbaum MS, Yehuda R. Neuroimaging studies in post-traumatic stress disorder. Psychiatr Clin North Am 2002;25(2):317-340.##McKinnon MC, Yucel K, Nazarov A, MacQueen GM. A meta-analysis examining clinical predictors of hippocampal volume in patients with major depressive disorder. J Psychiatry Neurosci 2009;34(1):41-54.##Brummelte S, Galea LA. Chronic high corticosterone reduces neurogenesis in the dentate gyrus of adult male and female rats. Neuroscience 2010;168:680-690.##Dalla C, Antoniou K, Drossopoulou G, Xagoraris M, Kokras N, Sfikakis A, et al. Chronic mild stress impact: are females more vulnerable? Neuroscience 2005;135(3):703-714.##Kornstein SG. Gender differences in depression: implications for treatment. 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Comparison of Epothilone and Taxol Binding in Yeast Tubulin using Molecular Modeling 2 2 Microtubules are unique cytoskeletal structures that have structural subunits of αβ tubulin. Taxol is a typical microtubule stabilizing drug. The epothilones are other natural products with similar mechanism of action totaxol. Despite the highly conserved nature of β-tubulin, some organism like Saccharomyces cerevesia is resistance to taxol, but sensitive to epothilones. In order to find differences in sensitivity of yeast tubulin to these molecules, we investigated binding mode of the taxol and epothilone A to yeast tubulin using molecular modeling. The multiple sequence alignment of β-tubulin of different species was performed using ClustalW2. Protein structure of yeast β-tubulin was constructed with Swiss Model 8.05 by using 1TVK. Modeled tubulin was superimposed with PyMol on1JFF for comparison of three-dimensional structure of two proteins. Our results showed that one of the most interesting differences in binding mode of these molecules is residue 227. The His227 in bovine makes a hydrogen bond by means of its δ-nitrogen with epothilone A and by means of its ε-nitrogen with taxol. The Asn227 of yeast can play role of the δ-nitrogen of imidazole ring of H227, but not of ε-nitrogen of it. So yeast tubulin in contrast to taxol can interact with epothilone A. Due to conservation of essential residues for binding (T274, R282 and Q292), epothilone A in comparison with taxol can tolerate the interchange in the binding pocket (R276I). Our findings may be of a great aid in the rational design of anti-tumor agents that bind to the taxol binding region of tubulin. 167 176 Vajihe Akbari Department of Pharmaceutical Biotechnology, Faculty of Pharmacy, Isfahan University of Medical Science Department of Pharmaceutical Biotechnology, Faculty of Pharmacy, Isfahan University of Medical Science Iran Sharareh Moghim Department of Bacteriology and Virology, Faculty of Medicine, Isfahan University of medical sciences Department of Bacteriology and Virology, Faculty of Medicine, Isfahan University of medical sciences Iran Mohammad Reza Mofid Department of Biochemistry, School of Pharmacy and Isfahan Pharmaceutical Sciences Research Center, Isfahan University of Medical Sciences Department of Biochemistry, School of Pharmacy and Isfahan Pharmaceutical Sciences Research Center, Isfahan University of Medical Sciences Iran Epothilone AMicrotubulesSaccharomyces cerevesiaTaxolβ-tubulin 73.pdf Hyams JS, Lloyd CW. Microtubules. In: Harford JB (eds). Modern Cell Biology. Vol. 13. New York:Wiley-Liss;1994. ##He L, Orr GA, Horwitz SB. Novel molecules that interact with microtubules and have functional activity similar to Taxol. Drug Discov Today 2001;6(22):1153-1164.##Bollag, DM, McQueney PA, Zhu J, Hensens O, Koupal L, Liesch J, et al. Epothilones, a new class of microtubule-stabilizing agents with a taxol-like mechanism of action. Cancer Res 1995;55(11):2325-2333.##Kowalski R J, Giannakakou P, Hamel E. Activities of the microtubule-stabilizing agents epothilones A and B with purified tubulin and in cells resistant to paclitaxel (Taxol®). J Biol Chem 1997;272(4): 2534-2541.##Gull K. Protist tubulins: new arrivals, evolutionary relationships and insights into cytoskeletal function. Curr Opin Microbiol 2001;4(4):427-432.##McKean PG, Vaughan S, Gull K. The extended tubulin superfamily. J Cell Sci 2001;114(Pt 15):2723-2733.##Gupta ML, Bode CJ, Georg GI, Himes RH. 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Bryostatin-1, Fenretinide and 1α,25 (OH)2D3 Induce Growth Inhibition, Apoptosis and Differentiation in T and B Cell-Derived Acute Lymphoblastic Leukemia Cell Lines (CCRF-CEM and Nalm-6) 2 2 In many acute leukemias, normal differentiation does not occur. However, in many cell lines derived from hematologic malignancies, differentiation or apoptosis can be induced by variety of agents. Despite advances in the treatment of Acute Lymphoblastic Leukemia (ALL), in most patients long-term survival rates remain unsatisfactory, especially in T-cell derived ALL. Thus we studied the anti-cancer effects of fenretinide, 1α,25(OH)2D3, and bryostatin-1 in CCRF-CEM (T-cell derived) and Nalm-6 (B-cell derived) ALL cell lines. Using MTT assays, both cell lines were shown to exhibit increased inhibition of proliferation at micro (fenretinide) and nanomolar (1α,25(OH)2D3, bryostatin-1) concentrations. These anti-cancer agents were shown to induce apoptosis and activate caspase-3 pathway in both ALL cell lines. Furthermore, for the first time we are reporting consistent anti-proliferative and apoptotic effects of Bryostatin-1 in ALL T-cell derived cell line with the lowest ED50 (ranging 4.6 nM - 7.4 nM). To evaluate the differentiation induction by fenretinide, 1α,25(OH)2D3, and bryostatin-1 in ALL cell lines, we assayed for the expressions of CD19, CD38 markers on Nalm-6 and CD7 marker on CCRF-CEM cell line. The flow cytometric analysis showed a significant increase in expression of CD markers in response to anticancer drug treatments. To assay the effects of anti-cancer drugs on cell cycle distribution, cell cycle analysis using flow cytometry was employed. These anti-cancer drugs appear to affect the CCRF-CEM and Nalm-6 cell cycles differently (G0/G1 and G2/M, respectively). Overall results demonstrate that the anticancer agents used in this study are strong inhibitors of ALL cell proliferation and inducers of apoptosis and differentiation in vitro. These findings may be quite helpful if these drugs are to be used for differentiation therapy of ALL patients in clinics in the future. Further studies are warranted to establish the in vivo effect of these drugs particularly in patients with T-cell derived ALL. 177 194 Ali M. Ardekani Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Shahrzad Soleymani Fard Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Mahmood Jeddi-Tehrani Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Iran Ramin Ghahremanzade Nanotechnology Research Center, Avicenna Research Institute, ACECR Nanotechnology Research Center, Avicenna Research Institute, ACECR Iran Acute lymphoblastic leukemiaApoptosisCell differentiationFlow cytometry 71.pdf Estey E. Treatment of relapsed and refractory acute myelogenous leukemia. Leukemia 2000;14(3):476-479.##Cheok MH, Evans WE. Acute lymphoblastic leukaemia: a model for the pharmacogenomics of cancer therapy. 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Altered nuclear receptor corepressor expression attenuates vitamin D receptor signaling in breast cancer cells. Clin Cancer Res 2006;12(7 Pt 1):2004-2013.##Yan W, Chen W, Liu Z, Huang L. Bryostatin-I: A dendritic cell stimulator for chemokines induction and a promising adjuvant for a peptide based cancer vaccine. Cytokine 2010;52(3):238-244.##Clamp A, Jayson GC. The clinical development of the bryostatins. Anticancer Drugs. 2002;13(7):673-683.##Wang S, Guo C-Y, Castillo A, Dent P, Grant S. Effect of bryostatin 1 on taxol-induced apoptosis and cytotoxicity in human leukemia cells (U937). Biochem Pharmacol 1998;56(5):635-644.##Asiedu C, Biggs J, Lilly M, Kraft AS. Inhibition of leukemic cell growth by the protein kinase C activator bryostatin 1 correlates with the dephosphorylation of cyclin-dependent kinase 2. Cancer Res 1995;55(17):3716-3720.##Roberts JD, Smith MR, Feldman EJ, Cragg L, Millenson MM, Roboz GJ. Phase I study of bryostatin 1 and fludarabine in patients with chronic lymphocytic leukemia and indolent (non-Hodgkin's) lymphoma. Clin Cancer Res 2006;12 (19):5809-5816.##Faderl S, Lotan R, Kantarjian HP, Harris D, Van Q, Estrov Z. N-(4 Hydroxylphenyl)retinamide (fenretinide, 4-HPR), a retinoid compound with antileukemic and proapoptotic activity in acute lymphoblastic leukemia (ALL). Leuk Res 2003;27(3):259-266.##Matsui W, Smith BD, Vala M, Beal N, Huff CA, Diehl LF et al. Requirement for myeloid growth factors in the differentiation of acute promyelocytic leukaemia. Br J Haematol 2005;128(6):853-862.##Matsui WH, Gladstone DE, Vala MS, Barber JP, Brodsky RA, Smith BD et al. The role of growth factors in the activity of pharmacological differentiation agents. Cell Growth Differ 2002;13(6):275-283.##Das A, Banik NL, Ray SK. Retinoids induced astrocytic differentiation with down regulation of telomerase activity and enhanced sensitivity to taxol for apoptosis in human glioblastoma T98G and U87MG cells. J Neurooncol 2008;87(1):9-22.##Wagner KD, Wagner N, Sukhatme VP, Scholz H. Activation of vitamin D receptor by the Wilms’ tumor gene product mediates apoptosis of renal cells. J Am Soc Nephrol 2001;12(6):1188-1196.##Soleymani S, Jeddi-Tehrani M, Akhondi MM, Hashemi M, Ardekani A. Flow cytometric analysis of 4-HPR-induced apoptosis and cell cycle arrest in acute myelocytic leukemia cell line (NB-4). Avicenna J Med Biotech 2010;2(1):53-61. ##Wagner N, Wagner KD, Schley G, Badiali L, Theres H, Scholz H. 1,25-dihydroxyvitamin D-induced apoptosis of retinoblastoma cells is associated with reciprocal changes of Bcl-2 and bax. Exp Eye Res 2003;77(1):1-9.##Bastie JN, Balitrand N, Guillemot I, Chomienne C, Delva L. 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Nat Med 2005;11(6):630-637.##Hotﬁlder M, Rottgers S, Rosemann A, Schrauder A, Schrappe M, Pieters R, et al. Leukemic stem cells in childhood high-risk ALL/ t(9;22) and t(4;11) are present in primitive lymphoid-restricted CD34+CD19 cells. Cancer Res. 2005;65(4):1442-1449.##Sato S, Miller AS, Howard MC, Tedder TF. Regulation of B lymphocyte development and activation by the CD19/CD21/ CD81/Leu 13 complex requires the cytoplasmic domain of CD19. J Immunol 1997;159(7):3278-3287.##Fujimoto M, Poe JC, Hasegawa M, Tedder TF. CD19 regulates intrinsic B lymphocyte signal transduction and activation through a novel mechanism of processive ampliﬁcation. Immunol Res 2000;22(2-3):281-298.##Donis-Hernandez FR, Parkhouse RM, Santos-Argumedo L. Ontogeny, distribution and function of CD38-expressing B lymphocytes in mice. Eur J Immunol 2001;31(4):1261-1267.##Springer LN, Stewart BW. N-(4-Hydroxyphenyl) retinamide-induced death in human lymphoblastoid cells: 50 kb DNA breakage as a means of distinguishing apoptosis from necrosis. Cancer Lett 1998;128(2):189-196.##Cao J, Xu D, Wang D, Wu R, Zhang L, Zhu H, et al. ROS-driven Akt dephosphorylation at Ser-473 is involved in 4-HPR-mediated apoptosis in NB4 cells. Free Radic Biol Med 2009;47(5):536-547.##Clark CS, Konyer JE, Meckling KA. 1α,25-dihydroxyvitamin D3 and bryostatin-1 synergize to induce monocytic differentiation of NB4 acute promyelocy tic leukemia cells by modulating cell cycle progression. Exp Cell Res 2004;294(1):301-311.##Steube KG, Grunicke D, Ouentmeier H, Drexler HG. Different effects of the two protein kinase C activator bryostatin-1 and TPA on growth and differentiation of human monocytic leukemia cell lines. Leuk Res 1993;17(10):897-901.##Chelliah J, Freemerman AJ, Wu-Pang S, Jawis WD, Grant S. Potentiation of ara-C-induced apoptosis by the protein kinase C activator bryostatin 1 in human leukemia cells (HL-60) involves a process dependent upon c-Myc. Biochem Pharmacol 1997;54(5):563-573.##Jones RJ, Sharkis SJ, Miller CB, Rowinsky EK, Burke P, May WS. Bryostatin 1, a unique biologic response modifier: anti-leukemic activity in vitro. Blood 1990;75(6):1319-1323. ##Liu M, Lee MH, Cohen M, Bommakanti M, Freedman LP. Transcriptional activation of the Cdk inhibitor p21 by vitamin D3 leads to the induced differentiation of the myelomonocytic cell line U937. Genes Dev 1996;10(2):142-153.##O'Donnell PH, Guo WX, Reynolds CP, Maurer BJ. N-(4-hydroxyphenyl) retinamide increases ceramide and is cytotoxic to acute lymphoblastic leukemia cell lines, but not to non-malignant lymphocytes. Leukemia 2002;16(5):902-910.##Zhang X, Zhang R, Zhao H, Cai H, Gush K, Kerr R, et al. Preclinical pharmacology of the natural product anticancer agent bryostatin 1, an activator of protein kinase C. Cancer Res 1996;56(4):802-808.##Kortmansky J, Schwartz G. Bryostatin-1: A novel PKC inhibitor in clinical development. Cancer Invest 2003;21(6):924-936.##Jarvis WD, Povirk LF, Turner AJ, Traylor RS, Gewirtz DA, Pettit GR, et al. Effects of bryostatin 1 and other pharmacological activators of protein kinase C on 1-(beta-D- arabinofuranosyl)cytosine-induced apoptosis in HL-60 human promyelocytic leukemia cells. Biochem Pharmacol 1994;47(5):839-852.##Grant S, Turner AJ, Freemerman AJ, Wang Z, Kramer L, Jarvis WD. Modulation of protein kinase C activity and calcium-sensitive isoform expression in human myeloid leukemia cells by bryostatin 1: relationship to differentiation and ara-C-induced apoptosis. Exp Cell Res 1996;228(1): 65-75.##Vrana JA, Saunders AM, Chellappan SP, Grant S. Divergent effects of bryostatin 1 and phorbol myristate acetate on cell cycle arrest and maturation in human myelomonocytic leukemia cells (U937). Differentiation 1998;63(1):33-42.##Wall NR, Mohammad RM, Nabha SM, Pettit GR, Al-Katib AM. Modulation of cIAP-1 by novel antitubulin agents when combined with bryostatin 1 result in increased apoptosis in the human early pre-B acute lymphoblastic leukemia cell line Reh. Biochem Biophys Res Commun 1999;266 (1):76-80.##Leszczyniecka M, Roberts T, Dent P, Grant S, Fisher PB. Differentiation therapy of human cancer: basic science and clinical applications. Pharmacol Ther 2001;90(2-3):105-156. ##Studzinski GP, Harrison LE. Differentiation-related changes in the cell cycle traverse. Int Rev Cytol 1999;189:1-58.##Wu JM, DiPietrantonio AM, Hsieh TC. Mechanism of fenretinide (4-HPR)-induced cell death. Apoptosis 2001;6(5):377-388.##Rao AS, Freemerman AJ, Jarvis WD, Chelliah J, Bear HD, Mikkelsen R, et al. Effect of AS101 on bryostatin 1-mediated differentiation induction, cell cycle arrest, and modulation of drug-induced apoptosis in human myeloid leukemia cells. Leukemia 1996;10(7): 1150-1158.##Bastie JN, Balitrand N, Guidez F, Guillemot I, Larghero J, Calabresse C, et al. 1α,25-dihydroxyvitamin D3 transrepresses retinoic acid transcriptional activity via vitamin D receptor in myeloid cells. Mol Endocrinol 2004;18(11):2685-2699.##Janardhanan R, Butler JT, Banik NL, Ray SK. N-(4-Hydroxyphenyl) retinamide potentiated paclitaxel for cell cycle arrest and apoptosis in glioblastoma C6 and RG2 cells. Brain Res 2009;1268:142-153.##Reddy CD, Guttapalli A, Adamson PC, Vemuri MC, O’Rourke D, Sutton LN, et al. Anticancer effects of fenretinide in human medulloblastoma. Cancer Lett 2006;231(2):262-269.##Janardhanan R, Banik NL, Ray SK. N-(4-Hydroxyphenyl) retinamide induced diﬀerentiation with repression of telomerase and cell cycle to increase interferon-c sensitivity for apoptosis in human glioblastoma cells. Cancer Lett 2008;261(1):26-36.##Wang S, Wang Z, Boise LH, Dent P, Grant S. Bryostatin 1 enhances paclitaxel-induced mitochondrial dysfunction and apoptosis in human leukemia cells (U937) ectopically expressing Bcl-x. Leukemia 1999;13(10):1564-1573.##Mohammad RM, Diwakaran H, Maki A, Emara MA, Pettit GR, Redman B, et al. Bryostatin 1 induced apoptosis and augments inhibitory effects of Vincristine in human diffuse large cell lymphoma. Leuk Res 1995;19(9):667-673.##Artaza JN, Sirad F, Ferrini MG, Norris KC. 1α,25(OH)2vitamin D3 inhibits cell proliferation by promoting cell cycle arrest without inducing apoptosis and modiﬁes cell morphology of mesenchymal multipotent cells. J Steroid Biochem Mol Biol 2010;119(1-2):73-83.##Elstner E, Linker-Israeli M, Umiel T, Le J, Grillier I, Said J, et al. Combination of a potent 20-epi-vitamin D3 analogue (KH1060) with 9-cis retinoic acid irreversibly inhibits clonal growth, decreases bcl-2 expression, and induces apoptosis in HL-60 leukemic cells. Cancer Res 1996;56(15):3570-3576.##James SY, Turner A, Colston KW. 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An Enhancing Effect of Gold Nanoparticles on the Lethal Action of 2450 MHz Electromagnetic Radiation in Microwave Oven 2 2 Today, there is an increasing interest in the use of metal nanoparticles in health sciences. Amongst all nanoparticles, the gold nanoparticles have been known to kill the cancer cells under hyperthermic condition by near-infrared frequency electromagnetic waves. On the other hand, although there are different physiochemical methods for disinfection of microbial pollution, however applications of irradiated gold nanoparticles against microorganisms have not yet been investigated. In this study, gold nanoparticles were prepared using D-glucose and characterized (particle size <26 nm). In the next step, the enhancing effect of the non toxic level of gold nanoparticles (50 µg/mL) on the antimicrobial activity of 2450 MHz electromagnetic radiation generated at a microwave oven operated at low power (100 W), was investigated by time-kill course assay against Staphylococcus aureus (S. aureus) ATCC 29737. The results showed that application of gold nanoparticles can enhance the lethal effect of low power microwave in a very short exposure time (5 s). 195 200 Kamyar Mollazadeh-Moghaddam Students' Scientific Research Center, Tehran University of Medical Sciences Students' Scientific Research Center, Tehran University of Medical Sciences Iran Bardia Varasteh Moradi Department of Pharmaceutical Biotechnology and Biotechnology Research Center, Faculty of Pharmacy, Tehran University of Medical Sciences Department of Pharmaceutical Biotechnology and Biotechnology Research Center, Faculty of Pharmacy, Tehran University of Medical Sciences Iran Reza Dolatabadi-Bazaz Department of Medicinal Chemistry, Faculty of Pharmacy, Tehran University of Medical Sciences Department of Medicinal Chemistry, Faculty of Pharmacy, Tehran University of Medical Sciences Iran Mojtaba Shakibae Department of Pharmacognosy and Biotechnology, School of Pharmacy, Pharmaceutics Research Center, Kerman University of Medical Sciences Department of Pharmacognosy and Biotechnology, School of Pharmacy, Pharmaceutics Research Center, Kerman University of Medical Sciences Iran Ahmad Reza Shahverdi Department of Pharmaceutical Biotechnology and Biotechnology Research Center, Faculty of Pharmacy, Tehran University of Medical Sciences Department of Pharmaceutical Biotechnology and Biotechnology Research Center, Faculty of Pharmacy, Tehran University of Medical Sciences Iran Antimicrobial effectElectromagnetic radiationGold nanoparticlesMicrowave<i>Staphylococcus aureus</i> 75.pdf Gier LJ. Effects of ultrashort radio waves and ultraviolet light on microorganisms. Trans Kans Acad Sci 1937;40(1-3):55-57.##Garcia MM, Brooks BW, Stewart RB, Dion W, Trudel JR, Ouwerkerk T. Evaluation of gamma radiation levels for reducing pathogenic bacteria and fungi in animal sewage and laboratory effluents. Can J Vet Res 1987;51(3): 285-289.##Morgan WF, Day JP, Kaplan MI, McGhee EM, Limoli CL. Genomic instability induced by ionizing radiation. Radiat Res 1996;146(3):247-258.##Leonard A, Berteaud AJ, Bruyere A. An evaluation of the mutagenic, carcinogenic and teratogenic potential of microwaves. Mutat Res 1983;123(1):31-46.##Geveke DJ, Brunkhorst C, Fan X. Radio frequency electric fields processing of orange juice. Innovat Food Sci Emerg Tech 2007;8(4):549-554.##Wiesbrock F, Hoogenboom R, Schubert US. Microwave-assisted polymer synthesis: State-of-the-art and future prespective. Macromol Rapid Commun 2004;25:1739-1764.##Riddle MM, Smialowicz RJ Rogers RR. Microwave radiation (2450-MHz) potentiates the lethal effect of endotoxin in mice. Health Phys 1982;42(3): 335-340.##Vela G R, Wu JF. Mechanism of lethal action of 2,450-MHz radiation on microorganisms. Appl Environ Microbiol 1979;37 (3):550-553.##Ohlsson T, Bengtsson N. Microwave technology and foods. Adv Food Nutr Res 2001;43:65-140.##Bialoszewski D, Bocian E, Bukowska B, Czajkowska M, Sokol-Leszczynska B, Tyski S. Antimicrobial activity of ozonted water. Med Sci Monit 2010; 16 (9):MT71-5.##Bryce EA, Spence D, Roberts FJ. An in-use evaluation of an alcohol-based pre-surgical hand disinfectant. Infect Contr Hosp Epidemiol 2001;22(10):635-639.##Rai M, Yadav A, Gade A. Silver nanoparticles as a new generation of antimicrobials. Biotechnol Adv 2009;27(1):76-83.##Arshi N, Ahmed F, Kumar S, Anwar MS, Lu J, Koo BH , et al. Microwave assisted synthesis of gold nanoparticles and their antibacterial activity against Escherichia coli (E.coli). Curr Appl Phys 2011;11(1 Supplement):S360-S363. ##Huff TB, Tong L, Zhao Y, Hansen MN, Cheng JX, Wei A. Hyperthermic effects of gold nanorods on tumor cells. Nanomedicine 2007;2(1):125-132.##Raveendran P, Fu J, Wallen SL. A simple and “green” method for the synthesis of Au, Ag, and Au–Ag alloy nanoparticles. Green Chem 2006;8(1):34-38.##Andrews JM. Determination of minimum inhibitory concentrations. J Antimicrob Chemother 2001;48(Suppl 1):5-16.##Oblinger JL, Koburger JA. Understanding and teaching the most probable number technique. J Milk Food Technol 1975:38:540-545.##Shakibaie M, Forootanfar H, Mollazadeh-Moghaddam K, Bagherzadeh Z, Nafissi-Varcheh N, Shahverdi AR, et al. Green synthesis of gold nanoparticles by the marine microalga Tetraselmis suecica. Biotechnol Appl Biochem 2010;57(2):71-75.##Henglein A. Physicochemical properties of small metal particles in solution: “microelectrode” reactions, chemisorption, composite metal particles, and the atom-to-metal transition. J Phys Chem B 1993;97(21):5457-5471.##Sastry M, Mayya KS, Bandyopadhyay K. pH dependent changes in the optical properties of carboxylic acid derivatized silver colloidal particles. Colloid Surf A 1997;127(1-3):221-228.##Singaravelu G, Arockiamary J S, Kumarb VG, Govindaraju K. Colloid Surf B Biointerfaces 2007; 57: 97-101.##Cardoso VH, Goncalves DL, Angioletto E, Dal-Pizzol F, Streck EL. Microwave disinfection of gauze contaminated with bacteria and fungi. Indian J Med Microb 2007;25(4):428-429.##Pissuwan D, Cortie CH, Valenzuela SM, Cortie M. Functionalized gold nanoparticles for controlling pathogenic bacteria. Trends Biotechnol 2010;28(4): 207-213.##

Fetal Sex Determination using Non-Invasive Method of Cell-free Fetal DNA in Maternal Plasma of Pregnant Women During 6th– 10th Weeks of Gestation 2 2 In previous years, identification of fetal cells in maternal blood circulation has caused a new revolution in non-invasive method of prenatal diagnosis. Low number of fetal cells in maternal blood and long-term survival after pregnancy limited the use of fetal cells in diagnostic and clinical applications. With the discovery of cell-free fetal DNA (cffDNA) in plasma of pregnant women, access to genetic material of the fetus had become possible to determine early gender of a fetus in pregnancies at the risk of X-linked genetic conditions instead of applying invasive methods. Therefore in this study, the probability of detecting sequences on the Y chromosome in pregnant women has been evaluated to identify the gender of fetuses. Peripheral blood samples were obtained from 80 pregnant women at 6th to 10th weeks of gestation and then the fetal DNA was extracted from the plasma. Nested PCR was applied to detect the sequences of single copy SRY gene and multi copy DYS14 & DAZ genes on the Y chromosome of the male fetuses. At the end, all the obtained results were compared with the actual gender of the newborns. In 40 out of 42 born baby boys, the relevant gene sequences were identified and 95.2% sensitivity was obtained. Conclusion: Non-invasive early determination of fetal gender using cffDNA could be employed as a pre-test in the shortest possible time and with a high reliability to avoid applying invasive methods in cases where a fetus is at the risk of genetic diseases. 201 206 Maryam Zargari Biology Department, Science and Research Branch, Islamic Azad University (IAU) Biology Department, Science and Research Branch, Islamic Azad University (IAU) Iran Mohammad Reza Sadeghi Reproductive Biotechnology Research Centre, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Centre, Avicenna Research Institute, ACECR Iran Mohammad Hassan Shahhosseiny Microbiology Department, Shahr-e-Qods Branch, Islamic Azad University (IAU) Microbiology Department, Shahr-e-Qods Branch, Islamic Azad University (IAU) Iran Koorosh Kamali Reproductive Biotechnology Research Centre, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Centre, Avicenna Research Institute, ACECR Iran Kyomars Saliminejad Reproductive Biotechnology Research Centre, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Centre, Avicenna Research Institute, ACECR Iran Ali Esmaeilzadeh Reproductive Biotechnology Research Centre, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Centre, Avicenna Research Institute, ACECR Iran Hamid Reza Khorram Khorshid Reproductive Biotechnology Research Centre, Avicenna Research Institute, ACECRGenetic Research Centre, University of Social Welfare and Rehabilitation Sciences Reproductive Biotechnology Research Centre, Avicenna Research Institute, ACECRGenetic Research Centre, University of Social Welfare and Rehabilitation Sciences IranIran FetusGenetic materialPrenatal diagnosisSex determination 72.pdf Brandenburg H, Jahoda MG, Pijpers L, Reuss A, Kleyer WJ, Wladimiroff JW. Fetal loss rate after chorionic villus sampling and subsequent amniocentesis. Am J Med Genet 1990;35(2):178-180.##Odeh M, Granin V, Kais M, Ophir E, Bornstein J. Sonographic fetal sex determination. Obstet Gynecol Surv 2009;64(1):50-57.##Simpson JL, Elias S. Isolating fetal cells in maternal circulation for prenatal diagnosis. Prenat Diagn 1994;14(13):1229-1242.##Bianchi DW, Flint AF, Pizzimenti MF, Knoll JH, Latt SA. Isolation of fetal DNA from nucleated erythrocytes in maternal blood. Proc Natl Acad Sci USA 1990;87(9):3279-3283.##Hamada H, Arinami T, Kubo T, Hamaguchi H, Iwasaki H. Fetal nucleated cells in maternal peripheral blood: frequency and relationship to gestational age. Hum Genet 1993;91(5):427-432.##Hamada H, Arinami T, Hamaguchi H, Kubo T. Fetal nucleated cells in maternal peripheral blood after delivery. Am J Obstet Gynecol 1994;170(4): 1188-1193.##Chen XQ, Stroun M, Magnenat JL, Nicod LP, Kurt AM, Lyautey J, et al. Microsatellite alterations in plasma DNA of small cell lung cancer patients. Nat Med 1996;2(9):1033-1035.##Alberry M, Maddocks D, Jones M, Abdel Hadi M, Abdel-Fattah S, Avent N, et al. Free fetal DNA in maternal plasma in anembryonic pregnancies: confirmation that the origin is the trophoblast. Prenat Diagn 2007;27(5):415-418.##Lo YM, Corbetta N, Chamberlain PF, Rai V, Sargent IL, Redman CW, et al. Presence of fetal DNA in maternal plasma and serum. Lancet 1997;350(9076):485-487.##Lo YM, Tein MS, Lau TK, Haines CJ, Leung TN, Poon PM, et al. Quantitative analysis of fetal DNA in maternal plasma and serum: implications for non invasive prenatal diagnosis. Am J Hum Genet 1998;62(4):768-775.##Lo YM, Zhang J, Leung TN, Lau TK, Chang AM, Hjelm NM. Rapid clearance of fetal DNA from maternal plasma. American journal of human genetics. Am J Hum Genet 1999;64(1):218-224.##Li Y, Zimmermann B, Rusterholz C, Kang A, Holzgreve W, Hahn S. Size separation of circulatory DNA in maternal plasma permits ready detection of fetal DNA polymorphisms. Clin Chem 2004;50(6):1002-1011.##Wright CF, Burton H. The use of cell-free fetal nucleic acids in maternal blood for non-invasive prenatal diagnosis. Hum Reprod Update 2009;15(1):139-151.##Al-Yatama MK, Mustafa AS, Ali S, Abraham S, Khan Z, Khaja N. Detection of Y chromosome-specific DNA in the plasma and urine of pregnant women using nested polymerase chain reaction. Prenat Diagn 2001;21(5):399-402.##Hong P, Zhu PY, Huang YF, Luan JF. Study on fetal SRY gene in maternal plasma using nested polymerase chain reaction. Zhonghua Nan Ke Xue 2006;12(4):333-336.##Smid M, Lagona F, de Benassuti L, Ferrari A, Ferrari M, Cremonesi L. Evaluation of different approaches for fetal DNA analysis from maternal plasma and nucleated blood cells. Clin Chem 1999;45(9):1570-1572.##Zolotukhina TV, Shilova NV, Voskoboeva EY. Analysis of cell-free fetal DNA in plasma and serum of pregnant women. J Histochem Cytochem 2005;53(3):297-299.##Horinek A, Korabecna M, Panczak A, Gallova ZU, Nouzova K, Calda P, et al. Cell-free fetal DNA in maternal plasma during physiological single male pregnancies: Methodology issues and kinetics. Fetal Diagn Ther 2008;24(1):15-21.##Landy HJ, Keith LG. Hum Reprod Update 1998;4(2):177-183.##Viera AJ, Garrett JM. Understanding interobserver agreement: the kappa statistic. Fam Med 2005;37(5):360-363.##

Construction and Evaluation of an Expression Vector Containing Mtb32C (Rv0125) of Mycobacterium tuberculosis 2 2 Expressions of recombinant proteins for different applications are important objectives in molecular biotechnology; however, expression of some recombinant proteins is difficult. Several methods have been designed for expression of these proteins. The aim of this study was to construct a vector containing Mtb32C fragment of Mycobacterium tuberculosis (M.tuberculosis) as a fusion partner in order to improve the expression of fused recombinant proteins. Mtb32C was amplified by polymerase chain reaction (PCR). The amplified fragment was ligated into pET21b+ vector. Colony-PCR, enzyme digestion and DNA sequencing methods were used to confirm the recombinant vector. Colony-PCR showed a 420 bp fragment in size corresponding to the correct size of our fragment. In addition the recombinant plasmids sequencing showed the accuracy of the cloned fragment. For confirming the expression, reverse transcriptase (RT)-PCR analysis was performed showing a 420 bp fragment in agarose gel electrophoresis using specific primers. The construction of a vector containing Mtb32C fragment is promising as a fusion partner for future studies as it affected the expression of the fused proteins and increased immune responses against the partner. 207 210 Maryam Sadat Nabavinia Microbiology and Virology Research Center, Avicenna Research Institute & Department of Medical Bacteriology & Virology, Faculty of Medicine, Mashhad University of Medical Sciences Microbiology and Virology Research Center, Avicenna Research Institute & Department of Medical Bacteriology & Virology, Faculty of Medicine, Mashhad University of Medical Sciences Iran Mahboobeh Naderi Nasab Microbiology and Virology Research Center, Avicenna Research Institute & Department of Medical Bacteriology & Virology, Faculty of Medicine, Mashhad University of Medical Sciences Microbiology and Virology Research Center, Avicenna Research Institute & Department of Medical Bacteriology & Virology, Faculty of Medicine, Mashhad University of Medical Sciences Iran Zahra Meshkat Microbiology and Virology Research Center, Avicenna Research Institute & Department of Medical Bacteriology & Virology, Faculty of Medicine, Mashhad University of Medical SciencesWomen's Health Research Center, Mashhad University of Medical Sciences Microbiology and Virology Research Center, Avicenna Research Institute & Department of Medical Bacteriology & Virology, Faculty of Medicine, Mashhad University of Medical SciencesWomen's Health Research Center, Mashhad University of Medical Sciences IranIran Mohammad Derakhshan Microbiology and Virology Research Center, Avicenna Research Institute & Department of Medical Bacteriology & Virology, Faculty of Medicine, Mashhad University of Medical Sciences Microbiology and Virology Research Center, Avicenna Research Institute & Department of Medical Bacteriology & Virology, Faculty of Medicine, Mashhad University of Medical Sciences Iran Mehrangiz Khaje-Karamadini Women's Health Research Center, Mashhad University of Medical Sciences Women's Health Research Center, Mashhad University of Medical Sciences Iran Cloning vectorFusion partnerMolecular biotechnologyMycobacterium tuberculosisRecombinant proteins 74.pdf Meshkat Z. Construction of an expression vector containing immunogenic region of human papillomavirus type 16 E7 and HSP70 genes and evaluate the CMI responses in BALB/c mice. [dissertation]. [Tehran]: Tarbiat Modares University; 2007. 215p.##Esposito D, Chatterjee DK. Enhancement of soluble protein expression through the use of fusion tags. Curr Opin Biotechnol 2006;17(4):353-358.##Cabrita LD, Dai W, Bottomley SP. A family of E. coli expression vectors for laboratory scale and high throughput soluble protein production. BMC Biotechnol 2006;6(1):12.##Baneyx F, Mujacic M. Recombinant protein folding and misfolding in Escherichia coli. Nature Biotechnol 2004;22(11):1399-1408.##Cao H. Expression, purification, and biochemical characterization of the anti inflammatory tristetraprolin: a zinc-dependent mRNA binding protein affected by posttranslational modifications. Biochemistry 2004;43(43):13724-13738. ##Hannig G, Makrides SC. 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