Editorial 2 2 In the previous editorial I made mention of the two important conferences on ethical aspects of genetic technologies that had been planned to be held in Iran (November 2010 and February 2011). In the November conference which was held by the Avicenna Research Institute, the ethical and social aspects of using genetic technologies were presented to the academic and non-academic audiences. This conference was widely covered by the national media and the subject of genetics with its interdisciplinary nature initiated discussion between experts in both fields of natural and social sciences. Since some of our colleagues were not able to attend the conference, I thought it would be appropriate to provide a brief summary of the theme of presentations in the conference. The two day conference was organized in four panels and the topics presented in each panel were discussed from ethical, legal and social points of view. The four panels were as follows: 1) Genetics and the Emergence of Life 2) Genetics in Diagnosis and Treatment of Diseases 3) Plant and Animal Genetics 4) New Findings on the Science of Genetics In the first panel 10 lectures were presented on topics of eugenics, cloning, genetic diagnostics and abortion. In the second panel 7 lectures were presented on topics of stem cells, Pre-implantation Genetic Diagnosis (PGD), pharmacogenomics and gene therapy. In the third panel on the second day, 8 lectures were presented on topics of genetically modified organisms (GMO), transgenic plants, bio-safety issues and its implementations at the national level, agricultural genetic engineering, Intellectual Property, issues of patent and bio-safety and the rights of consumers with regards to transgenic products. In the fourth panel on the second day, 9 lectures were presented on topics of mental health and human genome, genetic enhancements, completion of human genome and social implications, genetic predisposition to crime and legal responsibility, DNA tests and proof of identity. The topics listed above clearly demonstrate that genetic engineering has had a profound impact on human life in societies worldwide at multilevel. Therefore, it is important for leaders of societies in the modern world to pay attention to the advances in genetic technologies and prepare themselves and institutions under their control to face the challenges which these new technologies induce in the areas of ethics, law and social policies. 160 160 Ali M. Ardekani Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Editorial 160.pdf ####

The Role of MicroRNAs in Human Diseases 2 2 MicroRNAs (miRNAs) are short RNA molecules which bind to target mRNAs, resulting in translational repression and gene silencing and are found in all eukaryotic cells. Approximately 2200 miRNA genes have been reported to exist in the mammalian genome, from which over 1000 belong to the human genome. Many major cellular functions such as development, differentiation, growth, and metabolism are known to be regulated by miRNAs. Proximity to other genes in the genome and their locations in introns of coding genes, noncoding genes and exons have been reported to have a major influence on the level of gene expressions in eukaryotic cells. miRNAs are well conserved in eukaryotic system and are believed to be an essential and evolutionary ancient component of gene regulatory networks. 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Regulation Studies of Telomerase Gene in Cancer Cells by Lentinan 2 2 Lentinan a polysaccharide from medicinal mushroom i.e Lentinus, has been known to have anticancer properties. Telomerase activity is not observed in normal healthy cells, whereas in cancerous cells telomerase expression is high. Telomerase represents a promising cancer therapeutic target. We investigated the inhibitory effect of lentinan on telomerase reverse transcriptase gene (hTERT) which is essential for telomerase activity. To assess the transcriptional effect, DLD -1 cancer cells were cultured in the presence of various concentrations of lentinan. TRAP assay, RT-PCR analysis were performed to find telomerase activity and hTERT gene expression respectively. Since C-myc is known to regulate hTERT, expression of C-myc was also determined. Culturing cells with lentinan resulted in down regulation of hTERT and C-myc expression.These results indicate that lentinan inhibits telomerase activity by down regulating hTERT expression via suppression of C-myc in cancer cells. 181 186 Kamma Sreenivasulu Department of Biotechnology, KL University Department of Biotechnology, KL University India Muvva Vijayalakshmi Department of Botany and Microbiology, Acharya Nagarjuna University Department of Botany and Microbiology, Acharya Nagarjuna University India Krothapalli RS. Sambasivarao Department of Biotechnology, Acharya Nagarjuna University Department of Biotechnology, Acharya Nagarjuna University India C-myc geneGene expressionhTERTLentinanTelomerase 45.pdf Morin GB. The human telomere terminal transferase enzyme is a ribo nucleoprotein that synthesizes TTAGGG repeats. Cell 1989;59(3):521-529. ##Kim NW, Piatyszek MA, Prowse KR, Harley CB, West MD, HO PLC, et al. Specific association of human telomerase activity with immortal cells and cancers. Science 1994;266:2011-2015. ##Nakamura TM, Morin GB,Chapman KB, Weinrich SL, Andrews WH, Lingner J, et al. Telomerase catalytic subunit homolgs from fission yeast and human. Science 1977; 277(5328):955-959. ##Wu KJ, Grandori C, Amacker M, Simon-Vermot N, Polack A, Lingner J, et al. Direct activation of TERT transcription by C-myc. Nat Genet 1999;21: 220-224. ##Eitsuka T, Nakagawa K, Suzuki T, Miyazawa T. Polyunsaturated fatty acids inhibit telomerase activity in DLD-1 human colorectal adenocarcinoma cells: a dual mechanism approach. Biochim Biophys Acta 2005;1737(1):1-10. ##Biray Avci C, Dogan ZO, Yilmaz S, Numanoglu S, Topçuoglu N, Gündüz C. Effect of resveratrol and caffeic acid phenethyl ester on the expressions of p53, MDM2, PIK3CA and hTERT in human breast cancer cell line. Adv Mol Med 2007;3(1):45-48. ##Elliott PJ, Jirousek M. Sirtuins: novel targets for metabolic disease. Curr Opin Investig Drugs 2008;9(4):371-378. ##Hideshima T, Chauhan D, Shima Y, Raje N, Davies FE, Tai YT, et al. Thalidomide and its analogs overcome drug resistance of human multiple myeloma cells to conventional therapy. Blood 2000;96(9):2943–2950. ##Pitts AE, Corey DR. Inhibition of human telomerase by 2’-0’-mehyl RNA. Proc Natl Acad Sci USA 1998;95(20):11549–11554. ##Strahl C, Blackburn EH. Effects of reverse transcriptase inhibitors on telomere length and telomerase activity in two immortalized cell lines. Mol Cell Biol 1996;16(1):53-65. ##K Damm, Hemmann U, Garin-Chesa P, Hauel N, Kauffmann I, Priepke H, et al. A highly selective telomerase inhibitor limiting human cancer cell proliferation. EMBO J 2001;20(24):6958–6968. ##Meyerson M, Counter CM, Eaton EN, Ellisen LW, Steiner P, Caddle SD, et al. hEST2, the putative human telomerase catalytic subunit gene, is up-regulated in tumor cells and during immortalization. Cell 1997;90(4):785-795. ##Hisatake J, Kubota T, Hisatake Y, Uskokovic M, Tomoyasu S, Koeffler HP. 5,6-trans -16-ene-Vitamin D3: a new class of potent inhibitors of proliferation of prostate, breast, and myeloid leukemic cells. Cancer Res 1999;59(16):4023-4029. ##Ogretmen B, Kraveka JM, Schandy D, Usta J, Hannum YA, Obeid LM. Molecular mechanisms of ceramide-mediated telomerase inhibition in the A249 human lung adenocarcinoma cell line. J Biol Chem 2001;276(35):32506-32514. ##Ramachandran C, Fonseca HB, Jhabvala P, Escalon EA, Melnick SJ. Curcumin inhibits telomerase activity through human telomerase reverse transcriptase in MCF-7 breast cancer cell line. Cancer Lett 2002;184(1):1-6. ##Takakura M, Kyo S, Kanaya T, Tanaka M, Inoue M. Expression of human telomerase subunits and correlation with telomerase activity in cervical cancer. Cancer Res 1998;58(7):1558-1561. ##

Cytotoxic Activities of Silver Nanoparticles and Silver Ions in Parent and Tamoxifen-Resistant T47D Human Breast Cancer Cells and Their Combination Effects with Tamoxifen against Resistant Cells 2 2 Studies on biomedical applications of nanoparticles are growing with a rapid pace. In medicine, nanoparticles may be the solution for multi-drug-resistance which is still a major drawback in chemotherapy of cancer. In the present study, we investigated the potential cytotoxic effect of silver nanoparticles (Ag NPs) and silver ions (Ag+) in both parent and tamoxifen-resistant T47D cells in presence and absence of tamoxifen. Ag NPs were synthesized (< 28 nm) and MTT assay was carried out. The associated IC50 values were found to be: 6.31 ?g/ml for Ag NPs/parent cells, 37.06 ?g/ml for Ag NPs/tamoxifen-resistant cells, 33.06 ?g/ml for Ag+/parent cells and 10.10 ?g/ml for Ag+/resistant cells. As a separate experiment, the effect of subinhibitory concentrations of Ag NPs and Ag+ on the proliferation of tamoxifen resistant cells was evaluated at non-toxic concentrations of tamoxifen. Our results suggested that in non-cytotoxic concentrations of silver nanomaterials and tamoxifen, the combinations of Ag+-tamoxifen and Ag NPs-tamoxifen are still cytotoxic. This finding may be of great potential benefit in chemotherapy of breast cancer; since much lower doses of tamoxifen may be needed to produce the same cytotoxic effect and side effects will be reduced. 187 196 Seyed Naser Ostad Department of Pharmacology and Toxicology, Faculty of Pharmacy, Tehran University of Medical Sciences Department of Pharmacology and Toxicology, Faculty of Pharmacy, Tehran University of Medical Sciences Iran Shahrzad Dehnad Department of Pharmaceutical Biotechnology, Biotechnology Research Center, Faculty of Pharmacy, Tehran University of Medical Sciences Pharmaceutical Research Division, Azad Islamic University Department of Pharmaceutical Biotechnology, Biotechnology Research Center, Faculty of Pharmacy, Tehran University of Medical Sciences Pharmaceutical Research Division, Azad Islamic University IranIran Zeinab Esmail Nazari Department of Pharmaceutical Biotechnology, Biotechnology Research Center, Faculty of Pharmacy, Tehran University of Medical Sciences Department of Pharmaceutical Biotechnology, Biotechnology Research Center, Faculty of Pharmacy, Tehran University of Medical Sciences Iran Shohreh Tavajohi Fini Department of Pharmacology and Toxicology, Faculty of Pharmacy, Tehran University of Medical Sciences Department of Pharmacology and Toxicology, Faculty of Pharmacy, Tehran University of Medical Sciences Iran Narges Mokhtari Department of Pharmaceutical Biotechnology, Biotechnology Research Center, Faculty of Pharmacy, Tehran University of Medical Sciences Department of Pharmaceutical Biotechnology, Biotechnology Research Center, Faculty of Pharmacy, Tehran University of Medical Sciences Iran Mojtaba Shakibaie Department of Pharmaceutical Biotechnology, Biotechnology Research Center, Faculty of Pharmacy, Tehran University of Medical Sciences Department of Pharmaceutical Biotechnology, Biotechnology Research Center, Faculty of Pharmacy, Tehran University of Medical Sciences Iran Ahmad Reza Shahverdi Department of Pharmaceutical Biotechnology, Biotechnology Research Center, Faculty of Pharmacy, Tehran University of Medical Sciences Department of Pharmaceutical Biotechnology, Biotechnology Research Center, Faculty of Pharmacy, Tehran University of Medical Sciences Iran Breast neoplasmsChemotherapyCytotoxicityNanoparticlesTamoxifen 46.pdf Parak WJ, Manna L, Simmel FC, Gerion D, Alivisatos P. 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High Expression of Methylotrophic Yeast-Derived Recombinant Human Erythropoietin in a pH-Controlled Batch System 2 2 To accomplish the worldwide demand for recombinant human erythropoietin (rHuEpo) as a therapeutic, application of cost-efficient expression system of methylotrophic yeast Pichia pastoris (P. pastoris) rather than mammalian cells is indispensable. Herein, a report on high levels secreted-expression of Pichia-derived rHuEpo by batch fermentation in a pH stabilized format is presented. The full length cDNA of rHuEpo was inserted into pPICZaA vector under control of AOX1 promoter, downstream of the secretion-a-factor and electroporated into P. pastoris strain X33. The highest expression transformant was selected by screening among the colonies surviving high concentration of Zeocin (1.0 mg/ml), followed by comparative small scale expression analysis by ELISA. Stabilization of pH around 6.0 by adding phosphoric acid into the culture media during induction period, improved the yield of expression to 150 mg/l of the media. Single-step Nickel-affinity chromatography was employed for purification of rHuEpo-6xHis to 80% purity. Analyses by SDS- PAGE, Western blot and N-terminal protein sequencing confirmed the authenticity of the 33 kDa expressed rHuEpo with a native N-terminal indicating the proper cleavage of secretion-signal. Results of this study, further confirmed the possibility of employing methylotrophic yeast for scaled up production aims of rHuEpo as a cost-efficient expression system when provided evidence for higher expression yields through application of pH-controlled systems. 197 206 Ahmad Maleki Faculty of Pharmacy, Tehran University of Medical ScienceResearch and Production Plant, Pasteur Institute of Iran Faculty of Pharmacy, Tehran University of Medical Science IranIran Farzin Roohvand Hepatitis & AIDS Department, Pasteur Institute of Iran Iran Hosnieh Tajerzadeh Faculty of Pharmacy, Tehran University of Medical Science Faculty of Pharmacy, Tehran University of Medical Science Iran Hossein Khanahmad Research and Production plant, Pasteur Institute of Iran Iran Maryam B. Nobari Open University of Zanjan Open University of Zanjan Iran Ahmad Beiruti Research and Production plant, Pasteur Institute of Iran Iran Abdolhossein Rouholamini Najafabadi Faculty of Pharmacy, Tehran University of Medical Science Faculty of Pharmacy, Tehran University of Medical Science Iran ErythropoietinFermentationPichia PastorisYeasts 47.pdf Egrie J. The cloning and production of recombinant human erythropoietin. Pharmacotherapy 1990;10 (Pt 2):3S-8S. ##Celik E, Calik P, Halloran SM, Oliver SG. Production of recombinant human erythropoietin from Pichia pastoris and its structural analysis. J Appl Microbiol 2007;103(6):2084-2094. ##Elliott S, Pham E, Macdougall IC. Erythropoietins: a common mechanism of action. Exp Hematol 2008;36(12):1573-84. ##Lee-Huang S. Cloning and expression of human erythropoietin cDNA in Escherichia coli. Proc Natl Acad Sci USA 1984;81(9):2708-2712. ##Nagao M, Inoue K, Moon SK, Masuda S, Takagi H, Udaka S, et al. Secretory production of erythropoietin and the extracellular domain of the erythropoietin receptor by Bacillus brevis: affinity purification and characterization. Biosci Biotechnol Biochem 1997;61(4):670-674. ##Elliott S, Giffin J, Suggs S, Lau EP, Banks AR. Secretion of glycosylated human erythropoietin from yeast directed by the alpha-factor leader region. Gene 1989;79(1):167-180. ##Kim YK, Shin HS, Tomiya N, Lee YC, Betenbaugh MJ, Cha HJ. Production and N-glycan analysis of secreted human erythropoietin glycoprotein in stably transfected Drosophila S2 cells. Biotechnol Bioeng 2005;92(4):452-461. ##Takeuchi M, Takasaki S, Shimada M, Kobata A. Role of sugar chains in the in vitro biological activity of human erythropoietin produced in recombinant Chinese hamster ovary cells. J Biol Chem 1990;265: 12127-12130. ##Kjeldsen T. Yeast secretory expression of insulin precursors. Appl Microbiol Biotechnol 2000;54(3): 277-286. ##Gellissen G. Heterologous protein production in methylotrophic yeasts. Appl Microbiol Biotechnol 2000;54(6):741-750. ##Hartner FS, Glieder A. Regulation of methanol utilisation pathway genes in yeasts. Microbial Cell Fact 2006;5:39. ##Yurimoto H, Sakai Y. Methanol-inducible gene expression and heterologous protein production in the methylotrophic yeast Candida boidinii. Biotechnol Appl Biochem 2009;53(Pt 2):85-92. ##Bollok M, Resina D, Valero F, Ferrer P. Recent patents on the Pichia pastoris expression system: expanding the toolbox for recombinant protein production. Recent Pat Biotechnol 2009;3(3):192-201. ##Daly R, Hearn MT. Expression of heterologous proteins in Pichia pastoris: a useful experimental tool in protein engineering and production. J Mol Recognit 2005;18(2):119-138. ##Li P, Anumanthan A, Gong Gao X, Ilangovan K, Suzara VV, Düzgünes N, et al. Expression of recombinant proteins in Pichia pastoris. Appl Biochem Biotechnol 2007;142(2):105-124. ##Celik E, Calik P, Oliver S. Fed-batch methanol feeding strategy for recombinant protein production by Pichia pastoris in the presence of co-substrate sorbitol. Yeast 2009;26(9):473-484. ##Bradford MM. A rapid and sensitive for the quantitation of microgram quantities of protein utilizing the principle of protein dye binding. Anal Biochem 1976;72(1-2):248-254. ##Laemmli UK. Cleavage of structural proteins during assembly of head of bacteriophage-T4. Nature 1970;227:680-685. ##Clare JJ, Romanos MA, Rayment FB, Rowedder JE, Smith MA, Payne MM, et al. Production of mouse epidermal growth factor in yeast: high-level secretion using Pichia pastoris strains containing multiple gene copies. Gene 1991;105(2):205-212. ##Couderc R, Baratti J. Oxidation of methanol by the yeast, Pichia pastoris. Purification and properties of the alcohol oxidase. Agric Biol Chem 1980;44: 2279-2289. ##Apte-Deshpande A, Somani S, Mandal G, Soorapaneni S, Padmanabhan S. Over expression and analysis of O-glycosylated recombinant human granulocyte colony stimulating factor in Pichia pastoris using Agilent 2100 Bioanalyzer. J Biotechnol 2009;143(1):44-50. ##Cregg JM, Cereghino JL, Shi J, Higgins DR. Recombinant protein expression in Pichia pastoris. Mol Biotechnol 2000;16(1):23-52. ##Su M, Xu T, Wang D, Zhou Y, Niu C, Yan W. High yield and purification of recombinant human apolipoprotein E3 in Pichia pastoris. Protein Expr Purif 2009;68(1):7-11. ##Prevatt WD, Sreekrishna K, inventors; Research Corporation Technologies Inc., assignee. Expression of human serum albumin in Pichia pastoris. United States patent US 5,330,901. 1994 Jul 19. ##País-Chanfrau JM, García Y, Licor L, Besada V, Castellanos-Serra L, CI Cabello, et al. Improving the expression of mini-proinsulin in Pichia pastoris. Biotechnol Lett 2004;26(4):1269-1272. ##Liu R, Lin Q, Sun Y, Qiu Y, Xu T, Ding H, Guo X. Optimization of fermentation conditions for maximal recombinant hepatitis B surface antigen particle production in Pichia pastoris. Sheng Wu Gong Cheng Xue Bao 2008;24(12):2098-2105. ##Lin H, Kim T, Xiong F, Yang X. Enhancing the production of Fc fusion protein in fed-batch fermentation of Pichia pastoris by design of experiments. Biotechnol Prog 2007;23(3):621-625. ##Vellanki RN, Potumarthi R, Mangamoori LN. Constitutive expression and optimization of nutrients for streptokinase production by Pichia pastoris using statistical methods. Appl Biochem Biotechnol 2009;158(1):25-40. ##Kukuruzinska MA, Bergh MLE, Jackson BJ. Protein glycosylation in yeast. Annu Rev Biochem 1987;56: 915-944. ##Zsebo KM, Lu HS, Fieschko JC, Goldstein L, Davis J, Duker K, et al. Protein secretion from Saccharomyces cerevisiae directed by the prepro-alpha-factor leader region. J Biol Chem 1986;261:5858-5865. ##Hamilton SR, Davidson RC, Sethuraman N, Nett JH, Jiang Y, Rios S, et al. 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Development of a Sensitive Enzyme-Linked Immunosorbent Assay for Detection of Hepatitis B Surface Antigen Using Novel Monoclonal Antibodies 2 2 Hepatitis B virus (HBV) infection is the 10th leading cause of death worldwide. The most important diagnostic and screening marker for HBV infection is Hepatitis B surface antigen (HBsAg), and the most widely used HBsAg screening test is Enzyme-linked Immunosorbent Assay (ELISA). In this study, an ELISA assay has been developed for detection of HBsAg using two novel monoclonal antibodies (mAb) as capture layer and a polyclonal biotinylated Ab as detector phase. We evaluated the sensitivity, specificity, detection limit, seroconversion time, positive and negative predictive values and reproducibility of our assay with standard panels and different serum samples. The results were compared with a well established commercial kit. Both assays showed similar detection limit values of 0.5 to 0.7 ng/ml and the same seroconversion periods of 42 and 65 days for “ad” and “ay” serotypes of HBsAg, respectively. Sensitivity and specificity of the assay were 98.98% and 99.6%, respectively. The positive and negative predictive values of our assay were also calculated as 99.49% and 99.2%, respectively. Analysis of reproducibility of the present assay demonstrated 3.96% and 5.85% intra-and inter-assay coefficient of variations, respectively, which were less than those obtained by the commercial kit. There was a highly significant correlation between our designed assay and the commercial ELISA kit (p < 0.0001, r = 0.957). Altogether, our results indicate that the designed assay is comparable to the commercial kit in terms of sensitivity, specificity, positive and negative predictive values and reproducibility and could be employed for diagnosis of HBV infection in blood samples. 207 214 Yaghoub Yazdani Department of Immunology, School of Public Health, Tehran University of Medical Sciences Department of Molecular Medicine, Faculty of Advanced Medical Science Technologies, Golestan University of Medical Sciences Department of Immunology, School of Public Health, Tehran University of Medical Sciences Department of Molecular Medicine, Faculty of Advanced Medical Science Technologies, Golestan University of Medical Sciences IranIran Azam Roohi Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Iran Jalal Khoshnoodi 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 Fazel Shokri Department of Immunology, School of Public Health, Tehran University of Medical Sciences Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Department of Immunology, School of Public Health, Tehran University of Medical Sciences Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR IranIran ELISAHepatitis BHepatitis B surface antigensMonoclonal antibody 48.pdf Chen DS. Toward elimination and eradication of hepatitis B. J Gastroenterol Hepatol 2010;25(1):19-25. ##Glebe D. Recent advances in hepatitis B virus research: A German point of view. World J Gastroenterol 2007;13(1):8-13. ##Yokosuka O, Arai M. Molecular biology of hepatitis B virus: effect of nucleotide substitutions on the clinical features of chronic hepatitis B. Med Mol Morphol 2006;39(3):113-120. ##Ganem D, Prince AM. Hepatitis B virus infection-natural history and clinical consequences. N Engl J Med 2004;350(11):1118-1129. ##Hatzakis A, Magiorkinis E, Haida C. HBV virological assessment. J Hepatol 2006;44(1):S71-S76. ##Pungpapong S, Kim WR, Poterucha JJ. Natural history of hepatitis B virus infection: An update for clinicians. Mayo Clinic Proc 2007;82(8):967-975. ##Urbani S, Fagnoni F, Missale G, Franchini M. The role of anti-core antibody response in the detection of occult hepatitis B virus infection. Clin Chem Lab Med 2010;48(1):23-29. ##Pawlotsky JM. Virologic techniques for the diagnosis and monitoring of hepatitis B. Gastroenterol Clin Biol 2008;32(1 Pt 2):S56-S63. ##Kuhns MC, Busch MP. New strategies for blood donor screening for hepatitis B virus - Nucleic acid testing versus immunoassay methods. Mol Diag Ther 2006;10(2):77-91. ##Niederhauser C, Mansouri Taleghani B, Graziani M, Stolz M, Tinguely C, Schneider P. Blood donor screening: how to decrease the risk of transfusion-transmitted hepatitis B virus? Swiss Med Wkly 2008;138(9-10):134-141. ##Raimondo G, Pollicino T, Cacciola I, Squadrito G. Occult hepatitis B virus infection. J Hepatol 2007;46(1):160-170. ##Vernet G. Molecular diagnostics in virology. J Clin Virol 2004; 31(4):239-247. ##Dwyer R. The ADVIA Centaur (R) infectious disease assays: a technical review. J Clin Virol 2004;30(1):S1-S5. ##Biswas R, Tabor E, Hsia CC, Wright DJ, Laycock ME, Fiebig EW et al. Comparative sensitivity of HBVNATs and HBsAg assays for detection of acute HBV infection. 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Trans R Soc Trop Med Hyg 2008;102(8):787-792. ##Borgniet O, Parvaz P, Bouix C, Chevallier P, Trepo C, Andre P, et al. Clearance of serum HBsAg and anti-HBs seroconversion following antiviral therapy for chronic hepatitis B. J Med Virol 2009;81(8):1336-1342. ##Lee JH, Kim SJ, Ahn SH, Lee J, Park Y, Kim HS. Correlation between quantitative serum HBsAg and HBV DNA test in Korean patients who showed high level of HBsAg. J Clin Pathol 2010; 63(11):1027-1031. ##Galli C, Orlandini E, Penzo L, Badiale R, Caltran G, Valverde S, et al. What is the role of serology for the study of chronic hepatitis B virus infection in the age of molecular biology? J Med Virol 2008; 80(6):974-979. ##Matsubara N, Kusano O, Sugamata Y, Itoh T, Mizuii M, Tanaka J, et al. A novel hepatitis B virus surface antigen immunoassay as sensitive as hepatitis B virus nucleic acid testing in detecting early infection. Transfusion 2009;49(3):585-595. ##Roohi A, Yazdani Y, Khoshnoodi J, Jazayeri SM, Carman WF, Chamankhah M, et al. Differential reactivity of mouse monoclonal anti-HBs antibodies with recombinant mutant HBs antigens. World J Gastroenterol 2006;12(33):5368-5374. ##Roohi A, Khoshnoodi J, Zarnani AH, Shokri F. Epitope mapping of recombinant hepatitis B surface antigen by murine monoclonal antibodies. Hybridoma 2005; 24(2):71-77. ##Growther J. The ELISA guidebook. New Jersey: Human Press Inc;2010. ##Pekary AE, Turner LF Jr, Hershman JM. New immunoenzymatic assay for human thyrotropin compared with two radioimmunoassays. Clin Chem 1986;32(3):511-514. ##Yang J, Kim JH, Kim Y. Comparison of nine different qualitative HBsAg assay kits. Korean J Lab Med 2010;30(2):178-184. ##Weber B, Bayer A, Kirch P, Schluter V, Schlieper D, Melchior W. Improved detection of hepatitis B virus surface antigen by a new rapid automated assay. J Clin Microbiol 1999;37(8):2639-2647. ##Ismail N, Fish GE, Smith MB. Laboratory evaluation of a fully automated chemiluminescence immunoassay for rapid detection of HBsAg, antibodies to HBsAg, and antibodies to hepatitis C virus. J Clin Microbiol 2004;42(2):610-617. ##Leyva A, Franco A, Gonzalez T, Sanchez JC, Lopez I, Geada D, et al. A rapid and sensitive ELISA to quantify an HBsAg specific monoclonal antibody and a plant-derived antibody during their downstream purification process. Biologicals 2007;35(1):19-25. ##Toplikar E, Carlomagno A, Rojkin LF, Gariglio R, Lorenzo LE. Development of an Enzyme-Immunoassay for the detection of Hepatitis-B Surface-Antigen employing monoclonal antibodies. J Clin Lab Anal 1993;7(6):324-328. ##

Hypolipidemic Activity of Chloroform Extract of Mimosa pudica Leaves 2 2 Mimosa pudica Lin., known as chue Mue, is a stout straggling prostrate shrubby plant, with spinous stipules and globose pinkish flower heads, and grows as weed in almost all parts of the country. It is traditionally used for its various properties and hence in the present study, chloroform extract of Mimosa pudica leaves has been screened for its hypolipidemic activity. Hypolipidemic activity is screened by inducing hyperlipidemia with the help of atherogenic diet in wistar albino rats and serum levels of various biochemical parameters such as total cholesterol, triglycerides, LDL, VLDL and HDL cholesterol were determined. Atherogenic index shows the measure of the atherogenic potential of the drugs. Chloroform extract showed significant (p < 0.05) hypolipidemic effect by lowering the serum levels of biochemical parameters such as significant reduction in the level of serum cholesterol, triglyceride, LDL, VLDL and increase in HDL level which was similar to the standard drug Atorvastatin. Chloroform extract exhibited significant atherogenic index and percentage protection against hyperlipidemia. These biochemical observations were in turn confirmed by histopathological examinations of aorta, liver and kidney sections and are comparable with the standard hypolipidemic drug Atorvastatin. Preliminary phytochemical analysis revealed the presence of phytoconstituents such as steroids, flavonoids, glycosides, alkaloids, phenolic compounds which is further confirmed by the thin layer chromatography, High Performance Thin Layer Chromatography (HPTLC). The overall experimental results suggests that the biologically active phytoconstituents such as flavonoids, glycosides alkaloids present in the chloroform extract of Mimosa pudica, may be responsible for the significant hypolipidemic activity and the results justify the use of Mimosa pudica as a significant hypolipidemic agent. 215 222 Rekha Rajendran Department of Pharmacognosy and Phytochemistry, SRM Univeristy, SRM College of Pharmacy Department of Pharmacognosy and Phytochemistry, SRM Univeristy, SRM College of Pharmacy India Ekambaram Krishnakumar Pharmaceutical Biotechnology, Mohamed Sathak A. J. College of Pharmacy India Atherogenic dietAtorvastatinBiological markers ChloroformThin layer chromatography 49.pdf Ghani A. Medicinal plants of Bangladesh with chemical constituents and uses. 2nd ed. Dhaka: Asiatic Society of Bangladesh; 1998. ##Umamaheswari S, Prince PS. Antihyperglycemic effect of ilogen excel, an ayurvedic herbal formulation in streptozotocin-induced diabetes mellitus. Acta Pol Pharm 2007;64(1):53-61. ##Balakrishnan N, Suresh D, Pandian GS, Edwin E, Sheeja E. Anti-diarrhoeal potential of Mimosa pudica root extracts. Indian J Nat Prod 2006;22(2):21-23. ##Bum EN, Dawack DL, Schmutz M, Rakotonirina A, Rakotonirina SV, Portet C, et al. Anticonvulsant activity of Mimosa pudia decoction. Fitoterapia 2004;75(3-4):309-314. ##Chowdhury SA, Islam J, Rahaman M, Rahman M, Rumzhum NN, Sultana R, et al. Cytotoxic, anti-microbial and anti-oxidant activities of the different plant parts of Mimosa pudica. S J Pharm Sci 2008;1(1&2):80-84. ##Rajendran R, Hemalatha S, Akasakalai K, MadhuKrishna CH, Sohil B, Sundaram V, et al. Hepatoprotective activity of Mimosa pudica leaves against Carbon tetrachloride induced toxicity. J Nat Prod 2009;2:116-122. ##Manrique CM, Rosenzweig JL, Umpierrez GE. Patient information page from the hormone foundation: diabetes, dyslipidemia and heart protection. J Clin Endocrinol Metab 2009;94(1):0. ##Xu Y, He Z, King GL. Introduction of hyperglycemia and dyslipidemia in the pathogenesis of diabetic vascular complications. Curr Diab Rep 2005;5(2):91-97. ##Kokate CK. Practical pharmacognosy. 4th ed. New Delhi: Vallabh Prakashan; 1993. ##Harborne JB. Phytochemical methods: a guide to modern technique of plant analysis. 2nd ed. New York: Chapman and Hall; 1984. ##Wagner G, Bladt S. Plant drug analysis: A thin layer chromatography atlas. 2nd ed. Heidelberg: Springer; 1996. ##Ecobichon DJ. The basis of toxicology testing. 2nd ed. New York: CRC press; 1997. ##Ahire AE, Laddha KS. Hypolipidemic effects of Carthamus tinctorius in rats. Indian Drugs 2005;42(8):545-546. ##Arad Y, Ramakrishna R, Ginsberg HN. Effect of lovastatin on very-low-density lipoprotein triglyceride metabolism in subjects with combined hyperlipidemia; evidence for reduced assembly and secretion of triglyceride rich lipoprotein. Metabolism 1992;41(5):487-493. ##Rajendran R, Hemachander R, Ezhilarasan T, Keerthana C, Saroja DL, Saichand KV, et al. Phytochemical analysis and in-vitro antioxidant activity of Mimosa pudica Lin. leaves. Research J Pharm and Tech 2010;3(2):551-555. ##Witzum JL. The oxidation hypothesis of atherosclerosis. Lancet 1994;344(8925):793-795. ##Alexander RW. Hypertension and pathogenesis of atherosclerosis - oxidative stress and mediation of arterial inflammatory response - A new perspective. Hypertension 1995;25:155-161. ##Steinberg D, Gotto AM. Preventing coronary artery disease by lowering cholesterol levels- Fifty years from bench to bedside. JAMA 1999;282(21):2043-2050. ##Fernandes NP, Lagishetty CV, Panda VS, Naik SR. An experimental evaluation of the antidiabetic and antilipidemic properties of a standardized Momordica charantia fruit extract. BMC Complement Altern Med 2007;7:29-37. ##Saravanan R, Pari L. Antihyperlipidemic and antiperoxidative effect of Diasulin, a polyherbal formulation in alloxan induced hyperglycemic rats. BMC Complement Altern Med 2005;5:14-23. ##

A Brief View on Molecular Diagnosis and Surveillance of West Nile Virus 2 2 <p> West Nile Virus (WNV) is an important zoo-notic agent having a wide host range. Due to its emergence with increased virulence in a wide geographical range, its monitoring becomes im-perative. Development of more rapid and sensitive molecular techniques for instance Reverse Trans-criptase-Polymerase Chain Reaction (RT-PCR), reverse transcription loop-mediated isothermal amplification (RT-LAMP) and Nucleic Acid Se-quence Based Amplification (NASBA) assays are vital for detection of the virus. Various surveil-lance techniques according to epidemiological, climatic and geographical conditions in the ex-posed area have also been developed. The surveil-lance can be set up at different levels of the WNV transmission cycle using birds, horses and mos-quitoes as sentinels.</p> 223 225 Pranay Kumar Department of Veterinary Microbiology, College of Veterinary Sciences & A. H. India Shanker K.Singh Division of Medicine, Indian Veterinary Research Institute India Yogranjan R.Singh Department of Agriculture Biotechnology, College of Agriculture India Mayurdhvaj K.Jhala Department of Veterinary Microbiology, College of Veterinary Sciences & A. H. India Editorial 50.pdf Beasley DWC, Li L, Suderman MT, Barrett ADT. Mouse neuroinvasive phenotype of West Nile virus strains varies depending upon virus genotype. Virology 2002;296(1):17-23. ##CDC. West Nile Virus activity — United States, 2009. Morbid Mortal Wkly Rep 2010;59(25):769-772. ##Artsob H, Gubler DJ, Enria DA, Morales MA, Pupo M, Bunning ML, et al. West Nile virus in the new world: trends in the spread and proliferation of West Nile virus in the western hemisphere. Zoonoses Public Health 2009;56(6-7):357-369. ##Venter M, Steyl J, Human S, Weyer J, Zaayman D, Blumberg L, et al. Transmission of West Nile virus during horse autopsy. Emerg Infect Dis 2010;16 (3):573-575. ##Hukkanen RR, Liggitt HD, Kelley ST, Grant R, Anderson D, Beaty BJ, et al. Comparison of commercially available and novel West Nile virus immunoassays for detection of seroconversion in pig-tailed macaques (Macaca nemestrina). Comp Med 2006;56(1):46-54. ##Wong SJ, Demarest VL, Boyle RH, Wang T, Ledizet M, Kar K, et al. Detection of human anti-flavivirus antibodies with a west nile virus recombinant antigen microsphere immunoassay. J Clin Microbiol 2004;42(1):65-67. ##Naze F, Le Roux K, Schuffenecker I, Zeller H, Staikowsky F, Grivard P, et al. Simultaneous detection and quantitation of Chikungunya, dengue and West Nile viruses by multiplex RT-PCR assays and Dengue virus typing using high resolution melting. J Virol Methods 2009;162(1-2):1-7. ##Zaayman D, Human S, Venter M. A highly sensitive method for the detection and genotyping of West Nile virus by real-time PCR. J Virol Methods 2009;157(2):155-160. ##Farfan-Ale JA, Lorono-Pino MA, Garcia-Rejon JE, Hovav E, Powers AM, Lin M, et al. Detection of RNA from a Novel West Nile-like virus and high prevalence of an insect-specific flavivirus in mosquitoes in the Yucatan Peninsula of Mexico. Am J Trop Med Hyg 2009;80(1):85-95. ##Beasley DW, Whiteman MC, Zhang S, Huang CY, Schneider BS, Smith DR, et al. Envelope protein glycosylation status influences mouse neuroinvasion phenotype of genetic lineage 1 West Nile virus strains. J Virol 2005;79(13):8339-8347. ##