How Does Iranian's Legal System Protect Human Vulnerability and Personal Integrity in Medical Research? 2 2 The astonishing advance of medical science in recent decades has had endless advantages for humans, including improved level of health, prevention of disease and advances in treatment. These advances depend to a great extent on conducting continuous research. However, besides its enormous advantages, the sole interest of medical science undermines the principles of respect for human vulnerability and personal integrity, in both positive and negative approaches. The positive approach refers to the people who participate in research and practice, while the negative approach refers to people who are deprived of research and practice. The authors of this work, based on legal or moral grounds try to analyse the tension between the principle of respect for human vulnerability and personal integrity and the interest of medical science. Undoubtedly, in applying scientific knowledge and medical practice human vulnerability should be taken into account. In this regard, especially vulnerable individuals and groups should be protected and the personal integrity of such individuals respected. In the light of the merits of Islamic law, this paper is designed to examine the significance of the principles of human vulnerability and personal integrity in medical research by studying the international documents as formalised by UNESCO in order to explore the place of these principles in the Iranian legal system. 51 60 Mohammad Taghi Karoubi University of Science and Culture, ACECR University of Science and Culture, ACECR Iran Mohammad Mehdi Akhondi Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran BioethicsIntegrityMedical ResearchVulnerability 60.pdf Coleman CH. Vulnerability as a regulatory category in human subject research. J Law, Med Ethics 2009;37(1):12-18. ##Ruof MC. Vulnerability, vulnerable populations and policy. Kennedy Inst Ethics J 2004;14(4):411-425. ##Murray TH, Mehlman MJ. Encyclopedia of Ethical, Legal, and Policy Issues in Biotechnology. New York: Wiley Interscience; 2000.##The Universal Declaration on Bioethics and Human Rights in 2005. ##Preliminary Draft Report of International Bioethics Committee on the Principle of Respect for Human Vulnerability and Personal Integrity, UNESCO, IBC., Paris, 2010.##Frewer S, Schmidt A. History and Theory of Human Experimentation: The Declaration of Helsinki and Modern Medical Ethics. Stuttgart: Franz Steiner Verlag; 2007.##Preliminary Draft Report on the Principle of Respect for Human Vulnerability and Personal Integrity, UNESCO, IBC., Mexico City, 2009.##Hurst S. Vulnerability in research and health care, describing the elephant in the room? Bioethics 2008;22(4):191-202. ##Roelcke V, Maio G (eds). Twentieth Century Ethics of Human Subjects Research. Historical Perspectives on Values, Practices, and Regulations. Stuttgart: Franz Steiner Verlag; 2004,162. ##Macklin R. Bioethics, vulnerability, and protection. Bioethics 2003;17(5-6):472-486.##Esposito JL. Islam and Politics. 3rd ed. New York: Syracuse University Press; 1991,84-86.##Fox K. Hotep's story: Exploring the wounds of health vulnerability in the US. Theor Med Bioethics 2002;23:471-497.##Hujjati Kirman MJ. Survey of the Similarities and Differences of Human Rights in Islam and the West. In: Islamic View on Human Rights. London: International Publisher; 2001,102.##Javadi Amuli A. Sources of Human Rights in Islam. In: Salimi H. Islamic View on Human Rights. London: Al-Hoda International Publisher; 2001,1-10. ##Bielefeldt H. Muslim voices in the human rights debate 17. Hum Rts Q 1995;587:601-606.##Soroush Mahalati, M. Roykard Keramat Mehvar be Fegh Islami, Moaseseseh Nashr va Asar Emam Khomini, 1386. (Persian).##Jafari MT. A Comparative Study of the Two Systems of Universal Human Rights. London: Al-Hoda International Publisher; 1999.##Hassan R. An Islamic Perspective. In: Becker J. Women Religion and Sexuality, Studies in the Impact of Religious Teachings on Women. Geneva: WLC Publications: 1990,93-128.##Saeed MA. Islamic Concept of Human Rights. In: Haider SM. Islamic Concept of Human Rights. Lahore: The Book House; 1978.##Mausumi SH. Islamic Concept of Human Rights. In: Haider SM. Islamic Concept of Human Rights. Lahore: The Book House; 1978.##Rahman S. Islamic Law: Its Scope and Equity. 1970.##Haider SM. Islamic Concept of Human Rights. Lahore: The Book House; 1978.##The Cairo Declaration on Human Rights in Islam in 1990. ##Mallat C. The Renewal of Islamic Law. Cambridge: Cambridge University Press; 1993.##Schirazi A. The Constitution of Iran, Politics and the State in Islamic Republic. London: I.B. Tauris; 1998.##Iran’s Ministry of Health and Medical Education. Protection Code of Human Subject in Medical Research. (Persian).##

miR-155 Down Regulation by LNA Inhibitor can Reduce Cell Growth and Proliferation in PC12 Cell Line 2 2 MicroRNAs (miRNAs) are a class of small non coding regulatory RNAs that have key functions in multiple cell processes. Deregulation of these tiny miRNAs are involved in various human diseases. MiR-155 is one of the multifunctional miRNA that its over-expression has been found to be associated with different kinds of cancer such as leukemia, breast and colon cancers. It is thought that deregulation and over-expression of this microRNA may be associated with PC12 cell proliferation. So, the aim of this study was to investigate the role of miR-155 expression on PC12 cell growth. For this reason, PC12 cells were cultured and transfected by 3 different concentration (25, 50 and 75 nmol) of either LNA anti-miR-155 or scramble antisense in 24-well plate. Then, total RNA was extracted from transfected cells. miRNA cDNAs were synthesized from isolated total RNA. In the second step, miR-155 expression level was analyzed using the quantitative real-time polymerase chain reaction (QRT-PCR). MTT test was performed to evaluate cell viability. In the next step, apoptosis assay was assessed to investigate anti miR-155 effect on PC12 cells death. Obtained results were analyzed with t-test. MTT test revealed that cell viability of transfected cells with 75 nM of anti-miR- 155 to be reduced by half of the control and scramble groups (0.5 vs. 0.97 and 0.94). Our data suggest that miR-155 over-expression is associated with PC12 cell growth. So, miR-155 down regulation by anti-miR-155 could open up new ways to restrain brain tumor growth, as anti-miR-155 causes PC12 cells to repress. 61 66 Fatemeh Kouhkan Department of Genetics, Faculty of Basic Sciences, Tarbiat Modares University Department of Genetics, Faculty of Basic Sciences, Tarbiat Modares University Iran Shaban Alizadeh Department of Hematology, Allied Medical School, Tehran University of Medical Sciences Department of Hematology, Allied Medical School, Tehran University of Medical Sciences Iran Saeid Kaviani Department of Hematology, School of Medicine, Tarbiat Modares University Department of Hematology, School of Medicine, Tarbiat Modares University Iran Masoud Soleimani Department of Hematology, School of Medicine, Tarbiat Modares University Department of Hematology, School of Medicine, Tarbiat Modares University Iran Ali Akbar Pourfathollah Immunology Department, School of Medicine, Tarbiat Modares University Immunology Department, School of Medicine, Tarbiat Modares University Iran Naser Amirizadeh Research Center of Iranian Blood Transfusion Organization (IBTO) Research Center of Iranian Blood Transfusion Organization (IBTO) Iran Saeid Abroun Department of Hematology, School of Medicine, Tarbiat Modares University Department of Hematology, School of Medicine, Tarbiat Modares University Iran Mehrdad Noruzinia Department of Hematology, School of Medicine, Tarbiat Modares University Department of Hematology, School of Medicine, Tarbiat Modares University Iran Shahin Mohamadi Department of Hematology, Allied Medical School, Tehran University of Medical Sciences Department of Hematology, Allied Medical School, Tehran University of Medical Sciences Iran ApoptosismiR-155MTT testPC12 cells 61.pdf Abba M, Allgayer H. MicroRNAs as regulatory molecules in cancer: a focus on models defining miRNA functions. Drug Discov Today Dis Models 2009;6(1):13-19.##Filipowicz W, Bhattacharyya SN, Sonenberg N. Mechanisms of post-transcriptional regulation by microRNAs: are the answers in sight? Nat Rev Genet 2008;9:102-114.##He L, Hannon GJ. MicroRNAs: small RNAs with a big role in gene regulation. Nat Rev Genet 2004;5:522-531.##Miska EA. How microRNAs control cell division, differentiation and death. Curr Opin Genet Dev 2005;15(5):563-568.##Nelson P, Kiriakidou M, Sharma A, Maniataki E, Mourelatos Z. The microRNA world: small is mighty. Trends Biochem Sci 2003;28(10):534-540.##Calin GA, Croce CM. (2006) MicroRNA signatures in human cancers. Nat Rev Cancer 2006:6:857-866.##Esquela-Kerscher A, Slack FJ. Oncomirs-microRNAs with a role in cancer. Nat Rev Cancer 2006;6:259-269. ##Garofalo M, Condorelli GL, Croce C, Condorelli G. MicroRNAs as regulators of death receptors signaling. Cell Death Differ 2010;17:200-208.##Lu J, Getz G, Miska EA, Alvarez-Saavedra E, Lamb J, Peck D, et al. MicroRNA expression profiles classify human cancers. Nature 2005;435: 834-838.##McManus MT. MicroRNAs and cancer. Semin Cancer Biol 2003;13(4):253-258.##Shenouda K, Alahari SK. MicroRNA function in cancer: oncogene or a tumor suppressor? Cancer Metastasis Rev 2009;28:369-378.##Jiang S, Zhang HW, Lu MH, He XH, Li Y, Gu H, et al. MicroRNA-155 functions as an oncomiR in breast cancer by targeting the suppressor of cytokine signaling 1 gene. Cancer Res 2010;70:3119-3125.##Dorsett Y, Tuschl T. siRNAs: applications in functional genomics and potential as therapeutics. Nat Rev Drug Discovery 2004;3:318-329.##Esau CC, Monia BP. Therapeutic potential for microRNAs. Adv Drug Deliv Rev 2007;59(2-3):101-114.##Mei J, Gao Y, Zhang L, Cai X, Qian H, Huang H, et al. VEGF-siRNA silencing induces apoptosis, inhibits proliferation and suppresses vasculogenic mimicry in osteosarcoma in vitro. Exp Oncol 2008;30(1):29-34.##Huang KH, Wu Y, Chen YT, Deng H, Lian GD, Shuai XT. PEI-PEG as a siRNA genetic vector demonstrating interference in the expression of CD44v6 protein in gastric cancer cells. Clin Oncol Cancer Res 2010;7(3):187-192.##Cheng Q, Wang WL, Yan W, Li QL, Wang L, Wang WY. Knockdown of survivin expression by siRNA induces apoptosis of hepatocellular carcinoma cell LINE SMMC-7721. World J Gastroenterol 2005;11(5):756-759##Patutina O, Mironova N, Popova N, Kaledin V, Nikolin V, Zenkova M. The siRNA targeted to mdr1b and mdr1a mRNAs in vivo sensitizes murine lymphosarcoma to chemotherapy. BMC Cancer 2010;10(1):204-206.##Zhang M, Zhou Y, Xie C, Zhou F, Chen Y, Han G, et al. STAT6 specific shRNA inhibits proliferation and induces apoptosis in colon cancer HT-29 cells. Cancer Lett 2006;243(1):38-46.##Pillai RS. MicroRNA function: multiple mechanisms for a tiny RNA? RNA 2005;11(12):1753-1761.##Singh SK, Pal Bhadra M, Girschick HJ, Bhadra, U. MicroRNAs-micro in size but macro in function. FEBS 2008: 275(20):4929-4944.##Xia PG. Great potential of microRNA in cancer stem cell. J Cancer Mol 2008;4(3):79-89.##Calin GA, Dumitru CD, Shimizu M, Bichi R, Rai K. Frequent deletions and down-regulation of microRNA genes miR-15 and miR-16 at 13q14 in chronic lymphocytic leukemia. Proc Natl Acad Sci U S A 2002;99:15524-15529.##Cimmino A, Calin GA, Fabbri M, Iorio MV, Ferracin M, Shimizu M, et al. miR-15 and miR-16 induce apoptosis by targeting BCL2. Proc Natl Acad Sci U S A 2005;102(39):13944-13949.##Faraoni I, Romana F, Cardone J, Bonmassar E. miR-155 gene: A typical multifunctional microRNA. Biochimica et Biophysica Acta (BBA) 2009;1792(6):497-505.##Kong W, He L, Coppola M, Guo J, Esposito NN, Coppola D, et al . MicroRNA-155 regulates cell survival, growth and chemosensitivity by targeting FOXO3a in breast cancer. J BioloChem 2010;285: 17869-17879.##Metzler M, Wilda M, Busch K, Viehmann S, Borkhardt A. High expression of precursor microRNA-155/BIC RNA in children with Burkitt lymphoma. Genes Chromosomes Cancer 2004;39(2):167-169.##Rai D, Karanti S, Jung D, Dahia PL, Aguiar RC. Coordinated expression of microRNA-155 and predicted target genes in diffuse large B-cell lymphoma. Cancer Genet 2008;181(1):8-15.##Rossi L, Bonmassar E, Faraoni I. Modification of miR gene expression pattern in human colon cancer cells following exposure to 5-fluorouracil in vitro. Pharmacol Res 2007;56(3):248-253.##Connell RM, Taganov KD, Boldin MP, Cheng G, Baltimore D. MicroRNA-155 is induced during the macrophage inflammatory response. Proc Natl Acad Sci U S A 2007;104(5):1604-1609.##Gironella M, Seux M, Xie MJ, Cano C, Tomasini R, Gommeaux J, et al. Tumor protein 53-induced nuclear protein 1 expression is repressed by miR-155, and its restoration inhibits pancreatic tumor development. Proc Natl Acad Sci U S A 2007;104(41):16170-16175.##Rodriguez A, Vigorito E, Clare S, Warren MV, Couttet P. Requirement of bic/microRNA-155 for normal immune function. Science 2007;316(5824):608-611.##

Editorial 2 2 Humans have used animals for nourishments, body parts to help survival, in sports, in fields, in experiments and also have domesticated them as companions for a long time. Although the animal rights groups have been raising the issue of animal cruelty for the past few decades, however, animals are still exploited in the industrial processing. Billions of chickens, turkeys, sheep, cows, pigs, geese, and other animals are maltreated everyday around the world in order to prepare them to feed humans. A vast literature exists on the issue of animal suffering and the cruelties humans exert on them, and its ethical implications have been discussed. However, there are conflicting viewpoints on animal experimentations and particularly genetic manipulations of animals in societies where biotechnology sector has reached a level of potentiality to produce such animals. For example, is it ethical to engineer hundreds of transgenic pigs and keep them in standby positions to provide heart valves and whole heart transplants? Or even engineer a pig to grow a heart using certain genes from the intended heart patients? Is it ethical to use animals entirely as a means to save a human? How is it that the animal can be eaten entirely for food purposes, but may be questioned ethically when the animal is to be helping a human to survive? In recent years, the biotechnology community has found the capacity to make any kind of transgenic animals they wish including: animals which glow green, become diabetic or oncogenic in a given span of time, serve as bioreactors or bodies that can produce drug molecules. The question is where to draw the line with regard to ethics in using biotechnology to manipulate animals in serving the needs of humans? Obviously, the lines are not quite clear and the standards may differ in different countries and cultures. But, what is clear is that as more societies find the capacity to create animals whose bodies are pharmaceutical manufacturing plants, whose organs provide transplants, and whose bodies are genetically altered to provide experimental platforms; the ethical bodies in the government and non-government should scrutinize the motives and actions where biotechnologies are applied. As some argue, the best way to measure our humanity is not only in respecting the individual human beings but also in how we treat animals in our societies. 66 66 Ali M. Ardekani Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Editorial 162.pdf ####

Expression and Single-step Purification of GRA8 Antigen of Toxoplasma gondii in Escherichia coli 2 2 Diagnosis of Toxoplasma gondii (T.gondii) infection is of great medical importance especially for pregnant women and immunosuppressed patients. Numerous studies have shown that the recombinant production of several tToxoplasma antigens, including dense granule antigens (GRAs) has a great potential as diagnostic reagents. Previous studies reported expression of amino terminal GRA8 protein in fusion with large tags. In the present study, we produced truncated GRA8 (TGRA8GRA8), excluded from the signal peptide and C-terminal transmembrane domain, with a short fusion tag in Escherichia coli. (E.coli). TGRA8GRA8 was purified using an optimized single-step Iimmobilized Mmetal ion Aaffinity Cchromatography (IMAC). The purity and yield of TGRA8GRA8 was highest at pH= 9.25. At this pH, 13.6 mg of TGRA8GRA8 was obtained with the purity of 97.97%. Immunogenicity of the protein was evaluated in Western blot analysis showing the serum sample from a rabbit immunized with TGRA8GRA8 recognized a single antigen of T. gondii tachyzoite at the expected molecular weight of native GRA8. To diagnosis acute tToxoplasma infection in pregnant women, an indirect immunoglobulin M (IgM) enzyme-linked immunosorbent assay (ELISA) was developed using TGRA8GRA8 resulting in a test specificity and sensitivity of 97.1% and 60.6%, respectively. These results demonstrated that immunogenic TGRA8GRA8 can be produced in fusion with a short tag and purified near to homogeneity using an optimized IMAC. TGRA8GRA8-IgM-ELISA was useful for detection of acute tToxoplasma infection. 67 78 Jalal Babaie Department of Parasitology, Pasteur Institute of Iran Iran Mandana Miri Department of Parasitology, Pasteur Institute of Iran Iran Ghazaleh Sadeghiani Department of Parasitology, Pasteur Institute of Iran Iran Mehrak Zare Department of Parasitology, Pasteur Institute of Iran Iran Ghader Khalili Department of Immunology, Pasteur Institute of Iran Iran Majid Golkar Department of Parasitology, Pasteur Institute of Iran Iran Enzyme-linked immunosorbent assayGene expressionGRA8 proteinImmunoglobulin MToxoplasma gondii 58.pdf Blader IJ, Saeij JP. Communication between Toxoplasma gondii and its host: impact on parasite growth, development, immune evasion, and virulence. APMIS 2009;117(5-6):458-476.##Montoya JG, Liesenfeld O. Toxoplasmosis. Lancet 2004;363:1965-1976.##Kotresha D, Noordin R. Recombinant proteins in the diagnosis of toxoplasmosis. APMIS 2010;118(8):529-542.##Sukthana Y. Toxoplasmosis: beyond animals to humans. Trends Parasitol 2006;22:137-142.##Wallon M, Kodjikian L, Binquet C, Garweg J, Fleury J, Quantin C, et al. Long-term ocular prognosis in 327 children with congenital toxoplasmosis. Pediatrics 2004;113(6):1567-1572.##Tenter AM, Heckeroth AR, Weiss LM. Toxoplasma gondii: from animals to humans. Int J Parasitol 2000;30(12-13):1217-1258.##Gatkowska J, Hiszczynska-Sawicka E, Kur J, Holec L, Dlugonska H. Toxoplasma gondii: an evaluation of diagnostic value of recombinant antigens in a murine model. Exp Parasitol 2006;114(3):220-227.##Beghetto E, Spadoni A, Bruno L, Buffolano W, Gargano N. Chimeric antigens of Toxoplasma gondii: toward standardization of toxoplasmosis serodiagnosis using recombinant products. J Clin. Microbiol 2006;44(6):2133-2140.##Montoya JG. Laboratory diagnosis of Toxoplasma gondii infection and toxoplasmosis. J Infect Dis 2002;185(Suppl 1):S73-S82.##Remington JS, Thulliez P, Montoya JG. Recent developments for diagnosis of toxoplasmosis. J Clin Microbiol 2004;42(3):941-945.##Hughes HP, Van Knapen F. Characterization of a secretory antigen from Toxoplasma gondii and its role in circulating antigen production. Int J Parasitol 1982;12(5):433-437.##Golkar M, Rafati S, Abdel-Latif MS, Brenier-Pinchart MP, Fricker-Hidalgo H, Sima BK, et al. The dense granule protein GRA2, a new marker for the serodiagnosis of acute Toxoplasma infection: comparison of sera collected in both France and Iran from pregnant women. Diagn Microbiol Infect Dis 2007;58(4):419-426.##Altcheh J, Diaz NS, Pepe CM, Martin V, Nigro M, Freilija H, et al. Kinetic analysis of the humoral immune response against 3 Toxoplasma gondii-recombinant proteins in infants with suspected congenital toxoplasmosis. Diagn Microbiol Infect Dis 2006;56(2):161-165.##Nigro M, Gutierrez A, Hoffer AM, Clemente M, Kaufer F, Carral L, et al. Evaluation of Toxoplasma gondii recombinant proteins for the diagnosis of recently acquired toxoplasmosis by an immunoglobulin G analysis. Diagn Microbiol Infect Dis 2003;47(4):609-613.##Li S, Maine G, Suzuki Y, Araujo FG, Galvan G, Remington JS, et al. Serodiagnosis of recently acquired Toxoplasma gondii infection with a recombinant antigen. J Clin Microbiol 2000;38(1):179-184.##Hiszczyjska-Sawicka E, Kur J, Pietkiewicz H, Holec L, Gasior A, Myjak P. Efficient production of the Toxoplasma gondii GRA6, p35 and SAG2 recombinant antigens and their applications in the serodiagnosis of toxoplasmosis. Acta Parasitol 2005;50(3):249–254.##Jacobs D, Vercammen M, Saman E. Evaluation of recombinant dense granule antigen 7 (GRA7) of Toxoplasma gondii for detection of immunoglobulin G antibodies and analysis of a major antigenic domain. Clin Diagn Lab Immunol 1999;6(1):24-29.##Pfrepper KI, Enders G, Gohl M, Krczal D, Hlobil H, Wassenberg D, et al. Seroreactivity to and avidity for recombinant antigens in toxoplasmosis. Clin Diagn Lab Immunol 2005;12(8):977-982.##Suzuki Y, Ramirez R, Press C, Li S, Parmley S, Thulliez P, et al. Detection of immunoglobulin M antibodies to P35 antigen of Toxoplasma gondii for serodiagnosis of recently acquired infection in pregnant women. J Clin Microbiol 2000;38(11):3967-3970.##Aubert D, Maine GT, Villena I, Hunt JC, Howard L, Sheu M, et al. Recombinant antigens to detect Toxoplasma gondii-specific immunoglobulin G and immunoglobulin M in human sera by enzyme immunoassay. J Clin Microbiol 2000;38(3):1144-1150.##Carey KL, Donahue CG, Ward GE. Identification and molecular characterization of GRA8, a novel, proline-rich, dense granule protein of Toxoplasma gondii. Mol Biochem Parasitol 2000;105(1):25-37.##Babaie J, Zare M, Sadeghiani Gh, Lorgard-Dezfuli M, Aghighi Z, Golkar M. Bacterial production of dense granule antigen GRA8 of Toxoplasma gondii. Iran Biomed J 2009;13(3):145-151##Li S, Galvan G, Araujo FG, Suzuki Y, Remington JS, Parmley S. Serodiagnosis of recently acquired Toxoplasma gondii infection using an enzyme-linked immunosorbent assay with a combination of recombinant antigens. Clin Diagn Lab Immunol 2000;7(5):781–787.##Porekar VG, Menart V. Perspective of immobilized metal affinity chromatography. J Biochem Biophys Met 2001;49(1-3):335-360.##Paunovic I, Schulin R, Nowack B. Evaluation of immobilized metal ion affinity chromatography for the fractionation of natural Cu complexing ligands. J Chromatogr 2005; 1100(2):176-184.##Sulkowski E. Purification of proteins by IMAC. Trends Biotechnol 1985;3(1):1-7.##Jiang W, Hearn MTW. Protein interaction with immobilized metal ion affinity ligands under high ionic strength conditions. Anal Biochem 1996;242(1):45-54.##Chaga GS. Twenty-five years of immobilized metal ion affinity chromatography: past, present and future. J Biochem Biophys Methods 2001;49(1-3):313-334.##Gaberc-Porekar V, Menart V. Perspectives of immobilized-metal affinity chromatography. J. Biochem. Biophys. Methods 2001; 49:335-360.##Charlton A, Zachariou M. Immobilized metal ion affinity chromatography of histidine-tagged fusion proteins. Methods Mol. Biol 2008;421:137-150.##Mercier C, Adjogble KD, Daubener W, Delauw MF. Dense granules: are they key organelles to help understand the parasitophorous vacuole of all Apicomplexa parasites?. Int J Parasitol 2005;35(8):829-849.##Golkar M, Shokrgozar MA, Rafati S, Sadaie MR, Assmar M. Construction, expression and preliminary immunological evaluation of a DNA plasmid encoding the GRA2 protein of Toxoplasma gondii. Iran Biomed J 2005;9(1):1-8.##Mercier C, Lecordier L, Darcy F, Deslee D, Murray A, Tourvieille B, et al. Molecular characterization of a dense granule antigen (Gra2) associated with the network of the parasitophorous vacuole in Toxoplasma gondii. Mol Biochem Parasitol 1993;58(1):71-82.##Parmley SF, Sgarlato GD, Remington JS. Genomic and corrected cDNA sequence of the P28 gene from Toxoplasma gondii. Mol Biochem Parasitol 1993;57:163-168.##Esposito D, Chatterjee DK. Enhancement of soluble protein expression through the use of fusion tags. Curr. Opin. Biotechnol 2006;17(4):353-358.##Tan YP, Ling TC, Tan WS, Yusoff Kh, Tey BT. Recovery of histidine-tagged nucleocapsid protein of Newcastle disease virus using immobilised metal affinity chromatography. Process Biochem 2006;41(4): 874-881.##Sickinger E, Andrieu FG, Jonas G, Schultess J, Stieler M, Smith D, et al. Performance characteristics of the new ARCHITECT Toxo IgG and Toxo IgG Avidity assays. Diagn Microbiol Infect Dis 2008;62(3):235-244##

Peptide-based Polyclonal Antibody Production against P110 Protein of Mycoplasma genitalium 2 2 Mycoplasma genitalium (M.genitalium) is a sexually transmitted pathogen. Detection of this microorganism in clinical specimens by culture is rather difficult and time consuming. The aim of this study was to produce polyclonal antibody against a synthetic peptide from P110 protein of M.genitalium in order to develop a diagnostic tool for detection of this microorganism in clinical specimens. A synthetic peptide from P110 protein was conjugated to Keyhole Limpet Hemocyanin )KLH( and used for immunization of a white New Zealand rabbit. The produced antibody was purified by affinity chromatography and its specific interaction with immunizing peptide was determined by ELISA. Immunoreactivity of the antibody was also tested by Western blotting in bacterial cell lysate prepared from M.genitalium G-37. To confirm its application as a diagnostic tool, indirect immunofluorescent staining method was performed on M.genitalium-infected PBMC using anti-P110 as the primary antibody. The results showed that produced antibody has excellent reactivity with immunizing peptide and also detected a single band of 110 kDa corresponding to P110 protein. M.genitalium-infected PBMC showed a bright fluorescent signal in IF staining. This antibody might be used as a tool in diagnostic applications. 79 86 Omid Zarei Department of Microbiology, School of Medicine, Tehran University of Medical SciencesMonoclonal Antibody Research Center, Avicenna Research Institute, ACECR Department of Microbiology, School of Medicine, Tehran University of Medical SciencesMonoclonal Antibody Research Center, Avicenna Research Institute, ACECR IranIran Gholam Reza Irajian Department of Microbiology, School of Medicine, Tehran University of Medical Sciences Department of Microbiology, School of Medicine, Tehran University of Medical Sciences Iran Amir-Hassan Zarnani Nanobiotechnology Research Center, Avicenna Research Institute, ACECRImmunology Research Center, School of Medicine, Tehran University of Medical Sciences Nanobiotechnology Research Center, Avicenna Research Institute, ACECRImmunology Research Center, School of Medicine, Tehran University of Medical Sciences IranIran Leili Chamani-Tabriz Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Shaghayegh Emami Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Monoclonal Antibody 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 Hodjattallah Rabbani Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Iran AntibodiesMycoplasma genitaliumPeptides 63.pdf Tully JG, Cole RM, Taylor-Robinson D, Rose DL. A newly discovered mycoplasma in the human urogenital tract. Lancet 1981;317(8233):1288-1291.##Tully JG, Taylor-Robinson D, Rose DL, Cole RM, Bove JM. Mycoplasma genitalium, a new species from the human urogenital tract. Int J Syst Bacteriol 1983;33:387-396. ##Jensen JS, Orsum R, Dohn B, Uldum S, Worm AM, Lind K. Mycoplasma genitalium: a cause of male urethritis? Genitourin Med 1993;69(4):265-269.##Totten PA, Schwartz MA, Sjostrom KE, Kenny GE, Handsfield HH, Weiss JB, et al. Association of Mycoplasma genitalium with nongonococcal urethritis in heterosexual men. J Infect Dis 2001;183(2):269-276.##Kreiger JN, Riley DE. Chronic prostatitis: Charlottesville to Seattle. J Urol 2004;172(6):2557-2560.##Mandar R, RaukasE, Turk S, Korrovits P, Punab M. Mycoplasmas in semen of chronic prostatitis patients. Scand J UrolNephrol 2005;39(6):479-482.##Namiki K, Goodison S, Porvasnik S, Allan RW, Iczkowski KA, Urbanek C, et al. Persistent exposure to Mycoplasma induces malignant transformation of human prostate cells. PloS One 2009;4(9):e6872.##Falk L, Fredlund H, Jensen JS. Signs and symptoms of urethritis and cervicitis among women with or without Mycoplasma genitalium or Chlamydia trachomatis infection. Sex Transm Infect 2005;81:73-78.##Taylor-Robinson D. Mycoplasma genitalium -an up-date. Int J STD AIDS 2002;13(3):145-151.##Cohen CR, Mugo NR, Astete SG, Odondo R, Manhart LE, Kiehlbauch JA, et al. Detection of Mycoplasma genitalium in women with laparoscopically diagnosed acute salpingitis. Sex Transm Infect 2005;81:463-466.##Ma L, Jensen JS, Myers L, Burnett J, Welch M, Jia Q, et al. Mycoplasma genitalium: an efficient strategy to generate genetic variation from a minimal genome. Mol Microbiol 2007;66(1):220-236.##Iverson-Cabral SL, Astete SG, Cohen CR, Totten PA. mgpB and mgpC sequence diversity in Mycoplasma genitalium is generated by segmental reciprocal recombination with repetitive chromosomal sequences. Mol Microbiol 2007;66(1):55-73.##Ma L, Jensen JS, Mancuso M, Hamasuna R, Jia Q, McGowin CL, et al. Genetic variation in the complete MgPa operon and its repetitive chromosomal elements in clinical strains of Mycoplasma genitalium. PLoS One 2010;5(12):e15660.##Hadavi R, Zarnani AH, Ahmadvand N, Mahmoudi AR, Bayat AA, Mahmoudian J, et al. Production of monoclonal antibody against human nestin. Avicenna J Med Biotech 2010;2(2):69-77.##Litvinov S. New adjuvant for accelerated and enhanced antibody response. Nature Methods 2009;6:10-11.##Tully JG, Rose DL, Whitcomb RF, Wenzel RP. Enhanced isolation of Mycoplasma pneumoniae from throat washings with a newly-modified culture medium. J Infect Dis 1979;139(4):478-482.##Ferrante A, Thong YH. 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Nephroprotective and Nitric oxide Scavenging Activity of Tubers of Momordica tuberosa in Rats 2 2 Hydroalcoholic extract (70% ethanol extract) of tubers of Momordica tuberosa Cogn. (Cucurbitaceae) was subjected to preliminary phytochemical screening by qualitative tests. Nitric oxide scavenging activity was performed by Griess reagent method. And nephroprotective activity was assessed in gentamicin, cisplatin and paracetamol induced renal damage in wistar rats (150-200 g) by standard methods. The protective property of 70% ethanol extract was assessed by measuring the levels of body weight, blood urea, serum creatinine, tissue glutathione and lipid peroxidation in administered doses. The extract exhibited free radical scavenging activity in dose dependant manner. And 100 g/ml dose produced significantly higher scavenging activity than standard sodium metabisulphate at 25 g/ml. 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Antidiarrheal, Antioxidant and Antimicrobial Activities of the Musa Sapientum Seed 2 2 Musa sapientum (M.sapientum) commonly known as ‘banana’ is widely used in Bangladeshi folk medicine for the treatment of various ailments including diarrhea. Hence, the present study was designed to investigate antidiarrheal, antioxidant and antibacterial potential of the methanolic extract of M.sapientum seed (MMSS). The extract was studied for antidiarrheal property using castor oil and magnesium sulfate induced diarrheal model and charcoal induced gastrointestinal motility test in mice. Total phenolic and flavonoids content, total antioxidant activity, scavenging of 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical, as well as nitric oxide (NO) and assessment of reducing power were used to evaluate antioxidant potential of MMSS. In addition, disc diffusion methods were used for antibacterial assay using various diarrheal induced bacterial strains. At the doses of 100 and 200 mg/kg body weight, the extract reduced the frequency and severity of diarrhea in test animals throughout the study period. At the same doses, the extracts significantly (p0.001) delayed the intestinal transit of charcoal meal in test animals as compared to the control. In DPPH and NO scavenging method, MMSS showed good antioxidant potentiality in a dose dependent manner with the IC50 value of 12.32±0.33 g/ml and 18.96±1.01 g/ml, respectively with a significant (p0.001) good reducing power. The extract also displayed strong antibacterial effect against when tested against Escherichia coli, Shigella dysenteriae, and Pseudomonas aeruginosa. Altogether, these results suggest that the MMSS could be used as a potential antidiarrheal agent along with its antioxidant and antibacterial potentiality. 95 105 Md. Sarowar Hossain Department of Pharmacy, Atish Dipankar University of Science & Technology Department of Pharmacy, Atish Dipankar University of Science & Technology Bangladesh Md. Badrul Alam Department of Pharmacy, Atish Dipankar University of Science & Technology Department of Pharmacy, Atish Dipankar University of Science & Technology Bangladesh Md. Asadujjaman Department of Pharmacy, Atish Dipankar University of Science & Technology Department of Pharmacy, Atish Dipankar University of Science & Technology Bangladesh Ronok Zahan Department of Pharmacy, Atish Dipankar University of Science & Technology Department of Pharmacy, Atish Dipankar University of Science & Technology Bangladesh M. Monirul Islam Department of Pharmacy, Atish Dipankar University of Science & Technology Department of Pharmacy, Atish Dipankar University of Science & Technology Bangladesh Mohammed Ehsanul Hoque Mazumder Faculty of Medicine, Cumberland Campus, University of Sydney Faculty of Medicine, Cumberland Campus, University of Sydney Australia Md. Ekramul Haque Department of Pharmacy, BRAC University Department of Pharmacy, BRAC University Bangladesh Antibacterial agentsDiarrheaFree radicalsMusa sapientum 59.pdf Guerrant RL, Van Gilder T, Steiner TS, Theilman MN, Slutsker L, Tauxe RV, et al. Practice guidelines for the management of infectious diarrhea. Clin Infec Dis 2001;32(3):331-351.##Suleiman MM, Dzenda T, Sani CA. Antidiarrhoeal activity of the methanol stem-bark extract of Annona senegalensis Pers. (Annonaceae). J Ethnopharmacol 2008;116(1):125-130. ##Kosek M, Bern C, Guerrant RL. The global burden of diarrheal disease, as estimated from studies published between 1992 and 2000. 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