Repositioning Drugs for Psychiatry 2 2 Drug development can be time-consuming and expensive. Recent estimates suggest that, on average, it takes 10 years and at least $1 billion to bring a drug to market. Since last decade, 30-40% of drugs or biologics that were approved or launched for the first time in the US were either drugs repositioned for new indications, reformulations or new combinations of existing drugs. This is the lifecycle business with repositioning as a major contributor, and it is rarely given much attention outside of its practitioners 1,2. In general, drug repurposing or drug repositioning alludes to the development of existing drugs or pro-drugs for new indications, not necessarily related to the original disease focus. These drugs have probably failed in late-stage clinical trials by lacking in efficacy or safety, or have problems associated with commercial strategies, patent expiration or geographic expansion. Repositioning existing drug substances for the treatment of different indications can significantly reduce the cost and time required for the development of new medicines. Therefore, drug repurposing brings forth the benefit of quickening patient access to innovative and effective treatment at lower risk and development cost for the industry 1,2. There are a number of different definitions of drug repurposing. All of them contain two key elements: Taking existing scientific or medical knowledge and technology that is "approved" for human use in one disease or condition; and Applying this knowledge and technology to another disease or condition. Aspirin, a drug that’s been in use in some form or other for many hundreds of years was originally employed, and indeed still is, as a mild pain-relieving analgesic. But it’s probably more commonly used today as an antiplatelet agent helping to prevent blood clotting that can occur in thromboembolic disease. Nervous system diseases represent a major health concern worldwide. Although important financial and professional investment, their etiology and pathophysiology still remain mostly elusive. Moreover, the clinical need of disease-modifying therapies is still unmet. In medicine in general and in psychiatry in particular, repositioning was the result of serendipitous but astute clinical observation of an unexpected benefit or expected or unexpected adverse effects 3. A number of old drugs have been reintroduced for psychiatric indications such as celecoxib for schizophrenia, tamoxifen for mania and scopolamine for depression 4-7. Drug repurposing has become a new business segment for the life science services industry. In conclusion, drug repurposing emerges as a new value proposition for the industry, patients and payers. 67 67 Shahin Akhondzadeh Editorial 20422.pdf Langedijk J, Mantel-Teeuwisse AK, Slijkerman DS, Schutjens MH. Drug repositioning and repurposing: terminology and definitions in literature. Drug Discov Today 2015;20(8):1027-1034.##Tiriveedhi V. Impact of precision medicine on drug repositioning and pricing: A too small to thrive crisis. J Pers Med 2018;8(4):36.##Hong J, Bang M. Anti-inflammatory strategies for schizophrenia: A review of evidence for therapeutic applications and drug repurposing. Clin Psychopharmacol Neurosci 2020;18(1):10-24.##Akhondzadeh S. The 5-HT hypothesis of schizophrenia. IDrugs 2001;4(3):295-300.##Abbasi SH, Behpournia H, Ghoreshi A, Salehi B, Raznahan M, Rezazadeh SA, et al. The effect of mirtazapine add on therapy to risperidone in the treatment of schizophrenia: a double-blind randomized placebo-controlled trial. Schizophr Res 2010;116(2-3):101-106.##Khajavi D, Farokhnia M, Modabbernia A, Ashrafi M, Abbasi SH, Tabrizi M, et al. Oral scopolamine augmentation in moderate to severe major depressive disorder: a randomized, double-blind, placebo-controlled study. J Clin Psychiatry 2012;73(11):1428-1433.##Abbasi SH, Hosseini F, Modabbernia A, Ashrafi M, Akhondzadeh S. Effect of celecoxib add-on treatment on symptoms and serum IL-6 concentrations in patients with major depressive disorder: randomized double-blind placebo-controlled study. J Affect Disord 2012;141(2-3):308-314.##

Bone Regeneration Using Bio-Nanocomposite Tissue Reinforced with Bioactive Nanoparticles for Femoral Defect Applications in Medicine 2 2 Background: In recent years, the method of constructing and evaluating the properties of polymer nanocomposite and bioactive ceramics in tissue engineering such as biocompatible scaffolds was studied by some researchers.  Methods: In this study, the bio-nanocomposite scaffolds of Chitosan (CS)–Hydroxya-patite (HA)–Wllastonite (WS), incorporated with 0, 10, 20 and 30 wt% of zirconium were produced using a freeze-drying method. Also, the phase structure and morphology of scaffolds were investigated using X-ray Diffraction (XRD), Scanning Electron Microscopy (SEM) and Energy Dispersive Spectroscopy (EDS). By analyzing the SEM images, the porosity of the scaffolds was observed in the normal bone area of the body. In the next step, bioactivity and biodegradability tests of the scaffolds were carried out. Due to the presence of hydrophilic components and the high-water absorption capacity of these materials, the bio-nanocomposite scaffolds were able to absorb water properly. After that, the mechanical properties of the scaffolds were studied. Results: The mechanical test results showed that the preparation of reinforced bio-nanocomposites containing 10 wt% of zirconium presented better properties compared to incorporated bio-nanocomposites with different loadings of zirconium. Conclusion: According to MTT assay results, the prepared scaffolds did not have cytotoxicity at different concentrations of scaffold extracts. Consequently, the investigated scaffold can be beneficial in bone tissue engineering applications because of its similarity to natural bone structure and its proper porosity. 68 76 Mohammad Ali Maghsoudlou Department of Pharmacology and Toxicology, AJA University of Medical Sciences Department of Pharmacology and Toxicology, AJA University of Medical Sciences Iran Ehsan Nassireslami Saeed Saber-Samandari New Technologies Research Center, Amirkabir University of Technology New Technologies Research Center, Amirkabir University of Technology Iran Amirsalar Khandan New Technologies Research Center, Amirkabir University of Technology New Technologies Research Center, Amirkabir University of Technology Iran Bone regenerationChitosanTissue engineering Zirconium 20406.pdf Braddock M, Houston P, Campbell C, Ashcroft P. Born again bone: tissue engineering for bone repair. News Physiology Sci 2001;16:208-213.##Goldberg VM. Natural history of autografts and allografts. 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Synthesis and characterization of porous cytocompatible scaffolds from polyvinyl alcohol–chitosan. Bull Mater Sci 2019;42:35.##Mututuvari TM, Harkins AL, Tran CD. Facile synthesis, characterization, and antimicrobial activity of cellulose–chitosan–hydroxyapatite composite material: A potential material for bone tissue engineering. J Biomed Mater Res A 2013;101(11):3266-3277.##Hajinasab A, Saber-Samandari S, Ahmadi S, Alamara K. Preparation and characterization of a biocompatible magnetic scaffold for biomedical engineering. Mater Chem Phys 2018;204:378-387.##Sahmani S, Saber-Samandari S, Khandan A, Aghdam M. Nonlinear resonance investigation of nanoclay based bio-nanocomposite scaffolds with enhanced properties for bone substitute applications. J Alloys Compd 2019;773:636-653.##Tripathi A, Saravanan S, Pattnaik S, Moorthi A, Partridge NC, Selvamurugan N. Bio-composite scaffolds containing chitosan/nano-hydroxyapatite/nano-copper–zinc for bone tissue engineering. Int J Biol Macromol 2012;50(1):294-299.##Beladi F, Saber-Samandari S, Saber-Samandari S. Cellular compatibility of nanocomposite scaffolds based on hydroxyapatite entrapped in cellulose network for bone repair. Mater Sci Eng C Mater Biol Appl 2017;75:385-392.##Karamian E, Motamedi MRK, Khandan A, Soltani P, Maghsoudi S. An in vitro evaluation of novel NHA/zircon plasma coating on 316L stainless steel dental implant. Prog Natural Sci Mater Int 2014;24(2):150-156.##Kordjamshidi A, Saber-Samandari S, Nejad MG, Khandan A. Preparation of novel porous calcium silicate scaffold loaded by celecoxib drug using freeze drying technique: Fabrication, characterization and simulation. Ceram Int 2019;45(11):14126-14135.##Sahmani S, Saber-Samandari S, Khandan A, Aghdam MM. Influence of MgO nanoparticles on the mechanical properties of coated hydroxyapatite nanocomposite scaffolds produced via space holder technique: fabrication, characterization and simulation. J Mech Behav Biomed Mater 2019;95:76-88.##Yu CC, Chang JJ, Lee YH, Lin YC, Wu MH, Yang MC, et al. Electrospun scaffolds composing of alginate, chitosan, collagen and hydroxyapatite for applying in bone tissue engineering. Mater Lett 2013;93:133-136.##Jin HH, Kim DH, Kim TW, Shin KK, Jung JS, Park HC, et al. In vivo evaluation of porous hydroxyapatite/chitosan–alginate composite scaffolds for bone tissue engineering. Int J Biol Macromol 2012;51(5):1079-1085.##Sahmani S, Shahali M, Khandan A, Saber-Samandari S, Aghdam M. Analytical and experimental analyses for mechanical and biological characteristics of novel nanoclay bio-nanocomposite scaffolds fabricated via space holder technique. Appl Clay Sci 2018;165:112-123.##Lee DB, Roberts M, Bluchel CG, Odell RA. Zirconium: biomedical and nephrological applications. ASAIO J 2010;56(6):550-556.##Ohtsuki C, Kokubo T, Yamamuro T. Mechanism of apatite formation on CaOSiO2P2O5 glasses in a simulated body fluid. 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Polyaniline Based Electrochemical Sensor for the Detection of Dengue Virus Infection 2 2 Background: Dengue burden is increasing day-by-day globally. A rapid, sensitive, cost-effective early diagnosis kit is the need of the hour. In this study, a label-free electrochemical immunosensor was proposed for dengue virus detection. A modified Polyaniline (PANI) coated Glassy Carbon (GC) electrode, immobilized with DENV NS1 antibody was used to detect the circulating DENV NS1 antigen in both spiked and infected sample. Methods: Cloning, purification and expression of DENV NS1 protein in Escherichia coli (E. coli) was performed and sensor design, PANI modification on GC electrode surface by electrochemical polymerization and immobilization of NS1 antibody on the modified electrode surface was done and finally the analytical performance of the electrochemical immunosensor was done using Cyclic Voltammetry (CV) and Electrochemical Impedance Spectroscopy (EIS). Results: CV and EIS were used to study and quantitate the circulating DENV antigen. The calibration curve showed wide linearity, good sensitivity (Slope=13.8% IpR/ml.ng-1) and distribution of data with a correlation coefficient (R) of 0.997. A lower Limit of Detection (LOD) was found to be 0.33 ng.ml-1 which encourages the applicability of the sensor. Conclusion: Thus, a PANI based new electrochemical immunosensor has been developed which has the potential to be further modified for the development of cost effective, point of care dengue diagnostic kit. 77 84 Reshmi Dutta Department of Biotechnology, Faculty of Bioengineering, SRM Institute of Science and Technology, Kattankulathur Department of Biotechnology, Faculty of Bioengineering, SRM Institute of Science and Technology, Kattankulathur India K Thangapandi Department of Biotechnology, National Institute of Technology, Yupia, Papum Pare Department of Biotechnology, National Institute of Technology, Yupia, Papum Pare India Sumantra Mondal Department of Biotechnology, Faculty of Bioengineering, SRM Institute of Science and Technology, Kattankulathur Department of Biotechnology, Faculty of Bioengineering, SRM Institute of Science and Technology, Kattankulathur India Amalesh Nanda Department of Biotechnology, National Institute of Technology, Yupia, Papum Pare Department of Biotechnology, National Institute of Technology, Yupia, Papum Pare India Shreyosi Bose Department of Biotechnology, Faculty of Bioengineering, SRM Institute of Science and Technology, Kattankulathur Department of Biotechnology, Faculty of Bioengineering, SRM Institute of Science and Technology, Kattankulathur India Shairee Sanyal Department of Biotechnology, Faculty of Bioengineering, SRM Institute of Science and Technology, Kattankulathur Department of Biotechnology, Faculty of Bioengineering, SRM Institute of Science and Technology, Kattankulathur India Saikat Kumar Jana Suvankar Ghorai Dengue virusDielectric spectroscopyElectrodesPolyanilineVoltammetry 10373.pdf Gubler DJ. 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Cysteine/Histidine-Dependent Amidohydrolase/Peptidase (CHAP)-Displayed Nano Phages: Antimicrobial Function against Methicillin-Resistant Staphylococcus aureus (MRSA) 2 2 Background: Emergence and prevalence of multi drug resistance strains such as Methicillin-Resistant Staphylococcus aureus (MRSA) call for new antibacterial option. Endolysins as a new option is suggested. The phage display technique is suggested for production of recombinant endolysins. The recombinant endolysins displayed nano phages specifically lysis bacteria, which penetrate to the depth of tissue and the effective dose is reduced. Methods: CHAPK gene was ligated in T7Select vector arms in T7Select10-3b cloning kit. To produce recombinant nano phages, ligation reaction was added directly to the packaging extract. Recombinant nano phages were amplified by Double Layer Agar assay (DLA). The recombinant nano phages were characterized using TEM. Size of recombinant nano phages was determined using DLS. The spot test was performed to confirm CHAPk -displayed on the surface of nano phages. The turbidimetry was used to investigate lytic activity of recombinant nano phages against MRSA ATCC No. 33591. Results: The results showed recombinant nano phages belonged to order Caudovirales and family Podoviridae with titer 2×107 PFU/ml. According to the results of DLS, size of recombinant nano phages was 71 nm. Formation inhibition zone confirmed the presence of CHAPk on the surface of nano phage phenotypically. The turbidimetry showed lytic activity recombinant nano phages against MRSA after 5 min. Conclusion: This study suggests that CHAPk -displayed nano phages can be effective in MRSA infections. 85 90 Golnar Rahimzadeh Pediatric Infectious Diseases Research Center, Mazandaran University of Medical Sciences Pediatric Infectious Diseases Research Center, Mazandaran University of Medical Sciences Iran Pooria Gill Nanomedicine Group, Immunogenetics Research Center, Mazandaran University of Medical Sciences Nanomedicine Group, Immunogenetics Research Center, Mazandaran University of Medical Sciences Iran Mohammad Sadegh Rezai BacteriophagesEndolysinMethicillin-resistant Staphylococcus aureus 20404.pdf Basak S, Singh P, Rajurkar M. Multidrug resistant and extensively drug resistant bacteria: A study. Journal of pathogens. J Pathog. 2016;2016:4065603.##Harsha P, Yogeshkumar V, Vyas BRM, Sumitra C. Emergence of methicillin-resistant staphylococcus aureus (MRSA) as a public-health threat and future directions of antibiotic therapy for MRSA infections. Antiinfect Agents. 2012;10(2):149-57.##Magiorakos AP, Srinivasan A, Carey R, Carmeli Y, Falagas M, Giske C, et al. Multidrug‐resistant, extensively drug‐resistant and pandrug‐resistant bacteria: an international expert proposal for interim standard definitions for acquired resistance. Clin Microbiol Infect. 2012;18(3):268-81.##Nobrega FL, Costa AR, Kluskens LD, Azeredo J. Revisiting phage therapy: new applications for old resources. Trends Microbiol 2015;23(4):185-191.##Rahimzadeh G, Gill P, Rezai MS. Characterization and lytic activity of methicillin-resistant Staphylococcus aureus (MRSA) phages isolated from NICU. Australas Med J 2016;9(6):169.##Rahimzadeh G, Gill P, Rezai MS. Characterization of methicillin-resistant Staphylococcus aureus (MRSA) phages from sewage at a tertiary pediatric hospital. Arch Pediatr Infect Dis 2016;5(1):e39615##Rahimzadeh G, Gill P, Rezai MS. Endolysins of bacteriophages as an anti-methicillin resistant Staphylococcus aureus infection in children: a narrative review. J Pediatr Rev 2018;6(1):36-43.##Rezai MS, G Rahimzadeh, P Gill. Fabrication of phage nanobioparticles against burn wound methicillin-resistant Staphylococcus aureus (MRSA) infections. WO 2019/ 162972 A1.##Rahimzadeh G, Saeedi M, Farshidi F, Rezai MS. Phage therapy in treatment of gram-negative bacterial infections: a systematic review. J Mazandaran Univ Med Sci 2018;28(165):203-212.##Borysowski J, Weber-Dąbrowska B, Górski A. Bacteriophage endolysins as a novel class of antibacterial agents. Exp Biol Med (Maywood) 2006;231(4):366-377.##Fischetti VA. Bacteriophage lysins as effective antibacterials. Curr Opin Microbiol 2008;11(5):393-400.##Fischetti VA. Bacteriophage endolysins: a novel anti-infective to control Gram-positive pathogens. Int J Med Microbiol 2010;300(6):357-362.##Daniel A, Euler C, Collin M, Chahales P, Gorelick KJ, Fischetti VA. 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Design of Anti-Angiogenic Peptidomimetics and Evaluation their Biological Activity by In Vitro Assays 2 2 Background: One of the important therapeutic approaches in cancer field is development of compounds which can block the initial tumor growth and the progression of tumor metastasis with no side effects. Thus, the recent study was carried out to design anti-VEGFR2-peptidomimetics as the most significant factor of angiogenesis process- and evaluate their biological activity by in vitro assays. Methods: We designed anti-VEGFR2 peptidomimetics with anti-angiogenic activity, including compound P (lactam derivative) and compound T (indole derivative) by using in silico methods. Then, the inhibitory activity on angiogenesis was evaluated by using angiogenesis specific assays such as Human Umbilical Vein Endothelial Cell (HUVEC) proliferation, tube formation in Matrigel, MTT and Real-Time PCR. IC50 values of the compounds were also determined by cytotoxicity plot in MTT assay. Results: Compounds P and T inhibited HUVEC cell proliferation and viability in a dose-dependent manner. The IC50 for compound T and compound P in HUVEC cell line were 113 and 115 μg/ml, respectively. Tube formation assay revealed that both compounds can inhibit angiogenesis effectively. The results of Real-Time PCR also showed these compounds are able to inhibit the expression of CD31 gene in HUVEC cell line. Conclusion: Our study suggested that compounds P and T may act as therapeutic molecules, or lead compounds for development of angiogenesis inhibitors in VEGF-related diseases. 91 98 Mona Ghadam Department of Medical Biotechnology, Biotechnology Research Center, Pasteur Institute of Iran Iran Soroush Sardari Mohammad Ali Shokrgozar Mahdiyeh Sadat Mahdavi Department of Medical Biotechnology, Biotechnology Research Center, Pasteur Institute of Iran Iran Angiogenesis inhibitorsDrug designPeptidomimeticsVascular endothelial growth factor receptor 20409.pdf Ferrara N. 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Identification of a peptide blocking vascular endothelial growth factor (VEGF)-mediated angiogenesis. EMBO J 2000;19(7):1525-1533.##Bainbridge JW, Jia H, Bagherzadeh A, Selwood D, Ali RR, Zachary I. A peptide encoded by exon 6 of VEGF (EG3306) inhibits VEGF-induced angiogenesis in vitro and ischaemic retinal neovascularisation in vivo. Biochem Biophys Res Commun 2003;302(4):793-799.##Mizejewski GJ. Peptides as receptor ligand drugs and their relationship to G-coupled signal transduction. Expert Opin Investig Drugs 2001;10(6):1063-1073.##Sillerud LO, Larson RS. Design and structure of peptide and peptidomimetic antagonists of protein-protein interaction. Curr Protein Pept Sci 2005;6(2):151-169.##Nestor JJ Jr. The medicinal chemistry of peptides. Curr Med Chem 2009;16(33):4399-4418.##Méndez-Samperio P. Peptidomimetics as a new generation of antimicrobial agents: current progress. Infect Drug Resist 2014;7:229-237.##Gokhale AS, Satyanarayanajois S. Peptides and peptidomimetics as immunomodulators. Immunotherapy 2014;6(6):755-774.##Kaumaya PT, Foy KC. Peptide vaccines and peptidomimetics targeting HER and VEGF proteins may offer a potentially new paradigm in cancer immunotherapy. Future Oncol 2012;8(8):961-987.##Bruno BJ, Miller GD, Lim CS. Basics and recent advances in peptide and protein drug delivery. Ther Deliv 2013;4(11):1443-1467.##Sulochana KN, Ge R. Developing antiangiogenic peptide drugs for angiogenesis-related diseases. Curr Pharm Des 2007;13(20):2074-2086.##Goede A, Michalsky E, Schmidt U, Preissner R. SuperMimic--fitting peptide mimetics into protein structures. BMC Bioinformatics 2006;7:11.##Divya PS, Jain K, Sobhia ME. From peptides to peptidomimetics: rational design of potential PKC-β II inhibitors. Med Chem Res 2013;22(2):625-634.##Kim JW, Kim TD, Hong BS, Kim OY, Yoon WH, Chae CB, et al. A serum-stable branched dimeric anti-VEGF peptide blocks tumor growth via anti-angiogenic activity. 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Methylation Analysis of P16, RASSF1A, RPRM, and RUNX3 in Circulating Cell-Free DNA for Detection of Gastric Cancer: A Validation Study 2 2 Background: Most of Gastric Cancer (GC) patients are diagnosed at an advanced stage with poor prognosis. Hypermethylations of several tumor suppressor genes in cell-free DNA of GC patients have been previously reported. In this study, an attempt was made to investigate the methylation status of P16, RASSF1A, RPRM, and RUNX3 and their potentials for early diagnosis of GC. Methods: Methylation status of the four tumor suppressor genes in 96 plasma samples from histopathologically confirmed gastric adenocarcinoma patients (Stage I-IV) and 88 healthy controls was determined using methylation-specific PCR method. Receiver operating characteristic curve analysis was performed and Area Under the Curve (AUC) was calculated. Two tailed p<0.05 were considered statistically significant. Results: Methylated P16, RASSF1A, RPRM, and RUNX3 were significantly higher in the GC patients (41.7, 33.3, 66.7, and 58.3%) compared to the controls (15.9, 0.0, 6.8, and 4.5%), respectively (p<0.001). Stratification of patients showed that RPRM (AUC: 0.70, Sensitivity: 0.47, Specificity: 0.93, and p<0.001) and RUNX3 (AUC: 0.77, Sensitivity: 0.59, Specificity: 0.95, and p<0.001) had the highest performances in detection of early-stage (I+II) GC. The combined methylation of RPRM and RUNX3 in detection of early-stage GC had a higher AUC of 0.88 (SE=0.042; 95% CI:0.793–0.957; p<0.001), higher sensitivity of 0.82 and reduced specificity of 0.89. Conclusion: Methylation analysis of RPRM and RUNX3 in circulating cell free-DNA of plasma could be suggested as a potential biomarker for detection of GC in early-stages. 99 106 Kioomars Saliminejad Hematology, Oncology and Stem Cell Transplantation Research Center, Shariati Hospital, Tehran University of Medical SciencesReproductive Biotechnology Research Center, Avicenna Research Institute (ACECR) Hematology, Oncology and Stem Cell Transplantation Research Center, Shariati Hospital, Tehran University of Medical Sciences IranIran Shahrzad Soleymani Fard Hematology, Oncology and Stem Cell Transplantation Research Center, Shariati Hospital, Tehran University of Medical Sciences Hematology, Oncology and Stem Cell Transplantation Research Center, Shariati Hospital, Tehran University of Medical Sciences Iran Hamid Reza Khorram Khorshid Genetics Research Center, University of Social Welfare and Rehabilitation Sciences, Tehran, Iran Genetics Research Center, University of Social Welfare and Rehabilitation Sciences, Tehran, Iran Iran Marjan Yaghmaei Hematology, Oncology and Stem Cell Transplantation Research Center, Shariati Hospital, Tehran University of Medical Sciences Hematology, Oncology and Stem Cell Transplantation Research Center, Shariati Hospital, Tehran University of Medical Sciences Iran Habibollah Mahmoodzadeh Department of Surgery, Cancer Institute, Imam Khomeini Hospital, Tehran, University of Medical Sciences Department of Surgery, Cancer Institute, Imam Khomeini Hospital, Tehran, University of Medical Sciences Iran Seyed Asadollah Mousavi Hematology, Oncology and Stem Cell Transplantation Research Center, Shariati Hospital, Tehran University of Medical Sciences Hematology, Oncology and Stem Cell Transplantation Research Center, Shariati Hospital, Tehran University of Medical Sciences Iran Seyed Hamidollah Ghaffari BiomarkersCell-free DNAGastric cancerDNA methylation 10370.pdf Ferlay J, Soerjomataram I, Dikshit R, Eser S, Mathers C, Rebelo M, et al. Cancer incidence and mortality worldwide: sources, methods and major patterns in GLOBOCAN 2012. Int J Cancer 2015;136(5):E359-386.##Uchôa Guimarães CT, Ferreira Martins NN, Cristina da Silva Oliveira K, Almeida CM, Pinheiro TM, Gigek CO, et al. Liquid biopsy provides new insights into gastric cancer. Oncotarget 2018;9(19):15144-15156.##Levy I, Gralnek IM. Complications of diagnostic colonoscopy, upper endoscopy, and enteroscopy. Best Pract Res Clin Gastroenterol 2016;30(5):705-718.##Gerlinger M, Rowan AJ, Horswell S, Math M, Larkin J, Endesfelder D, et al. Intratumor heterogeneity and branched evolution revealed by multiregion sequencing. N Engl J Med 2012;366(10):883-892.##Vedeld HM, Goel A, Lind GE. Epigenetic biomarkers in gastrointestinal cancers: The current state and clinical perspectives. Semin Cancer Biol 2018;51:36-49.##Toiyama Y, Okugawa Y, Goel A. DNA methylation and microRNA biomarkers for noninvasive detection of gastric and colorectal cancer. Biochem Biophys Res Commun 2014;455(1-2):43-57.##Padmanabhan N, Ushijima T, Tan P. How to stomach an epigenetic insult: the gastric cancer epigenome. Nat Rev Gastroenterol Hepatol 2017;14(8):467-478.##Hu W, Zheng W, Liu Q, Chu H, Chen S, Kim JJ, et al. Diagnostic accuracy of DNA methylation in detection of gastric cancer: a meta-analysis. Oncotarget 2017;8(68):113142-113152.##Issaq HJ, Waybright TJ, Veenstra TD. Cancer biomarker discovery: Opportunities and pitfalls in analytical methods. Electrophoresis 2011;32(9):967-975.##Pepe MS, Etzioni R, Feng Z, Potter JD, Thompson ML, Thornquist M, et al. Phases of biomarker development for early detection of cancer. J Natl Cancer Inst 2001;93(14):1054-1061.##Hu SL, Kong XY, Cheng ZD, Sun YB, Shen G, Xu WP, et al. Promoter methylation of p16, Runx3, DAPK and CHFR genes is frequent in gastric carcinoma. Tumori 2010;96(5):726-733.##do Nascimento Borges B, Burbano RM, Harada ML. Analysis of the methylation patterns of the p16 INK4A, p15 INK4B, and APC genes in gastric adenocarcinoma patients from a Brazilian population. Tumour Biol 2013;34(4):2127-2133.##Ye M, Xia B, Guo Q, Zhou F, Zhang X. Association of diminished expression of RASSF1A with promoter methylation in primary gastric cancer from patients of central China. BMC Cancer 2007;7:120.##Ooki A, Yamashita K, Yamaguchi K, Mondal A, Nishimiya H, Watanabe M. DNA damage-inducible gene, reprimo functions as a tumor suppressor and is suppressed by promoter methylation in gastric cancer. Mol Cancer Res 2013;11(11):1362-1374.##Sperka T, Wang J, Rudolph KL. DNA damage checkpoints in stem cells, ageing and cancer. Nat Rev Mol Cell Biol 2012;13(9):579-590.##Qu Y, Dang S, Hou P. Gene methylation in gastric cancer. Clin Chim Acta 2013;424:53-65.##Amigo JD, Opazo JC, Jorquera R, Wichmann IA, Garcia-Bloj BA, Alarcon MA, et al. The reprimo gene family: a novel gene lineage in gastric cancer with tumor suppressive properties. Int J Mol Sci 2018;19(7). E1862.##Chen F, Liu X, Bai J, Pei D, Zheng J. The emerging role of RUNX3 in cancer metastasis (Review). Oncol Rep 2016;35(3):1227-1236.##Balgkouranidou I, Matthaios D, Karayiannakis A, Bolanaki H, Michailidis P, Xenidis N, et al. Prognostic role of APC and RASSF1A promoter methylation status in Cell-free circulating DNA of operable gastric cancer patients. Mutat Res 2015;778:46-51.##Lin Z, Luo M, Chen X, He X, Qian Y, Lai S, et al. Combined detection of plasma ZIC1, HOXD10 and RUNX3 methylation is a promising strategy for early detection of gastric cancer and precancerous lesions. J Cancer 2017;8(6):1038-1044.##Liu L, Yang X. Implication of reprimo and hMLH1 gene methylation in early diagnosis of gastric carcinoma. Int J Clin Exp Pathol 2015;8(11):14977-14982.##Wang H, Zheng Y, Lai J, Luo Q, Ke H, Chen Q. Methylation-sensitive melt curve analysis of the reprimo Gene methylation in gastric cancer. PLoS One 2016;11(12):e0168635.##Wu YC, Lv P, Han J, Yu JL, Zhu X, Hong LL, et al. Enhanced serum methylated p16 DNAs is associated with the progression of gastric cancer. Int J Clin Exp Pathol 2014;7(4):1553-1562.##Abbaszadegan MR, Moaven O, Sima HR, Ghafarzadegan K, A'rabi A, Forghani MN, et al. p16 promoter hypermethylation: a useful serum marker for early detection of gastric cancer. World J Gastroenterol 2008;14(13):2055-2060.##Lee TL, Leung WK, Chan MW, Ng EK, Tong JH, Lo KW, et al. Detection of gene promoter hypermethylation in the tumor and serum of patients with gastric carcinoma. Clin Cancer Res 2002;8(6):1761-1766.##Pimson C, Ekalaksananan T, Pientong C, Promthet S, Putthanachote N, Suwanrungruang K, et al. Aberrant methylation of PCDH10 and RASSF1A genes in blood samples for non-invasive diagnosis and prognostic assessment of gastric cancer. 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Analysis of Glioblastoma Multiforme Tumor Metabolites Using Multivoxel Magnetic Resonance Spectroscopy 2 2 Background: Glioblastoma Multiforme (GBM) is the most common and deadly type of primary brain tumor in adults. Magnetic Resonance Spectroscopy (MRS) is a non-invasive imaging technique used to study metabolic changes in the brain tumors. Some metabolites such as Phosphocholine, Creatine, NAA/Cr, and Pcho/Cr have been proven to show a diagnostic role in GBM. The present study was conducted to analyze important metabolites using MRS multivoxel in GBM tumor. Methods: In this study, information was collected from 8 individuals diagnosed with GBM using Siemens multivoxel MRS with a magnetic field strength of 3 T. Data were obtained by Point-Resolved Spectroscopy (PRESS) protocol with TE=135 ms and TR=1570 ms. NAA, Pcho, Cr, Ala, Gln, Gly, Glu, Lac, NAAG, and Tau metabolites were extracted and evaluated statistically. Results: Given total number of normal voxels and total number of all voxels, levels of Cr, Glu, NAA, NAAG, and Gly/Tau ratio in healthy voxels were significantly higher than tumoral voxels (p=0.005, p=0.03, p<0.001, p<0.001 and p=0.041, respectively). In contrast, levels of Gly, Gln, Tau, Lac/Cr, Pcho/Cr, Pcho/NAA, Lac/NAA, and Gln/Glu ratios in tumoral voxels were significantly more than healthy voxels (p=0.001, p=0.037, p<0.001, p=0.010, p<0.001, p<0.001, and p=0.024, respectively). However, levels of Lac and Pcho had no significant difference in the two types of voxels. Conclusion: In summary, compared to patients with glioblastoma with 1H-MRS, the Pcho/Cr and Pcho/NAA ratios, and NAAG are the most important parameters to differentiate between tumoral and normal voxels. 107 115 Meysam Siyah Mansoory Ayob Faramarzi Department of Biomedical Engineering, Faculty of Medicine, Kermanshah University of Medical Sciences Department of Biomedical Engineering, Faculty of Medicine, Kermanshah University of Medical Sciences Iran Karim Khoshgard Pharmaceutical Sciences Research Center, Kermanshah University of Medical SciencesDepartment of Medical Physics, Faculty of Medicine, Kermanshah University of Medical Sciences Pharmaceutical Sciences Research Center, Kermanshah University of Medical SciencesDepartment of Medical Physics, Faculty of Medicine, Kermanshah University of Medical Sciences IranIran Hadi Mozafari Medical Biology Research Center, Kermanshah University of Medical Sciences Medical Biology Research Center, Kermanshah University of Medical Sciences Iran Glioblastoma multiformMagnetic resonance spectroscopyNeurochemical profilesVoxel 20407.pdf Tome-Garcia J, Tejero R, Nudelman G, Yong RL, Sebra R, Wang H, et al. 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Optimization of Fermentation Conditions to Enhance Cytotoxic Metabolites Production by Bacillus velezensis Strain RP137 from the Persian Gulf 2 2 Background: Isolation, introduction and producing bioactive compounds from bacteria, especially marine bacteria, is an attractive research area. One of the main challenges of using these metabolites as drug and their industrialization is the optimization of production conditions. Methods: In the present study, the response surface methodology was applied to optimize the production of a cytotoxic extract (C-137-R) by Bacillus velezensis (B. velezensis) strain RP137. Initially, among the three carbon and three nitrogen sources, rice starch and potassium nitrate were selected as the best, with cell toxicity equal to IC50=54.4 and 45.1 μg/ml in human lung and liver cancer cell lines, respectively (A549 and HepG2). In the next step, fractional factorial design was performed to survey effect of seven physical and chemical factors on the amount of production, and the most important factors including carbon and nitrogen sources with the positive effect and the sea salt with negative effect were determined. Finally, using the central composite design with 20 experiments, the best concentrations of rice starch and potassium nitrate (1.5%) and sea salt (1%) were obtained. Results: The average amount of dried extract produced in the optimum conditions was 131.1 mg/L and the best response was 71.45%, which is more than 28-fold better than the pre-optimized conditions. Conclusion: In general, it can be suggested that the use of modern statistical methods to optimize environmental conditions affecting the growth and metabolism of bacteria can be a highly valuable tool in industrializing the production of bioactive compounds. 116 123 Roya Pournejati Molecular Biotechnology Laboratory, Department of Biology, Faculty of Science, Shiraz University Molecular Biotechnology Laboratory, Department of Biology, Faculty of Science, Shiraz University Iran Hamid Reza Karbalaei-Heidari A549 cellsBacillusIndustrial developmentLiver neoplasms 20405.pdf Osbourn A. Secondary metabolic gene clusters: evolutionary toolkits for chemical innovation. Trends Genet 2010;26(10):449-457.##Tenconi E, Traxler M, Hoebreck C, van Wezel GP, Rigali S. Prodiginine production in Streptomyces coelicolor correlates temporally and spatially to programmed cell death. bioRxivorg 2018;240689.##Singh V, Haque S, Khare S, Tiwari AK, Katiyar D, Banerjee B, Kumari K, Tripathi C. Isolation and purification of antibacterial compound from Streptomyces levis collected from soil sample of north India. PLoS One 2018;13(7): e0200500.##Challinor VL, Bode HB. Bioactive natural products from novel microbial sources. Ann N Y Acad Sci 2015;1354:82-97.##Hoshino S, Zhang L, Awakawa T, Wakimoto T, Onaka H, Abe I. Arcyriaflavin E, a new cytotoxic indolocarbazole alkaloid isolated by combined-culture of mycolic acid-containing bacteria and streptomyces cinnamoneus NBRC 13823. J Antibiot (Tokyo) 2015;68(5):342-344.##Asolkar RN, Singh A, Jensen PR, Aalbersberg W, Carté BK, Feussner KD, et al. Marinocyanins, cytotoxic bromo-phenazinone meroterpenoids from a marine bacterium from the streptomycete clade MAR4. Tetrahedron 2017;73(16):2234-2241.##McCormick JR, Flärdh K. Signals and regulators that govern streptomyces development. FEMS Microbiol Rev 2012;36(1):206-231.##Na LI, Weiyan DU, Huang Z, Wei Z, Shoujiang W. Effect of imidazolium ionic liquids on the hydrolytic activity of lipase. Chinese J Cat 2013;34(4):769-780.##van Wezel GP, McDowall KJ. The regulation of the secondary metabolism of Streptomyces: new links and experimental advances. Nat Prod Rep 2011;28(7):1311-1333.##Ryu YG, Butler MJ, Chater KF, Lee KJ. Engineering of primary carbohydrate metabolism for increased production of actinorhodin in streptomyces coelicolor. Appl Environ Microbiol 2006;72(11):7132-7139.##Nguyen QT, Merlo ME, Medema MH, Jankevics A, Breitling R, Takano E. Metabolomics methods for the synthetic biology of secondary metabolism. FEBS Lett 2012;586(15):2177-2183.##Demain AL, Sanchez S. Microbial drug discovery: 80 years of progress. J Antibiot (Tokyo) 2009;62(1):5-16.##Rokem JS, Lantz AE, Nielsen J. Systems biology of antibiotic production by microorganisms. Nat Prod Rep 2007;24(6):1262-1287.##Scherlach K, Hertweck C. Triggering cryptic natural product biosynthesis in microorganisms. Org Biomol Chem 2009;7(9):1753-1760.##Feng J, Zhang W, Han L, Zhang X. Statistical optimization of medium components to improve the antibiotic activity of Streptomyces sp. 19G-317. Afr J Agric Res 2011;6(19):4424-4431.##Grahovac J, Grahovac M, Dodić J, Bajić B, Balaž J. Optimization of cultivation medium for enhanced production of antifungal metabolites by Streptomyces hygroscopicus. Crop Prot 2014;65:143-152.##Bankar SB, Singhal RS. Optimization of poly-epsilon-lysine production by streptomyces noursei NRRL 5126. Bioresour Technol 2010;101(21):8370-8375.##Purama RK, Goyal A. Screening and optimization of nutritional factors for higher dextransucrase production by Leuconostocmesenteroides NRRL B-640 using statistical approach. Bioresour Technol 2008;99(15):7108-7114.##Thi Nguyen HY, Tran GB. Optimization of fermentation conditions and media for production of glucose isomerase from Bacillus megaterium using response surface methodology. Scientifica (Cairo) 2018;2018:6842843.##Beigi L, Karbalaei-Heidari HR, Kharrati-Kopaei M. Optimization of an extracellular zinc-metalloprotease (SVP2) expression in Escherichia coli BL21 (DE3) using response surface methodology. Protein Expr Purif 2012;84(1):161-166.##Qiu JJ, Chen W, Ding M, Zhang ML, Zhao FK. Optimization of penicillin G acylase production by recombinat bacilllus subtilis via response surface analysis. J Zhejiang Sci Tech Univ 2012;29(6):868-873.##Pournejati R, Gust R, Karbalaei-Heidari HR. An aminoglycoside antibacterial substance, S-137-R, produced by newly isolated Bacillus velezensis strain RP137 from the Persian Gulf. Curr Microbiol 2019;76(9):1028-1037.##Van Meerloo J, Kaspers GJ, Cloos J. Cell sensitivity assays: the MTT assay. In: Cancer Cell Culture. Humana Press; 2011. p.237-245##Calcagno AM, Ambudkar SV. Molecular mechanisms of drug resistance in single-step and multi-step drug-selected cancer cells. Methods Mol Biol 2010;596:77-93.##Yu J, Liu Q, Liu Q, Liu X, Sun Q, Yan J, et al. Effect of liquid culture requirements on antifungal antibiotic production by Streptomyces rimosus MY02. Bioresour Technol 2008;99(6):2087-2091.##Yuan LL, Li YQ, Wang Y, Zhang XH, Xu YQ. Optimization of critical medium components using response surface methodology for phenazine-1-carboxylic acid production by Pseudomonas sp. M-18Q. J Biosci Bioeng 2008;105(3):232-237.##Demain AL. in Ciba Foundation Symposium 171‐Secondary Metabolites: their Function and Evolution: Secondary Metabolites: Their Function and Evolution: Ciba Foundation Symposium 171. 3-23 (Wiley Online Library).##Satoh A, Ogawa H, Satomura Y. Regulation of N-acetyl kanamycin amidohydrolase in the idiophase in kanamycin fermentation. Agr Biol Chem 1976;40(1):191-196.##Ruiz B, Chávez A, Forero A, García-Huante Y, Romero A, Sánchez M, et al. Production of microbial secondary metabolites: regulation by the carbon source. Crit Rev Microbiol 2010;36(2):146-167.##Jonsbu E, McIntyre M, Nielsen J. The influence of carbon sources and morphology on nystatin production by Streptomyces noursei. J Biotechnol 2002;95(2):133-144.##Rastegari B, Karbalaei-Heidari HR. Sulfate as a pivotal factor in regulation of Serratia sp. strain S2B pigment biosynthesis. Res Mcrobiol 2016;167(8):638-646.##Ahsan T, Chen J, Wu Y, Irfan M. Application of response surface methodology for optimization of medium components for the production of secondary metabolites by Streptomyces diastatochromogenes KX852460. AMB Express 2017;7(1):96.##Voelker F, Altaba S. Nitrogen source governs the patterns of growth and pristinamycin production in ‘streptomyces pristinaespiralis’. Microbiology 2001;147(Pt 9):2447-2459.##Kavitha G, Kurinjimalar C, Sivakumar K, Kaarthik M, Aravind R, Palani P, et al. Optimization of polyhydroxybutyrate production utilizing waste water as nutrient source by botryococcus braunii Kütz using response surface methodology. Int J Biol Macromol 2016;93(Pt A):534-542.##Kong Y, Zou P, Miao L, Qi J, Song L, Zhu L, et al. Medium optimization for the production of anti-cyanobacterial substances by streptomyces sp. HJC-D1 using response surface methodology. Environ Sci Pollut Res Int 2014;21(9):5983-5990.##Jacob J, Rajendran RU, Priya SH, Purushothaman J, Saraswathy Amma DKBN. Enhanced antibacterial metabolite production through the application of statistical methodologies by a streptomyces nogalater NIIST A30 isolated from western ghats forest soil. PLOS One 2017;12(4):e0175919.##Managamuri U, Vijayalakshmi M, Ganduri VSRK, Rajulapati SB, Bonigala B, Kalyani BS, et al. Isolation, identification, optimization, and metabolite profiling of Streptomyces sparsus VSM-30. 3 Biotech 2017;7(3):217.##Talukdar S, Talukdar M, Buragohain M, Yadav A, Yadav RN, Bora TC. Enhanced candicidal compound production by a new soil isolate penicillium verruculosum MKH7 under submerged fermentation. BMC Microbiol 2016;16(1):288.##Kanimozhi J, Moorthy IG, Sivashankar R, Sivasubramanian V. Optimization of dextran production by Weissella cibaria NITCSK4 using Response Surface Methodology-Genetic Algorithm based technology. Carbohydr Pol 2017;174:103-110.##Wang YH, Feng JT, Zhang Q, Zhang X. Optimization of fermentation condition for antibiotic production by Xenorhabdus nematophila with response surface methodology. J Appl Microbiol 2008;104(3):735-744.##Yun TY, Feng RJ, Zhou DB, Pan YY, Chen YF, Wang F, et al. Optimization of fermentation conditions through response surface methodology for enhanced antibacterial metabolite production by Streptomyces sp. 1-14 from cassava rhizosphere. PLOS One 2018;13(11):e0206497.##

Investigation of Integron-Associated Resistance Gene Cassettes in Urinary Isolates of Klebsiella pneumoniae in Yasuj, Southwestern Iran during 2015-2016 2 2 Background: Growing antibiotic resistance among urinary opportunistic pathogens such as Klebsiella pneumoniae (K. pneumonia) has created a worrisome condition in the treatment of the Urinary Tract Infections (UTIs) in recent years. Integrons play a significant role in the dissemination of antibiotic resistance genes. The present study was conducted to investigate class 1-3 integrons and the corresponding resistance gene cassettes in urinary K. pneumoniae isolates. Methods: In this study, from December 2015 to September 2016, a total of 196 K. pneumoniae isolates were collected from the patients with UTI referred to medical diagnostic laboratories in Yasouj, Southwestern Iran. Antibiotic susceptibility patterns of isolates were determined using 12 antibiotics by the disc diffusion method. Polymerase Chain Reaction (PCR) was used for detection of integron genes (intI1, intI2, and intI3). The variable regions of integrons were amplified by PCR and sequenced to identify the corresponding gene cassettes. Results: Thirty-nine different antibiotic resistance profiles were observed among K. pneumoniae isolates. Only 12.2% of K. pneumoniae isolates were found to harbor the intI1 gene. While 17 (60.7%) out of 28 Multidrug Resistance (MDR) K. pneumoniae isolates carried the intI1 gene, only 4.2% of non-MDR isolates harbored intI1 gene. Totally 7 different gene cassette arrays were found in the intI1 gene of K. pneumoniae isolates. The aadA1 was the most prominent gene cassette. Also, high frequency of dfrA containing gene cassettes was observed. Conclusion: Continuous monitoring and characterization of integrons and their associated gene cassettes could be helpful in controlling the rising rate of antibiotic resistance. 124 131 Fariba Jahanbin Department of Basic Sciences, Islamic Azad University, Yasuj Branch Department of Basic Sciences, Islamic Azad University, Yasuj Branch Iran Masoud Marashifard Treatment Management of Social Security Organization of Kohgiluyeh and Boyer-Ahmad Province Treatment Management of Social Security Organization of Kohgiluyeh and Boyer-Ahmad Province Iran Sanaz Jamshidi Department of Basic Sciences, Islamic Azad University, Yasuj Branch Department of Basic Sciences, Islamic Azad University, Yasuj Branch Iran Maryam Zamanzadeh Department of Basic Sciences, Islamic Azad University, Yasuj Branch Department of Basic Sciences, Islamic Azad University, Yasuj Branch Iran Masumeh Dehshiri Cellular and Molecular Research Center, Yasuj University of Medical Sciences Cellular and Molecular Research Center, Yasuj University of Medical Sciences Iran Seyed Ali Asghar Malek Hosseini Student Research Committee, Yasuj University of Medical Sciences Student Research Committee, Yasuj University of Medical Sciences Iran Seyed Sajjad Khoramrooz Medicinal Plants Research Center, Yasuj University of Medical SciencesDepartment of Microbiology, Faculty of Medicine, Yasuj University of Medical Sciences Medicinal Plants Research Center, Yasuj University of Medical SciencesDepartment of Microbiology, Faculty of Medicine, Yasuj University of Medical Sciences IranIran Antibiotic resistanceIntegronsIranKlebsiella pneumoniae 20408.pdf El‐Najjar NG, Farah MJ, Hashwa FA, Tokajian ST. Antibiotic resistance patterns and sequencing of class I integron from uropathogenic Escherichia coli in Lebanon. Lett Appl Microbiol 2010;51(4):456-461.##Salem MM, Magdy M, Alhosiny IM. Distribution of classes 1 and 2 integrons among multi drug resistant E. coli isolated from hospitalized patients with urinary tract infection in Cairo, Egypt. Aust J Basic Appl Sci 2010;4(3):398-407.##Márquez C, Labbate M, Raymondo C, Fernández J, Gestal AM, Holley M, et al. Urinary tract infections in a South American population: dynamic spread of class 1 integrons and multidrug resistance by homologous and site-specific recombination. J Clin Microbiol 2008;46(10):3417-3425.##Li B, Hu Y, Wang Q, Yi Y, Woo PC, Jing H, et al. Structural diversity of class 1 integrons and their associated gene cassettes in Klebsiella pneumoniae isolates from a hospital in China. PloS One 2013;8(9):e75805.##Japoni A, Gudarzi M, Farshad S, Basiri E, Ziyaeyan M, Alborzi A, et al. Assay for integrons and pattern of antibiotic resistance in clinical Escherichia coli strains by PCR-RFLP in Southern Iran. Jpn J Infect Dis 2008;61(1):85-8.##Domingues S, da Silva GJ, Nielsen KM. Global dissemination patterns of common gene cassette arrays in class 1 integrons. Microbiology 2015;161(7):1313-1337.##Gillings MR. Integrons: past, present, and future. Microbiol Mol Biol Rev. 2014;78(2):257-277.##Stokes HW, O'gorman DB, Recchia GD, Parsekhian M, Hall RM. Structure and function of 59‐base element recombination sites associated with mobile gene cassettes. Mol Microbiol 1997;26(4):731-745.##Deng Y, Bao X, Ji L, Chen L, Liu J, Miao J, et al. Resistance integrons: class 1, 2 and 3 integrons. Ann Clin Microbiol Antimicrob 2015;14:45.##Partridge SR. Analysis of antibiotic resistance regions in Gram-negative bacteria. FEMS Microbiol Rev 2011;35(5):820-855.##Partridge SR, Tsafnat G, Coiera E, Iredell JR. Gene cassettes and cassette arrays in mobile resistance integrons. FEMS Microbiol Rev 2009;33(4):757-784.##Mazel D. Integrons: agents of bacterial evolution. Nat Rev Microbiol 2006;4(8):608-620.##DeLappe N, O'Halloran F, Fanning S, Corbett-Feeney G, Cheasty T, Cormican M. Antimicrobial resistance and genetic diversity of Shigella sonnei isolates from western Ireland, an area of low incidence of infection. J Clin Microbiol 2003;41(5):1919-1924.##Zeighami H, Haghi F, Masumian N, Hemmati F, Samei A, Naderi G. Distribution of integrons and gene cassettes among uropathogenic and diarrheagenic Escherichia coli isolates in Iran. Microb Drug Resist 2015;21(4):435-440.##Akrami F, Shahandashti EF, Yahyapour Y, Sadeghi M, Khafri S, Pournajaf A, et al. Integron types, gene cassettes and antimicrobial resistance profile of Acinetobacter baumannii isolated from BAL samples in Babol, north of Iran. Microb Pathog 2017;109:35-38.##Haghi F, Keramati N, Hemmati F, Zeighami H. Distribution of integrons and gene cassettes among metallo-β-lactamase producing Pseudomonas aeruginosa clinical isolates. Infect Epidemiol Microbiol 2017;3(2):36-40.##Akya A, Lorestani RC, Rostamian M, Elahi A, Baakhshii S, Aliabadi M, et al. The relationship of class I integron gene cassettes and the multidrug-resistance in extended-spectrum β -lactamase producing isolates of Escherichia coli. Pediatr Infect Dis J 2019;7(3).e87961.##Firoozeh F, Mahluji Z, Khorshidi A, Zibaei M. Molecular characterization of class 1, 2 and 3 integrons in clinical multi-drug resistant Klebsiella pneumoniae isolates. Antimicrob Resist Infect Control 2019;8:59.##Mahon CR, Lehman DC, Manuselis G. Textbook of diagnostic microbiology-E-Book. 6th ed. USA: Elsevier health sciences; 2014. 1024 p.##Clinical and Laboratory Standards Institute (2015) Performance standards for antimicrobial susceptibility testing. In: Twenty-Fifth informational supplement, Wayne, PA: CLSI: M100- S25.##Magiorakos AP, Srinivasan A, Carey RB, Carmeli Y, Falagas ME, Giske CG, et al. Multidrug‐resistant, extensively drug‐resistant and pandrug‐resistant bacteria: an international expert proposal for interim standard definitions for acquired resistance. Clin Microbiol Infect 2012;18(3):268-281.##Khoramrooz SS, Sharifi A, Yazdanpanah M, Malek Hosseini SA, Emaneini M, Gharibpour F, et al. High frequency of class 1 integrons in Escherichia coli isolated from patients with urinary tract infections in Yasuj, Iran. Iran Red Crescent Med J 2016;18(1):e26399.##Kraft CA, Timbury MC, Platt DJ. Distribution and genetic location of Tn7 in trimethoprim-resistant Escherichia coli. J Med Microbiol 1986;22(2):125-131.##Goldstein C, Lee MD, Sanchez S, Hudson C, Phillips B, Register B, et al. Incidence of class 1 and 2 integrases in clinical and commensal bacteria from livestock, companion animals, and exotics. 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Antimicrobial susceptibility profile of urinary isolates of Escherichia coli and Klebsiella pneumoniae. Int J Health Sci Res 2015;5(2):169-172.##Najjuka CF, Kateete DP, Kajumbula HM, Joloba ML, Essack SY. Antimicrobial susceptibility profiles of Escherichia coli and Klebsiella pneumoniae isolated from outpatients in urban and rural districts of Uganda. BMC Res Notes 2016;9(1):235.##Akram M, Shahid M, Khan AU. Etiology and antibiotic resistance patterns of community-acquired urinary tract infections in JNMC Hospital Aligarh, India. Ann Clin Microbiol Antimicrob 2007;6(1):4.##Ahangarzadeh Rezaee M, Langarizadeh N, Aghazadeh M. First report of class 1 and class 2 integrons in multidrug-resistant Klebsiella pneumoniae isolates from northwest Iran. Jpn J Infect Dis 2012;65(3):256-259.##Hendriksen RS, Munk P, Njage P, van Bunnik B, McNally L, Lukjancenko O, et al. Global monitoring of antimicrobial resistance based on metagenomics analyses of urban sewage. 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Distribution and characterization of integrons in Escherichia coli strains of animal and human origin. FEMS Immunol Med Microbiol 2007;50(1):126-132.##Gu B, Pan S, Wang T, Zhao W, Mei Y, Huang P, et al. Novel cassette arrays of integrons in clinical strains of Enterobacteriaceae in China. Int J Antimicrob Agents 2008;32(6):529-633.##Sivri N, Sandalli C, Ozgumus OB, Colakoglu F, Dogan D. Antibiotic resistance profiles of enteric bacteria isolated from Kucukcekmece Lagoon (Istanbul-Turkey). Turk J Fish Aquat Sci 2012;12(3):699-707.##

Gene Expression and Levels of TGF-B in PBMC is Associated with Severity of Symptoms in Chronic Heart Failure 2 2 Background: TGF-β1 is known to promote cardiac remodeling and fibrosis during Congestive Heart Failure (CHF). In this study, an attempt was made to investigate expression of Transforming Growth Factor beta1 (TGF-β1) and relative expansion or contraction of regulatory T-cell (Tregs) population in peripheral blood of patients with Chronic Heart Failure (CHF). Methods: Real-time PCR assay was used to investigate expression and post-stimulation levels of TGF-β1 in cell culture supernatant of Peripheral Blood Mononuclear Cells (PBMC) of 42 patients with CHF and 42 controls. Flow cytometry was used to identify relative counts of CD4+CD25+FoxP3+ Tregs. Results: PBMCs in patients with CHF expressed higher levels of TGF-β1 compared to controls. Post-stimulation levels of TGF-β1 expression were significantly higher in New York Heart Association (NYHA) functional class IV patients compared to stage I patients. Tregs were significantly expanded in PBMC in CHF, while the CD4+ helper T-cells were unchanged. Treg expansion was more significant in NYHA functional class I patients compared to class IV patients. Conclusion: Expansion of Treg population in CHF provides an extrinsic source for TGF-β1 production to induce reactive fibrosis and cardiac remodeling. Relative decrease in Treg population at advanced stages of CHF is indicative of a loss of regulatory characteristics in these cells and unopposed proinflammatory milieu.  132 134 Samaneh Saadati Department of Immunology, Faculty of Medicine, Tehran University of Medical Sciences Department of Immunology, Faculty of Medicine, Tehran University of Medical Sciences Iran Vajiheh Eskandari Cellular and Molecular Research Center, Faculty of Medicine, Guilan University of Medical Science Cellular and Molecular Research Center, Faculty of Medicine, Guilan University of Medical Science Iran Farzaneh Rahmani Research Center for Immunodeficiencies, Pediatrics Center of Excellence, Children's Medical Center, Tehran University of Medical SciencesNeuroImaging Network (NIN), Universal Scientific Education and Research Network (USERN) Research Center for Immunodeficiencies, Pediatrics Center of Excellence, Children's Medical Center, Tehran University of Medical SciencesNeuroImaging Network (NIN), Universal Scientific Education and Research Network (USERN) IranIran Mohammad Jafar Mahmoudi Department of Cardiology, Amir Alam Hospital, Tehran University of Medical Sciences Department of Cardiology, Amir Alam Hospital, Tehran University of Medical Sciences Iran Zahra Rahnemoon Cardiac Heart Center, Faculty of Medicine, Tehran University of Medical Sciences Cardiac Heart Center, Faculty of Medicine, Tehran University of Medical Sciences Iran Zahra Rahmati Department of Immunology, Faculty of Medicine, Tehran University of Medical Sciences Department of Immunology, Faculty of Medicine, Tehran University of Medical Sciences Iran Fatemeh Gorzin Department of Immunology, Faculty of Medicine, Tehran University of Medical Sciences Department of Immunology, Faculty of Medicine, Tehran University of Medical Sciences Iran Mona Hedayat Division of Immunology, Boston Children's Hospital, Harvard Medical School, Boston Division of Immunology, Boston Children's Hospital, Harvard Medical School, Boston USA Ali Akbar Amirzargar Nima Rezaei Research Center for Immunodeficiencies, Pediatrics Center of Excellence, Children's Medical Center, Tehran University of Medical SciencesNetwork of Immunity in Infection, Malignancy and Autoimmunity (NIIMA), Universal Scientific Education and Research Network (USERN), Research Center for Immunodeficiencies, Pediatrics Center of Excellence, Children's Medical Center, Tehran University of Medical Sciences IranUSA Cell culture techniquesChronic heart failureT-lymphocytesTransforming growth factor beta1 20410.pdf Glazer NL, Macy EM, Lumley T, Smith NL, Reiner AP, Psaty BM, et al. Transforming growth factor beta-1 and incidence of heart failure in older adults: the cardiovascular health study. Cytokine 2012;60(2):341-345.##Khan S, Joyce J, Margulies KB, Tsuda T. Enhanced bioactive myocardial transforming growth factor-beta in advanced human heart failure. Circ J 2014;78(11):2711-2718.##Bai Y, Zhang P, Zhang X, Huang J, Hu S, Wei Y. LTBP-2 acts as a novel marker in human heart failure - a preliminary study. Biomarkers 2012;17(5):407-415.##Edgley AJ, Krum H, Kelly DJ. Targeting fibrosis for the treatment of heart failure: a role for transforming growth factor-beta. Cardiovasc Ther 2012;30(1):e30-40.##Tulacz D, Mackiewicz U, Maczewski M, Maciejak A, Gora M, Burzynska B. Transcriptional profiling of left ventricle and peripheral blood mononuclear cells in a rat model of postinfarction heart failure. BMC Med Genomics 2013;6:49.##Eskandari V, Amirzargar AA, Mahmoudi MJ, Rahnemoon Z, Rahmani F, Sadati S, et al. Gene expression and levels of IL-6 and TNFalpha in PBMCs correlate with severity and functional class in patients with chronic heart failure. Irish J Med Sci 2018;187(2):359-368.##Rahmati Z, Amirzargar AA, Saadati S, Rahmani F, Mahmoudi MJ, Rahnemoon Z, et al. Association of levels of interleukin 17 and T-helper 17 count with symptom severity and etiology of chronic heart failure: a case-control study. Croat Med J 2018;59(4):139-148.##Schmittgen TD, Livak KJ. Analyzing real-time PCR data by the comparative C(T) method. Nat Protoc 2008;3(6):1101-1108.##Gorzin F, Amirzargar AA, Mahmoudi MJ, Rahnemoon Z, Najmi Varzaneh F, Hedayat M, et al. FOXP3, RORγt and IL-10 cytokine profile in chronic heart failure. Bratislava Lek Listy 2017;118(10):637-641.##Okamoto N, Noma T, Ishihara Y, Miyauchi Y, Takabatake W, Oomizu S, et al. Prognostic value of circulating regulatory T cells for worsening heart failure in heart failure patients with reduced ejection fraction. 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Construction, Cloning, and Expression of CagA Recombinant Protein of Helicobacter pylori 2 2 Background: This study aimed to assess construction and expression of CagA recombinant protein of Helicobacter pylori (H. pylori) in Escherichia coli (E. coli) BL21. Methods: Bioinformatics was used in designing the desired gene by Gene Runner. Next, the construct was subcloned to pET21b vector and this process was confirmed by Polymerase Chain Reaction (PCR), enzyme digestion and sequencing techniques. Then, it was cloned in the Escherichia coli BL21 as an expression host. Expression of protein was verified using sodium dodecyl sulfate- polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting technique. For purification of the protein, the Ni-NTA column was used. Protein concentration was determined by the Bicinchoninic Acid Protein Assay Kit (Parstoos). Finally, Western blotting was performed using CagA antibodies and normal human serum for determining immunogenicity feature with human antiserum. Results: According to the results of the present study, CagA construct was cloned into the pET21b vector and after confirmation and cloning in host expression, recombinant protein with the size of 38 kDa was successfully expressed and purified. The recombinant CagA protein showed immunogenicity characteristics with human antiserum. Conclusion: In conclusion, only 5′-end of recombinant protein CagA with high immunogenicity effects was successfully constructed, cloned and expressed.  Also, CagA recombinant protein showed good immunogenicity activity with human antiserum. 135 138 Abbas Shapouri Moghaddam Department of Immunology, BuAli Research Institute, Faculty of Medicine, Mashhad University of Medical Sciences Department of Immunology, BuAli Research Institute, Faculty of Medicine, Mashhad University of Medical Sciences Iran Shamseddin Mansouri Antimicrobial Resistance Research Center, Department of Microbiology, Ghaem Hospital, Mashhad University of Medical Sciences Antimicrobial Resistance Research Center, Department of Microbiology, Ghaem Hospital, Mashhad University of Medical Sciences Iran Alireza Neshani Antimicrobial Resistance Research Center, Department of Microbiology, Ghaem Hospital, Mashhad University of Medical Sciences Antimicrobial Resistance Research Center, Department of Microbiology, Ghaem Hospital, Mashhad University of Medical Sciences Iran Farzaneh Firoozeh Department of Microbiology, Faculty of Medicine, Alborz University of Medical Sciences Department of Microbiology, Faculty of Medicine, Alborz University of Medical Sciences Iran Azade Matinpur Infectious Diseases Research Center, Faculty of Medicine, Kashan University of Medical SciencesDepartment of Microbiology and Immunology, Faculty of Medicine, Kashan University of Medical Sciences Infectious Diseases Research Center, Faculty of Medicine, Kashan University of Medical SciencesDepartment of Microbiology and Immunology, Faculty of Medicine, Kashan University of Medical Sciences IranIran Azad Khaledi Mehran Ghazalibina Department of Microbiology, Faculty of Public Health, Tehran University of Medical Sciences Department of Microbiology, Faculty of Public Health, Tehran University of Medical Sciences Iran CagAHelicobacter pyloriRecombinant proteinsVaccine candidate 10375.pdf Khaledi A, Bahador A, Esmaeili D, Derakhshan M, Mahdavi M, Amani J, et al. 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