Monkeypox Outbreaks in Non-Endemic Countries: What Do We Know and What Do We Need? 2 2 Following the news of the outbreak of the monkeypox virus in non-endemic countries from the media, a new wave of concerns was created. The current susceptibility to epidemics has resulted from successive peaks of the whole-society-inclusive coronavirus and its significant mortality and morbidity. To date, on May 28, 2022, there are 401 confirmed and 85 suspected cases worldwide 1, almost all of which are from countries with high-income settings. According to Global.health, 3 suspicious and no confirmed cases have been tracked from Iran. The human monkeypox is a viral zoonosis from the genus Orthopoxvirus in the family Poxviridae that causes flu-like symptoms, including fever, fatigue, and body aches, along with progressive macular-papular, vesicular, pustular, and crusted scab skin lesions that can be itchy 2. The virus is closely related to the Variola virus, which causes smallpox in humans, but it leads to less severe symptoms and often improves without treatment; however, due to the broad spectrum of the disease severity, it can even be fatal. Previous observations reported fatality rates of 1 to 11%, which appear to be related to the virus clade, patients' age, and concomitant infection with human immunodeficiency virus 2,3. This disease is not new, and there is good, but not enough, research and background information about it. The source of the current outbreak is unknown, and scientists are looking for reds to explain its outbreak. Transmission of the disease from animals, humans, including nosocomial and household transmission, and fomites is possible 4. Like many other viral diseases, there is no definitive cure; however, antiviral drugs, like tecovirimat, vaccines, and immunoglobulins are available in developed countries, which help control complications, transmission, and epidemics 5. The smallpox vaccine is about 85% effective against the monkeypox virus and reduces the frequency and severity of its signs and symptoms. It should be noted that non-elderly people were born after the cessation of smallpox vaccination 3,6, and this may justify the storage of vaccines by developed countries. Scientists have previously reported the risks of cessation of smallpox vaccination, including the establishment and propagation of monkeypox, although they have outweighed the benefits due to the complications and costs of vaccination 3,4. Information on the source and extent of the current cases and outbreaks is insufficient. In addition, although only two strains of the virus have been identified in the past, the possibility of novel strains emerging should be considered. Multiple outbreaks, 2022 outbreaks in non-endemic countries, and the high-consequence nature of the pathogen necessitate significant improvements in the quantity and quality of data collection, basic and clinical science research, providing vaccines, people and medical staff informing and education, and guideline development. These lead to more proper management of disease cases and the promotion of public health. 186 187 Ahmad Shamabadi School of Medicine, Tehran University of Medical SciencesPsychiatric Research Center, Roozbeh Psychiatric Hospital, Tehran University of Medical Sciences School of Medicine, Tehran University of Medical SciencesPsychiatric Research Center, Roozbeh Psychiatric Hospital, Tehran University of Medical Sciences IranIran Shahin Akhondzadeh Psychiatric Research Center, Roozbeh Psychiatric Hospital, Tehran University of Medical Sciences Psychiatric Research Center, Roozbeh Psychiatric Hospital, Tehran University of Medical Sciences Iran EpidemicsIranMass active immunizationMonkey poxVaccinesViral infection 60503.pdf Global.health: a Data Science Initiative. Monkeypox Tracker. Available online: https://monkeypox.healthmap.org (accessed on 2022 May 28).##Adler H, Gould S, Hine P, Snell LB, Wong W, Houlihan CF, et al. Clinical features and management of human monkeypox: a ret-rospective observational study in the UK. Lancet Infect Dis 2022;S1473-3099(22)00228-6.##Beer EM, Rao VB. A systematic review of the epidemiology of human monkeypox outbreaks and implications for outbreak strate-gy. PLoS Negl Trop Dis 2019;13(10):e0007791. ##Moore ZS, Seward JF, Lane JM. Smallpox. Lancet 2006;367(9508):425-35. ##Russo AT, Grosenbach DW, Chinsangaram J, Honeychurch KM, Long PG, Lovejoy C, et al. An overview of tecovirimat for smallpox treatment and expanded anti-orthopoxvirus applications. Expert Rev Anti Infect Ther 2021;19(3):331-44. ##Reynolds MG, Damon IK. Outbreaks of human monkeypox after cessation of smallpox vaccination. Trends Microbiol 2012; 20(2):80-7. ##

Expression of Toll-Like Receptors 2, 4 and 5 in Relation to Gut Microbiota in Colon Neoplasm Patients with and without Inflammatory Bowel Disease 2 2 <h1 style="text-align:justify">Background: Toll-Like Receptors (TLRs) are the critical mediators of inflammatory routs in the gut, which play an essential role in regulating the immune responses towards various ligands derived from pathogenic bacteria. Also, TLR signaling has been implicated in the development of Inflammatory Bowel Disease (IBD), Adenomatous Polyp (AP), and Colorectal Cancer (CRC). Here, we aimed to examine the expression of some TLRs concerning certain fecal bacteria in AP and CRC patients with and without IBD.</h1> Methods: This case-control study collected fecal and colonic tissue samples from 93 patients versus Normal Controls (NC) via colonoscopy. Fecal samples were used for DNA extraction, and the abundance of selected fecal bacteria was determined by absolute real-time PCR. Also, the gene expression of TLR2, 4, and 5 was analyzed using RT-PCR on the colonic tissues of participants. Results: Compared to NC individuals, in AP and CRC patients, the mRNA expressions of TLR4 and TLR2 were significantly increased while TLR5 was decreased. A meaningful association between TLRs mRNA expression levels and the abundance of some selected fecal bacteria was detected. Also, there was a significant relationship between participant’s food regimes, smoking habit and intestinal TLRs expression. Conclusion: Our study proposed the important role of TLRs during adenomatous and CRC formation. Alterations in TLRs expression associated with certain gut bacteria may contribute to disease development. 188 195 Hamid Asadzadeh Aghdaei Basic and Molecular Epidemiology of Gastrointestinal Disorders Research Center, Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical Sciences Iran Sama Rezasoltani Foodborne and Waterborne Diseases Research Center, Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical Sciences Iran Meisam Olfatifar Gastroenterology and Liver Diseases Research Center, Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical Sciences Iran Ehsan Nazemalhosseini Mojarad Gastroenterology and Liver Diseases Research Center, Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical Sciences Iran Ghazal Sherkat Basic and Molecular Epidemiology of Gastrointestinal Disorders Research Center, Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical Sciences Iran Abbas Yadegar Foodborne and Waterborne Diseases Research Center, Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical Sciences Iran Mohammad Mehdi Feizabadi Department of Microbiology, School of Medicine, Tehran University of Medical Sciences Department of Microbiology, School of Medicine, Tehran University of Medical Sciences Iran Mohammad Reza Zali Gastroenterology and Liver Diseases Research Center, Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical Sciences Iran Adenomatous polypColorectal cancerFecal bacteriaInflammatory bowel diseaseToll-like receptors 60504.pdf Kuipers EJ, Grady WM, Lieberman D, Seufferlein T, Sung JJ, Boelens PG, et al. Colorectal cancer. Nat Rev Dis Primers 2015;1:15065. ##Rezasoltani S, Ghanbari R, Looha MA, Mojarad EN, Yadegar A, Stewart D, et al. Expression of main toll-like receptors in patients with different types of colorectal polyps and their relationship with gut microbiota. Int J Mol Sci 2020;21(23):8968. ##Haggar FA, Boushey RP. Colorectal cancer epidemiology: incidence, mortality, survival, and risk factors. Clin Colon Rectal Surg 2009;22(4):191-7. ##Worthley DL, Leggett BA. Colorectal cancer: molecular features and clinical opportunities. Clin Biochem Rev 2010;31(2):31-8. ##Lao VV, Grady WM. Epigenetics and colorectal cancer. Nat Rev Gastroenterol Hepatol 2011;8(12):686-700. ##Wanders LK, Dekker E, Pullens B, Bassett P, Travis SP, East JE. Cancer risk after resection of polypoid dysplasia in patients with longstanding ulcerative colitis: a meta-analysis. Clin Gastroenterol Hepatol 2014;12(5):756-64. ##Stidham RW, Higgins PDR. Colorectal cancer in inflam-matory bowel disease. Clin Colon Rectal Surg 2018; 31(3):168-78. ##Yang T, Owen JL, Lightfoot YL, Kladde MP, Moham-adzadeh M. Microbiota impact on the epigenetic regulation of colorectal cancer. Trends Mol Med 2013;19(12): 714-25. ##Sobhani I, TranVanNhieu J. Colon cancer is associated with microbial dysbiosis in humans and animals. Go-varesh 2017;18(1):45-56.##Sekirov I, Russell SL, Antunes LC, Finlay BB. Gut mi-crobiota in health and disease. Physiol Rev 2010;90(3): 859-904. ##Yesudhas D, Gosu V, Anwar MA, Choi S. Multiple roles of toll-like receptor 4 in colorectal cancer. Front Immunol 2014;5:334. ##Spiljar M, Merkler D, Trajkovski M. The immune system bridges the gut microbiota with systemic energy homeo-stasis: focus on TLRs, mucosal barrier, and SCFAs. Front Immunol 2017;8:1353. ##Lavelle EC, Murphy C, O’Neill LAJ, Creagh EM. The role of TLRs, NLRs, and RLRs in mucosal innate im-munity and homeostasis. Mucosal Immunol 2010;3(1): 17-28. ##Navegantes KC, de Souza Gomes R, Pereira PAT, Czai-koski PG, Azevedo CHM, Monteiro MC. Immune modu-lation of some autoimmune diseases: the critical role of macrophages and neutrophils in the innate and adaptive immunity. J Transl Med 2017;15(1):36. ##Messaritakis I, Stogiannitsi M, Koulouridi A, Sfakianaki M, Voutsina A, Sotiriou A, et al. Evaluation of the detec-tion of Toll-like receptors (TLRs) in cancer development and progression in patients with colorectal cancer. PLoS One 2018;13(6):e0197327. ##Paarnio K, Väyrynen S, Klintrup K, Ohtonen P, Mäkinen MJ, Mäkelä J, et al. Divergent expression of bacterial wall sensing Toll-like receptors 2 and 4 in colorectal cancer. World J Gastroenterol 2017;23(26):4831-8. ##Yiu JH, Dorweiler B, Woo CW. Interaction between gut microbiota and toll-like receptor: from immunity to metab-olism. J Mol Med (Berl) 2017;95(1):13-20. ##Francescone R, Hou V, Grivennikov SI. Microbiome, inflammation, and cancer. Cancer J 2014;20(3):181-9. ##Brennan CA, Garrett WS. Gut Microbiota, Inflammation, and Colorectal Cancer. Annu Rev Microbiol 2016;70: 395-411. ##Rezasoltani S, Sharafkhah M, Asadzadeh Aghdaei H, Nazemalhosseini Mojarad E, Dabiri H, Akhavan Sepahi A, et al. Applying simple linear combination, multiple lo-gistic and factor analysis methods for candidate fecal bac-teria as novel biomarkers for early detection of adenoma-tous polyps and colon cancer. J Microbiol Methods 2018;155:82-8. ##Rezasoltani S, Asadzadeh Aghdaei H, Dabiri H, Akhavan Sepahi A, Modarressi MH, Nazemalhosseini Mojarad E. The association between fecal microbiota and different types of colorectal polyp as precursors of colorectal can-cer. Microb Pathog 2018;124:244-9. ##Kim TW, Lee SJ, Oh BM, Lee H, Uhm TG, Min JK, et al. Epigenetic modification of TLR4 promotes activation of NF-κB by regulating methyl-CpG-binding domain protein 2 and Sp1 in gastric cancer. Oncotarget 2016;7 (4):4195-209. ##Li H, Yang T, Li FY, Ning Q, Sun ZM. TLR4 Overex-pression inhibits endothelial PAS domain containing pro-tein 1 expression in the lower respiratory tract of patients with chronic COPD. Cell Physiol Biochem 2016;39(2): 685-92. ##Peyravian N, Gharib E, Moradi A, Mobahat M, Tarban P, Azimzadeh P, et al. Evaluating the expression level of co-stimulatory molecules CD 80 and CD 86 in different types of colon polyps. Curr Res Transl Med 2018;66(1): 19-25. ##Aziz Q, Doré J, Emmanuel A, Guarner F, Quigley EM. Gut microbiota and gastrointestinal health: current con-cepts and future directions. Neurogastroenterol Motil 2013;25(1):4-15. ##Maranduba CM, De Castro SB, de Souza GT, Rossato C, da Guia FC, Valente MA, et al. Intestinal microbiota as modulators of the immune system and neuroimmune sys-tem: impact on the host health and homeostasis. J Immu-nol Res 2015; 2015:931574. ##Rezasoltani S, Asadzadeh-Aghdaei H, Nazemalhosseini-Mojarad E, Dabiri H, Ghanbari R, Zali MR. Gut micro-biota, epigenetic modification and colorectal cancer. Iran J Microbiol 2017;9(2):55-63. ##Kutikhin AG, Yuzhalin AE, Tsitko EA, Brusina EB. Pattern recognition receptors and DNA repair: starting to put a jigsaw puzzle together. Front Immunol 2014;5:343. ##Kelly D, Mulder IE. Microbiome and immunological in-teractions. Nutr Rev 2012;70 Suppl 1:S18-30. ##Mogensen TH. Pathogen recognition and inflammatory signaling in innate immune defenses. Clin Microbiol Rev 2009;22(2):240-73. ##Lee YK, Mehrabian P, Boyajian S, Wu WL, Selicha J, Vonderfecht S, et al. The protective role of Bacteroides fragilis in a murine model of colitis-associated colorectal cancer. mSphere 2018 14;3(6):e00587-18. ##Li R, Zhou R, Wang H, Li W, Pan M, Yao X, et al. Gut microbiota-stimulated cathepsin K secretion mediates TLR4-dependent M2 macrophage polarization and pro-motes tumor metastasis in colorectal cancer. Cell Death Differ 2019;26(11):2447-63. ##Rezasoltani S, Khatibi S, Pezeshkiyan Z, Nazemalhos-seini-Mojarad E, Sharafkhah M, Sadeghi A, et al. Investi-gating the TLR9 mRNA expression level in different his-tological types of colorectal polyps. Asian Pac J Cancer Prev 2019;20(8):2299-302. ##Shukla R, Ghoshal U, Ranjan P, Ghoshal UC. Expression of toll-like receptors, pro-, and anti-inflammatory cyto-kines in relation to gut microbiota in irritable bowel syn-drome: The evidence for its micro-organic basis. J Neuro-gastroenterol Motil 2018;24(4):628-42. ##Inoue R, Yajima T, Tsukahara T. Expression of TLR2 and TLR4 in murine small intestine during postnatal develop-ment. Biosci Biotechnol Biochem 2017;81(2):350-8. ##

Genome Analysis of an Enterococcal Prophage, Entfac.MY 2 2 Background: Bacteriophages are bacterial parasites. Unlike lytic bacteriophages, lysogenic bacteriophages do not multiply immediately after entering the host cells and may integrate their genomes into the bacterial genomes as prophages. Prophages can include various phenotypic and genotypic effects on the host bacteria. Enterococcus spp. are Gram-positive bacteria that cause infections in humans and animals. In recent decades, these bacteria have become resistant to various antimicrobials, including vancomycin. The aim of this study was to analyze genome of an enterococcal prophage. Methods: In this study, Enterococcus faecium EntfacYE was isolated from biological samples and its genome was analyzed using next-generation sequencing method. Results: Overall, 254 prophage genes were identified in the bacterial genome. The prophage included 39 housekeeping, 41 replication and regulation, 80 structural and packaging, and 48 lysis genes. Moreover, 46 genes with unknown functions were identified. All genes were annotated in DNA Data Bank of Japan. Conclusion: In general, most prophage genes were linked to packaging and structure (31.5%) gene group. However, genes with unknown functions included a high proportion (18.11%), which indicated necessity of further analyses. Genomic analysis of the prophages can be effective in better understanding of their roles in development of bacterial resistance to antibiotics. Moreover, identification and study of prophages can help researchers develop genetic engineering tools and novel infection therapies. 196 205 Maryam Yazdanizad Department of Medical Biotechnology, Faculty of Advanced Science and Technology, Tehran Medical Sciences, Islamic Azad University Department of Medical Biotechnology, Faculty of Advanced Science and Technology, Tehran Medical Sciences, Islamic Azad University Iran Ramin Mazaheri Nezhad Fard Department of Pathobiology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran 3. Food Microbiology Research Center, Tehran University of Medical SciencesFood Microbiology Research Center, Tehran University of Medical Sciences Department of Pathobiology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran 3. Food Microbiology Research Center, Tehran University of Medical Sciences IranIran Golshid Javdani Shahedin Pasteur Institute of Iran Iran Mohammadreza Salehi Department of Infectious Diseases, School of Medicine, Tehran University of Medical Sciences Department of Infectious Diseases, School of Medicine, Tehran University of Medical Sciences Iran Mahsa Dumanloo Department of Clinical Pathobiology, Imam Khomeini Hospital Complex, Tehran University of Medical Sciences Iran Ali Akbar Saboor Yaraghi Department of Pathobiology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran 3. Food Microbiology Research Center, Tehran University of Medical Sciences Department of Pathobiology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran 3. Food Microbiology Research Center, Tehran University of Medical Sciences Iran Enterococcus faeciumGenomic analysisProphages 60505.pdf Fisher K, Phillips C. The ecology, epidemiology and viru-lence of Enterococcus. Microbiology 2009;155(Pt 6):1749-57. ##Garcia-Solache M, Rice LB. The Enterococcus: a model of adaptability to its environment Clin Microbiol Rev 2019;32(2):e00058-18. ##Zaheer R, Cook SR, Barbieri R, Goji N, Cameron A, Petkau A, et al. Surveillance of Enterococcus spp. reveals distinct species and antimicrobial resistance diversity across a One-Health continuum. Sci Rep 2020;10(1): 3937. ##Topka-Bielecka G, Bloch S, Nejman-Falenczyk B, Grab-ski M, Jurczak-Kurek A, Gorniak M, et al. Characteriza-tion of the bacteriophage vB_EfaS-271 infecting Entero-coccus faecalis. Int J Mol Sci 2020;21(17):6345. ##Revtovich AV, Tjahjono E, Singh KV, Hanson BM, Mur-ray BE, Kirienko NV. Development and characterization of high-throughput Caenorhabditis elegans–Enter-ococcus faecium infection model. Front Cell Infect Microbiol 2021;11:667327. ##Abdulrahman Ashy R, Agusti S. Low host abundance and high temperature determine switching from lytic to lyso-genic cycles in planktonic microbial communities in a trop-ical sea (Red Sea). Viruses 2020;12(7):761. ##Stepien-Pysniak D, Hauschild T, Dec M, Marek A, Ur-ban-Chmiel R, Kosikowska U. Phenotypic and genotypic characterization of Enterococcus spp. from yolk sac infec-tions in broiler chicks with a focus on virulence factors. Poult Sci 2021;100(4):100985. ##Nasiri M, Hanifian S. Enterococcus faecalis and Entero-coccus faecium in pasteurized milk: Prevalence, genotyp-ing, and characterization of virulence traits. LWT 2022; 153:112452.##Elahi Y, Nowroozi J, Fard RMN. Isolation and characteri-zation of bacteriophages from wastewater sources on En-terococcus spp. isolated from clinical samples. Iran J Mi-crobiol 2021;13(5):671-7. ##Fard RM, Barton MD, Arthur JL, Heuzenroeder MW. Whole-genome sequencing and gene mapping of a newly isolated lytic enterococcal bacteriophage EFRM31. 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J Am Chem Soc 2018;140(49):17153-62. ##Lee SJ, Zhu B, Hamdan SM, Richardson CC. Mechanism of sequence-specific template binding by the DNA pri-mase of bacteriophage T7. Nucleic Acids Res 2010; 38(13):4372-83. ##Enriquez A, Chu IW, Mellis C, Lin WY. Nebulised deox-yribonuclease for viral bronchiolitis in children youn-ger than 24 months. Cochrane Database Syst Rev 2012; (11): 11(11):CD008395. ##Voziyanova E, Li F, Shah R, Voziyanov Y. Genome tar-geting by hybrid Flp-TAL recombinases. Sci Rep 2020;10(1):17479. ##Silpe JE, Bridges AA, Huang X, Coronado DR, Duddy OP, Bassler BL. Separating functions of the phage-encoded quorum-sensing-activated antirepressor Qtip. Cell Host Microbe 2020;27(4):629-41. ##Skerry C, Goldman WE, Carbonetti NH. Peptidoglycan recognition protein 4 suppresses early inflammatory re-sponses to Bordetella pertussis and contributes to sphin-gosine-1-phosphate receptor agonist-mediated disease at-tenuation. Infect Immun 2019;87(2):e00601-18. ##Jiang L, Liu W, Guo H, Dang Y, Cheng T, Yang W, et al. Distinct functions of Bombyx mori peptidoglycan recogni-tion protein 2 in immune responses to bacteria and viruses. Front Immunol 2019;10:776. ##Stojkova P, Spidlova P, Stulik J. Nucleoid-associated protein HU: A lilliputian in gene regulation of bacterial virulence. Front Cell Infect Microbiol 2019;9:159. ##Furuta Y, Namba-Fukuyo H, Shibata TF, Nishiyama T, Shigenobu S, Suzuki Y, et al. Methylome diversification through changes in DNA methyltransferase sequence specificity. PLoS Genet 2014;10(4):e1004272. ##Stoddard CI, Feng S, Campbell MG, Liu W, Wang H, Zhong X, et al. A nucleosome bridging mechanism for ac-tivation of a maintenance DNA methyltransferase. Mol Cell 2019;73(1):73-83. ##Bardina C, Colom J, Spricigo DA, Otero J, Ssnchez-Osuna M, Cortes P, et al. Genomics of three new bacteri-ophages useful in the biocontrol of Salmonella. Front Mi-crobiol 2016;7:545. ##Stamereilers C, Fajardo CP, Walker JK, Mendez KN, Castro-Nallar E, Grose JH, et al. Genomic analysis of 48 Paenibacillus larvae bacteriophages. Viruses 2018;10 (7):377. ##Jacobs-Sera D, Abad LA, Alvey RM, Anders KR, Aull HG, Bhalla SS, et al. Genomic diversity of bacteriophages infecting Microbacterium spp. PLoS One 2020;15 (6):e0234636. ##Pawlowski A, Moilanen AM, Rissanen IA, Määttä JA, Hytonen VP, Ihalainen JA, et al. The minor capsid protein VP11 of thermophilic bacteriophage P23-77 facilitates vi-rus assembly by using lipid-protein interactions. J Virol 2015;89(15):7593-603. ##Li X, Koc C, Kuhner P, Stierhof YD, Krismer B, Enright MC, et al. An essential role for the baseplate protein Gp45 in phage adsorption to Staphylococcus aureus. Sci Rep 2016;6:26455. ##McNulty R, Lokareddy RK, Roy A, Yang Y, Lander GC, Heck AJ, et al. Architecture of the complex formed by large and small terminase subunits from bacteriophage P22. 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Isolation, Molecular Identification and Antibacterial Potential of Marine Bacteria from Deep Atlantic Ocean of Morocco 2 2 Background: Antibiotic resistance is an important concern for the public health authorities at global level. It is detrimental to human and environmental ecosystems, thus, there is a big need for natural bioactive compounds. In this work, we aimed to find out biomolecules derived from marine bacteria that may constitute an alternative to antibiotics. Methods: We isolated and identified thirty one marine bacteria collected from deep ocean water in central coast of Safi city, Morocco. Then, we induced biomolecules production in six marine bacterial strains. The extracts were tested for their antibacterial activity against gram-negative and gram-positive bacteria such as Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 33592 and Listeria monocytogenes ATCC 19117. Furthermore, we partially analyzed the chemical composition of these biomolecules and evaluated their sensibility to different temperatures. Results: The six marine bacteria were able to produce molecules which inhibited the three pathogenic strains with high inhibition zones reaching 27 mm. These molecules were characterized by heat stability from 60 to 121°C relying on each strain. Conclusion: The produced molecules may offer a great potential to pharmaceutical industries as they may constitute an alternative to antibiotics that are becoming less effective due to the emergence of drugs resistance. 206 215 Asmaa Chbel Laboratory of Physiopathology, Molecular Genetics & Biotechnology, Faculty of Sciences Ain Chock, Research Center of Health & Biotechnology, Hassan II University of Casablanca Morocco Jorge Rodriguez-Castro Laboratory of Molecular Systematics, Department of Biochemistry & Molecular Biology, CIQUS University of Santiago de Compostela Spain Javier Quinteiro Laboratory of Molecular Systematics, Department of Biochemistry & Molecular Biology, CIQUS University of Santiago de Compostela Spain Manuel Rey-Méndez Laboratory of Molecular Systematics, Department of Biochemistry & Molecular Biology, CIQUS University of Santiago de Compostela Spain Aurelio Serrano Delgado Institute of Plant Biochemistry and Photosynthesis (IBVF), CSIC-University of Sevilla Spain Abdelaziz Soukri Laboratory of Physiopathology, Molecular Genetics & Biotechnology, Faculty of Sciences Ain Chock, Research Center of Health & Biotechnology, Hassan II University of Casablanca Morocco Bouchra El Khalfi Laboratory of Physiopathology, Molecular Genetics & Biotechnology, Faculty of Sciences Ain Chock, Research Center of Health & Biotechnology, Hassan II University of Casablanca Morocco Antibacterial activityAntibiotic resistanceMarine bacteriaMarine biomolecules 60506.pdf Romano G, Costantini M, Sansone C, Lauritano C, Ruoc-co N, Ianora A. 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A New Specific DNA Target Sequence for Identification of Staphylococcus epidermidis using Modified Comparative Genomic Analysis 2 2 Background: Staphylococcus epidermidis (S. epidermidis) is the most frequently isolated pathogen from prostheses infections in the body. Therefore, improving its diagnostic methods, including rapid Nucleic Acid Amplification Tests (NAAT), seems necessary. Since the first step in designing a NAAT is to find a specific sequence and all DNA targets that have been introduced so far are not completely specific, we introduced a new 100% specific DNA target sequence to identify S. epidermidis in this study. Methods: Modified comparative genomic analysis was used to find the best specific target sequence to detect S. epidermidis. A PCR method was designed for the evaluation of this target. To determine the detection limit and analytical specificity, pure genomic DNA of 18 bacteria include 12 standard strains (one S. epidermidis and 11 non-S. epidermidis) and six clinical isolates (five S. epidermidis and one non-S. epidermidis) were used. Results: The 400 bp sequence of S. epidermidis ATCC 14990 was identified as the most specific sequence (Se400), having a 100% sequence similarity to S. epidermidis genomes but not with other bacteria. The detection limit of Se400-PCR was 10 fg, equal to about 4 copies of S. epidermidis genomic DNA/μl. All pure DNA templates from S. epidermidis generated a detectable amplicon by 264 bp length, but the PCR test was negative for the non-S. epidermidis group. Conclusion: The Se400 sequence can be considered as a specific target for detecting S. epidermidis, based on our findings. 216 222 Reza Khoshbakht Antimicrobial Resistance Research Center, Mashhad University of Medical SciencesStudent Research Committee, Mashhad University of Medical Sciences Antimicrobial Resistance Research Center, Mashhad University of Medical SciencesStudent Research Committee, Mashhad University of Medical Sciences IranIran Hosna Zare Antimicrobial Resistance Research Center, Mashhad University of Medical SciencesStudent Research Committee, Mashhad University of Medical Sciences Student Research Committee, Mashhad University of Medical Sciences IranIran Reza Kamali Kakhki Antimicrobial Resistance Research Center, Mashhad University of Medical SciencesStudent Research Committee, Mashhad University of Medical Sciences Student Research Committee, Mashhad University of Medical Sciences IranIran Alireza Neshani Student Research Committee, Mashhad University of Medical SciencesDepartment of Laboratory Sciences, School of Paramedical Sciences, Mashhad University of Medical Sciences Student Research Committee, Mashhad University of Medical SciencesDepartment of Laboratory Sciences, School of Paramedical Sciences, Mashhad University of Medical Sciences IranIran Maryam Arfaatabar Department of Medical Laboratory Sciences, Kashan Branch, Islamic Azad University Department of Medical Laboratory Sciences, Kashan Branch, Islamic Azad University Iran Comparative genomic analysisDetectionPathogenPolymerase chain reaction Se400Staphylococcus epidermidis 60507.pdf Morse SA, Mietzner TA, Miller S, Riedel S. 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GC/MS Analysis and Phyto-synthesis of Silver Nanoparticles Using Amygdalus spinosissima Extract: Antibacterial, Antioxidant Effects, Anticancer and Apoptotic Effects 2 2 Background: The present study was aimed at phyto-synthesized silver nanoparticles (AgNPs) using Amygdalus spinosissima (A. spinosissima) extract and to investigate the antibacterial, antioxidant effects, anticancer and apoptotic effects of phyto-synthe-sized AgNPs. Methods: The bio-fabricated AgNPs were characterized using UV-visible spectroscopy (UV-visible), X-ray Diffraction (XRD), Fourier Transform Infrared (FTIR), Transmission Electron Microscopy (TEM), Scanning Electron Microscopy (SEM) and Energy Disper-sive X-ray (EDX). Results: The phyto-synthesized AgNPs showed maximum absorption in 438 nm, in the UV-visible spectrum. XRD peaks were observed at 2θ values in 38.20°, 44.40°, 64.60°, and 77.50° which are indexed as (111), (200), (220), and (311) bands of Face-Centered Cubic (FCC) structures of silver. FTIR analysis indicated that the AgNPs were capped with A. spinosissima extract. SEM and TEM micrographs revealed that the fabricated AgNPs were spherical and the average size range was 17.89 nm. Also, the EDX results show that the content of Ag was 90%. Conclusion: The phyto-synthesized AgNPs had significant antibacterial activity against Gram-negative bacteria, as well as, the AgNPs exhibited great inhibitory effects on DPPH radicals and their antioxidant properties were favorably comparable to the antioxidant outcomes of ascorbic acid. Moreover, the AgNPs showed anti-cancer activity against the MCF-7 cell line with the IC50=6.1 µg/ml. Moreover, the phyto-synthesized AgNPs could induce apoptosis in the MCF-7 cell line significantly. The GC-MS analysis of the A. spinosissima extract showed that 102 bioactive phyto-chemical compounds, which be of use to the synthesis of AgNPs.     223 232 Azadeh Farmahini Farahani Department of Biology, Central Tehran branch, Islamic Azad University Department of Biology, Central Tehran branch, Islamic Azad University Iran Seyed Mohammad Mahdi Hamdi Department of Biology, Central Tehran branch, Islamic Azad University Department of Biology, Central Tehran branch, Islamic Azad University Iran Amir Mirzaee Department of Biology, Roudehen branch, Islamic Azad University Department of Biology, Roudehen branch, Islamic Azad University Iran Amygdalus spinosissimaAnti-bacterial agentsAntioxidantsApoptosisSilver nanoparticle 60508.pdf De Jong WH, Borm PJA. Drug delivery and nanoparti-cles: applications and hazards. Int J Nanomedicine 2008; 3(2):133-49. ##Jătariu A, Peptu C, Popa M, Indrei A. Micro- and nano-particles-medical applications. 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Green synthesis of silver nanopar-ticles via Cynara scolymus leaf extracts: The characteriza-tion, anticancer potential with photodynamic therapy in MCF7 cells. PLoS One 2019;14(6):e0216496. ##Sathishkumar P, Vennila K, Jayakumar R, Yusoff AR, Hadibarata T, Palvannan P, et al. Phyto-synthesis of silver nanoparticles using Alternanthera tenella leaf extract: An effective inhibitor for the migration of human breast ade-nocarcinoma (MCF-7) cells. Bioprocess Biosyst Eng 2016;39:651-9.##Manosalva N, Tortella G, Cristina Diez M, Schalchli H, Seabra AB, Durán N, et al. World J Microbiol Biotechnol 2019 May 27;35(6):88. ##Masum MMI, Siddiqa MM, Ali KA, Zhang Y, Abdallah Y, Ibrahim E, et al. Biogenic synthesis of silver nanoparti-cles using Phyllanthus emblica fruit extract and its inhibi-tory action against the pathogen Acidovorax oryzae strain RS-2 of rice bacterial brown stripe. Front Microbiol 2019;10:820. ##Salari S, Esmaeilzadeh Bahabadi S, Samzadeh-Kermani A, Yosefzaei F. In-vitro evaluation of antioxidant and anti-bacterial potential of green synthesized silver nanoparticles using Prosopis farcta fruit extract. Iran J Pharm Res 2019;18(1):430-455. ##Vallverdú-Queralt A, Regueiro J, Alvarenga JFR, Mar-tinez-Huelamo M, Leal LN, Lamuela-Raventos RM. Characterization of the phenolic and antioxidant profiles of selected culinary herbs and spices: caraway, turmeric, dill, marjoram and nutmeg. Food Sci Technol 2015;35 (1):189-95.##Prunus amygdalus Batsch. Plants of the World Online. Royal Botanic Gardens, Kew. Retrieved 1 April 2019.##BĀDĀM–Encyclopaedia Iranica. www.iranicaonline. org. Retrieved 25 May 2019. The Amygdalus communis (or Prunus amygdalus), though undoubtedly native to the Ira-nian land-mass, is seldom found in natural stands there to-day. ##Bolling BW. Almond polyphenols: methods of analysis, contribution to food quality, and health promotion. Com-prehensive Reviews Food Science Food Safety 2017;16: 346-68.##Suman TY, Rajasree SR, Jayaseelan C, Mary RR, Ga-yathri S, Aranganathan L, et al. GC-MS analysis of bioac-tive components and biosynthesis of silver nanoparticles using Hybanthus enneaspermus at room temperature eval-uation of their stability and its larvicidal activity. Environ Sci Pollut Res Int 2016 Feb;23(3):2705-14. ##Manosalva N, Tortella G, Cristina Diez M, Schalchli H, Seabra AB, Durán N, et al. Green synthesis of silver na-noparticles: effect of synthesis reaction parameters on an-timicrobial activity. World J Microbiol Biotechnol 2019; 35(6):88. ##Lee YJ, Song K, Cha SH, Cho S, Kim YS, Park Y. Ses-quiterpenoids from Tussilago farfara flower bud extract for the eco-friendly synthesis of silver and gold nanoparti-cles possessing antibacterial and anticancer activities. Na-nomaterials (Basel) 2019;9(6):819. ##Al-Sheddi ES, Farshori NN, Al-Oqail MM, Al-Massarani SM, Saquib Q, Bioinorg R. Wahab R, et al. Anticancer potential of green synthesized silver nanoparticles using extract of Nepeta deflersiana against human cervical cancer cells (HeLA). Bioinorg Chem Appl 2018;2018: 9390784. ##Baharara J, Namvar F, Ramezani T, Mousavi M, Moham-ad R. Silver nanoparticles biosynthesized using Achillea biebersteinii flower extract: apoptosis induction in MCF-7 cells via caspase activation and regulation of Bax and Bcl-2 gene expression. Molecules 2015;20(2): 2693-706.##Marslin G, Siram K, Maqbool Q, Selvakesavan RK, Kru-szka D, Kachlicki P, et al. Secondary metabolites in the green synthesis of metallic nanoparticles. Materials (Basel) 2018;11(6):940. ##Inbathamizh L, Ponnu TM, Mary EJ. In vitro evaluation of antioxidant and anticancer potential of Morinda pu-bescens synthesized silver nanoparticles. J Pharmacy Re-search 2013;6(1):32-8.##Ahmed S, Ahmad M, Swami BL, Ikram S. A review on plants extract mediated synthesis of silver nanoparticles for antimicrobial applications: a green expertise. 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Investigation of Durability of SARS-CoV-2-specific IgG and IgM Antibodies in Recovered COVID-19 Patients: A Prospective Study 2 2 Background: Evidence on seroconversion profile of the novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infected patients is limited. We mainly aimed to evaluate seroconversion and persistence of virus-specific antibodies in patients infected by coronavirus disease 2019 (COVID-19). Methods: This prospective study was conducted on 118 patients with COVID-19 presentations admitted to three hospitals in Iran and recovered from the disease, during April and May 2020. Presence of COVID-19 was confirmed by Polymerase Chain Reaction (PCR) testing on nasopharyngeal swabs. Serum samples were collected at different time points, including 0-5, 6-15, 16-25, 26-35, and 36-95 days of clinical symptom onset. For measurement of SARS-CoV-2-specific IgG and IgM antibody titers, Iran's Food and Drug Administration-approved SARS-CoV-2 ELISA kits were used. Results: Serologic assay revealed that 37.3% of patients (n=44) were positive for IgM at 0-5 days interval after clinical symptom onset. This rate was 60.2% (n=71) for IgG. There were increasing IgM and IgG seroconversion rates during first 25 days of clinical symptom onset, but seropositivity started to decrease thereafter, which was more evident for IgM (17.9%) than IgG (58.9%) at the 36-95 days post symptoms appearance. In other words, it was found that 83.6% of IgM-positive and 32.9% of IgG-positive patients in the first month of clinical symptom onset became seronegative in the third month of clinical symptom onset. Conclusion: The findings demonstrated that antibody responses to SARS-CoV-2 infection were developed in recovered COVID-19 patients; however, some of them were seronegative three months after onset of relevant symptoms. Furthermore, the stability of anti-SARS-CoV-2 antibodies could also correct our expectations from COVID-19 vaccination responses. 233 238 Mohammad Zamani Digestive Diseases Research Institute, Shariati Hospital, Tehran University of Medical Sciences Iran Ahmad Ghasemi Healthy Ageing Research Centre, Neyshabur Faculty of Medical Sciences, Neyshabur University Medical Sciences Iran Morteza Shamshirgaran Healthy Ageing Research Centre, Neyshabur Faculty of Medical Sciences, Neyshabur University Medical Sciences Iran Sajjad Ahmadpour Gastroenterology and Hepatology Diseases Research Center, Qom University of Medical Sciences Iran Ahmad Hormati Gastroenterology and Hepatology, Department of Internal Medicine, School of Medicine, Gastrointestinal and Liver Diseases Research Center, Iran University of Medical Sciences Iran Javad Khodadadi Infectious Disease Department, Qom University of Medical Sciences Infectious Disease Department, Qom University of Medical Sciences Iran Mehran Varnasseri Infectious and Tropical Diseases Research Center, Ahvaz Jundishapur University of Medical Sciences Iran Fatemeh Amini Department of Persian Medicine, School of Medicine, Shiraz University of Medical Sciences Department of Persian Medicine, School of Medicine, Shiraz University of Medical Sciences Iran Amaneh Shayanrad Liver and Pancreatobiliary Diseases Research Center, Digestive Diseases Research Institute, Shariati Hospital, Tehran University of Medical Sciences Iran Vahid Younesi Synapse IVD Accelerator Iran Hossein Poustchi Liver and Pancreatobiliary Diseases Research Center, Digestive Diseases Research Institute, Shariati Hospital, Tehran University of Medical Sciences Iran Mahdi Shabani Department of Immunology, School of Medicine, Shahid Beheshti University of Medical Sciences Department of Immunology, School of Medicine, Shahid Beheshti University of Medical Sciences Iran COVID-19SARS-CoV-2SeroconversionSeropositivity 60509.pdf Oude Munnink BB, Worp N, Nieuwenhuijse DF, Sik-kema RS, Haagmans B, Fouchier RA, et al. 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Prioritizing Candidate Genes for Type 2 Diabetes Mellitus using Integrated Network and Pathway Analysis 2 2 Background: Type 2 Diabetes Mellitus (T2DM) has emerged as a major threat to global health that fosters life-threatening clinical complications, taking a huge toll on our society. More than 65 million Indians suffer from T2DM, making it one of the leading causes of death. T2DM and associated complications have to be constantly monitored and managed which reduces the overall quality of life and increases socioeconomic burden. Therefore, it is crucial to develop specific treatment and management strategies. In order to achieve this, it is essential to understand the underlying genetic causes and molecular mechanisms. Methods: Integrated gene network and ontology analyses facilitate prioritization of plausible candidate genes for T2DM and also aid in understanding their mechanistic pathways. In this study, T2DM-associated genes were subjected to sequential interaction network and gene set enrichment analysis. High ranking network clusters were derived and their interrelation with pathways was assessed. Results: About 23 significant candidate genes were prioritized from 615 T2DM-associ-ated genes which were overrepresented in pathways related to insulin resistance, type 2 diabetes, signaling cascades such as insulin receptor signaling pathway, PI3K signaling, IGFR signaling pathway, ERBB signaling pathway, MAPK signaling pathway and their regulatory mechanisms. Conclusion: Of these, two tyrosine kinase receptor genes-EGFR and IGF1R were identified as common nodes and can be considered to be significant candidate genes in T2DM. <div> <div> <div> </div> <div> <div> <div> </div> <div>     </div> </div> </div> </div> </div> 239 246 Tejaswini Prakash Genetics and Genomics Lab, Department of Studies in Genetics and Genomics, University of Mysore, Manasagangothri, Mysuru – 570 006 Genetics and Genomics Lab, Department of Studies in Genetics and Genomics, University of Mysore, Manasagangothri, Mysuru – 570 006 India Nallur B Ramachandra Genetics and Genomics Lab, Department of Studies in Genetics and Genomics, University of Mysore, Manasagangothri, Mysuru – 570 006 Genetics and Genomics Lab, Department of Studies in Genetics and Genomics, University of Mysore, Manasagangothri, Mysuru – 570 006 India Gene ontologyHub genes identificationIn silico analysisText miningType 2 diabetes mellitus 60510.pdf World Health Organization. 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An in silico Design, Expression and Purification of a Chimeric Protein as an Immunogen Candidate Consisting of IpaD, StxB, and TolC Proteins from Shigella spp. 2 2 Background: Shigella spp. is the cause of dysentery and is widespread worldwide. On the other hand, antibiotic resistance is increasing in this bacterium. Bioinformatics is a new approach to vaccine and drug design involving the selection of appropriate antigens. This study aimed to design a chimeric protein consisting of IpaD, StxB, and TolC proteins from Shigella through a bioinformatics approach as an immunogen candidate. Methods: The sequences of ipaD, stxB, and tolC genes were obtained. Additionally, the immunogenic regions of the associated protein, physicochemical characteristics, protein structures, B and T cells epitopes, and molecular docking were determined using in silico servers. Besides, the chimeric gene was synthesized following sequence optimization by utilizing the codon usage of Escherichia coli (E. coli). The expression of the recombinant protein was confirmed via SDS-PAGE and Western blot technique. Results: The residues 41-160 of IpaD, 21-89 of StxB, and 40-335 of TolC were selected. According to half-life, instability, and buried indices, IpaD-StxB-TolC was selected as the best arrangement. The Ramachandran plot showed that 97.077% of the amino acids were in the favored area. Linear and conformational epitopes were also present throughout the chimeric protein sequence. Moreover, the C-ImmSim server indicated that IgG and IgM titers could reach desirable values by the third injection. Furthermore, the stability of the mRNA-optimized gene was enhanced, increasing the Codon Adaptive Index (CAI) to 0.9. Finally, the chimeric gene was transferred to E. coli BL21, and the expression of the 60.6 kDa recombinant protein was confirmed.  Conclusion: The results indicated that the recombinant protein could act as a proper immunogen candidate against Shigella spp. 247 258 Javad Fathi Department of Bacteriology and Virology, Faculty of Medicine, Shiraz University of Medical SciencesStudent Research Committee, Faculty of Medicine, Shiraz University of Medical Sciences Department of Bacteriology and Virology, Faculty of Medicine, Shiraz University of Medical SciencesStudent Research Committee, Faculty of Medicine, Shiraz University of Medical Sciences IranIran Shahram Nazarian Emad Kordbacheh Department of Biological Sciences, Faculty of Science, Imam Hossein University Department of Biological Sciences, Faculty of Science, Imam Hossein University Iran Nahal Hadi Department of Bacteriology and Virology, Faculty of Medicine, Shiraz University of Medical Sciences Department of Bacteriology and Virology, Faculty of Medicine, Shiraz University of Medical Sciences Iran Computer simulationDysenteryRecombinant fusion proteinsShigella sppShigella vaccine candidate 60511.pdf Petri WA, Miller M, Binder HJ, Levine MM, Dillingham R, Guerrant RL. 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Impact of Single Nucleotide Polymorphism in the ANKRD55 Gene on Occurrence and Clinical Characteristics of Rheumatoid Arthritis 2 2 Background: Rheumatoid Arthritis (RA) has multifactorial etiology and numerous genetic and environmental factors have been related to an increased risk of RA. Recently, Genome-Wide Association Studies (GWAS) suggested a large number of Single Nucleotide Polymorphisms (SNPs) loci affecting the susceptibility to RA. One of these loci is rs6859219 (C>A), a functional polymorphism in the ANKRD55 gene which was associated with the expression of ANKRD55 and IL6ST. In the current study, we evaluated the possible association between rs6859219 (intronic variant) in the ANKRD55 gene with RA risk in the Iranian population. Methods: A case-control study using 118 RA patients and 115 healthy counterparts was undertaken in order to determine rs6859219 genotypes using real‑time polymerase chain reaction High‑Resolution Melting (HRM) method. Results: There was a significant difference in the genotype and allele frequencies of rs6859219 between patients and controls (p<0.001). Logistic regression analysis demonstrates that CC genotype and C allele increased the risk of RA (OR for CC genotype= 7.12; 95%CI [3.51-15.05]/ OR for C allele=4.16; 95%CI [2.78-6.28]). Furthermore, regarding the dominant and recessive model of inheritance, RA patients indicated obvious association of the rs6859219 variant compared to healthy controls (p<0.001). Moreover, in the patient group, there was a significant correlation between C-Reactive Protein (CRP) concentration with rs6859219 polymorphism (p<0.001). Conclusion: Our findings propose a substantial correlation between rs6859219 polymorphism and RA risk and clinical characteristics of this disease in the Iranian population. 259 263 Rasoul Salehi Pediatric Inherited Diseases Research Center, Research Institute for Primordial Prevention of Non-Communicable Disease and Department of Genetics and Molecular Biology, School of Medicine, Isfahan University of Medical Sciences Iran Mina Motaghi Division of Rheumatology, Department of Internal Medicine, School of Medicine, Isfahan University of Medical Sciences Division of Rheumatology, Department of Internal Medicine, School of Medicine, Isfahan University of Medical Sciences Iran Amirhossein Salehi Division of Rheumatology, Department of Internal Medicine, School of Medicine, Isfahan University of Medical Sciences Division of Rheumatology, Department of Internal Medicine, School of Medicine, Isfahan University of Medical Sciences Iran Hadi Karimzadeh Division of Rheumatology, Department of Internal Medicine, School of Medicine, Isfahan University of Medical Sciences Division of Rheumatology, Department of Internal Medicine, School of Medicine, Isfahan University of Medical Sciences Iran Bahram Pakzad Division of Rheumatology, Department of Internal Medicine, School of Medicine, Isfahan University of Medical Sciences Division of Rheumatology, Department of Internal Medicine, School of Medicine, Isfahan University of Medical Sciences Iran Autoimmune diseaseIranRheumatoid arthritisSingle nucleotide polymorphisms 60512.pdf Deane KD, Demoruelle MK, Kelmenson LB, Kuhn KA, Norris JM, Holers VM. 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