Basic Science and Clinical Studies on Non-COVID-19 Topics, of Coronavirus Victims 2 2 Despite advancements of societies in terms of health, epidemics do happen; the latest is the outbreak of coronavirus. Re-assortment of viruses is a biological phenomenon that creates new subtypes of viruses, indicating that pandemics are unavoidable 1. The SARS-CoV-2 crisis has so far affected all countries of the world, which has been more pronounced in aspects related to health and economy. One area in which COVID-19 has had a significant impact is research 2. The fact that a pandemic of a virus has caused and continues to cause deaths worldwide and dire consequences for the economies of nations are of main reasons for the focus of research in this field. Currently, people and the media are paying the most attention to the disease among other health-related issues, and governments are asking researchers to find solutions to the problem has negatively affected all of society. As a result, research centers' budgets are prioritized with issues related to COVID-19, and the majority of researchers have focused on the disease 2-4. In this situation, are basic science research and diseases other than COVID-19 given necessitous attention 5-7? The large number of researchers focusing on the field and the rapidity of the publication of research, despite its many advantages, has several significant disadvantages that affect both COVID-19 related and other research. Rapid and immature conclusions from some trials and their use in the clinic, despite the low quality of several studies and small size of study samples-compared to the COVID-19 spectrum-along with disruption of the peer review process even in reputable journals are of the significant drawbacks affecting COVID-19-related research 3,8. The negative impact of this pandemic on clinical trials on topics other than COVID-19 is worrying. Due to the transmission problem required to perform many trials, performing such studies is problematic in terms of implementation and practice, but the main problem goes back to before this stage. Governments want to resolve the coronavirus crisis, people and the media are following the issue of COVID-19, COVID-19 is a priority for research center budgets, and journals publish research on the field faster; as a result, individual interests such as advancement in academic status and financial profits in this field are better secured 2-8. Delays in progress in basic science and clinical research other than COVID-19 due to budget cuts and reduced researcher focus will have long-term adverse effects 9. Doing basic science and free studies-that is, studies for which there are no questions or requests-often leads to other questions and sometimes to answers to which we have never asked a question. The results of these studies are knowledge and understanding and are necessary for further studies. These studies are the entrance to a bridge built from the laboratory to the clinic to the community 9. Science works this way, from free studies to patient treatment, and the result of a reduction in funding and a reduction in basic science and clinical studies other than COVID-19 will be nothing but a delay in the advancement of science in general. 1 1 Shahin Akhondzadeh Ahmad Shamabadi Editorial 30441.pdf Potter CW. A history of influenza. J Appl Microbiol 2001;91(4):572-9. ##Donthu N, Gustafsson A. Effects of COVID-19 on business and research. J Bus Res 2020;117:284-9.##Harper L, Kalfa N, Beckers GMA, Kaefer M, Nieuwhof-Leppink AJ, Fossum M, et al. The impact of COVID-19 on research. J Pediatr Urol 2020.##Webster P. How is biomedical research funding faring during the COVID-19 lockdown? Nat Med 2020.##Noorbala AA, Akhondzadeh S, Davari-Ashtiani R, Amini-Nooshabadi H. Piracetam in the treatment of schizophrenia: implications for the glutamate hypothesis of schizophrenia. J Clin Pharm Ther 1999;24(5):369-74.##Akhondzadeh S. The 5-HT hypothesis of schizophrenia. IDrugs 2001;4(3):295-300. ##Kashani L, Omidvar T, Farazmand B, Modabbernia A, Ramzanzadeh F, Tehraninejad ES, et al. Does pioglitazone improve depression through insulin-sensitization? Results of a randomized double-blind metformin-controlled trial in patients with polycystic ovarian syndrome and comorbid depression. Psychoneuroendocrinology 2013;38(6):767-76.##Rabin R. The pandemic claims new victims: Prestigious Medical Journals. The New York Times June 15, 2020.##Schor NF. Why our patients (and we) need basic science research. Neurology 2013;80(22):2070-5.##

Silencing SALL-4 Gene by Transfecting Small Interfering RNA with Targeted Aminoglycoside-Carboxyalkyl Polyethylenimine Nano-Polyplexes Reduced Migration of MCF-7 Breast Cancer Cells 2 2 Background: The application of non-viral systems for delivering genes to cells is becoming a very interesting issue, especially in the treatment of neoplasms such as Breast Cancer (BC). Polymer-based non-viral systems are safe and feasible gene carriers to be used in targeted cancer therapy. SALL4 gene encodes a transcription factor and is overexpressed in some cancers. Methods: In this study, carboxyalkylated-PEI25 (25 kDa) was used to deliver plasmids expressing SALL4-siRNA into MCF-7 cells. DLS and AFM were applied to determine the size of nanoparticles. The MTT method was used to assess cytotoxicity, and the efficiency of transfection was confirmed both qualitatively and quantitatively. Finally, the effect of silencing SALL4 was investigated on the migration of MCF7 cells using the scratch test. Results: The results showed that transferring the SALL4-siRNA using PEI25G10C50 reduced the expression of the corresponding transcription factor by 14 folds which attenuated the migration of MCF-7 cells by 58%. Conclusion: In conclusion, PEI25G10C50 can serve as an effective gene delivery system for treating BC by targeting SALL-4. 2 8 Somaye Noruzi Department of Advanced Sciences and Technologies, Faculty of Medicine, North Khorasan University of Medical SciencesStudent Research Committee, Faculty of Medicine, North Khorasan University of Medical Sciences Department of Advanced Sciences and Technologies, Faculty of Medicine, North Khorasan University of Medical SciencesStudent Research Committee, Faculty of Medicine, North Khorasan University of Medical Sciences IranIran Mehran Vatanchian Department of Anatomical Sciences, Faculty of Medicine, North Khorasan University of Medical Sciences Department of Anatomical Sciences, Faculty of Medicine, North Khorasan University of Medical Sciences Iran Amir Azimian Department of Pathobiology and Laboratory Sciences, Faculty of Medicine, North Khorasan University of Medical Sciences Department of Pathobiology and Laboratory Sciences, Faculty of Medicine, North Khorasan University of Medical Sciences Iran Arash Niroomand Department of Advanced Sciences and Technologies, Faculty of Medicine, North Khorasan University of Medical SciencesStudent Research Committee, Faculty of Medicine, North Khorasan University of Medical Sciences Department of Advanced Sciences and Technologies, Faculty of Medicine, North Khorasan University of Medical SciencesStudent Research Committee, Faculty of Medicine, North Khorasan University of Medical Sciences IranIran Reza Salarinia Fatemeh Oroojalian Breast neoplasmsGene transfer techniquesMCF7 cellsTranscription factors 30437.pdf Bray F, Ferlay J, Soerjomataram I, Siegel RL, Torre LA, Jemal A. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin 2018;68(6):394-424. ##Gradishar WJ, Anderson BO, Balassanian R, Blair SL, Burstein HJ, Cyr A, et al. Breast cancer Version 2.2015. J Natl Compr Canc Netw 2015;13(4):448-75.##Mollen EWJ, Ient J, Tjan-Heijnen VCG, Boersma LJ, Miele L, Smidt ML, et al. Moving breast cancer therapy up a notch. Front Oncol 2018;8:518.##Xue HY, Wong H. Targeting megalin to enhance delivery of anti-clusterin small-interfering RNA nanomedicine to chemo-treated breast cancer. Eur J Pharm Biopharm 2012;81(1):24-32. ##Chen LP, Zhang NN, Ren XQ, He J, Li YJM. miR-103/miR-195/miR-15b regulate SALL4 and inhibit proliferation and migration in glioma. Molecules 2018;23(11):2938. ##Zhang L, Xu Z, Xu X, Zhang B, Wu H, Wang M, et al. SALL4, a novel marker for human gastric carcinogenesis and metastasis. Oncogene 2014;33(48):5491-500.##Yang JJBR. SALL4 as a transcriptional and epigenetic regulator in normal and leukemic hematopoiesis.Biomark Res 2018;6(1):1.##Yue X, Xiao L, Yang Y, Liu W, Zhang K, Shi G, et al. High cytoplasmic expression of SALL4 predicts a malignant phenotype and poor prognosis of breast invasive ductal carcinoma. Neoplasma 2015;62(6):980-8. ##Forghanifard MM, Moghbeli M, Raeisossadati R, Tavassoli A, Mallak AJ, Boroumand-Noughabi S, et al. Role of SALL4 in the progression and metastasis of colorectal cancer. J Biomed Sci 2013;20(1):6.##Zhang X, Yuan X, Zhu W, Qian H, Xu WJCl. SALL4: an emerging cancer biomarker and target. Cancer Lett 2015;357(1):55-62. ##Zhang X, Zhang P, Shao M, Zang X, Zhang J, Mao F, et al. SALL4 activates TGF-β/SMAD signaling pathway to induce EMT and promote gastric cancer metastasis. Cancer Manag Res 2018;10:4459-70. ##Oh YK, Park TG. siRNA delivery systems for cancer treatment. Adv Drug Deliv Rev 2009;61(10):850-62. ##Lin G, Zhang H, Huang LJMp. Smart polymeric nanoparticles for cancer gene delivery. Mol Pharm 2015;12(2):314-21. ##Rashidi A, Omidi M, Choolaei M, Nazarzadeh M, Yadegari A, Haghierosadat F, et al. Electromechanical properties of vertically aligned carbon nanotube. Advanced Materials Research 2013;705:332-36.##Oroojalian F, Charbgoo F, Hashemi M, Amani A, Yazdian-Robati R, Mokhtarzadeh A, et al. Recent advances in nanotechnology-based drug delivery systems for the kidney. J Controlled Release 2020;321:442-62. ##Oroojalian F, Rezayan AH, Shier WT, Abnous K, Ramezani M. Megalin-targeted enhanced transfection efficiency in cultured human HK-2 renal tubular proximal cells using aminoglycoside-carboxyalkyl-polyethylenimine-containing nanoplexes. Int J Pharm 2017;523(1):102-20. ##Oroojalian F, Jahanafrooz Z, Chogan F, Rezayan AH, Malekzade E, Rezaei SJT, et al. Synthesis and evaluation of injectable thermosensitive penta‐block copolymer hydrogel (PNIPAAm‐PCL‐PEG‐PCL‐PNIPAAm) and star‐shaped poly (CL─ CO─ LA)‐b‐PEG for wound healing applications. J CellBiochem 2019;120(10):17194-207.##Pishavar E, Oroojalian F, Ramezani M, Hashemi M. Cholesterol‐conjugated PEGylated PAMAM as an efficient nanocarrier for plasmid encoding interleukin‐12 immunogene delivery towards colon cancer cells. Biotechnol Prog 2019;36(3):e2952.##Rödl W, Schaffert D, Wagner E, Ogris M. Synthesis of polyethylenimine-based nanocarriers for systemic tumor targeting of nucleic acids. Methods Mol Biol 2013;948:105-20. ##Schmid SL. Reciprocal regulation of signaling and endocytosis: Implications for the evolving cancer cell. J Cell Biol 2017;216(9):2623-32. ##Pandey AP, Sawant KK. Polyethylenimine: A versatile, multifunctional non-viral vector for nucleic acid delivery. Materials Science and Engineering: C 2016;68:904-18. ##Mahadevappa R, Nielsen R, Christensen EI, Birn H. Megalin in acute kidney injury: foe and friend. Am J Physiol Renal Physiol 2014;306(2):F147-F54. ##Patnaik S, Gupta KC. Novel polyethylenimine-derived nanoparticles for in vivo gene delivery. Expert Opin Drug Deliv 2013;10(2):215-28. ##Nayerossadat P, Kichler A. Recent developments in nucleic acid delivery with polyethylenimines. Adv Genet 2014;88:263-88. ##Hesari A, Anoshiravani AA, Talebi S, Noruzi S, Mohammadi R, Salarinia R, et al. Knockdown of sal-like 4 expression by small interfering RNA induces apoptosis in breast cancer cells. J Cell Biochem 2019;120(6):9392-9. ##McCrudden CM, McCarthy HO. Current status of gene therapy for breast cancer: progress and challenges. Appl Clin Genet 2014;7:209.##Nayerossadat N, Maedeh T, Abas ali P. Viral and nonviral delivery systems for gene delivery. Adv Biomed Res 2012;1:27. ##Zhang J, Li X, Huang L. Non-viral nanocarriers for siRNA delivery in breast cancer. J Control Release 2014;190:440-50. ##Niebel Y, Buschmann MD, Lavertu M, De Crescenzo G. Combined analysis of polycation/ODN polyplexes by analytical ultracentrifugation and dynamic light scattering reveals their size, refractive index increment, stoichiometry, porosity, and molecular weight. Biomacromolecules 2014;15(3):940-7. ##Choosakoonkriang S, Lobo BA, Koe GS, Koe JG, Middaugh CR. Biophysical characterization of PEI/DNA complexes. J Pharm Sci 2003;92(8):1710-22. ##Hickey JW, Santos JL, Williford JM, Mao HQ. Control of polymeric nanoparticle size to improve therapeutic delivery. J Control Release 2015;219:536-47. ##Itou J, Tanaka S, Li W, Iida A, Sehara-Fujisawa A, Sato F, et al. The Sal-like 4-integrin α6β1 network promotes cell migration for metastasis via activation of focal adhesion dynamics in basal-like breast cancer cells. Biochimica et Biophysica acta. Molecular Cell Research 2017;1864(1):76-88. ##Wang M, Qiu R, Gong Z, Zhao X, Wang T, Zhou L, et al. miR‐188‐5p emerges as an oncomiRNA to promote gastric cancer cell proliferation and migration via upregulation of SALL4. J Cell Biochem 2019 Sep;120(9):15027-37.##Jiang X, Wang Z. miR-16 targets SALL4 to repress the proliferation and migration of gastric cancer. Oncol Lett 2018;16(3):3005-12.##

Evaluation of Metastasis Suppressor Genes Expression and In Vitro Anti-Cancer Effects of Zinc Oxide Nanoparticles in Human Breast Cancer Cell Lines MCF-7 and T47D 2 2 Background: Metallic nanoparticles are useful materials to be applied in biomedical research. In this study, the possible apoptotic and anti-metastatic activity of Zinc Oxide Nanoparticles (ZnONPs) was assessed in breast cancer cells. Methods: First, in vitro cell viability was investigated by MTT assay in two human breast cancer cells (MCF-7 and T47D) and normal Human Embryonic Kidney (HEK293) cells at 37°C overnight. Apoptosis induced by ZnONPs was evaluated by annexin V/PI staining, cell cycle analysis and caspase assay in cancerous cells. Moreover, quantitative real-time PCR was employed for the detection of two metastasis suppressor genes (KAI-1 and NM23) expression in cancerous cells. Results: Data demonstrated that ZnONPs exert a dose-dependent inhibitory effect on the viability of T47D and MCF-7 cells, while no cytotoxic effect was observed on normal HEK293 cells. The mRNA expression levels of KAI-1 and non-metastatic protein (NM23) genes were up-regulated in ZnONP-exposed cancerous cells. ZnONPs were also found to enhance the apoptosis properties of cells by annexin V/PI staining, and caspase assay in cancerous cells. Furthermore, ZnONPs can increase sub-G1 population as compared to negative control. Conclusion: Our findings showed that ZnONPs induce apoptotic activity and can modulate metastasis by up-regulating of KAI-1 and NM23 gene expression in two breast cancer (MCF-7 and T47D) cells. 9 14 Seyed Ataollah Sadat Shandiz Department of Biology, Central Tehran Branch, Islamic Azad University Department of Biology, Central Tehran Branch, Islamic Azad University Iran Faryad Sharifian Department of Genetics and Biotechnology, Faculty of Biological Sciences, Varamin-Pishva Branch, Islamic Azad University Department of Genetics and Biotechnology, Faculty of Biological Sciences, Varamin-Pishva Branch, Islamic Azad University Iran Sorayya Behboodi Department of Biology, Tehran Shargh (East), Payam Noor University Department of Biology, Tehran Shargh (East), Payam Noor University Iran Fatemeh Ghodratpour Department of Genetics and Biotechnology, Faculty of Biological Sciences, Varamin-Pishva Branch, Islamic Azad University Department of Genetics and Biotechnology, Faculty of Biological Sciences, Varamin-Pishva Branch, Islamic Azad University Iran Fahimeh Baghbani-Arani ApoptosisBreast neoplasmsCell cycleZinc oxide 40442.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):E359-86.##Pedersen RN, Bhaskaran K, Heide Jorgensen U, Norgaard M, Christiansen PM¬, Kroman N, et al. Breast cancer recurrence after reoperation for surgical bleeding. Br J Surg 2017;104(12):1665-74.##Lee J, Chatterjee DK, Lee MH¬, Krishnan S. Gold nanoparticles in breast cancer treatment: Promise and potential pitfalls. Cancer Lett 2014;347(1):46-53. ##Rasmussen JW, Martinez E, Louka P, Wingett DG. Zinc oxide nanoparticles for selective destruction of tumor cells and potential for drug delivery applications. Expert Opin Drug Deliv 2010;7(9):1063-77. ##Chandrasekaran M, Pandurangan M. In vitro selective anti-proliferative effect of zinc oxide nanoparticles against co-cultured C2C12 myoblastoma cancer and 3T3-L1 normal cells. Biol Trace Elem Res 2016;172(1):148-54. ##Malhotra SPK, Mandal TK. Biomedical applications of Zinc Oxide nanomaterials in cancer treatment: a review. SCIREA J Chem 2016;1(2):67-89. ##Yakimova R, Selegard L, Khranovskyy V, Pearce R, Spetz AL, Uvdal K. ZnO materials and surface tailoring for biosensing. Front Biosci (Elite Ed) 2012;4:254-78. ##Akhtar MJ, Ahamed M, Kumar S, Khan MM, Ahmad J, Alrokayan SA. Zinc oxide nanoparticles selectively induce apoptosis in human cancer cells through reactive oxygen species. Int J Nanomed 2012;7:845–57.##Wahab R, Siddiqui MA, Saquib Q, Dwivedi S, Ahmad J, Musarrat J, et al. ZnO nanoparticles induced oxidative stress and apoptosis in HepG2 and MCF-7 cancer cells and their antibacterial activity. Coll Surfaces B: Biointerfaces 2014;117(1):267-76.##Buxton ILO, Yokdang N. Extracellular Nm23 Signaling in breast cancer: incommodus verum. Cancers (Basel) 2011;3(3):2844-57. ##Howlett AR, Petersen OW, Steeg PS, Bissel MJ. A novel function for the nm23-Hl gene: overexpression in human breast carcinoma cells leads to the formation of basement membrane and growth arrest. J Natl Cancer Inst 1994;86(24):1838-44.##Phillips KK, White AE, Hicks DJ, Welch DR, Barrett JC, Wei LL, et al. Correlation between reduction of metastasis in the MDA-MB-435 model system and increased expression of the Kai-1 protein. Mol Carcinog 1998;21(2):111-20.##Wei LL, Yang X, Phillips KK, Weissman BE, Welch DR. Analysis of KAI-1 mRNA expression in human breast cancer cell lines. Proc Am Assoc Cancer Res 1996;37:529.##Hurst DR, Welch DR. Metastasis suppressor genes: at the interface between the environment and tumor cell growth. Int Rev Cell Mol Biol 2011;286:107-80. ##Liu FS, Dong JT, Chen JT, Hsieh YT, Ho ES, Hung MJ. Frequent down-regulation and lack of mutation of the KAI1 metastasis suppressor gene in epithelial ovarian carcinoma. Gynecol Oncol 2000;78(1):10-15. ##Moshfegh A, Salehzadeh A, Shandiz SAS, Shafaghi M, Naeemi AS, Salehi S. Phytochemical analysis, antioxidant, anticancer and antibacterial properties of the Caspian sea red Macroalgae, Laurencia caspica. Iran J Sci Technol Trans Sci 2019;43(1):49-56.##Foldbjerg R, Olesen P, Hougaard M, Dang DA, Hoffmann HJ, Autrup H. PVP-coated silver nanoparticles and silver ions induce reactive oxygen species, apoptosis and necrosis in THP-1 monocytes. Toxicol Lett 2009;190(2):156-62.##Arora S, Jain J, Rajwade JM, Paknikar KM. Cellular responses induced by silver nanoparticles: In vitro studies. Toxicol Lett 2008;179(2):93-100. ##Asare N, Instanes C, Sandberg WJ, Refsnes M, Schwarze P, Kruszewski M, et al. Cytotoxic and genotoxic effects of silver nanoparticles in testicular cells. Toxicology 2012;291(1-3):65-72. ##You J, Chang R, Liu B, Zu L, Zhou Q. Nm23-H1 was involved in regulation of KAI1 expression in high-metastatic lung cancer cells L 9981. J Thorac Dis 2016;8(6):1217-26. ##Yoshida BA, Sokoloff MM, Welch DR, Rinker-Shaeffer CW. Metastasis-suppressor genes: A review and perspective on an emerging field. J Natl Cancer Inst 2000;92(21):1717-30. ##Bozdogan O, Yulug IG, Vargel I, Cavusoglu T, Karabulut AA, Karahan G, et al. Differential expression patterns of metastasis suppressor proteins in basal cell carcinoma. Int J Dermatol 2015;54(8):905-15.##Stark AM, Tongers K, Maass N, Mehdorn HM, Held-Feindt J. Reduced metastasis-suppressor gene mRNA-expression in breast cancer brain metastases, J Cancer Res Clin Oncol 2005;131(3):191-8.##Sheikh BY, Sarker MMR, Kamarudin MNA, Mohan G. Antiproliferative and apoptosis inducing effects of citral via p53 and ROS induced mitochondrial-mediated apoptosis in human colorectal HCT116 and HT29 cell lines. Biomed Pharma 2017;96:834-46. ##Asharani PV, Hande MP, Valiyaveettil S. Anti-proliferative activity of silver nanoparticles. BMC Cell Biol 2009;10:65.##Turk B, Stoka V. Protease signalling in cell death: caspases versus cysteine cathepsins. FEBS Letters 2007;581(15):2761-7. ##Puiso J, Jonkuviene D, Macioniene I, Salomskiene J, Jasutiene I, Kondrotas R. Biosynthesis of silver nanoparticles using lingonberry and cranberry juices and their antimicrobial activity, Colloids Surfaces B Biointerfaces 2014;121:214-21. ##Krishna PG, Ananthaswamy PP, Yadavalli T, Mutta NB, Sannaiah A, Shivanna Y. ZnO nanopellets have selective anticancer activity. Mater Sci Eng C Mater Biol Appl 2016;62:919-26. ##Pandurangan M, Enkhtaivan G, Kim DH. Anticancer studies of synthesized ZnO nanoparticles against human cervical carcinoma cells. Photochem J Photobiol B Biol 2016;58:206-11.##Mariappan P, Krishnamoorthy K, Kadarkaraithangam J, Govindasamy M. Selective toxicity of ZnO nanoparticles toward gram-positive bacteria and cancer cells by apoptosis through lipid peroxidation. Nanomedicine 2011;7(2):184-92. ##Boroumand Moghaddam A, Moniri M, Azizi S, Abdul Rahim R, Ariff AB, Navaderi M, et al. Eco-friendly formulated zinc oxide nanoparticles: induction of cell cycle arrest and apoptosis in the MCF-7 cancer cell line. Genes (Basel) 2017;8(10):281. ##

Integrated Network and Gene Ontology Analysis Identifies Key Genes and Pathways for Coronary Artery Diseases 2 2 Background: The prevalence of Coronary Artery Disease (CAD) in developing countries is on the rise, owing to rapidly changing lifestyle. Therefore, it is imperative that the underlying genetic and molecular mechanisms be understood to develop specific treatment strategies. Comprehensive disease network and Gene Ontology (GO) studies aid in prioritizing potential candidate genes for CAD and also give insights into gene function by establishing gene and disease pathway relationships. Methods: In the present study, CAD-associated genes were collated from different data sources and protein-protein interaction network was constructed using STRING. Highly interconnected network clusters were inferred and GO analysis was performed. Results: Interrelation between genes and pathways were analyzed on ClueGO and 38 candidates were identified from 1475 CAD-associated genes, which were significantly enriched in CAD-related pathways such as metabolism and regulation of lipid molecules, platelet activation, macrophage derived foam cell differentiation, and blood coagulation and fibrin clot formation. Discussion: Integrated network and ontology analysis enables biomarker prioritization for common complex diseases such as CAD. Experimental validation and future studies on the prioritized genes may reveal valuable insights into CAD development mechanism and targeted treatment strategies. 15 23 Tejaswini Prakash Genetics and Genomics Lab, Department of Studies in Genetics and Genomics, University of Mysore Genetics and Genomics Lab, Department of Studies in Genetics and Genomics, University of Mysore India Nallur B Ramachandra Coronary artery diseaseFibrinGene ontologyPathway enrichmentProtein interaction maps 40443.pdf Lightbody G, Haberland V, Browne F, Taggart L, Zheng H, Parkes E, et al. Review of applications of high-throughput sequencing in personalized medicine: barriers and facilitators of future progress in research and clinical application. Brief Bioinform 2019; 20(5):1795-811. ##Hofker MH, Fu J, Wijmenga C. The genome revolution and its role in understanding complex diseases. Biochim Biophys Acta 2014;1842(10):1889-95. ##Mc Namara K, Alzubaidi H, Jackson JK. Cardiovascular disease as a leading cause of death: how are pharmacists getting involved? Integr Pharm Res Pract 2019;4(8):1-11.##Ghatge M, Nair J, Sharma A, Vangala RK. Integrative gene ontology and network analysis of coronary artery disease associated genes suggests potential role of ErbB pathway gene EGFR. Mol Med Rep 2018;17(3):4253-64. ##Hajar R. Risk factors for coronary artery disease: Historical perspectives. Heart Views 2017;18(3):109-14.##CARDIoGRAMplusC4D Consortium; Deloukas P, Kanoni S, Willenborg C, Farrall M, Assimes TL, Thompson JR, et al: Large‑scale association analysis identifies new risk loci for coronary artery disease. Nature Genetics 2013;45:25-33.##Mehta NN. Large‑scale association analysis identifies 13 new susceptibility loci for coronary artery disease. Circ Cardiovasc Genet 2011;4(3):327-9.##Arvind P, Jayashree S, Jambunathan S, Nair J, Kakkar VV. Understanding gene expression in coronary artery disease through global profiling, network analysis and independent validation of key candidate genes. J Genet 2015;94(4):601-10.##Doncheva NT, Kacprowski T, Albrecht M. Recent approaches to the prioritization of candidate disease genes. Wiley Interdiscip Rev Syst Biol Med 2012;4(5):429‐42.##Tan X, Zhang X, Pan L, Tian X, Dong P. Identification of key pathways and genes in advanced coronary atherosclerosis using bioinformatics analysis. Biomed Res Int 2017;2017:4323496. ##Kaimal V, Sardana D, Bardes EE, Gudivada RC, Chen J, Jegga AG. Integrative systems biology approaches to identify and prioritize disease and drug candidate genes. Methods Mol Biol 2011;700:241-59. ##Chen BS, Yang SK, Lan CY, Chuang YJ. A systems biology approach to construct the gene regulatory network of systemic inflammation via microarray and databases mining. BMC Med Genomics 2008;1:46.##Nair J, Ghatge M, Kakkar VV, Shanker J. Network analysis of inflammatory genes and their transcriptional regulators in coronary artery disease. PLoS One 2014;9(4):e94328.##Hofree M, Shen JP, Carter H, Ideker T. Network‑based stratification of tumor mutations. Nat Methods 2013;10(11):1108-15.##Makinen VP, Civelek M, Meng Q, Zhang B, Zhu J, Levian C, et al. Integrative genomics reveals novel molecular pathways and gene networks for coronary artery disease. PLoS Genet 2014;10(7):e1004502. ##Zhao Y, Chen J, Freudenberg JM, Meng Q, Rajpal DK, Yang X. Network‑based identification and prioritization of key regulators of coronary artery disease loci. Arterioscler Thromb Vasc Biol 2016;36(5):928-41. ##Sarajlic A, Janjić V, Stojković N, Radak D, Przulj N. Network topology reveals key cardiovascular disease genes. PLoS One 2013;8(8):e71537.##Kashyap S, Kumar S, Agarwal V, Misra DP, Phadke SR, Kapoor A. Protein protein interaction network analysis of differentially expressed genes to understand involved biological processes in coronary artery disease and its different severity. Gene Reports 2018;12:50-60.##Miao L, Yin RX, Huang F, Yang S, Chen WX, Wu JZ. Integrated analysis of gene expression changes associated with coronary artery disease. Lipids Health Disease 2019;18(1):92.##Zhang L, Li X, Tai J, Li W, Chen L. Predicting candidate genes based on combined network topological features: a case study in coronary artery disease. PLoS One 2012;7(6):e39542.##Alexandar V, Nayar PG, Murugesan R, Beaulah M, Darshana P, Ahmed SSSJ. CardioGenBase: A literature based multi-omics database for major cardiovascular diseases. PLoS One 2015;10(12):e0143188. ##Liu Y, Liang Y, Wishart DS. PolySearch 2.0: A significantly improved text-mining system for discovering associations between human diseases, genes, drugs, metabolites, toxins, and more. Nucleic Acids Res 2015;43(W1):W535-42. ##Wang J, Duncan D, Shi Z, Zhang B. WEB-based GEne SeT AnaLysis Toolkit (WebGestalt): update 2013. Nucleic Acids Res 2013;41(Web Server issue):W77-83. ##Szklarczyk D, Gable AL, Lyon D, Junge A, Wyder S, Huerta-Cepas J et al. STRING v11: protein-protein association networks with increased coverage, supporting functional discovery in genome-wide experimental datasets. Nucleic Acids Res 2019;47(D1):D607-D613.##Shannon P, Markiel A, Ozier O, Baliga NS, Wang AT, Ramage D, et al. Cytoscape: a software environment for integrated models of biomolecular interaction networks. Genome Res 2003;13(11):2498‐504.##Zhang YM, Meng LB, Yu SJ, Ma DX. Identification of potential crucial genes in monocytes for atherosclerosis using bioinformatics analysis. J Int Med Res 2020;48(4): 0300060520909277.##Barrett TJ. Macrophages in atherosclerosis regression. Arteriosclerosis, Thrombosis, Vascular Biology 2020;40(1):20-33.##Chistiakov DA, Sobenin IA, Orekhov AN. Vascular extracellular matrix in atherosclerosis. Cardiol Rev 2013;21:270-88.##d'Uscio LV, He T, Katusic ZS. Expression and processing of amyloid precursor protein in vascular endothelium. Physiology (Bethesda) 2017;32(1):20-32.##Tibolla G, Norata GD, Meda C, Arnaboldi L, Uboldi P, Piazza F, et al. Increased atherosclerosis and vascular inflammation in APP transgenic mice with apolipoprotein E deficiency. Atherosclerosis 2010;210(1):78-87.##Austin SA, Combs CK. Amyloid precursor protein mediates monocyte adhesion in AD tissue and apoE(-)/(-) mice. Neurobiology Aging 2010;31(11):1854-66.##Van der Donckt C, Van Herck JL, Schrijvers DM, Vanhoutte G, Verhoye M, Blockx I, et al. Elastin fragmentation in atherosclerotic mice leads to intraplaque neovascularization, plaque rupture, myocardial infarction, stroke, and sudden death. Eur Heart J 2015;36(17):1049-58. ##Kay AM, Simpson CL, Stewart JA Jr. The role of AGE/RAGE signaling in diabetes-mediated vascular calcification. J Diabetes Res 2016;2016(4):1-8.##Liu W, Zhang Y, Yu CM, Ji QW, Cai M, Zhao YX, et al. Current understanding of coronary artery calcification. J Geriatr Cardiol 2015;12(6):668-75.##Senatus LM, Schmidt AM. The AGE-RAGE axis: implications for age-associated arterial diseases. Front Genet 2017;8:187. ##Charchar FJ, Bloomer LD, Barnes TA, Cowley MJ, Nelson CP, Wang Y, et al. Inheritance of coronary artery disease in men: An analysis of the role. Lancet 2012;379(9819):915-22.##Krieglstein CF, Granger DN. Adhesion molecules and their role in vascular disease. Am J Hypertens 2001;14(6Pt 2):44S-54S.##Chan KH, Huang YT, Meng Q, Wu C, Reiner A, Sobel EM, et al. Shared molecular pathways, and gene networks for cardiovascular disease, and type 2 diabetes mellitus in women across diverse ethnicities. Circ Cardiovasc Genet 2014;7(6):911-9. ##Carter AM. Complement activation: an emerging player in the pathogenesis of cardiovascular disease. Scientifica (Cairo) 2012;2012:402783. ##

Role of Phosphorylation and Hyperphosphorylation of Tau in Its Interaction with βα Dimeric Tubulin Studied from a Bioinformatics Perspective 2 2 Background: Tau is a disordered Microtubule Associated Protein (MAP) which prefers to bind and stabilize microtubules. Phosphorylation of tau in particular enhances tau-tubulin interaction which otherwise detaches from tubulin during hyperphosphorylation. The reason behind their destabilization, detachment and the role of β subunit (from microtubule) and the projection domain (Tau) in microtubule stability remains elusive till date. Thus, a complete 3D structural investigation of tau protein is much needed to address these queries as the existing crystal structures are in fragments and quite limited. Methods: In this study, the modelled human tau protein was subjected to phosphorylation and hyperphosphorylation which were later considered for docking with microtubules (βα subunits-inter dimer) and vinblastine. Results: Phosphorylated tau protein interacts with both α- and β subunits. But stronger bonding was with α- compared to β subunits. Regarding β subunit, proline rich loop and projection domain actively participated in tau binding. Interestingly, hyperphosphorylation of tau increases MAP domain flexibility which ultimately results in tau detachment, the main reason behind tangle formation in Alzheimer’s disease. Conclusion: This study being the first of its kind emphasizes the role of projection domain and proline rich region of β-subunit in stabilizing the tau-tubulin interaction and also the effect of hyperphosphorylation in protein-protein and protein-drug binding. 24 34 Hrushikesh Dixit Faculty of Biotechnology and Bioinformatics, D.Y.Patil Deemed to be University, CBD Belapur Faculty of Biotechnology and Bioinformatics, D.Y.Patil Deemed to be University, CBD Belapur India Selvaa Kumar C Ruchi Chaudhary Faculty of Biotechnology and Bioinformatics, D.Y.Patil Deemed to be University, CBD Belapur Faculty of Biotechnology and Bioinformatics, D.Y.Patil Deemed to be University, CBD Belapur India Divya Thaker Faculty of Biotechnology and Bioinformatics, D.Y.Patil Deemed to be University, CBD Belapur Faculty of Biotechnology and Bioinformatics, D.Y.Patil Deemed to be University, CBD Belapur India Nikhil Gadewal Debjani Dasgupta PhosphorylationTau proteinsTubulinVinblastine 40444.pdf Drubin D. Tau protein function in living cells. J Cell Biol 1986;103(6):2739-46. ##Weingarten MD, Lockwood AH, Hwo SY, Kirschner MW. A protein factor essential for microtubule assembly. Proc Natl Acad Sci USA 1975;72(5):1858-62. ##Dehmelt L, Halpain S. The MAP2/Tau family of microtubule-associated proteins. Genome Biol 2004;6(1):204.##Fichou Y, Heyden M, Zaccai G, Weik M, Tobias D. Molecular dynamics simulations of a powder model of the intrinsically disordered protein Tau. J Phys Chem B 2015;119(39):12580-9. ##Goedert M, Jakes R, Spillantini MG, Hasegawa M, Smith MJ, Crowther RA. Assembly of microtubule-associated protein tau into Alzheimer-like filaments induced by sulphated glycosaminoglycans. Nature 1996;383(6600):550-3. ##Pedersen JT, Sigurdsson EM. Tau immunotherapy for Alzheimer's disease. Trends Mol Med 2015;21(6):394-402.##Goode BL, Denis PE, Panda D, Radeke MJ, Miller HP, Wilson L, Feinstein SC. Functional interactions between the proline-rich and repeat regions of tau enhance microtubule binding and assembly. Mol Biol Cell 1997;8(2): 353-65.##Jung D, Filliol D, Miehe M, Rendon A. Interaction of brain mitochondria with microtubules reconstituted from brain tubulin and MAP2 or TAU. Cell Motil Cytoskeleton 1993;24(4):245-55.##Butner K. Tau protein binds to microtubules through a flexible array of distributed weak sites. J Cell Biol 1991;115(3):717-30.##Derisbourg M, Leghay C, Chiappetta G, Fernandez-Gomez FJ, Laurent C, Demeyer D, et al. Role of the Tau N-terminal region in microtubule stabilization revealed by new endogenous truncated forms. Sci Rep 2015;5(1):9659.##Gustke N, Trinczek B, Biernat J, Mandelkow E, Mandelkow E. Domains of tau Protein and interactions with microtubules. Biochemistry 1994;33(32):9511-22.##Mukrasch M, von Bergen M, Biernat J, Fischer D, Griesinger C, Mandelkow E, et al. The “Jaws” of the Tau-Microtubule Interaction. J Biol Chem 2007;282(16):12230-9. ##Ghetti B, Oblak A, Boeve B, Johnson K, Dickerson B, Goedert M. Invited review: Frontotemporal dementia caused by microtubule-associated protein taugene (MAPT) mutations: a chameleon for neuropathology and neuroimaging. Neuropathol Appl Neurobiol 2015;41(1):24-46.##Li J, Shariff A, Wiking M, Lundberg E, Rohde G, Murphy R. Estimating microtubule distributions from 2D immunofluorescence microscopy images reveals differences among human cultured cell lines. PloS One 2012;7(11):e50292.##Hyams JS, Lloyd CW. Microtubules. New York: Wiley-Liss; 1994.##Williams R, Shah C, Sackett D. Separation of tubulin isoforms by isoelectric focusing in immobilized pH gradient gels. Anal Biochem 1999;275(2):265-7. ##Caplow M, Ruhlen RL, Shanks J. The free energy for hydrolysis of a microtubule-bound nucleotide triphosphate is near zero: all of the free energy for hydrolysis is stored in the microtubule lattice [published erratum appears in J Cell Biol 1995 Apr;129(2):549]. J Cell Biol 1994;127(3):779-88.##Löwe J, Li H, Downing K, Nogales E. Refined structure of αβ-tubulin at 3.5 Å resolution. J Mol Biol 2001;313(5):1045-57. ##Mickey B. Rigidity of microtubules is increased by stabilizing agents. J Cell Biol 1995;130(4):909-17.##Correia JJ, Baty LT, Williams Jr RC. Mg2+ dependence of guanine nucleotide binding to tubulin. J Biol Chem 1987;262(36):17278-84.##Amos LA. What tubulin drugs tell us about microtubule structure and dynamics. Semin Cell Dev Biol 2011;22(9):916-26. ##Gigant B, Wang C, Ravelli RBG, Roussi F, Steinmetz MO, Curmi PA, et al. Structural basis for the regulation of tubulin by vinblastine. Nature 2005;435(7041):519-22.##Nogales E, Wolf SG, Downing KH. Structure of the αβ tubulin dimer by electron crystallography. Nature 1998;391(6663):199-203.##Kar S, Fan J, Smith MJ, Goedert M, Amos LA. Repeat motifs of tau bind to the insides of microtubules in the absence of taxol. EMBO J 2003;22(1):70-7. ##Pevalova M, Filipcik P, Novak M, Avila J, Iqbal K. Post-translational modifications of tau protein. Bratisl Lek Listy 2006;107(9-10):346-53.##Hong M, Zhukareva V, Vogelsberg-Ragaglia V, Wszolek Z, Reed L, Miller BI, et al. Mutation-specific functional impairments in distinct tau isoforms of hereditary FTDP-17. Science 1998;282(5395):1914-7. ##Hutton M. Missense and splice site mutations in tau associated with FTDP-17: Multiple pathogenic mechanisms. Neurology 2001;56(11 Suppl 4):S21-5.##Lee VM, Balin BJ, Otvos Jr L, Trojanowski JQ. A68: a major subunit of paired helical filaments and derivatized forms of normal Tau. Science 1991;251(4994):675-8. ##Castro TG, Munteanu FD, Cavaco-Paulo A. Electrostatics of tau protein by molecular dynamics. Biomolecules 2019;9(3):116.##Bairoch A, Apweiler R. The SWISS-PROT protein sequence data bank and its new supplement TREMBL. Nucleic Acids Res 1997;24(1):21-5.##Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ. Basic local alignment search tool. J Mol Biol 1990;215(3):403-10.##Yang J, Zhang Y. Protein structure and function prediction using I‐TASSER. Curr Protoc Bioinformatics 2015;52(1): 5.8.1-5.8.15.##Margreitter C, Petrov D, Zagrovic B. Vienna-PTM web server: a toolkit for MD simulations of protein post-translational modifications. Nucleic Acids Res 2013; 41(Web Server issue):W422-6. ##Lauckner J, Frey P, Geula C. Comparative distribution of tau phosphorylated at Ser262 in pre-tangles and tangles. Neurobiol Aging 2003;24(6):767-76. ##Despres C, Byrne C, Qi H, Cantrelle FX, Huvent I, Chambraud B, et al. Identification of the Tau phosphorylation pattern that drives its aggregation. Proc Natl Acad Sci USA 2017;114(34):9080-5. ##Neddens J, Temmel M, Flunkert S, Kerschbaumer B, Hoeller C, Loefller T, et al. Phosphorylation of different tau sites during progression of Alzheimer’s disease. Acta Neuropathol Commun 2018;6(1):52. ##Šimić G, Babić Leko M, Wray S, Harrington C, Delalle I, Jovanov-Milošević N, et al. Tau protein hyperphosphorylation and aggregation in Alzheimer’s disease and other tauopathies, and possible neuroprotective strategies. Biomolecules 2016;6(1):6. ##Mondragón-Rodríguez S, Perry G, Luna-Muñoz J, Acevedo-Aquino MC, Williams S. Phosphorylation of tau protein at sites Ser396-404is one of the earliest events in Alzheimer's disease and Down syndrome. Neuropathol Appl Neurobiol 2014;40(2):121-35. ##Sandhu P, Naeem M, Lu C, Kumarathasan P, Gomes J, Basak A. Ser422 phosphorylation blocks human Tau cleavage by caspase-3: Biochemical implications to Alzheimer’s Disease. Bioorg Med Chem Lett 2017;27(3):642-52.##Alonso A, Di Clerico J, Li B, Corbo CP, Alaniz ME, Grundke-Iqbal I, et al. Phosphorylation of Tau at Thr212, Thr231, and Ser262 combined causes neurodegeneration. J Biol Chem 2010;285(40):30851-60.##Thakur N. PPredYeast [Internet]. Crdd.osdd.net. 2020 [cited July 19, 2019]. Available from: http://crdd.osdd.net/servers/ppredyeast/help.php ##Berman HM, Westbrook J, Feng Z, Gilliland G, Bhat TN, Weissig H, et al. The protein data bank. Nucleic Acids Res 2000;28(1):235-42. ##Prota AE, Bargsten K, Northcote PT, Marsh M, Altmann KH, Miller JH, et al. Structural basis of microtubule stabilization by laulimalide and peloruside A. Angew Chem (International ed. in English) 2014;53(6):1621-5. ##Guex N, Peitsch MC, Schwede T. Automated comparative protein structure modeling with SWISS-MODEL and Swiss-PdbViewer: A historical perspective. Electrophoresis 2009; 30(S1):S162-73. ##Al-Bassam J, Ozer RS, Safer D, Halpain S, Milligan RA. MAP2 and tau bind longitudinally along the outer ridges of microtubule protofilaments. J Cell Biol 2002;157(7):1187-96.##Welcome to the GROMACS documentation-GROMACS 5.1.5 documentation [Internet]. Manual.gromacs.org. 2020 [cited August ‎8, ‎2019]. Available from: http://manual.gromacs.org/documentation/5.1-current/index.html##Jorgensen WL, Chandrasekhar J, Madura JD. Comparison of simple potential functions for simulating liquid water. J Chem Phys 1983;79(2):926-35.##Berendsen HJC, Postma JPM, van Gunsteren WF, DiNola A, Haak JR. Molecular dynamics with coupling to an external bath. J Chem Phys 1984;81(8):3684-90.##Car R, Parrinello M. Unified approach for molecular dynamics and density-functional theory. Phys Rev Lett 1985;55(22):2471-2474.##Shaw DE. A fast, scalable method for the parallel evaluation of distance-limited pairwise particle interactions. J Comput Chem 2005;26(13):1318-28.##Iu Lobanov M, Bogatyreva NS, Galzitskaya OV. [Radius of gyration as an indicator of protein structure compactness]. Mol Biol (Mosk) 2008;42(4):701-6. Russian. ##Kozakov D, Hall DR, Xia B, Porter KA, Padhorny D, Yueh C, et al. The ClusPro web server for protein–protein docking. Nat Protoc 2017;12(2):255-78. ##Yang Y, Zhou Y. Specific interactions for ab initio folding of protein terminal regions with secondary structures. Proteins 2008;72(2):793-803.##Ranaivoson FM, Gigant B, Berritt S, Joullié M, Knossow M. Structural plasticity of tubulin assembly probed by vinca-domain ligands. Acta Crystallogr D 2012;68(8):927-34. ##Friesner RA, Murphy RB, Repasky MP, Frye LL, Greenwood JR, Thomas A Halgren, et al. Extra precision glide: docking and scoring incorporating a model of hydrophobic enclosure for protein−ligand complexes. J Med Chem 2006;49(21):6177-96.##Fontela YC, Kadavath H, Biernat J, Riedel D, Mandelkow E, Zweckstetter M. Multivalent cross-linking of actin filaments and microtubules through the microtubule-associated protein Tau. Nat Commun 2017;8(1):1981.##Cehlar O, Skrabana R, Novak M. Structure of tau peptide in complex with Tau5 antibody Fab fragment. 2015. ##Pettersen EF, Goddard TD, Huang CC, Couch GS, Greenblatt DM, Meng EC, et al. UCSF Chimera-A visualization system for exploratory research and analysis. J Comput Chem 2004;25(13):1605-12.##Martin L, Latypova X, Terro F. Post-translational modifications of tau protein: Implications for Alzheimer's disease. Neurochem Int 2011;58(4):458-71. ##Goode BL, Denis PE, Panda D, Radeke MJ, Miller HP, Wilson L, et al. Functional interactions between the proline-rich and repeat regions of tau enhance microtubule binding and assembly. Mol Biol Cell 1997;8(2):353-65. ##

Antibiotic Resistance Pattern in Pseudomonas aeruginosa Isolated from Clinical Samples Other than Burn Samples in Iran 2 2 Background: The purpose of this study was to systematically review the prevalence of class 1 integrons, antibiotic resistance pattern in Pseudomonas aeruginosa (P. aeruginosa) isolated from clinical samples other than burn samples. Methods: The Web of Science, PubMed, Scopus, and Science Direct databases were searched using keywords based on the Preferred Reporting Items for Systematic Review and Meta-Analyses (PRISMA) guidelines. The cross-sectional studies published from 1st January 2000 until 1st January 2019 were included which addressed the prevalence of class 1 integrons and antibiotic-resistance in P. aeruginosa isolated from clinical samples other than burn samples. Meta-analysis was conducted using Comprehensive Meta-Analysis (CMA) software. The random-effects model, Cochran’s Q and I2 tests were applied for statistical analyses. Results: Eight articles met the eligibility standards for including in the present meta-analysis. The combined prevalence of class 1 integrons in P. aeruginosa isolated from clinical samples other than burn samples was reported by 40% (95% CI:26.1-55.8%). The pooled prevalence of Multi-Drug Resistant (MDR) P. aeruginosa isolates was 70.1%. The highest prevalence of combined antibiotic resistance was related to carbenicillin with a resistance rate of 79.9%. In general, 6 (75%) out of the 8 included studies showed the correlation between the presence of class 1 integrons and antibiotic resistance. Conclusion: Regarding the correlation between the presence of integrons and the high antibiotic resistance reported by studies included in the present review, there is the need for preventive measures to prevent the spread of resistance by integrons and transferring to other micro-organisms. 35 41 Ebrahim Karimi Emergency Department, Be'sat Hospital, AJA University of Medical Sciences Emergency Department, Be'sat Hospital, AJA University of Medical Sciences Iran Fatemeh Ghalibafan Student Research Committee, Faculty of Medicine, Mashhad University of Medical Sciences Student Research Committee, Faculty of Medicine, Mashhad University of Medical Sciences Iran Akram Esfandani Arak University of Medical Sciences Arak University of Medical Sciences Iran Niusha Manoochehri Arash Faculty of Medicine, Shahid Beheshti University of Medical Sciences Faculty of Medicine, Shahid Beheshti University of Medical Sciences Iran Sassan Mohammadi Faculty of Medicine, Shahid Beheshti University of Medical Sciences Faculty of Medicine, Shahid Beheshti University of Medical Sciences Iran Azad Khaledi Infectious Diseases Research Center, Kashan University of Medical Sciences Infectious Diseases Research Center, Kashan University of Medical Sciences Iran Hakimeh Akbari Maria Khurshid BurnsDrug resistanceIntegronsPseudomonas aeruginosa 40445.pdf Sadikot RT, Blackwell TS, Christman JW, Prince AS. Pathogen–host interactions in Pseudomonas aeruginosa pneumonia. Am J Respir Crit Care Med 2005;171(11):1209-23. ##Mobaraki S, Aghazadeh M, Barhaghi MHS, Memar MY, Goli HR, Gholizadeh P, et al. Prevalence of integrons 1, 2, 3 associated with antibiotic resistance in Pseudomonas aeruginosa isolates from Northwest of Iran. BioMedicine 2018;8(1):2.##Haidar G, Philips NJ, Shields RK, Snyder D, Cheng S, Potoski BA, et al. Ceftolozane-tazobactam for the treatment of multidrug-resistant Pseudomonas aeruginosa infections: clinical effectiveness and evolution of resistance. Clin Infect Dis 2017;65(1):110-20. ##Ruiz-Garbajosa P, Canton R. Epidemiology of antibiotic resistance in Pseudomonas aeruginosa. Implications for empiric and definitive therapy. Rev Esp Quimioter2017;30(Suppl 1):8-12.##Magill SS, Edwards JR, Bamberg W, Beldavs ZG, Dumyati G, Kainer MA, et al. Multistate point-prevalence survey of health care–associated infections. New Engl J Med 2014;370(13):1198-208.##Palomar M, Álvarez-Lerma F, Olaechea P, Insausti J, López-Pueyo M. Estudio nacional de vigilancia de infección nosocomial en servicios de medicina intensiva. SEMICYUC.[Consultado el 10 dic 2014] Disponible en: URL: http://hws. vhebron. net/envin-helics/Help/Informe% 20ENVIN-UCI 202012. ##Vaez H, Salehi-Abargouei A, Ghalehnoo ZR, Khademi F. Multidrug resistant Pseudomonas aeruginosa in Iran: A systematic review and metaanalysis. J Glob Infect Dis 2018;10(4):212-17.##Carattoli A. Importance of integrons in the diffusion of resistance. Veterinary Res 2001;32(3-4):243-59. ##Ruiz-Martínez L, López-Jiménez L, Fusté E, Vinuesa T, Martínez J, Viñas M. Class 1 integrons in environmental and clinical isolates of Pseudomonas aeruginosa. Int J Antimicrob Agents 2011;38(5):398-402.##Gu B, Tong M, Zhao W, Liu G, Ning M, Pan S, et al. Prevalence and characterization of class I integrons among Pseudomonas aeruginosa and Acinetobacter baumannii isolates from patients in Nanjing, China. J Clin Microbiol 2007;45(1):241-3.##Mazel D, Dychinco B, Webb VA, Davies J. Antibiotic resistance in the ECOR collection: integrons and identification of a novel aad gene. Antimicrob Agents Chemother 2000;44(6):1568-74. ##Gillings MR, Gaze WH, Pruden A, Smalla K, Tiedje JM, Zhu YG. Using the class 1 integron-integrase gene as a proxy for anthropogenic pollution. ISME J 2015;9(6):1269-79. ##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(1):45. ##Hall RM, Collis CM. Antibiotic resistance in gram-negative bacteria: the role of gene cassettes and integrons. Drug Resist Updat 1998;1(2):109-19.##Vandenbroucke JP, Von Elm E, Altman DG, Gøtzsche PC, Mulrow CD, Pocock SJ,et al. Strengthening the reporting of Observational Studies in Epidemiology (STROBE): explanation and elaboration. Int J Surg 2014;12(12):1500-24. ##Heidarzadeh S, Enayati Kaliji Y, Pourpaknia R, Mohammadzadeh A, Ghazali-Bina M, Saburi E, et al. A meta-analysis of the prevalence of class 1 integron and correlation with antibiotic resistance in Pseudomonas aeruginosa recovered from Iranian burn patients. J Burn Care Res 2019;40(6):972-8.##Nikokar I, Tishayar A, Flakiyan Z, Alijani K, Rehana-Banisaeed S, Hossinpour M, et al. Antibiotic resistance and frequency of class 1 integrons among Pseudomonas aeruginosa, isolated from burn patients in Guilan, Iran. Iran J Microbiol 2013;5(1):36-41. ##Faghri J, Nouri S, Jalalifar S, Zalipoor M, Halaji M. Investigation of antimicrobial susceptibility, class I and II integrons among Pseudomonas aeruginosa isolates from hospitalized patients in Isfahan, Iran. BMC Res Notes 2018;11(1):806.##Ren CL, Konstan MW, Yegin A, Rasouliyan L, Trzaskoma B, Morgan WJ, et al. Multiple antibiotic-resistant Pseudomonas aeruginosa and lung function decline in patients with cystic fibrosis. J Cystic Fibros 2012;11(4):293-9. ##Cholley P, Thouverez M, Hocquet D, Van Der Mee-Marquet N, Talon D, Bertrand X. Most multidrug-resistant Pseudomonas aeruginosa isolates from hospitals in eastern France belong to a few clonal types. J Clin Microbiol 2011;49(7):2578-83. ##Taccone FS, Cotton F, Roisin S, Vincent JL, Jacobs F. Optimal meropenem concentrations to treat multidrug-resistant Pseudomonas aeruginosa septic shock. Antimicrob Agents Chemother 2012;56(4):2129-31.##Shibata N, Doi Y, Yamane K, Yagi T, Kurokawa H, Shibayama K, et al. PCR typing of genetic determinants for metallo-β-lactamases and integrases carried by gram-negative bacteria isolated in Japan, with focus on the class 3 integron. J Clin Microbiol 2003;41(12):5407-13.##Moradian Kouchaksaraei F, Shahandashti EF, Molana Z, Moradian Kouchaksaraei M, Asgharpour F, Mojtahedi A, et al. Molecular detection of Integron genes and pattern of antibiotic resistance in Pseudomonas aeruginosa strains isolated from intensive care unit, Shahid Beheshti Hospital, North of Iran. Int J Mol Cell Med 2012;1(4):209-17.##Aryanezhad M, Shakibaie MR, Karmostaji A, Shakibaie S. Prevalence of Class 1, 2, and 3 Integrons and Biofilm Formation in Pseudomonas aeruginosa and Acinetobacter baumannii among ICU and non-ICU Patients. Infect Epidemiol Med 2016;2(4):1-7. ##Jabbari Amiri MR, Siami R, Khaledi A. Tuberculosis status and coinfection of pulmonary fungal infections in patients referred to reference laboratory of health centers Ghaemshahr city during 2007-2017. Ethiop J Health Sci 2018;28(6):683-90.##Goli HR, Nahaei MR, Rezaee MA, Hasani A, Kafil HS, Aghazadeh M, et al. Role of MexAB-OprM and MexXY-OprM efflux pumps and class 1 integrons in resistance to antibiotics in burn and Intensive Care Unit isolates of Pseudomonas aeruginosa. J Infect Public Health 2018;11(3):364-72.##Oliver A, Mulet X, Lopez-Causape C, Juan C. The increasing threat of Pseudomonas aeruginosa high-risk clones. Drug Resist Updat 2015;21-22:41-59. ##Chen J, Su Z, Liu Y, Wang S, Dai X, Li Y, et al. Identification and characterization of class 1 integrons among Pseudomonas aeruginosa isolates from patients in Zhenjiang, China. Int J Infect Dis 2009;13(6):717-21. ##Fonseca ÉL, Vieira VV, Cipriano R, Vicente AC. Class 1 integrons in Pseudomonas aeruginosa isolates from clinical settings in Amazon region, Brazil. FEMS Immunol Med Microbiol 2005;44(3):303-9.##Fernández L, Álvarez-Ortega C, Wiegand I, Olivares J, Kocíncová D, Lam JS, et al. Characterization of the polymyxin B resistome of Pseudomonas aeruginosa. Antimicrob Agents Chemother 2013;57(1):110-9.##Gelband H, Miller-Petrie M, Pant S, Gandra S, Levinson J, Barter D, et al. The state of the world's antibiotics 2015. Center for Disease Dynamics, Economics & Policy 2015. CDDEP: Washington, D.C. ##Gales A, Jones R, Turnidge J, Rennie R, Ramphal R. Characterization of Pseudomonas aeruginosa isolates: occurrence rates, antimicrobial susceptibility patterns, and molecular typing in the global SENTRY Antimicrobial Surveillance Program, 1997–1999. Clin Infect Dis 2001;32(Supplement_2):S146-S155.##Bouza E, Garcia-Garrote F, Cercenado E, Marin M, Diaz M. Pseudomonas aeruginosa: a survey of resistance in 136 hospitals in Spain. Antimicrob Agents Chemother 1999;43(4):981-2.##Brown PD, Izundu A. Antibiotic resistance in clinical isolates of Pseudomonas aeruginosa in Jamaica. Rev Panam Salud Publica 2004;16(2):125-30.##Poonsuk K, Tribuddharat C, Chuanchuen R. 2012. Class 1 integrons in Pseudomonas aeruginosa and Acinetobacter baumannii isolated from clinical isolates. Southeast Asian J Trop Med Public Health 2012;43(2):376-84.##Mohammadzadeh A, Mardaneh J, Ahmadi R, Adabi J. Evaluation of the virulence features and antibiotic resistance patterns of pathogenic Pseudomonas aeruginosa strains isolated from hospitalized patients in Gonabad, Iran. Arch Pediatr Infect Dis 2017;5(3):e41267.##Pournajaf A, Razavi S, Irajian G, Ardebili A, Erfani Y, Solgi S, et al. Kafshgari, R. 2018. Integron types, antimicrobial resistance genes, virulence gene profile, alginate production and biofilm formation in Iranian cystic fibrosis Pseudomonas aeruginosa isolates. Infez Med 2018;26(3):226-36.##Salimizadeh Z, Karouei H, Masoud S, Hosseini F. Dissemination of Class 1 integron among different multidrug resistant Pseudomonas aeruginosa strains. Medical Laboratory Journal 2018;12(4):36-42.##

β-sitosterol Mediated Silver Nanoparticles Induce Cytotoxicity in Human Colon Cancer HT-29 Cells 2 2 Background: Silver nanoparticles (AgNP) are commonly used metallic nanoparticles in health care systems. Colon cancer incidence is increasing worldwide. In this study, AgNP was synthesized using β-sitosterol and its cytotoxic potential was evaluated in human colon cancer (HT-29) cells. Methods: Characterization of AgNP was analyzed by TEM and spectrophotometry analysis. HT-29 cells were treated with different concentrations (2, 4, 6, 8 and 10 ng/ml) of AgNPs and cytotoxicity was evaluated by MTT assay. The apoptosis was analyzed by the flow cytometry. The expression of p53 protein was analyzed by western blotting. Results: β-sitosterol mediated AgNP are spherical in shape and induced concentration-dependent cytotoxicity in HT-29 cells. AgNP caused apoptosis related morphological changes as evidenced by annexin positive staining. AgNP treatments also induced the p53 expression in HT-29 cells. Conclusion: Our present result suggests that β-sitosterol mediated AgNP induce apoptosis in colon cancer cells and this finding may pave the way for further experimental analysis in vivo. 42 46 Palaniappan Chithambara Shathviha Department of Pharmacology, Saveetha Dental College, Saveetha Institute of Medical and Technical Sciences (SIMATS), Chennai Department of Pharmacology, Saveetha Dental College, Saveetha Institute of Medical and Technical Sciences (SIMATS), Chennai India Devaraj Ezhilarasan Shanmugam Rajeshkumar Department of Pharmacology, Saveetha Dental College, Saveetha Institute of Medical and Technical Sciences (SIMATS), Chennai, Department of Pharmacology, Saveetha Dental College, Saveetha Institute of Medical and Technical Sciences (SIMATS), Chennai, India Jayaraman Selvaraj Department of Biochemistry, Saveetha Dental College (SDC), Saveetha Institute of Medical and Technical Sciences, Chennai Department of Biochemistry, Saveetha Dental College (SDC), Saveetha Institute of Medical and Technical Sciences, Chennai India ApoptosisColon neoplasms HT29 cellsSilver nanoparticlesβ-sitosterol 40446.pdf Jahangirian H, Kalantari K, Izadiyan Z, Rafiee-Moghaddam R, Shameli K, Webster TJ. A review of small molecules and drug delivery applications using gold and iron nanoparticles. Int J Nanomedicine 2019;14:1633-57.##Shi J, Kantoff PW, Wooster R, Farokhzad OC. Cancer nanomedicine: progress, challenges and opportunities. Nat Rev Cancer 2017;17(1):20-37. ##Vijaya Yadav AK. In vitro anthelmintic assessment of selected phytochemicals against Hymenolepis diminuta, a zoonotic tapeworm. J Parasit Dis 2016;40(3):1082-6.##Shi C, Wu F, Zhu XC, Xu J. Incorporation of beta-sitosterol into the membrane increases resistance to oxidative stress and lipid peroxidation via estrogen receptor-mediated PI3K/GSK3beta signaling. Biochim Biophys Acta 2013;1830(3):2538-44.##PonselviInduja M, Ezhilarasan D, Ashok Vardhan N. Evolvulus alsinoides methanolic extract triggers apoptosis in HepG2 cells. Avicenna J Phytomed 2018;8(6):504-12.##Gurunathan S, Kang MH, Kim JH. Combination effect of silver nanoparticles and histone deacetylases inhibitor in human alveolar basal epithelial cells. Molecules 2018;23(8):E2046.##Kummrow A, Frankowski M, Bock N, Werner C, Dziekan T, Neukammer J. Quantitative assessment of cell viability based on flow cytometry and microscopy. Cytometry A 2013;83(2):197-204.##Akther T, Mathipi V, Kumar NS, Davoodbasha MA, Srinivasan H. Fungal-mediated synthesis of pharmaceutically active silver nanoparticles and anticancer property against A549 cells through apoptosis. Environ Sci Pollut Res Int 2019;26(13):13649-57.##Rohit Singh T, Ezhilarasan D. Ethanolic extract of Lagerstroemia speciosa (L.) Pers., induces apoptosis and cell cycle arrest in HepG2 cells. Nutr Cancer 2020;72(1):146-56.##Kovács D, Igaz N, Keskeny C, Bélteky P, Tóth T, Gáspár R, et al. Silver nanoparticles defeat p53-positive and p53-negative osteosarcoma cells by triggering mitochondrial stress and apoptosis. Sci Rep 2016;6:27902.##

Comparison of Antifungal Properties of Gold, Silver, and Selenium Nanoparticles Against Amphotericin B-Resistant Candida glabrata Clinical Isolates 2 2 Background: The present study aimed to investigate the antifungal activity of Nanoparticles (NPs) against amphotericin B-resistant Candida glabrata (C. glabrata) strains. Methods: Twelve resistant (C. glabrata) strains were isolated from archived clinical isolates. Antifungal activity was conducted according to Clinical and Laboratory Standards Institute’s (CLSI) guidelines, document M27-A3/S4. The Scanning Electron Microscope (SEM) was used to observe the morphological changes of strains exposed to each nanoparticle. Results: Minimum Inhibitory Concentration (MIC) of nanoparticles of all strains was in the concentration range of 0.125 to 0.5 µg/Ml. The synthesized Ag-NPs showed superior antifungal activity against (C. glabrata) compared to Se-NPs and Au-NPs. The scanning electron microscope images revealed the difference in the fungal morphology between the untreated and treated fungi with nanoparticles. Conclusion: The Ag-NPs, followed by Se-NPs synthesized, revealed significant antifungal activity against resistance regardless of their antifungal-resistant mechanisms. 47 50 Ensieh Lotfali Department of Medical Parasitology and Mycology, Faculty of Medicine, Shahid Beheshti University of Medical Sciences Department of Medical Parasitology and Mycology, Faculty of Medicine, Shahid Beheshti University of Medical Sciences Iran Hossein Toreyhi Student Research Committee, Faculty of Medicine, Shahid Beheshti University of Medical Sciences Student Research Committee, Faculty of Medicine, Shahid Beheshti University of Medical Sciences Iran Kamyab Makhdoomi Sharabiani Student Research Committee, Faculty of Medicine, Shahid Beheshti University of Medical Sciences Student Research Committee, Faculty of Medicine, Shahid Beheshti University of Medical Sciences Iran Azam Fattahi Amirali Soheili Student Research Committee, Faculty of Medicine, Shahid Beheshti University of Medical Sciences Student Research Committee, Faculty of Medicine, Shahid Beheshti University of Medical Sciences Iran Reza Ghasemi Student Research Committee, Faculty of Medicine, Shahid Beheshti University of Medical Sciences Student Research Committee, Faculty of Medicine, Shahid Beheshti University of Medical Sciences Iran Mahyar Keymaram Department of Medical Parasitology and Mycology, Faculty of Medicine, Iran University of Medical Sciences Department of Medical Parasitology and Mycology, Faculty of Medicine, Iran University of Medical Sciences Iran Yasaman Rezaee Student Research Committee, Faculty of Pharmacy, Shahid Beheshti University of Medical Sciences Student Research Committee, Faculty of Pharmacy, Shahid Beheshti University of Medical Sciences Iran Sayna Iranpanah Student Research Committee, Faculty of Pharmacy, Shahid Beheshti University of Medical Sciences Student Research Committee, Faculty of Pharmacy, Shahid Beheshti University of Medical Sciences Iran Antifungal agentsCandida glabrataNanoparticlesSilver 40447.pdf Geffers C, Gastmeier P. Nosocomial infections and multidrug-resistant organisms in Germany: epidemiological data from KISS (The Hospital Infection Surveillance System). Dtsch Arztebl Int 2011;108(6):87-93.##Silva S, Negri M, Henriques M, Oliveira R, Williams DW, Azeredo J. Candida glabrata, Candida parapsilosis and Candida tropicalis: biology, epidemiology, pathogenicity and antifungal resistance. FEMS Microbiol Rev 2012;36(2):288-305.##Dos Santos CA, Seckler MM, Ingle AP, Gupta I, Galdiero S, Galdiero M, et al. Silver nanoparticles: therapeutical uses, toxicity, and safety issues. J Pharm Sci 2014;103(7):1931-44.##Clinical, Institute LS. Reference method for broth dilution antifungal susceptibility testing of yeasts. Clinical and Laboratory Standards Institute Wayne, PA; 2008.##Alimehr Sh, Abad HSE, Shahverdi AR, Hashemi J, Zomorodian K, Moazeni M, et al. Comparison of difference between fluconazole and silver nanoparticles in antimicrobial effect on fluconazole-resistant Candida Albicans strains. Arch Pediatr Infect Dis 2015;3(2):e21481.##Nasrollahi A, Pourshamsian K, Mansourkiaee P. Antifungal activity of silver nanoparticles on some of fungi. Int J Nano Dimension 2011;1(3):233-9.##Kim KJ, Sung WS, Moon SK, Choi JS, Kim JG, Lee DG. Antifungal effect of silver nanoparticles on dermatophytes. J Microbiol Biotechnol 2008;18(8):1482-4. ##Sharma VK, Yngard RA, Lin Y. Silver nanoparticles: green synthesis and their antimicrobial activities. Adv Colloid Interface Sci 2009;145(1-2):83-96.##Vijayakumar M, Noorlidah A, Ahmed ABA, Nancy FT, Priya K. Biosynthesis, characterisation and anti-bacterial effect of plant-mediated silver nanoparticles using Artemisia nilagirica. Industrial Crops and Products 2013;41:235-40.##Rónavári A, Igaz N, Gopisetty MK, Szerencsés B, Kovács D, Papp C, et al. Biosynthesized silver and gold nanoparticles are potent antimycotics against opportunistic pathogenic yeasts and dermatophytes. Int J Nanomedicine 2018;13:695.##

An Evaluation of Transmission Dynamics of Cryptosporidium Using Molecular Methods 2 2 Dear Editor-in-Chief, The members of genus Cryptosporidium are intracellular parasites that infect mammals, poultry, reptiles and amphibians. From the total of 30 valid species mentioned currently, 14 have been determined to infect the human being. Two species, Cryptosporidium hominis (C. hominis) (Anthroponotic species) and Cryptosporidium parvum (C. parvum) bovine genotype (Zoonotic species), are considered of major public health importance 1. Commonly, the ubiquitous oocysts of Cryptosporidium are transmitted via direct contact with infected hosts or indirectly via contaminated food and water 2. The low infectious dose and its resistance against common water disinfectants make it a challenge for the drinking water plants 3. Common laboratory techniques which are being used for diagnosis of Cryptosporidium cannot discriminate it at species and genotype level. However, the genetic tools allow species determination of the parasite as well as tracing its transmission routes 2. In this study, a total of 55 drinking water samples were collected from 11 different areas of Tabriz, the largest city in North West of Iran. Each sample contained 30 L of water. To collect the suspended particles, samples were filtered through a membrane filter with 1.2 μm pore size (Sartorius, Germany). The pellets trapped on the filter were collected 4. All water pellets were subjected to DNA extraction by a method described previously 5. Then, the amplification of small ribosomal subunit RNA (SSU-rRNA; 18S rRNA) gene was done using a two step nested PCR method 6. A sample of C. parvum DNA that was extracted by the extraction method was included in each round of PCR as a positive control. The multi copy nature of 18S rRNA gene and nested format of the PCR make it a very sensitive method for detection of Cryptosporidium oocysts in water samples 7. However, the expected amplicon (826-864-bp) was not detected in any of the water samples. Various reports from many areas of the world provide strong evidence that contaminated water is an important risk factor for cryptosporidiosis 8. Several factors, including water source, location of sampling, number and volumes of samples, type of ecosystem, climate and detecting procedures are effective for detection of Cryptosporidium oocysts in the water samples 9. The drinking water supplied from surface water sources is more susceptible to contamination. Thus, underground water is a more protected source 10. In our study area, the main part of the drinking water supplies comes from rivers. The role of rainfall as a determining risk factor for the waterborne transmission of Cryptosporidium can be significant. The relationship between increased rainfall and an increase in the concentration of Cryptosporidium oocysts in nearby river waters has been reported 10. The North West part of Iran is an area with lower than average rainfall, which could be the most important reason for the lack of parasites in the water. There are not many reports about the prevalence of Cryptosporidium in water samples in Iran. Meanwhile, the prevalence of Cryptosporidium on the surface and recreational water was 36 and 20%, respectively 4,9. The two researches conducted in Ardabil and Chaharmahal va Bakhtiyari provinces as well as our study are limited studies with small sample size. Thus, more comprehensive studies with large samples from different sources and in different seasons are required to assess the real risk of waterborne cryptosporidiosis in Iran.  Previous studies in Tabriz showed that 1.76% of diarrheic children and 3.8% of cattle have been infected with C. parvum 2,6 (Table 1). The presence of C. parvumm in children, as a sensitive group and in cattle, as a major source for zoonotic disease may be associated with zoonotic transmission of the parasite in the study area. Lack of parasite in drinking water may indicate that this cannot be an important route for transmission; instead, it can be a reason for the low prevalence of the infection in children. Lack of C. hominis (Anthroponotic species) in children and the prevalence of C. parvum, potentially zoonotic species, in cattle and its presence in diarrheic rural children would raise the possibility that zoonotic transmission originally occurs through direct contact with farm animals in this region. Therefore, cattle and other domestic animals should be considered as important sources of infection in the North-western part of Iran. 51 52 Behroz Mahdavi Poor Department of Laboratory Science, Faculty of Paramedicine, Tabriz University of Medical Sciences Department of Laboratory Science, Faculty of Paramedicine, Tabriz University of Medical Sciences Iran Jalil Rashedi Mohammad Asgharzadeh Biotechnology Research Center Faculty of Paramedicine, Tabriz University of Medical Sciences Biotechnology Research Center Faculty of Paramedicine, Tabriz University of Medical Sciences Iran Esmaeel Fallah Faculty of Medicine, Islamic Azad University, Ardabil Branch Faculty of Medicine, Islamic Azad University, Ardabil Branch Iran Editorial 40448.pdf Šlapeta J. Cryptosporidiosis and Cryptosporidium species in animals and humans: a thirty colour rainbow? Int J Parasitol 2013;43(12-13):957-70.##Poor BM, Rashedi J, Asgharzadeh M, Fallah E, Hatam-Nahavandi K, Dalimi A . Molecular characterization of Cryptosporidium species in children with diarrhea in north west of Iran. Int J Mol Cell Med 2015;4(4):235-9.##Fuchslin H, Kotzch S, Egli T. Cryptosporidium spp. in drinking water samples from rural sites in Switzerland. Swiss Med Wkly 2012;142:w13683.##Ghalebin BM, Fallah E, Asgharzadh M, Kazemi AH, Arzanlou M. Detection and identification of Cryptosporidium species in water samples from a river in Ardabil city, northwestern Iran. Res J Biological Sciences 2007;2:498-502.##Nahavandi KH, Fallah E, Asgharzadeh M, Mirsamadi N, Mahdavipour B. Glutamate dehydrogenase and triose-phosphate-isomerase coding genes for detection and genetic characterization of Giardia lamblia in human feces by PCR and PCR-RFLP. Turk J Med Sci 2011;41(2):283-9.##Fallah E, Mahdavi Poor B, Jamali R, Hatam-Nahavandi K, Asgharzadeh M. Molecular characterization of Cryptosporidium isolates from cattle in a slaughterhouse in Tabriz, Northwestern Iran. J Biol Sci 2008;8(3):639-43.##Xiao L. Molecular epidemiology of cryptosporidiosis: An update. Exp Parasitol 2010;124(1):80-9.##Fayer R, Morgan U, Upton SJ. Epidemiology of Cryptosporidium: transmission, detection and identification. Int J Parasitol 2000;30(12-13):1305-22. ##Naeini KM, Asadi M, Chaleshtori MH. Detection and molecular characterization of Cryptosporidium species in recreational waters of Chaharmahal va Bakhtiyari province of Iran using nested-PCR-RFLP. Iran J Parasitol 2011;6(1):20-7. ##Rose JB, Huffman DE, Gennaccaro A. Risk and control of waterborne cryptosporidiosis. FEMS Microbiol Rev 2002;26(2):113-23.##