Shiraz University PressMolecular Biology Research Communications2322-181X10320210901Prokaryotic expression of chimeric GFP-hFc protein as a potential immune-based tool105108615410.22099/mbrc.2021.39728.1588ENThanh-Tan NguyenDepartment of Molecular and Environmental Biotechnology, Faculty of Biology and Biotechnology, University of Science, Ho Chi Minh City, VietnamVietnam National University, Ho Chi Minh City, Vietnam0000-0001-6305-0468Hai-Vy Vo-NguyenDepartment of Molecular and Environmental Biotechnology, Faculty of Biology and Biotechnology, University of Science, Ho Chi Minh City, VietnamVietnam National University, Ho Chi Minh City, Vietnam0000-0001-8422-6098Hieu Tran-VanDepartment of Molecular and Environmental Biotechnology, Faculty of Biology and Biotechnology, University of Science, Ho Chi Minh City, VietnamVietnam National University, Ho Chi Minh City, Vietnam0000-0003-2782-5232Journal Article20210208GFP is an old-yet-powerful protein marker, which has been widely used in molecular biotechnology due to its capacity of exhibiting bright green fluorescence when exposed to ultraviolet light. The hFc region of IgG antibodies is a specific binding ligand of expressed receptors on immune cells with well-known cellular-associated functions like opsonization and phagocytosis. In this present study, we proceeded to fuse <em>gfp-hfc</em> gene into pET-28a to create a recombinant pET-28a-gfp-hfc vector. The expression of GPF-hFc was induced by IPTG and confirmed using SDS-PAGE and followed by Western blot probed with 6xHis antibodies. This chimeric protein was utilized in specific binding experiments with protein A/G-coated magnetic beads using a fluorescence microscope. Due to its fluorescence and binding ability, GFP-hFc could be used as a model molecule for monitoring molecule detection studies, tracking nanoparticle migration and distribution, or stimulating immune responses.https://mbrc.shirazu.ac.ir/article_6154_ad0b8cd1539f3ec975fd621061d41684.pdfShiraz University PressMolecular Biology Research Communications2322-181X10320210901Role of SALL4 and Nodal in the prognosis and tamoxifen resistance of estrogen receptor-positive breast cancer109119618410.22099/mbrc.2021.39878.1597ENArad BoustanDepartment of Medical Biotechnology and Nanotechnology, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, IranFatemeh MosaffaBiotechnology Research Center, Pharmaceutical Technology Institute, Mashhad University of Medical Sciences, Mashhad, IranDepartment of Pharmaceutical Biotechnology, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, IranRosa JahangiriDepartment of Medical Biotechnology and Nanotechnology, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, IranHamid Heidarian-MiriDepartment of Epidemiology, School of Health, Mashhad University of Medical Sciences, Mashhad, IranAsefeh Dahmardeh-GhalehnoDepartment of Pharmaceutical Biotechnology, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, IranKhadijeh JamialahmadiBiotechnology Research Center, Pharmaceutical Technology Institute, Mashhad University of Medical Sciences, Mashhad, IranJournal Article20210221Despite the discovery of a number of different mechanisms underlying tamoxifen resistance, its molecular pathway is not completely clear. The upregulation of <em>SALL4</em> and <em>Nodal</em> has been reported in breast cancer. Nevertheless, their role in tamoxifen resistance has not been investigated. In the present study, we compared <em>Nodal</em> and <em>SALL4</em> expression in 72 tamoxifen sensitive (TAMS) and tamoxifen-resistant (TAMR) patients. Afterward, the correlation of expression data with clinicopathological features and survival of patients was studied. Results showed that both <em>SALL4</em> and <em>Nodal</em> were significantly upregulated in TAMR compared to TAMS patients. Besides, there was a positive association between <em>Nodal</em> and <em>SALL4</em> expression. Furthermore, we evaluated their correlation with the expression of <em>Oct4</em>, <em>Nanog</em> and <em>Sox2</em> stemness markers. The results demonstrated that in most tissue samples there was a positive correlation between <em>Nodal</em> and <em>SALL4 </em>expression with these stemness markers. Besides, the overexpression of <em>SALL4</em> and <em>Nodal</em> significantly correlated with the N stage. Moreover, the overexpression of <em>SALL4</em> was associated with extracapsular invasion and lymphatic invasion. High level expressions of <em>SALL4</em> and <em>Nodal</em> had a significant association with worse disease-free survival (DFS) rates. In addition, increased level of <em>Nodal</em> expression provides a superior predictor factor for DFS. The multivariate Cox regression analysis also revealed that for DFS, perineural invasion (PNI) was independently an unfavorable prognostic value. These findings suggest that the high expression of <em>SALL4</em> and <em>Nodal</em> could contribute to tamoxifen resistance and worse survival rates in tamoxifen-treated ER<sup>+</sup> breast cancer patients.https://mbrc.shirazu.ac.ir/article_6184_f0c196e60525f0a5d4d4dfd5fe5fa789.pdfShiraz University PressMolecular Biology Research Communications2322-181X10320210901Telomeric zinc-finger associated protein (TZAP) in cancer biology: friend or foe?121129619010.22099/mbrc.2021.40106.1607ENGabriel ArantesDos SantosUrology Department, Laboratory of Medical Investigation (LIM55), Faculdade de Medicina FMUSP, Universidade de Sao Paulo, Sao Paulo, SP, BrazilD'Or Institute for Research and Education (IDOR), Sao Paulo, Brazil0000-0003-1602-9193Nayara IzabelVianaUrology Department, Laboratory of Medical Investigation (LIM55), Faculdade de Medicina FMUSP, Universidade de Sao Paulo, Sao Paulo, SP, BrazilMinas Gerais State University (UEMG), Passos, Minas Gerais, Brazil0000-0003-4505-8508Ruan PimentaUrology Department, Laboratory of Medical Investigation (LIM55), Faculdade de Medicina FMUSP, Universidade de Sao Paulo, Sao Paulo, SP, BrazilD'Or Institute for Research and Education (IDOR), Sao Paulo, Brazil0000-0002-3423-5647Juliana AlvesDe CamargoUrology Department, Laboratory of Medical Investigation (LIM55), Faculdade de Medicina FMUSP, Universidade de Sao Paulo, Sao Paulo, SP, Brazil0000-0001-5624-3798Sabrina T.ReisUrology Department, Laboratory of Medical Investigation (LIM55), Faculdade de Medicina FMUSP, Universidade de Sao Paulo, Sao Paulo, SP, Brazil Athens University Center (UniAtenas), Passos, Minas Gerais, Brazil0000-0002-3564-3597Katia Ramos MoreiraLeiteUrology Department, Laboratory of Medical Investigation (LIM55), Faculdade de Medicina FMUSP, Universidade de Sao Paulo, Sao Paulo, SP, Brazil0000-0002-2615-7730Miguel SrougiUrology Department, Laboratory of Medical Investigation (LIM55), Faculdade de Medicina FMUSP, Universidade de Sao Paulo, Sao Paulo, SP, BrazilD'Or Institute for Research and Education (IDOR), Sao Paulo, Brazil0000-0002-4545-0596Journal Article20210317The new identified protein telomeric zinc-finger associated protein (TZAP) is a negative regulator of telomere length. Since telomere length and telomere maintenance mechanisms are essential to cancer progression, TZAP is considered a new player in cancer biology. Here we aimed to analyze TZAP using the Cancer Genome Atlas data in a Pan-Cancer approach. We gathering data from TCGA Pan-Cancer studies utilizing cBioPortal, GEPIA and UALCAN. In total we analyzed 33 types of cancer (n=9664) and their respective controls (n=711). TZAP is transcribed in all cancers but less than 5% of all tumors show any somatic changes. TZAP was downregulated in kidney chromophobe carcinoma, and upregulated in esophageal cancer, head and neck squamous cell carcinomas, kidney renal clear cell carcinoma and in liver hepatocellular carcinoma. Globally, TZAP expression is related to favorable prognosis, associated to better overall and disease-free survival. Looking to specific tumors, TZAP expression has a dual behavior. Its downregulation is associated with poor prognosis in cervical squamous cell carcinoma, in kidney renal clear cell carcinoma, kidney papillary cell carcinoma, lung adenocarcinoma and pancreas adenocarcinoma. On the contrary, in adrenocortical carcinoma, colon and rectal cancer, brain lower grade glioma and prostate adenocarcinoma the upregulation of TZAP is related with poor prognosis. TZAP expression has a positive correlation with TRF1 and TRF2 in normal tissue but not in cancer. Our analyses indicate that TZAP has an important role in oncology and may be considered as a potential biomarker.https://mbrc.shirazu.ac.ir/article_6190_3eb7ebcaea991e8a1594ad89663ea631.pdfShiraz University PressMolecular Biology Research Communications2322-181X10320210901In silico identification of promising inhibitor against RNA-dependent RNA polymerase target of SARS-CoV-2131140622910.22099/mbrc.2021.40367.1621ENPushpendra SinghState Virus Research and Diagnostic Laboratory, Department of Microbiology, All India Institute of Medical Sciences, Raipur, Chhattisgarh-492099 India0000-0002-9619-0659Manish KumarTripathiDepartment of Biophysics, All India Institute of Medical Sciences, New Delhi-110029, IndiaMohammad YasirDepartment of Nephrology, All India Institute of Medical Science Bhopal, Madhya Pradesh-462020 IndiaRuchi KhareDepartment of Biological Science and Engineering, Maulana Azad National Institute of Technology, Bhopal, Madhya Pradesh-462003 IndiaRahul ShrivastavaDepartment of Biological Science and Engineering, Maulana Azad National Institute of Technology, Bhopal, Madhya Pradesh-462003 IndiaJournal Article20210415The severe acute respiratory syndrome is a viral respiratory disease recognised as COVID-19, caused by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2). Formerly, no precise remedies are available, and many studies regarding COVID-19 prevention and treatment are under development. Several targets for the design of drugs are identified, and studies are in headway to explore the potential target. RNA-dependent RNA polymerase (RdRp) protein identified as a promising target against SARS-CoV-2 infection for the drug design due to its significant role in viral replication. The present study focuses on identifying the binding effect of previously known RdRp inhibitors with RdRp of SARS-CoV-2 using molecular docking and molecular dynamics simulation approaches. Molecular docking and binding free energy calculations against RdRp enzyme identified suramin as a potential compound that showed the highest docking score of -7.83 Kcal/mole and binding energy of -80.83 Kcal/mole as a comparison to other compounds. Further, molecular dynamics simulation studies were moreover showed the stable binding behaviour of suramin docked complex in the protein active site. Thus, the study concludes that suramin might be helpful as a potential inhibitor against RNA-dependent RNA polymerase of SRAS-CoV-2. However, further investigation is needed to assess the possible effect of inhibitors on RdRp through <em>in vitro</em> and <em>in vivo</em> experiments.https://mbrc.shirazu.ac.ir/article_6229_21acb05f56e607c87c53fdf844660486.pdfShiraz University PressMolecular Biology Research Communications2322-181X10320210901MiR-200c-3p expression may be associated with worsening of the clinical course of patients with COVID-19141147623010.22099/mbrc.2021.40555.1631ENRuan PimentaLaboratory of Medical Investigation (LIM55), Urology Department, Faculdade de Medicina da Universidade de São Paulo (FMUSP), São Paulo, Brazil0000-0002-3423-5647Nayara IVianaLaboratory of Medical Investigation (LIM55), Urology Department, Faculdade de Medicina da Universidade de São Paulo (FMUSP), São Paulo, Brazil0000-0003-4505-8508Gabriel ADos SantosLaboratory of Medical Investigation (LIM55), Urology Department, Faculdade de Medicina da Universidade de São Paulo (FMUSP), São Paulo, Brazil0000-0003-1602-9193Patricia CandidoLaboratory of Medical Investigation (LIM55), Urology Department, Faculdade de Medicina da Universidade de São Paulo (FMUSP), São Paulo, Brazil0000-0001-5887-5617Vanessa RGuimaraesLaboratory of Medical Investigation (LIM55), Urology Department, Faculdade de Medicina da Universidade de São Paulo (FMUSP), São Paulo, Brazil0000-0003-3021-722XPoliana RomaoLaboratory of Medical Investigation (LIM55), Urology Department, Faculdade de Medicina da Universidade de São Paulo (FMUSP), São Paulo, Brazil0000-0003-0410-2349Iran ASilvaLaboratory of Medical Investigation (LIM55), Urology Department, Faculdade de Medicina da Universidade de São Paulo (FMUSP), São Paulo, Brazil0000-0001-5441-790XJuliana ADe CamargoLaboratory of Medical Investigation (LIM55), Urology Department, Faculdade de Medicina da Universidade de São Paulo (FMUSP), São Paulo, Brazil0000-0001-5624-3798Dina MHatanakaMoriah Hospital, São Paulo, SP, Brazil0000-0002-3505-5656Paula G SQueirozMoriah Hospital, São Paulo, SP, Brazil0000-0002-0059-9127Alexandre TeruyaMoriah Hospital, São Paulo, SP, Brazil0000-0003-3137-9474Leandro EcheniqueMoriah Hospital, São Paulo, SP, Brazil0000-0001-5182-2192Bruno A M PBesenInstituto Central, Hospital das Clinicas HCFMUSP, Faculdade de Medicina, Universidade de SaoPaulo, Sao Paulo, SP, Brazil0000-0002-3516-9696Katia R MLeiteLaboratory of Medical Investigation (LIM55), Urology Department, Faculdade de Medicina da Universidade de São Paulo (FMUSP), São Paulo, Brazil0000-0002-2615-7730Victor SrougiLaboratory of Medical Investigation (LIM55), Urology Department, Faculdade de Medicina da Universidade de São Paulo (FMUSP), São Paulo, Brazil0000-0001-8346-3833Miguel SrougiLaboratory of Medical Investigation (LIM55), Urology Department, Faculdade de Medicina da Universidade de São Paulo (FMUSP), São Paulo, BrazilMoriah Hospital, São Paulo, SP, Brazil0000-0002-4545-0596Sabrina TReisLaboratory of Medical Investigation (LIM55), Urology Department, Faculdade de Medicina da Universidade de São Paulo (FMUSP), São Paulo, BrazilMoriah Hospital, São Paulo, SP, Brazil0000-0002-3564-3597Journal Article20210503COVID-19 represents a public health emergency, whose mechanism of which is not fully understood. It is speculated that microRNAs may play a crucial role in host cells after infection by SARS-CoV-2. Thus, our study aimed to analyze the expression of miR-200c-3p in saliva samples from patients with COVID-19. One handred eleven samples from patients with COVID-19 were divided into 4 groups. Group I: 39 patients negative for Covid-19; Group II: 37 positive and symptomatic patients, with no indication of hospitalization; Group III: 21 patients with respiratory disorders (hospitalized); Group IV: 14 patients with severe conditions (oxygen therapy). The expression levels of miR-200c-3p were determined using qPCR. We found greater expression of miR-200c-3p in patients in group IV (<em>p </em><0.0001), and also verified that patients aged ≥42 years had a higher expression of this miR (<em>p </em>=0.013). Logistic regression analysis revealed that the expression of miR-200c-3p and systemic arterial hypertension are factors independently associated with patients in group IV (<em>p </em><0.0001). Our results suggest that miR-200c-3p is a predictor of severity independent of COVID-19 risk factors, which could represent a way of screening patients affected by SARS-CoV-2.https://mbrc.shirazu.ac.ir/article_6230_11dc053fbd37be7148fa201ba08a2129.pdf