Overexpression of microRNA-630 in Acute Leukemic T-cell line

Background: MicroRNAs (miRNAs) are noncoding RNAs that control the expression of their target mRNAs. It affects cancer cell proliferation and apoptosis as oncogenes or tumor suppressors. Dysregulation of miRNAs expression leads to the development of various cancers. Therefore, for the first time in this field, this study investigated the effect of overexpression of microRNA-630 on the Jurkat cells. Materials and methods:: In this experimental study, the Jurkat cells were divided into the four groups, i.e. non-transfected control group (A), scramble (B), transfected with 50 nM concentrations of miR-630 (C), and treated with 100 nM miR-630 (D). MiR-630 transfection was performed by lipofectamine 2000. Cancer cell growth in each group was analyzed with MTT assay. Flow cytometry investigated percent of viable, necrotic, and apoptotic cells. Quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) measured the expression of P53, P21, and BCL2 genes. SPSS (version 21) especially Kruskal-Wallis and Mann-Whitney U tests were utilized for data analysis. Results: The results of MTT assay showed that the cell growth rates in C (118%) and D (136%) groups were significantly higher than that in the control group (P= 0.037 vs. 0.034). The percentage of early and late apoptosis in C (3.1% P=0.01, 4.2% P=0.02) and D (0.5% P=0.008, 0.4% P=0.006) groups were significantly lower than those in the control group. The expression of p53 and p21 in C (0.7 P=0.037, 0.62 P=0.034) and D (0.44 P=0.034, 0.53 P=0.038) Groups were significantly decreased compared with the control group. The expression of B-cell lymphoma-2 (Bcl2) in C (1.85) and D (3.26) groups were significantly increased compared with the control group (P= 0.037 vs. 0.024). Conclusion: Overexpression of miR-630 led induction of T-ALL cell growth and reduction of their apoptosis. These results emphasized that miR-630 contributed as an oncogenic microRNA in T-ALL cells

The metabolic syndrome is not associated with homocysteinemia: The Persian Gulf Healthy Heart Study

Background: It is uncertain whether homocysteine and the metabolic syndrome or its components are related in the general population, as studies investigating the association between homocysteine levels and insulin resistance have shown conflicting results. Methods: In an ancillary study to the Persian Gulf Healthy Heart Study, a cohort study of Iranian men and women aged ≥25 yr, a random sample of 1754 subjects were evaluated for the association of plasma homocysteine levels and the metabolic syndrome using National Cholesterol Education Program (NCEP)-Adult Treatment Panel (ATP)-III criteria. Total homocysteine levels and high sensitivity C-reactive protein (CRP) were determined by enzyme-linked immunosorbent assays. Results: Subjects with lower HDL-cholesterol and higher blood pressure showed significantly higher homocysteine levels (p=0.001 and p<0.0001; respectively). There was no significant difference in serum levels of homocysteine between subjects with and without the metabolic syndrome. In multiple logistic regression analysis, the metabolic syndrome did not show a significant association with serum homocysteine levels after adjusting for sex, age, smoking, fruit and vegetable intake pattern, body mass index, and physical inactivity. Concurrent elevated CRP levels and the metabolic syndrome also did not show a significant association with serum homocysteine levels after adjusting for sex, age, and lifestyle cardiovascular risk factors. Conclusions: There was no association between the metabolic syndrome using NCEP-ATPIII criteria and homocysteinemia in this study. These data refute the hypothesis that homocysteine levels are influenced by the metabolic syndrome, at least in general healthy population

A Pleurocidin-Like Peptide from Poecilia Mexicana Fish Induces Selective Cytotoxicity in Leukemia Jurkat Cells Through The Apoptosis Pathway

Objective: Some cationic anti-microbial peptides show a wide range of cytotoxic action versus malignant cells, which may lead to developing a novel group of antitumor medications. In the present study, the anticancer activity of pleurocidin-like peptide WF3 isoform X2 (AMP-WF3), from the Poecilia Mexicana fish, against leukemic cell line Jurkat was evaluated, and the cytotoxicity compared with the effects on normal cells, including peripheral blood mononuclear cells (PBMCs) and human dermal fibroblast (HDF) cells. Materials and Methods: In this experimental study, cells were treated with various dosages of AMP-WF3 for 24 hours. Using methyl thiazole tetrazolium salt reduction (MTT test), the effects of the AMP-WF3 on cell viability and toxicity were evaluated. The impact of this peptide on apoptotic pathways was examined using flow cytometry and Annexin V-PI stains. Additionally, the relative expression of the P53, P21, and BCL-2 genes was evaluated using a real-time polymerase chain reaction. Results: The Jurkat cell line was more susceptible to AMP-WF3 cytotoxicity [half-maximal inhibitory concentration (IC50)=50 μM], while normal cells (PBMCs and HDF) were less susceptible. Flow cytometry verified that the apoptotic activity of AMP-WF3 on Jurkat cells was significantly higher than that of HDF and PBMCs. Peptide-treated Jurkat cells were associated with increased expression of P21, and P53 genes. In contrast, the changes in P21, P53, and BCL-2 genes differed in PBMCs and HDF cells. In HDF cells, simultaneous increase of P21, P53, and BCL-2, and in PBMCs, only the increase of BCL-2 was observed. Conclusion: Our research showed that AMP-WF3 could be developed as a novel treatment agent with minimum side effects for ALL patients. © 2023 Royan Institute (ACECR). All rights reserved

Overexpression of MiR-506 in jurkat (Acute T cell leukemia) cell line

Background & Objective: Acute lymphoblastic leukemia (ALL) is a malignant disease that arises from various mutations in B or T-lymphoid progenitors. MicroRNAs (miRNAs) regulate gene expression by binding to the 3' untranslated region of proteincoding genes. Dysregulation of miRNA expression may result in the development of cancerous phenotypes. Therefore, for the first time in this field, the present study aims to investigate the effect of overexpression of miR-506 in Jurkat (acute T cell leukemia) cell line. Methods: In this study, Jurkat cell lines were cultured in RPMI-1640 medium. Next, miR-506 was transfected with concentrations of 50 and 100 nM with Lipofectamine 2000. The accuracy of the transfection was confirmed by the transfection of siRNA conjugated with FITC. 48 h after transfection, the cells were prepared for other tests (flow cytometry, MTT assay, and RNA extraction). The expression level of miR-506 in the cells was analyzed using the quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR). Finally, SPSS 21 software was used for the data analysis. Results: According to our results, the viability of cells in concentrations of 50 and 100 nM was significantly higher than the control group. By overexpression of miR-506, the expressions of pro-apoptotic genes (p53, p21) and anti-apoptotic gene B-cell lymphoma-2 (BCL-2) are decreased and increased, respectively. Conclusion: This study showed that miR-506 may function as an oncogenic miRNA in the T-ALL cell line. In conclusion, overexpression of miR-506 leads to an increase in viable cancer cells

Spectrum of mutations of thalassemia among couples from izeh city, khuzestan province, iran

Hemoglobinopathies are inherited blood disorders with an autosomal recessive pattern. We aimed to evaluate the frequency of mutations of thalassemia and hemoglobinopathies among couples referred to health centers of Izeh in Khuzestan Province, Iran. Methods: This cross-sectional study was performed on 150 couples referred to Izeh Health Centers in 2015-2018. DNA was isolated from peripheral venous blood samples and then the HBB gene was analyzed by using Sanger sequencing. For molecular analysis of α-globin gene, multiplex Gap-PCR and ARMS-PCR was performed to identify mutations of α-thalassemia. Results: DNA analysis revealed 13 different mutations for beta thalassemia in studied couples. Three mutations including 36/37 (-T), IVS-II-1 (G>A) and IVS-I-110 (G>A) accounted for 20.7, 19.3 and 13.3% of beta thalassemia mutations, respectively. For alpha thalassemia; α3.7 (49.5%),--MED (19.1 %) and-α4.2 (3.1%) were identified as the most common mutations. Conclusion: Considering common alpha and beta mutations of this geographic area of Iran could be useful concerning genetic counselling in of the population where the rate of consanguineous marriage is high

Simultaneous regulation of miR-451 and miR-191 led to erythroid fate decision of mouse embryonic stem cell

Objective(s): Various microRNAs (miRNAs) are expressed during development of mammalian cells, when they aid in modulating gene expression by mediating mRNA transcript cleavage and/or regulation of translation rate. miR-191 and miR-451 have been shown to be critical regulators of hematopoiesis and have important roles in the induction of erythroid fate decision. So, the aim of this study is investigation of the miR-191 and miR-451 roles in the controlling mouse embryonic stem cell (mESC) differentiation toward the erythroid lineage. Materials and Methods: mESCs were infected with either pCDH-miR-Off-191 viruses in pCDH-miR-Off-191 group or simultaneously with pCDH-miR-Off-191 and pCDH-miR-451 lentiviruses in simultaneous group. Then, the expression profiles of erythroid specific transcription factors and globin genes were analyzed using QRT-PCR on day 14 and 21 of differentiation. Flow cytometry analysis was used to evaluate of TER119 and CD235a erythroid specific surface markers. Results: Gata-1, Klf-1, Epor and globin chains were found to be expressed in pCDH-miR-Off-191 and in simultaneous groups. The majority of globin chains showed changes in their expression levels with progression of differentiation from day 14 to day 21. Flow cytometry results showed that miR-451 upregulation and miR-191 down-regulation is associated with the expression of TER119 and CD235a. Of these two groups analyzed, simultaneous group was most significantly potent in stimulation of erythroid fate decision of mESCs. Conclusion: Together, present data demonstrate that down-regulation of miR-191 alone can enhance the differentiation of mESCs. However, the simultaneous effect of miR-451up-regulation and miR-191 down-regulation is much stronger and can have more practical use in artificial blood production

Why so serious? Theorising playful model-driven group decision support with situated affectivity

This is the author accepted manuscript. The final version is available from Springer via the DOI in this record.An integrative approach to theorising behavioural, affective and cognitive processes in modeldriven group decision support (GDS) interventions is needed to gain insight into the (micro-)processes by which outcomes are accomplished. This paper proposes that the theoretical lens of situated affectivity, grounded in recent extensions of scaffolded mind models, is suitable to understand the performativity of affective micro-processes in model-driven GDS interventions. An illustrative vignette of a humorous micro-moment in a group decision workshop is presented to reveal the performativity of extended affective scaffolding processes for group decision development. The lens of situated affectivity constitutes a novel approach for the study of interventionist practice in the context of group decision making (and negotiation). An outlook with opportunities for future research is offered to facilitate an integrated approach to the study of cognitive-affective and behavioural micro-processes in model-driven GDS interventions.This work was supported in part by the EU FP7-ENERGY- SMARTCITIES-2012 (314277) project STEEP (Systems Thinking for Comprehensive City Efficient Energy Planning