Pathobiology Research
Year:2017 | Volume:20 | Issue:1 |Papers Count:7

Objective: Autophagy (self-digestion) is a highly regulated process for the degradation of damaged proteins and intracellular components. Autophagy has multifunctional roles in the protection of cellular homeostasis. Beclin1 is a key regulator molecule in autophagosome formation. Inhibition of autophagy by destruction of the Beclin 1 allele creates sensitivity to metabolic stress. Inhibition of the autophagy under conditions of nutrient deprivation in tumors resistant to apoptosis can lead to necrosis, inflammation and increased tumor growth. This study aims to assess the effect of autophagy induction on the necrosis pathway of MDCK cells.Methods: We evaluated induction of autophagy by the Beclin1 gene in MDCK cells and assessed the percentage of necrosis cell death by flow cytometry using an Annexin V Staining kit. In order to induce autophagy, the recombinant pcDNA3.1-Beclin 1 was transfected into the MDCK cell line using lipofectamine TM 3000.Results: Overexpression of the Beclin1 gene in MDCK cells led to induction of autophagy as seen by intracellular autophagosomal indicator LC3-II staining. There were 9.92% positive LC3 structures in transfected cells and 0.15% in untransfected cells. In the transfected and control groups, the rate of necrosis cell death was 1.66% and 0.06%, respectively.Conclusion: Crosstalk between autophagy and necrosis pathways might affect the fate of thecell life span. Strategies that involve in modulation of autophagy and cell death might lead to therapeutic interventions in diseases. Therefore manipulation of cell death pathways could create new areas in therapeutic uses and interventions.

Objective: Considering the high incidence of acute lymphoblastic leukemia (ALL), it is necessary to research this illness and determine genetic markers associated with it. The role of DNMT I gene expression during treatment with mercaptopurine drugs in children with B-cell ALL (B-ALL) is important because its changes reflect the effectiveness or ineffectiveness of the drugs. We have investigated the effect of mercaptopurine drugs on DNMT I expression in children with B-ALL.Methods: This analytical study assessed 8 B-ALL children who referred to Tehran Children's Medical Center and 10 healthy children referred to the Yazd Central Laboratory by convenience selection in 2016. Blood samples were obtained before and after treatment, and extraction of total RNA from each sample was performed for realtime qPCR of the target gene (DNMT I). Simultaneously, we evaluated GAPDH, a housekeeping gene. Data from gene expressions were compared by the paired t-test, using SPSS-16 software.Results: DNMT I gene expression was significantly decreased after mercaptopurine administration in children with B-ALL (P<0.02).Conclusion: Our findings suggest that the mercaptopurine drug may affect DNMT I gene regulation. This epigenetic effect may explain the mechanism of drug action, possibly serve as a diagnostic factor, and a means of better monitoring for patients with ALL.

Objective: This experimental study evaluatedthe effect of ovarian hormones on the expression of genes related to implantation in human endometrial stromal cells in vitro.Methods: After confirmation ofnormal endometrial tissue collected from hysteroscopy according to hematoxylin and eosin staining, we isolated the endometrial cells with collagenase type 1. The cells were passed through 100 and 40 μm filters. We cultured the collected stromal cells in DMEM/F-12 medium to the fourth passage. Passage-4 cells were examined for 7 days in two groups - control (without hormones) and experimental groups with different concentrations of estrogen (E2; 0.3, 0.7 and 1 nmol/L) and a constant concentration of progesterone (P4; 63.5 nmol/L). After culture, the cell viability, proliferation and the expression of genes related to implantation, αv and β3 integrins, leukemia inhibitory factor receptor (LIFR), and interleukin-1 receptor (IL-1R) were assessed by staining with propidium iodide and acridine orange, the MTT assay, and real time RT-PCR.The data were analyzed using SPSS software.Results: There were 99.9% viable cells in all groups. Cell proliferation increased significantly in the 0.3 nmol/L treated group compared with the control group and the group treated with 0.7nmol/L (P£0.04). However, the group treated with 1 nmol/L did not significantly change compared to the other groups. In contrast to IL1-R and LIFR, expressions of the av and b3 integrins significantly increased in the 0.3 nmol/L treated group compared to the control (P£0.02).Conclusion: The combined use of estrogen and progesterone in an endometrial stromal cell culture had an effect on the proliferation and expression of some genes related to implantation.

Objective: Familial hypercholesterolemia (FH) is an autosomal dominant disease mainly attributed to mutations in the low-density lipoprotein receptor gene (LDLR). This study aims to investigate molecular changes in the LDLR gene in patients with high cholesterol in individuals from Ardebil Province, Iran.Methods: We evaluated 100 patients with suspected FH from Ardebil Province. DNA samples using primers LDLR gene and exon 10 PCR-SSCP method was tested and modified bands on gel electrophoresis detected and subsequently examined by DNA sequencing.Results: We evaluated 100 patients with suspected FH, 43 males and 66 females. The average age of 50.68 and 281.81 had average cholesterol levels of the subjects. In this study, we identified a polymorphism 1413G>A LDLR gene. Allele G, 70% of the population studied and the A allele is 30% of the subjects to be included.Conclusion: The findings of this study showed that polymorphism of the LDLR gene in FH 1413GA was not the main role, but could indirectly affect, and possibly other exons of the gene or other genes in the development of FH in the region have.

Objective: Trichomonas vaginalis (T. vaginalis) is a pathogenic protozoan of human reproductive-urinary systems that causes trichomoniasis. The disease is the most important non-viral sexually transmitted infection worldwide. Various laboratory methods have been used to diagnoseT. vaginalis. Based on the actin gene, 6 genotypes (H, G, E, I, M, N) of T. vaginalishave been identified. In most studies, the clinical samples were cultured initially and then genotyped. In this study, we sought to identify and determine the genotype of T. vaginalisin urine samples from infected women in Mahshahr, Khuzestan Province, Iran.Methods: Urine samples were collected from 2200 women who referred to the Laboratory of Imam Musa Kazim Hospital of Mahshahr. After microscopical examination, we extracted the parasite’s DNA from 34 positive urine samples. Then, the actin gene of the parasite was amplified by nested-PCR. Finally the PCR products of actin gene were sequenced.Results: Totally, 34 samples (54.1%) tested positive for T. vaginalis. After sequencing, the genotype of the parasite was identified as E in Mahshahr.Conclusion: Genotype E of T. vaginalis is the single genotype among women residents of Mahshahr. No genotypic variation was seen.

Objective: Elderly age is accompanied with reduced muscle strength and mass. Vascular endothelial growth factor (VEGF) is the most important stimulant of angiogenesis in muscles which, by increasing blood flow to the muscles, causes muscle strength and endurance. Exercise training leads to increased nitric oxide (NO) and release of VEGF by raising the shear stress during proliferation of endothelial cells and the process of angiogenesis. Garlic also modulates blood flow. This study has assessed the effects of regular swimming training combined with the consumption of garlic extract on the levels of NO plasma and VEGF tissue of soleus slow-twitch and gastrocnemius fast-twitch muscles in aged rats.Methods: Male aged rats (40-50 weeks) with an average weight of 250-300 g were randomly divided into 5 groups (n=7 rats per group) - control, saline, aerobic training, garlic, and aerobic training+garlic. The swimming training program was scheduled for 50 m daily, 3 times per week for 8 weeks. The groups that received supplements and supplementary training were given a daily dose by gavage of 1 ml/kg body weight of the garlic extract for 8 weeks. We collected tissue and blood samples 48 h after the last training session and after a 10-12 h fasting period. The amount of NO and VEGF were detected by colorimetry and ELISA, respectively.Results: One-way ANOVA revealed that regular aerobic training increased plasma NO (P=0.001), VEGF of the soleus muscle tissue (P<0.001), and VEGF of the gastrocnemius muscle (P=0.004) in the aged rats. In the group that received garlic extract, there was a significant increase in NO levels (P=0.001), and VEGF in the soleus muscle tissue (P=0.007) and gastrocnemius muscle tissue (P=0.015). Combined training and garlic supplement significantly increased the plasma level of NO (P<0.001), and VEGF in the soleus muscle tissue (P<0.001) and gastrocnemius muscle tissue (P=0.001) compared to the control and saline groups.Conclusion: Consumption of garlic extract alone and combined with aerobic training significantly increased plasma NO levels, and VEGF of the soleus and gastrocnemius muscle tissues. However, while the extents of increase in the training combined with garlic extract group was higher than that of the garlic group, there was no significant difference observed between the two groups. The lack of significant difference between these two groups might be due to the intensity and type of the training and would need additional research.