Purpose The length of GT-repeats polymorphic region in the promoter of human Heme oxygenase-1 gene (HO-1) alters the level of its transcriptional activity in response to oxidative stresses. Decreased level of HO-1 protein in the seminal plasma has been reported to be associated with oligospermia and azoospermia in male infertility. This is the first study to investigate the association between GT-repeats expansion in the promoter of the HO-1 gene and male infertility. Methods The frequencies of different GT-repeats alleles in the promoter of HO-1 gene were determined in 100 cases and 100 normal controls using PCR-PAGE, ABI fragment analysis genotyping and sequencing analysis.Results All alleles were classified into S and L alleles. S alleles were specified as number 0 to 3 with <27 GT-repeats and L alleles were specified as number 4 to 6 with >27 repeats. The L allele frequency was significantly higher among case group (54.5%) than that was obtained in the normal control group (37.5%). Statistical analysis provided a significant relationship between L allele and male infertility (P<0.001). Conclusions This study shows for the first time that GTrepeats expansion in promoter of the HO-1 gene is associated with oligospermia and azoospermia among Iranian infertile cases.
Background:The genome of Enterococcus has a large number of repetitive sequences that are randomly distributed over DNA. In ERIC-PCR, a separate pattern is obtained for each strain and is considered a separate type. Prokaryotic and eukaryotic genomes include dispersed repeat sequences that are relatively short non-coding, and dispersed in the bacterial genome. BOX-PCR and ERIC-PCR primers are complementary to these repeat sequences and allow for dedicated binding and unique BOX-PCR fingerprint patterns and ERIC-PCR with reproducibility capability.Methods: In this cross-sectional descriptive study, based on previous studies and 95% confidence level using the formula n = z2P (1-P)/d2 and acceptable error 0.05, total of 60 Enterococcal Faecalis strains were cultured from sterile specimens on KF agar medium and incubated at 37°C for 24h and were identified by biochemical tests of suspected Enterococcal Faecalis. DNA samples were extracted and BOX-PCR and ERIC-PCR tests were performed.Results: All strains were distinguished in 25 distinct clusters at the level of similarity of 58%. Also, by analyzing the ERIC-PCR results, all strains were segregated into 15 separate clusters at a similar level of 58%.Conclusion: Molecular fingerprinting with BOX-PCR has a better subtraction and differentiation than ERIC-PCR for typing bacterial isolates and is widely used in epidemiological studies and trace source of infection and taxonomy.
Background: MicroRNAs (miRNAs), short regulatory RNAs, function as negative regulators able to modulate gene expression. Just as other genetic variant, single nucleotide polymorphisms (SNPs) in miRNA genes, may have an impact on their expression and/or maturation and hence leading to different consequences in carcinogenesis. Accordingly, this study aimed to assess the frequency of miR-146a G/C ( rs2910164 ) polymorphism and its association with susceptibility to breast cancer in Iranian women. Methods: We conducted a case-control study using Tetra ARMS polymerase chain reaction (Tetra ARMS PCR) method in 100 Iranian female participants (50 breast cancer patients and 50 controls). Besides, a number of sequenced samples were chosen to confirm the accuracy of genotyping by Tetra ARMA PCR. SPSS software was utilized for all statistical analyses. The odds ratios (ORs) and 95% confidence intervals (95% CIs) were applied to analyze the association between the SNP frequency and breast cancer. Results: The frequency of genotypes for G/G, G/C, and C/C were 23 (46%), 26 (52%), and 1 (2%) among cases and 15 (30%), 33 (66%), and 2(4%) among controls, respectively. The results generated by the groups did not show any significant correlation between miR-146a G/C ( rs2910164 ) polymorphism and breast cancer, either at genotype or allele levels (P>0.05). F-SNP-based in silico analysis indicated possible modifications in transcriptional regulations induced by miR-146a G/C (rs2910164) variations. Conclusion: Overall, our results indicated no correlation between miR-146a G/C (rs2910164) polymorphism and genetic susceptibility to breast cancer in Iranian female populations. However, these findings need to be further confirmed by analyses of a larger number of cases.
Background: It is estimated that 1-5% of couples suffer from recurrent pregnancy loss (RPL). Recent studies have shown the effects of gene polymorphisms in RPL. Objective: The aim of this study was to evaluate 3 gene polymorphisms including rs1048943 of CYP1A1, rs28371725 of CYP2D6, and rs7830 of NOS3 in idiopathic RPL to identify their association with RPL. Materials and Methods: Blood samples were collected from 136 women with at least 2 consecutive idiopathic miscarriages (case group) and 136 women with no history of miscarriage and at least one successful pregnancy (control group) from the Iranian Azeri population. This study was carried out between April 2018-April 2020. Amplification-refractory mutation system polymerase chain reaction was used for the rs7830, rs1048943 and rs28371725 polymorphisms in order to genotype each extracted genomic DNA sample. After that, Chi-square, Fisher’s exact test and logistic regression were used to investigate whether each of these polymorphisms is associated with RPL. Results: Among these polymorphisms, only rs1048943 of CYP1A1 showed a statistically significant association with RPL in the Iranian Azeri women studied. Conclusion: Our results suggest that CYP1A1 gene polymorphisms might be associated with a reduced risk of RPL. Further studies in other populations and in the same population with a larger sample size, as well as functional genomics analyses such as gene expression analyses or epigenetic studies are required to validate our results. Key words: Recurrent pregnancy loss, Polymorphism, CYP1A1, CYP2D6, NOS3.
Background: Temperature changes cause testicular dysfunction. It has been observed that testicular hyperthermia leads to oxidative stress and as a result a severe reduction in testicular parameters. Causes a severe reduction in Sperm parameters to become oxidative due to stress. Recently, natural plant materials and magnetic nanoparticles have been considered. In the internal mitochondrial apoptosis pathway, gen bcl2 is a target of miR455. Objectives: The present study aimed to investigate the effects of titanium dioxide nanoparticles and improve their impacts by using the antioxidant curcumin on sperm parameters by investigating changes in expression miR455 in response to temperature-induced stress in scrotal hyperthermia rats. Methods: After preparation, the rats were placed in a hot water bath at 43°C. for 30 minutes for six consecutive days. The rats were then divided into eight groups. We used TiO2 nanoparticles at a concentration of 0.03 mg/kg of body weight and curcumin at a concentration of 0.02 mg/kg of body weight. After killing the animals, such parameters of sperm as viability, concentration, motility, and morphology of spermatozoa were studied. RNA extraction and cDNA synthesis were performed using appropriate kits. A gene primer was designed and RT-PCR was used to assess gene expression. The t-test and ANOVA were used to examine differences between different groups. Data analysis was performed using Prism8 software and SPSS version 26. Results: The results showed that miR455 expression was lower in the treatment groups and was associated with curcumin (P < 0.05). A positive effect of curcumin on improving sperm parameters in rats with scrotum hyperthermia and a negative and toxic effect of TiO2 nanoparticles were shown. However, a significant improvement in sperm parameters was observed when rats were given TiO2 nanoparticles along with curcumin. Conclusions: The changes in the expression miR455, shown in curcumin have controlled the damage to TiO2 nanoparticles. It seems that miRNA455 can be used as a marker to predict sperm health status. So Curcumin can play a protective role in reducing the toxic effects of testicular hyperthermia as well as titanium dioxide nanoparticles.
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