Abstract. DNA methylation is a significant epigenetic modification which plays a key role in regulation of gene expression and influences functional changes in endometrial tissue. Aberrant DNA methylation changes result in deregulation of important apoptotic proteins during endometrial carcinogenesis and apoptosis resistance development. Evading apoptosis is still a major problem in the successful treatment of endometrial cancer patients. The aim of our study was to examine the promoter DNA methylation changes in 22 apoptosis-associated genes in endometrioid endometrial cancer patients, precancerous lesions and healthy tissue from various normal menstrual cycle phases using a unique pre-designed methylation platform. We observed as the first a significant difference in promoter DNA methylation status in genes: BCL2L11 (p < 0.001), CIDEB (p < 0.03) and GADD45A (p < 0.05) during endometrial carcinogenesis and BIK gene (p < 0.03) in different phases of normal menstrual cycle. The results of our study indicate that deregulation of mitochondrial apoptotic pathway can considerably contributes to the apoptosis resistance development and may be helpful in identifying of new potent biomarkers in endometrial cancer.
Since the molecular similarities and differences among physiological capacitation and cryocapacitation have not been studied in detail, this study was designed to assess the gene and protein expression levels of the Cation channel of sperm (CatSper) 1 and 2, sodium bicarbonate (Na+/HCO3−) cotransporter (NBC) and protein kinase A (PKA) in un-capacitated (control), in vitro capacitated (CAP) and cryopreserved (CRYO) bovine spermatozoa. All samples were subjected to motility evaluation using the computer assisted sperm analysis and chlortetracycline (CTC) assay for the assessment of the capacitation patterns. Furthermore, quantitative reverse transcription PCR (qRT-PCR) and Western blots were used to monitor the expression patterns of the selected capacitation markers. The results showed a significant reduction in the gene and protein expression levels of CatSper1 and 2 in the CRYO group when compared to the CAP group (p < 0.0001). In the case of NBC, the results were not significantly different or were inconclusive. While a non-significant down-regulation of PKA was found in the CRYO group, a significant reduction in the expression of the PKA protein was found in frozen-thawed spermatozoa in comparison to the CAP group (p < 0.05). In conclusion, we may hypothesize that while in vitro capacitated and cryopreserved spermatozoa exhibit CTC-patterns consistent with capacitation events, the molecular machinery underlying CTC-positivity may be different.
Summary Niobium, osmium, scandium, tungsten and vanadium are transition metals naturally occuring in the environment, particularly in the Earth’s crust. Anthropogenic activities, primarily industrial technologies, have precipitated significant alternations in the concentration and distribution of these metals. Such a dramatic change resulted, by all means, in the bigger potential of the environmental exposure, which poses a threat not only to humans but to all biological systems. Certain elements naturally occur in the animal and human plasma and tissues, but their concentrations are sometimes too low to be detected using the existing modern technologies. In small amounts, such elements are not harmful and some of them have even been suggested to have a beneficial role in the human or animal physiology. However, exposure to excessive antropogenically elevated levels can exert serious negative effects on the environment, agriculture and health. The findings summarized in this paper provide a review of the current knowledge about the implications of the transition metals considered on the health, accentuating the insufficiency and need for more relevant data.
The aim of this study was to assess the dose- and time-dependent in vitro effects of ferrous sulphate (FeSO4.7H2O) on the motility parameters, viability, structural and functional activity of bovine spermatozoa. Spermatozoa motility parameters were determined after exposure to concentrations (3.90, 7.80, 15.60, 31.20, 62.50, 125, 250, 500 and 1000 μM) of FeSO4.7H2O using the SpermVisionTM CASA (Computer Assisted Semen Analyzer) system in different time periods. Cell viability was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide (MTT) assay, and the Annexin V-Fluos was applied to detect the membrane integrity of spermatozoa. The initial spermatozoa motility showed increased average values at all experimental concentrations compared to the control group (culture medium without FeSO4.7H2O). After 2 h, FeSO4.7H2O stimulated the overall percentage of spermatozoa motility at the concentrations of ≤ 125 μM. However, experimental administration of 250 μM of FeSO4.7H2O significantly (P < 0.001) decreased the spermatozoa motility but had no negative effect on the cell viability (P < 0.05) (Time 2 h). The lowest viability was noted after the addition of ≥ 500 μM of FeSO4.7H2O (P < 0.001). The concentrations of ≤ 62.50 μM of FeSO4.7H2O markedly stimulated (P < 0.001) spermatozoa activity after 24 h of exposure, while at high concentrations of ≥ 500 μM of FeSO4.7H2O the overall percentage of spermatozoa motility was significantly inhibited (P < 0.001) and it elicited cytotoxic action. Fluorescence analysis confirmed that spermatozoa incubated with higher concentrations (≥ 500 μM) of FeSO4.7H2O displayed apoptotic changes, as detected in head membrane (acrosomal part) and mitochondrial portion of spermatozoa. Moreover, the highest concentration and the longest time of exposure (1000 μM of FeSO4.7H2O; Time 6 h) induced even necrotic alterations to spermatozoa. These results suggest that high concentrations of FeSO4.7H2O are able to induce toxic effects on the structure and function of spermatozoa, while low concentrations may have the positive effect on the fertilization potential of spermatozoa.
Licensed under a Creative Commons Attribution 4.0 International License Peach is a popular sweet fruit with a very good climate adaptation and high production. It offers many health benefits determined by its biochemical composition. However, sensitive people can be sensitised by Pru p 3 non-specific lipid transfer protein family directly or through cross reaction as a member of Bet v 1 homologues. The majority of research is focused on the protein, less data can be found in genomics or transcriptomics. This study performed RFLP analysis by chosen restricted enzymes (BfaI, MseI, NlaIII) for non-coding region of Pru p 3 (NCBI: KC311811.1) as a tool to distinguish closely related isoforms of the allergen. BfaI cut amplicon into 5 fragments corresponding to the Yulu variety in silico cleavage and polymorphism was not detected. For MseI and NlaIII polymorphisms were found in the cleavage sites, two types of restriction profiles were created for both. None of the NlaIII profiles corresponds to the restriction profile of in silico cleavage. The study confirms the varietal differences in Pru p 3 gene and supports a hypothesis that allergenicity depends on both qualitative and quantitative factors that are different and specific to each variety.
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