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Background: The expression of forkhead box O (FOXO) was found to be connected with developing rheumatoid arthritis (RA), an inflammatory autoimmune disorder. The current study is intended to assess the expression and methylation status of the FOXO1 gene in individuals with recently diagnosed RA, before and after the administration of customary disease-modifying antirheumatic drugs (DMARDs). Methods: Twenty participants were investigated in this study. The assessment of the FOXO1 gene expression in peripheral blood was done by real-time PCR, and the status of FOXO1 promoter methylation was ascertained via quantitative methylation-specific PCR (Q-MSP) before and after the administration of DMARDs for six months. Results: Following DMARDs treatment, the study discovered a decrease in FOXO1 gene expression. However, the decline did not meet the criteria for statistically meaningful (P = 0.087). The expression of the FOXO1 gene was positively correlated with RA disease activity pre- and post-treatment with DMARDs (P = 0.009, r = 0.567 and P = 0.001, r = 0.656, respectively). Moreover, the study showed no alterations in the amount of DNA methylation of the FOXO1 promoter in newly diagnosed RA patients who had not yet received DMARDs, as compared to DMARDs-treated RA patients. Conclusion: Altogether, this study suggests that DMARDs treatment may reduce FOXO1 gene expression, potentially helping to alleviate the pro-inflammatory effects associated with this gene. [ABSTRACT FROM AUTHOR]

Background: Infertility affects about 15% of couples during reproductive age and male factor infertility accounts for half of these cases. This study aimed to evaluate the expression of leptin, KISS1, leptin receptor, and HOTAIR genes in blood and semen samples of individuals diagnosed with oligospermia in comparison to healthy controls. Methods: In the current investigation, we studied 36 semen and 30 blood samples from fertile oligospermic men as well as the same number of healthy controls. RNA was extracted and cDNA was synthesized. Real-time polymerase chain reaction (PCR) was conducted to assess the gene expression levels. Statistical analysis was performed using Graph Pad Prism software. The results were reported as mean±SEM and any P< 0.05 were considered statistically significant. Results: A significant increase in the expression of the leptin gene in infertile males, particularly in semen samples (P< 0.001) was found when compared to the healthy controls. On the other hand, significant decrease in the expression of the KISS1 (P< 0.0001) and HOTAIR (P< 0.001) genes, particularly in semen samples were found when compared to the controls. As it was shown, the expression of leptin receptor had no significant effects on male infertility. Additionally, there may be a potential correlation between the expression of HOTAIR and the leptin and KISS1 genes, with a negative correlation observed between HOTAIR and leptin, and a positive correlation between HOTAIR and KISS1 in both blood and semen samples. However, further investigations are necessary to establish the statistical significance of these correlations. Conclusion: According to the results, leptin, KISS1, and HOTAIR genes seem to be affected in oligospermia, however, further studies with higher sample sizes are necessary. [ABSTRACT FROM AUTHOR]

Background: Extensive use of different nanoparticles caused significant concerns about their biological safety. Objective: This study aimed to evaluate the effects of cryopreservation on ram semen after adding magnetic nanoparticles (MNPs) to separate X and Y chromosome‐bearing spermatozoa. Methods: The experimental ram sperms in this research included treated spermatozoa (50 μg/ml MNPs) and non‐treated spermatozoa. DNA damage of spermatozoa was examined using an acridine orange (AO) assay. Sperm viability, membrane functionality, abnormality and malondialdehyde (MDA) level were also measured. Results: Results indicated that the pre‐treatment of ram semen extender with MNPs did not significantly affect the semen parameters such as viability, membrane functionality, abnormality, as well as lipid peroxidation (LPO) levels and DNA integrity in comparison with the control group (p < 0.05). Conclusions: These observations suggest that pre‐treatment of ram semen extender with MNPs after semen sexing did not have adverse effects on different semen parameters after cryopreservation. In the current study, we evaluated the effects of cryopreservation on ram semen after adding magnetic nanoparticles (MNPs), then after thawing some semen parameters were evaluated. [ABSTRACT FROM AUTHOR]

Pre‐conceptual sex selection is still a highly debatable process whereby X and Y chromosome bearing spermatozoa are isolated before oocyte fertilization. Recently, magnetic nanoparticles (MNP) have been used to determine X and Y chromosomes bearing spermatozoa as a result of searching for a cheap, highly efficient method using non‐toxic materials. This study aimed to recover the sperm bearing X chromosomes in ram with different concentrations of MNP and then evaluate the success of this method using polymerase chain reaction (PCR). Ram sperms were divided into four groups, treated with 0 (control), 50, 100 and 200 μg/ml MNP, respectively. MNP was used to restore sperm cells bearing X chromosomes. Upon recovery, the PCR was performed to identify the X and Y sperms, Methyl ThiazoleTetrazolium (MTT), to assess MNP toxicity and sperm viability and acridine orange (AO) to evaluate sperm DNA integrity. The results of PCR revealed that the treatment of spermatozoa‐ bearing X chromosomes with 50 μg/ml MNP had the highest effects on the recovery of X sperm rather than the other concentrations of MNP. However, the concentrations of MNP did not have any toxic effects on spermatozoa, sperm viability and, DNA integrity, but the high concentration of MNP (200 μg/ml) significantly reduced DNA integrity. According to MTT and AO results, the concentrations of MNP used in this study had no toxic effects on spermatozoa and did not reduce the sperm viability and DNA integrity, except that 200 μg/ml MNP significantly reduced DNA integrity. [ABSTRACT FROM AUTHOR]

Background: Sex selection of sperm by separating X- and Y-chromosome bearing spermatozoa is critical for efficiently obtaining the desired sex of animal offspring in the livestock industry. The purpose of this study was to produce a goat polyclonal antibody (pAb) against the bovine Sex Determining Region Y chromosome (bSRY) to separate female- and male-bearing spermatozoa. Methods: To produce a goat polyclonal antibody against bSRY, a female goat was subcutaneously immunized with 27 kDa of recombinant bSRY (rbSRY) protein as the antigen. The anti-bSRY pAb was purified by ion-exchange chromatography. The purity of the pAb was determined using the SDS-PAGE method. The biological activity of the anti-bSRY pAb was examined using PCR to assess the binding affinity of pAb for the bSRY antigen and commercially sexed bull sperm. Results: The total amount of purified anti-bSRY pAb was approximately 650 mg/goat serum (13 mg/mL). Interestingly, our data showed that the binding affinity of our pAb to the Y bearing was high, while the binding affinity of that to the X-chromosome bearing sperm was similar to the negative control. Conclusions: In conclusion, our findings show that the goat anti-SRY pAb specifically binds to Ychromosome bearing sperm that suggesting its potential use for sex selection. [ABSTRACT FROM AUTHOR]

Glycated proteins are generated by binding of glucose to the proteins in blood stream through a nonenzymatic reaction. Hemoglobin A1c (HbA1c) is a glycated protein with glucose at the N‐terminal of β‐chain. HbA1c is extensively used as an indicator for assessing the blood glucose concentration in diabetes patients. There are different conventional clinical methods for the detection of HbA1c. However, enzymatic detection method has newly obtained great attention for its high precision and cost‐effectiveness. Today, fructosyl peptide oxidase (FPOX) plays a key role in the enzymatic measurement of HbA1c, and different companies have marketed HbA1c assay systems based on FPOX. Recent investigations show that FPOX could be used in assaying HbA1 without requiring HbA1c primary digestion. It could also be applied as a biosensor for HbA1c detection. In this review, we have discussed the recent improvements of FPOX properties, different methods of FPOX purification, solubility, and immobilization, and also the use of FPOX in HbA1c biosensors. [ABSTRACT FROM AUTHOR]

Sex determining region Y gene (SRY) is located on Y chromosome and encodes a protein with 229 amino acids. In this study, ORF region of SRY with a length of 690 bp was synthesized using PCR and ligated to pET28a (+), then transformed in E.coli DH5α. E.coli BL21 (DE3) strain was chosen to express recombinant bovine SRY protein. A set of optimization steps was taken including different concentrations of IPTG, glucose, and temperatures at differed incubation times after the induction. Results showed that temperature points and different concentrations of IPTG and glucose had a significant effect (p < 0.01) on total protein and recombinant bovine SRY. After purification, various temperatures and concentrations of IPTG showed meaningful effects (p < 0.01) on the solubility of expressed recombinant SRY. Highest soluble rSRY protein amount was achieved where 0.5 mM IPTG and 0.5% glucose was used at 20°C during induction. In the absence of glucose, the highest amount of soluble recombinant SRY levels were achieved at the concentrations of 0.8 mM of IPTG at 28°C, 20°C, and 1.5 mM IPTG at 37°C during induction for 16, 24, and 8 hours, respectively. Regarding the results obtained in this study, it could be stated that by decreasing temperature and inducer concentration, soluble bovine SRY protein expression increases. [ABSTRACT FROM AUTHOR]

Objective: Integrin molecules and chemokines play critical role in Lymphocytes and leukocytes migration and recirculation during autoimmune disorders. Normally gut-homing T cells express CCR9 and integrin β7 in order to home back to lamina propria of intestine from peripheral blood. Our study was conducted to evaluate expression levels of the chemokine receptor CCR9 and the integrin β7 in circulating leukocytes and lymphocytes from newly diagnosed Rheumatoid arthritis (RA) patients and compare these values with the same data obtained from healthy individuals without inflammatory disease. Material and Methods: Total RNA was extracted from peripheral blood samples immediately cDNA was synthesized and finally gene expression of CCR9 and β7 evaluated by quantitative Real time PCR. The T-test and ANOVA were used in order to compare the differences between two groups. Results: The β7 gene expression showed significant difference between RA patients and healthy controls (P value= 0.007). Conclusion: In our newly diagnosed patients who did not received any medication for RA the gene expression of integrin β7 was higher than healthy controls. CCR9 gene expression did not reach significant different between two groups. Our results bring up disposability that over expression β7 gene may have a role in pathogenesis of RA. [ABSTRACT FROM AUTHOR]

Background: Inclusion body formation in E. coli is a significant problem in recombinant protein production.The aim of this study was to improve the solubility of recombinant bovine sex determining region Y protein(SRY) in BL21 (DE3) E. coli cells., Methods: In this research two recombinant bovine SRY (rbSRY) sequences were analyzed; these were wildtype SRY (wtbSRY) and codon-optimized SRY (cobSRY). Their expression in various culture conditions was examined; these differences included IPTG concentrations, temperatures, and media stabilizers., Results: IPTG and temperature significantly affected rbSRY solubility (P < 0.001). The optimum IPTG concentration and temperatures for wtbSRY and cobSRY induction were 0.3 mM at 27 and 32 °C,respectively. In addition, arginine and sorbitol concentrations significantly affected rbSRY solubility (P <0.01). Solubility of rbSRY protein was highest from the cobSRY construct in the presence 0.2 M arginine and 0.3 M sorbitol. The highest inclusion body production occurred with high glucose concentrations., Conclusion: We found that modifications in temperature and IPTG and stabilizer concentrations affected rbSRY solubility.