Evaluation of Sperm Parameters and DNA Integrity Following Different Incubation Times in PVP Medium.

PURPOSE: Polyvinylpyrrolidone (PVP) is a chemical material used in intracytoplasmic sperm injection (ICSI) program.  The aim of this study was to investigate the ideal time that sperm can be safely incubated in PVP with less structure and DNA damage. METHOD: Thirty-one Oligoasthenoteratospermia (OAT) samples were used. Sperm samples were prepared by discontinuous density-gradients method and incubated in 10% PVP at different time intervals (0, 5, 10, 15, 20, and 30 min). The effect of PVP was assessed on sperm DNA fragmentation and viability via SCD assay and Eosin-nigrosin staining respectively. RESULTS: Data showed there was a significant increase in sperm DNA fragmentation at 10 min compared to 0 min. The viability rate also significantly reduced at 10 min  compared to 0 min. CONCLUSION: As a result, sperm samples could be incubated with PVP for less than 10 min.  While prolonged incubation may significantly damage the sperm DNA integrity and viability.

Is there any relationship between human sperm parameters and protamine deficiency in different groups of infertile men? / ¿Existe alguna relación entre los parámetros del esperma humano y la deficiencia de protamina en varios grupos de varones infértiles?

OBJECTIVE: Abnormality in Histone-Protamine replacements has been indicated to cause sperm DNA damage and infertility. The aim of the present study was to investigate the relationships between sperm parameters in oligospermia, asthenospermia, and teratospermia with protamine deficiency in infertile men. MATERIAL AND METHOD: In this case-control study, we had three experimental groups including oligospermia (n=100), asthenospermia (n=100), and teratospermia (n=100) as well as normospermia (n=100) as controls. Sperm analyses were performed according to the recommendations of the World Health Organization (WHO, 2010) and sperm chromatin quality was assessed using Chromomycin A3 (CMA3) staining for each sample. RESULTS: The comparison of the data between groups indicated that the percentage of spermatozoa with protamine deficiency was significantly different in patients with oligospermia, asthenospermia, and teratospermia when compared with control ones. However, there was no significant correlation between sperm nuclear protamine deficiency and their parameters of the men with teratospermia using CMA3 test. Regarding the oligospermia and asthenospermia semen samples, the findings showed the negative correlations between the sperm nuclear protamine deficiency and progressive motility as well as immobility (p < 0.001). CONCLUSION: The higher proportion of spermatozoa with abnormal chromatin packaging was observed in asthenospermic samples than those from other experimental groups as well as controls. It seems that normal morphology cannot have a valuable predictive value for good chromatin quality of spermatozoa, as much as normal motility characteristics, since samples with high mobility rates often have lower protamine deficiencies. The findings may provide a supportable promoting the future wider clinical application of chromatin/DNA integrity testing along with the semen analysis in male infertility

OBJETIVO: Se ha indicado que la irregularidad en los reemplazos de histona-protamina provoca daño en el ADN del esperma e infertilidad. El objetivo del presente estudio fue investigar las relaciones entre los parámetros espermáticos en oligospermia, astenospermia y teratospermia con deficiencia de protamina en varones infértiles. MATERIAL Y MÉTODO: En este estudio de casos y controles, hubo 3 grupos experimentales que incluían oligospermia (n=100), astenospermia (n=100) y teratospermia (n=100), así como normospermia (n=100) como controles. Los análisis de esperma se realizaron de acuerdo con las recomendaciones de la Organización Mundial de la Salud (OMS, 2010), y se evaluó la calidad de la cromatina de los espermatozoides utilizando la tinción con Chromomycin A3 (CMA3) para cada muestra. RESULTADOS: La comparación de los datos entre los grupos indicó que el porcentaje de espermatozoides con deficiencia de protamina fue considerablemente diferente en pacientes con oligospermia, astenospermia y teratospermia en comparación con la de los controles. Sin embargo, no hubo una correlación importante entre la deficiencia de protamina nuclear de esperma y sus parámetros de los varones con teratospermia cuando se utilizaba la prueba de CMA3. En cuanto a las muestras de semen de oligospermia y astenospermia, los hallazgos mostraron las correlaciones negativas entre la deficiencia de protamina nuclear de esperma y la movilidad progresiva, así como la inmovilidad (p < 0,001). CONCLUSIÓN: La mayor proporción de espermatozoides con un empaquetado de cromatina anómalo se observó en las muestras astenospérmicas que en las de otros grupos experimentales, así como en los controles. Parece que la morfología normal no puede tener un valor diagnóstico valioso de la buena calidad de la cromatina de los espermatozoides, tanto como las características normales de movilidad, ya que las muestras con altas tasas de movilidad a menudo tienen menores deficiencias de protamina. Los hallazgos pueden ofrecer un soporte que promueva la futura aplicación clínica más amplia de las pruebas de integridad de la cromatina/ADN junto con el análisis del semen en la infertilidad masculina

Is there any relationship between human sperm parameters and protamine deficiency in different groups of infertile men?

OBJECTIVE: Abnormality in Histone-Protamine replacements has been indicated to cause sperm DNA damage and infertility. The aim of the present study was to investigate the relationships between sperm parameters in oligospermia, asthenospermia, and teratospermia with protamine deficiency in infertile men. MATERIAL AND METHOD: In this case-control study, we had three experimental groups including oligospermia (n=100), asthenospermia (n=100), and teratospermia (n=100) as well as normospermia (n=100) as controls. Sperm analyses were performed according to the recommendations of the World Health Organization (WHO, 2010) and sperm chromatin quality was assessed using Chromomycin A3 (CMA3) staining for each sample. RESULTS: The comparison of the data between groups indicated that the percentage of spermatozoa with protamine deficiency was significantly different in patients with oligospermia, asthenospermia, and teratospermia when compared with control ones. However, there was no significant correlation between sperm nuclear protamine deficiency and their parameters of the men with teratospermia using CMA3 test. Regarding the oligospermia and asthenospermia semen samples, the findings showed the negative correlations between the sperm nuclear protamine deficiency and progressive motility as well as immobility (p<0.001). CONCLUSION: The higher proportion of spermatozoa with abnormal chromatin packaging was observed in asthenospermic samples than those from other experimental groups as well as controls. It seems that normal morphology cannot have a valuable predictive value for good chromatin quality of spermatozoa, as much as normal motility characteristics, since samples with high mobility rates often have lower protamine deficiencies. The findings may provide a supportable promoting the future wider clinical application of chromatin/DNA integrity testing along with the semen analysis in male infertility.

Association of IL-17 and IL-23 follicular fluid concentrations and gene expression profile in cumulus cells from infertile women at risk for ovarian hyperstimulation syndrome.

This study determined the association between the levels of interleukin (IL)-17 and IL-23 in follicular fluid (FF), as well as their mRNA levels in cumulus cells from infertile women at risk for ovarian hyperstimulation syndrome (OHSS). In this case-controlled study, the control group (n = 40) was infertile women whose partners had male factor infertility, whereas the case group (n = 40) was infertile women at risk of OHSS. IL-17 and IL-23 concentrations in FF were measured using an enzyme-linked immunosorbent assay method, whereas the mRNA expression levels of IL-17 and IL-23 of cumulus cells were determined using RT-PCR. Significantly higher levels of IL-17 were seen in the case group (p = 0.04), whereas there was no significant difference in IL-23 concentrations between the two groups (p = 0.3). The mRNA levels of IL-17 and IL-23 showed no significant differences. In the case group, there was a positive significant correlation between the IL-23 concentration in FF and the oocyte maturation rates (p = 0.01). In the case group, the number of follicles, MII oocytes, immature oocytes, fertilized oocytes and number of embryos were significantly higher than the control group (p < 0.05). Our findings showed that the mRNA expressions of IL-17 and IL-23 were similar in the two groups, and IL-17 was increased in the case group.

Assessing ICSI outcome by combining non-invasive indicators: Early time-lapse morphokinetics and apoptosis in associated cumulus cells among women with the polycystic ovarian syndrome.

Cumulus cells features and embryo developmental events can be considered as noninvasive indicators for embryo selection and clinical outcomes. A combination of time-lapse morphokinetic parameters and cumulus cell apoptosis in women with polycystic ovarian syndrome (PCOS) was evaluated for predicting pregnancy outcome. We assessed a total of 547 embryos from 100 intracytoplasmic sperm injection (ICSI) cycles. Time-lapse records were interpreted in time to pronuclear fading (tPNf), time to 2 to 8 cells (t2-t8), direct cleavage, reverse cleavage, and also for the presence of multinucleation. Percentages of apoptosis were identified in 100 associated cumulus cell samples using the TDT-mediated dUTP-biotin nick end-labeling assay. The significant decrease of apoptotic cumulus cells was detected in patients with chemical and clinical pregnancies as well as live birth among patients PCOS and in the tubal infertility group (p > 0.05). Furthermore, significantly higher implantation rate and also significantly lower cases of early pregnancy loss were observed in the group of oocytes with less apoptotic cumulus cells. Multivariate logistic regression analysis showed that tPNf together with cumulus cell apoptosis were independent prognostic factors of chemical pregnancy, clinical pregnancy rate, and live birth. Time-lapse embryo parameters may not reflect the cumulus cell apoptosis rate. However, the rate of apoptotic cumulus cells is significantly associated with ICSI outcome using Day 3 embryo transfer.