COMET, TUNEL, and TEM analysis of an infertile male with short tail sperm.

Male infertility is correlated with sperm morphology and sperm DNA damage, which are completely different from that of fertile individuals. An accurate sperm DNA damage analysis and ultrastructural examination of the ejaculate provide important support in the clinical evaluation. It is supposed that in the near future, the fertilization rate, pregnancy rate, and miscarriages could be predicted using the combination of these types of tests in assisted reproductive technologies (ARTs). For this purpose, we report a very rare case of an infertile man having short tail sperm. The infertile man and his wife underwent in vitro fertilization (IVF) with intracytoplasmic sperm injection (ICSI). During this process, we examined the ultrastructure of the ejaculated sperm with transmission electron microscopy (TEM) and calculated the sperm DNA damage with terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) and COMET assays. Then, we evaluated the association between sperm DNA damage and embryo quality.

Fluctuation of "sperm DNA integrity" in accordance with semen parameters, and it's relationship with infertility.

PURPOSE: Nowadays sperm analysis is used routinely to assess infertile men, but it's predictive value is still restricted. For this reason, new markers are needed for diagnosis and guide treatment. A DNA damage evaluation method, which is easily applicable for routine analysis is described in this paper to provide an important support in clinical evaluation. METHODS: In this study, DNA damage evaluation was planned in infertile males with the COMET assay, a method which is simple, cheap, sensitive and reliable. For this purpose, the sperm DNA damage of normospermic, oligospermic, asthenospermic and teratospermic cases were investigated and the relationship between sperm parameters (sperm count, motility, morphology) and DNA damage was assessed. RESULTS AND CONCLUSION: According to our results we suggest that the factors to blame for sperm DNA damage and semen quality are the same. Also it was found that the infertile males possess substantially more sperm DNA damage than fertile men do. Improving and using the DNA damage evaluation tests seem to improve the results in ART clinics.

Homocysteine effects on cellular glutathione peroxidase (GPx-1) activity under in vitro conditions.

Hyperhomocysteinemia is associated with a lot of diseases including cardiovascular diseases and neural tube defect, but it has not been clarified exactly which mechanism is responsible for occurence disease. Here, homocysteine (Hcy) and cysteine (Cys), which are thiol containing amino acids, were examined for their effect on glutathione peroxidase (GPx) activity. It was observed that the GPx-1 activity was inhibited under severe hyperhomocysteinemia (50-500 microM Hcy) conditions, especially at low glutathione concentrations but that cysteine increased GPx-1 activity at low glutathione concentrations and inhibition clearly appeared at 500 microM Cys concentration.