The apoptotic pathway in fertile and subfertile men: a case-control and prospective study to examine the impact of merocyanine 540 bodies on ejaculated spermatozoa.

OBJECTIVE: To evaluate the presence of merocyanine 540 (M540) bodies and their impact on the measurement of apoptotic biomarkers in human spermatozoa. DESIGN: Case-control, prospective study. SETTING: Academic centers. PATIENT(S): Fertile and subfertile subjects. INTERVENTION(S): Semen samples from subfertile and fertile men, 11 per group, were analyzed for basic semen parameters and early (annexin-V binding) and late (terminal deoxynucleotidyl transferase dUTP nick end labeling [TUNEL]) sperm apoptotic biomarkers by flow cytometry. Samples were also stained with M540 to assess the presence of M540 apoptotic bodies. MAIN OUTCOME MEASURE(S): Presence of M540 apoptotic bodies. RESULT(S): Groups differed significantly in the expression of early and late apoptosis biomarkers. The percentage of M540 bodies between groups was not different. The exclusion of M540 bodies from TUNEL results did not have a significant impact on measurement in either fertile or subfertile groups. CONCLUSION(S): This study confirmed the occurrence of M540 bodies in semen and that male factor infertility is associated with an increased expression of apoptosis biomarkers. Moreover, we demonstrated that the presence of M540 bodies did not affect the quantification of apoptotic biomarkers in either group.

[Sperm capacitation with double ejaculate of short interval. A choice in assisted reproduction]. / Capacitación espermática con doble eyaculado de corto intervalo. Una alternativa en reproducción asistida.

In the techniques of assisted reproduction a minimum number of spermatozoa are required, but some patients cannot provide them with one capacitated seminal sample. In this study we investigated if in vitro sperm capacitation of two ejaculates obtained one hour apart from patients with reduced sperm amounts provides an adequate number of spermatozoa. Samples of 75 patients with a low seminal account were processed. They provided one second seminal sample after 60 minutes from the first one. The parameters of the capacitated sample of the first ejaculate (PE) were compared with the parameters of the capacitated sample joining the first and the second ejaculates (PSE). A paired t-test was applied considering a significant value of p < 0.05. Concentration, mobility, total of mobile cells (TCM) and total of mobile cells with normal morphology (ICR) were 31.28 +/-17.65 million/mililiter, 50.45 +/-26.3%, 9.06 +/-7.85 million and 0.64 +/- 0.62 million, respectively in first ejaculate. The respective values in first and second ejaculates were 54.45 +/- 35.06 million/mililiter, 59.39 +/- 25.41%, 17.94 +/- 12.18 million and 1.32 +/- 1.11 million, which represents a significant increase (p<0.05) of 74%, 17%, 98% and 106% respectively. Neither the volume (0.49 +/- 0.037 mililiter versus 0.49 +/- 0.019 mililiter, increase 0%) nor morphology (5.66 +/- 2.76% versus 6.11 +/- 4.88%, increase 7.9%) had a significant change. Sperm capacitation with double ejaculate of short interval must be considered for patients with decreased sperm amounts when deciding the reproductive technique.