Increased fertilization rates after in vitro culture of frozen-thawed testicular immotile sperm in nonobstructive azoospermic patients.

Objective. To optimise the use of freeze/thaw testicular immotile spermatozoa from nonobstructive azoospermia patients and to analyse the outcome of intracytoplasmic sperm injection (ICSI) of such spermatozoa. Methods. Testicular specimens were retrieved and cryopreserved from forty patients with nonobstructive azoospermia and underwent one cycle with thawed spermatozoa (Group I) that led to pregnancy in sixteen cases. Twenty-four patients of group I underwent treatment with the same batch of thawed spermatozoa (Group II). For the first ICSI attempt, injection was performed when motile spermatozoa were found. In group II, injection was performed when maximum motility was reached. We compared mean of fertilization rate, embryo quality, clinical pregnancy rate and embryo implantation rate. Results. The mean percentage of motility was significantly higher in the group II than in the group I (18, 6 versus 8, 2). Group I showed a significant decrease in fertilization rates when compared with cryopreserved testicular spermatozoa in group II (54% versus 72%, P < 0.05). No difference was noted between the cleavage rate, embryo quality, clinical pregnancy rates and implantation rates among group II and I. Conclusion. Fecundation rate can be significantly improved after in-vitro culture and sperm selection of frozen-thawed immotile testicular spermatozoa in patients with nonobstructive azoospermia.

Varicocele, leucocitospermia y su impacto en la fragmentación del ADN espermático / Varicocele, leukocytospermia and its impact on the spermatic DNA fragmentation

Introducción: El estrés oxidativo en una de las causas que puede explicar la presencia de unos niveles elevados de daño en la molécula de ADN. En 2 cuadros clínicos que afectan a la línea germinal del varón, la leucocitospermia y el varicocele, se reconoce la incidencia de niveles elevados de estrés oxidativo. Objetivos: En el presente trabajo se compararon 2 cuadros clínicos, varicocele y leucocitospermia, con objeto de comprobar si existe una intensidad diferencial de la fragmentación del ADN espermático. Material y métodos: Se incluyeron como controles externos donantes de espermatozoides con fertilidad probada y pacientes con factor masculino no determinado. A diferencia de otros estudios de fragmentación del ADN espermático, en este caso, se consideraron tanto los niveles de fragmentación absolutos (SDF), como la proporción de espermatozoides degradados en el total de fragmentados (índice de degradación [ID]) de acuerdo con la información obtenida tras la aplicación del test Halosperm. Resultados: Se demostró un aumento muy significativo de la fragmentación del ADN espermático en las muestras seminales de pacientes con varicocele y con leucocitospermia. Los pacientes con varicocele mostraron un ID 2 veces mayor que el observado en pacientes con factor masculino no determinado o en pacientes con leucocitospermia, y 6 veces mayor que en los donantes. Discusión: La presencia de unos niveles de estrés oxidativo elevados podría ser una explicación asumible para justificar los altos niveles de daño observado en los espermatozoides de los pacientes tanto con varicocele como con leucocitospermia, pero probablemente estos niveles sean más elevados en el caso del varicocele ya que los niveles de degradación del ADN espermático son superiores a los observados en cuadros de leucocitospermia... (AU)

Introduction: High levels of oxidative stress can explain the presence of high levels of damage in the DNA molecule. The impact of high levels of oxidative stress in 2 clinical circumstances affecting the male germ line has been well established: leukocytospermia and varicocele. Objective: To assess sperm DNA fragmentation in patients diagnosed with leukocytospermic and varicocele. Material and methods: Leukocytospermic and varicocele patients and external controls (donors with proven fertility and patients with undetermined male factor). Unlike in other studies of sperm DNA fragmentation, in this study both the proportion of damaged sperm after using the sperm chromatin dispersion test (Halosperm), and the proportion of degraded sperm in total fragmented (degradation index [DI]) were taken into consideration. Results: A highly significant increase in sperm DNA fragmentation has been observed in semen samples of patients with varicocele and leukocytospermia. Varicocele patients showed a DI twice as high as that observed in patients with undetermined male factor or in patients with leukocytospermia, and 6 times as high as that observed in the donors. Discussion: The presence of high levels of oxidative stress could be an acceptable explanation for the high levels of damage observed in the spermatozoa of varicocele patients or with leukocytospermia; the level of sperm degradation is higher in the case of varicocele than those observed in leukocytospermia. Conclusions: SDF levels in patients with leukocytospermia and varicocele are significantly higher than those observed in donors or men with undetermined male factor. The DI in sperm samples from patients with varicocele is the highest of all the samples studied in this analysis. The routine determination of the DI may have a practical value, by guiding the patient towards the potential diagnosis of varicocele, even when this is subclinical (AU)

Ureaplasma urealyticum reduces motility and induces membrane alterations in human spermatozoa.

The in-vitro effects of several concentrations of Ureaplasma urealyticum on the motility, membrane integrity and morphology of washed spermatozoa from healthy donors were studied. A significant reduction in sperm motility and signs of membrane alteration, directly related to U.urealyticum concentration and contact time were observed. Scanning electron microscopy examination showed masses of U. urealyticum attached to the head and middle piece of some of deformed spermatozoa. It is suggested that U.urealyticum is involved in sperm changes leading to male infertility, particularly when there is heavy U. urealyticum colonization or specific infections with this microorganism.