Relationship between male reproductive hormones, sperm DNA damage and markers of oxidative stress in infertility.

This study investigated the relationship between male reproductive hormones and sperm DNA damage and markers of oxidative stress in men undergoing infertility evaluation for male factor (n = 66) and non-male factor (n = 63) infertility. Semen samples were analysed for DNA fragmentation index (DFI). Serum samples were analysed for FSH, inhibin B, anti-Müllerian hormone (AMH), testosterone and total antioxidant capacity (TAC). Serum inhibin B was significantly lower in the male factor group compared with the non-male factor group. Inhibin B showed a positive correlation with sperm concentration and motility, and serum AMH showed a positive correlation with sperm concentration and semen volume. DFI was 3-fold higher in the male factor group and showed a negative correlation with sperm motility. Blood plasma TAC was negatively related to sperm concentration. The results confirm that AMH and inhibin B are markers of Sertoli cell function. Sperm DNA damage is moderately increased in male factor infertility, and is negatively associated with sperm motility. A negative association between antioxidant activity and sperm concentration suggests that even minimal oxidative stress may influence sperm concentration. However, there was no significant relationship between hormone concentrations, sperm DNA damage and total antioxidant capacity, suggesting other mechanisms for sperm dysfunction.

Antenatal ultrasound and magnetic resonance imaging in localizing the level of lesion in spina bifida and correlation with postnatal outcome.

OBJECTIVE: To compare prenatal ultrasonography and magnetic resonance imaging (MRI) in the diagnosis of spina bifida with postnatal MRI/surgical findings and to study the postnatal outcome in relation to the level of lesion and head signs on antenatal imaging. METHODS: A retrospective study of babies referred to a tertiary neurosurgical unit with a diagnosis of spina bifida in the years 2000-2002 was performed. The levels of lesions and head signs diagnosed using antenatal ultrasonography and MRI were compared with postnatal MRI and operative findings. The levels of lesions and head signs diagnosed pre- and postnatally were used to study the correlation with neurological outcome at a mean follow-up period of 12 months. RESULTS: Twelve antenatally diagnosed and five postnatally diagnosed cases of spina bifida were seen. The level of lesion as identified by antenatal ultrasound correlated with that observed postnatally in 8/12 cases and the antenatal ultrasound diagnosis of ventriculomegaly was confirmed postnatally in all cases. The level of lesion as identified by antenatal MRI correlated with that observed postnatally in 4/8 cases (50%) and the antenatal MRI diagnosis of ventriculomegaly was confirmed postnatally in 5/8 (63%) cases. 12/17 babies were found to have residuals in the immediate postnatal urodynamic studies, of which 83% (n = 10) required intermittent catheterization of the bladder at 12 months of age. Low spinal lesions were associated with increased bladder morbidity when compared to high spinal lesions (8/10 vs. 4/7, P < 0.05) (odds ratio (OR) = 10.0; 95% CI, 1.05-95.01). The level of spinal lesion and the presence of ventriculomegaly did not have any statistically significant correlation with motor functions, morbidity and developmental milestones. CONCLUSIONS: Antenatal ultrasonography is a good tool in the diagnosis of spina bifida. We could not demonstrate any advantage of antenatal MRI over ultrasonography. There is better correlation of ultrasonography than MRI with postnatal MRI/operative findings in terms of level of lesion and head signs. Low spinal lesions appear to be associated with increased bladder morbidity.

Development of a sensitive enzyme immunoassay for anti-Müllerian hormone and the evaluation of potential clinical applications in males and females.

BACKGROUND AND OBJECTIVE: Recent studies have found anti-Müllerian hormone (AMH) to be a potentially important marker for the assessment of ovarian reserve and prediction of the success of in vitro fertilization (IVF) treatment. The objectives of this study were to develop a sensitive and specific assay for AMH and to evaluate the potential application of the assay. This assay will be then available to our collaborators in the UK and overseas. DESIGN: Samples obtained as part of another prospective cross-sectional study from infertility patients and another prospective longitudinal study from pregnant women were used in this study to measure AMH using a new double-antibody enzyme-linked immunosorbent assay (ELISA). PATIENTS AND MEASUREMENTS: AMH levels were evaluated in (i) serum and seminal fluid from males (normal and male factor infertility males), (ii) serum and follicular fluid from females (normal and female with unexplained infertility) and (iii) serum, amniotic fluid (AF) and coelomic fluid (CF) from pregnant women. AMH levels in the samples were measured by a newly developed ELISA. RESULT: The assay had a detection limit of<0.078 ng/ml. High recoveries of spiked recombinant protein were observed from male and female sera and also from follicular, seminal, coelomic and amniotic fluids. The intra- and interassay coefficients of variation (CVs) were 3.6% and 4.0%, respectively. Serially diluted human samples gave dose-response curves parallel to the standard curve. Immunoreactivity was stable to sample storage at room temperature for several days and to multiple cycles of freezing and thawing. In seminal fluid, the AMH concentrations in a group of men with male factor infertility were insignificantly different from those in fertile men. By contrast, serum AMH concentrations were lower in the male factor infertility group than the normal group of patients. Women with unexplained infertility had similar concentrations of AMH in serum and follicular fluid compared to controls. Pregnant women had higher concentrations of AMH in the circulation in early pregnancy compared with nonpregnant women, suggesting a foeto-placental contribution and a possible biological role for this molecule in early pregnancy. CONCLUSION: We have developed a sensitive and specific assay for AMH. Serum AMH in men with male factor infertility is lower than in normal men. Levels of AMH in pregnancy are higher than normal menstrual cycle levels suggesting a foeto-placental contribution.