A method of short-term cryostorage and selection of viable sperm for use in the various assisted reproductive techniques.

The objective of this study was to determine if spermatozoa, following short-term cryostorage at 5 degrees C in Test-Yolk buffer (TYB; ZBL, Inc., Lexington, KY, USA), could be recovered and improved via the SpermPrep filtration method and to assess the possibly enhanced fertilizing capacity of the selected spermatozoa. Semen specimens from 20 men were collected, evaluated, diluted 1:1 (v/v) with TYB, divided into aliquots and cooled to 5 degrees C for 24 and 48 hr. Semen samples were assessed for volume, sperm count, percentage and grade of motility, percentage of morphologically normal spermatozoa and outcome of the sperm penetration assay (SPA). After storage, aliquots were rewarmed at 37 degrees C, centrifuged, and the pellet was resuspended in 1.0 ml of SpermPrep media (ZBL, Inc.). Following 15 min of incubation, the rewarmed spermatozoa were filtered via the SpermPrep I filtration column (ZBL, Inc.) and assessed accordingly. The results obtained in this study indicate that the short-term cryostorage procedure yielded spermatozoa of adequate qualitative characteristics when compared to the fresh spermatozoa. Furthermore, filtration of rewarmed specimens yielded spermatozoa of significantly higher qualitative characteristics and superior fertilizing capacity following a short-term cryostorage period in TYB when compared to fresh and rewarmed spermatozoa (p < 0.05). This method of short-term cryostorage in TYB and selection of superior spermatozoa via the SpermPrep filtration method could further enhance the fertilizing ability of patients who produce spermatozoa characterized by deficient capacitation, acrosome reaction and subsequent fertilization.

Differences in seminal parameters in specimens collected via intercourse and incomplete intercourse (coitus interruptus)

Whatever the mechanism of such improvements may be, the results in this study point out that coitus interruptus in the human may not be the method of choice for collection of semen specimens, especially in patients with spermatogenic dysfunctions such as hypospermia, oligospermia, and asthenospermia. It should also be noted at this point that, for whatever purpose (semen analysis or artificial insemination by husband), the collected specimen should as closely as possible resemble the ejaculate delivered during intercourse. The complete coitus method, as applied in this study, showed that completion of the ejaculatory process during intercourse as compared with the coitus interruptus method, may assist in the improvement of the collected specimen and should closely resemble the ejaculate obtained during intercourse without the use of Silastic condoms. Furthermore, on the basis of the results generated in this study, the complete coitus method should always be the method of choice for male infertility patients with ejaculatory and spermatogenic dysfunctions as well as for scientists and clinicians who deal in the field of infertility diagnosis and treatment.

Circadian, ultradian, and episodic gonadotropin and prolactin secretion in human pseudocyesis.

Six women with pseudocyesis were studied by 15-min blood sampling for 12 to 24 h to determine their gonadotropin and PRL secretory profiles aiming to clarify the endocrine alterations in this form of hypothalamic amenorrhea. Clinical and biochemical evidence of hyperandrogenism was found in 4 patients. Persistent hyperprolactinemia was present only in one patient. Significant circadian and ultradian periodicities were identified by time series analysis in the 12-24 h profiles of FSH, LH and PRL secretion. Pulse analysis by the Van Cauter (UL-TRA.JN) method revealed a 24-h mean LH interpulse interval of 91 +/- 21 min with a mean LH amplitude of 5.4 +/- 0.8 IU/l. There was a significantly lower pulse frequency at night than during the daytime. The mean 24-h PRL interpulse interval and pulse amplitude were 134 +/- 22 min and 9.2 +/- 1.8 IU/l, respectively. Both FSH and LH mean levels were higher during the daytime than at night, while the reverse was true for PRL values. Decreased LH pulse frequency and amplitude emerged as the most distinctive findings. Antecedent hypothalamic-pituitary aberrations due to other endocrinopathies and the timing of the hormonal assessment (e.g. recovery phase) may explain, at least in part, the reported heterogeneity of neuroendocrinologic findings in pseudocyesis.

Amniotic fluid beta-endorphin levels and labor.

To define the relationship between amniotic fluid concentrations of beta-endorphin immunoreactivity and onset of parturition, we measured this opioid in samples obtained during cesarean section at term. A total of 27 women were studied, 14 without labor and 13 in early labor. Mean (+/- SE) amniotic fluid beta-endorphin levels were significantly lower in patients in labor than in those not in labor (3.2 +/- 0.05 versus 8.4 +/- 1.0 fmol/mL). The mean beta-endorphin level (21.1 +/- 4.2 fmol/mL) in other amniotic fluid samples obtained during the second trimester of pregnancy was significantly higher than mean values at term. These differences in amniotic fluid beta-endorphin levels may support the theory of an opioid mechanism involved in parturition.

Maternal and fetal beta-endorphin release in response to the stress of labor and delivery.

In order to clarify the stress effect of labor on maternal and neonatal plasma levels of beta-endorphin, we measured this peptide in samples taken from 40 pregnant patients and their neonates at the time of normal vaginal delivery (n = 15), and at cesarean section performed either in early labor (n = 13) or prior to labor (n = 12). The mean (+/- SE) maternal plasma concentration of beta-endorphin in the vaginal delivery group was 40.3 +/- 5.6 fmol/ml, which was significantly higher than that in their neonates (21.3 +/- 2.9 fmol/ml). In contrast, maternal levels of beta-endorphin in the cesarean section groups (8.2 +/- 1.2 and 8.5 +/- fmol/ml) were significantly lower than those in their neonates (23.3 +/- 5.6 and 15.6 +/- 2.8 fmol/ml). Concentrations of beta-endorphin in mothers delivered vaginally were also significantly higher than those in mothers delivered by cesarean section. However, there was no difference in mean cord levels of beta-endorphin among the three groups. These findings indicate that neither the presence or absence of labor affects fetal plasma beta-endorphin secretion and the stress of labor and delivery produces a marked increase in maternal release of beta-endorphin.