Retrograde Horseradish Peroxidase Transport in Motor Axons after Nd:YAG Laser Irradiation of the Tibial Nerve in Rats

We have recently demonstrated that the number of small sensory neurons of the A-δ- and C-fiber group in lumbar dorsal root ganglia labeled with horseradish peroxidase (HRP) is selectively decreased 7 days after Nd:YAG laser irradiation of the tibial nerve in the rat. In contrast, the number of large diameter sensory neurons was not affected by laser application. In an attempt to clarify the fate of motoneurons after laser irradiation of their peripheral axons, the numbers of lumbar motoneurons retrogradely labeled with HRP 7 days after Nd:YAG laser irradiation of the tibial nerve have been determined in rats. Our results show that the number of HRP-labeled motoneurons in lumbar segments L6 to L3 is not altered to a significant extent after laser irradiation of their peripheral axons (laser-treated side, 767 ± 10 cells vs control side, 808 ± 19; n = 5, mean ± SEM). In addition, no difference was detected in the mean value or the distribution of soma cross-sectional areas of labeled motoneurons on the laser-treated side and the control side. Specifically, the numbers of HRP-labeled small diameter motoneurons, which are presumably γ in type and have a conduction velocity similar to sensory neurons of the A-δ group, were not affected by laser application. Possible mechanisms of the differential vulnerability of sensory neurons as compared to motoneurons of similar size are discussed.     

Preliminary findings in germinal vesicle transplantation of immature human oocytes

Transplanting a germinal vesicle (GV) from an aged woman's oocyte into a younger ooplasm has been proposed as a possible way to reduce the incidence of oocyte aneuploidy which is considered to be responsible for age-related infertility. In this study, we have assessed the efficiency of each step involved in nuclear transplantation-specifically cell survival, nuclear-cytoplasmic reconstitution, and the capacity of the reconstituted oocytes for in-vitro maturation. In addition, we have evaluated the fertilizability and karyotypic status of the manipulated oocytes by intracytoplasmic sperm injection (ICSI) and fluorescent in-situ hybridization technique respectively. Nuclear transplantation was accomplished with an overall efficiency of 73%. Due to the limited availability of materials, most nuclear transplantation procedures were performed between sibling oocytes. The maturation rate of 62% following reconstitution was comparable with that of control oocytes, as was the incidence of aneuploidy among the reconstituted oocytes. The ICSI results of the reconstituted oocytes yielded a survival rate of 77%, a fertilization rate of 52%, and a satisfactory early embryonic cleavage. Furthermore, in a limited number of observations where the nucleus of an aged oocyte was transferred into a younger ooplasm, there was an appropriate chromosomal segregation. These findings demonstrate that human oocytes reconstituted with GV nuclei are able to undergo maturation, fertilization, and early embryo cleavage, and maintain a normal ploidy. Although in-vitro maturation seems to be a limiting step, this technique would allow us to investigate further the nuclear-ooplasmic relationship during meiotic maturation.     

Late luteal estradiol patterns are a better prognosticator of pregnancy outcome than serial beta-human chorionic gonadotropin concentrations

OBJECTIVE: Since the corpus luteum (CL) is known to play an important role in early pregnancy, its activity could possibly be a marker for pregnancy outcome. DESIGN: The late estradiol (E2) concentration in 48 viable pregnancies and 39 pregnancies which resulted in spontaneous abortions after in vitro fertilization and embryo transfer were used to evaluate such predictability. SETTING: All patients studied were of the Center for Reproductive Medicine at Cornell University Medical College. PATIENTS, PARTICIPANTS: Eighty-seven patients. INTERVENTIONS: None. MAIN OUTCOME MEASURE: Serum E2 and human chorionic gonadotropin (hCG) concentrations on day +11, +13, +15 (day +1 = day of ovum pick-up) were measured and studied. RESULTS: The late luteal CL activity after rescue had a positive correlation with the number and quality of the implanted embryos. Reduced CL activity was indicative of abortion. The late luteal E2 pattern when compared with hCG doubling time had a better abortion predictability (37.8% versus 63.9%, respectively). CONCLUSION: Corpus luteum activity demonstrated to be a better prognosticator of abortion than serial beta-hCG titers.     

Pregnancy related to infertility diagnosis, number of attempts, and age in a program of in vitro fertilization

Three hundred nineteen infertility patients who did not become pregnant with standard therapies presented to Norfolk for in vitro fertilization between January 1981 and December 1983. There were 560 laparoscopic cycles with 105 pregnancies. The pregnancy rate was 18.8% by cycle, 24.5% by transfer, and 33% by patient. The overall pregnancy rate was found to be independent of infertility diagnosis, age, and number of attempts.     

The perimenopausal patient in in vitro fertilization: the use of gonadotropin-releasing hormone

The perimenopause, incipient ovarian failure, is a major problem in stimulation failures during an in vitro fertilization program. This must be recognized as not necessarily related to age but also associated with adnexal inflammatory and operative processes. Although ovulation occurs uninterruptedly, the follicle-stimulating hormone in the early follicular phase is elevated and the luteinizing hormone is normal. Characteristically, there is no estradiol response to human menopausal gonadotropin therapy or a rapid response with a premature luteinizing hormone surge. These problems sometimes may be overcome with pulsatile intravenous gonadotropin-releasing hormone therapy, 5 or 10 micrograms/90 or 120 minutes. The major therapeutic problem is in the identification of a luteinizing hormone surge in these patients. Of eight women who were treated, two failed to respond with follicular maturation, three either had no oocytes aspirated from apparently postmature follicles or had postmature oocytes; and one had treatment cancelled due to ovulation. The four latter patients may have failed because of unrecognized ovulation. In the remaining two patients, one oocyte was fertilized and transferred, and one pregnancy occurred.     

Transdermal estrogen replacement in ovarian failure for ovum donation

This study examined the efficacy of transdermal estradiol (TE2) replacement versus oral estradiol (OE2) through evaluation of peripheral steroid levels, endometrial morphology, and clinical outcome in six patients with ovarian failure. Patients were begun on sequential E2 and progesterone replacement with transdermal E2 patches. Endometrial biopsies were done on day 21 of the first replacement cycle and day 26 of the second cycle. Controls were 28 cycles on a regular 28-day micronized OE2 protocol. No significant difference was found between E2 levels throughout the cycle of the two respective stimulation protocols, except for days 12 to 14, when the OE2 protocol produced significantly lower E2 than did the TE2 protocol (P less than 0.01). A positive, highly significant correlation was found between estrone (E1) and E2 values in the OE2 group (r = 0.92) (P less than 0.003). During OE2 administration, E1 was significantly higher than E2 (P less than 0.01). E1 was not found to be higher than E2 in the TE2 group, resulting in a significant difference in the E2/E1 ratio of 1.59 +/- 1.6 for TE2 compared with 0.13 +/- .04 for OE2 (P less than 0.05). Early biopsies in patients on TE2 revealed glandular components that were dated as day 18.2 +/- 1.7, while the stroma was dated as day 21.8 +/- 0.8, a statistically significant disparity (P less than 0.01). In patients on OE2, the same significant 3-day glandular/stromal disparity was observed (P less than 0.05). Morphologic evaluation of late biopsy specimens revealed day 25.0 +/- 0.8 and 24.5 +/- 1.5 for TE2 and OE2 groups, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)     

In vitro fertilization and embryo transfer (IVF/ET): An established and successful therapy for endometriosis

The purpose of this report is to present a 6-year experience in the management of endometriosis with in vitro fertilization and embryo transfer (IVF/ET). We divided 136 patients who underwent 280 cycles into three groups: (1) previous history of endometriosis but normal pelvis at the time of oocyte retrieval, (2) stages I–II endometriosis (revised AFS classification), and (3) stages III–IV endometriosis. The stimulation protocols, estradiol (E₂) responses, and distribution of terminal E₂ patterns were similar in all groups. Group 3 had significantly fewer preovulatory and immature oocytes retrieved and fewer embryos transferred. The fertilization rate and the per cycle/per transfer pregnancy rates were similar in all groups. The miscarriage rate was higher in group 3, and the on-going pregnancy rate per cycle was lower, Luteal phase E₂ and progesterone levels were comparable in all groups. No differences were found when groups 2 and 3 were analyzed for the presence of one or two ovaries or the presencelabsence of ovarian endometriosis. The overall fertilization rate, the per cycle/per transfer pregnancy rates, and the miscarriage rate were similar to those of tubal factor patients. We underscore the excellent out-come of patients with minimal or mild endometriosis in IVF/ET. We conclude that patients with moderate or severe endometriosis have a compromised reproductive potential, probably because of a reduced oocyte recovery rate and poor embryo quality.     

Endometrial secretory proteins enhance early embryo development

Purpose To evaluate the role of endometrial stromal cells and their secretory proteins in early embryo development, two-celled CB6F1 mouse embryos were cultured alone or cocultured with human endometrial stromal cells in various culture conditions.Results The percentage of embryo blastocyst formation, hatching, and outgrowth was significantly greater in (1) coculture with endometrial stromal cells than in a cell-free control when both coculture and control were carried out in protein-free medium or in RPMI 1640 plus 10% fetal calf serum; (2) coculture with hormone (i.e., progesterone plus relaxin)-treated cells than in coculture with hormone-nontreated cells; and (3) media supplemented with isolated endometrial secretory proteins than in media supplemented with BSA (0.35%). Embryo development was not found to be significantly different in coculture and in media supplemented with endometrial secretory protein.Conclusion Our data provides credence to the theory that endometrial stromal cells enhance embryo development by secreting specific proteins that are beneficial to embryo growth in vitro.Presented at the 48th Annual Meeting of the American Fertility Society, New Orleans, Louisiana, October 31–November 5, 1992.