Male partner screening before in vitro fertilization: preselecting patients who require intracytoplasmic sperm injection with the sperm penetration assay.

OBJECTIVES: To determine the diagnostic accuracy of the sperm penetration assay (SPA) and standard semen parameters for subsequent fertilization in in vitro fertilization-embryo transfer (IVF-ET). DESIGN: Prospective study. SETTING: Andrology Laboratory, and university research laboratory. PATIENTS: Two hundred sixteen couples undergoing male-partner screening before IVF-ET (265 cycles). INTERVENTION(S): Male-partner screening (semen analyses [SA] and SPA), standard IVF-ET procedures, follow-up of fertilization in IVF-ET. MAIN OUTCOME MEASURE(S): Diagnostic accuracy of SA and SPA for prediction of fertilization in IVF-ET. RESULT(S): The SPA predicted IVF fertilization with high negative (84%) and positive (98%) predictive rates, and correct prediction in 88% of cycles. In contrast, sperm concentration, motility, morphology, and complete SA showed poor diagnostic accuracy, with correct prediction of IVF fertilization in 64%, 65%, 45%, and 68% of cycles, respectively. CONCLUSION(S): Very low sperm concentration and/or motility were good predictors of poor IVF fertilization, however, low to normal semen parameters were not predictive of successful IVF fertilization. The SPA is a useful screening tool that predicts IVF fertilization with high diagnostic accuracy. The SPA may be useful to discriminate between those couples with a high probability of normal fertilization in IVF and those with a low probability of normal fertilization that may benefit from assisted fertilization by intracytoplasmic sperm injection (ICSI).

Permanent impairment of embryo development by hydrosalpinges.

Recent reports suggest a deleterious effect of hydrosalpinges on pregnancy outcome for in-vitro fertilization (IVF) and improvement following surgical treatment. We compared the effect of hydrosalpinx on pregnancy outcome in 286 patients having 348 IVF cycles and followed the development of untransferred embryos for 7 days to determine if hydrosalpinges affect oocyte quality or embryo development. The delivery rate per retrieval was significantly lower for patients with hydrosalpinx, but was restored by surgical treatment to that of patients without hydrosalpinx. However, the implantation rate per embryo transferred and normal blastulation of untransferred embryos, which were significantly decreased in patients with hydrosalpinx, and growth arrest and degeneration of untransferred embryos, which were significantly increased compared to patients without hydrosalpinx, were not restored by surgical treatment of hydrosalpinges. We conclude that surgical treatment of hydrosalpinges decreases early pregnancy loss and improves pregnancy outcome, possibly by diminishing reversible deleterious effects exerted on the endometrium. As we have seen in our laboratory, hydrosalpinges may have a permanent negative influence on ovarian function, follicular development and oocyte quality since implantation of transferred embryos and normal blastulation of untransferred embryos remain low, and in-vitro growth arrest and degeneration remain high despite surgical treatment of hydrosalpinges.

Granulosa cell co-culture enhances human embryo development and pregnancy rate following in-vitro fertilization.

A preliminary study and related clinical trial were performed to evaluate the effects of granulosa-lutein cell co-culture on human embryo development and pregnancy rates for in-vitro fertilization (IVF). In the study, sibling two-pronuclear zygotes were randomly allocated to culture with (co-culture) or without (control) autologous granulosa-lutein cells. After 24 h, embryos were examined for blastomere number and degree of fragmentation. Co-culture had no effect on the average number of blastomeres per embryo at 24 h; however, fragmentation was significantly decreased in co-cultured embryos (0.7 +/- 0.1) compared with controls (1.3 +/- 0.2; P < 0.05). In the subsequent clinical trial, all two-pronuclear zygotes were co-cultured for 48 h prior to embryo transfer. The live birth rate per embryo transfer was 43.4% with an implantation rate per embryo of 17.6%. Of the untransferred embryos, 68% developed to the blastocyst stage and were cryopreserved. We conclude that the simple system of autologous granulosa-lutein cell co-culture improves embryo development, implantation and subsequent pregnancy rates for IVF.

Growth factor effects on endometrial epithelial cell differentiation and protein synthesis in vitro.

OBJECTIVE: To develop a baseline for projected studies of a rat endometriosis model. DESIGN: We investigated the effects of two macrophage-related growth factors, platelet-derived growth factor (PDGF) and transforming growth factor-beta (TGF-beta), on proliferation, in vitro differentiation, and protein secretion of uterine epithelial cells from immature rats. Uterine epithelial cells grown on matrix covered filters were treated with growth factors (GFs) or estrogen and/or P. Incorporation of [35S]methionine by polarized uterine epithelial cell proteins and secretion of labeled proteins into apical and basal culture medium were examined. SETTING: Department of Cell Biology, Baylor College of Medicine, Houston, Texas. MAIN OUTCOME MEASURES: Cell associated and secreted proteins were resolved by gel electrophoresis, fluorography, and immunoblotting. Proliferation was quantified by cell counts in parallel cultures by hemocytometer. RESULTS: Estrogen and P increase protein synthesis by uterine epithelium. Transforming growth factor-beta depressed protein synthesis and secretion in uterine epithelial cells. Platelet-derived growth factor appears to have no effect on epithelial protein synthesis or secretion and does not modulate the effect of TGF-beta. Estrogen and P increase complement component 3 (C3) production by epithelial cells. CONCLUSION: Macrophage-secreted GFs may play a role in the development and maintenance of ectopic endometrial tissue. Both TGF-beta and ovarian steroids may participate in the dynamic regulation of protein synthesis by ectopic uterine epithelium. These molecules may indirectly affect the macrophage-stromal axis through nonspecific modulation of C3 secretion. Platelet-derived growth factor appears to have no direct effect on uterine epithelial cells. The recognized effect of PDGF on ectopic endometrial tissue is most likely mediated via the stromal component.

Value of diagnostic laparoscopy in young women with possible appendicitis.

Removal of a normal appendix because of suspected appendicitis occurs most frequently in women of reproductive age. We investigated the value of laparoscopy in the diagnostic evaluation for possible appendicitis in women of childbearing age. Fifty-one women were entered in a management protocol, which included diagnostic laparoscopy in instances with atypical features. Twenty patients underwent immediate appendectomy because of history and physical findings classical for appendicitis; 31 women with atypical history and physical findings underwent an initial diagnostic laparoscopy. In the group of patients who underwent immediate appendectomy because of classical presentation, five of the appendices removed were normal. At laparoscopy, appendicitis was diagnosed in five patients, another disease in 15 and no diagnostic abnormality was found in six. Five patients had a normal appendix removed, two because of questionable inflammation and three because of nonvisualization of the appendix. There were no false-negative examination results. Twenty-one patients required no further operative intervention after laparoscopy. The negative appendectomy rate remained unchanged (ten negative appendectomies of 30 performed). Definitive diagnoses of eight instances of pelvic inflammatory disease, six of ruptured ovarian cysts and one instance of ileitis of the small intestine were made earlier than would have been possible without diagnostic laparoscopy in this setting. Diagnostic laparoscopy permits earlier definitive diagnosis and prompt institution of appropriate therapy for disease of the female reproductive tract that simulates appendicitis. Caution is advised, however, when diagnostic laparoscopy is applied more frequently than right lower quadrant exploration in the management of probable appendicitis. The improvement in diagnostic accuracy may be offset by an increased number of negative appendectomies resulting from nonvisualization and false-positive inflammation.