Gerald M. Scholl

Monozygotic twinning associated with mechanical assisted hatching

OBJECTIVE To determine the effect of mechanical assisted hatching on the pregnancy rate (PR). DESIGN A retrospective comparative analysis of hatched versus nonhatched consecutive assisted reproductive technology (ART) cycles. SETTING A hospital-based ART program. PATIENT(S) Patients undergoing ART treatment with assisted hatching (1994-1996) were compared with patients who did not have assisted hatching (1990-1993). INTERVENTION(S) None. MAIN OUTCOME MEASURE(S) Pregnancy rate, multiple PR, and rate of monozygotic twinning. RESULT(S) With hatching, the clinical PR per ET increased from 25.2% to 37.1% and the multiple PR per ET increased from 6.8% to 13.1%. In the nonhatched series, there were no monozygotic twins compared with eight cases in the hatched series (1.2% per ET). CONCLUSION(S) Mechanical assisted hatching increases the PR but concomitantly elevates the rate of multiple gestation and multiple gestation of high order. There is a particularly high risk of monozygotic twinning with mechanical hatching.

Intrauterine insemination as treatment for antisperm antibodies in the female

Ninty-one women with long-standing infertility in the presence of humoral antisperm antibodies (ASA) underwent 473cycles of intrauterine insemination of washed sperm (IUI), resulting in 26 pregnancies. Nine pregnancies were achieved in 67 women who underwent 285 IUI during unstimulated cycles (13% pregnancy rate; 3% pregnancy/cycles of treatment). Twenty women underwent 86 IUI after clomiphene citrate (CC) stimulation, resulting in 6 pregnancies (30%; 7% per cycle), while 28 underwent 102cycles of IUI after human menopausal gonadotropins (hMG) stimulation, resulting in 11 pregnancies (39%; 11% per cycle). Twenty-one of the 26 pregnancies (81%) were achieved in the first 2 IUI cycles. During unstimulated and CC IUI cycles, all pregnancies occurred in the first two cycles of treatment, while with hMG stimulation pregnancies also occurred in the third, fourth, and fifth IUI cycles. Nine of these 91 women subsequently conceived spontaneously, and three others conceived through in vitro fertilization. Only 6 of the 38 pregnancies resulted in spontaneous abortion (16%). Thus, pregnancies achieved in women with ASA have no increased risk of abortion.

Mannose ligand receptor assay as a test to predict fertilization in vitro: a prospective study

OBJECTIVE To assess whether mannose receptor assays can predict fertilization outcome in vitro. DESIGN A prospective, double-blind study of the mannose receptor properties of spermatozoa. SETTING Assisted human reproduction program at a university hospital. PATIENT(S) Partners of 140 consecutive women undergoing their first in vitro fertilization cycle. INTERVENTION(S) Motile sperm populations were tested for surface receptors for mannose by measuring their ability to bind fluorescein-labeled mannosylated albumin and to undergo a free mannose-induced acrosome reaction as judged by Pisum, sativum agglutinin binding. MAIN OUTCOME MEASURE(S) Mannose receptor assay results were correlated with fertilization outcomes using several statistical tests, including the chi2 test, chi2 for proportions, t-tests, analysis of variance with Student-Newman-Keuls tests and correlational and receiver operating characteristic (ROC) curve analysis. RESULT(S) The fractional increment increase on incubation in the percent of sperm binding mannose ligand over an intact acrosome correlated with fertilization rates in vitro. Threshold values of mannose ligand binding and of mannose-induced acrosome reactions predictive of fertilization rates were identified by ROC curve analysis. Men were thus classified into one of four groups with differing fertilization rates in vitro. CONCLUSION(S) The increment increase in sperm surface mannose ligand binding by acrosome-intact sperm correctly predicts high and low fertilization rates in vitro and identifies cases where conventional insemination can result in failed fertilization.

A retroprospective study comparing three different assisted hatching techniques

The purpose of this study was to determine the effect of four different assisted hatching techniques on pregnancy rates in women with prior IVF failure in fresh IVF cycles. The results suggested that assisted hatching utilizing laser, chemical, or microsurgical techniques increases both implantation and pregnancy rates.

The effect of calcium ion channel blockers on sperm fertilization potential*†*Supported in part by an office based research grant from the American Foundation for Urologic Disease with funds contributed by Searle.†The Society for Assisted Reproductive Technology Prize Paper presented at the 49th Annual Meeting of The American Fertility Society, Montreal, Quebec, Canada, October 11 to 14, 1993.

OBJECTIVE To evaluate the effects of calcium ion (Ca2+) channel blockers on male fertility potential. DESIGN A case comparison of the surface expression of mannose-ligand receptors on motile spermatozoa from 10 known fertile males and from 10 normospermic men taking Ca2+ channel blockers who were seeking infertility treatment. Examination of the effects of in vitro exposure of sperm from fertile donors (n = 14) to antihypertensive medications. SETTING Patients from a successful university hospital-based IVF-assisted reproductive technology program and from a male urology private practice. INTERVENTIONS Prescription of alternate hypotensive medications for four male patients; cholesterol loading and unloading in vitro of fertile donor sperm. MAIN OUTCOME MEASURES Motile sperm were tested for their ability to bind fluorescein isothiocyanate-labeled, mannosylated bovine serum albumin as an index of the surface expression of mannose-ligand receptors associated with fertility potential. Acrosome status was simultaneously evaluated by fluorescence microscopy with rhodamine-labeled Pisum sativum lectin. Sperm were assayed before and after an 18-hour or 3-day incubation under capacitating conditions in vitro. RESULTS Motile spermatozoa of normospermic men taking calcium antagonists for hypertension control do not express head-directed mannose-ligand receptors at high frequency, nor do they undergo spontaneous acrosome loss. Unexpectedly, mannose-ligand receptor translocation from the subplasmalemmal space over the acrosome to the sperm surface and aggregation over the equatorial-postacrosomal regions occurred in acrosome-intact sperm. This differs from fertile controls in whom receptor translocation to the equatorial-postacrosomal segment is coupled with the acrosome reaction (AR). Discontinuation of calcium antagonists results in complete recovery of parameters associated with sperm fertilizing potential: time-dependent increases in the percentages of spermatozoa exhibiting surface mannose-ligand binding and spontaneous ARs in vitro. The effects of in vivo administration of calcium antagonists is mimicked in control fertile donor sperm by inclusion of a Ca2+ channel blocker in the media employed during capacitating incubations. CONCLUSIONS Therapeutic administrations of calcium antagonists for hypertension control cause reversible male infertility associated with an IVF failure. A mechanism of inhibition of sperm fertilizing potential through insertion of lipophilic calcium ion antagonists into the lipid bilayer of the sperm plasma membrane is consistent with our in vitro studies.

Human sperm fertilizing potential in vitro is correlated with differential expression of a head-specific mannose-ligand receptor**Supported in part by grant RR05924 from the National Center for Research Resources Biomedical Research Support Program, National Institutes of Health, Bethesda, Maryland.††Presented in part at the 39th Annual Meeting of the Society for Gynecologic Investigation, San Antonio, Texas, March 18 to 21, 1992.

OBJECTIVE To determine whether different patterns of surface expression of mannose-ligand binding sites are correlated with capacitation and predictive of the ability of human sperm to recognize and fertilize eggs in vitro. DESIGN Analysis of motile sperm populations (from fertile donors, males presenting for routine semen analysis, and men undergoing IVF) before and after incubation in capacitating media. SETTING Patients from an infertility practice at a major university hospital. INTERVENTIONS None. MAIN OUTCOME MEASURES The mannose ligand-binding capacity of sperm populations were initially assayed by solid-phase mannosylated polyacrylamide bead binding studies. Sperm surface D-mannose binding sites thus detected were localized and visualized by fluorescence microscopy after reaction with a mannosylated fluorescein isothiocyanate-labeled neoglycoprotein probe. Results were correlated with acrosomal status, reproductive histories, and IVF outcomes. RESULTS The percent of sperm with head-directed surface expression of a mannose-specific receptor was increased in fertile donors and males exhibiting normal fertilization in IVF after incubation in albumin-supplemented Hams F-10 medium (GIBCO Laboratories, Grand Island, NY). In normospermic males exhibiting zona binding failure in IVF, mannose-specific receptor was observed over the head surface of few incubated sperm. CONCLUSIONS The appearance of D-mannose-ligand binding sites on the surface of heads of human spermatozoa is associated with zona binding ability in IVF and is a putative determinant in human gamete recognition and fertilization.

Capacitated sperm cells react with different types of antisperm antibodies than fresh ejaculated sperm**Presented at the 45th Annual Meeting of The American Fertility Society, San Francisco, California, November 13 to 16, 1989.

OBJECTIVE To determine if sera of some women have antibodies against capacitated but not freshly ejaculated sperm. DESIGN The sera of 66 women undergoing in vitro fertilization (IVF) were tested for sperm antibodies after 1 hour and 18 hours of sperm incubation in the maternal sera. Subsequently, 5 sera were tested with capacitated versus noncapacitated sperm cells. SETTING The study was carried out in a university hospital department. PATIENTS, PARTICIPANTS The patients were 66 consecutive couples undergoing IVF. INTERVENTIONS Sera and semen that were taken for routine tests as part of the IVF procedures were used. MAIN OUTCOME MEASURES A case with IVF failure associated with late appearance of sperm antibodies prompted us to study the detection of sperm antibodies after 1 hour and 18 hours incubation. RESULTS Of 37 cases negative for sperm antibodies after 1 hour incubation, 7 demonstrated high levels of antibodies after 18 hours incubation. In 21 of 23 cases with low or intermediate levels of antibodies after 1 hour incubation, significantly higher levels (P less than 0.05) of antibodies were found after 18 hours. Different and higher levels of sperm antibodies were observed in five sera after incubation of 1 hour with capacitated sperm as compared with noncapacitated controls. CONCLUSIONS Major antigenic differences may exist between capacitated and noncapacitated sperm. In some women sperm antibodies are reactive against capacitated sperm only. This has no certain clinical significance but may explain certain cases of IVF failure, unexplained infertility, and part of the variation in sperm antibodies testing methods.

In vitro fertilization and embryo transfer at North Shore University Hospital: Initial experiences

SummaryNo technical difficulties were encountered in visualizing follicles and obtaining aspirates at laparoscopy. At least one oocyte was aspirated in 90% of these women. While the majority of eggs inseminated subsequently cleaved (73%), no pregnancies occurred despite seemingly atraumatic embryo transfer in nearly all women. As two pronuclei were visualized in the majority of cases, one cannot invoke polyspermy as a cause of failure of subsequent embryo development; nor is it possible at this time to segregate those abnormalities in the egg, at the time of oocyte retrieval, from those encountered by normal preimplantation embryos in an abnormal uterine environment, perhaps secondary to hormonal aberrations coincident with gonadotropin stimulus.