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[Inhibition of ova fertilizability by pseudolaric acid B in hamster].

Pseudolaric acid B (PA), a diterpenoid compound isolated from the root of Pseudolarix kaempferi Gorden, was injected into hamster ovarian bursa with various concentration before ovulation. The successful rate of fertilization of ova was significantly decreased, but no effect was observed on spermatozoa activity and fertilizing ability. Hamster ova with or without cumulus were treated with PA at a concentration higher than 50 micrograms/ml in the medium, the fertilizing rate of ova was reduced markedly. At the concentration of 5 micrograms/ml, only the capacity of fertilization of the cumulus-free ova was inhibited. When PA was injected ig 20 mg/kg daily to hamsters (female) for 4 d before mating, partial antifertility effect was observed.

Abortifacient Agents↗

Evaluation of human sperm morphology using strict criteria after Diff-Quik staining: correlation of morphology with fertilization in vitro.

New, very strict criteria were used after Diff-Quik staining for evaluating sperm morphology. The results of morphology scoring were correlated with the fertilization rate in vitro. Semen samples from 64 men participating in an in-vitro fertilization programme were used for this study. All men had to have a sperm concentration of greater than or equal to 20 million/ml and a progressive motility of greater than 30%. The morphology evaluation using strict criteria was performed on the same aliquot of semen as that used for in-vitro insemination. If strict criteria showed that normal morphology was less than or equal to 4%, the fertilization rate per oocyte was 23%. If normal morphology was greater than or equal to 11%, 77% fertilization occurred. For proportions of normal morphology between 4 and 11%, the fertilization rate per oocyte was 59% (P less than 0.000001). Among all these morphology groups, classical semen parameters, such as the mean volume, the mean concentration and the mean motility, did not differ significantly, except for the morphology evaluation using WHO criteria. The correlation with fertilization was better for morphology evaluation using strict criteria than for WHO. In conclusion, the method of evaluating sperm morphology based on very strict criteria allows a more accurate prediction of the chance of fertilization in vitro. Further studies should be done to establish the most appropriate cut-off points for severely impaired, intermediate and high fertilization rates.

Azure Stains↗

In vitro sperm penetration speed and its relationship with in vivo bull fertility.

The in vitro tests utilized to evaluate sperm quality represent an interesting and important approach to evaluate ejaculated fecundant capacity. In the present work, the oocyte-penetrating ability of sperm from two bulls with low and two with high in vivo fertility rate was investigated. Sperm-quality parameters, such as sperm concentration, total and progressive motility, acrosome and total sperm anomalies, proximal cytoplasmic drops and live : dead sperm ratio were assessed and batches of sperm homologues in these parameters were selected. In expt 1, a sperm : oocyte ratio of 3000 was used and the oocytes were fixed 5, 10, 15 and 18 h post-insemination (hpi). In expt 2, the sperm : oocyte ratio was reduced to 1000 and the eggs were fixed at 8 and 10 hpi. The results, analysed by chi-square test, showed significant differences (p <0.001) among bulls at 15 hpi in expt 1 and 8 hpi in expt 2; nevertheless, no accordance was found between sperm penetration rate and in vivo fertility. At 18 hpi, the low-fertility bulls exceeded the two high-fertility bulls, supporting previous reports that suggest an opposite correlation between in vivo and in vitro fertility rate at low doses of heparin. Furthermore, a more efficient zona binding ability of one high-fertility bull was pointed out in expt 2 after the reduction of sperm : oocyte ratio, as it reached the highest percentage of penetration when compared with all the others.

Acrosome↗

Countercyclical U.S. fertility and its implications.

The following article is the verbatim text of a report based on research funded by the Social Security Administration and the National Institute of Child Health and Human Development to the Rand Corporation. The report looks at changing fertility rates in the United States and their implications for future population size and age distributions. An economic model of fertility rate is used to explain observed differences in fertility rates amond couples and to predict future rates. The focus is on trends since 1947 because post-World War II data are the most complete. Several explanations for changing fertility rates are examined, and their usefulness in predicting the future is evaluated.

Attitude↗

Outcome of in vitro fertilization/intracytoplasmic sperm injection after laparoscopic cystectomy for endometriomas.

OBJECTIVE: To assess the impact of prior unilateral or bilateral endometrioma cystectomy on controlled ovarian hyperstimulation (COH) and intracytoplasmic sperm injection (ICSI) outcome. DESIGN: Retrospective case-control study. SETTING: Department of Obstetrics and Gynecology, School of Medicine, Hacettepe University, Ankara, Turkey. PATIENT(S): Fifty-seven consecutive infertile patients were enrolled who had previously undergone unilateral (n = 34) or bilateral (n = 23) laparoscopic cystectomy for endometriomas more than 3 cm in diameter and underwent ICSI. The control group consisted of 99 patients with tubal factor infertility. INTERVENTION(S): Controlled ovarian hyperstimulation and ICSI. MAIN OUTCOME MEASURE(S): Cycle cancellation rate, number of oocytes, fertilization rate, embryo quality, clinical pregnancy rate (PR), and implantation rate. RESULT(S): The mean number of oocytes, metaphase II oocytes, and two-pronucleated oocytes were significantly lower in the bilateral cystectomy group compared to the unilateral cystectomy and control groups. However, all other parameters, including fertilization rate, the mean number of embryos transferred, the mean number of grade 1 embryos transferred, the clinical PR per embryo transfer, and implantation rate, were comparable among the three groups. Within the unilateral cystectomy group, the mean number of oocyte retrieved from the operated site was significantly less than in the contralateral nonoperated site. CONCLUSION(S): Laparoscopic endometrioma cystectomy does reduce the ovarian reserve. However, diminished ovarian reserve does not translate into impaired pregnancy outcome.

Adult↗

Caffeine in fertilization medium is not essential for bovine IVF by fully capacitated spermatozoa.

The purpose of this study was to examine: 1) whether caffeine in the fertilization medium under mineral oil is essential for bovine in vitro fertilization by fully capacitated spermatozoa, 2) the minimum concentration of caffeine that shows an adverse effect on the motility of preincubated spermatozoa. Cumulus-oocyte complexes with heterogeneous-appearing ooplasm were matured in in vitro culture for 24 h and used for insemination. The fertilization rates of the preincubated spermatozoa introduced into the fertilization medium containing 0 mM or 5 mM caffeine were examined. The fertilization rate of the spermatozoa introduced into the medium without caffeine (final concentration of caffeine at fertilization was 0.27-0.35 mM) was significantly higher than that in the medium with 5 mM caffeine (82.4% vs 55.2%, P<0.05). When the final concentration of caffeine at fertilization was reduced ten-fold (0.02-0.03 mM), the fertilization rate was not significantly improved (86.0%). The motility of the preincubated spermatozoa introduced into the fertilization medium containing 0-5 mM caffeine was examined. The sperm motility in the fertilization medium without caffeine was significantly higher than that in the fertilization medium with more than 2 mM caffeine. These results indicate that caffeine in the fertilization medium is not essential for bovine in vitro fertilization by fully capacitated spermatozoa, and that more than 2 mM caffeine has an adverse effect on preincubated (capacitated) sperm motility.

Animals↗

Annual summary of vital statistics--1998.

Most vital statistics indicators of the health of Americans were stable or showed modest improvements between 1997 and 1998. The preliminary birth rate in 1998 was 14.6 births per 1000 population, up slightly from the record low reported for 1997 (14.5). The fertility rate, births per 1000 women aged 15 to 44 years, increased 1% to 65.6 in 1998, compared with 65.0 in 1997. The 1998 increases, although modest, were the first since 1990, halting the steady decline in the number of births and birth and fertility rates in the 1990s. Fertility rates for total white, non-Hispanic white, and Native American women each increased from 1% to 2% in 1998. The fertility rate for black women declined 19% from 1990 to 1996, but has changed little since 1996. The rate for Hispanic women, which dropped 2%, was lower than in any year for which national data have been available. Birth rates for women 30 years or older continued to increase. The proportion of births to unmarried women remained about the same at one third. The birth rate for teen mothers declined again for the seventh consecutive year, and the use of timely prenatal care (82.8%) improved for the ninth consecutive year, especially for black (73.3%) and Hispanic (74.3%) mothers. The number and rate of multiple births continued their dramatic rise; the number of triplet and higher-order multiple births jumped 16% between 1996 and 1997, accounting, in part, for the slight increase in the percentage of low birth weight (LBW) births. LBW continued to increase from 1997 to 1998 to 7.6%. The infant mortality rate (IMR) was unchanged from 1997 to 1998 (7.2 per 1000 live births). The ratio of the IMR among black infants to that for white infants (2.4) remained the same in 1998 as in 1997. Racial differences in infant mortality remain a major public health concern. In 1997, 65% of all infant deaths occurred to the 7.5% of infants born LBW. Among all of the states, Maine, Massachusetts, and New Hampshire had the lowest IMRs. State-by-state differences in IMR reflect racial composition, the percentage LBW, and birth weight-specific neonatal mortality rate for each state. The United States continues to rank poorly in international comparisons of infant mortality. Expectation of life at birth increased slightly to 76.7 years for all gender and race groups combined. Death rates in the United States continue to decline, including a drop in mortality from human immunodeficiency virus. The age-adjusted death rate for suicide declined 6% in 1998; homicide declined 14%. Death rates for children from all major causes declined again in 1998. A large proportion of childhood deaths, however, continue to occur as a result of preventable injuries.

Adolescent↗

Adenosine triphosphate concentration and beta-D-glucuronidase activity as indicators of sea bass semen quality.

The most common parameters used to evaluate sperm quality are motility rate and duration and fertilization ability. In this study, chemical and biochemical parameters of sea bass (Dicentrarchus labrax) sperm were investigated to find an alternative method for evaluating sperm fertilization ability before and after cryopreservation. The biochemical and chemical analyses were performed with fresh and frozen-thawed sperm and seminal plasma. To cryopreserve sperm, 250-microl straws were used. Fertilization ability was evaluated by inseminating eggs (obtained from hormonally stimulated females) with fresh and cryopreserved sperm. The results revealed a linear relationship (P < 0.05) between semen fertilization capacity and some seminal plasma (beta-D-glucuronidase activity, potassium concentration) and sperm (ATP concentration, aspartate aminotransferase activity) parameters. Variations in semen fertilization rate could be best described by two multiple regression models: one including the sperm parameters and another including the seminal plasma parameters. For practical application, the use of simple regression models is of value. Fertilization rate in both fresh and cryopreserved sperm was reliably predicted by determining the ATP concentration or the beta-D-glucuronidase activity or both.

Adenosine Triphosphate↗

Comparing intracytoplasmic sperm injection and in vitro fertilization in patients with single oocyte retrieval.

OBJECTIVE: To compare the efficacy of intracytoplasmic sperm injection (ICSI) and conventional IVF in patients with favorable and poor sperm parameters in which only a single oocyte was available for insemination. DESIGN: Retrospective analysis. SETTING: University-affiliated IVF center. PATIENT(S): A total of 311 patients (425 cycles) who underwent either stimulated or spontaneous IVF cycles that resulted in single oocyte retrieval. INTERVENTION(S): The ICSI was indicated when fertilization failure was anticipated because of sperm quality or other confounding female factors. MAIN OUTCOME MEASURE(S): Fertilization rates and pregnancy rates (PRs) were analyzed according to the woman's age (< or =39 or >39 years), sperm quality, and mode of insemination. RESULT(S): In patients <39 years old with favorable semen quality, ICSI and standard insemination produced similar fertilization rates (67.1% vs. 75.0%) and PRs (0.0 vs. 8.2%). Conversely, in cases with apparent lower semen quality, ICSI gave a significantly higher fertilization rate (85.4% vs. 44.2%) but no significant difference in PRs (14.6% vs. 4.7%). In patients >39 years old and with favorable semen quality, ICSI and standard insemination produced similar fertilization rates (82.4% vs. 68.4%) and PRs (0.0 vs. 1.1%). The ICSI for lower semen quality, however, produced both higher fertilization rates (84.0% vs. 52%) and higher PRs (8.0% vs. 0). CONCLUSION(S): Our results suggest that in poorly responding patients, semen quality should remain the most important determinant when considering whether to perform ICSI. We have found that the values of 20 x 10(6)/mL and 35% motility are good predictors of success in such patients.

Adult↗

Intracytoplasmic sperm injection and conventional in vitro fertilization are complementary techniques in management of unexplained infertility.

PURPOSE: To evaluate the role of ICSI in unexplained infertility. METHODS: In 125 cycles with six or more oocytes retrieved per cycle, sibling oocytes were randomly allocated to IVF or ICSI (group A). In 74 cycles with less than six oocytes retrieved per cycle, cycles were allocated to IVF or ICSI (group B). RESULTS: In group A, ICSI fertilization rate of 61% per allocated oocyte was higher than IVF fertilization rate of 51.6% (P < 0.001). Complete fertilization failure occurred in 19.2 and 0.8% of cycles in IVF and ICSI, respectively (P < 0.001). In group B, fertilization rate in IVF cycles was 53.3% as compared to 60.7% per allocated oocyte in the ICSI cycles (P = 0.29). Complete fertilization failure was higher (P = 0.02) in conventional IVF (34.3%) than ICSI cycles (10.3%). CONCLUSIONS: Allocation of sibling oocytes to IVF and ICSI in the first cycle minimizes risk of fertilization failure. For patients with limited number of oocytes, ICSI technique is recommended.

Adult↗

Subzonal insemination (SUZI) or in vitro fertilization (IVF) in microdroplets for the treatment of male-factor infertility.

PURPOSE: The results of subzonal insemination (SUZI) and in vitro fertilization with microdroplet insemination used in couples with male-factor infertility are presented. RESULTS: The total fertilization rate was 17.4% for SUZI (n = 89) and 49.3% for microdroplet IVF (n = 100). The fertilization rate for standard IVF (n = 510), not including any male-factor infertility and performed during the same period, was 73.2%. The "take-home baby rate" per started cycle and per embryo transfer (ET), respectively, was 10 and 17.6% for SUZI and 20 and 24.7% for microdrop IVF. For standard IVF these figures were 27 and 31.7%. CONCLUSION: It was concluded that microdroplet IVF can be used with good results in cases of moderate male-factor infertility. The normal (2PN) fertilization rate with the SUZI technique was only 15.1%. However, despite the low fertilization rate, SUZI should be considered when dealing with severe male-factor infertility.

Fertilization in Vitro↗

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.

Adult↗

[The evaluation of 156 cycles of intracytoplasmic sperm injection in the management of male infertility and idiopathic infertility].

OBJECTIVE: The aim of this study was to examine, retrospectively, the efficacy of intracytoplasmic sperm injection (ICSI) in the treatment of male infertility and idiopathic infertility. METHODS: From November 1995 to January 1998, 140 couples (156 cycles) received ICSI because of severe male factor infertility or idiopathic infertility in our assisted reproductive technology center. The treatment cycles were divided into two groups, group I included 74 cycles done before May 1997, group II included 82 cycles done after May 1997. Mature oocytes were selected for microinjection. RESULTS: No differences were observed between the two groups in the number of mature oocyte and the total cleavage rate. Oocyte survival rate, normal fertilization rate, embryos of good morphology rate and clinical pregnancy rate were significantly higher in group II than that in group I. CONCLUSION: ICSI procedure does affect the quality of embryos. With the improvement of ICSI technique, ICSI should be the treatment of choice for male infertility and idiopathic infertility.

Adult↗

In-vitro fertilization in the presence of antisperm antibodies detected by the mixed antiglobulin reaction (MAR) and the tray agglutination test (TAT).

Data from 33 couples suffering from male immune infertility, who underwent 47 in-vitro fertilization (IVF) cycles between January 1989 and August 1991, were retrospectively analysed. The serum of all the 33 male partners had elevated tray agglutination test (TAT) titres (> or = 1:16) and positive mixed antiglobulin reaction (IgG MAR) test results in their semen. There was a slight correlation between these tests in semen and serum. Fertilization rates were analysed in three sperm MAR subcategories. Only the strongly positive MAR group (values > or = 90%) revealed a significant reduction in fertilization rate compared to the other MAR groups. However, this was not observed with increasing serum TAT titres. Fertilization rates were decreased in asthenozoospermic (20.1%) compared to normozoospermic (34.0%) male partners. This occurred also with couples not affected by immunological factors, but when antisperm antibodies were present, the fertilization rates were significantly poorer irrespective of whether the sperm motility was normal or decreased. Once fertilization occurred, the pregnancy rate was not affected by the severity of immunological factors. In assisted reproduction the sperm preparation techniques may reduce the inhibiting effects of antibodies bound to the spermatozoa, and when there are several oocytes to be inseminated, the chance of fertilization rises.

Adult↗

Comparison between PMSG- and FSH-induced superovulation for the generation of transgenic rats.

Superovulation protocols using single injections of pregnant mare's serum gonadotropin (PMSG) or minipumps with follicle-stimulating hormone (FSH) were compared in immature Sprague-Dawley (SD) rats. We used the following criteria: total number of ova, rate of fertilization, in vitro embryo development, sensitivity of zygotes to the microinjection of foreign DNA into the pronucleus, and their in-vivo development after transplantation into the oviduct of a recipient. Female SD rats were stimulated with 15 IU PMSG or 10 mg FSH followed by the injection of human chorionic gonadotropin (hCG) at doses of 20 and 30 IU per female. After hCG administration, they were mated with males of the same strain and sacrificed on day 1 of pregnancy. The percentage of mated animals and the fertilization rate was similar in all groups. In rats given PMSG, the number of ovulated zygotes was hCG dose-dependent. In contrast, the dose of hCG did not influence the efficiency of superovulation in rats given FSH, which was equal to PMSG-treated rats at the optimal dose of hCG. The rates of in vitro blastocyst development (31.4 and 23.3%) and the resistance to microinjection into the pronucleus did also not differ significantly between zygotes of both studied groups. The proportion of offspring developing from microinjected zygotes after oviduct transfer (26.2 and 26.8%, respectively) and the rate of transgene integration per newborns (7.3 and 4.9%, respectively) was similar in both experimental groups. The results of this study demonstrate that superovulation of immature SD rats by PMSG is equally effective as FSH treatment and, thus, preferable for transgenic rat technology due to the lower costs and easier handling.

Animals↗

Epididymal and testicular sperm for intracytoplasmic sperm injection in the treatment of obstructive azoospermia.

BACKGROUND: The possibility of treating male infertility because of obstructive azoospermia has been poor, but intracytoplasmic sperm injection (ICSI) has given this type of infertility sufferer a new option. MATERIAL AND METHODS: In this study 13 couples with obstructive azoospermia were treated in a total of 19 stimulated IVF cycles. The men were between 27 and 45 (mean 33) years of age. Their partners, 24-39 (mean 31) years of age were treated according to routine IVF procedures, i.e. down regulation with buserelin followed by hyperstimulation with urofollitropin. Fertilization was obtained by ICSI. Two embryos were transferred on day two after the ovum pick up. Sperm were retrieved through microsurgical epididymal aspiration (MESA) in four cycles, percutaneous epididymal sperm aspiration (PESA) in three cycles and through testicular sperm extraction (TESE) in 12 cycles. RESULTS: The overall fertilization rate was 68%, with a cleavage rate of 82%.The fertilization rate was equal (68%) with epididymal and testicular sperm and the cleavage rate was 87%) and 80%, respectively. Embryos were obtained for embryo transfer (ET) in all cases and five pregnancies (one twin pregnancy) were established (26% per ET), three using epididymal sperm and two using testicular sperm. CONCLUSION: Infertility due to obstructive azoospermia can successfully be treated with epididymal sperm and ICSI. When epididymal sperm cannot be found sperm extracted from a testicular biopsy can be used. PESA and TESE are quicker and easier alternatives to MESA and can be performed on an outpatient basis with local anesthesia.

Adult↗

Evaluation of sperm fertilizing ability using the Sperm Quality Analyzer.

The Sperm Quality Analyzer is an inexpensive device which provides a quantitative estimation of sperm motility. To evaluate the fertilizing ability of human spermatozoa using a Sperm Quality Analyzer, correlations amongst the sperm motility index, the sperm penetration index (as assessed using the sperm penetration assay; SPA), and the fertilization rate in the treatment of standard IVF-ET were analysed retrospectively. The sperm motility index demonstrated a significant correlation with sperm concentration (p < 0.001), sperm motility (p < 0.001) and the motile sperm concentration (p < 0.001) in a total of 104 fresh semen samples from 81 men donating samples for IVF-ET. The sperm motility index also showed a significant correlation (p < 0.001) with the sperm penetration index in 60 patients, assessed using the SPA, before they were treated by standard IVF-ET. The correlation between the sperm motility index and the IVF-ET fertilization rate was higher than that between the sperm penetration index and the fertilization rate. The sperm motility index was classified into three categories: 'poor' (sperm motility index < 80), 'medium' (sperm motility index 81-160) and 'good' (sperm motility index > 160). The relationships between the IVF-ET fertilization rate and each category of the sperm motility index values were also evaluated. For the three categories in the sperm motility index, the fertilization rates (76.0%) of 60 samples judged as 'good' were significantly higher than those (44.2%) of 15 samples judged as 'medium' (p < 0.001) and those (34.7%) of 13 samples judged as 'poor' (p < 0.001). These results indicate that the Sperm Quality Analyzer provides a reliable estimation of the fertilizing ability of human spermatozoa.

Embryo Transfer↗

Xenogenous fertilization of laboratory and domestic animals in the oviduct of the pseudopregnant rabbit.

Xenogenous fertilization was accomplished using bovine, porcine, and hamster follicular oocytes. The xenogenous fertilization rates for bovine and porcine follicular oocytes in the oviduct of the pseudopregnant rabbit were 13.4% and 2.0%, respectively. Temperatures of ovary, during transport to the laboratory, of 0 degrees or 37 degrees C had no effect on xenogenous fertilization rates of bovine oocytes. In vitro culture in 50 mug/ml FSH did not alter the xenogenous fertilization rates of bovine oocytes. Fertilization was observed with oocytes recovered 40 to 75 hr after insemination. Two cell embryos were recovered 70 to 75 hr after insemination. Ligation of the rabbit oviduct, number of ova deposited and sperm concentration did not affect the xenogenous fertilization rates of hamster ova. Cleavage of xenogenously fertilized hamster oocytes occurred between 28 and 29 hours after insemination.

Journal Article↗