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Measurement of urinary estrogen level with the urinary estrogen micrometry method in in vitro fertilization and embryo transfer therapy.

We have evaluated a new micrometering kit using hemagglutination inhibition reaction for determination of total estrogens in urine of normal fertile women and women undergoing in vitro fertilization and embryo transfer (IVF-ET). The micrometering kit was sensitive enough to monitor follicular maturation, and its technique was simple and rapid. Results were obtained in 2.5 hours. By the analysis of the individual estrogen levels in urine collected every 4 hours from 20 IVF-ET patients, the following results were found, with a few exceptions: once the level had exceeded 60 ng/ml, the onset of the luteinizing hormone surge was induced within the next 72 hours; when the level had been under 60 ng/ml, the onset was not induced within the next 24 hours

Adult↗

The serum concentration of estradiol after embryo transfer and the decline from preovulatory levels may influence the success of IVF treatment.

AIM: To establish the influence that serum estradiol concentrations prior to oocyte retrieval and 3 days after embryo transfer have on the establishment of in vitro fertilization (IVF) pregnancy. METHOD: Preovulatory (day-0) and luteal-phase (day-6) estradiol concentrations were retrospectively analyzed in 310 infertile patients, undertaking 1st-cycle conventional IVF or intracytoplasmic sperm injection. RESULTS: The IVF treatment success is significantly reduced in patients with an estradiol level <600 pg/ml and also when a rapid decline in luteal-phase estradiol concentrations from preovulatory concentrations (day 0:day 6 ratio) was exhibited. CONCLUSION: A day 0:day 6 estradiol ratio >5 and a serum estradiol concentration <600 pg/ml may adversely impact on the establishment of pregnancy in IVF.

Adult↗

Stochastic modeling of multiple ovulation and embryo transfer breeding schemes in small closed dairy cattle populations.

Genetic changes and genetic drift in three small closed dairy cattle populations were examined with a stochastic simulation model. Multiple ovulation and embryo transfer and AI techniques were simulated in three populations, two with 88 breeding females each and one with 352 breeding females. The selection goal was to maximize genetic improvement in milk yield. The reduction in genetic variation due to inbreeding and linkage disequilibrium was accounted for in the simulation. Strict restriction against inbred mating slowed genetic progress significantly in the small population but would not be consequential in the larger population. However, allowing inbred mating in the smaller population caused a rapid accumulation of inbreeding. Linkage disequilibrium was as important as inbreeding in reducing genetic variation. Genetic drift variance was much smaller in the larger population.

Animals↗

Influence of antiphospholipid antibodies on pregnancy outcome in women undergoing in vitro fertilization and embryo transfer.

PROBLEM: Antiphospholipid antibodies (APA) are thought to be involved in recurrent pregnancy loss. Therefore, we investigated the impact of APA on pregnancy outcome in women undergoing in vitro fertilization and embryo transfer (IVF-ET). METHOD OF STUDY: Blood samples taken from 54 Korean women referred for IVF were tested for the presence of APA, anticardiolipin antibody IgG and IgM and lupus anticoagulant. The standard gonadotropin-releasing hormone agonist long protocol was used for ovarian stimulation. RESULTS: Nine patients (16.7%) were positive and 45 (83.3%) were negative for APA. There were no significant differences between the two groups in clinical characteristics such as age, infertility duration, and response to controlled ovarian hyperstimulation. However, pregnancy outcome significantly differed between the two groups (p < 0.05). The APA positive group and APA negative group had abortion rates of 62.5% and 20.0%, respectively and delivery rates of 37.5% and 80.0%, respectively. CONCLUSION: The presence of APA in women undergoing IVF-ET was associated with a poor pregnancy outcome.

Adult↗

[In vitro fertilization and embryo transfer in a Deaconess Institution. Experience from the first years].

Eighty-one women suffering from occluded Fallopian tubes and 29 women with unexplained infertility were treated with in vitro fertilization and embryo transfer (IVF-ET). All procedures were carried out on outpatients. A total of 24 pregnancies were achieved in the group of patients with occluded tubes whereas eight became pregnant in the group of patients treated for unexplained infertility. From these results, it is expected that 30% of women suffering from occluded tubes or unexplained infertility can achieve pregnancy following two IVF-ET treatment cycles.

Adult↗

The cytology of vaginal, cervical and endometrial smears obtained at the time of embryo transfer during in vitro fertilization.

Seventy-four women enrolled in an in vitro fertilization (IVF) program had cytologic smears of the vagina, cervix and endometrium obtained at the time of embryo transfer (ET). Of these, 68 vaginal, 46 cervical and 25 endometrial smears were available for cytologic examination. Of the 68 vaginal smears, 4% showed a proliferative pattern, 40% were early secretory and 56% were advanced secretory. The 46 cervical smears demonstrated a delayed hormonal effect, with 70% showing a proliferative pattern, 23% early secretory and 7% advanced secretory cytology. Endometrial cells were obtained only when the Jones catheter, which has a side opening, was used. Twenty-two patients had both vaginal smears and suitable endometrial smears. Of these, 8 of the 9 patients with early secretory vaginal cytology had secretory endometrium while 10 of the 12 patients with mid-secretory vaginal cytology had secretory endometrium. The value of endometrial cytology in predicting conception following IVF-ET is unknown. It seems, however, that a good correlation exists between endometrial and vaginal cytology and that the latter may be of value as an additional, noninvasive tool for the evaluation of endometrial development.

Adult↗

Predictive value of plasma human chorionic gonadotrophin beta subunit in diagnosing ectopic pregnancy after in vitro fertilisation and embryo transfer.

A study was conducted aimed at establishing a range of plasma concentrations of the beta subunit of human chorionic gonadotrophin that might predict ectopic pregnancy after in vitro fertilisation and embryo transfer. From May 1984 to February 1986, 161 consecutive pregnancies at the Monash University in vitro fertilisation unit were analysed by determining plasma beta human chorionic gonadotrophin concentrations between two and 10 weeks after oocyte collection. Eighty eight ongoing singleton pregnancies, 25 multiple pregnancies, 27 first trimester spontaneous abortions, 12 anembryonic pregnancies, and nine ectopic pregnancies resulted from these conception cycles. When compared with values for ongoing singleton pregnancies two weeks after oocyte collection plasma beta human chorionic gonadotrophin concentrations in ectopic pregnancies were significantly lower (p less than 0.05; Wilcoxon rank sum test). Two weeks after oocyte collection all plasma beta human chorionic gonadotrophin concentrations in the set of ectopic pregnancies were below 30.6 IU/l, which corresponded to the lower quartile (25th percentile) of beta human chorionic gonadotrophin concentrations in ongoing singleton pregnancies. The beta human chorionic gonadotrophin concentration corresponding to the lower quartile of ongoing singleton pregnancies at each week of gestation was used to derive the predictive value of various statistics in detecting ectopic pregnancy in patients after in vitro fertilisation. The sensitivity, specificity, predictive value of a positive result, predictive value of a negative result, and efficiency of a single plasma beta human chorionic gonadotrophin concentration in predicting ectopic pregnancy were 100%, 68.1%, 16.7%, 100%, and 70%, respectively, two weeks after oocyte collection. These results suggest that a single determination of the plasma beta human chorionic gonadotrophin concentration beginning 14 days after oocyte collection is clinically useful in predicting the outcome of pregnancy achieved by in vitro fertilisation. Ectopic pregnancy after in vitro fertilisation is more likely when beta human chorionic gonadotrophin concentration is below the lower quartile of values in ongoing singleton pregnancies achieved by the technique.

Chorionic Gonadotropin↗

Evaluation of a cut-off value for normal sperm morphology using strict criteria to predict fertilization after conventional in-vitro fertilization and embryo transfer in asthenozoospermia.

The predictive value of sperm morphology evaluation using strict criteria (MEUSC) on fertilization and pregnancy rates was evaluated in 60 couples undergoing in-vitro fertilization (IVF) and embryo transfer. In all, 47 of the male partners had a progressive sperm motility < 50% with a density > 20 x 10(6)/ml (asthenozoospermia) and 13 had normozoospermia. MEUSC was performed on the same semen sample that was used for IVF on at least 100 spermatozoa after Papanicolaou staining at x 1250 magnification. Defects in the head, mid-piece and tail were counted separately, borderline forms were classified as abnormal and the teratozoospermia index was calculated for each sample. Of the spermatozoa in the asthenozoospermic group, 13.0% were judged as normal on MEUSC versus 19.8% in normozoospermia (P = 0.0013). The number of normal spermatozoa on MEUSC correlated with the progressive motility in asthenozoospermia (rho = 0.41, P = 0.0043). Defects in the mid-piece (P = 0.0004) and tail (P = 0.025) were more common, and the teratozoospermia index (P = 0.015) was higher in asthenozoospermic than normozoospermic samples. The parameters of MEUSC did not correlate with the fertilization rate. In asthenozoospermia, differences in fertilization rates were calculated for each cut-off value between 4 and 10% normal spermatozoa on MEUSC. A cut-off value of 6% gave the best statistical power. If > 6% of spermatozoa were normal, a median of 66.7% of ova were fertilized, compared with a median fertilization rate of 35.4% if < or = 6% of spermatozoa were normal (P = 0.022). The highest cut-off value still giving significant discrimination was 8% (normal MEUSC > 8%, median fertilization rate 66.7%; < or = 8%, median fertilization rate 35.7%, P = 0.028), and this may be used as the critical value for normal morphology in asthenozoospermia. It is concluded that despite a significant linear relationship between morphology using strict criteria and progressive motility, MEUSC is still useful as a predictor for fertilization in asthenozoospermia. In cases of asthenozoospermia with < or = 8% normal spermatozoa on MEUSC, micromanipulation should be considered.

Adult↗

The immunotherapy during in vitro fertilization and embryo transfer cycles in infertile patients with endometriosis.

OBJECTIVE: To investigate if the immunotherapy with corticosteroids would improve the pregnancy rate in infertile patients with endometriosis who undergo in vitro fertilization and embryo transfer (IVF-ET). METHODS: Forty-two infertile patients with endometriosis plus tubal factor and 87 pure tubal infertility patients who underwent IVF-ET in our unit were allocated randomly to the corticosteroid treatment group and the control group. RESULTS: The prevalence of autoantibodies (antinuclear antibody, lupus anticoagulant, anticardiolipin antibody, rheumatoid factor) was elevated significantly in patients with endometriosis plus tubal factor compared with pure tubal infertility patients (38.1% vs 2.3%). Twenty-one patients with endometriosis plus tubal factor underwent 54 cycles of IVF-ET, receiving corticosteroids. Forty-three patients with pure tubal factor underwent 81 cycles of IVF-ET, receiving corticosteroids. Twenty-one patients with endometriosis plus tubal factor who underwent 57 cycles of IVF-ET and 44 patients with pure tubal factor who underwent 84 cycles of IVF-ET served as controls, not receiving corticosteroids. In patients with endometriosis plus tubal factor, there was a significantly higher clinical pregnancy rate per cycle in the treatment group, with 42.6% (23/54) compared with 22.8% (13/57) in the control group but no differences between 2 groups in spontaneous abortion rate (21.7% vs 15.4%) and multiple pregnancy rate (17.4% vs 15.4%). In patients with pure tubal infertility, there were no significant differences between the treatment group and control group in clinical pregnancy rate (40.7% vs 34.5%), spontaneous abortion rate (12.1% vs 10.3%) or multiple pregnancy rate (18.2% vs 10.3%). In the endometriosis plus tubal infertility group with autoantibodies, the clinical pregnancy rate per cycle was significantly higher in the treatment group at 40.9% compared with 14.8% in the control group. In endometriosis plus tubal infertility group without autoantibodies, there was no significant difference between 2 groups with respect to the clinical pregnancy rate per cycle (43.8% vs 30.0%). CONCLUSIONS: This study suggests that immunotherapy with corticosteroids could improve the clinical pregnancy rate in endometriosis patients undergoing IVF-ET and may be more effective in patients with positive autoantibodies.

Adrenal Cortex Hormones↗

Transcription and translation in bovine nuclear transfer embryos.

The development of bovine embryos reconstructed by nuclear transfer (NT) is poor compared to that of embryos produced by in vitro fertilization. One reason for this could be incomplete reprogramming of the transferred nucleus. Therefore, with a view to optimizing the conditions for NT, the reprogramming of blastomere nuclei from 16- to 32-cell-stage in vitro-fertilized (IVF) embryos was investigated following NT by fusion of individual blastomeres with cytoplasts prepared from oocytes at two different stages of maturation. Heterogeneous RNA (hnRNA) production, nucleolar ultrastructure, and protein profiles of the NT embryos up to the 8-cell stage were analyzed. In all NT embryos analyzed for their hnRNA production (n = 133), [3H]uridine incorporation was higher at the 1-, 2-, and 4-cell stages than in control IVF embryos (n = 50). Ultrastructural examination of 11 NT embryos revealed evidence of transcriptional activity; fibrillar and granular components were seen in the nucleolus at the 1-cell stage. At the 2-, 4-, and 8-cell stages, fibrillar components were still evident but granular components had become scarce. The hnRNA synthesis, however, was not reflected in the one-dimensional electrophoretic patterns of protein production in the NT embryos (n = 56); these were largely similar to those of IVF embryos (n = 34) of corresponding stages. Thus, NT embryos made in this way do not behave like equivalent IVF embryos, suggesting that reprogramming of the transferred nucleus is absent or incomplete.

Animals↗

Intramuscular versus vaginal administration of progesterone for luteal phase support after in vitro fertilization and embryo transfer. A comparative randomized study.

A total of 156 patients were randomly treated with exogenous natural progesterone (intramuscularly, 50 mg/day) and vaginal gel (90 mg/day) P or nothing (Controls) from the day before embryo transfer (ET) for two weeks. In case of positive beta-HCG, the treatment was continued for 12 weeks. Plasma P and 17 beta-Estradiol concentrations were estimated and compared with the control not supplemented group. Both treatments were able to increase significantly the luteal plasmatic values of P versus controls. The ongoing pregnancy and the living birth rates per transfer were significantly higher in the patients supplemented with intramuscular P than in those treated with vaginal gel P. The intramuscular natural P appears the most suitable route of administration for luteal phase support in IVF-ET procedures.

Administration, Intravaginal↗

Culture factors affecting the success rate of in vitro fertilization and embryo transfer.

The development of one-cell mouse zygotes to the blastocyst stage in vitro has been used as a quality control for the media and handling procedures employed for human in vitro fertilization and embryo transfer (IVF/ET). One-cell mouse zygotes were placed in culture in medium containing bovine serum albumin. Aliquots of the same batch of medium containing female patients' homologous serum were used for the fertilization and culture of human oocytes. The following procedures were associated with high rates of mouse embryo development and human pregnancies following IVF/ET: adequate gassing and equilibration of the medium, double-rinsing of pipets and catheters used to handle embryos, use of a HEPES-buffered medium for manipulating embryos in the absence of an atmosphere containing 5% CO2, control of excessive temperature in the vicinity of the embryos, and ET using medium containing 50% patient's serum. The institution of these procedures gave more consistent pregnancy rates. However, there was no obvious association between fertilization and cleavage of human oocytes and the quality of the medium ascertained by the mouse embryo development test. In a continuing trial, we are comparing two culture media (modified Tyrode's and a medium formulated on the composition of human fallopian tube fluid [HTF]) and two culture techniques (culture in medium under oil in petri dishes and in loosely capped tubes). Significantly more mouse zygotes developed in HTF medium compared to Tyrode's medium. In a randomized 2 X 2 factorial trial with human IVF/ET, the highest pregnancy rate occurred when fertilization and culture were carried out in HTF medium under oil, but numbers are not yet sufficient to show any statistical difference between treatments.

Animals↗

Heterotopic pregnancies after in vitro fertilization and embryo transfer.

Seventeen cases of heterotopic pregnancies are reported among 1648 clinical pregnancies after in vitro fertilization. The high prevalence of tubal damage among IVF patients and the use of superovulation and multiple embryo transfer might predispose patients to the condition. Nine patients reported abdominal pain and vaginal bleeding, five patients did not have symptoms, and three had acute abdominal emergencies. Transvaginal ultrasonography was superior to transabdominal ultrasonography in the diagnosis of extrauterine pregnancies. The presence of an intrauterine gestation sac in a patient without symptoms should not exclude the diagnosis of a concomitant extrauterine pregnancy until the pelvis is carefully visualized. Early diagnoses of viable ectopic pregnancies before rupture abolishes mortality and morbidity and offers the chance of patient selection for conservative treatment. In two patients the extrauterine gestation sac was treated by transvaginal aspiration and injection of potassium chloride under ultrasonographic guidance. The outcome of the intrauterine pregnancy was favorable regardless of the method of treatment of the ectopic pregnancy.

Abdomen↗

Subpopulations of human granulosa-luteal cells obtained from gonadotropin- or gonadotropin-releasing hormone agonist/gonadotropin-treated follicles in in vitro fertilization-embryo transfer cycles.

PURPOSE: Our purpose was to find the differences in granulosa-luteal cells obtained from gonadotropin-versus gonadotropin-releasing hormone (GnRH) agonist/gonadotropin-treated follicles in in vitro fertilization-embryo transfer (IVF-ET) cycles. METHODS: Granulosa-luteal cells were obtained from 45 follicles of women undergoing IVF-ET with gonadotropin releasing hormone (GnRH) agonist and human menopausal gonadotropin (hMG) and from 45 follicles of women with hMG IVF-ET cycles. Subpopulations of granulosa-luteal cells were observed by computerized image analysis in which human chorionic gonadotropin (hCG) was localized using immunoperoxidase staining. RESULTS: The luteinized granulosa-luteal cells from hMG-treated follicles were larger than those from GnRH agonist/hMG-treated follicles. The hMG-treated follicles contained more hCG-stained cells, particularly those with cytoplasmic hCG localization. CONCLUSIONS: We found differences in morphometric characteristics and hCG localization in granulosa-luteal cells obtained from hMG-versus GnRH agonist/hMG-treated follicles. We presume that the results indicate the influence and importance of luteal-phase support on the clinical pregnancy rate in GnRH agonist/hMG-treated IVF-ET cycles.

Adult↗

High fecundity rates following in-vitro fertilization and embryo transfer in antiphospholipid antibody seropositive women treated with heparin and aspirin.

This study was undertaken to explore whether intervention with heparin and aspirin (H/A) in selected patients undergoing in-vitro fertilization (IVF) and embryo transfer could improve fecundity rates. Specifically, it explored the possibility that women diagnosed with organic pelvic disease who demonstrated antiphospholipid antibodies (APA) could benefit from H/A administration in a similar manner to that used in patients with recurrent pregnancy loss. We used an enzyme-linked immunosorbent assay for six different phospholipids to identify patients who expressed APA before they underwent IVF/embryo transfer. This study was confined to the first IVF/embryo transfer cycle that followed assessment of APA status and accordingly, the number of IVF/embryo transfer cycles corresponds with the number of patients treated. APA seropositive patients were treated with aspirin, 81 mg orally q.d., and heparin 5000 IU s.c. b.i.d., beginning on day 1 of controlled ovarian stimulation. The endpoint for success was a live birth or an ultrasound confirming fetal cardiac activity (a viable pregnancy). The prevalence of APA in patients diagnosed with organic pelvic disease (53%) was much higher than in those without female pathology (14%). The administration of H/A to APA seropositive patients significantly (P < 0.05) improved the viable pregnancy rate (49%) compared to the untreated APA seropositive group (16%). The viable pregnancy rate for APA seropositive women treated with H/A was also significantly (P < 0.001) higher than for untreated APA seronegative patients (27%). We conclude that all women undergoing IVF/embryo transfer should be tested for APA prior to initiating ovarian stimulation and those with APA seropositivity should be treated with H/A.

Adult↗

[Initial results with intratubal embryo transfer].

The intratubal transfer of pronucleus or early cleavage stage embryos (2-8-cell stage) is a new method in the treatment of human infertility. Following transvaginal sonographic oocyte retrieval in vitro fertilization is performed and the fertilized (pronucleus-stage) or embryos are transferred into the tube by laparoscopy. In comparison to GIFT (intratubal gamete transfer) fertilization under in vitro conditions offers the advantage that the success of the fertilization process can be examined. Therefore, this method can give important diagnostic information - especially in cases of poor sperm quality or unexplained infertility. After fertilization in vitro and transfer into the tube the embryone development occurs in the physiological milieu of the oviduct. Up to now the new procedure was performed in 40 patients with male factor or unexplained infertility. In 11 cases (28%) a pregnancy could be achieved. One abortion, but no ectopic pregnancy was observed. No difference in the pregnancy rates could be observed between the transfer of pronucleus stage and the early cleavage stage embryos. This method combines the advantage of IVF and GIFT and offers a successful procedure for the treatment of infertility.

Adult↗

[Intra-tubal embryo transfer. A new therapy in andrologic sterility].

The intratubal transfer of pronucleus or early cleavage stage embryos (2-8-cell stage) is a new method in the treatment of human infertility. Following transvaginal sonographic oocyte retrieval, in vitro fertilization is performed and the fertilized oocyte pronucleus-stage or embryos are transferred into the tube by laparoscopy. By comparison to GIFT (intratubal gamete transfer), fertilization under in vitro conditions offers the advantage that the success of the fertilization process can be examined. Therefore, this method can give important diagnostic information--especially in cases of poor sperm quality or unexplained infertility. After fertilization in vitro and transfer into the tube the embryonic development occurs in the physiological milieu of the oviduct. Up until the present, the new procedure was performed in 40 patients with male factor or unexplained infertility. In 11 cases (28%) a pregnancy could be achieved. One abortion, but no ectopic pregnancy was observed. No difference in the pregnancy rates could be observed between the transfer of pronucleus stage and the early cleavage stage embryos. This method combines the advantages of IVF and GIFT and offers a successful procedure for the treatment of infertility.

Adult↗

Influence of serum follicle stimulating hormone to luteinizing hormone ratio during buserelin acetate-induced pituitary desensitization on ovarian response to exogenous gonadotrophins in an in-vitro fertilization and embryo transfer programme.

We investigated the effect of endogenous gonadotrophins during pituitary desensitization with gonadotrophin-releasing hormone agonist (GnRHa) on ovarian responsiveness or the outcome of in-vitro fertilization (IVF) and embryo transfer. The results of 67 women who participated in the IFV programme at Nagasaki University Hospital, Japan, were analysed retrospectively. All women received GnRHa from the third day of menstrual cycle, and ovarian stimulation with exogenous gonadotrophins was initiated when the serum oestradiol concentration decreased to < 30 pg/ml. The serum follicle stimulating hormone (FSH)/luteinizing hormone (LH) ratio, rather than serum FSH or LH concentrations during GnRHa-induced pituitary desensitization, showed a significant positive correlation with age and the total dose of exogenous gonadotrophins. The FSH/LH ratio also showed a significant negative correlation with oestradiol response and the number of retrieved oocytes, and was significantly lower in pregnant women compared with the non-pregnant group during pituitary desensitization. Our results indicate that, even under pituitary desensitization with GnRHa, the serum FSH/LH ratio influences individual ovarian responsiveness and the state of the intra-ovarian hormonal environment. Our results suggest that the FSH/LH ratio may be a useful clinical predictor of the ovarian response to exogenous gonadotrophins under pituitary desensitization.

Adult↗