Vascular endothelial growth factor, nitric oxide, and leptin follicular fluid levels correlate negatively with embryo quality in IVF patients

Objective(s): To measure vascular endothelial growth factor (VEGF), nitric oxide (NO) and leptin levels in individual ovarian follicles and to examine their relationships with perifollicular blood flow, follicular metabolic indices, and the developmental potential of the corresponding oocyte and embryo.

Design: Prospective study.

Setting: Academic, tertiary care institution.

Patient(s): Unselected IVF patients.

Intervention(s): Color-pulsed Doppler analysis of perifollicular blood flow; determination of partial pressure of oxygen (pO₂), partial pressure of carbon dioxide (pCO₂), and pH and VEGF, leptin and NO levels in follicular fluid.

Main Outcome Measure(s): Fertilization and day 3 embryo morphology and cleavage.

Result(s): Fifty-five follicular fluid samples from 16 patients were studied. Mean follicular fluid levels were as follows: VEGF, 1,046 ± 863.7 pg/mL (range, <63–3,332.7 pg/mL); NO₃/NO₂, 34.2 ± 12 μM (range, 16.4–76.1 μM); and leptin, 20.1 ± 12.1 ng/mL (range, 3.3–52.2 ng/mL). Vascular endothelial growth factor had a negative correlation with embryo morphology (r = −0.28, P=.01). Leptin demonstrated a negative correlation with follicular pO₂ (r = −0.42, P=.005) and a positive correlation with follicular pCO₂ (r = 0.36, P=.02). Follicular leptin levels correlated positively with VEGF levels (r = 0.46, P=.008) and with NO₃/NO₂ levels (r = 0.39, P=.006).

Conclusion(s): Vascular endothelial growth factor, NO and leptin appear to be markers of follicular hypoxia and suboptimal embryo development. Whether fluctuations of these regulatory factors determine or reflect changes in the follicular microenvironment affecting oocyte developmental potential remains to be elucidated.     

Hyaluronan in follicular fluids and fertilization of oocytes

Objective: To determine the concentrations of hyaluronan, E₂, and progesterone in follicular fluids (FFs) and the incidence of apoptotic granulosa cells. Also, to examine the relationship between the concentration of hyaluronan and follicular steroids, the incidence of apoptotic cells, and the fertilizability of the oocyte in the same follicle.

Design: Samples of 130 follicles were retrospectively analyzed for hyaluronan and steroids and the incidence of apoptotic cells.

Setting: The reproductive center in Yamagata University Hospital.

Patient(s): Forty women infertile because of tubal damage or unknown causes undergoing IVF treatment were selected.

Intervention(s): The samples were collected from follicle aspirations.

Main Outcome Measurement(s): The concentrations of hyaluronan and steroids in FFs, the incidence of apoptotic granulosa cells, and oocyte fertilizability.

Result(s): The levels of hyaluronan in FF were found to correlate positively with P (r=0.444, P<0.0001) and the incidence of apoptotic cumulus granulosa cells (r=0.387, P=0.002) and inversely with E₂ (r = −0.601, P<0.0001) and free T (r = −0.344, P=0.001). The concentration of hyaluronan in FFs containing a subsequently fertilized oocyte after insemination was significantly lower than that in FFs containing a subsequently unfertilized oocyte (P=0.0005) (fertilized, 50.0 ± 2.6 ng/mL; triploidy, 59.1 ± 6.8; and unfertilized, 66.9 ± 5.9).

Conclusion(s): The concentration of hyaluronan in FF is an indicator for estimation of oocyte viability for fertilization.     

Genetic contributions of the endothelial nitric oxide synthase gene to ovulation and menopause in a mouse model

OBJECTIVE: To investigate the influence of the endothelial nitric oxide synthase gene (Nos3) on ovulatory capacity and reproductive senescence. DESIGN: Prospective, controlled study. SETTING: Academic research institution. SUBJECT(s): Laboratory mice with targeted mutagenesis of Nos3. INTERVENTION(s): Hyperstimulation protocol, oocyte culture, and ovarian histology using wild-type (Nos3(+/+); n = 20), heterozygous (Nos3(+/m); n = 39), and homozygous deficient (Nos3(m/m); n = 11) female mice; observation of reproductive outcomes. MAIN OUTCOME MEASURE(s): Number and survival of oocytes; onset of menarche and menopause. RESULT(s): The mean number of superovulated oocytes (18 +/- 36 vs. 41 +/- 4) and the 48-hour overall survival rate of embryos (65% vs. 81%) were significantly reduced for Nos3(m/m) female mice compared with Nos3(+/+) female mice. Nos3(m/m) females showed a significantly reduced number and size of antral follicles and corpora lutea compared with wild-type controls. Compared with Nos3(+/m) x Nos3(+/m) breedings, Nos3(m/m) x Nos3(m/m) breedings showed a higher female age at first litter (76.2 +/- 10.3 vs. 107.8 +/- 26.6 days), fewer litters (10.5 +/- 3.6 vs. 7. 8 +/- 4.2), and a lower female age at reproductive senescence (400.2 +/- 64.5 vs. 332.1 +/- 27.4 days), respectively. CONCLUSION(s): Our data suggest that Nos3 deficiency is associated with reduced ovulatory capacity and impaired early embryonic viability and that it influences the onset of menarche and menopause.     

Dexamethasone compared with betamethasone for glucocorticoid treatment of postpartum HELLP syndrome

Objectives: To compare the efficacy of dexamethasone and betamethasone to ameliorate the course of postpartum hemolysis, elevated liver enzymes, low platelets (HELLP) syndrome. Methods: A prospective, mixed randomized/non-randomized clinical investigation of patients with postpartum HELLP syndrome. Treatment with either dexamethasone or betamethasone was continued until there was evidence of disease recovery. Results: Baseline characteristics of both the dexamethasone (n=18) and betamethasone (n=18) groups were similar. Although the time to discharge from the obstetrical recovery room was not statistically significant between groups, reduction in mean arterial blood pressure was more pronounced in the dexamethasone group as compared with the betamethasone group (−15.3±1.4 mmHg vs. −7.5±1.4 mmHg, respectively, P<0.01). Patients in the dexamethasone group required less antihypertensive treatment than the betamethasone group (6% vs. 50%, P=0.01) and also had a decreased need for readmission to the obstetrical recovery room (0% vs. 22%, P=0.03). Conclusion: This investigation supports the use of dexamethasone as the superior glucocorticoid to use for patients with postpartum HELLP syndrome.     

Ryudocan expression by luteinized granulosa cells is associated with the process of follicle atresia

Objective: To evaluate the presence of ryudocan in follicular fluid (FF) and its possible correlation with FF E₂ and P, and to study the levels of ryudocan in granulosa-lutein cells stimulated with hCG.

Design: Controlled clinical study and in vitro experiment.

Setting: University teaching hospital.

Patient(s): One hundred seven patients undergoing IVF.

Intervention(s): The FF and granulosa-lutein cells were aspirated from follicles 34 hours after an ovulatory gonadotropin bolus.

Main Outcome Measure(s): FF ryudocan, E₂, and P levels as well as hCG-mediated induction of ryudocan.

Result(s): Ryudocan was abundant in the FF; the concentration of ryudocan in human FF was estimated to be 305.5 ± 200.8 ng/mL (mean ± SD). Atretic follicles had higher concentrations of ryudocan (559.1 ± 156.5 ng/mL). FF ryudocan levels were inversely correlated with FF E₂ (r = −0.5023) and P concentrations (r = −0.4459). A detectable amount of ryudocan was found in pooled granulosa-lutein cells. Ryudocan production was augmented by surge levels of hCG.

Conclusion(s): Ryudocan is expressed in luteinized granulosa cells in vitro. The higher concentrations of ryudocan in FF of atretic follicles suggest an involvement of ryudocan in the process of atresia.     

The effects of clomiphene citrate on normally ovulatory women

Objective: To investigate the efficacy of clomiphene citrate (CC) on normally ovulatory women who complained of infertility.

Design: A randomized study.

Setting: University Hospital.

Patient(s): Thirty-three normally ovulatory women with unexplained infertility.

Intervention(s): Eighteen women received CC at a 50-mg dosage. Fifteen women received no ovulation-induction drugs.

Main Outcome Measures: The pregnancy rate (PR) per patient, the PR per cycle, and the cumulative pregnancy rate.

Result(s): Seven patients in the CC group stopped taking CC, and observations were terminated because of antiestrogenic effects. The pregnancy rate (PR) per patient and the PR per cycle were significantly decreased (P < 0.005) in the CC group (4 of 18 [22.2%] and 4 of 66 [6.1%], respectively) than in the spontaneous group (11 of 15 [73.3%] and 11 of 52 [21.2%], respectively). Kaplan-Meier tests showed that the cumulative pregnancy rate in the CC group was significantly lower than in the spontaneous group (P <0.05). Five of seven patients who had stopped taking CC became pregnant in spontaneous cycles.

Conclusion(s): Administration of CC to normally ovulatory women is not efficacious in terms of increasing the pregnancy rate.     

Effect of angiotensin-converting enzyme inhibitor on renal function in ovarian hyperstimulation syndrome in the rabbit

Objective: To investigate renal function and whether captopril prevents alterations in the handling of sodium and water in the ovarian hyperstimulation syndrome (OHSS) in the rabbit.

Design: Experimental study

Setting: Physiology laboratory.

Animal(s): Six female New Zealand white rabbits were used as controls, and 13 were hyperstimulated with gonadotropins.

Intervention(s): Saline or captopril.

Main Outcome Measure(s): Renal excretory and hemodynamic variables.

Result(s): The 3% extracellular volume expansion in OHSS animals induced a significant elevation in mean arterial pressure by 27%, although increments in natriuresis and diuresis were similar to those observed in controls. The OHSS group had impaired pressure-natriuresis sensitivity compared with controls (0.36 ± 0.07 μEq/min/g of Na excreted per mm Hg vs. 1.74 ± 0.45 μEq/min/g of Na excreted per mm Hg; P<.05. Captopril significantly reduced mean arterial pressure (P<.05) and shifted the pressure-natriuresis response to the left by 0.85 ± 0.17 μEq/min/g of Na excreted per mm Hg (P<.05).

Conclusion(s): In OHSS in the rabbit model, pressure-natriuresis sensitivity is impaired. Angiotensin II may play a significant role in this phenomenon, since angiotensin-converting enzyme inhibition normalized the pressure-natriuresis relationship.     

Comparison of the efficacy and safety of a highly purified human follicle-stimulating hormone (Bravelle) and recombinant follitropin-beta for in vitro fertilization: a prospective,randomized study

Objective: To compare the efficacy and safety of Bravelle s.c., Bravelle i.m., and Follistim s.c. in patients undergoing controlled ovarian hyperstimulation for IVF-ET.

Design: Open-label, randomized, parallel group, multicenter study.

Setting: Eleven academic and private fertility clinics with experience in IVF-ET.

Patient(s): Infertile premenopausal women with regular ovulatory menstrual cycles undergoing IVF-ET.

Intervention(s): Down-regulation with leuprolide acetate followed by up to 12 days of Bravelle s.c. (n = 60), Bravelle i.m. (n = 59), or Follistim s.c. (n = 58); hCG administration, oocyte retrieval, and ET.

Main Outcome Measure(s): Mean number of oocytes retrieved; patients with ET, chemical, clinical and continuing pregnancies; mean peak serum E₂ levels; adverse events and injection site pain scores.

Result(s): There were no significant differences among treatment groups in mean number of oocytes retrieved, peak serum E₂ levels, patients with ET, continuing pregnancies, or live births. There were no significant differences among the treatment groups in the number, nature, or intensity of adverse events. Patients treated with Bravelle s.c. or Bravelle i.m. experienced significantly less injection site pain than patients treated with Follistim s.c.

Conclusion(s): Bravelle s.c. and Bravelle i.m. are comparable in efficacy and safety to Follistim s.c. in patients undergoing controlled ovarian hyperstimulation for IVF-ET.     

Calcium to creatinine ratio in a spot sample of urine for early prediction of pre-eclampsia

Objectives: Based on the fact that urinary calcium excretion decreases in pre-eclampsia, this study was designed to determine the predictive value of calcium to creatinine ratio in a spot urine sample. Methods: The calcium to creatinine ratio was measured in a spot urine sample of 102 normotensive women at 20–24 weeks’ gestation who attended the prenatal care clinic of the Shiraz University of Medical Sciences. The women were followed-up until delivery and grouped according to pre-eclampsia occurrence. The prevalence of pre-eclampsia was measured and compared with the calcium to creatinine ratio. Results: Ninety-four women remained normotensive during pregnancy and eight developed pre-eclampsia. Mean age, gestational age at the beginning of the study, and gestational age at delivery did not differ significantly between the two groups. Mean urinary calcium concentration (15.9±8.5 mg/dl in normotensive vs. 10.2±7.5 mg/dl in pre-eclamptic women), and mean birth weight (3192±336.3 g vs. 2712±468.9 g) were significantly lower in pre-eclamptic patients (P=0.03 and 0.005, respectively). Mean calcium to creatinine ratio was also significantly lower in the pre-eclamptic group (P<0.03). Conclusions: Single urine calcium to creatinine ratio may be an effective method for screening women at greatest risk for pre-eclampsia.     

Increased angiotensin II in ascites during severe ovarian hyperstimulation syndrome: role of early pregnancy and ovarian gonadotropin stimulation

Objective: To investigate the implications of the ovarian renin-angiotensin system (RAS) in the pathophysiology of the ovarian hyperstimulation syndrome (OHSS) in relation to gonadotropin stimulation and early pregnancy.

Design: A controlled clinical study comparing blood and simultaneously sampled peritoneal fluid (PF) from patients with severe OHSS and from controls without OHSS.

Setting: University Hospitals.

Patient(s): Eleven patients with severe DHSS, 8 patients with ascites of other origin, 9 patients with a first-trimester pregnancy, and 15 patients stimulated with gonadotropins for IVF.

Main outcome measure(s): Angiotensin II immunoreactivity was measured in blood and PF and analyzed by high-performance liquid chromatography (HPLC) in ascites from OHSS.

Result(s): Angiotensin II immunoreactivity (pg/mL; mean ± SE) was highest in the ascites from pregnant OHSS (1,669 ± 418), reaching levels 5 times higher than in the plasma (331 ± 61) and 100 times higher than in control ascites (17 ± 6.7). Angiotensin II immunoreactivity was elevated in the PF during early pregnancy (211 ± 68) and after gonadotropin stimulation (244 ± 41) and was higher than in the plasma in both groups. Analysis by HPLC showed that the majority of Ang II immunoreactivity in the ascites of OHSS was because of true Ang II.

Conclusion(s): Severe forms of OHSS, especially those associated with pregnancy, are consistently characterized by huge concentrations of Ang II immunoreactivity in the ascites, proved to be true Ang II by HPLC analysis. This may be due to the synergistic effects of exogenous and endogenous hCG on the ovarian RAS.     

Decreased in vitro fertilization and cleavage rates after an equipment error during CO(2) calibration

Objective: To report the decreased IVF rate of oocytes and the reduced cleavage rate of pronuclear stage embryos after gas has backflowed from a gas analyzer into an incubator during the calibration of the CO₂ concentration.

Design: Case report.

Setting: Clinical research unit for reproductive medicine in a hospital.

Intervention(s): Backflow of gas from a gas analyzer into an incubator during calibration of the CO₂ concentration.

Main Outcome Measure: The IVF rate of oocytes and the cleavage rate of pronuclear stage embryos.

Result(s): Thirty-two oocytes were retrieved from four infertile female patients. Nine oocytes were fertilized and four fertilized oocytes were cleaved. In a related animal experiment, the cleavage rate was 57.1% (36/63) in the control group and 25.4% (16/63) in the study group (P = .00026).

Conclusion(s): The backflow of gas from the gas analyzer adversely affected the fertilization of oocytes and the cleavage of pronuclear stage embryos.     

Human oviductal cells reduce the incidence of apoptosis in cocultured mouse embryos

Objective: To investigate the effect of human oviductal cell coculture on the incidence of apoptosis in mouse embryos.

Design: Experimental laboratory study.

Setting: University gynecology unit.

Patient(s): Fallopian tubes were obtained from patients undergoing hysterectomy.

Intervention(s): Mouse embryos were cocultured with human oviductal cells.

Main Outcome Measure(s): Blastocyst development, allocation of inner cell mass (ICM) and trophectoderm (TE) in blastocyst, and apoptosis in embryos.

Results: Oviductal cells significantly enhanced the blastulation (38%) and hatching rate (22%) of the cocultured zygotes. The corresponding values in medium alone culture were 21% and 9%, respectively. The cocultured embryos also had higher blastomere count at blastocyst stage (P<0.005). This was due to increase in both the cell count of ICM (P<0.05) and TE (P<0.001). Coculture reduced the incidence of apoptosis in the cultured morula and blastocyst from 38% and 48% to 16% (P<0.001) and 27% (P<0.05), respectively. The number of apoptotic blastomeres per morula (1.5 ± 0.6; P<0.005) and blastocyst (2.3 ± 0.7; P<0.005) after coculture was also significantly lower than that of the corresponding control (morula, 2.1 ± 0.8; blastocyst, 3.5 ± 1.1).

Conclusion(s): Human oviductal cells improved mouse embryo development partly by decreasing the incidence of apoptosis.     

Bacteria in the transfer catheter tip influence the live-birth rate after in vitro fertilization

Objective: To assess the impact of individual bacteria isolated from the vagina and tip of the embryo transfer catheter on live-birth rates.

Design: Prospective clinical study.

Setting: Infertility outpatient clinic of a university hospital.

Patient(s): Ninety-one women undergoing IVF-ET.

Intervention(s): Cultures were obtained from the vagina for aerobic and anaerobic bacteria at the time of both sonographic egg retrieval and embryo transfer and from the tip of the embryo transfer catheter. Doxycycline treatment was started after egg retrieval.

Main Outcome Measure(s): The live birth of one or more neonates.

Result(s): Doxycycline had no substantial impact on the recovery of individual vaginal bacteria or on bacterial vaginosis. An increase in live-birth rate was associated with the recovery of hydrogen peroxide–producing Lactobacillus from the vagina (P=0.01) and from the embryo transfer catheter (P=0.01). In contrast, a reduction in live-birth rate was associated with recovery of Streptococcus viridans (S. viridans) from the embryo transfer catheter tip (P=0.04).

Conclusion(s): In the setting of IVF-ET, prophylactic doxycycline had little effect on vaginal bacteria. Specific bacteria recovered from the embryo transfer catheter appear associated with a detrimental or beneficial effect or with no effect on live-birth rates.     

Luteinizing hormone in peritoneal and ovarian cyst fluids: a predictor of ovarian carcinoma

Objective: The aim of this study was to define the role of luteinizing hormone (LH) as a tumor marker, specific for ovarian cancer. Methods: The study included 34 women with functional and benign ovarian cysts, 11 women with borderline ovarian tumors, 22 patients with advanced ovarian cancer and 15 patients with non-ovarian intraperitoneal malignancies. Serum, peritoneal fluid and ovarian cyst aspirates were obtained intraoperatively (laparoscopy or laparotomy) and were subjected to the LH analysis. Results: Peritoneal fluid LH levels were significantly increased in patients with ovarian cancer and those with borderline ovarian tumors as compared to patients with functional and benign ovarian cysts (P=0.005 and P=0.007, respectively). The patients with non-ovarian malignancies demonstrated the same peritoneal fluid LH levels as patients with benign ovarian tumors. There was no significant difference in the level of peritoneal fluid LH between ovarian cancer patients with and without ascites. The patients with functional and benign ovarian cysts demonstrated also significantly lower cyst fluid LH levels as compared to patients with malignant and borderline ovarian cysts (P=0.01 and P=0.03, respectively). Peritoneal and ovarian cyst fluid levels of LH were significantly increased in patients with fibrothecomas as compared to patients with other benign ovarian cysts. There were no significant differences in the serum LH levels comparing patients from all study groups. Conclusion: LH, detectable in peritoneal and ovarian cyst fluids, can be used as a tumor marker for identification of patients with borderline and malignant ovarian tumors.     

74例上皮性卵巢癌患者BRCA基因突变状况及临床意义分析

目的： 统计上皮性卵巢癌（EOC）患者的胚系乳腺癌易感基因（BRCA）致病突变情况,分析其与临床病理特征的关系。方法： 收集2017年1月—2020年1月在天津市中心妇产科医院诊治的EOC患者的胚系BRCA 基因检测情况,并对BRCA基因致病突变与患者临床病理资料进行分析。结果： 74例EOC患者中BRCA1突变18例（24.3%）,BRCA2突变7例（9.5%）,主要突变方式包括框移突变15例,无义突变5例,突变主要集中在外显子编码区（CDS9和CDS10）。BRCA基因突变组和BRCA基因野生组患者的临床分期、组织病理学类型、淋巴结转移、血清CA-125水平、恶性肿瘤家族史进行比较,差异均无统计学意义（P>0.05）。BRCA基因突变组患者无复发生存期（RFS）长于BRCA基因野生组,差异有统计学意义（Z=-1.931,P=0.027）;2组患者遗传性乳腺癌卵巢癌综合征家族史（HBOC家族史）率比较差异有统计学意义（χ² =4.059,P=0.044）。结论： 有HBOC家族史的EOC患者的家系女性成员进行BRCA基因检测是筛查卵巢癌高危患者的有效手段。     

Alternate pathway 17,20-lyase enzyme activity in the adrenals is enhanced in patients with polycystic ovary syndrome

Objective: To assess the activity of the enzyme cytochrome P450c17 in patients with polycystic ovary syndrome (PCOS).

Design: Prospective clinical study.

Setting: Outpatients at Erciyes University Medical School, Kayseri, Turkey.

Patient(s): Twenty-eight women with PCOS aged 25.44 ± 4.37 years (mean ± SD) and 18 normal women aged 26.94 ± 3.17 years.

Main Outcome Measure(s): Serum levels of 11-deoxycortisol, androstenedione (A), and 17-hydroxy progesterone were measured before and 30 and 60 minutes after ACTH (0.25 mg IV) injection.

Result(s): There was a statistically significant correlation between basal levels of 11-deoxycortisol (4.05 ± 1.16 ng/mL) and A (3.36 ± 0.97 ng/mL) (r = 0.539). The peak level of 11-deoxycortisol (7.82 ± 2.36 ng/mL) was also significantly correlated with the peak level of A (6.66 ± 1.32 ng/mL) (r = 0.570) in women with PCOS. There was no statistically significant correlation between basal A (2.33 ± 0.50 ng/mL) and basal 11-deoxycortisol (2.71 ± 0.59 ng/mL) or between peak A (3.38 ± 0.50 ng/mL) and peak 11-deoxycortisol (3.68 ± 0.48 ng/mL) levels in control subjects.

Conclusion(s): We believe that PCOS is characterized by enhanced activity of 17,20-lyase enzyme in an alternate pathway between 11-deoxycortisol and A in the adrenal glands.     

Alpha1B adrenoceptor expression is a marker of reduced survival and increased tumor recurrence in patients with endometrioid ovarian cancer

AIM: To investigate the expression patterns of different adrenoceptor isoforms in ovarian cancer and their association with survival and tumor recurrence. METHODS: The protein expression levels of α1B, α2C and β2 adrenoceptor were assessed in unselected ovarian cancer using immunohistochemistry on microarrayed archival tissue samples. A database containing clinical and pathology parameters and follow-up was used to investigate the association between adrenoceptor isoform expression with ovarian specific survival and tumor recurrence, using univariate and multivariate statistical analysis. RESULTS: Expression of α1B showed an association with reduced ovarian specific survival (P = 0.05; CI: 1.00-1.49) and increased tumor recurrence (P = 0.021, CI: 1.04-1.69) in the whole patient group. On sub-analysis the expression of α1B in endometrioid cancers (χ² = 5.867, P = 0.015) was found to predict reduced ovarian specific survival and increased tumor recurrence independently of tumor grade, clinical stage and chemotherapy. An association with clinical outcome was not seen for α2C or β2 AR. CONCLUSION: Alpha1B adrenoceptor protein was found to predict increased risk of tumor recurrence and reduced mortality in patients with endometrioid type ovarian cancer and should be investigated as a biomarker for identifying patients at increased risk of disease progression. Furthermore, α adrenergic receptor antagonists with α1B selectivity should be investigated as a possible adjuvant therapy for treating patients with endometrioid cancer. Proof of principle could be tested in a retrospective population study.     

Inducible and endothelial nitric oxide synthase: genetic background affects ovulation in mice.

OBJECTIVE: Inducible nitric oxide synthase (NOS) and endothelial NOS are involved in female reproductive physiology. We sought to investigate the influence of the inducible (Nos2) and endothelial (Nos3) NOS genes as a function of genetic background on ovulatory capacity and early embryonic development in a mouse model. DESIGN: Observational study of genetically altered mice and their response to a superovulation protocol. SETTING: Academic research institution. ANIMALS: Wild-type mice and mice deficient for Nos2 or Nos3 were bred to C57BL/6J and 129/Sv genetic backgrounds. INTERVENTION(S): Superovulation protocol, oocyte culture. MAIN OUTCOME MEASURE(S): Number of oocytes harvested, early embryonic development of zygotes, evaluation of ovarian histology. RESULT(S): The mean number of oocytes was significantly reduced in Nos3 deficient mice on a C57BL/6J background compared with controls. Oocytes deficient for Nos3 on a C57BL/6J background also showed reduced progression to two-cell stage embryos after 24 hours, two-cell stage embryos to blastocyst stage embryos, and survival to 48 hours. Those effects were distinctly absent in mice deficient for Nos3 on a 129/Sv background and in mice deficient for Nos2 on either genetic background. CONCLUSION(S): Our data show that disruption of Nos2 had no effect on ovulation in our mice. The negative effect of Nos3 deficiency on ovulatory capacity and early embryonic development is modulated by genetic background. This suggests a role for strain-specific modifier genes in these processes.     

Exogenous gonadotropin stimulation is associated with increases in serum androgen levels in in-vitro fertilization-embryo transfer cycles

Objective: To examine serum androgen profiles in women undergoing ovarian stimulation with exogenous gonadotropins in the setting of IVF-ET.

Design: Prospective study.

Setting: University hospital IVF-ET program.

Patient(s): Seventeen ovulatory women undergoing IVF-ET for endometriosis, male factor infertility, or tubal disease.

Intervention(s): A standard long protocol of GnRH agonist (GnRH-a) pretreatment (1 mg of leuprolide acetate SC for 10 days) was administered before ovulation induction with a urinary gonadotropin preparation.

Main Outcome Measure(s): After 10 days of GnRH-a treatment and on the day of hCG administration, serum concentrations of LH, T, androstenedione (A), sex hormone-binding globulin (SHBG), and DHEAS; the free androgen index (T/SHBG); and the number of follicles, oocytes, and embryos were assessed.

Result(s): Serum samples after 10 days of GnRH-a treatment showed incomplete LH suppression. While continuing the agonist during ovarian stimulation, LH values were suppressed further. However, serum T and A concentrations and the free androgen index showed a significant increase (samples drawn just before hCG administration). Serum T levels after 10 days of GnRH-a (before the administration of exogenous gonadotropins) were correlated negatively with the subsequent number of embryos.

Conclusion(s): Serum LH suppression with a conventional regimen of GnRH-a is incomplete in this heterogeneous group of ovulatory women. Exogenous gonadotropin stimulation results in a marked increase in ovarian androgen secretion.