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Ovarian 11 beta-hydroxysteroid dehydrogenase: potential predictor of conception by in-vitro fertilisation and embryo transfer.

Cortisol is converted to the inactive glucocorticoid, cortisone, in several tissues by 11 beta-hydroxysteroid dehydrogenase (11 beta HSD). We have recently measured 11 beta HSD activity in cultured human granulosa-lutein cells recovered from patients undergoing in-vitro fertilisation and embryo transfer (IVF-ET). We now report an association between the outcome of IVF-ET and 11 beta HSD activity in these cells. Of the 64 patients studied, 32 had detectable 11 beta HSD activity and none became pregnant; whereas 76% of the remaining "11 beta HSD-negative" patients achieved pregnancies. Hence 11 beta HSD activity may predict the outcome of IVF-ET.

11-beta-Hydroxysteroid Dehydrogenases↗

Higher pregnancy rates after in vitro fertilization and embryo transfer in cases with sperm defects.

One hundred thirty-three couples were classified into four groups according to previous sperm analyses. These couples underwent 237 in vitro fertilization trials that led to 46 clinical pregnancies (19.4%/trial). Clinical pregnancy rate per laparoscopy of the group with male defects was significantly higher than that of the normospermic couples (25 versus 14%, P less than 0.05). The fertilization rate was significantly reduced only if two sperm anomalies were present (P less than 0.05). However, the mean number of embryos transferred did not differ significantly among the four groups, but the proportion of trials without transfer was increased with regard to the severity of sperm defect. Implantation rate per embryo in cases of triple transfer was significantly higher in the group with sperm anomalies as compared with the normospermic cases (18 versus 10%, P less than 0.02). Correction of the male problem through IVF treatment allows higher female fertility to be disclosed in these cases.

Adult↗

The role of endometrial and subendometrial vascularity measured by three-dimensional power Doppler ultrasound in the prediction of pregnancy during frozen-thawed embryo transfer cycles.

BACKGROUND: A good blood supply to the endometrium is usually considered as an essential requirement for implantation. OBJECTIVE: The aim of this study was to evaluate the role of endometrial and subendometrial vascularity in the prediction of pregnancy during frozen-thawed embryo transfer (FET) cycles. METHODS: Women undergoing FET in natural or clomiphene-induced cycles after the first stimulated IVF treatment were recruited. A three-dimensional (3D) ultrasound examination with power Doppler was performed 1 day after the LH surge to determine endometrial thickness, endometrial pattern, pulsatility index (PI) and resistance index (RI) of uterine vessels, endometrial volume, vascularization index, flow index and vascularization flow index of endometrial and subendometrial regions. RESULTS: Women in the pregnant group were significantly younger and used less gonadotrophins in their stimulated cycle. Endometrial thickness, endometrial volume, endometrial pattern, uterine PI, uterine RI, endometrial and subendometrial 3D power Doppler flow indices were similar between the nonpregnant and the pregnant groups. The age of women was the only predictive factor for pregnancy. Receiver operating characteristic curve analysis revealed that the area under the curve was around 0.5 for all ultrasound parameters for endometrial receptivity. CONCLUSION: Vascularity of endometrial and subendometrial layers measured by 3D power Doppler ultrasound is not a good predictor of pregnancy in FET cycles if measured at one time point only.

Adult↗

Leukaemia inhibitory factor (LIF) gene mutations in women with unexplained infertility and recurrent failure of implantation after IVF and embryo transfer.

OBJECTIVE: Leukaemia inhibitory factor (LIF) plays a central role in the control of implantation. We undertook this study to investigate the prevalence of LIF gene alterations in women with unexplained infertility and with recurrent failure of implantation after in vitro fertilisation (IVF) and embryo transfer. PATIENTS AND METHODS: Forty five women with recurrent failure of implantation after IVF (group A), 50 with unexplained infertility (group B) and 105 fertile women (controls) were screened for LIF gene mutations. Standard genomic DNA extraction, PCR amplification of the LIF gene and single-strand conformation polymorphism (SSCP) analysis were used to search for mutations which were subsequently confirmed by DNA sequencing. RESULTS: In group A, one woman was identified as having a neutral LIF gene polymorphism in exon 3 without affecting protein conformation. In group B, one woman with a heterozygous mutation and one with a neutral polymorphism were detected. In controls, only one woman with a neutral polymorphism in the intron between exons 2 and 3 was found. The woman with a potentially functional LIF gene mutation in group B achieved an ongoing clinical pregnancy after ovarian superovulation. DISCUSSION: Potentially functional mutations in the LIF gene do infrequently occur in women with unexplained infertility and may play a role in the etiology of infertility. However, routine screening for LIF mutations or polymorphisms in these women is not justified for the low prevalence of gene alterations.

Abortion, Habitual↗

Follicular fluid inhibin levels in relation to age in patients in an in vitro fertilization and embryo transfer programme.

Inhibin (INH), oestradiol (E2) and progesterone (P) were measured in the follicular fluid (FF) of 22 patients 28-38 years old (Group A) and 11 patients 43-47 years old (Group B) who had received gonadotrophin stimulation in an in vitro fertilization and embryo transfer (IVF-ET) programme. The results indicated that INH, E2 and P levels were significantly lower and the E2/P ratio was higher in FF of Group B patients (older women) compared to those of Group A. There were six single pregnancies among patients of Group A. No difference was observed in follicular fluid INH, E2 and P levels as well as in E2/P ratio between pregnant and non-pregnant patients of Group A (Group A1 and Group A2, respectively). A positive correlation was found between FF concentrations of E2 and P, E2 and INH and P and INH in the three Groups and a negative one between INH and the E2/P ratio in Group B. It seems likely that ovarian INH and E2 production are controlled by different mechanisms and that INH response to ovarian hyperstimulation is altered by advancing age.

Adult↗

Human chorionic gonadotrophin self-administered by the subcutaneous route to induce oocyte maturation in an in-vitro fertilization and embryo transfer programme.

This study was initiated to evaluate oocyte maturation and the outcome of in-vitro fertilization (IVF) cycles following the s.c. administration of human chorionic gonadotrophin (HCG) by the patient herself or her partner. A group of 104 women who entered our IVF embryo transfer programme were prospectively randomized to have 5000 IU or 10,000 IU HCG s.c. or i.m. The HCG was administered for induction of the final oocyte maturation in cycles with pituitary down-regulation with a gonadotrophin-releasing hormone agonist according to a long protocol and where ovarian stimulation had been achieved with pure follicle stimulating hormone. The mean concentration of HCG in serum 12 and 36 h after the HCG injection was significantly higher in the women receiving 5000 IU i.m. compared to the s.c. route. However, in women receiving 10,000 IU HCG there were no significant differences in the mean concentrations 12 and 36 h after the injection, irrespective of the route of administration. Furthermore, there were no significant differences in the relative numbers of retrieved mature oocytes between the groups. When comparing the clinical outcome in the different groups, no significant differences were found between those receiving 5000 IU or 10,000 IU HCG, i.m. or s.c. Our data indicate that HCG can be given s.c. without reducing the chance of retrieving a mature oocyte and that the clinical outcome with regard to pregnancies is not negatively affected.

Adult↗

[Immunohistochemical characterization of the inflammatory infiltrate in the human endometrium prior to in vitro fertilization and embryo transfer].

UNLABELLED: Infiltrating leukocyte population appears at the time of implantation in the endometrium of human and other mammals with hemochorial placentation. This leukocyte population is mainly composed of macrophages and typical granular lymphocytes CD56+ CD16-. 63 endometrial samples were studied using immunocytochemistry, before IVF and embryo transfer. We evaluated the sensibility for the assessment of immunological relevant infiltrating cells, of classical histopathologic examination compared with quantitation of leukocyte-common-antigen antibody immunostaining. 21 samples were thoroughly studied during proliferative phase. RESULTS: The sensibility of classical histologic analysis for the semi-quantitative assessment of leukocyte population, even when performed by an experienced histopathologist, is only 62%. When leukocyte population during proliferative phase is dense, it is mainly composed of macrophages and CD56+ CD16- lymphocytes. CONCLUSION: Immunological reaction already reported around the time of implantation may occur at other period of the menstrual cycle. Close mechanisms of the control and functions of this characteristic reaction remain to clarify.

Adult↗

Correlation between follicular fluid content and the results of in vitro fertilization and embryo transfer. I. Sex steroids.

The steroid content of 72 follicular fluids (FF) obtained from 42 women subjected to ovulation induction with the luteinizing hormone-releasing hormone analogue D-Trp6 and human menopausal gonadotropin was studied in terms of the evolution of in vitro fertilization (IVF) and embryo transfer (ET) results. The FF were classified into several categories based on oocyte evolution. Individual values of FF estrone and estradiol (E2), as well as androstenedione and testosterone could not be correlated with ET outcome. However, FF progesterone (P) levels for follicles leading to pregnancy were significantly lower when compared with those in the other categories. The correlation between the E2/P ratio and E2 permitted the definition of a band wherein IVF-ET outcome was successful and enabled the characterization of different functional follicular maturational states.

Androstenedione↗

Controlled ovarian hyperstimulation adversely affects implantation following in vitro fertilization-embryo transfer.

PURPOSE: Our purpose was to determine if controlled ovarian hyperstimulation adversely affects implantation. METHODS: A retrospective comparison of pregnancy rates (PRs) and implantation rates was made between oocyte recipients versus their donors, who shared half of the retrieved oocytes, and regular patients undergoing in vitro fertilization-embryo transfer (IVF-ET) who were not sharing eggs. RESULTS: Higher implantation rates (39.0 vs 22.5%; P < 0.05) were found in recipients compared to donors in the stimulated cycle. However, no differences were seen in PRs or implantation rates in frozen ET cycles. The data for standard IVF patients were almost-identical to those for donors. CONCLUSIONS: Superior implantation rates and PRs in oocyte recipients versus donors were not related to better oocyte quality for recipients because of egg sharing or to a better uterine environment because of similar results with frozen ET in all three groups. An adverse effect of the hyperstimulation regimen best explains the difference.

Adult↗

Blastocyst embryo transfer is associated with a sex-ratio imbalance in favor of male offspring.

OBJECTIVE: To evaluate the sex ratio of offspring born after blastocyst transfers. DESIGN: Retrospective data analysis. SETTING: A large assisted reproductive technology center. PATIENT(S): We included 1,284 offspring from 937 deliveries during the period August 2003-August 2005. INTERVENTION(S): Tabulation and statistical analysis of all births resulting from fresh IVF cycles. The sex of resulting offspring was compared in both day 3 and blastocyst transfers for all births and for singleton deliveries. In addition, the sex of children conceived with the use of autologous oocytes and donor oocytes was evaluated. MAIN OUTCOME MEASURE: Sex ratio of offspring born following embryo transfers (ETs) after day 3 of culture and sequential blastocycst culture. RESULT(S): The overall sex ratio was significantly shifted toward males when blastocyst transfers were performed. Blastocyst transfers with only the use of autologous oocytes resulted again in a significantly higher proportion of male offspring. An even greater proportional difference was encountered in singleton offspring from donor oocytes. However, significance was not reached because of the limited number of offspring in the subgroup. CONCLUSION(S): This is the first individual-center report of a significant sex-ratio imbalance after the sequential media culture of blastocysts. The large imbalance in singleton births associated with the use of donor oocytes, although not significant, is cautionary in regard to the use of elective single ETs. Observation and publication of phenomena such as the effects of extended culture on the sex ratio of live-borns will allow us a better understanding of early differences in sexual dimorphism of the embryo, and will allow us to counsel our patients more appropriately.

Cleavage Stage, Ovum↗

Ruptured tubo-ovarian abscess complicating transcervical cryopreserved embryo transfer.

OBJECTIVE: To present a rare infectious complication related to transcervical ET, without prior transvaginal puncture. DESIGN: Case report. SETTING: Hadassah University Hospital, IVF-ET unit. PATIENT: One patient undergoing cryopreserved-thawed ET. INTERVENTIONS: Artificial preparation of the endometrium with E2 and P, followed by transcervical intrauterine cryopreserved-thawed embryo transfer. RESULTS: After ET, severe pelvic inflammatory disease (PID) with ruptured tubo-ovarian abscess was diagnosed and treated. CONCLUSIONS: Severe PID including tubo-ovarian abscess formation should be considered a potential complication after ET, even without transvaginal oocyte aspiration.

Abscess↗

Involvement of phenotypes and serum levels of haptoglobin in the outcome of in vitro fertilization and embryo transfer.

Haptoglobin (Hp) possesses many functions in the immune system and angiogenesis as well as scavenging released hemoglobin, and thus it is speculated that Hp may also play a critical role in the reproductive process. To investigate the possible involvement of Hp in the outcome of in vitro fertilization and embryo transfer (IVF-ET), the serum Hp of 50 patients who underwent IVF-ET was analyzed by the Western blot method. The results showed a higher pregnancy rate in the patients with the Hp2-2 phenotype (38%) than in those with the Hp2-1 phenotype (16%) (p = 0.08), and the serum alpha 2 subunit levels in the patients who achieved pregnancy within four attempts at IVF-ET were higher than in the others (p < 0.05). The results of this study suggest that Hp is involved in the outcome of IVF-ET.

Adult↗

Comparison of four synchronization protocols for fixed-time bovine embryo transfer in Bos indicus x Bos taurus recipients.

The objective was to evaluate the effects of 400 IU of eCG given on Days 5 or 8 of an estrus synchronization protocol with progesterone-releasing intravaginal devices (PRID) and estradiol benzoate (EB), in recipients for fixed-time embryo transfer. A secondary objective was to determine the effects of injectable progesterone (given concurrent with EB treatment). Three-hundred-and-four crossbred Bos taurus x Bos indicus beef heifers were randomly assigned to one of four treatment groups (2 x 2 factorial design). At unknown stages of the estrous cycle (Day 0), all heifers received a progesterone-releasing intravaginal device (PRID), plus 2mg of EB i.m., with or without a concurrent treatment of 50mg of progesterone i.m. Heifers were further subdivided to receive 0.15 mg of d-cloprostenol (PGF) i.m. and 400 IU of eCG i.m. on Days 5 or 8. In all heifers, intravaginal devices were removed on Day 8 and 1mg of EB was given i.m. on Day 9 (Day 10 was arbitrarily considered the day of estrus). On Day 17, all heifers with >1 CL or a single CL with a diameter > or =18 mm (based on ultrasonographic examination), received an in vitro produced (IVP) embryo by non-surgical transfer. On Day 17, there was an effect of day of eCG administration on the number of CL (1.35 +/- 0.08 versus 1.13 +/- 0.04, for Day 5 versus Day 8, respectively; P = 0.02) and (in a subset of 154 heifers) mean (+/-S.E.M.) plasma progesterone concentrations (2.41 +/- 0.26 versus 1.74 +/- 0.19 ng/mL; P = 0.03). Although the proportion of recipients transferred/treated and pregnant/transferred did not differ among groups, the proportion of recipients pregnant/treated tended (P = 0.1) to be higher in heifers treated with eCG on Day 5 versus Day 8 (47.0% versus 40.7%, respectively). Progesterone treatment had no significant effect. In conclusion, treatment with eCG (and D-cloprostenol) on Day 5 significantly increased the number of CL and plasma progesterone concentrations and tended to increase pregnancy rates, although progesterone treatment had no significant effect.

Administration, Intravaginal↗

Effect of smoking on ovarian reserve and ovarian stimulation in in-vitro fertilization and embryo transfer.

The effect of cigarette smoking on ovarian reserve as measured by basal serum follicle stimulating hormone (FSH) concentrations, and by the response to a standard ovarian stimulation protocol, was examined retrospectively in 173 consecutive women (108 non-smokers and 65 smokers) undergoing in-vitro fertilization (IVF) and embryo transfer treatment. Women who smoked had a higher mean basal serum FSH concentration (P < or = 0.0001), in particular younger (<36 years) women, and required a statistically significantly higher mean dosage of gonadotrophins for ovarian stimulation than the non-smokers (48.1 +/- 15.6 versus 38.9 +/- 13.6 ampoules, 75 IU/ampoule; P < 0.0001). A lower mean number of oocytes was obtained in smokers than non-smokers (6.2 +/- 3.4 versus 11.1 +/- 6.3, oocytes P < or = 0.0001) and the rate of abandoned cycles (18.5 versus 8.5%) and total fertilization failure (18.5 versus 8.5%) was higher. The clinical pregnancy rate per cycle in smokers was 16.9% versus 21.3% in non-smokers but this was not statistically significant. In conclusion, cigarette smoking in women appears to significantly reduce their ovarian reserve and lead to poor response to ovarian stimulation at an earlier age.

Adult↗

Sex hormone-binding globulin, oestradiol-17 beta, progesterone and testosterone at stimulation and after embryo transfer during in-vitro fertilization.

Serum concentrations of sex hormone-binding globulin (SHBG), oestradiol-17 beta progesterone and testosterone were measured in 23 gonadotrophin-stimulated menstrual cycles and in the implantation period [days 11-19 after human chorionic gonadotrophin (HCG) injection] following in-vitro fertilization and embryo transfer. Nine cycles resulted in successful pregnancies, one pregnancy ended in spontaneous abortion (week 14) and 13 cycles were without conception. SHBG levels were significantly elevated above pretreatment values from day 3 after HCG injection onwards. A significant positive correlation was found between increments in SHBG (delta SHBG) during the luteal phase and oestradiol/testosterone ratios during the follicular and luteal phases. In the pregnant cycles a significant positive correlation was also found between delta SHBG during the implantation period and oestradiol/testosterone ratios during the luteal phase and the implantation period. Significant negative correlations were found between delta SHBG and testosterone during the luteal phase in pregnant and non-pregnant women as well as between delta SHBG during the period corresponding to implantation and testosterone during the luteal phase in non-pregnant cycles. The results may reflect a modulating action of the oestrogen/androgen balance upon SHBG levels in subjects with supraphysiological oestradiol levels, such as in stimulated cycles and in very early pregnancy.

Adult↗

Egg donation in an in vitro fertilization program: an alternative approach to cycle synchronization and timing of embryo transfer.

A new flexible protocol for the induction of recipient endometrial cycles is presented. For stimulation of endometrial growth, a fixed dose of conjugated estrogens, 3.75 mg/d was employed. The duration of the proliferative phase varied from 9 to 14 days, thus being adjusted to match the length of the follicular phase of the donor. Embryo transfer was performed on the fifth day of progesterone administration. Four term pregnancies resulted from 12 treatment cycles. In the conception cycles, the hormonal support was continued until the luteal placental shift occurred, regardless of gestational age.

Embryo Transfer↗

Delayed embryo transfer in an in-vitro fertilization programme: how to avoid working on Sunday.

It has previously been demonstrated that use of gonadotrophin releasing hormone analogues allows the administration of HCG to be delayed and makes it possible to avoid oocyte retrievals at weekends. In this study, we demonstrate that it is also possible to avoid embryo transfers on Sundays, by delaying transfers of Friday retrievals until Monday, without any apparent change in the results. This facility should improve the cost-effectiveness of assisted reproduction programmes.

Buserelin↗

Interleukin-1beta and interleukin-1alpha may affect the implantation rate of patients undergoing in vitro fertilization-embryo transfer.

OBJECTIVE: To investigate whether interleukin-1beta (IL-1beta) and interleukin-1alpha (IL-1alpha) affect the implantation rate of patients undergoing IVF-ET. DESIGN: Follicular fluid and serum were obtained on the day of hCG administration, the day of oocyte retrieval, and the day of embryo transfer. SETTING: Cellular immunology laboratory in a research institute, a high technology IVF unit in a medical center, and a university hospital. PATIENT(S): Thirty-three women who were undergoing IVF-ET. MAIN OUTCOME MEASURE(S): IL-1beta and IL-1alpha were measured by specific ELISA and their levels were correlated with the implantation rate. RESULT(S): Classification of IVF-ET patients according to their implantation rate revealed significantly higher amounts of follicular fluid IL-1beta in the implantation versus nonimplantation cycles (68.5+/-24.6 pg/mL versus 20.5+/-13.4 pg/mL); The difference between the level of IL-1alpha in the two groups was not statistically significant(11.6+/-5.1 pg/mL versus 7.3+/-1.9 pg/mL). In parallel, systemic FSH/hMG-dependent IL-1beta and IL-1alpha production was observed in implantation cycles but not in nonimplantation cycles. Statistically significant IL-1beta and IL-1alpha production was observed after administration of hCG. CONCLUSION(S): Gonadotropins used during IVF-ET induce local and systemic production of IL-1beta and IL-1alpha. In addition, the implantation rate for IVF-ET patients who have detectable serum concentrations of IL-1beta and IL-1beta on the day of hCG administration could be higher than the rate for IVF-ET patients who do not have detectable concentrations of these cytokines.

Adult↗