Does acupuncture the day of embryo transfer affect the clinical pregnancy rate? Systematic review and meta-analysis.

The effects of acupuncture on IVF outcomes is still unknown. We carried out a systematic review and meta-analysis of RCT to determine whether acupuncture performed at the time of ET improves outcomes. We searched Medline and Embase from January 1990 to June 2017, for the following terms): (acupuncture; acupuncture therapy) and (reproductive techniques, assisted; in vitro fertilization; embryo transfer). We selected RCT that compared acupuncture with sham acupuncture or no treatment. We included only trials in which acupuncture involved the insertion of needles into traditional meridian points. We evaluated the methodological quality of the trials using the Cochrane risk of bias tool. The measure of treatment effect was the pooled odds ratio of achieving a clinical pregnancy, ongoing pregnancy, or live birth for women in the acupuncture group compared with women in the control group. For pooled data, summary test statistics were calculated using the Mantel-Haenszel method, using the Rev-Man software, version 5.1. We analyzed six studies, including 2,376. In all trials, there were no significant differences between the groups concerning the mean numbers of embryos transferred, the mean age of the women undergoing the procedure, diagnose and use of ICSI. Acupuncture performed the day of ET was associated with a reduced risk of clinical pregnancy (0.87, 95% confidence interval 0.77 to 0.98). The pooled rate difference was -0.06 (-0.12 to -0.01) for clinical pregnancy. None of the trials reported significant adverse effects of acupuncture.

DHEA use to improve likelihood of IVF/ICSI success in patients with diminished ovarian reserve: A systematic review and meta-analysis.

The aim of this review is to determine if the use of DHEA increases the likelihood of success in patients with POR. We searched MEDLINE and EMBASE using the terms "DHEA and diminished ovarian reserve", "DHEA and poor response", "DHEA and premature ovarian aging". A fixed effects model was used and Peto's method to get the odds ratio (OR) with 95% confidence intervals (CI 95%). For quantitative variables, Cohen's method was used to present the standardized mean differences (SMD) with their corresponding confidence intervals. Only five studied fulfilled the selection criteria. DHEA was administered in 25 mg doses, three times a day. In all studies, the authors corrected for the presence of confounding variables such as partner's age, infertility diagnosis and number of transferred embryos. The meta-analysis of the five selected studies assessed a total of 910 patients, who underwent IVF/ICSI, of which 413 had received DHEA. DHEA use was associated with a significant increase in pregnancy likelihood (OR 1.8, CI 95% 1.29 to 2.51, p=0.001). When analyzing the association between DHEA use and the likelihood of abortion, we found low heterogeneity between studies (I2=0.0%) and the use of DHEA to be associated to a significant reduction in the likelihood of abortion (OR 0.25, CI 0.07 to 0.95; p=0.045). Analysis of the association of DHEA with average oocyte retrieval showed high variability between studies (I2=98.6%), as well as no association between DHEA use and the number of oocytes retrieved (SMD -0.01, CI 95% -0.16 to 0.13; p<0.05).

Is the risk of preeclampsia higher in donor oocyte pregnancies? A systematic review and meta-analysis.

OBJECTIVE: Preeclampsia (PE) occurs in 4.6% of pregnancies worldwide. The social phenomenon of increasing maternal age has raised the demand for donor oocytes. Egg donation has allowed women with poor ovarian reserve, premature ovarian failure, genetic disorders or surgical menopause to get pregnant. Recipients provide a unique model of immune response because of the differences in the genetic makeup of mothers and fetuses. In PE, immune tolerance may be impaired as a result of having non-autologous eggs implanted. Egg donation is a highly successful assisted reproductive technology, despite the significant number of issues arising from the implantation of non-autologous eggs. This study aimed to determine whether there is an association between egg donation and preeclampsia. METHODS: A systematic review of the literature available in PubMed and Google Scholar was carried out from January of 1995 to August of 2016 using the terms 'oocyte donation, preeclampsia', 'oocyte donation, in vitro fertilization, preeclampsia', 'oocyte donation, preeclampsia, outcomes pregnancies', 'oocyte donation, obstetric outcome.' Only six retrospective cohort studies met the selection criteria. RESULTS: The meta-analysis revealed a statistically significant association between egg donation and onset of preeclampsia (OR 4.50; 95% CI: 3.28-6.19; p<0.0001). CONCLUSION: Oocyte donation is associated with increased risk of preeclampsia in singleton pregnancies. Therefore, it is crucial to properly record and assess this finding when egg donation is the chosen assisted reproductive technology to attain pregnancy.

There is no association between the presence of anti-thyroid antibodies and increased reproductive loss in pregnant women after ART: a systematic review and meta-analysis.

Women submitted to ART treatments represent a select subgroup of individuals. Several studies have described the relationship between TAI and pregnancy outcomes as a result of ART, with contradictory results. The purpose of this systematic review was to determine the association between TAI and the risk of miscarriage in pregnancies resulting from ART. MEDLINE via PubMed, LILACS and Embase were searched for studies published in peer-reviewed journals from 1999 to 2017. The studies were summarized using the fixed effects model and the Peto's method to calculate RR in order to flesh out the association between TAI and spontaneous abortion. Only four papers were included in this systematic review and meta-analysis. Thirty-one miscarriages were observed in 210 clinical pregnancies of women with antithyroid antibodies; and 158 miscarriages were seen in 1,371 pregnancies without antithyroid antibodies. The meta-analysis failed to find an association between TAI and higher risk of reproductive loss, RR=0.94 95% confidence interval: 0.71-1.24; p=0.879. In conclusion, the presence of antithyroid antibodies was not associated with increased reproductive loss in patients submitted to ART treatments. It is our opinion that the presence of antithyroid antibodies should be considered as a secondary biomarker of autoimmune disease, rather than an actual cause of miscarriage in patients undergoing ART. Due to the small amount of evidence on the matter, the determination of TAI before the initiation of ART should be limited to research contexts.

Are children born from singleton pregnancies conceived by ICSI at increased risk for congenital malformations when compared to children conceived naturally? A systematic review and meta-analysis.

Since 1992, the development of intra-cytoplasmic sperm injection (ICSI) has allowed infertile couples and couples affected by severe male factor infertility in particular, many of which with a history of failed traditional IVF, to become parents. This has generated considerable controversy over the safety of the procedure for the offspring. This systematic review seeks to determine whether evidence indicates that the use of ICSI increases the risk of congenital malformation in children born from singleton pregnancies versus naturally conceived children. Twenty-one of the 104 publications listed in the literature search were included in the analysis. Observational studies reported mostly an increased risk for congenital malformation; the risk of congenital malformations is 7.1% in ICSI and 4.0% in the general population (OR 1.99 (95% CI [1.87 - 2.11]). However, attributing higher risk solely to ICSI might seem far-fetched, as in vitro and simulation procedures, patient diseases, and ICSI indication may also be associated with higher risk of malformation.

Do women offered assisted reproduction technologies have a higher incidence of gynecologic cancer? A systematic review and meta-analysis.

The last two decades have seen an increase in the number of women diagnosed with infertility. The consequent growth in the use of assisted reproductive technologies (ART) calls for the determination of its long-term effects, including the risk of cancer. Many studies have attempted to answer this question, albeit with contradictory results. This review aimed to assess whether assisted reproductive technologies are associated with an increased risk of gynecological cancer. A search for papers in the literature was carried out on MEDLINE, TRIP DATABASE and NICE, resulting in 11 studies enrolling 3,900,231 patients altogether. Of these, 118,320 were offered ART. The incidence of gynecological cancer in the group offered ART was 0.6%, while the incidence in the group not offered ART was 2.1%. Taking all the studies into consideration, women offered ART were not at greater risk of having gynecological cancer; instead, a protective association was found.

¿Mejora el estudio genético pre-implantacional la probabilidad de embarazo clínico en parejas sometidas a IVF/ICSI?: Revisión sistemática y meta análisis / Does the pre-implantation genetic study improve the probability of clinical pregnancy in couples undergoing IVF / ICSI?

Una de las principales causas de la falla de los ciclos de fecundación in vitro es la aneuploídia embrionaria. OBJETVO: determinar si las pruebas de tamizaje genético pre-implantacional favorece la posibilidad de embarazos MÉTODOS: Se realizó un revisión sistemática y meta-análisis. Buscamos en las bases PUBMED y EMBAS, estudios publicados entre 2006-2016, que compararan el número de embarazos en fecundación in vitro con y sin tamizeje genético preimplantaicon RESULTADOS: De los 115 artículos analizados, 4 cumplieron los criterios de selección. Evaluamos un total de 221 ciclos con tamizje y 592 sin. No encontramos diferencias en la posibilidad de embarazo (RR 0.88; IC95% 0.71-1.10; p=0.28). Al hacer análisis por subgrupo de técnica de tamizaje, encontramos que el uso de hibridación fluosrescente in situ se asoció a una disminución en la posibilidad de embarazo (3 estudios, RR 0.53; IC95% 0.36-0.77; p=0.0009); mientras que el uso de hibidración genómica comparativa se asoció a un aumeto (1 estudio, RR 1.58; IC 95% 1.24-2.00; p<0.001). CONCLUSION: La eficacia de las pruebas de tamizaje genñeticos son dependientes de la técnica, por lo que se deberia favorecer el uso de hibidración genómica comparativa.

Probably, the main cause in IVF failure is the transfer of aneuploid embryos. OBJECT: To determine if the use of preimplantational genetic screening improves the pregnancy rate in IVF cycles, compared to regular IVF. METHODS: We performed a systematic review and meta-analysis, searching in PUBMED and EMBASE databases studies published between 2006-2016, comparing the pregnancy rates in women undergoing IVF with PGS with that of women undergoing IVF only. RESULTS: Of the 115 articles found, 4 met the selection criteria, with a total of 734 women between 33 and 41 years: 221 with PGS and 592 controls. We found no association between the use of PGS and pregnancy (RR 0.88, 95% CI 0.71-1.10, p = 0.28). However, we performed a subgroup analysis by technique of PGS, and found that fluorescent in situ hybridization was associated with a diminished risk of pregnancy (3 studies;(RR 0.53; 95% CI 0.36-0.77; p = 0.0009), whereas comparative genomic hybridization was associated with an increase (1 study, RR 1.58, CI95% 1,24-2.00, p<0.001). CONCLUSION: The effectivity of PGS is determined by the technique for PGS; therefore, only comparative genomic hybridization should be offered.

Is there a reason to perform ICSI in the absence of male factor? Lessons from the Latin American Registry of ART.

STUDY QUESTION: Does the use of ICSI offer any outcome advantage over IVF in patients with non-male factor infertility? SUMMARY ANSWER: We did not find any outcome improvement that justifies the routine use of ICSI over IVF in non-male factor ART cycles. WHAT IS ALREADY KNOWN: Since its introduction in Latin America, the use of ICSI has increased substantially, even among patients without male factor infertility. However, it is not clear whether ICSI provides an advantage over IVF in non-male factor infertility. STUDY DESIGN SIZE DURATION: A retrospective cohort study of fresh cycles performed in 155 ART clinics located in 15 Latin American countries between 2012 and 2014. Records were assessed for 49,813 ART cycles (39,564 ICSI and 10,249 IVF) performed in infertile couples who did not have male factor infertility. Student's t-test was used to analyze normally distributed data, Wilcoxon test to analyze non-normally distributed data, and Fisher's exact test for categorical data. Logistic regression was used to quantify the effect of ICSI on delivery rate, adjusting for age of female partner, number of oocytes inseminated, number of embryos transferred, and transfer at blastocyst stage as possible confounding factors. Poisson regression analysis was used to quantify the effect of ICSI on fertilization rate, adjusting for age of female partner. PARTICIPANTS/MATERIALS SETTING METHOD: Cycles with the diagnosis of male factor and use of cryopreserved semen and with a freeze-all strategy were excluded. MAIN RESULTS AND THE ROLE OF CHANCE: After correcting for age of female partner, number of oocytes inseminated, number of embryos transferred and transfer at blastocyst stage, we found that the use of ICSI was associated with a significant decrease in the odds of delivery compared to IVF (odds ratio 0.88, 95% CI 0.84 to 0.93; P < 0.0001). LIMITATIONS REASONS FOR CAUTION: An important limitation of this study is the lack of randomization owing to its retrospective nature. This could result in selection bias, i.e. couples with the worst prognosis undergoing ICSI, or patients with a history of fertilization failure in IVF cycles undergoing ICSI. More than one cycle from the same couple may be included in the study. WIDER IMPLICATIONS OF THE FINDINGS: The lack of an outcome benefit-and, indeed, a reduced likelihood of delivery-following ICSI in non-male factor infertile couples suggests that ICSI may not be the most appropriate clinical approach in these patients. STUDY FUNDING/COMPETING INTERESTS: None.

[Cryopreservation in blastocyst stage effectively reduce the number of embryos cryopreserved]. / La criopreservación embrionaria en estadios más avanzados de desarrollo disminuye la frecuencia de pacientes con embriones criopreservados y el número de embriones criopreservados.

BACKGROUND: Multiple pregnancies are the main complication associated to assisted reproduction, due to the transfer of more than one embryo. Embryo cryopreservation allows the sequential transfer of all generated embryos, thus diminishing the risk of multiple pregnancies. However, it leads to accumulation of cryopreserved embryos. To reduce their accumulation in our unit, we started to preserve embryos as blastocysts, that have a rate of successful pregnancies of approximately 40%. AIM: To perform a sensitivity analysis of this change of policy on the accumulation of embryos. MATERIAL AND METHODS: Records of 571 cycles of in vitro fertilization since 2007 were reviewed, assuming a transference rate of two embryos in women aged less than 35 years and three embryos in older women. The number of embryos that would be preserved as zygotes, eight cell stage or blastocysts, was analyzed. RESULTS: Multiple component logistic regression analysis showed a 20% reduction in the odds ratio of cryopreservation per year of age. There was a 95% reduction in the ratio, when comparing the preservation of third and first day embryos and a 99% reduction when comparing preservation of embryos in blastocyst stage and first day embryos. CONCLUSIONS: Cryopreservation at blastocyst stage effectively decreased the frequency of embryo cryopreservation.

Decreased anti-Müllerian hormone concentration in follicular fluid of female smokers undergoing artificial reproductive techniques.

BACKGROUND: Several reports indicate that women who smoke have an increased risk of failure to conceive compared with their non-smoker counterparts. Here, we assessed the effect of smoking during the Assisted Reproduction Therapy (ART) on a potential marker of ovarian reserve, anti-müllerian hormone (AMH) in the follicular fluid (FF). MATERIALS AND METHODS: This was a cohort prospective study to assess the association between cigarette smoking and AMH concentrations in FF in fifty-six women undergoing their first ART cycle. Self-reported smoking status over time was also collected through personal interview. The main outcome measured was the association between current smoking and AMH concentrations in FF. Smoking status was assessed by FF cotinine concentrations. Analysis of covariance was performed to test statistical interaction between the main outcome and confounders. RESULTS: The mean concentration of AMH in follicular fluid was significantly decreased among smokers (1.02±0.14 vs. 1.74±0.15, P<0.05). No statistical interaction was found between this difference in AMH concentrations and confounders like age and BMI. Thus, our data support the idea that AMH is decreased in active smokers across the fertile age. CONCLUSIONS: The hypothesis of decreased AMH concentration in follicular fluid in female smokers was confirmed. The mechanisms through which cigarette smoking induces this fall in AMH are unknown and additional research is needed to improve our comprehension of the negative impact of smoking on ART outcomes.

La criopreservación embrionaria en estadios más avanzados de desarrollo disminuye la frecuencia de pacientes con embriones criopreservados y el número de embriones criopreservados / Cryopreservation in blastocyst stage effectively reduce the number of embryos cryopreserved

Background: Multiple pregnancies are the main complication associated to assisted reproduction, due to the transfer of more than one embryo. Embryo cryopre-servation allows the sequential transfer of all generated embryos, thus diminishing the risk of multiple pregnancies. However, it leads to accumulation of cryopreserved embryos. To reduce their accumulation in our unit, we started to preserve embryos as blastocysts, that have a rate of successful pregnancies of approximately 40%. Aim: To perform a sensitivity analysis of this change of policy on the accumulation of embryos. Material and Methods: Records of 571 cycles of in vitro fertilization since 2007 were reviewed, assuming a transference rate of two embryos in women aged less than 35 years and three embryos in older women. The number of embryos that would be preserved as zygotes, eight cell stage or blastocysts, was analyzed. Results: Multiple component logistic regression analysis showed a 20% reduction in the odds ratio of cryopreservation per year of age. There was a 95% reduction in the ratio, when comparing the preservation of third and first day embryos and a 99% reduction when comparing preservation of embryos in blastocyst stage and first day embryos. Conclusions: Cryopreservation at blastocyst stage effectively decreased the frequency of embryo cryopreservation.

[Live birth after the transfer of embryos obtained by fertilization of vitrified oocytes: report of one case]. / Parto tras la transferencia de embriones generados por fecundación de ovocitos vitrificados: Comunicación de un caso.

We report the first successful live birth after the transfer of embryos obtained by fertilization of vitrified oocytes. A couple with primary infertility due to teratozoospermy and chronic an ovulation decided to undergo assisted reproductive technology after 4 failed cycles of super ovulation and intrauterine insemination. The woman underwent a standard luteal phase agonist protocol, with controlled ovarian hyper stimulation with daily 150 IU recombinant gonadotropin and 75 IU urinary gonadotropin for 12 days. Due to the high risk of ovarian hyper stimulation syndrome the couple was advised to delay embryo transfer. Eighteen mature oocytes were recovered, eight were vitrified and 10 fertilized and cryopreserved as pro-nuclei. Two months later, four vitrified oocytes were thawed, and three morphologically-normal embryos were transferred to an estrogen-progesterone-primed uterus, obtaining triple clinical pregnancy. The pregnancy was uneventful until 32 weeks of amenorrhea, when pre-term delivery started. After inducing lung maturity, a cesarean section was performed and three healthy fetuses were delivered. Nowadays, the babies are two and a half years old, and have had a normal psycho-motor development.

Parto tras la transferencia de embriones generados por fecundación de ovocitos vitrificados: Comunicación de un caso / Live birth after the transfer of embryos obtained by fertilization of vitrified oocytes: Report of one case

We report the first successful live birth after the transfer of embryos obtained by fertilization of vitrified oocytes. A couple with primary infertility due to teratozoospermy and chronic an ovulation decided to undergo assisted reproductive technology after 4 failed cycles of super ovulation and intrauterine insemination. The woman underwent a standard luteal phase agonist protocol, with controlled ovarian hyper stimulation with daily 150IU recombinant gonadotropin and 75IU urinary gonadotropin for 12 days. Due to the high risk of ovarian hyper stimulation syndrome the couple was advised to delay embryo transfer. Eighteen mature oocytes were recovered, eight were vitrified and 10 fertilized and cryopreserved as pro-nuclei. Two months later, four vitrified oocytes were thawed, and three morphologically-normal embryos were transferred to an estrogen-progesterone-primed uterus, obtaining triple clinical pregnancy. The pregnancy was uneventful until 32 weeks of amenorrhea, when pre-term delivery started. After inducing lung maturity, a cesarean section was performed and three healthy fetuses were delivered. Nowadays, the babies are two and a half years old, and have had a normal psycho-motor development.

Recent cigarette smoking and assisted reproductive technologies outcome.

OBJECTIVE: To assess the association between recent cigarette smoking (CS) in female and male partners and assisted reproduction technology (ART) outcomes. DESIGN: Cohort prospective study. SETTING: University ART program in Chile. PATIENT(S): One hundred sixty-six couples seeking pregnancy through ART. INTERVENTION(S): Follicular fluid (FF) and serum cotinine concentrations were measured in female partners. Self-reported CS data were collected through personal interviews. MAIN OUTCOME MEASURE(S): The association between female recent smoking, assessed by FF and serum cotinine concentrations, and ART outcomes, such as number of ova retrieved and implantation rates, and the association between self-reported male recent smoking and live birth rates. RESULT(S): A significant age-adjusted association between increased FF cotinine level and decreased number of ova retrieved was found. The male partner's smoking habit significantly decreased the live birth rate from 21.1% to 7.8%. Serum cotinine concentrations paralleled those of FF. CONCLUSION(S): The hypothesis of a detrimental effect of recent female smoking over implantation rates is rejected. However, recent male smoking is associated with significantly decreased live birth rates even after adjusting for confounders. Female recent smoking was significantly associated with decreased number of retrieved ova.

Cumulus cell apoptosis changes with exposure to spermatozoa and pathologies involved in infertility.

OBJECTIVE: To determine whether the incidence of apoptosis in mature oocyte cumulus cells changes after insemination related to infertility. DESIGN: Prospective study. SETTING: Public hospital and university. PATIENT(S): One hundred women undergoing in vitro fertilization and embryo transfer (IVF-ET). INTERVENTION(S): Collection of cumulus cells from IVF-ET cycles with different infertility etiologies. MAIN OUTCOME MEASURE(S): Detection of apoptosis in cumulus cells; fertilization, embryo quality, and pregnancy rate. RESULT(S): The incubation of cumulus-oocyte-complexes with spermatozoa led to an increase in cumulus cell apoptosis from 34.2 +/- 3.7 to 44.5 +/- 6.3%. After insemination, cumulus cells of poor quality embryos showed a statistically higher apoptotic rate versus cumulus cells of good quality embryos (61.5 +/- 6.4 vs. 40.6 +/- 3.9%). Cumulus cells arising from oocytes with >or=50% fertilization rates after insemination showed higher apoptosis rates did cumulus cells from oocytes with <50% fertilization rates (46.0 +/- 3.7 vs. 33.8 +/- 4.0%). Patients with endometriosis presented higher apoptotic rates before insemination (77.6 +/- 9.06%). Cumulus cells obtained after aspiration showed no differences in their apoptosis rates for the following factors: age of women, aspirated oocytes, estradiol level, fertilization rate, and embryo quality or pregnancy. The apoptotic profile from pregnant women was less than (but not statistically significantly different from) profiles from nonpregnant women. CONCLUSION(S): These results suggest that the incidence of apoptosis in cumulus cells is associated with exposure to spermatozoa and the cause of infertility.

The human corpus luteum: life cycle and function in natural cycles.

OBJECTIVE: To summarize recent advances in the understanding of the endocrine signaling pathways between the hypothalamus, pituitary, and human corpus luteum (CL); to examine the major paracrine and autocrine mechanisms and the key genes and proteins involved in CL development, function, and regression in natural cycles; to review the endocrine and molecular response of the midluteal phase CL to in vivo administration of human chorionic gonadotropin (hCG); and to describe the ultrasonographic and Doppler evaluation of the ovary and endometrium throughout the luteal phase. DESIGN: Published data in the literature, including the basic and clinical research studies of the authors. SETTING: University-affiliated hospital and research centers. PATIENT(S): None. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Clinical and molecular analysis of human CL function. RESULT(S): The endocrine function of the subpopulations of luteal cells is critical for the maintenance of CL function, including neovacularization and steroid hormones production. We consider the key genes and proteins that favor development of luteal structure and function throughout the menstrual cycle and in our model of hCG treatment resembling early pregnancy. CONCLUSION(S): These data indicate that the functional lifespan of the CL depends on paracrine and autocrine mechanisms. Therefore, the significance of the key genes and proteins that we analyze in lutein cells during CL development, function, demise, and rescue by hCG is likely to bring new therapeutic applications for the management of fertility defects and the control of fertility.

Differences in the endometrial transcript profile during the receptive period between women who were refractory to implantation and those who achieved pregnancy.

BACKGROUND: Gene expression profiling of normal receptive endometrium has been characterized, but intrinsic defects in endometrial gene expression associated with implantation failure have not been reported. METHODS: Women who had previously participated as recipients in oocyte donation cycles and repeatedly exhibited implantation failure (Group A, study group) or had at least one successful cycle (Group B, control group) and spontaneously fertile women (Group C, normal fertility group) were recruited. All were treated with exogenous estradiol and progesterone to induce an endometrial cycle, and an endometrial biopsy was taken on the seventh day of progesterone administration. RNA from each sample was analysed by cDNA microarrays to identify differentially expressed genes between groups. RESULTS: 63 transcripts were differentially expressed (>or=2-fold) between Groups A and B, of which 16 were subjected to real time RT-PCR. Eleven of these were significantly decreased in Group A with regard to Groups B and C. Among the dysregulated genes were MMP-7, CXCR4, PAEP and C4BPA. CONCLUSIONS: Repeated implantation failure in some oocyte recipients is associated with an intrinsic defect in the expression of multiple genes in their endometrium. Significantly decreased levels of several transcripts in endometria without manifest abnormalities is demonstrated for the first time and shown to be associated with implantation failure.

Tumor necrosis factor-alpha activates nuclear factor-kappaB but does not regulate progesterone production in cultured human granulosa luteal cells.

BACKGROUND: The role of tumor necrosis factor-alpha (TNF-alpha) in granulosa luteal cell function and steroidogenesis is still controversial. Our aim was to examine the steroidogenic response, together with the simultaneous expression and activation of nuclear factor-kappaB (NF-kappaB), in cultured human granulosa luteal cells (GLCs) following administration of TNF-alpha. MATERIALS AND METHODS: This prospective controlled study was conducted in the Human Reproduction Division at the Institute of Maternal and Child Research, Faculty of Medicine, University of Chile and the San Borja Arriarán Hospital, National Health Service, Santiago, Chile. GLCs were obtained from aspirates of follicles from women undergoing in vitro fertilization (IVF). Thirty-two women undergoing IVF for tubal-factor and/or male-factor infertility participated in this study. Protein levels of NF-kappaB, the NF-kappaB inhibitor IkappaBalpha and steroidogenic acute regulatory protein (StAR) were determined by Western blot and localization of NF-kappaB was studied by indirect immunofluorescence. Progesterone production was determined by radioimmunoassay. RESULTS: TNF-alpha did not affect the expression of StAR protein or the synthesis of progesterone. NF-kappaB was expressed in the GLCs and activated by TNF-alpha, resulting in degradation of IkappaBalpha and mobilization of the p65 NF-kappaB subunit into the nucleus. CONCLUSIONS: These results indicate that TNF-alpha did not modulate steroidogenesis in cultured human GLCs. However, NF-kappaB was activated by TNF-alpha. Therefore the activation of NF-kappaB via the TNF-alpha pathway is likely associated with other preovulatory granulosa cell processes important for human ovarian function.

AZFc partial deletions in Chilean men with severe spermatogenic failure.

OBJECTIVE: To determine the prevalence of AZFc subdeletions in infertile Chilean men with severe spermatogenic impairment. DESIGN: Prospective analysis. SETTING: University infertility clinic. PATIENT(S): Ninety-five secretory azo/oligozoospermic men without AZFc Y chromosome microdeletions: 71 whose testicular histology showed severe spermatogenic impairment and 24 who exhibited reduced testicular volume and elevated serum FSH levels. As controls, we studied 77 men (50 fertile and/or normozoospermic, and 27 with azoospermia and normal spermatogenesis). INTERVENTION(S): Peripheral blood was drawn to obtain genomic DNA for polymerase chain reaction (PCR) digestion assays of DAZ-sequence nucleotide variants and for AZFc-STS PCR after a complete testicular characterization (biopsy, hormonal, and physical evaluation). MAIN OUTCOME MEASURE(S): DAZ genes and AZFc subdeletion types. RESULT(S): In cases we observed two "gr/gr" subdeletions (2.1%), one with absence of DAZ1/DAZ2 (g1/g2 subtype), and the other with absence of DAZ3/DAZ4 (r2/r4 subtype). Additionally, we found a g1/g3 subdeletion in a patient with Sertoli-cell-only syndrome. In controls, we observed two gr/gr subdeletions with absence of DAZ1/DAZ2 (2.6%) in a fertile/normozoospermic and in an obstructive azoospermic man. CONCLUSION(S): AZFc subdeletions do not seem to cause severe impairment of spermatogenesis. Moreover, gr/gr-DAZ1/DAZ2 subdeletions do not appear to affect fertility in Chilean men.