Clomiphene citrate and ovulation induction

Clomiphene can be used to treat anovulation due to hypothalamus or pituitary gland dysfunction, and it normalizes the luteal phase in stimulated patients. It can be used to estimate ovarian follicle reserve, and may be predictive of ovulation in women aged >/=35 years or with failed IVF. Contraindications include risk of congenital anomalies, chronic liver disease and visual disorders. Clomiphene may impair fertility through its effects on cervical mucus and in causing various endometrial dysfunctions. However, if clomiphene is administered in 50 mg doses, side-effects are avoided and efficacy is similar to that of a 100 mg dose, although daily dosages of 200 mg/day over 5 days can induce ovulation in approximately 70% of treated patients. Gonadotrophin concentrations increase up to days 5-9 when follicles are selected, and clomiphene is effective in patients with polycystic ovary syndrome (PCOS). Fifty percent of normal patients conceive, a value perhaps biased by the antagonistic ef

P450Arom induction in isolated control endometrial cells by peritoneal fluid from women with endometriosis

Objective: To study the effect of peritoneal fluid from women with (PF-E) and without (PF-C) endometriosis on P450Arom expression in endometrial cells. Design: Experimental study. Setting: University research unit. Patient(s): Forty women of reproductive age with (n ¼ 22) or without (control; n ¼ 18) endometriosis. Intervention(s): Peritoneal fluid and eutopic endometrial samples were obtained during surgery from women with (n ¼ 13 and 9, respectively) and without (n ¼ 4 and 14, respectively) endometriosis. Main Outcome Measure(s): Expression study for P450Arom, steroid factor 1 (SF-1), chicken ovalbumin upstream transcription factor I (COUP-TFI), and COUP-TFII messenger RNA (reverse transcriptase–polymerase chain reacion) and/or protein (immunoblot) in isolated endometrial epithelial cells transfected or not with expression vector containing SF-1, COUP-TFI, or COUP-TFII complementary DNAs. Result(s): Basal messenger RNA and/or protein expression of P450Arom and SF-1 were augmented in endometriosis, and that of COUP-TF was diminished. In control cells, (Bu)2cAMP and PF-E increased P450Arom and SF-1 expression (but not COUP-TF expression) in a dose-dependent way, an effect not observed with PF-C, adsorbed PF-E, or 10 5 M indomethacin. Transfected cells confirmed these results. Any treatments modified the studied molecules in endometriosis cells. Conclusion(s): These data indicate that molecules contained in PF-E favor an estrogenic microenvironment, suggesting a role in the etiopathogenesis of endometriosis enabling the survival, maintenance, and growth of endometrial implants in the ectopic locations

Nuclear factor κB pathway and interleukin-6 are affected in eutopic endometrium of women with endometriosis

In order to investigate the role of the nuclear factor κB (NFKB) pathway on gene expression in the eutopic endometrium in endometriosis, and in particular of interleukin-6 (IL6), we evaluated RELA, IκB kinase (CHUK), NFKBIA and IL6 expressions and NFKB DNA binding in eutopic endometrium from women with endometriosis. Eutopic endometrium was obtained from 37 women with endometriosis and 42 fertile women during laparoscopy. We analysed RELA, CHUK, NFKBIA and IL6 mRNA levels (RT-PCR); RELA, CHUK and NFKBIA proteins and p-NFKBIA/NFKBIA ratio (western blot); and NFKB binding (DNA shift assay) and IL6 concentration (ELISA) in endometrial explants. Our results indicate that mRNA and cytoplasmic proteins of RELA and CHUK exhibit constant levels in normal endometrium during the menstrual cycle. A dramatic increase (P<0.05) in NFKBIA mRNA expression, RELA nuclear presence and the mRNA and the protein of IL6 during late secretory phase was also observed in this tissue. By contrast, in eutopic en

Differential expression of upstream stimulatory factor (USF) 2 variants in eutopic endometria from women with endometriosis: estradiol regulation

BACKGROUND: Endometriosis, pro-inflammatory and invasive benign disease estrogen dependent, abnormally express in endometria the enzyme P450Arom, positively regulated by steroid factor-1 (SF-1). Our objective was to study the nuclear protein contents of upstream stimulating factor 2 (USF2a and USF2b), a positive regulator of SF-1, throughout the menstrual cycle in eutopic endometria from women with and without (control) endometriosis and the involvement of nuclear estrogen receptors (ER) and G-coupled protein estrogen receptor (GPER)-1 RESULTS: Upstream stimulating factor 2 protein contents were higher in mid (USF2b) and late (USF2a and USF2b) secretory phase in eutopic endometria from endometriosis than control (p < 0.05). In isolated control epithelial cells incubated with E2 and PGE2, to resemble the endometriosis condition, the data showed: (a) significant increase of USF2a and USF2b nuclear protein contents when treated with E2, PPT (specific agonist for ERa) or G1 (specific agonist for GPER1); (b) no increase in USF2 binding to SF-1 E-Box/DNA consensus sequence in E2-treated cells; (c) USF2 variants protein contents were not modified by PGE2; (d) SF-1 nuclear protein content was significantly higher than basal when treated with PGE2, E2 or G1, stimulation unaffected by ICI (nuclear ER antagonist); and (e) increased (p < 0.05) cytosolic protein contents of P450Arom when treated with PGE2, E2, PPT or G1 compared to basal, effect that was additive with E2 + PGE2 together. Nevertheless, in endometriosis cells, the high USF2, SF-1 and P450Arom protein contents in basal condition were unmodified CONCLUSION: These data strongly suggest that USF2 variants and P450Arom are regulated by E2 through ERa and GPER1, whereas SF-1 through GPER1, visualized by the response of the cells obtained from control endometria, being unaffected the endogenously stimulated cells from endometriosis origin. The lack of E2 stimulation on USF2/SF-1 E-Box/DNA-sequence binding and the absence of PGE2 effect on USF2 variants opposite to the strong induction that they exert on SF1 and P450 proteins suggest different mechanisms and indirect regulations. The sustained USF2 variants protein expression during the secretory phase in eutopic endometria from women with endometriosis may participate in the pathophysiology of this disease strongly associated with infertility and its characteristic endometrial invasion to ectopic sites in the pelvic cavit

CRIOPRESERVACION DE PRONUCLEOS: ROL EN EL PROGRAMA DE FERTILIZACION ASISTIDA

Objetivo: Determinar el rol de la criopreservación (CP) de pronúcleos (PN), como una herramienta para disminuir la incidencia de embarazos múltiples y dar otra oportunidad de transferencia embrionaria, sin requerir estimulación ovárica nuevamente, en parejas con infertilidad que requieren como tratamiento algún tipo de procedimiento de Fertilización Asistida (FA). Material y Método: Se analizaron los resultados de 545 procedimientos de FA entre marzo del 2000 y junio del 2003. Resultados: La incorporación de la CP se logra en diciembre del 2001, criopreservando hasta la fecha el 51,3% de las parejas que tiene más de 6 folículos a aspirar. Se han criopreservado 623 PN dando un promedio 6,4 PN por pareja. Se han descongelado 166 PN, sobreviviendo 134 PN, lo que implica un 80,7%. Ciento catorce PN clivaron a embrión de 4 células, equivalente a un 85,1%. Se han transferido 114 PN en 39 ciclos con un promedio de 2,9 embriones por pareja, dando origen a 12 embarazos clínicos (30,7%) con una tasa implantacional del 11,4%. Se observa una reducción del número de embarazos múltiples en dobles del 23% al 15,5%, en triples del 7,6% al 2,2% y la no ocurrencia de cuádruples o más, al comparar el período 2000-diciembre 2001 (sin CP de PN), con diciembre de 2001-junio del 2003<br>Program at the San Borja Arriarán Clinical Hospital - University of Chile as a method of reducing the incidence of multiple pregnancies and providing an additional opportunity of embryo transfer, without requiring another hyperstimultaion ovarian cycle. Material and Methods: Five hundred and forty five Assisted Fertilization procedures have been performed between march 2000 and june 2003. Results: The PN cryopreservation program was approved by the Institutional Review Board in december 2001. Since then 51.3% of couples participating in our IVF program have asked for PN cryopreservation. The total number of PN (n= 623) that have been cryopreserved resulting in an average of 6.4 PN per couple. Presently, 166 PN have been unfrozen with a surviving rate of 80.7% (n= 134). Eighty five percent of the PN (n= 114) cleaved to four to six cells embryo. All of the have been transferred in 39 cycles with an average of 2.9 embryos per couple, originating 12 clinical pregnancies (30.7%) with 11.4% rate of implantation. The PN cryopreservation program has diminished in a significant manner the number of multiple pregnancies in twins (23% to 15.5%), as well as, triplets (7.6% o 2.2%). In addition we have not seen quadruples or more over the time agenda of the cryopreservation progra

Ovarian function during puberty in girls with type 1 diabetes mellitus: Response to Leuprolide

Context: An increased prevalence of polycystic ovary syndrome (PCOS) has been reported in adult women with type 1 diabetes mellitus (DM1). We investigated whether these hormonal abnormalities begin during puberty by evaluating the ovarian steroidogenic response to leuprolide acetate. Methods: We studied 56 adolescent girls with DM1 (aged 12.3 +/- 0.2 yr) and 64 healthy girls (C) (aged 11.9 +/- 0.2 yr) up to 2 yr post menarche, matched by age, body mass index, and pubertal development. We evaluated anthropometrical data and Ferriman-Gallway score and performed a leuprolide test (500 mu g sc) to study ovarian function. Ovarian volume was determined by transabdominal ultrasonography. Results: We found five DM1 but no C girls with abnormally located terminal hair (Fisher's exact, P < 0.05). Free androgen index increased throughout puberty in girls with DM1 (ANOVA, P < 0.0001), which was associated with a decrease in SHBG levels in girls with DM1 (ANOVA, P < 0.0001). Stimulated 17OH progesterone (17OHProg) increased throughout puberty only in girls with DM1 (ANOVA, P < 0.01). Girls with DM1 at Tanner stage 5 had higher stimulated LH to FSH ratio, testosterone, and 17OHProg levels than girls at Tanner stage 4. In contrast, in C girls the stimulated testosterone, 17OHProg, and LH to FSH ratio were similar at Tanner stages 4 and 5. Ovarian volumes and uterine length were larger in girls with DM1 (analysis of covariance, P < 0.05). Conclusions: These data suggest that patients with DM1 have differences in ovarian steroidogenic response to leuprolide, compared with C girls during puberty. Future studies in young women should clarify whether these findings are related to the pathogenesis of hyperandrogenism later in life

CRIOPRESERVACION DE PRONUCLEOS: ROL EN EL PROGRAMA DE FERTILIZACION ASISTIDA

Objetivo: Determinar el rol de la criopreservación (CP) de pronúcleos (PN), como una herramienta para disminuir la incidencia de embarazos múltiples y dar otra oportunidad de transferencia embrionaria, sin requerir estimulación ovárica nuevamente, en parejas con infertilidad que requieren como tratamiento algún tipo de procedimiento de Fertilización Asistida (FA). Material y Método: Se analizaron los resultados de 545 procedimientos de FA entre marzo del 2000 y junio del 2003. Resultados: La incorporación de la CP se logra en diciembre del 2001, criopreservando hasta la fecha el 51,3% de las parejas que tiene más de 6 folículos a aspirar. Se han criopreservado 623 PN dando un promedio 6,4 PN por pareja. Se han descongelado 166 PN, sobreviviendo 134 PN, lo que implica un 80,7%. Ciento catorce PN clivaron a embrión de 4 células, equivalente a un 85,1%. Se han transferido 114 PN en 39 ciclos con un promedio de 2,9 embriones por pareja, dando origen a 12 embarazos clínicos (30,7%) con una tasa implantacional del 11,4%. Se observa una reducción del número de embarazos múltiples en dobles del 23% al 15,5%, en triples del 7,6% al 2,2% y la no ocurrencia de cuádruples o más, al comparar el período 2000-diciembre 2001 (sin CP de PN), con diciembre de 2001-junio del 200