Reduced competence of immature and mature oocytes vitrified by Cryotop method: assessment by in vitro fertilization and parthenogenetic activation in a bovine model.

This study aimed to evaluate the embryo development competence, the nuclear maturation and the viability of germinal vesicle (GV) and metaphase II (MII) oocytes vitrified by the Cryotop method. Cumulus-oocyte complexes were derived from bovine ovaries and three experiments were conducted. In Experiment 1, GV oocytes were vitrified and underwent in vitro maturation (IVM) or not and their nuclear maturation was assessed by orcein staining. In Experiment 2, GV oocytes and MII oocytes were vitrified or not and the viability was assessed by calcein/ethidium homodimer-1 staining. In Experiment 3, MII oocytes matured before or after vitrification were submitted to in vitro fertilization (IVF) and parthenogenetic activation (PA) in order to evaluate embryo development. No difference was found for the nuclear maturation rate in the GV group (50%) and the GV control group (67%; P = 0.23) and for viability rate (56%; 77%; P = 0.055, respectively). However, in the MII group (27%) viability was significantly lower than that of the MII control group (84%; P < 0.0001). The cleavage rate by IVF and PA was similar in the GV group and the MII group. In contrast, vitrified MII oocytes showed no capacity for blastocyst development after IVF or PA and vitrified GV oocytes were able to develop to blastocysts only after PA, but not after IVF. In conclusion, oocyte vitrification by the Cryotop method reduced the capacity for embryo development. Vitrification of GV oocytes, however, did not influence the capacity of meiotic nuclear maturation and they exhibited higher viability following vitrification at the MII stage.

Leuprolide acetate reduces both in vivo and in vitro ovarian steroidogenesis in infertile women undergoing assisted reproduction.

Despite the probable inhibitory effects of GnRH analogues on ovarian steroidogenesis in vitro, their association with assisted reproduction protocols shows favorable results. This suggests that there are important differences in the behaviors of these drugs when administered in vivo versus in vitro. To clarify these differences, this study was designed to analyze the effect of leuprolide acetate (LA) on ovarian steroidogenesis in women undergoing In Vitro Fertilization (IVF). A prospective, randomized open label study was conducted on 14 women (26-35 years): seven receiving only gonadotrophins (Group 1) and seven receiving gonadotrophin plus LA at 1mg/day (Group 2). The LA in vivo effect was determined with serum and follicular fluid (FF) samples and via luteinized granulosa cell cultivation (GCC), where cells were obtained during oocyte retrieval after ovarian hyperstimulation. In vitro analysis was performed via addition of LA to GCC only for Group 1 (without LA) at progressively higher concentrations (0, 10(-12), 10(-9) and 10(-6)M). In vivo, the main observation was a reduction in androgen production in Group 2, represented by lower androstenedione production in FF (G1=6479+/-3458; G2=3021+/-1119 ng/ml; p=0.04) and a lower testosterone peak in GC at 96h (G1=0.64+/-0.12 ng/ml; G2=0.50+/-0.19 ng/ml; P=0.02), but a higher fertilization rate (G1=67%; G2=83%; p=0.009). In vitro, testosterone, estradiol and progesterone were also reduced by LA, even though this reduction occurred for progesterone only at the highest LA dosage (10(-6)M; 606.0+/-114.3 ng/ml versus 1524.0+/-246.5 ng/ml; p=0.02). Results show that LA reduces ovarian steroidogenesis in vivo by essentially inhibiting androgen synthesis; whereas, in vitro, ovarian steroidogenesis is reduced overall.

Low-dose transdermal hormone therapy does not interfere with the blood pressure of hypertensive menopausal women: a pilot study.

OBJECTIVES: To determine the effects of low-dose transdermal hormone therapy (HT) on systolic (SBP) and diastolic (DBP) blood pressure (BP) evaluated by 24-h ambulatory blood pressure monitoring (ABPM) in hypertensive postmenopausal women. METHODS: The study was conducted on 24 hypertensive postmenopausal women aged, on average, 54 years and under treatment with enalapril maleate (10-20 mg/day) combined or not with hydrochlorothiazide (25 mg/day). Thirteen women used a transdermal adhesive containing estradiol and norethisterone (25 and 125 mug active substance/day, respectively) and 11 did not receive HT. ABPM, lipid profile, and climacteric symptoms were evaluated before and 3 and 6 months after treatment. RESULTS: After 3 and 6 months of follow-up, there was a statistically significant reduction of the Blatt-Kupperman menopausal index in the treated group (19.6+/-8.3 vs. 9.6+/-5.9 vs. 9.7+/-7.0; P=0.01). No significant difference in any of the ABPM variables (areas under the systolic and diastolic curves, mean SBP and DBP, SBP and DBP loads and wakefulness-sleep variation) or in the lipid profile was observed between or within groups at the three time points studied. CONCLUSION: Low-dose transdermal HT administered for 6 months was effective in improving climacteric symptoms and did not change BP values or circadian pattern in postmenopausal women with mild-to-moderate arterial hypertension taking antihypertensive medications.

Absence of circadian salivary cortisol rhythm in women with anorexia nervosa.

STUDY OBJECTIVE: To compare the cortisol levels and 24 hour salivary cortisol rhythm in patients with anorexia nervosa (AN) and normal controls. DESIGN: Prospective transversal controlled study. SETTING: Tertiary-referral University Hospital. PARTICIPANTS: Twenty-five patients aged 15 to 35 years, 13 of them with regular ovulatory cycles, and 12 with diagnosis of AN. INTERVENTIONS: Salivary and blood collection for cortisol 24-hour rhythm determination. MAIN OUTCOME: Salivary cortisol was determined at 9 am, 5 pm, and 11 pm. Seric follicle-stimulating hormone, luteinizing hormone (LH), prolactin, estradiol (E2), progesterone, dehydroepiandrosterone-S (DHEA-S), and cortisol were sampled together with the 9 am salivary sample. RESULTS: LH, E2, and DHEA-S levels were reduced in patients with AN. A correlation between salivary and serum cortisol levels was observed in the 9 am sample only in controls (r = 0.67, P = 0.01; AN: r = 0.48, P = 0.12). Cortisol rhythm was present in all control subjects, whereas it was absent in one third of AN patients. The area under the curve for the AN group with preserved rhythm was significantly higher than for the control group (Me = 6811 ng/dl/24h vs 3708 ng/dl/24 h; P = 0.034). CONCLUSION: Patients with AN have higher salivary cortisol levels when compared to normal women and some of them do not present circadian rhythm.

Gonadal function in adolescent patients submitted to chemotherapy during childhood or during the pubertal period.

STUDY OBJECTIVE: To determine the presence of impaired gonadal function in adolescent patients submitted to chemotherapy during childhood or during the pubertal period. DESIGN: A case series study of 28 patients aged 12 to 19 years with menarche at least 2 years before the study. SETTING: Tertiary care public hospital. PARTICIPANTS: Group I: 14 adolescents previously submitted to chemotherapy during the prepubertal or peripubertal period and with remission of oncologic disease for at least 2 years; Group II: 14 normal adolescents with no previous oncologic disease and with regular menstrual cycles. INTERVENTIONS AND MAIN OUTCOME MEASURES: Pubertal development, menstrual cycles and serum levels of follicle-stimulating hormone (FSH) and luteinizing hormone (LH) were determined during the early follicular phase. RESULTS: There were no differences between the two groups in terms of age at appearance of secondary sexual characteristics or age at menarche. Menstrual irregularity was detected in 7 of the 14 patients in Group I, all 8 of whom presented oligomenorrhea. There were no differences in LH levels between the two groups (P = 0.55), although mean FSH levels were higher in Group I than in Group II (6.71 +/- 2.99 mIU/ml vs. 3.83 +/- 2.01 mIU/ml, P = 0.01). CONCLUSION: Although girls submitted to chemotherapy during the prepubertal or peripubertal period presented normal sexual development, the incidence of oligomenorrhea was higher than expected for their age, and FSH levels, although within normal limits, were higher than those seen in normally cycling girls.

[Tibolone treatment of diabetic postmenopausal women: clinical and laboratory safety parameters]. / Terapêutica com tibolona em mulheres diabéticas na pós-menopausa: parâmetros clínicos e laboratoriais de segurança.

OBJECTIVE: To determine the clinical-laboratory safety profile of tibolone treatment in postmenopausal women with NIDDM. METHOD: a prospective, longitudinal, open and uncontrolled study involving 24 postmenopausal women with NIDDM sequentially treated with placebo (6 months) and 2.5 mg/day tibolone (6 months). Clinical evaluation, anthropometric parameters, biochemical and hormonal measurements, and transvaginal ultrasonography were performed at baseline and after 6 (time 1) and 12 months of follow-up (time 2). Statistical analysis was performed by repeated measures analysis of variance, with the level of significance set at 5%. RESULTS: Side effects were present only during tibolone use (headache and mastalgia in 8.3% and genital bleeding in 16.6%). There was a significant reduction in the climacteric symptoms evaluated by Blatt-Kupperman index [22.2 +/- 7.1 (baseline) vs. 13.6 +/- 6.7 (time 1) vs. 3.1 +/- 3.3 (time 2); p< 0.0001]. After the tibolone use, we observed significant reductions in % body fat, diastolic arterial pressure, aminotransferases, triglycerides and HDL-cholesterol. There were no significant variations in systolic arterial pressure, heart rate, body mass index, waist to hip ratio, or in the glycemic, glycosylated hemoglobin, urea, total cholesterol and LDL-cholesterol levels. Ultrasonographic evaluation showed no significant changes in uterine volume or endometrial thickness. CONCLUSION: Short-term treatment with tibolone showed a good clinical-laboratory safety profile in postmenopausal women with NIDDM.

Effects of the etonogestrel-releasing contraceptive implant inserted immediately postpartum on maternal hemostasis: a randomized controlled trial.

INTRODUCTION: The puerperium is the period of highest risk for thrombosis during a woman's reproductive life and it is an important time for initiating an effective contraceptive method in order to increase intergestational interval. Thus, the objective of the present study was to evaluated the effects of the etonogestrel (ENG)-releasing contraceptive implant inserted immediately postpartum on maternal hemostasis markers during the first six weeks of delivery. MATERIALS AND METHODS: Forty healthy women aged 18 to 35 years-old were randomized to receive either the ENG-releasing implant 24-48 h after delivery (implant group; n=20) or nothing (control group) until the sixth postpartum week. Blood samples were collected at 24-48 h and at 6 weeks after delivery, and hemostatic variables, including fibrinogen, coagulation factors, protein C, free protein S, antithrombin, α2-antiplasmin, plasminogen activator inhibitor 1, thrombin-antithrombin complex (TAT), prothrombin fragment (PF)1+2, and D-dimers, as well as normalized activated protein C sensitivity ratio (nAPCsr), thrombin time, activated partial thromboplastin time, and prothrombin time were evaluated. RESULTS: Insertion of the ENG-releasing contraceptive implant did not change the physiological reduction in overall coagulation (TAT and PF1+2) and fibrinolysis (D-dimer) markers, or nAPCsr. Reductions in factors II, VII, X and fibrinogen and increases in factor V were greater in the control than in the implant group. Clotting factors remained within normal limits throughout the study. CONCLUSION: The ENG-releasing contraceptive implant inserted immediately postpartum did not have negative effects on physiological variations of the hemostatic system during the first 6 weeks postpartum.

Terapêutica com tibolona em mulheres diabéticas na pós-menopausa: parâmetros clínicos e laboratoriais de segurança / Tibolone treatment of diabetic postmenopausal women: clinical and laboratory safety parameters

OBJETIVO: determinar o perfil de segurança clínico-laboratorial da terapia com tibolona em mulheres portadoras de diabetes mellitus não-insulinodependente (DMNID). MÉTODO: estudo prospectivo, longitudinal, aberto e controlado envolvendo 24 mulheres na pós-menopausa portadoras de DMNID, tratadas seqüencialmente com placebo (6 meses) e tibolona 2,5mg/dia (6 meses). Parâmetros clínicos, antropométricos, bioquímicos, hormonais e ultra-sonográficos foram avaliados no período basal, após 6 (tempo 1) e 12 meses de acompanhamento (tempo 2). Análise estatística foi realizada utilizando-se ANOVA para medidas repetidas, com nível de significância 5 por cento. RESULTADOS: efeitos colaterais surgiram apenas durante uso da tibolona (cefaléia e mastalgia em 8,3 por cento e sangramento genital em 16,6 por cento). Houve diminuição significativa dos sintomas climatéricos avaliados através do índice de Blatt-Kuperman [22,2 ± 7,1 (basal) vs. 13,6 ± 6,7 (tempo 1) vs. 3,1 ± 3,3 (tempo 2); p< 0,0001]. Após uso da tibolona, evidenciamos reduções significativas no percentual de gordura corporal, pressão arterial diastólica, níveis de transaminases, triglicerídeos e HDL-colesterol. Não houve variações significativas na pressão arterial sistólica, freqüência cardíaca, índice de massa corporal, relação cintura/quadril, glicemia de jejum, hemoglobina glicosilada, uréia, colesterol total e LDL-colesterol. A avaliação ultra-sonográfica não revelou variações significativas do volume uterino e espessura endometrial. CONCLUSAO: o tratamento com tibolona em curto prazo mostrou bom perfil de segurança clínico-laboratorial em pacientes na pós-menopausa portadoras de DMNID.