RESUMO
The aim was to design a protocol combining eCG followed by hCG for estrus induction in the bitch. In Experiment 1, three ovariohysterectomized bitches received 10 000 IU of eCG iv, and 15 days later 10 000 IU of eCG im. Blood samples were taken up to 144 h after each injection to measure eCG concentrations. In Experiment 2, 25 healthy, intact late anestrous bitches were assigned to one of five doses of eCG (5, 10, 15, 20, 44, or 50 IU/kg eCG im; [TRT5-TRT50]). Sexual behavior (SB), clinical signs of estrus (CSE) and vaginal cytology (VC) samples were obtained and scored before eCG administration and every other day until onset of estrus, or for 14 days. In Experiment 3, intact late anestrous bitches were assigned to a treatment group (TRT; n = 16) and received eCG (50 IU/kg im) followed by hCG (500 IU im) 7 days later; or to a placebo group (PLA; n = 8) where they received 1 mL saline solution im. All bitches that were induced in estrus were mated or AI with fresh semen. In Experiment 1, maximum observed concentration (C(max)) eCG were similar between im and iv routes (6.1 ± 0.9 vs. 8.6 ± 0.5 IU/mL, P > 0.08), whereas time for maximum observed concentration (T(max.)) was longer for im compared to iv routes (17.5 ± 0.5 vs. 11.6 ± 0.3 h, P < 0.01). The area under the curve (AUC) was similar for im and iv routes (P > 0.48), and eCG was detectable in serum for at least 144 h for both routes. In Experiment 2, 3 days or 3 to 5 days after treatment, all bitches in TRT50 had higher scores compared to TRT5-44 animals (P < 0.01). In TRT50, the mean interval from treatment to estrus was 4.0 ± 0.4 days. In Experiment 3, the mean interval from treatment to estrus was shorter in the TRT group compared to the PLA group (4.1 ± 3.3 vs. 68.5 ± 4.4 days, P < 0.01). The previous interestrus interval was similar for TRT and PLA groups (199.6 ± 7.2 vs. 197.5 ± 10.2 days), but the new interestrus interval was shorter for the TRT compared to the PLA group (164.0 ± 7.2 vs. 212.2 ± 10.2 days; treatment by interval interaction, P < 0.007). Serum P(4) concentrations increased on the first day of cytologic diestrus after treatment in bitches in TRT (0.7 ± 0.3 vs. 22.8 ± 4.2 ng/mL; P < 0.01); but did not change in PLA (P > 0.84). Ninety-four percent of animals were bred (15/16; AI, n = 7; natural mating, n = 8), and 80% (12/15) became pregnant. None of the bitches had any side effects from the eCG and hCG therapy. We concluded that 50 IU/kg of eCG combined 7 days later with 500 IU of hCG was effective to induce normal and fertile estrus in bitches at 164 days post estrus, with an 80% pregnancy rate, with no side effects, and with a reduction of 48 days of the interestrus interval.
Assuntos
Gonadotropina Coriônica/farmacologia , Gonadotropina Coriônica/farmacocinética , Estro/efeitos dos fármacos , Animais , Área Sob a Curva , Gonadotropina Coriônica/administração & dosagem , Cães , Vias de Administração de Medicamentos , Feminino , Meia-Vida , Cavalos , Humanos , Masculino , Gravidez , Comportamento Sexual AnimalRESUMO
The purpose of this study is to provide evidence that empty follicle syndrome (EFS) is a result of an abnormality in the in-vivo biological activity of some batches of commercially available human chorionic gonadotrophin (HCG). This is a comparative study between six consecutive in-vitro fertilization (IVF) cases with EFS (study group) and 10 IVF pregnancy cycles (control group). Both groups received the same ovarian stimulation protocol consisting of leuprolide acetate and human menopausal gonadotrophin (HMG). An i.m. injection of 10,000 IU of HCG was administered once follicles had reached 18-20 mm and oestradiol/follicle > or = 16 mm was at least 900 pmol/l. Transvaginal aspiration was performed 36 h later. Plasma HCG prior to and 12 h after i.m. injection as well as the follicular fluid (FF) concentrations of oestradiol, progesterone, luteinizing hormone (LH) and HCG were determined in the study group and controls. The in-vitro biological activity of the batch of HCG used by the EFS cases and the control group was determined using a Leydig cell preparation from adult rats. Furthermore, the plasma clearance rate after i.v. injection of 5000 IU of HCG, from the same batches, was studied in three male volunteers. In the IVF cycles, no HCG was detected in plasma prior to the injection of commercial HCG. After 12 h, no HCG was detected in the study group compared to a mean of 207.5 IU/l (110-360) in controls. Mean FF concentration of LH, HCG, progesterone and oestradiol was 0.9 IU/l, 0 IU/l, 3.1 nmol/ml and 4.4 nmol/ml in EFS compared to 1.0, 98.3, 32.0 and 3.7 in pregnancy cycles. The in-vitro biological activity in both HCG batches was not significantly different; however, immunoreactive HCG used in EFS cases was undetectable in plasma of male volunteers as soon as 10 min after i.v. injection of 5000 IU of HCG. The endocrine abnormalities found in follicular fluids of EFS are not a consequence of an ovarian problem but the result of a lack of exposure to biologically active HCG. The rapid clearance of the drug after i.v. injection and the high affinity of desialylated HCG to liver cells suggest this to be a possible explanation for this infrequent but unfortunate event.
Assuntos
Gonadotropina Coriônica/farmacocinética , Fertilização in vitro , Oócitos , Folículo Ovariano/citologia , Folículo Ovariano/efeitos dos fármacos , Indução da Ovulação , Adulto , Gonadotropina Coriônica/química , Gonadotropina Coriônica/uso terapêutico , Estabilidade de Medicamentos , Estradiol/sangue , Estradiol/metabolismo , Feminino , Hormônio Foliculoestimulante/sangue , Líquido Folicular/metabolismo , Humanos , Leuprolida/uso terapêutico , Hormônio Luteinizante/sangue , Masculino , Menotropinas/uso terapêutico , Gravidez , Progesterona/sangue , Progesterona/metabolismo , SíndromeRESUMO
The scheduled administration of human chorionic gonadotropin in the ovarian stimulation schemes used in Assisted reproduction has made possible to control the ovulatory phenomenon. In order to know the biological repercussion upon the ovule as to elapsed time from HCG and the time of capture, the protocols of Gametes Laboratory, were reviewed; there were 277 cases with this factor. The capture moment was from 32.88 to 41.06 hours from HCG administration, with an average of 36.8 +/- 1.01 hours. There was no significant statistical correlation with the total amount of captured oocytes, useful oocytes, defective oocytes (atresic, broken, degenerated) segmentation rate and embryonary quality. There was a positive, lineal, statistical correlation with the amount of post-mature oocytes (r = 0.35, p = 0.019) and negative with fertilization rate (r = -0.30, p = < 0.0001), both without a correlation among themselves. It was concluded that prolonged time between HCG exposition more than difficulting oocyte capture, negatively affects oocyte quality with a diminution in its possibility of being fertilized. It must be found the optimal window that permits easily obtaining the larger amount of oocytes with the best possibilities of originating healthy pre-embryos.
Assuntos
Gonadotropina Coriônica/administração & dosagem , Fertilização in vitro/métodos , Oócitos/efeitos dos fármacos , Indução da Ovulação/métodos , Gonadotropina Coriônica/farmacocinética , Gonadotropina Coriônica/farmacologia , Feminino , Líquido Folicular , Transferência Intrafalopiana de Gameta , Humanos , Infertilidade Feminina/terapia , Oócitos/fisiologia , Gravidez , Fatores de TempoRESUMO
O presente estudo tem como objetivos comparar os parâmetros ultra-sonográficos do desenvolvimento folicular e endometrial, avaliar o potencial ovulatório de cada folículo ovariano de acordo com seu respectivo diâmetro médio (DFM) em resposta à gonadotrofina coriônica humana (HCG) e analisar a possível influência de cada um desses parâmetros nas taxas de gestaçåo de ciclos induzidos com o citrato de clomifeno (CC) e/ou gonadotrofina de mulher menopausada (HMG). Cento e três pacientes normoestrogênicas inférteis foram submetidas a 125 ciclos de hiperestimulaçåo ovariana controlada e foram divididas em dois grupos de acordo com o protocolo de estimulaçåo utilizado: grupo A (n=48) - HMG/HCG e grupo B (n=56) - CC/HMG/HCG. Verificou-se uma reduçåo de 36,4 por cento do núemro de ampolas de HMG utilizadas no grupo B (p<0,001), quando comparado ao grupo A. A distribuiçåo dos diagnósticos de infertilidade nåo revelou diferença entre os grupos. A Ultra-sonografia transvaginal seriada foi realizada diariamente, com o objetivo de avaliar o crescimento folicular, a espessura endometrial e as taxas de ovulaçåo por DFM no dia da administraçåo do HCG. As taxas de gestaçåo foram comparadas entre os grupos. As porcentagens de folículos medindo 12mm, 13-14mm, 15-16mm, 17-18mm, 19-20mm e > 20mm no dia da administraçåo do HCG, que subseqüentemente ovularam, foram 0 por cento e 0 por cento; 19,4 por cento e 23,2 por cento; 38,5 por cento e 66,1 por cento; 72,2 por cento e 91,7 por cento; 88,6 por cento e 97,6 por cento; 100 por cento e 92,3 por cento para os grupos A e B, respectivamente, nåo se observando diferença entre os dois grupos. As taxas de ovulaçåo foram significativamente menores (p<0,002, p<0,05; grupos A e B, respectivamente) para os folículos com DFM 16mm, quando comparados estes àqueles com DFM 17mm. O número total de folículos, ovulaçöes e espessura endometrial no dia da confirmaçåo da ovulaçåo nåo apresentou diferença quando ciclos com ou sem gestaçåo foram comparados. Considerando-se os ciclos com gestaçåo dos dois grupos, a distribuiçåo do DFM no dia da administraçåo do HCG, dos folículos que ovularam, foi significativamente diferente (p<0,001). A incidência de folículos com DFM 17mm no dia da administraçåo do HCG foi significativamente maior (p=0,01) nos ciclos de gestaçåo do grupo A quando comparados a estes ciclos do grupo B. Gestaçöes clínicas foram observadas em 17 (29,3 por cento) e 11 (16,4 por cento) ciclos dos grupos A e B, respectivamente, nåo se verificando diferença estatística entre os dois grupos (p=0,08). Os autores concluem que: (1) DFM 17mm no dia da administraçåo do HCG correlaciona-se com altas taxas de ovulaçåo em protocolos de hiperestimulaçåo ovariana controlada que utilizam CC e/ou HMG; (2) uma vez que nåo há diferença significativa nas taxas de gestaçåo entre os dois grupos e que o número de ampolas de HMG utilizadas em protocolo CC/HMG é menor, esta associaçåo represaenta uma opçåo atraente como primeira escolha em ciclos de hiperestimulaçåo ovariana controlada, independenetmente do diagnóstico da infertilidade; (3) o atraso na administraçåo do HCG pode ser necessário naqueles pacientes que recebem CC/HMG com a finalidade de aumentar o número de folículos com DFM 17mm e, talvez, as taxas de gestaçåo deste grupo