Effect of the subdose of human chorionic gonadotropin applied in the Hou Hai acupoint on ovulation induction in mares / [Efeito da subdose de gonadotrofina coriônica humana aplicada no acuponto Hou Hai sobre a indução da ovulação em éguas]

The objective of this study was to evaluate the effects of an hCG sub dose applied at the Hou Hai acupoint on corpus luteum (CL) quality and ovulation induction in mares. Fifteen crossbred mares were distributed in randomized blocks and used in three periods with each period employed as the blocking factor in three treatments: T1 = 1500 IU of hCG via intravenous (IV); T2 = 450 IU of hCG applied at the false acupoint (IV); and T3 = 450 IU of hCG applied at the Hou Hai acupoint. Mean diameter of the CL, serum concentration of progesterone (P4), vascularization of the pre-ovulatory follicle and CL were evaluated. Females administered 450 IU of hCG at the Hou Hai acupoint exhibited greater ovulation rates (33.33%) 48h after induction; The minimum number of colored pixel (NCP) of the pre-ovulatory follicle of control females was superior (40.33) to that of mares administered 450 IU of hCG IV at the false acupoint (36.84) and similar to that of those administered hCG at the Hou Hai acupoint (39.31). Further, moderately positive correlations were found between the CL diameter and the P4 concentration on D8 (P<0.05). IV administration of 450 IU of hCG or at the Hou Hai acupoint was efficient at inducing ovulation and ensuring the quality of CL in mares.(AU)

O objetivo foi avaliar os efeitos de uma subdose de hCG aplicada no acuponto Hou Hai na qualidade do corpo lúteo (CL) e na indução da ovulação em éguas. Quinze éguas mestiças foram distribuídas em blocos ao acaso, sendo o período utilizado como fator de blocagem, em: T1 = 1500 UI de hCG por via intravenosa (IV); T2 = 450 UI de hCG aplicado no falso acuponto (IV) e T3 = 450 UI de hCG aplicada no acuponto Hou Hai. Avaliou-se diâmetro médio do CL, concentração sérica de progesterona (P4), vascularização do folículo pré-ovulatório e do CL. As fêmeas que receberam 450 UI de hCG no acuponto Hou Hai apresentaram maiores taxas de ovulação (33,33%) 48h após a indução. O número de pixels coloridos (NPC) mínimo do folículo pré-ovulatório das fêmeas do grupo controle foi superior (40,33) ao das éguas que receberam 450 UI de hCG IV no falso acuponto (36,84) e semelhante ao das éguas que receberam hCG no acuponto Hou Hai (39,31); correlações moderadamente positivas foram encontradas entre o diâmetro do CL e a concentração de P4, ambos no D8 (P <0,05). A administração IV de 450 UI de hCG ou no acuponto Hou Hai foi eficiente na indução da ovulação e na garantia da qualidade do CL nas éguas.(AU)

Ultrassonografia testicular no modo color Doppler∕ espectral e avaliação andrológica de garanhões tratados com gonadotrofina coriônica humana (hCG) em diferentes estações do ano / [Testicular ultrasonography in the doppler / spectral color mode and andrological evaluation of stallions treated with human corionic gonadotrofin (hCG) in different seasons of the year]

Utilizaram-se quatro garanhões nos meses de janeiro, abril, julho e outubro de 2016, em dois protocolos, em que: GI (n=4; 5mL de solução salina, i.v.) e GII (n=4; 5000UI de hCG, i.v.) e subdividiram-se esses protocolos em ciclos (C1 e C2), seguindo o esquema crossover, sendo: CI=animais 1 (GI) e 2 (GII), avaliados nos dias D1, D3 e D5, e animais 3 (GI) e 4 (GII), em D2, D4 e D6; CII= animais 1 (GII) e 2 (GI), em D1, D3, D5, e animais 3 (GII) e 4 (GI), em D2, D4 e D6. Realizou-se o tratamento em D1 e D2 de cada ciclo e a ultrassonografia testicular no modo color Doppler e Doppler espectral, uma hora antes de cada coleta de sêmen e imediatamente após. Avaliou-se: número de reflexo de Flehmen, de montas sem ereção, início da monta, tempo de reação à ereção e total da monta e análises de qualidade seminal. Estatisticamente foram utilizados os testes de qui-quadrado e ANOVA. Não houve diferenças estatísticas (P>0,05) entre os parâmetros analisados. Concluiu-se que uma única dose de 5000UI de hCG em garanhões não causou alterações significativas nos parâmetros avaliados em diferentes estações do ano.(AU)

Four stallions were used in January, April, July and October 2016 in two protocols: GI (n=4; 5ml saline, iv) and GII (n=4; 5000 hCG, iv), and these protocols were subdivided into cycles (C1 and C2) following the cross over scheme, as follows: CI=animal 1 (GI) and 2 (GII) evaluated on days D1, D3 and D5 and animal 3 (GI) and 4 (GII) at D 2, D 4 and D 6; CII=animal 1 (GII) and 2 (GI) at D1, D3, D5 and animal 3 (GII) and 4 (GI) at D2, D4 and D6. Treatment was performed on D1 and D2 of each cycle and testicular ultrasound in color Doppler and spectral Doppler mode, one hour before each semen collection and immediately after. We evaluated: Flehmen's reflex number, mounts without erection, start of the mount, reaction time to erection and total mount and seminal quality analyzes. Statistically, the Chi-square and ANOVA tests were used. There were no statistical differences (P>0.05) between the analyzed parameters. It was concluded that a single dose of 5000IU hCG in stallions did not cause significant changes in the parameters evaluated in different seasons of the year.(AU)

Progesterone level on the day of hCG administration in relation to the pregnancy rates of patients undergoing assisted reproduction techniques / Valor da progesterona no dia do hCG em relação à taxa de gravidez de pacientes submetidas às técnicas de reprodução assistida

ABSTRACT Objective To evaluate the predictive capacity for pregnancy of the progesterone level on the day of administering human chorionic gonadotropin, in women submitted to assisted reproductive techniques. Methods An observational study with 914 women submitted to assisted reproductive techniques from August 2014 to June 2016. Results Total pregnancy rate was 34.58%; in that, the pregnancy rate in women <35 years, between 35 and 38, and >38 years was, respectively, 42.3%, 38.7% and 16.1% (p<0.001). For embryo transfer in the same cycle, and progesterone of 1.3ng/dL, sensitivity was 4.78%, specificity, 84.18%, accuracy, 56.72%, positive likelihood ratio of 0.3019, and negative likelihood ratio of 1.1312, with receiver operating characteristic curve of 0.46 (95%CI: 0.42-0.49). Conclusion The progesterone level on the day of administering human chorionic gonadotropin of 1.3ng/dL differs from that empirically adopted at the study site (1.7ng/dL), and has a better predictive capacity for pregnancy in the patients studied. However, the low sensitivity of this examination raises questions about its real importance.

RESUMO Objetivo Avaliar a capacidade preditiva para gravidez do valor de progesterona no dia da administração da gonadotrofina coriônica humana em mulheres submetidas às técnicas de reprodução assistida. Métodos Estudo observacional com 914 mulheres submetidas a reprodução assistida de agosto de 2014 até junho de 2016. Resultados Engravidaram 34,58% das mulheres, sendo a taxa de gravidez naquelas >35 anos, entre 35 e 38 anos e >38 anos, respectivamente, de 42,3%, 38,7% e 16,1% (p<0,001). Para o valor de progesterona até 1,3ng/dL visando à transferência embrionária no mesmo ciclo, encontraram-se sensibilidade de 4,78%, especificidade de 84,18%, acurácia de 56,72%, razão de verossimilhança positiva de 0,3019 e razão de verossimilhança negativa de 1,1312, com área sob a curva característica de operação do receptor de 0,46 (IC95%: 0,42-0,49). Conclusão O valor de progesterona no dia da administração de gonadotrofina coriônica humana até 1,3ng/dL difere daquele empiricamente adotado no local do estudo (1,7ng/dL) e apresenta melhor capacidade preditiva para gravidez nas pacientes estudadas. No entanto, a baixa sensibilidade deste exame abre questionamentos sobre sua real importância.

Effects of human chorionic gonadotropin on the normal testicular tissue of rats

PURPOSE: To investigate the effects of human chorionic gonadotropin (hCG) on the testicular tissue of young male rats. METHODS: Male Wistar rats were assigned to groups (10 rats/group).Control Group received subcutaneous saline solution; Group 1 received hCG 50UI/Kg/dose; and Group 2 received hCG 100UI/Kg/dose, daily for 15 days. Half was submitted to bilateral orchiectomy on the 16th day and the other half 45 days after the beginning of the hormone application. Testicles were weighed, measured and has their volumes determined. The diameter of the tubules and the thickness of the seminiferous epithelium were measured. RESULTS: Control Group presented the highest values of testicles volume and weight. Rats in the Control presented normal histology. In G1 and G2 atrophy of the seminiferous tubules, apoptosis of germ cells and multinucleated giant cells were observed. Comparing groups, in the first operation Control rats had higher diameter values. In the second operation, the Control was only different from G1. As for thickness, Control had higher values in both operations. Comparing the time of operation, the diameter values were higher in G1 and G2 in the second operation. For all groups, the thickness of the epithelium was higher in the second operation. CONCLUSIONS: Human chorionic gonadotropin is gonadotoxic in rats. This effect was temporary and can affect reproductive potential. The total recovery of testicular damage in the studied range could not be proved, and the effects were not dose-dependent. .

Assessment of serum progesterone level on the day of hCG injection in infertile polycystic ovarian syndrome patients referred to women's hospital, Tehran, 2009

Background: Polycystic ovarian syndrome [PCOS] is one of the most common causes of endocrine disorders and main reasons for infertility due to unovulation and recurrent abortions. There is no consensus on effect of serum progesterone level on the day of human chorionic gonadotropin [hCG] injection. This study aims to evaluate the effect of plasma levels of progesterone on the day of hCG injection on the rate of pregnancy in in vitro fertilization [IVF] cycles of PCOS cases

Materials and Methods: A stratified cohort study was conducted over a period of one year [2009] on 38 infertile women with PCOS who were suitable candidates for the IVF program. Patients were evaluated for other causes of infertility with hysterosalpingography [HSG], laparoscopy and normal sperm analysis. Patients were placed on the long protocol, followed by oocyte pick up, and finally IVF-embryo transfer [ET]. Study patients were grouped according to progesterone levels of greater or less than 1.2 ng/ml on the day of hCG injection. Pregnancy rates were defined in each group. Levels on day of hCG day clinical pregnancy outcome were assessed. Experimental data were then compared against Fisher's exact test in SPSS version 18

Results: The overall pregnancy rate in this study was 26.3%. In the group with progesterone levels more than 1.2 ng/ml on the day of hCG injection, the clinical pregnancy rate was 4 [21.1%] and chemical pregnancy rate was 3[15.8%]. In the group with progesterone levels less than 1.2 ng/ml, the clinical pregnancy rate was 1[5.3%] and chemical pregnancy rate was 2[10. 5%]

Conclusion: This study showed that PCOS patients with progesterone levels more than 1.2 ng/ml on the day of hCG injection resulted in higher chemical and clinical pregnancy rates. However, no significant statistical differences were found between the two groups. For further verification, we recommend additional studies with larger numbers of subjects

Association between serum estradiol level on the hCG administration day and IVF-ICSI outcome

Estradiol [E[2]] is required for follicular development and lay an important role in embryo implantation. The aim of this study was to assess the impact of serum E[2] levels on the day of hCG administration in IVF-ICSI patients who are performed controlled ovarian hyperstimulation [COH]. A total of 203 women who were undergone one time IVF cyclus were evaluated in this cross sectional study. All the patients were treated either with long protocol or with microdose flare protocol. The patients were categorized into five groups according to the serum E[2] levels on the day of hCG administration. The mean number of the retrieved oocytes was [NRO] 10.6 +/- 6.7, mean fertilization rate was 55.7 +/- 24.8, and implantation rate was 9.0 +/- 19.2. Of 203 patients, 43 [21%] patients were pregnant. When the overall results are examined, the number of the retrieved oocytes and the number of transferred embryos were better in patients with serum E[2] levels >4000 pg/ml and these values were statistically significant. There were no statistical difference in patients 37 years or older. In women 4000 pg/ml. In site of the lack of high quality evidence to support a positive association between serum E[2] levels and IVF-ICSI outcomes, this study shows that high E[2] levels during COH might be associated with an increased potential of pregnancy depending on better ovarian response. When the overall results are examined, the best scores were in patients with serum E[2] levels >4000 pg/ml

Ovarian Hyper-Response to Administration of an GnRH-Agonist Without Gonadotropins

Several case reports have indicated that a small subgroup of patients may develop ovarian hyperstimulation following the administration of gonadotropin-releasing hormone agonists (GnRHa) without gonadotropins. However, since only few such cases have been published, it is unclear what course to follow in subsequent cycles after ovarian hyperstimulation in the first cycle using only GnRHa. A 33-yr-old woman was referred to in vitro fertilization for oocyte donation. A depot preparation (3.75 mg) of tryptorelin without gonadotropins induced ovarian multifollicular enlargement with high estradiol level, and was followed by human chorionic gonadotropin administration and oocyte retrieval. In a subsequent cycle of the same patient, a low dose of tryptorelin (0.05 mg) did not induce ovarian hyperstimulation, and resulted in clinical pregnancy. This report shows potential management of ovarian hyperstimulation following the administration of GnRHa without gonadotropins.

Alternativas de prevenção da síndrome de hiperestimulação ovariana / Ovarian hyperstimulation syndrome prevention alternatives

A Síndrome de Hiperestimulação Ovariana (SHEO) é uma complicação iatrogênica e potencialmente fatal, resultante da estimulação ovariana excessiva em resposta às gonadotrofinas exógenas. As principais características desta síndrome são: aumento da permeabilidade capilar que leva a uma grande troca de fluído do espaço intravascular para o terceiro espaço; e crescimento excessivo do volume ovariano. Várias estratégias para a prevenção desta síndrome foram propostas: infusão de albumina, coasting, anti-inflamatórios, bloqueio do sistema renina-angiotensina, agonistas dopaminérgicos, redução da dose de gonadotrofina coriônica humana (hCG), substituição da hCG e maturação in vitro de oócitos. Nesta revisão, as evidências atuais que suportam tais estratégias, de maneira a permitir que o médico escolha a melhor alternativa, foram avaliadas

Ovarian Hyperstimulation Syndrome (OHSS) is an iatrogenic and potentially life-threatening complication, which results from an excessive ovarian stimulation to exogenous gonadotropins. The main characteristics of this syndrome are: increased capillary permeability with a massive shift of fluid from the intra-vascular compartment to the third space; and an excessive increase of ovarian volumes. Several strategies for the prevention of this syndrome were proposed: albumin infusion, coasting, anti-inflammatory drugs, renin-angiotensin system blockage, dopaminergic agonists, human chorionic gonadotrophin (hCG) dose reduction, hCG substitution and in vitro maturation of oocytes. In this review, the current evidences to support these strategies were evaluated, in order to permit that the physician chooses the best alternative

Cessation of Gonadotropin-Releasing Hormone Antagonist on Triggering Day: An Alternative Method for Flexible Multiple-Dose Protocol

This study was performed to analyze retrospectively outcomes of stimulated in vitro fertilization (IVF) cycles where the gonadotropin-releasing hormone (GnRH) antagonist was omitted on ovulation triggering day. A total of 92 consecutive IVF cycles were included in 65 women who are undergoing ovarian stimulation with recombinant FSH. A GnRH antagonist, cetrorelix 0.25 mg/day, was started when leading follicle reached 14 mm in diameter until the day of hCG administration (Group A, 66 cycles) or until the day before hCG administration (Group B, 26 cycles). The duration of ovarian stimulation, total dose of gonadotropins, serum estradiol levels on hCG administration day, and the number of oocytes retrieved were not significantly different between the two groups. The total dose of GnRH antagonist was significantly lower in Group B compared to Group A (2.7+/-0.8 vs. 3.2+/-0.9 ampoules). There was no premature luteinization in the subjects. The proportion of mature oocytes (71.4% vs. 61.7%) and fertilization rate of mature (86.3+/-19.7% vs. 71.8+/-31.7%) was significantly higher in Group B. There were no significant differences in embryo quality and clinical pregnancy rates. Our results suggest that cessation of the GnRH antagonist on the day of hCG administration during a flexible multiple-dose protocol could reduce the total dose of GnRH antagonist without compromising IVF results.

A administração de hCG incrementa a incidência de ovulações duplas e múltiplas em éguas da raça Campolina apresentando folículos codominantes? / Does hCG administration increase the incidence of double and multiple ovulations in Campolina mares with co-dominant follicles?

A gonadotrofina coriônica humana (hCG) é rotineiramente empregada na indução de ovulação em éguas. A incidência de duplas ovulações é variável, sendo comum em determinadas raças brasileiras, como Brasileiro de Hipismo e Campolina. O objetivo deste trabalho foi avaliar se a administração de hCG em éguas da raça Campolina incrementa a incidência de ovulações duplas e múltiplas. Através de palpação retal e ultrassonografia, foram avaliados dois ciclos estrais de 104 éguas, divididas em dois grupos. À detecção de um ou mais folículos ³35 mm e edema endometrial grau 2, 2.500 IU de hCG (IV) eram administrados às éguas do grupo I. O grupo II serviu como controle. Os resultados obtidos foram analisados pelo Qui-quadrado. A incidência de ovulações duplas nos grupos I e II foi de, respectivamente, 32,7% (34/104) versus 30,8% (32/ 104). Ambos os grupos obtiveram incidência semelhante de ovulações triplas (1,9% - 2/104). Apenas uma ovulação quádrupla (1,0% - 1/104) foi detectada, no grupo controle. A incidência de duplas e múltiplas ovulações foi similar entre os grupos (p > 0,05). Pelo fato das éguas da raça Campolina apresentarem, fisiologicamente, considerável incidência de ovulações duplas e múltiplas, é possível que a administração de hCG não tenha influenciado estes padrões ovulatórios.

The human chorionic gonadotrophin (hCG) has been routinely used in the equine species to induce ovulations for more than four decades. The incidence of multiple ovulations in horses is variable, being double ovulations a common occurrence in some Brazilian breeds, such as Brasileiro de Hipismo and Campolina. The aim of this work was to evaluate the role of hCG administration to increase, in Campolina mares, the incidence of double and multiple ovulations, which are desirable in embryo transfer programs. One hundred and four (104) mares had their estrous cycles recorded through rectal palpation and ultrasonographic evaluation during the breeding season of 2003 - 2004, in a total of two hundred and eight (208) cycles (two cycles per mare). Animals were allocated in two groups, with fifty two (52) horses each. In Group I mares, once one or more = 35 mm follicles and grade 2 uterine edema were present, 2.500 IU of hCG were intravenously administered. The untreated mares in Group II served as control. The Chi-Square test was used to analyze obtained results. The incidence of double ovulations in groups I and II was, respectively, of 32.7% (34/104) versus 30.8% (32/104). The incidence of triple ovulations was low in both groups, of 1.9% (2/104). Only once in the control group (1.0% - 1/104) was a quadruple ovulation detected. There was no difference in the incidence of double and multiple ovulations between both groups (p > 0.05). Possibly, hCG administration did not influence ovulation patterns in this work due to the fact that Campolina breed mares physiologically show a considerable incidence of double and multiple ovulations.

Conception rate in Holstein cows treated with GnRH or hCG on the fifth day post artificial insemination during summer / Taxa de concepção de vacas Holandesas tratadas com GnRH ou hCG no quinto dia após a inseminação artificial no verão

Lactating Holstein cows (n=158), at 213±112 days in milking and averaging 26±9kg of milk per day, were randomly assigned to one of three treatment groups: control (CG, n=52, saline), GnRH (GG, n=55, 100g gonadorelin), and hCG (HG, n=51, 2500IU) given five days after artificial insemination (AI). Rectal temperature was taken at the moment of AI and blood samples were collected five, seven, and 12 days after AI. Pregnancy was determined between 42 and 49 days after AI. Concentration of progesterone (P4) in serum (ng/ml, mean±SE) for CG, GG, and HG were, respectively, 2.7±0.4, 2.5±0.4, and 3.2±0.5 on day 5; 4.8±0.4, 4.2±0.4, and 5.7±0.5 on day 7; and 5.2±0.4, 6.9±0.4, and 8.5±0.5 on day 12 after AI. P4 concentration had proportional increase in serum between days 5 and 7 after AI (CG: 178 percent, GG: 168 percent, and HG: 178 percent), suggesting that the treatments did not induce a luteotropic effect on the existing corpus luteum (CL). Concentrations of P4 increased between days 7 and 12 in cows treated with GnRH and hCG (GG: 164 percent, and HG: 149 percent, P<0.01); but not in control cows (GC: 18 percent, P=0.31), suggesting that a new CL was formed. Treatments with GnRH or hCG increased conception rates in cows with rectal temperature below 39.7ºC (CG: 10.1 percent, n=26, GG: 36.8 percent, n=27; and HG: 32.8 percent, n=21), but not in cows with rectal temperature above 39.7ºC (CG: 15.2 percent, n=26; GG: 17.8 percent, n=28; and HG: 24.4 percent, n=30). These data suggest that high body temperature masked the positive impact of treatment with GnRH or hCG on day 5 after AI on conception.

Vacas da raça Holandesas em lactação (n=158) aos 213±112 dias de lactação e produção de 26±9kg leite/dia, foram aleatoriamente distribuídas em três grupos: controle (GC, n=52, salina); GnRH (GG, n=55, 100mcg de gonadorelina); e hCG (GH, n=51, 2500UI de hCG) aplicado no dia 5 após a inseminação artificial (IA). A temperatura retal foi verificada no momento da IA, e as amostras de sangue coletadas nos dias 5, 7 e 12 após a IA. A concepção foi determinada entre os dias 42 e 49 após IA. As concentrações séricas de progesterona (P4 - ng/ml, média±EPM) para GC, GG, e GH foram, respectivamente: no dia 5: 2,7±0,4, 2,5±0,4 e 3,2±0,4; no dia 7: 4,8±0,4, 4,2±0,4 e 5,7±0,5; e no dia 12 após a IA: 5,2±0,4, 6,9±0,4 e 8,5±0,5. O aumento proporcional na concentração sérica de P4 entre os dias 5 e 7 após IA (GC: 178 por cento, GG: 168 por cento, e GH: 178 por cento) sugere que os tratamentos não induziram efeito luteotrópico no corpo lúteo (CL) existente. O aumento na P4 sérica entre os dias 7 e 12 nos animais tratados com GnRH ou hCG (GG: 164 por cento e GH: 149 por cento, P<0,01) em relação aos animais controle (GC: 18 por cento, P=0,31), sugere a indução de novo CL. Os tratamentos com GnRH ou hCG aumentaram as taxas de concepção nas vacas com temperatura retal abaixo de 39,7ºC (GC: 10,1 por cento, n=26; GG: 36,8 por cento, n=27 e GH: 32,8 por cento, n=21), mas não em vacas com temperatura retal acima de 39,7ºC (15,2 por cento n=26; 17,8 por cento, n=28 e 24,4 por cento, n=30). Os resultados sugerem que a alta temperatura corporal pode mascarar os efeitos positivos do tratamento com GnRH ou hCG no dia 5 após a IA, na concepção.

[ effect of HCG on in vitro maturation of human immature oocytes and rate of 8 cell embryos mediated by ICSI]

The infertility treatment by using in vitro matured oocyte has many potential applications. It minimizes the risk of ovarian hyper stimulation and is an alternative treatment for women with polycystic ovarian syndrome who may have problems regarding stimulation for IVF. This method may prove important for subjects in need of fertility preservation and also provides information about the final stage of oocyte maturation. The aim of this study was to determine the effect of HCG on the rate of in vitro matured oocyte and 8 cell embryos mediated by ICSI. This experimental study was done in Montaserieh IVF center and Islamic Azad University at Mashhad. One hundred sixty eight immature oocytes aspirated from women undergoing IVF cycles were divided into 3 equal groups and allocated for maturation in 3 medium cultures. All of 3 culture groups contained G1 medium supplemented with HSA 10% [Human Serum Albumin]. Control group had no gonadotropin, but the first experimental group contained HCG 10 ILJ/ml and the second experimental group contained HCG 5 ILJ/ml. Matured oocytes were fertilized by ICSI and numbers of 8-cell embryos were investigated 72 hours after fertilization by invert microscope. The results showed that both of the HCG concentrations significantly increased the rate of matured oocytes and also the rate of 8-cell embryos with respect to control group [p<0.05]. No significant differences were found in rate of matured oocytes between the two experimental groups [p>0.05] while the number of 8-cell embryos was significantly higher in HCG 5IU/ml with respect to HCG 10 IU/ml and control group [p<0.05]. The results suggests that both of the HCG concentrations increase the rate of maturation of immature oocytes and the rate of 8-cell embryos but HCG [5IU/ml] has more effect on the number of 8-cell embryos mediated by ICSI

Indução de estro em cabras da raça Toggenburg com dois diferentes dispositivos intravaginais / Estrous induction in Toggenburg goats using intravaginal devices

Doze cabras nulíparas da raça Toggenburg foram distribuídas em dois tratamentos (T1 e T2) para indução do estro. O T1 (n=6) consistiu na inserção da esponja impregnada com 60mg de acetato de medroxiprogesterona, associado à aplicação de 50æg do análogo sintético d-cloprostenol (PGF). No quarto dia, foram aplicadas 250UI de gonadotropina coriônica eqüina (eCG) e, no quinto, a esponja foi retirada. O T2 (n=6) diferiu somente quanto ao dispositivo empregado, que foi o CIDR-G®. Seis cabras do T1 e seis do T2 entraram em estro depois da retirada dos dispositivos, sendo que 83,3 (5/6) e 33,3 por cento (2/6) ficaram gestantes em T1 e T2, respectivamente. O intervalo da retirada do dispositivo ao início do estro não diferiu entre tratamentos (P>0,05). As características ovulatórias não foram influenciadas pelos tratamentos. O intervalo da retirada do dispositivo à ovulação, para ambos os protocolos de indução do estro, não diferiu (P>0,05) entre tratamentos. Ambos os dispositivos foram eficientes em induzir o estro.

Twelve Toggenburg breed nuliparous female goats were randomly assigned to two treatments intended to induce estrus. On day zero, animals on T1 (n=6) were treated with an intravaginal sponge impregnated with 60mg of medroxyprogesterone acetate plus 50æg of PGF analogue. The T1 goats received eCG (250IU) on day 4, and on day 5 the sponge was removed. The T2 goats (n=6) received CIDR-G® as intravaginal sponge plus 50mg of PGF analogue. All goats expressed behavioral estrus, and the pregnancy rates were 83.3 (5/6) and 33.3 percent (2/6) for T1 and T2, respectively. The interval from withdrawal of the device to onset of estrus did not differ between treatments (P>0.05). Ovulatory traits also were not affected by treatments. The interval between CIDR-G® removal and ovulation for T1 and T2 goats were similar (P>0.05). Both sponge and CIDR-G were equally efficient in inducing the estrus behavior in goats.

Taxa de gestação em cabras Alpinas e Saanen tratadas com hCG no terceiro dia após o estro / Pregnancy rate and progesterone concentration of Alpine and Saanen goats treated with hCG the third day after estrus

Avaliou-se o efeito da aplicação da gonadotrofina coriônica humana (hCG) sobre a taxa e duração da gestação e sobre a concentração plasmática de progesterona (P4) em cabras lactantes das raças Alpina (83) e Saanen (60), na estação de acasalamento induzida pelo programa de fotoperíodo artificial. Os animais foram distribuídos em dois tratamentos após a cobrição; as cabras do tratamento 1 (T1 controle) receberam solução salina via intramuscular (1ml), e as do tratamento 2 (T2) receberam 250UI do hCG, no terceiro dia no pós-estro. As taxas de gestação, detectadas por exame ultra-sonográfico, realizado no 35° após a cobrição, para as cabras das raças Alpina e Saanen controle e tratadas, foram de 80,4; 70,3; 72,7; 81,5 por cento, respectivamente, não diferindo entre os tratamentos e entre as raças (P>0,05). A taxa média de gestação foi 77,2 e 75,0 por cento para os animais controle e tratados, respectivamente. A duração da gestação não foi influenciada pelos tratamentos (P>0,05). As amostras de sangue para análise da concentração plasmática de progesterona (P4) foram coletadas de cinco cabras Alpina por tratamento, no dia do estro (dia 0) e aos 3, 8, 15, 21, 42 e 60 dias após o estro. Não houve diferença na concentração plasmática média de P4 entre os tratamentos (P>0,05), sendo de 5,84ng/ml para T1 e 5,76ng/ml para T2. Observou-se diferença em relação aos dias de coleta (P<0,05). O hCG aplicado no terceiro dia pós-estro não alterou a taxa de gestação e nem a concentração de P4.

Indução da ovulação em vacas com gonadotrofina coriônica humana (hCG) purificada por cromatografia de afinidade / Induction of ovulation in cows with affinity chromatography-purified Human Chorionic Gonadotropin (hCG)

O objetivo do presente estudo foi purificar o hCG contido em uma preparação comercial (Pregnyl® Organon) por cromatografia de afinidade e avaliar a taxa de ovulação de vacas injetadas com o hormônio purificado. Uma coluna de cromatografia contendo matriz de Concanavalina-A Sepharose (Con-A) foi equilibrada e cem milunidades internacionais (UI) de Pregnyl® foram aplicadas na coluna. Frações de 3,8mL foram colhidas (4 º C) a cada 5 minutos durante 12,5 horas, resultando em um total de 150 frações. Após a fração 76 adicionou-se à coluna solução tampão contendo 0,3M de alfa−metilglicosidase. A concentração protéica das frações foi estimada por espectrofotometria (280nm). Foram observados dois picos de absorbância, um entre as frações 14 e 19 e outro entre as frações 90 e100. As frações 14 a 19 e 90 a 150 foram reunidas seqüencialmente aos pares, concentradas por ultrafiltração e analisadas por SDS-PAGE. O primeiro pico resultou da eluição das proteínas não adsorvidas pela matriz de Con-A, enquanto o segundo representou as proteínas adsorvidas pela coluna, especialmente o hCG. As frações contendo hCG foram reunidas em uma única amostra cuja concentração protéica foi quantificada pelo método de Lowry. Vacas no 5º dia de um ciclo estral sincronizado, apresentando um folículo dominante maior que 8mm de diâmetro, foram injetadas com 0,1, 0,2 ou 0,3mg de proteína contida na amostra purificada. Após 48 horas, a taxa de ovulação observada foi de 0%, 50% e 75%, respectivamente. Foi demonstrado no presente estudo que a técnica de cromatografia por afinidade foi eficiente para aumentar a concentração relativa do hCG preservando sua atividade biológica.

Objective of the present study was to purify hCG contained in commercial preparation (Pregnyl®- Organon) by affinity chromatography and to evaluate ovulation rates in cows injected with the purified hormone. A chromatography column containing concanavalin-A sepharose (Con-A) matrix was equilibrated and one hundred thousand international units (UI) of Pregnyl® were applied to the column. Fractions of 3,8mL were collected (4 degree C) every 5 minutes for 12,5h, to yield a total of 150 fractions. After fraction 76, a column buffer containing 0,3M alpha-methylglicoside was added to the column. Protein concentrations were estimated by espectrophotometry(280nm). Two peaks of absorbance were observed, between fractions 14 and 19, and fractions 90 and 100. Fractions 14-19 and 90-150 were sequentially grouped in pairs concentrated by ultrafiltration andanalyzed by SDS-PAGE. The first peak of absorbance resulted from proteins contained in the commercial product, eluted from the column and not adsorbed by the Con-A matrix. The second peak represented proteins adsorbed by the column, especially hCG. The fractions containing hCG were grouped in a single sample and protein concentration was measured by the Lowry method. Cows in day 5 of the estral cycle, which had a dominant follicle larger than 8mm, were injected with 0,1, 0,2 or 0,3mg protein of the purificated sample. The ovulation rate 48 hours after treatment was 0%, 50% and 75%,respectively. It was demonstrated that relative concentration of hCG can be increased by affinity chromatography and biological activity of purified hCG was satisfactory preserved.

Uso do hCG, GnRH ou 17beta-Estradiol e progesterona associados ao acetato de melengestrol e prostaglandina f2alfa em novilhas cruzadas (Bos taurus indicus x Bos taurus taurus) / Use of hCG, GnRH or 17beta-estradiol and progesterone associated with melengestrol acetate and prostaglandin F2alfa in cross-bred heifers (Bos taurus indicus x Bos taurus taurus)

O objetivo do presente experimento foi avaliar a performance reprodutiva e as características de estros em novilhas cruzadas de corte (Bos taurus indicus x Bos taurus taurus) tratadas com o protocolo MGA/Prostaglandina (PG) em associação a outros hormônios. No dia 0 (Dia 0 = início da ingestão do MGA) as novilhas foram distribuídas ao acaso para receber 2mL de solução salina (grupo Salina), 2500UI de hCG (grupo hCG), 20müg de acetato de buserelina (grupo GnRH) ou 5mg de 17beta-estradiol + 100mg de progesterona (grupo 17beta-E2 +P4. Amostras de sangue foram colhidas nos dias - 7, 0, 7 e 10 para a mensuração das concentrações plasmáticas de progesterona. Independentemente do tratamento, todas as novilhas receberam 0,5mg MGA/ animal/ dia durante 8 dias (Dia 0 ao 7) e PG (Dia 7). Os estros foram observados durante 120h, a partir da injeção de PG, pelo sistema Heat- Watch. As novilhas foram inseminadas 12 horas após o início dos estros ou 72h depois da injeção de PG. O diagnóstico de gestação foi realizado 35 dias após a última inseminação, por ultrasonografia. As respostas em estros foram de 50,0%, 22,2%, 59,5% e 71,8% para os grupos Salina, hCG, GnRH e 17beta-E2 +P4, respectivamente (P< ou =0,01). O intervalo médio da injeção de PG ao estro foi de 72,8 + ou - 22,2, 102,0 + ou - 22,7,84,6 + ou - 19,0 e 72,5 + ou - 24,4 horas (P< ou =0,01), sendo o grau de sincronização similar entre os grupos. As taxas de concepção foram de 57,9%, 37,5%, 40,9% e 39,3% e as taxas de prenhez de 29,0%, 11,1 %,27,0% e 28,2%, para os grupos Salina, hCG, GnRH e 17beta-E2 +P4, respectivamente. Não houve efeito do tratamento na duração dos estros (10,4 + ou - 5,7 horas), número de montas (23,0 + ou - 16,9) e duração das montas (2,7 + ou - 0,3 segundos), sendo os valores entre parênteses correspondentes às médias gerais e desvios padrão. Em conclusão, a performance reprodutiva não foi alterada pela adição do hCG, GnRH ou 17beta-E2 +P4 ao protocolo de sincronização MGA/PG.

Ovarian monosaccharides (glucose and fructose): hormonal effects and their role in final oocyte maturation and egg quality in catfish Heteropneustes fossilis, Bloch.

In the catfish H. fossilis, ovarian glucose and fructose levels showed significant annual variations that could be correlated with the gonado-somatic index. The concentration of the sugars, which was low in resting phase of gonad increased with the onset of oogenesis, reaching the peak in prespawning (June) phase. Glucose is the predominant monosaccharide in the late oogenesis, while fructose in early oogenesis and resting phase. Administration of a single injection of HCG (100 IU/fish) decreased glucose at 4 hr but increased fructose (6 hr) in the resting phase. In the preparatory phase (active oogenesis), fructose decreased significantly at 4 hr and glucose at 6 and 12 hr. In the prespawning phase, HCG administration induced a significant decrease in glucose level at 2 hr but increased it significantly at 16 and 18 hr. Fructose concentration, on the other hand, decreased > or = 4 hr. The involvement of different energy substrates (glucose, fructose, 2-deoxyglucose and pyruvate) on final oocyte maturation and viability of eggs were demonstrated in an in vitro study, employing incubation of mature oocytes in medium containing one or the other energy substrates. Glucose at 4 and 8 mM concentrations increased the viability of eggs without affecting germinal vesicle break down (GVBD) and at high concentrations (16 and 32 mM) marginally increased GVBD but decreased viability. Pyruvate at high concentrations (> or = 8 mM) produced a positive effect on both GVBD and viability. Fructose and 2-deoxyglucose did not elicit any effect on these parameters. The results suggest that glucose has an active role in catfish oogenesis and maintaining egg quality and HCG influences monosaccharide metabolism in a season-dependent manner.

Valor preditivo da concentração sérica de hCG no 14º dia após transferência embrionária na evolução da gestação / The predictive value of Hcg serum concentration on the 14th day after embryo transfer in pregnancy outcome

OBJETIVO: Avaliar se uma única dosagem sérica de hCG no 14º dia após transferência embrionária é capaz depredizer a evolução da gestação. MATERIAIS E MÉTODOS: Foram avaliadas retrospectivamente 41 pacientes submetidas a FIV ou ICSI que apresentavam teste de gravidez positivo (hCG > 25 mUI/mL) no 14° dia após transferência embrionária. As pacientes foram divididas em dois grupos: gestação não viável (gestações químicas, prenhezes ectópicas e abortos no 1° trimestre) e gestação viável (abortos no 2° trimestre, gestações únicas ou múltiplas). RESULTADOS: A concentração média de hCG nas gestações viáveis (550,6 mUI/mL) foi maior do que a das gestações não viáveis (179 mUI/mL). Nas gestações viáveis múltiplas a concentração média de hCG foi quase três vezes maior do que a observada nas gestações viáveis únicas (875,3 mUI/mL vs 334,1 mUI/mL). A concentração média de hCG nas gestações não viáveis não apresentou diferença significativa daquela encontrada nas gestações viáveis únicas, no entanto as gestações químicas apresentaram menor concentração de hCG quando comparadas as gestações únicas viáveis (123,9 mUI/mL vs 334,1 mUI/mL). De acordo com a curva ROC, pacientes que apresentam concentração sérica de hCG maior do que 125 mUI/mL, têm 88,2 de probabilidade de evoluir com gestação viável. CONCLUSÃO: Uma única dosagem sérica de hCG no 14° dia após transferência embrionária apresenta bom valor preditivo na determinação da evolução da gestação

Response related enzymatic changes in ovaries of superovulated mice.

Adult female mice were superovulated with PMSG followed by HCG and 140 blastocysts and 69 morulae were recovered from 24 mice. On the basis of the response, mice were divided into six groups; non responders, 1-5, 6-10, 11-20, 21-30 and >30 embryos. The ovaries of the animals were pooled group wise, homogenized in PBS (pH 7.4) and after centrifugation for 10-15 minutes, the supernatant was analyzed for the enzymes, guanine oxaloacetate transaminase (GOT), guanine pymvate transaminase (GPT), acid phosphatases (ACP) and alkaline phosphatases (AKP). Acid and alkaline phosphatase activities did not show any variation in relation to response to superovulation but GOT and GPT showed significantly increased activity in response to induction of superovulation. A statistically significant positive correlation was found between GOT and GPT activities and the superovulatory response in mice.

Disgenesia gonadal mixta: ambigüedad genital. Urgencias y problemas diagnósticos / Mixed gonadal dysgenesia: genital ambigüity. Emergency and diagnostic problems

Presentamos un caso de Disgenesia Gonadal Mixta que anátomicamente presenta: ambigüedad de genitales externos con seno urogenital al que desemboca vagina, útero y trompas normales. La gónada derecha corresponde a testículo con extensas áreas de fibrosis y la izquierda a una gónada disgenética. La cromatina sexual fue positiva, también lo fue el Test de Hibridación in situ para la región centromérica de Cromosoma Y. El cariotipo determinó un mosaicismo 45 X (41 por ciento de las metafases estudiadas) / 46 X + marc (?Y). Se decidió la extirpación quirúrgica de ambas gónadas.