RESUMO
Endometrium extracellular matrix provides a wide range of signals at different cellular levels, like cell death and proliferation, which can be important for regulating menses and reducing the proliferative processes. The objective of this study is to evaluate hyaluronic acid concentration, the enzymes of hyaluronic acid synthases in the endometrium of patients with polycystic ovary syndrome (PCOS) and eumenorrheic women. A total of 60 endometrial samples from 30 patients with PCOS and 30 women with regular menstrual cycles in the proliferative phase, attended at Gynecology Division of Clinical Hospital of the FMUSP (HC-USP). Profile determination and the concentration of hyaluronic acid was performed by the biochemical method of the fluorimetric assay (ELISA-like). Its location in the endometrial tissue as well as the dosage of enzymes synthases (HAS1, HAS2 and HAS3) was done by immunohistochemistry and western blotting. Statistical analyses were performed with one-way ANOVA, followed by the Bonferroni test. Regarding hyaluronic acid synthases, there was a higher HAS1 and HAS2 reactivity and lower HAS3 reactivity in the PCOS endometrium compared to women with regular menstrual cycles in the proliferative phase. We suggest that PCOS patients have different composition of hyaluronic acid in relation to a regular cycle in the proliferative phase.
Assuntos
Endométrio/metabolismo , Fase Folicular/metabolismo , Hialuronan Sintases/metabolismo , Ácido Hialurônico/metabolismo , Síndrome do Ovário Policístico/metabolismo , Adulto , Estudos de Casos e Controles , Feminino , Humanos , Projetos Piloto , Adulto JovemRESUMO
When levonorgestrel (LNG) is given for emergency contraception during the follicular phase, it not only inhibits or delays ovulation, but also induces changes in endometrial secretions that modulate sperm functionality. In order to characterize the female reproductive tract secreted molecules that may affect human spermatozoa, we analyzed changes in the protein content of uterine flushings obtained from women during the periovulatory phase of a control and a LNG-treated menstrual cycle. Lectin affinity analysis and 2D gel electrophoresis of uterine samples showed changes in protein glycosylation patterns and the presence of 31 differentially expressed proteins (8 upregulated and 23 downregulated). Mass spectrometry and Western blot analyses of the differential expressed proteins showed lactotransferrin (LTF) as one of the upregulated molecules by LNG. In this study, LTF exhibited significant dose-related effects on sperm functionality, particularly a decrease of calcium ionophore-induced acrosome reaction and protein tyrosine phosphorylation. Overall, the results indicated that LNG promoted changes in the proteome of uterine secretions that might compromise human sperm capacitation. These data further support the participation of other mechanisms of action of LNG as emergency contraceptive, in addition to those on ovulation.
Assuntos
Contraceptivos Hormonais/uso terapêutico , Fase Folicular/efeitos dos fármacos , Lactoferrina/metabolismo , Lactoferrina/farmacologia , Levanogestrel/uso terapêutico , Espermatozoides/efeitos dos fármacos , Útero/efeitos dos fármacos , Reação Acrossômica/efeitos dos fármacos , Adulto , Ionóforos de Cálcio/farmacologia , Feminino , Fase Folicular/metabolismo , Glicosilação , Humanos , Masculino , Ovulação/efeitos dos fármacos , Fosforilação , Espermatozoides/metabolismo , Tirosina/metabolismo , Útero/metabolismo , Adulto JovemRESUMO
Culture systems are available for human granulosa cells (GCs) that perpetuate luteinization. The present study examines the plating density effects and long-term serum-free culture on the in vitro dynamics differentiation of luteinizing human GCs. Cells were cultured in serum-free α-minimum essential medium (α-MEM) or serum-based tissue culture medium (TCM). The time course of GCs morphology and secretion of estradiol (E2), progesterone (P4), and relaxin were analyzed after 48, 96, and 144 hours of culture. Other functional markers as follicle-stimulating hormone/luteinizing hormone receptors and steroidogenic enzymes were investigated at the end of culture. The morphology of an α-MEM cell rather than a TCM cell resembles more closely that seen in vivo. Compared to TCM cultures, α-MEM cells secreted 93.7% and 87.2% more E2 and approximately 7% and 17% of the amount of P4 when cultured at densities of 2 × 10(4) or 4 × 10(4) cells/well, respectively. Relaxin secretion was significantly reduced in α-MEM cultures. α-MEM cells were estrogenic and expressed the CYP19 gene. Levels of CYP17 increased about 8-fold in α-MEM cells above the levels found in TCM cells. Our results reveal new insights into human GCs differentiation in vitro and demonstrate the critical importance of the culture system and cell-plating density on the establishment of estrogenic or progestogenic GC phenotypes.
Assuntos
Diferenciação Celular , Meios de Cultura Livres de Soro/metabolismo , Fertilização in vitro , Fase Folicular/metabolismo , Células Lúteas/metabolismo , Aromatase/biossíntese , Aromatase/genética , Forma Celular , Sobrevivência Celular , Células Cultivadas , Indução Enzimática , Estradiol/metabolismo , Feminino , Humanos , Fenótipo , Progesterona/metabolismo , Relaxina/metabolismo , Fatores de TempoRESUMO
The aim of this study was to quantify the sulfated glycosaminoglycans in the endometria of women with polycystic ovary syndrome (PCOS). Of the 18 patients recruited for this study, 10 patients with PCOS comprised the PCOS group (PCOSG), and eight patients with regular and ovulatory menstrual cycles comprised the control group (CG). The clinical, biochemical, morphological and endometrial data from both groups were analyzed. Biopsies were performed during the proliferative phase of the menstrual cycle for the CG and during the persistent proliferative phase for the PCOSG (all women were amenorrheic). In the PCOSG, there was a significant increase in the endometrial concentration levels of heparan sulfate (p = 0.03), but no difference in the concentrations of chondroitin sulfate was determined between the two groups (p = 0.77). Period of time without menstruation (p = 0.001) and body mass index (BMI) (p = 0.04) correlated directly and positively with heparan sulfate concentration. There was no association between heparan sulfate levels and basal insulin values (p = 0.08). High levels of endometrial heparan sulfate in women with PCOS indicate an interference with maternal-fetal recognition, which contributes to infertility; thus, endometrial heparan sulfate may be a predictive marker of future neoplasia risk.
Assuntos
Endométrio/metabolismo , Heparitina Sulfato/metabolismo , Infertilidade Feminina/etiologia , Síndrome do Ovário Policístico/metabolismo , Regulação para Cima , Adulto , Biomarcadores/metabolismo , Biópsia , Índice de Massa Corporal , Brasil/epidemiologia , Sulfatos de Condroitina/metabolismo , Neoplasias do Endométrio/epidemiologia , Endométrio/patologia , Feminino , Fase Folicular/metabolismo , Hospitais Universitários , Humanos , Ambulatório Hospitalar , Sobrepeso/complicações , Síndrome do Ovário Policístico/complicações , Síndrome do Ovário Policístico/patologia , Síndrome do Ovário Policístico/fisiopatologia , Risco , Adulto JovemRESUMO
Although superstimulatory protocols in cattle are usually initiated during mid-cycle, the elective control follicular wave emergence and ovulation have had a great impact on the application of on-farm embryo transfer. However, the most commonly used approach for the sinchronization of follicular wave emergence invoolves the use of estradiol which cannot be used in many parts of the world. Therefore, the need for alternative treatments has driven recent research. An approach that has shown promise is to initiate FSH treatments at the time of the emergence of the first follicular wave folliwing GnRH-induced ovulation. Alternatively, it has been shown that that it may be possible to ignore follicular wave status, and by extending the treatment protocol induce smaller follicles to grow and reach maturity and superovulate. Finally , the short half-life of pintuitary FSH necessitates twice daily treatments which are times consuming, stressful and subject to error. Recent treatment protocols have permitted supertimulation with a single or alternatively two-FSH treatments, reducing the need for animal handling during FSH treatments.(AU)
Assuntos
Animais , Embrião de Mamíferos/embriologia , Estradiol/farmacologia , Ovulação/metabolismo , Bovinos/classificação , Fase Folicular/metabolismoRESUMO
Although superstimulatory protocols in cattle are usually initiated during mid-cycle, the elective control follicular wave emergence and ovulation have had a great impact on the application of on-farm embryo transfer. However, the most commonly used approach for the sinchronization of follicular wave emergence invoolves the use of estradiol which cannot be used in many parts of the world. Therefore, the need for alternative treatments has driven recent research. An approach that has shown promise is to initiate FSH treatments at the time of the emergence of the first follicular wave folliwing GnRH-induced ovulation. Alternatively, it has been shown that that it may be possible to ignore follicular wave status, and by extending the treatment protocol induce smaller follicles to grow and reach maturity and superovulate. Finally , the short half-life of pintuitary FSH necessitates twice daily treatments which are times consuming, stressful and subject to error. Recent treatment protocols have permitted supertimulation with a single or alternatively two-FSH treatments, reducing the need for animal handling during FSH treatments.
Assuntos
Animais , Embrião de Mamíferos/embriologia , Estradiol/farmacologia , Ovulação/metabolismo , Bovinos/classificação , Fase Folicular/metabolismoRESUMO
OBJECTIVE: To evaluate nuclear factor kappaB (NF-κB) activation and NF-κB-p65 subunit activation, immunolocalization, and expression in the endometrium of healthy women and endometriosis patients throughout the menstrual cycle. DESIGN: Prospective observational study. SETTING: Affiliated hospital and university research laboratory. PATIENT(S): Twenty-four healthy women and 24 endometriosis patients. INTERVENTION(S): Menstrual, proliferative, and secretory endometrial biopsies. MAIN OUTCOME MEASURE(S): Assessment of NF-κB and p65 activation by protein-DNA binding assays and p65 localization and expression by immunohistochemistry. RESULT(S): Total NF-κB-DNA binding was constitutive and variable in human endometrium accross the menstrual cycle. Healthy women (physiologic conditions) showed higher p65-DNA binding in proliferative than in menstrual and secretory endometrium. Conversely, in endometriosis patients, p65-DNA binding was higher in proliferative and secretory endometrium than in menstrual endometrium. Endometrial epithelial cells showed higher p65 expression level score than endometrial stromal cells. CONCLUSION(S): NF-κB activity is constitutive, physiologic, and variable in human endometrium. The physiologic cyclic p65 activation pattern was altered in endometriosis patients, showing no cyclic variation between the proliferative and secretory phase of the menstrual cycle. The absence of decreased p65 activity in secretory endometrium from endometriosis patients is concurrent with progesterone resistance and could participate in endometrial biologic alterations during the implantation window in endometriosis patients.
Assuntos
Endometriose/metabolismo , Endométrio/metabolismo , Ciclo Menstrual/metabolismo , NF-kappa B/metabolismo , Adulto , Sítios de Ligação , Biópsia , Estudos de Casos e Controles , Chile , DNA/metabolismo , Ensaio de Desvio de Mobilidade Eletroforética , Endometriose/patologia , Endométrio/patologia , Células Epiteliais/metabolismo , Feminino , Fase Folicular/metabolismo , Humanos , Imuno-Histoquímica , Fase Luteal/metabolismo , Menstruação/metabolismo , Pessoa de Meia-Idade , Estudos Prospectivos , Células Estromais/metabolismo , Fator de Transcrição RelA/metabolismo , Adulto JovemRESUMO
BACKGROUND: The purpose of the study was to determine if the effect of llama OIF on LH secretion is mediated by stimulation of the hypothalamus or pituitary gland. METHODS: Using a 2-by-2 factorial design to examine the effects of OIF vs GnRH with or without a GnRH antagonist, llamas with a growing ovarian follicle greater than or equal to 8 mm were assigned randomly to four groups (n = 7 per group) and a) pre-treated with 1.5 mg of GnRH antagonist (cetrorelix acetate) followed by 1 mg of purified llama OIF, b) pre-treated with 1.5 mg of cetrorelix followed by 50 micrograms of GnRH, c) pre-treated with a placebo (2 ml of saline) followed by 1 mg of purified llama OIF or d) pre-treated with a placebo (2 ml of saline) followed by 50 micrograms of GnRH. Pre-treatment with cetrorelix or saline was given as a single slow intravenous dose 2 hours before intramuscular administration of either GnRH or OIF. Blood samples for LH measurement were taken every 15 minutes from 1.5 hours before to 8 hours after treatment. The ovaries were examined by ultrasonography to detect ovulation and CL formation. Blood samples for progesterone measurement were taken every-other-day from Day 0 (day of treatment) to Day 16. RESULTS: Ovulation rate was not different (P = 0.89) between placebo+GnRH (86%) and placebo+OIF groups (100%); however, no ovulations were detected in llamas pre-treated with cetrorelix. Plasma LH concentrations surged (P < 0.01) after treatment in both placebo+OIF and placebo+GnRH groups, but not in the cetrorelix groups. Maximum plasma LH concentrations and CL diameter profiles did not differ between the placebo-treated groups, but plasma progesterone concentrations were higher (P < 0.05), on days 6, 8 and 12 after treatment, in the OIF- vs GnRH-treated group. CONCLUSION: Cetrorelix (GnRH antagonist) inhibited the preovulatory LH surge induced by OIF in llamas suggesting that LH secretion is modulated by a direct or indirect effect of OIF on GnRH neurons in the hypothalamus.
Assuntos
Camelídeos Americanos , Fase Folicular/efeitos dos fármacos , Hormônio Liberador de Gonadotropina/análogos & derivados , Hormônio Luteinizante/metabolismo , Ovulação/efeitos dos fármacos , Animais , Camelídeos Americanos/sangue , Camelídeos Americanos/metabolismo , Camelídeos Americanos/fisiologia , Regulação para Baixo/efeitos dos fármacos , Feminino , Fármacos para a Fertilidade/metabolismo , Fármacos para a Fertilidade/farmacologia , Fase Folicular/metabolismo , Hormônio Liberador de Gonadotropina/farmacologia , Antagonistas de Hormônios/farmacologia , Hormônio Luteinizante/sangue , Masculino , Indução da Ovulação/métodos , Placebos , Fluxo Pulsátil/efeitos dos fármacos , Sêmen/metabolismo , Sêmen/fisiologia , Proteínas de Plasma Seminal/metabolismo , Proteínas de Plasma Seminal/farmacologiaRESUMO
PURPOSE: To assess the potential effects of short-term exposure to particulate air pollution during follicular phase on clinical, laboratory, and pregnancy outcomes of women undergoing IVF/ET. METHODS: Retrospective cohort study of 400 first IVF/ET cycles of women exposed to ambient particulate matter during follicular phase. Particulate matter (PM) was categorized into quartiles (Q(1): < or =30.48 microg/m(3), Q(2): 30.49-42.00 microg/m(3), Q(3): 42.01-56.72 microg/m(3), and Q(4): >56.72 microg/m(3)). RESULTS: Clinical, laboratory, or treatment variables were not affected by follicular phase PM exposure periods. Women exposed to Q(4) period during the follicular phase of conception cycles had a higher risk of miscarriage (odds ratio, 5.05; 95% confidence interval: 1.04-25.51) when compared to women exposed to Q(1-3) periods. CONCLUSION: Our results show an association between brief exposure to high levels of ambient PM during the preconceptional period and early pregnancy loss, although no effect of this exposure on clinical, laboratory, and treatment outcomes was observed.
Assuntos
Transferência Embrionária , Fertilização in vitro , Exposição Materna/efeitos adversos , Material Particulado/toxicidade , Resultado da Gravidez , Aborto Espontâneo/etiologia , Estudos de Coortes , Feminino , Fertilização , Fase Folicular/metabolismo , Humanos , Infertilidade Feminina/terapia , Gravidez , Estudos RetrospectivosRESUMO
OBJECTIVE: To study the protein profile of oocytes and cumulus cells from different sized follicles throughout the follicular phase and to asses the ability of oocytes to progress from the dictyate to metaphase II (MII) stage. DESIGN: Animal model study. SETTING: Five academic basic research laboratories and the National Primate Centre. ANIMAL(S): Eleven normal, cycling capuchin monkey (Cebus apella) females. INTERVENTION(S): Cumulus-oocyte complexes and denuded oocytes were recovered by antral follicle aspiration. MAIN OUTCOME MEASURE(S): Protein profile analysis of denuded or intact oocytes. RESULT(S): The protein profiles of 25 denuded or intact oocytes recovered on days 5 (six denuded, five intact), 7 (four denuded, four intact), or 9 (one denuded, five intact) of the menstrual cycle were analyzed; in a second experiment, 40 intact oocytes were cultured for 24 (n = 20) or 36 hours (n = 20). The oocytes were denuded, fixed, stained, and microscopically examined to reveal the meiotic stage. The protein profile in each compartment within the cumulus-oocyte complex varied along the follicular development with a predominance of low-molecular-weight proteins in both oocyte and cumulus cells at final stages. No differences were found in the protein profile among oocytes pertaining to different sized follicles that were in the same day of the follicular phase. Oocyte MII competence was achieved only after incubation for 36 hours, and the highest maturation rate occurred in those becoming from dominant follicles. CONCLUSION(S): Our study shows, for the first time in a New World primate species, that the proteins contained in oocytes and cumulus cells reach an identical profile in the late follicular phase. This phenomenon could be related to the oocyte's ability to progress to the MII stage.
Assuntos
Cebus , Células do Cúmulo/metabolismo , Fase Folicular/metabolismo , Meiose , Oócitos/metabolismo , Proteínas/metabolismo , Animais , Células Cultivadas , Feminino , Fatores de TempoRESUMO
OBJECTIVES: Fibroadenoma is the most common benign mammary condition among women aged 35 or younger. Expression of Ki-67 antigen has been used to compare proliferative activity of mammary fibroadenoma epithelium in the follicular and luteal phases of the menstrual cycle. MATERIALS AND METHODS: Ninety eumenorrheic women were selected for tumour excision; they were assigned to either of the two groups, according to their phase of menstrual cycle. At the end of the study, 75 patients with 87 masses were evaluated by epithelial cell Ki-67 expression, blind (no information given concerning group to which any lesion belonged). RESULTS: Both groups were found to be homogeneous relative to age, menarche, body mass index, previous gestation, parity, breastfeeding, number of fibroadenomas, family history of breast cancer and tabagism. Median tumour size was 2.0 cm and no relationship between proliferative activity and nodule diameter was observed. No typical pattern was observed in the expression of Ki-67 in distinct nodules of the same patient. Average values for expression of Ki-67 (per 1000 epithelial cells) in follicular and luteal phases were 27.88 and 37.88, respectively (P = 0.116). CONCLUSION: Our findings revealed that proliferative activities in the mammary fibroadenoma epithelium did not present a statistically significant difference in the follicular and luteal phases. The present study contributes to clarifying that fibroadenoma is a neoplasm and does not undergo any change in the proliferative activity during the menstrual cycle.
Assuntos
Neoplasias da Mama/patologia , Proliferação de Células , Fibroadenoma/patologia , Fase Folicular/metabolismo , Antígeno Ki-67/metabolismo , Fase Luteal/metabolismo , Adolescente , Adulto , Fatores Etários , Aleitamento Materno/estatística & dados numéricos , Neoplasias da Mama/complicações , Neoplasias da Mama/metabolismo , Feminino , Fibroadenoma/complicações , Fibroadenoma/metabolismo , Fase Folicular/sangue , Humanos , Fase Luteal/sangue , Obesidade/complicações , Progesterona/sangue , Tabagismo/complicações , Adulto JovemRESUMO
Foram avaliadas quatro técnicas de sincronização da onda folicular em protocolos de superovulação. Para tal, foram utilizadas 112 vacas doadoras, das raças Simental, Limousin e Red Angus, com escore corporal médio de 3,0. Os animais foram divididos aleatoriamente em cinco grupos experimentais de acordo com o método de sincronização da emergência da onda folicular. Foram realizadas 30 superovulações em cada grupo, considerando os seguintes protocolos: GI - grupo controle animais superovulados entre o 8o e o 12o dia do ciclo estral (dia zero = estro); GII animais que sofreram punção folicular no 9o dia (dia 0 = estro) e início do tratamento superovulatório no 11o dia; GIII animais que sofreram punção folicular em fase não conhecida do ciclo estral, associada ao uso de um dispositivo intravaginal contendo progesterona (P4) e tratamento superovulatório iniciado 48h após a punção, GIV animais que utilizaram implante intravaginal de progesterona colocadoem fase aleatória do ciclo estral, mantido por nove dias, associado à administração de 50 mg de P4 e de 2mg de benzoato de estradiol, sendo o tratamento superovulatório iniciado cinco dias após a colocação do dispositivo e GV animais que receberam implante intravaginal de P4 colocado em fase aleatória do ciclo estral e mantido por oito dias, associado à administração de 50mg de P4 e 2mg de 17â-estradiol, sendo o tratamento superovulatório iniciado quatro dias após a colocação do dispositivo. Nos grupos I, II, III, IV e V o total de estruturas coletadas e de embriões viáveis foram, respectivamente (13,53±9,23 vs 13,87 ± 7,85 vs 18,70 ± 10,88 vs 9,03 ± 4,97 vs 13,60 ± 8,39) e (8,43±5,68 vs 8,27 ± 7,06 vs 10,47 ± 8,19 vs 5,37 ± 2,92 vs 7,23 ± 5,30). Os resultados observados no GIII foram superiores ao GI, GII, GIV e GV (P< 0,05), respectivamente. Os grupos I, II e IV não apresentaram diferenças entre si (P> 0,05), enquanto o desempenho de GIV foi inferior (P<0,05). Os resultados permitem concluir que é possível sincronizar a emergência da onda folicular de vacas doadoras, com início da superovulação em qualquer momento do ciclo estral, e que o tratamento progestágeno associado à punção folicular oferece os melhores resultados.(AU)
We evaluated four different techniques of follicular wave synchronization by comparing total number of structures recovery, number of viable and degenerated embryos, number of oocytes recovery and cost of uterine flushing. One hundred twelve Simental, Limousin and Red Angus donators were randomly allocated in five treatment groups according to protocol used for follicular wave emergence synchronization. Thirty superovulations were performed in each group, the programs were: G1- Control, superovulation treatment between days 8 and 12 of the estrous cycle; G2- follicle aspiration on day 9 of the estrous cycle; G3- follicle aspiration plus intravaginal progesterone implant; G4- intravaginal progesterone implant plus estradiol benzoate and G5- intravaginal progesterone implant plus estradiol-17â. Artificial insemination was performed twice, 12 and 24 hours after detection of behavioral estrus. Embryo collection was performed on day 7 after inseminations. Total number of recovered structures (18,70 ± 10,88 vs 13,53 ± 9,23) and viable embryos (10,47 ± 8,19 vs 8,43 ± 5,68) were higher (P< 0,05) in G3 than in G1, respectively. No differences (P>0,05) were detected amongst groups 2, 5 and 1, while performance of G4 was the lowest (P<0,05). Results demonstrate to be possible synchronize follicular wave emergence by initiating superstimulation at any time of the estrous cycle. Additionally we verified that the program using progesterone associated to follicular ablation had the best results in synchronizing follicular wave emergence aiming superstimulation in cattle.(AU)
Assuntos
Animais , Feminino , Bovinos , Fase Folicular/metabolismo , Superovulação/genética , Bovinos/classificação , Tratamento Biológico/métodos , HormôniosRESUMO
Foram avaliadas quatro técnicas de sincronização da onda folicular em protocolos de superovulação. Para tal, foram utilizadas 112 vacas doadoras, das raças Simental, Limousin e Red Angus, com escore corporal médio de 3,0. Os animais foram divididos aleatoriamente em cinco grupos experimentais de acordo com o método de sincronização da emergência da onda folicular. Foram realizadas 30 superovulações em cada grupo, considerando os seguintes protocolos: GI - grupo controle animais superovulados entre o 8o e o 12o dia do ciclo estral (dia zero = estro); GII animais que sofreram punção folicular no 9o dia (dia 0 = estro) e início do tratamento superovulatório no 11o dia; GIII animais que sofreram punção folicular em fase não conhecida do ciclo estral, associada ao uso de um dispositivo intravaginal contendo progesterona (P4) e tratamento superovulatório iniciado 48h após a punção, GIV animais que utilizaram implante intravaginal de progesterona colocadoem fase aleatória do ciclo estral, mantido por nove dias, associado à administração de 50 mg de P4 e de 2mg de benzoato de estradiol, sendo o tratamento superovulatório iniciado cinco dias após a colocação do dispositivo e GV animais que receberam implante intravaginal de P4 colocado em fase aleatória do ciclo estral e mantido por oito dias, associado à administração de 50mg de P4 e 2mg de 17â-estradiol, sendo o tratamento superovulatório iniciado quatro dias após a colocação do dispositivo. Nos grupos I, II, III, IV e V o total de estruturas coletadas e de embriões viáveis foram, respectivamente (13,53±9,23 vs 13,87 ± 7,85 vs 18,70 ± 10,88 vs 9,03 ± 4,97 vs 13,60 ± 8,39) e (8,43±5,68 vs 8,27 ± 7,06 vs 10,47 ± 8,19 vs 5,37 ± 2,92 vs 7,23 ± 5,30). Os resultados observados no GIII foram superiores ao GI, GII, GIV e GV (P 0,05), enquanto o desempenho de GIV foi inferior (P<0,05). Os resultados permitem concluir que é possível sincronizar a emergência da onda folicular de vacas doadoras, com início da superovulação em qualquer momento do ciclo estral, e que o tratamento progestágeno associado à punção folicular oferece os melhores resultados.
We evaluated four different techniques of follicular wave synchronization by comparing total number of structures recovery, number of viable and degenerated embryos, number of oocytes recovery and cost of uterine flushing. One hundred twelve Simental, Limousin and Red Angus donators were randomly allocated in five treatment groups according to protocol used for follicular wave emergence synchronization. Thirty superovulations were performed in each group, the programs were: G1- Control, superovulation treatment between days 8 and 12 of the estrous cycle; G2- follicle aspiration on day 9 of the estrous cycle; G3- follicle aspiration plus intravaginal progesterone implant; G4- intravaginal progesterone implant plus estradiol benzoate and G5- intravaginal progesterone implant plus estradiol-17â. Artificial insemination was performed twice, 12 and 24 hours after detection of behavioral estrus. Embryo collection was performed on day 7 after inseminations. Total number of recovered structures (18,70 ± 10,88 vs 13,53 ± 9,23) and viable embryos (10,47 ± 8,19 vs 8,43 ± 5,68) were higher (P0,05) were detected amongst groups 2, 5 and 1, while performance of G4 was the lowest (P<0,05). Results demonstrate to be possible synchronize follicular wave emergence by initiating superstimulation at any time of the estrous cycle. Additionally we verified that the program using progesterone associated to follicular ablation had the best results in synchronizing follicular wave emergence aiming superstimulation in cattle.
Assuntos
Feminino , Animais , Bovinos , Fase Folicular/metabolismo , Superovulação/genética , Bovinos/classificação , Hormônios , Tratamento Biológico/métodosRESUMO
The menstrual cycle has been pointed out as a factor influencing halitosis. However, this relationship has not yet been clarified. The aim of this study was to evaluate the influence of gender and the menstrual cycle on the production of volatile sulphur compounds (VSC) in women (n=14) across the menstrual cycle, and in men (n=17). Volunteers in good oral and general health were submitted to the evaluation of VSC, salivary flow, cortisol and anaerobic bacteria counts in saliva. Data were compared among groups by Analysis of Variance (alpha=5%). VSC was higher in the menstrual and premenstrual phases when compared with men and the follicular phase (p<0.05). Salivary flow was lower in the menstrual and premenstrual phases when compared with men and the follicular phase (p<0.05). Salivary cortisol was higher in the menstrual phase in comparison with men and the premenstrual and follicular phases (p<0.05). Total salivary protein was higher in men when compared to women (p<0.05) with no differences among menstrual phases (p>0.05). Levels of anaerobic micro-organisms, however, were not different among groups (p>0.05). In conclusion, the production of VSC is influenced by menstrual cycle and protein concentration and salivary flow might be involved in this process.
Assuntos
Halitose/etiologia , Ciclo Menstrual/metabolismo , Saliva/química , Compostos de Enxofre/metabolismo , Análise de Variância , Feminino , Fase Folicular/metabolismo , Halitose/microbiologia , Humanos , Masculino , Saliva/microbiologia , Taxa Secretória/fisiologia , Fatores Sexuais , Estudantes de Medicina , Inquéritos e Questionários , Adulto JovemRESUMO
Avaliou-se o efeito do diâmetro e da fase do desenvolvimento folicular sobre a competência de oócitos para a produção in vitro de embriões bovinos. A primeira onda folicular foi sincronizada com progestógeno por nove dias e 24 horas após a sua retirada aplicou-se LH. Os ovários foram recuperados 60h (G-60), 96h (G-96) e 108h (G-108) após a ovulação induzida pelo LH. Os folículos foram dissecados ou aspirados e medidos e os oócitos recuperados e submetidos à maturação, fecundação e cultivo in vitro. Os ovários do G-60 apresentaram mais oócitos viáveis (graus I, II e III) (96,6 por cento). A taxa de clivagem teve efeito significativo sobre o diâmetro folicular, sendo maior nos oócitos oriundos de folículos classe 3 (>7mm). Na taxa de produção de blastocisto observou-se interação diâmetro versus fase de desenvolvimento folicular. A taxa de produção de blastocisto foi maior em oócitos obtidos de folículos com diâmetros <5mm (classe 1) no G-60 (64,5 por cento), de 5-7mm (classe 2) no G-96 (33,3 por cento) e >7mm (classe 3) no G-108 (50 por cento). Conclui-se que o diâmetro e a fase de desenvolvimento folicular influenciam a competência oocitária para o desenvolvimento in vitro. Nos estádios iniciais da onda folicular a produção de blastocisto foi maior em oócitos de folículos pequenos; com o avanço da onda, a produção de blastocistos foi maior em oócitos obtidos de folículos maiores.
The effect of follicular phase and follicle diameter on in vitro production of bovine embryos was evaluated. Follicular wave was synchronized in Nelore heifers with a progestogen for nine days and LH was administered 24 hours after progestogen withdrawal. Ovaries were recovered 60h (G-60), 96 h (G-96), or 108 h (G-108) after LH treatment. Ovarian follicles were dissected or aspirated and measured before oocytes were recovered and submitted to in vitro maturation, fertilization, and culture. The G-60 ovaries presented more viable oocyte (degrees I, II and III) (96.6 percent). Cleavage rate was higher for oocytes from follicles 7mm in diameter (class 3). There was a follicular phase-by-follicle diameter interaction effect on blastocyst production rate. Blastocyst production rates were higher for oocytes from follicles 5mm in diameter (class 1) in the G-60 group (64.5 percent), from follicles 5-7mm (class 2) in the G-96 group (33.3 percent), and from follicles 7mm (class 3) in the G-108 group (50 percent). Both the phase of follicular development and the follicle diameter influenced oocyte competence and ability for development in vitro. At the initial stages of the follicular wave, blastocyst production was higher for oocytes from small follicles. However, as the follicular wave advances, blastocyst production increases for oocytes from larger follicles.
Assuntos
Animais , Feminino , Blastocisto/fisiologia , Bovinos , Estruturas Embrionárias/fisiologia , Fase Folicular/metabolismo , Oócitos/fisiologiaRESUMO
Avaliou-se o efeito do diâmetro e da fase do desenvolvimento folicular sobre a competência de oócitos para a produção in vitro de embriões bovinos. A primeira onda folicular foi sincronizada com progestógeno por nove dias e 24 horas após a sua retirada aplicou-se LH. Os ovários foram recuperados 60h (G-60), 96h (G-96) e 108h (G-108) após a ovulação induzida pelo LH. Os folículos foram dissecados ou aspirados e medidos e os oócitos recuperados e submetidos à maturação, fecundação e cultivo in vitro. Os ovários do G-60 apresentaram mais oócitos viáveis (graus I, II e III) (96,6 por cento). A taxa de clivagem teve efeito significativo sobre o diâmetro folicular, sendo maior nos oócitos oriundos de folículos classe 3 (>7mm). Na taxa de produção de blastocisto observou-se interação diâmetro versus fase de desenvolvimento folicular. A taxa de produção de blastocisto foi maior em oócitos obtidos de folículos com diâmetros <5mm (classe 1) no G-60 (64,5 por cento), de 5-7mm (classe 2) no G-96 (33,3 por cento) e >7mm (classe 3) no G-108 (50 por cento). Conclui-se que o diâmetro e a fase de desenvolvimento folicular influenciam a competência oocitária para o desenvolvimento in vitro. Nos estádios iniciais da onda folicular a produção de blastocisto foi maior em oócitos de folículos pequenos; com o avanço da onda, a produção de blastocistos foi maior em oócitos obtidos de folículos maiores.(AU)
The effect of follicular phase and follicle diameter on in vitro production of bovine embryos was evaluated. Follicular wave was synchronized in Nelore heifers with a progestogen for nine days and LH was administered 24 hours after progestogen withdrawal. Ovaries were recovered 60h (G-60), 96 h (G-96), or 108 h (G-108) after LH treatment. Ovarian follicles were dissected or aspirated and measured before oocytes were recovered and submitted to in vitro maturation, fertilization, and culture. The G-60 ovaries presented more viable oocyte (degrees I, II and III) (96.6 percent). Cleavage rate was higher for oocytes from follicles 7mm in diameter (class 3). There was a follicular phase-by-follicle diameter interaction effect on blastocyst production rate. Blastocyst production rates were higher for oocytes from follicles 5mm in diameter (class 1) in the G-60 group (64.5 percent), from follicles 5-7mm (class 2) in the G-96 group (33.3 percent), and from follicles 7mm (class 3) in the G-108 group (50 percent). Both the phase of follicular development and the follicle diameter influenced oocyte competence and ability for development in vitro. At the initial stages of the follicular wave, blastocyst production was higher for oocytes from small follicles. However, as the follicular wave advances, blastocyst production increases for oocytes from larger follicles.(AU)
Assuntos
Animais , Feminino , Oócitos/fisiologia , Fase Folicular/metabolismo , Blastocisto/fisiologia , Estruturas Embrionárias/fisiologia , BovinosRESUMO
The relative proportion of the circulating luteinizing hormone isoforms in goats during follicular phase (pre-ovulatory peak; F) and anestrus (A) was investigated. Estrus was synchronized in six goats with a prostaglandin analogue. After estrus was detected, blood samples were taken at 1 h intervals for 24 h. Four anestrous goats received 100 microg i.v. of GnRH and blood samples were collected every 15 min for 5 h. Samples with the greatest LH concentration in follicular phase and after GnRH administration (anestrus) were analyzed by chromatofocusing and eluted with a pH gradient from 10.5 to 3.5. For quantification purposes eluted LH was grouped into basic (pH> or =7.5), neutral (pH 7.4-6.5) and acidic isoforms (pH< or =6.4) as well as by pH unit. In both physiological conditions (PC), basic and acidic isoforms were greater than the neutral. With this grouping criteria, there was an interaction between PC and pH group, with the proportion of neutral isoforms being greater (p<0.05) in A (12.0+/-0.8%) as compared with F (5+/-2%). Analysis by pH unit showed a very basic group of eluted isoforms (pH> or =10), which amounted to a percentage of 6.0+/-0.4% of the total observed during A, and 3+/-1% during F (p<0.05). Predominant isoforms in A eluted in the pH range 9.99-9.0 (42+/-3%) as compared to 7+/-3% (p<0.01) in that pH range in F. In contrast, the predominant isoforms in F eluted in the pH range 8.99-8.0, representing 55+/-8%, while in A the proportion was 11+/-2% (p<0.01). Isoforms eluted at the pH range 7.9-7 represented a significantly greater proportion during A (5.0+/-0.6%) as compared with F (3+/-1%). This is the first report on goat LH circulating isoforms. During A the LH isoforms secreted by the pituitary are more basic than during F.
Assuntos
Anestro/metabolismo , Fase Folicular/metabolismo , Cabras/fisiologia , Hormônio Luteinizante/química , Hormônio Luteinizante/metabolismo , Anestro/efeitos dos fármacos , Animais , Feminino , Fase Folicular/efeitos dos fármacos , Hormônio Liberador de Gonadotropina/farmacologia , Isoformas de Proteínas/química , Isoformas de Proteínas/metabolismoRESUMO
Avaliou-se o papel da gonadotrofina coriônica humana (hCG) e da testosterona na produção de progesterona (P4) e 17ß-estradiol (E2) pelas células da granulosa cultivadas in vitro de folículo antral de égua. Os tratamentos usados foram: 1- controle (nenhum hormônio adicionado), 2- 1UI hCG (0,3µg/ml) e 3- 10UI hCG (3,0µg/ml). O tratamento com hCG foi realizado na presença ou não de testosterona (144ng/ml). O meio foi coletado e substituído com 0,25, 3, 6, 12, 24 e 144h de cultivo. As concentrações de P4 e E2 foram mensuradas por radioimunoensaio. Não se observou diferença entre os tratamentos 1 e 3 quanto à produção de P4 e E2; o tratamento 1 resultou em aumento da concentração de progesterona após 24h de cultura (P<0,01), mas somente em presença de testosterona. A concentração de estradiol aumentou em presença de testosterona, alcançando concentração máxima com 6h de cultura (P<0,01), e diminuiu gradativamente, até atingir a concentração observada com 0,25h de cultura. A adição de hCG não influenciou a síntese do estradiol. A testosterona desempenhou importante efeito estimulador na síntese/secreção doe E2 pelas células da granulosa e modulou a ação do hormônio luteinizante na diferenciação e luteinização das células da granulosa de folículo antral presumidamente pré-ovulatório de égua in vitro.
Assuntos
Animais , Feminino , Gonadotropina Coriônica , Células da Granulosa/metabolismo , Estradiol , Fase Folicular/metabolismo , Cavalos , Técnicas In Vitro , Testosterona/síntese químicaRESUMO
Avaliou-se o papel da gonadotrofina coriônica humana (hCG) e da testosterona na produção de progesterona (P4) e 17ß-estradiol (E2) pelas células da granulosa cultivadas in vitro de folículo antral de égua. Os tratamentos usados foram: 1- controle (nenhum hormônio adicionado), 2- 1UI hCG (0,3µg/ml) e 3- 10UI hCG (3,0µg/ml). O tratamento com hCG foi realizado na presença ou não de testosterona (144ng/ml). O meio foi coletado e substituído com 0,25, 3, 6, 12, 24 e 144h de cultivo. As concentrações de P4 e E2 foram mensuradas por radioimunoensaio. Não se observou diferença entre os tratamentos 1 e 3 quanto à produção de P4 e E2; o tratamento 1 resultou em aumento da concentração de progesterona após 24h de cultura (P<0,01), mas somente em presença de testosterona. A concentração de estradiol aumentou em presença de testosterona, alcançando concentração máxima com 6h de cultura (P<0,01), e diminuiu gradativamente, até atingir a concentração observada com 0,25h de cultura. A adição de hCG não influenciou a síntese do estradiol. A testosterona desempenhou importante efeito estimulador na síntese/secreção doe E2 pelas células da granulosa e modulou a ação do hormônio luteinizante na diferenciação e luteinização das células da granulosa de folículo antral presumidamente pré-ovulatório de égua in vitro.(AU)
The role of the human chorionic gonadotropin (hCG) and testosterone was evaluated in the progesterone (P4) and estradiol-17b (E2) production by granulosa cells of antral follicles from mare cultivated in vitro. The treatment (groups) with gonadotropin consisted of: 1- control (no added hormone); 2- 1 IU hCG (0.3mg/ml) and 3- 10 IU hCG (3.0mg/ml). The treatment with hCG was carried out in the presence or not of testosterone (144ng/ml). The culture medium was collected and replaced at 0.25, 3, 6, 12, 24 and 144h of culture. The concentrations of P4 and E2 were measured by radioimunoassay. Analyses of variance were used for P4 and E2, and mean of the factors were compared by the Tukey test at 5% of probability. No difference was observed between 1 and 2 groups. Treatment with 1 IU of hCG increased progesterone concentration after 24h of culture (P<0.01), only in the presence of testosterone. The concentration of estradiol increased in the presence of testosterone, reaching maximum concentration with 6h of culture (P<0.01), and reduced gradually until the observed concentration at 0.25h of culture. The addition of hCG had no effect in the synthesis of this steroid. The testosterone modulates the action of the luteinizing hormone in the differentiation and luteinization of the granulosa cells and plays important estimulator effect in the synthesis/secretion of E2 by granulosa cells in vitro of presumptive pre-ovulatory antral follicle in mare.(AU)