RESUMEN

This study evaluated the association of polymorphisms of VEGF (endothelial vascular growth factor) gene + 936C/T (rs3025039), 1154 G/A (rs 1570360) and -2578 C/A (rs 699947) in patients with polycystic ovary syndrome (PCOS) and to perform the haplotypes formed by the alleles in the Brazilian population. A total of 110 women without PCOS and 112 women with PCOS were included in the study. Genotyping analyses were performed using the PCR-RFLP assays (rs 3025039 and rs 699947) and by allelic discrimination using the real-time PCR technique (rs 1570360). In the univariate analysis, we observed a significant difference between the groups for the polymorphism rs 1570360 and this polymorphism presented statistical differences between the groups for the recessive model (p = .04). The frequency of the T-G-C haplotype showed a statistically significant difference between women with PCOS and controls (p = .05). The -2578 A/C polymorphism was more frequent in the control group, which may be associated with a protective characteristic for the PCOS manifestation. In the sample analysis, polymorphism rs 1570360 is associated with PCOS and the T-G-C haplotype could be associated with protective factors.

Asunto(s)

RESUMEN

RESEARCH QUESTION: Does a three-dimensional culture system based on magnetic levitation with nanoparticles assembly maintain the follicular structure and viability with adequate growth rates leading to oocyte maturation after long-term culture? DESIGN: Randomized-controlled trial of treatments in a bovine model. Secondary follicles (n = 213) isolated from bovine ovaries were cultured in a two-dimensional system (two-dimensional control) or three-dimensional levitation system with different concentrations (three-dimensional 50 µl/ml, 100 µl/ml and 200 µl/ml) of magnetic nanoparticles. Follicular growth (diameter, daily growth and growth patterns), morphology (normal, degenerated and extruded follicles), antrum formation, oocyte viability and chromatin configuration were assessed. RESULTS: Secondary follicles of three-dimensional 200-µl/ml treatment showed higher viability, antrum formation and lower degeneration rates than two-dimensional control. Also, follicles cultured in the three-dimensional 200-µl/ml treatment presented a most homogenous daily growth rate as shown by the lowest variance and standard deviation. Compared with the two-dimensional control, the proportion of non-growing and slow-growing follicles were 3.8-fold lower and 1.6-fold higher, respectively, in the three-dimensional 200-µl/ml treatment. After in-vitro maturation, the three-dimensional 200-µl/ml had a greater proportion of viable oocytes (1.7-fold) and meiotic resumption rates (2.4-fold) than the two-dimensional control treatment. CONCLUSION: The three-dimensional levitation culture system improves the viability of in-vitro development of bovine secondary follicles, antrum formation and lower extrusion and degeneration rates and adequate growth rate leading to relevant oocyte viability and meiotic resumption after in-vitro maturation. This approach does not require a specific medium, and has the potential as an alternative method to in-vitro follicle culture in several species, including humans.

Asunto(s)

RESUMEN

The aim of this study was to investigate the effect of Kisspeptin (Kp) on the medium used in different stages of in vitro production of bovine embryo (IVEP), evaluating cleavage (CR) and blastocyst (BR) rates. The study was divided into three experiments that analyzed, respectively, the action of Kp on in vitro maturation (IVM), in vitro fertilization (IVF), and in vitro culture (IVC) of bovine embryos. In experiment 1, the oocytes were matured in IVM medium and distributed into the following treatments: maturation (IVM Control, n = 102), maturation with addition of 10-7 M Kp (Kp 10-7 IVM, n = 90), and hormone-free maturation luteinizing hormone (LH) and follicle-stimulating hormone (FSH) with the addition of 10-7M Kp (No hormones + Kp 10-7, n = 84), following maturation to normal stages of IVEP. In experiment 2, the oocytes were fertilized in IVF medium, in the following treatments: TALP-FERT without Kp (Control IVF, n = 103) and TALP-FERT with the addition of 10-7M Kp (Kp 10-7 IVF, n = 119), usually following the other steps. Finally, in the third experiment, the oocytes passed through all phases and were divided into IVC in two treatments: SOF medium without Kp (Control IVC, n = 109) and SOF medium with the addition of 10-7M Kp (Kp 10-7, N = 106). The data were analyzed by PROC GLIMMIX of the SAS program. In experiment 1, the means of CR and BR were similar (P > 0.05) between treatments (IVM Control76.47% and 37.25%, Kp 10-7 MIV80% and 33.33%, and No hormones + Kp 10-770.24% and 30.95%, respectively). In experiment 2, the means of CR were similar for the IVF Control and Kp 10-7 IVF groups (P > 0.05), 76.70% and 86.55% respectively. But, the mean of the BR of the group Kp 10-7 IVF was 38.66%, which was higher (P 0.05) were similar between the IVC Control and Kp 10-7 IVC groups (CR 83.50% and 78.30%, and BR 26.60% and 23.60%, respectively)...

Objetivou-se com esse estudoinvestigar o efeito da Kisspeptina (Kp) nos meios base utilizados nas etapas da Produção in vitro de Embriões (PIVE) bovinos, avaliando as taxas de clivagem (TC) e blastocistos (TB). O estudo foi dividido em três experimentos, sendo respectivamente analisado em cada um a ação da Kp na maturação in vitro (MIV), fertilização in vitro (FIV) e cultivo in vitro (CIV) de embriões bovinos. No experimento 1 os oócitos foram maturados em meio base MIV, distribuídos nos seguintes tratamentos: maturação (Controle MIV, n=102), maturação com adição de 10-7M de Kp-10 (Kp 10-7 MIV, n=90) e maturação sem hormônios LH e FSH com adição de 10-7M de Kp-10 (Sem hormônios + Kp 10-7, n=84), seguindo após a maturação para as etapas normais da PIVE. No experimento 2 os oócitos foram fecundados em meio base da FIV, nos seguintes tratamentos:TALP-FERT sem Kp (Controle FIV, n = 103) e TALP-FERT com adição de 10-7M de Kp-10 (Kp 10-7 FIV, n = 119), seguindo normalmente pelas outras etapas. Finalmente, no terceiro experimento os oócitos passaram por todas as fases e foram divididos no CIV em 2 tratamentos: meio SOF sem Kp (Controle CIV, n = 109) e meio SOF com adição de 10-7M de Kp-10 (Kp 10-7 3, n = 106). Os dados foram analisados pelo PROC GLIMMIX do programa SAS. No experimento 1 as médias das TC e TB foram semelhantes (P > 0,05) entre os tratamentos Controle MIV 76,47% e 37,25%, Kp 10-7 MIV 80% e 33,33% e Sem hormônios + Kp 10- 7 70,24% e 30,95%, respectivamente. No experimento 2 as médias das TC foram semelhantes para os grupos Controle FIV e Kp 10-7 FIV (P > 0,05), sendo 76,70% e 86,55%, respectivamente. Porém, a média da TB do grupo Kp 10-7 FIV 38,66%, foi superior (P 0,05) também foram similares entre os tratamentos Controle CIV e Kp 10-7 CIV...

Asunto(s)

RESUMEN

The aim of this study was to investigate the effect of Kisspeptin (Kp) on the medium used in different stages of in vitro production of bovine embryo (IVEP), evaluating cleavage (CR) and blastocyst (BR) rates. The study was divided into three experiments that analyzed, respectively, the action of Kp on in vitro maturation (IVM), in vitro fertilization (IVF), and in vitro culture (IVC) of bovine embryos. In experiment 1, the oocytes were matured in IVM medium and distributed into the following treatments: maturation (IVM Control, n = 102), maturation with addition of 10-7 M Kp (Kp 10-7 IVM, n = 90), and hormone-free maturation luteinizing hormone (LH) and follicle-stimulating hormone (FSH) with the addition of 10-7M Kp (No hormones + Kp 10-7, n = 84), following maturation to normal stages of IVEP. In experiment 2, the oocytes were fertilized in IVF medium, in the following treatments: TALP-FERT without Kp (Control IVF, n = 103) and TALP-FERT with the addition of 10-7M Kp (Kp 10-7 IVF, n = 119), usually following the other steps. Finally, in the third experiment, the oocytes passed through all phases and were divided into IVC in two treatments: SOF medium without Kp (Control IVC, n = 109) and SOF medium with the addition of 10-7M Kp (Kp 10-7, N = 106). The data were analyzed by PROC GLIMMIX of the SAS program. In experiment 1, the means of CR and BR were similar (P > 0.05) between treatments (IVM Control76.47% and 37.25%, Kp 10-7 MIV80% and 33.33%, and No hormones + Kp 10-770.24% and 30.95%, respectively). In experiment 2, the means of CR were similar for the IVF Control and Kp 10-7 IVF groups (P > 0.05), 76.70% and 86.55% respectively. But, the mean of the BR of the group Kp 10-7 IVF was 38.66%, which was higher (P < 0.05) than that of the FIV Control group, which was 31.07%. In the third experiment, the means of CR and BR (P > 0.05) were similar between the IVC Control and Kp 10-7 IVC groups (CR 83.50% and 78.30%, and BR 26.60% and 23.60%, respectively)...(AU)

Objetivou-se com esse estudoinvestigar o efeito da Kisspeptina (Kp) nos meios base utilizados nas etapas da Produção in vitro de Embriões (PIVE) bovinos, avaliando as taxas de clivagem (TC) e blastocistos (TB). O estudo foi dividido em três experimentos, sendo respectivamente analisado em cada um a ação da Kp na maturação in vitro (MIV), fertilização in vitro (FIV) e cultivo in vitro (CIV) de embriões bovinos. No experimento 1 os oócitos foram maturados em meio base MIV, distribuídos nos seguintes tratamentos: maturação (Controle MIV, n=102), maturação com adição de 10-7M de Kp-10 (Kp 10-7 MIV, n=90) e maturação sem hormônios LH e FSH com adição de 10-7M de Kp-10 (Sem hormônios + Kp 10-7, n=84), seguindo após a maturação para as etapas normais da PIVE. No experimento 2 os oócitos foram fecundados em meio base da FIV, nos seguintes tratamentos:TALP-FERT sem Kp (Controle FIV, n = 103) e TALP-FERT com adição de 10-7M de Kp-10 (Kp 10-7 FIV, n = 119), seguindo normalmente pelas outras etapas. Finalmente, no terceiro experimento os oócitos passaram por todas as fases e foram divididos no CIV em 2 tratamentos: meio SOF sem Kp (Controle CIV, n = 109) e meio SOF com adição de 10-7M de Kp-10 (Kp 10-7 3, n = 106). Os dados foram analisados pelo PROC GLIMMIX do programa SAS. No experimento 1 as médias das TC e TB foram semelhantes (P > 0,05) entre os tratamentos Controle MIV 76,47% e 37,25%, Kp 10-7 MIV 80% e 33,33% e Sem hormônios + Kp 10- 7 70,24% e 30,95%, respectivamente. No experimento 2 as médias das TC foram semelhantes para os grupos Controle FIV e Kp 10-7 FIV (P > 0,05), sendo 76,70% e 86,55%, respectivamente. Porém, a média da TB do grupo Kp 10-7 FIV 38,66%, foi superior (P < 0,05) ao grupo controle FIV 31,07%. No terceiro experimento as médias das TC e TB (P > 0,05) também foram similares entre os tratamentos Controle CIV e Kp 10-7 CIV...(AU)

Asunto(s)