Influence of L-arginine during bovine in vitro fertilization.

The objective of this work was to evaluate the effect of using L-arginine during in vitro fertilization (IVF) on in vitro embryonic development using Bos taurus and Bos indicus semen. Effect of different concentrations (0, 1, 10 and 50 mM) of L-arginine, added to the IVF medium, was evaluated on the fertilization rate at 18 h post-fertilization (hpf), NO3-/NO2- production during IVF by the Griess colorimetric method (30 hpf), cleavage and blastocyst rates (on Day 2 and Day 7 of culture, respectively) and total blastocyst cell number (Day 7 of culture). The results reveal that the addition of 50 mM L-arginine to IVF medium, with either Bos taurus or Bos indicus spermatozoa, decreased the cleavage rate and blastocyst rate compared to the control group. Other concentrations did not affect embryo production. However, 1 mM L-arginine with Bos indicus semen increased the proportion of hatched blastocysts. These results indicate that high L-arginine concentrations may exhibit toxic effects on bovine gametes during in vitro fertilization.

Os efeitos do desidroepiandrosterona na resposta ovariana de más respondedoras submetidas à reprodução humana assistida / Effects of dehydroepiandrosterone on ovarian response of poor responders undergoing assisted human reproduction

Os androgênios, por agirem de forma positiva no desenvolvimento folicular, estão sendo atualmente utilizados na reprodução humana assistida como uma alternativa para melhorar a resposta ovariana de mulheres consideradas más respondedoras. Esta revisão sistemática avalia o efeito do desidroepiandrosterona (DHEA) na resposta à estimulação ovariana de mulheres más respondedoras submetidas às técnicas de reprodução assistida. Os artigos para este estudo foram pesquisados no PubMed e publicados entre 1999 e 2013. Vinte e sete artigos foram avaliados e 18 deles foram selecionados, incluindo estudos experimentais e observacionais. O DHEA foi associado a um maior número de folículos recrutados, de oócitos selecionados e melhor qualidade embrionária, à diminuição do risco de aneuploidias e à maior taxa de gravidez clínica e nascidos vivos. Apesar de o DHEA apresentar efeito positivo na resposta ovariana de mulheres más respondedoras, os resultados obtidos foram pouco consistentes. Mais estudos controlados e randomizados devem ser realizados antes de se implantar o DHEA de rotina no tratamento de más respondedoras submetidas à reprodução humana assistida.(AU)

Androgens are currently being used in assisted human reproduction as an alternative to improve ovarian response of women considered poor responder by acting positively in follicular development. This systematic review evaluates the effects of dehydroepiandrosterone (DHEA) in response to ovarian stimulation of poor responder women undergoing assisted reproductive techniques. All articles for this study were searched in PubMed and published between 1999 and 2013. Twenty seven articles were evaluated and 18 of them were selected, including experimental and observational studies. DHEA was associated with a greater number of follicles, oocyte selected and better embryo quality, the decreased risk of aneuploidy and higher rates of clinical pregnancy and live birth. Although DHEA has positive effect on the ovarian response of poor responder women, the results were inconsistent. More randomized controlled trials should be conducted before using DHEA in routine treatment of poor responders undergoing assisted reproduction.(AU)

effect of retinoic acid on in vitro maturation and fertilization rate of mouse germinal vesicle stage oocytes

Retinoids are recognized as important regulators of cell differention and tissue function, Previous studies, performed both in vivo and in vitro, indicate that retin-oids influence several reproductive events. In this study, we investigated the effect of all-trans retinoic acid [t-RA] on maturation and fertilization rate of immature oocytes [germinal vesicle]. Germinal vesicle [GV] oocytes were recovered from 4-6 week old female mice 48 hours after injection of 10 IU pregnant mare serum gonadotropin [PMSG]. Collected oocytes were divided into seven groups: control, sham and five experimental groups. t-RA at concentrations of 1, 2, 4, 6, 8 jjM were added to oocyte maturation medium in the experimental groups. The maturation rate was recorded after 24 hours of culture in a humidified atmosphere of 5% CO[2] at 37°C. Fertilization and developmental rates of matured oocytes were recorded after in vitro fertilization [IVF] and 24 hour culture. The rate of oocytes that developed to the metaphase II stage of maturation significantly increased with 2 and 4 microM t-RA compared to the control and sham groups [p<0.05]. In addition, the number of fertilized oocytes was significantly higher in 4 microM retinoic acid compared to the control [p<0.05], but the difference between the number of fertilized oocytes which developed to the 2-cell stage was not significant between the two groups. The results show that t-RA enhanced mouse oocyte maturation in vitro and improved fertilization and development rates in a dose dependent manner

[Effect of demecolicne treatment on the developmental competency of in vitro matured bovine oocytes]

The aim of this study was to investigate whether demecolicne treatment of matured bovine oocytes adversely affects the process of in vitro fertilization and embryo development. Bovine Cumulus Oocyte Complexes [COC's] were matured in vitro and then were randomly allocated to two treatment groups of common concentrations of demecolicne [0.05 and 0.4 micro g/ml for 30 min] and a control group. COC's were then fertilized and cultured in vitro for up to 9 days when the ratios of in vitro embryo development and the viability of the hatched blastocysts were assessed and compared with the control group [p<0.05]. The ratios of the cleavage and blastocyst formation of demecolicne treated groups [0.4 and 0.05 micro g/ml] were 68.6, 63.5% and 23.3, 32.8%, which were not significantly different from the control group [73.3, 29.0%], respectively. The results of cell-viability were also not significantly different between the control vs. treatment groups. Since the overall indices of in vitro embryo development revealed no significant difference between the demecolicne treated compared to control bovine oocytes, it seems that demecolicne treatment of matured bovine oocytes may not compromise their potency for further in vitro development

Effect of supplementation of green tea polyphenols on the developmental competence of bovine oocytes in vitro

The objective of the present study was to examine the effect of green tea polyphenols (GTPs) supplementation during in vitro maturation, in vitro fertilization, and in vitro culture on the developmental competence of bovine oocytes. Cumulus-oocyte complexes aspirated from the ovaries were matured in vitro (38.5°C for 24 h) and fertilized (38.5°C for 15-18 h) and embryos were cultured (38.5°C for 192 h) in a defined conditioned medium with or without GTPs supplementation. The GTPs used in the present study contained 99 percent catechin derivatives, with the major components being 50 percent (-)-epigallocatechin gallate, 22 percent (-)-epicatechin gallate, 18 percent (-)-epigallocatechin, and 10 percent (-)-epicatechin. Four replicate trials were done for each type of experiment. GTPs supplementation (15 æM) of the maturation medium led to a significant increase in the rate of blastocyst formation (34.0 vs 21.4 percent, P < 0.05). However, the rate of blastocyst formation was not improved when higher GTPs concentrations (20 or 25 æM) were added to the in vitro maturation medium. During in vitro fertilization, supplementation with higher GTPs concentrations (20 or 25 æM) significantly reduced the rate of blastocyst formation (P < 0.05). Supplementation of the culture medium with 15 æM GTPs improved the rate of blastocyst formation, while higher GTPs concentrations (25 æM) significantly reduced embryo development (P < 0.05). In conclusion, these results demonstrate that supplementation with GTPs at low concentration (15 æM) during in vitro maturation and in vitro culture improved the developmental competence of bovine oocytes.

Ketorolac versus fentanyl for analgesia during vaginal ovum pickup procedures

Thirty adult females ASA class I or II undergoing vaginal ovum pickup procedures, were randomly allocated to receive either i.v. fentanyl [100 ug], ketorolac [60 mg] or both fentanyl [50 ug] and ketorolac [30 ug] 30 min before the procedure. Hemodynamic variables showed significant rise in MAP, HR and RR during the procedure in ketorolac group, while there were no significant changes in both fentanyl and the combination groups. But postoperatively no significant changes were detected. Postoperatively, comfort score was higher [3.7] in ketorolac group compared to fentanyl group [2.5] and combination group [2.3]. Also, VAS scores were higher in ketorolac group [37] compared to fentanyl group [22] and combination group [33]. The incidence of nausea, vomiting, or both, and the incidence of antiemetic therapy were higher in fentanyl group only. So, ketorolac [60 mg] appears to have less analgesia and comfort with less side effects than fentanyl [100 mug]. But combination of ketorolac [30 mg] with fentanyl [50 mug] appears to be better than each of them alone