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The objectives were to develop an effective protocol for transfection of ovine secondary follicles and to assess the effect of attenuating aquaporin 3 (AQP3) using a small interfering RNA (siRNA-AQP3) on antrum formation and follicular growth in vitro. Various combinations of Lipofectamine® volumes (0.5, 0.75 or 1.0 µl), fluorescent oligonucleotide (BLOCK-iT ™) concentrations (3.18, 27.12 or 36.16 nM) and exposure times (12, 14, 16, 18 or 20 h) were tested. The BLOCK-iT™ was replaced by siRNA-AQP3 in the transfection complex. Ovine secondary follicles were isolated and cultured in vitro for 6 days using standard protocols. Follicles were transfected on day 0 or 3 or on both days (0 and 3) and then cultured for an additional 3 or 6 days. As revealed by the fluorescence signal, the Lipofectamine®/BLOCK-iT™ complex (0.75 µl + 27.12 nM by 12 h of incubation) crossed the basement membrane and granulosa cell and reached the oocytes. In general, the rate of intact follicles was higher and the rate of antrum formation was lower in transfected follicles compared with control follicles. In conclusion, ovine secondary follicles can be successfully transfected during in vitro culture, and siRNA-mediated attenuation of AQP3 gene reduced antrum formation of secondary follicles.
The parental bonding is influenced by two dimensions: care and control or protection over the child of both parents. The lack of care during childhood may make the individual more susceptible to the onset of psychiatric disorders when adult. These psychiatric disorders when present during pregnancy may have a negative impact on the health of pregnant women and children. The aim of this study was to assess the association between generalized anxiety disorder (GAD) in pregnant adolescents and the perception of parental bonding.
This is a cross-sectional study with 871 pregnant women under the age of 19, receiving prenatal care in 47 Basic Health Units in the one city, Brazil. The generalized anxiety disorder was measured using the Mini International Neuropsychiatric Interview (MINI) and the perception of parental bonding in childhood using the Parental Bonding Instrument (PBI).
The prevalence of GAD was 8.5%. Among all the parental bonding dimensions, only a perceived lack of maternal care under 16 years was associated with GAD.
The results showed that only the perception of maternal bonding was associated with later GAD. It suggests that an adequate maternal bond is an essential component of psychological health.
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