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Objective: To investigate if the protective effect of α-tocopherol against the impact of ethanol on brain morphogenesis involved the activity of the selenoproteins phospholipid hydroperoxide glutathione peroxidase (PHGPx; GPx4) and selenoprotein P (SelPP) that have roles against oxidative stress.Methods: Forty female mice were randomly assigned into natural control (CON), positive control (ETOH), low-, medium-, and high-α-tocopherol-supplemented-ethanol groups (LTOC, MTOC, HTOC, respectively). CON received drinking water without ethanol while ETOH, LTOC, MTOC and HTOC groups received 20% ethanol in drinking water. The supplemented groups were given respective dosages of α-tocopherol, 0.410, 0.819, and 1.640 mg/g body weight, at day 14 before mating onwards to the day 9 of gestation. At 10.5 ED of gestation (1 100 h), the pregnant females were sacrificed by cervical dislocation and the embryos were harvested. Total RNA were extracted, cDNA synthesis and qRT-PCR analyses were carried out.Results: The level of expression of PHGPx in the positive control was significantly lower than that of the natural control. Among the three α-tocopherol-supplemented groups, only the medium dose-group was significantly higher than the positive control. The level of expression of SelPP in the positive control was significantly lower than those of the natural control, the low- and medium-dose-tocopherol supplemented groups. In the high dose-α-tocopherol supplemented group, the level of expression was not significantly different from the positive control but significantly lower than the natural control.Conclusions: The activity of the selenoproteins PHGPx and SelPP are involved in the internetwork of antioxidative enzymes with vitamin E when given up to a medium dose only and is one of the possible pathways of shielding embryonic development against the impact of ethanol on brain morphogenesis. This study strengthens the impact of dietary α-tocopherol and Selenium supplement during the critical period of pregnancy.

Objective To assess the histoarchitecture of the maternal liver and kidney as influenced by selenium supplementation within the periconception period and to determine which of the periconception stages of selenium intake is most favorable of the liver and kidney. Methods Thirty-six 7-week-old female mice were divided into four groups where all groups were given 6.0 g of food pellets per day. Un-supplemented group (U) received no selenium in the food pellet, Pre-gestation supplemented group (P) was given 3.0 μg selenium/d for 21 d, Pre-gestation to Gestation supplemented group (PG) was given 3.0 μg selenium/d for 37 d, and Gestation supplemented group (G) was given 3.0 μg selenium/d for 16 d only. Results The occurrences of vacuolated hepatocytes and congested central veins in the liver were observed. The P and the G groups incurred the lowest and highest percent occurrence, respectively, of murine mothers that exhibited presence of vacuolated hepatocytes. The mean percent occurrence of congested central veins did not significantly vary among groups. The occurrences of shrunken and swollen glomeruli were observed in the kidney. The mean percent occurrence of shrunken glomeruli showed that P and G groups were significantly lower than those of U and PG groups. The mean percent occurrences of swollen glomeruli did not vary significantly among groups. Conclusion The histological analyses of the liver and the kidney obtained from different stages of periconception indicate that selenium supplementation would be best during pregestation stage.

Objective: To assess the protective potential of α-tocopherol on ethanol-induced dysmorphogenesis in 10.5 embryonic day (ED) mouse embryos. Methods: Forty female mice were randomly assigned into control (CON), positive control (ETOH), low-, medium and high-α-tocopherol-supplemented-Ethanol groups (LTOC, MTOC, HTOC, respectively). CON received drinking water without ethanol, ETOH, LTOC, MTOC and HTOC groups received 20% ethanol in drinking water. The supplemented groups were given respective dosages of α-tocopherol, 0.410, 0.819 and 1.640 mg/g body weight, at 14 days before mating until the 9th day of gestation. The 10.5 ED embryos were assessed for embryo weight, head- and crown-rump length, and morphological scoring of brain and sensory vesicles, flexion and somites. The embryo yield was assessed by counting the number of full-term developed embryos from the bulging implantation sites while resorption was assessed by counting the bulging implantation sites but without formed embryos. Results: The weight and head- and crown-rump length of the embryos from the α-tocopherol supplemented groups were comparable to the control. These were significantly higher than that of positive control (P

Objective To determine the influence of maternal Isoflavone-supplementation during peri-conception period on the reproductive organs of the first generation (F1) murine weanling-stage offspring. Methods Sexually-mature female mice were randomly distributed into the Control group (C) that was fed with food pellets only, treatment groups that were designated as High Dose (HD), Medium Dose (MD) and Low dose (LD) that were fed with food pellets + Isoflavone Genistein dosages of 150.0, 100.0, and 50.0 mg/kg body weight, respectively. Supplement was given two weeks prior to mating and throughout a 21-day gestation period. The pups that were delivered on the 21st day were allowed to mature up to the 28th-day postnatal age then sacrificed by cervical dislocation. The testis and epididymis of males and ovaries and oviducts of females were obtained for histological analyses. Results Histological analysis of the testes showed clogged luminal spaces and indistinct population of spermatogenic cells in the HD group while the development of the ovaries from all test groups appeared intact. The luminal and tubular diameter of epididymis were unaffected in the isoflavone-supplemented groups. The epididymis of HD group exhibited poorly developed cauda segment and atrophy of the caput segment but not for MD and LD groups. The oviducts of the HD and MD groups exhibited collapsed muscularis layer and indistinguishable mucosa layer with HD being the more severe. The LD and Control groups showed well-developed mucosa layer. Conclusion Results showed that maternal isoflavone-supplementation when administered at a high dose of 150 mg/kg body weight during peri-conception period can induce adverse effects on the reproductive organs of first generation (F1) murine weanling-stage offspring.

Objective To investigate the effects of pregestational and gestational exposure to an isoflavone of sexually mature female mice on their reproductive health and on the postnatal development of their offspring. Methods Sexually-mature ICR female mice were randomly segregated into Control group 1 that was given the normal diet consisting of food pellets, Control group 2 that was given food pellets with additional corn oil of 2 mL/kg bodyweight. The three (3) isoflavone treatment groups were given 50 mg/kg body weight, 100 mg/kg body weight and 150 mg/kg body weight for low dose (LD), medium dose (MD) and high dose (HD) respectively. Treatments were administered 2 weeks prior to mating and gestation and thereafter until parturition. The delivered pups were weaned up until 21 days. On the 21st day, postnatal development were determined, excluding the birth weight of pups which was measured one day post-parturition. The dams were sacrificed as well and the markers of maternal health were determined. Results There were no significant differences found between the control groups and the treatment groups in terms of the markers of maternal reproductive health. For postnatal development, only HD group displayed a significantly higher mean AGD from the other groups. Conclusions The data implies that exposure to the isoflavone genestein, with the given dosages, does not impact the maternal reproductive health while the high dose brings about masculinization of the pups which implies that isoflavone exerts its action as an endocrine disruptor affecting postnatal development. This could be attributed to the decrease in estrogen due to the inhibition of aromatase, an enzyme involved in estrogen synthesis.

Cdc6 is a key regulator of DNA replication in eukaryotes. In this work, the expression pattern of an Arabidopsis cdc6 homologue is characterized by RT-PCR and in situ hybridization. The data suggest that cdc6At expression is cell cycle regulated. During development, high cdc6At mRNA levels are found in regular cycling cells. In addition, cdc6At expression is also observed in cells that are probably undergoing endoreduplication, suggesting a possible role of Cdc6At in this process in plants.