Effect of prepartum administration of recombinant bovine somatotropin (rbST) on plasma beta-hydroxybutyrate levels in ewes subjected to subclinical ketosis.

O objetivo deste estudo foi determinar o efeito da rbST pré-parto no perfil de plasma BHB de ovelhas gestantes submetidas a cetose subclínica

A HIF-independent, CD133-mediated mechanism of cisplatin resistance in glioblastoma cells

Purpose Glioblastoma Multiforme (GBM) is the commonest brain tumour in adults. A population of cells, known as cancer stem cells (CSCs), is thought to mediate chemo/radiotherapy resistance. CD133 is a cell surface marker to identify and isolate CSCs. However, its functional significance and the relevant microenvironment in which to study CD133 remain unknown. We examined the influence of hypoxia on CD133 expression and the potential functional significance of CD133 in glioblastoma chemoresistance. Methods Gene expression was analysed by qRT-PCR. siRNA technique was used to downregulate genes and confirmed by flow cytometry. IC50 values was evaluated with the Alamar blue assay. Results CD133 expression was upregulated in hypoxia in 2D and 3D models. There was increased resistance to chemotherapeutics, cisplatin, temozolomide and etoposide, in cells cultured in hypoxia compared to normoxia. siRNA knockdown of either HIF1a or HIF2a resulted in reduced CD133 mRNA expression with HIF2a having a more prolonged effect on CD133 expression. HIF2a downregulation sensitized GBM cells to cisplatin to a greater extent than HIF1a but CD133 knockdown had a much more marked effect on cisplatin sensitisation than knockdown of either of the HIFs suggesting a HIF-independent mechanism of cisplatin resistance mediated via CD133. The same mechanism was not involved in temozolomide resistance since downregulation of HIF1a but not HIF2a or CD133 sensitized GBM cells to temozolomide. Conclusion Knowledge of the mechanisms involved in the novel hypoxia-induced CD133-mediated resistance to cisplatin observed might lead to identification of new strategies that enable more effective use of current therapeutic agents

Optimizing biomethane production from anaerobic degradation of Scenedesmus spp. biomass harvested from algae-based swine digestate treatment.

The objective of this work was to quantify biomethane from anaerobic degradation of microalgae biomass harvested from a field-scale tank reactor simulating phycoremediation of swine wastewater. The effects of nutrients starvation on microalgae chemical cellular composition changes and its influence on biomethane generation potential were also addressed. Microalgae polyculture was dominated by uncultured Scenedesmus clone BF 063 which showed a carbohydrate, protein and lipid content of 27.6 ± 3.3, 57.6 ± 0.1 and 3.9 ± 0.6%, respectively. After 25 days exposed to N- and P-free medium, microalgae biomass composition showed 54.6 ± 2.6, 24.1 ± 2.4 and 16.9 ± 0.8% of carbohydrate, protein and lipid, respectively. Volatile solids concentration in the biomass harvested from N- and P-rich medium was lower [67 ± 1.7 g VS (kg biomass)?1] than biomass harvested from nutrient depleted medium [204.1 ± 3.1 g VS (kg biomass)?1]. Consequently, much higher biomethane production was obtained i.e., 103.5 LN CH4 (kg biomass)?1 vs 44 LN CH4 (kg biomass)?1. The results suggest that biomethane production in digesters could be improved by integrating microalgae biomass harvested from algae-based swine wastewater digestate treatment

Cryopreservation of Immature Bovine Oocytes to Reconstruct Artificial Gametes by Germinal Vesicle Transplantation

Joining immature gamete cryopreservation and germinal vesicle transplantation (GVT) technique could greatly improve assisted reproductive technologies in animal breeding and human medicine. The present work was aimed to assess the most suitable cryopreservation protocol between slow freezing and vitrification for immature denuded bovine oocytes, able to preserve both nuclear and cytoplasmic competence after thawing. In addition, the outcome of germinal vesicle transfer procedure and gamete reconstruction was tested on the most effective cryopreservation system. Oocytes, isolated from slaughterhouse ovaries, were stored after cumulus cells removal either by slow freezing or by vitrification in open pulled straws. After thawing, oocytes were matured for 24 h in co-culture with an equal number of just isolated intact cumulus enclosed oocytes, and fixed in order to evaluate the stage of meiotic progression and cytoskeleton organization. Our results showed that after warming, vitrified oocytes reached metaphase II (MII) in a percentage significantly higher than oocytes cryopreserved by slow freezing (76.2% and 36.5% respectively, p < 0.05). Moreover, vitrification process preserved the organization of cytoskeleton elements in a higher proportion of oocytes than slow freezing procedure. Therefore vitrification has been identified as the elective method for denuded immature oocytes banking and it has been applied in the second part of the study. Our results showed that 38.3% of oocytes reconstructed from vitrified gametes reached the MII of meiotic division, with efficiency not different from oocytes reconstructed with fresh gametes. We conclude that vitrification represents a suitable method of GV stage denuded oocyte banking since both nuclear and cytoplasmic components derived from cryopreserved immature oocytes can be utilized for GVT