GSK-3 inhibition reverts mesenchymal transition in primary human corneal endothelial cells

Human corneal endothelial cells are organized in a tight mosaic of hexagonal cells and serve a critical function in maintaining corneal hydration and clear vision. Regeneration of the corneal endothelial tissue is hampered by its poor proliferative capacity, which is partially retrieved in vitro, albeit only for a limited number of passages before the cells undergo mesenchymal transition (EnMT). Although different culture conditions have been proposed in order to delay this process and prolong the number of cell passages, EnMT has still not been fully understood and successfully counteracted. In this perspective, we identified herein a single GSK-3 inhibitor, CHIR99021, able to revert and avoid EnMT in primary human corneal endothelial cells (HCEnCs) from old donors until late passages in vitro (P8), as shown from cell morphology analysis (circularity). In accordance, CHIR99021 reduced expression of α-SMA, an EnMT marker, while restored endothelial markers such as ZO-1, Na+/K+ ATPase and N-cadherin, without increasing cell proliferation. A further analysis on RNA expression confirmed that CHIR99021 induced downregulation of EnMT markers (α-SMA and CD44), upregulation of the proliferation repressor p21 and revealed novel insights into the β-catenin and TGFβ pathways intersections in HCEnCs. The use of CHIR99021 sheds light on the mechanisms involved in EnMT, providing a substantial advantage in maintaining primary HCEnCs in culture until late passages, while preserving the correct morphology and phenotype. Altogether, these results bring crucial advancements towards the improvement of the corneal endothelial cells based therapy

Movable wall and system with environmental condition optimisation functions

The present invention relates to the field of control of indoor environmental conditions. In particular, the invention relates to a movable wall with environmental condition optimisation functions, as well as a system comprising such movable wall, and an environmental condition optimisation method

Nanoneedles Induce Targeted siRNA Silencing of p16 in the Human Corneal Endothelium

Nanoneedles can target nucleic acid transfection to primary cells at tissue interfaces with high efficiency and minimal perturbation. The corneal endothelium is an ideal target for nanoneedle-mediated RNA interference therapy aimed at enhancing its proliferative capacity, necessary for tissue regeneration. This work develops a strategy for siRNA nanoninjection to the human corneal endothelium. Nanoneedles can deliver p16-targeting siRNA to primary human corneal endothelial cells in vitro without toxicity. The nanoinjection of siRNA induces p16 silencing and increases cell proliferation, as monitored by ki67 expression. Furthermore, siRNA nanoinjection targeting the human corneal endothelium is nontoxic ex vivo, and silences p16 in transfected cells. These data indicate that nanoinjection can support targeted RNA interference therapy for the treatment of endothelial corneal dysfunction

Magnesium sulphate in the Emergency Department: an old, new friend

With our study, we searched the medical literature to find magnesium (Mg) correlation with Emergency situations or its use in Emergency Medicine. Our aim is to fill the gap that we find in our daily routine between Mg studies on its role in Emergency and the real conception that doctors have of it in medical practice. We searched the literature for terms as magnesium or magnesium sulphate, magnesium in emergency, eclampsia, arrhythmias, acute asthma exacerbation, magnesium, and pediatric population. After a thorough research, we divided our discoveries into chapters to sort out a large amount often discordant articles

Effects of dietary yeast Saccaromyces cerevisiae on the antioxidant system in the liver of juvenile sea bass Dicentrarchus labrax

The main goal of this work was to determine the effect of dietary live yeast Saccharomyces cerevisiae on the oxidative status of sea bass Dicentrarchus labrax juveniles. Fishes were fed on three diets: the GM group were fed a diet containing lyophilized yeast grown on grape must, the CS group were fed a diet containing lyophilized yeast grown on cornstarch, and the control group were fed a diet without yeast. The activity of the main antioxidative enzymes, superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase, glutathione S-transferase (GST), and glutathione (GSH) content, as well as lipid peroxidation, was measured in the liver of sea bass juveniles 90 days after hatching. Supplementation of the diet with S. cerevisiae significantly reduced the SOD and CAT activity, increased the GST activity, decreased the GSH content, and had no effect on lipid peroxidation. The results support the already reported radical-scavenging properties of yeast and usefulness of its employment as antiperoxidative agent in fish

Potential antioxidant effect of essential oil of Ocinum basilicum in liver of Dicentrarchus labrax

The increasing problems associated with infectious diseases in farmed fish represent major challenges for this source of food production. As the use of antibiotics in aquatic environment poses serious problems at the level of environmental impact and to human health, research on antimicrobial agents from natural sources has received much attention. In addition some naturally occurring antimicrobial agents can be formulated as functional foods and nutraceuticals that can also help to prevent oxidative damage from occurring in the body. The phytochemical preparations with dual functionalities in preventing lipid oxidation and antimicrobial properties have remarkable potential for extending shelf life of food products. As regarding the genus Ocinum, namely Ocinum basilicum, in vitro results are reported on both the antioxidant and antimicrobial activity of its essential oil. In the present communication, an attempt has been made to explore in vivo the possible antioxidant properties of essential oil of O. basilicum in sea bass Dicentrarchus labrax, a widely utilized species in aquaculture, infected with Vibrio alginolyticus. The results obtained show that hepatic lipid peroxidation (measured as TBARS production) induced in D. labrax following infection with V. alginolyticus, is reduced in fish fed with O. basilicum essential oil. In the liver of the same fishes, antioxidant enzymatic activity of glutathione S-transferase (GST) and catalase, is also increased. In light of the results obtained, we suggest that the protective effect of O. basilicum essential oil could be due to activation of the cellular antioxidant system. Further studies are in progress to define the mechanism by which essential oil of O. basilicum could affect the synthesis or regulation of antioxidant enzymes