The action of ethanol on G protein. <i>In silico</i>and cellular/molecular evidences

Ethanol (EtOH) enhances glycinergic currents in the central nervous system (CNS). Because evidence for an interaction between the α1 subunit of the glycine receptor (α1GlyR) and the G protein Gβγ subunit exists in vitro and because cAMP levels are known to increase in response to EtOH, we wanted to investigate the interaction between Gβγ and α1GlyR in response to EtOH treatment in HEK293 cells and to explore the possible sites of interaction between EtOH and the Gαs subunit. His pull-down assays in GlyR-His6-transfected HEK293 cells incubated with ethanol or propofol revealed that only EtOH treatment increased the binding of Gβγ heterodimers to α1GlyR. Using molecular modelling (protein structure prediction), was modelled the hGαs protein for the first time and validated this model by site-directed mutagenesis. By molecular docking, we identified some potential regions of interaction between hGαs and EtOH that are located on the SIII and SI regions of the Gαs. Therefore, we conclude that ethanol increases the interaction between α1GlyR and Gβγ in HEK293 cells, an effect that might be attributed to the interaction between EtOH and hGαs, which consequently stimulates hGαs.

Microencapsulation with spray-chilling as an innovative strategy for probiotic low sodium requeijao cremoso processed cheese processing

The effect of sodium reduction and Lactobacillus acidophilus addition(free or microencapsulated)on the quality parameters of requeijao cremoso processed cheese was assessed for 90 days at 5℃.Three formulations were prepared:(CONT)regular salt content and curd fusion(90℃/2 min),(FREE)reduced salt content+free probiotic culture added after curd fusion(90℃/2 min),and(MICRO)reduced salt content+microencapsulated probiotic culture added before curd fusion(70℃/5 min).FREE and MICRO formulations showed increased monounsaturated and polyunsaturated fatty acid content,improved health indices(decreases in thrombogenic index and increases in desirable fatty acid index),and altered rheological properties(higher consistency index,apparent viscosity,elastic properties,and gel strength).In addition,the MICRO formulation showed higher texture sensory acceptance and probiotic counts higher than 6 log CFU/g during storage and simulated gastrointestinal conditions.Overall,microencapsulation of probiotics with spray chilling represents an innovative solution in requeijao cremoso processed cheese.