A Method for Analysis of Nitrotyrosine-Containing Proteins by Immunoblotting Coupled with Mass Spectrometry.

Nitrotyrosine formation is caused by presence of reactive oxygen and nitrogen species. Nitration is a very selective process leading to specific modification of only a few tyrosines in protein molecule. 2D electrophoresis and western blotting techniques coupled with mass spectrometry are common methods used in analysis of proteome. Here we describe protocol for analysis of peroxynitrite-induced protein nitration in isolated mitochondria. Mitochondrial proteins are separated by 2D electrophoresis and transferred to nitrocellulose membrane. Membranes are then incubated with antibodies against nitrotyrosine. Positive spots are compared with corresponding Coomassie-stained gels, and protein nitration is confirmed with mass spectrometry techniques.

Low-carbohydrate diet in diabetes mellitus treatment.

There has been an increasing amount of information about the positive results of low-carbohydrate diet in the treatment of diabetes, pre-diabetes, metabolic syndrome and obesity in the form of randomized trials, their meta-analysis and case studies. Many of these indicate that low carbohydrate diets are safe, could significantly improve the compensation of both types of diabetes and the overall health of the diabetic patients. In successful therapy, this diet leads to weight loss, lower medication doses or prescribing, and in some cases of type 2 diabetes also to remission. However, the low carbohydrate diet is not recognized in Czech diabetology, and concerns remain particularly about its safety. This article is a summary of the current knowledge about low-carbohydrate diet, its benefits, risks and contraindications, and aims to initiate a discussion about its use as one of the options for dietary treatment of diabetics. Key words: diabetes mellitus - ketogenic diet - low-carbohydrate diet - metabolic syndrome - obesity.