[Prospects for the correction of intestinal microbiota in the prevention and treatment of asthma in children].

The issues of therapy and prevention of asthma in children do not lose their relevance. The increase in the prevalence of allergic diseases and asthma is associated, among other things, with a relative deficit in the microbial load and changes in the microbiota due to improved hygiene and living conditions. The microbiota plays an important role in the formation and functioning of the immune system. Contact with microorganisms contributes to the normal maturation of T-regulatory cells, preventing an inadequate immune response in both the Th1- and Th2-pathways. Aim - literature review on the possibilities of gut microbiota correction for prevention and treatment of asthma in children. Results. In children with an increased risk of asthma, abnormalities in the intestinal microbiota are observed in the first year of life: there is a relative deficiency of Lachnospira, Veillonella, Faecalibacterium and Rothia. The imbalance of the intestinal microbiota is accompanied by a decrease in the synthesis of short-chain fatty acids (butyrate, acetate, propionate), which, among other things, play the role of signaling molecules. Gut microbiota maturation is delayed in children at risk of asthma. It is not clear whether the diversity of the gut microbiota is associated with the risk of developing asthma. Maintenance of normal gut microbiota or correction of its disturbances in early life is a possible approach to the prevention and treatment of asthma. Breastfeeding, vaginal delivery, constant contact with farm animals or dogs since an early age, limiting antibiotic use in the first year of life, a varied diet with the inclusion of fiber-rich foods, and the use of pre- and probiotics can help. However, the implementation of these recommendations in practice is difficult. Further research is required to identify specific prophylactic stimuli reproducible in the urban environment. The conflicting results of studies in this area, in particular, the effectiveness of probiotics in the prevention of asthma, require large-scale prospective cohort studies with a long follow-up period and careful selection of probiotic strains and their combinations. Now, there are no definitive recommendations on the use of probiotics for the prevention of allergic diseases. Conclusion. The possibilities of correcting the gut microbiota for the prevention and treatment of asthma are actively studied, but at present, there are many contradictions and unresolved issues.

Proteomic analysis of nuclei dissected from fixed rat brain tissue using expression microdissection.

Expression microdissection (xMD) is a high-throughput, operator-independent technology that enables the procurement of specific cell populations from tissue specimens. In this method, histological sections are first stained for cellular markers via either chemical or immuno-guided methods, placed in close contact with an ethylene vinyl acetate (EVA) film, and exposed to a light source. The focal, transient heating of the stained cells or subcellular structures melts the EVA film selectively to the targets for procurement. In this report, we introduce a custom-designed flashcube system that permits consistent and reproducible microdissection of nuclei across an FFPE rat brain tissue section in milliseconds. In addition, we present a method to efficiently recover and combine captured proteins from multiple xMD films. Both light and scanning electron microscopy demonstrated captured nuclear structures. Shotgun proteomic analysis of the samples showed a significant enrichment in nuclear localized proteins, with an average 25% of recovered proteins localized to the nucleus, versus 15% for whole tissue controls (p < 0.001). Targeted mass spectrometry using multiple reaction monitoring (MRM) showed more impressive data, with a 3-fold enrichment in histones, and a concurrent depletion of proteins localized to the cytoplasm, cytoskeleton, and mitochondria. These data demonstrate that the flashcube-xMD technology is applicable to the proteomic study of a broad range of targets in molecular pathology.