[The influence of environmental factors on the prevalence of «thrifty genotypes¼ as predictors of metabolic disorders].

"Thrifty genotypes" are the risk factors for obesity and lipid and energy metabolism disorders. Hence, it is important to assess the contribution of environmental factors that influenced the thrifty genotypes' population distribution. Aim of the study - systematization and critical analysis of published data on population variability, relationship with climatic and environmental characteristics, association with traditional types of lifestyles, and nutrition for the «thrifty genotypes¼ of APOE, UCP1, UCP3, and FTO genes. Material and methods. The selection of publications from the last 20-25 years presented in the PubMed database (https://pubmed.ncbi.nlm.nih.gov) was carried out by the keywords of the generalizing rank (thrifty genotype, thrifty phenotype, drifty genotype), then narrowed down to the APOE, UCP, FTO. The final set includes publications that consider the association of genotypes with the ecological conditions of the population. Results. Our analysis of publications has confirmed the ethnic and geographical variability in the allele distribution of APOE, UCP1, UCP3, and FTO genes. However, the nature of this variability hasn't been studied sufficiently; the contribution of individual factors of the natural and anthropogenic environment remains unclear. The information on the geographical distribution of the APOE gene alleles is quite complete, while the data on the «thrifty genotypes¼ of UCP and FTO require further study. Conclusion. The frequency of the UCP1 and UCP3 alleles associated with effective non-contractile thermogenesis is increased in populations adapted to low temperatures. However, the population-geographical pattern of the UCP thrifty genotypes' variability as a determinant of increased fat deposition has been studied insufficiently. The carriage of FTO mutant variants increases the adaptability of groups with a traditional lifestyle and diet but is maladaptive in an urbanized environment. The influence of natural and ecological conditions on the formation of the FTO allele geographical distribution requires more attention. The results obtained allow us to propose the included groups' ranking according to the past environmental management and nutrition will facilitate the search for ecological factors that influenced the geographical distribution of genotypes (and, accordingly, populations with different levels of risk of metabolic disorders).

Brain derived neutrophic factor, a link of aerobic metabolism to neuroplasticity.

Currently, literature has accumulated great knowledge over the effect of exercise on the neurotrophin named brain derived neurotrophic factor (BDNF) and its role in neuronal plasticity. However, there is no enough discussion about how the exercise is related to enrichment of BDNF in specific metabolic properties. This review provides the current evidences regarding aerobic metabolism relation to BDNF concentrations in healthy individuals. A PICOS strategy was applied considering the mesh terms for: P - healthy subjects; I - physical exercise; C - aerobic metabolism demands; O - BDNF concentrations; S - before and after aerobic exercise; on PubMed, Scopus and Medline databases. Studies presenting at least one session the exercise with reports of BDNF analysis before and after were included. Reviews, letters, case-reports, articles not written in English, non- published or involving non-healthy populations were excluded. Compiling results, it was possible to observe a close interaction between different aerobic energy demands from the exercise models and the responses of BDNF, suggesting thus that increases in BDNF concentrations are associated to the amount of aerobic energy required by exercise in a dose-dependent manner. Moreover, the dynamics of BDNF synthesis and reuptake resemble the functioning of the metabolic systems of aerobic energy generation, with which they share a co-transcriptional factor dependence.

[Solid state isotope hydrogen exchange for deuterium and tritium in human gene-engineered insulin].

The reaction of high temperature solid state catalytic isotope exchange in peptides and proteins under the action of catalyst-activated spillover hydrogen was studied. The reaction of human gene-engineered insulin with deuterium and tritium was conducted at 120-140° C to produce insulin samples containing 2-6 hydrogen isotope atoms. To determine the distribution of the isotope label over tritium-labeled insulin's amino acid residues, oxidation of the S-S bonds of insulin by performic acid was performed and polypeptide chains isolated; then their acid hydrolysis, amino acid analysis and liquid scintillation counts of tritium in the amino acids were conducted. The isotope label was shown to be incorporated in all amino acids of the protein, with the peptide fragment FVNQHLCGSHLVE of the insulin ß-chain showing the largest incorporation. About 45% of the total protein isotope label was incorporated in His5 and His10 of this fragment. For the analysis of isotope label distribution in labeled insulin's peptide fragments, the recovery of the S-S bonds by mercaptoethanol, the enzymatic hydrolysis by glutamyl endopeptidase from Bacillus intermedius and HPLC division of the resulting peptides were carried out. Attribution of the peptide fragments formed due to hydrolysis at the Glu-X bond in the ß-chain was accomplished by mass spectrometry. Mass spectrometry analysis data of the deuterium-labeled insulin samples' isotopomeric composition showed that the studied solid state isotope exchange reaction equally involved all the protein molecules. Biological studying of tritium-labeled insulin showed its physiological activity to be completely retained.

[Precursors and propeptides of neurotrophic factors as the modulators of biological activity of its mature forms].

Here, we review the problems of neurotrophic factors' folding, the role of its precursors (proneurotrophins) and the contribution of elements deleted during its maturation (propeptides) in biological functioning of these growth factors.

[Effect of a modification of the processing site of Bacillus intermedius glutamyl endopeptidase on the production of active enzyme by Bacillus subtilis cells].

A site-directed modification in position P1 of the processing site of Bacillus intermedius glutamyl endopeptidase was carried out. Variants of the protease gene were obtained that correspond to the protein with an Ala, Asn, Ser, or Glu residue in this position substituted for the Lys residue. The residue in the P1 position of the processing site was shown to affect substantially the production of the active enzyme; however, none of the mutations leads to the complete termination of active protein production by cells.

Hetero- and auto-activation of recombinant glutamyl endopeptidase from Bacillus intermedius.

Glutamyl endopeptidase from Bacillus intermedius (BIGEP) is a secretory serine proteinase specifically hydrolyzing peptide bonds involving alpha-carboxyl groups of glutamic and aspartic acids. In this work, different BIGEP forms (full-length precursor, precursor without signal peptide and mature part) were expressed in Escherichia coli and the process of enzyme maturation was studied in vitro. BIGEP precursor renaturation leads to autocatalytic hydrolysis of the propeptide at Glu(-16). At the same time, the enzyme activation requires the complete removal of the prosequence by other proteinases. The mature part of BIGEP cannot be activated, which indicates that the propeptide is required for the active protein formation. The data obtained allowed us to apply directed mutagenesis of the processing site to obtain a BIGEP form that matured autocatalytically. This approach makes it possible to produce the enzyme without extrinsic proteinases, which is a prerequisite for using it in limited hydrolysis of proteins and peptides.

[Oligomeric organization of recombinant human neurotrophins expressed in Escherichia coli cells].

Genes of human neurotrophins NGF, BDNF, NT-3 were cloned, and the corresponding proteins and their fragments were expressed in Escherichia coli BL-21 (DE3lambda) cells. Their intracellular localization was determined. The conditions for isolation and purification of the target recombinant proteins and for folding of BDNF and NT-3 precursors were selected. The recombinant proprecursors of human neurotrophines have been shown to possess complex oligomeric structure.

[Effect of deletion of 3'-noncoding region of the Bacillus intermedius glutamyl endopeptidase gene on the active protein production level in the culture of the B. subtilis cells].

The Bacillus intermedius glutamyl endopeptidase is a secretory serine proteinase from the subfamily of chymotrypsin. Its gene was previously cloned and sequenced. The enzyme was thoroughly characterized including 3D structure determination. The present work demonstrates that removal of 3'-noncoding region of the enzyme gene resulted in a decrease of the active glutamyl endopeptidase production level in culture of B. subtilis cells. In this 3'-noncoding region, the sequence with all typical features of transcription terminators of the Firmicutes type was found.