Biochemistry (Moscow), Supplement Series B: Biomedical Chemistry最新文献

Therapeutic peptides are an interesting class of pharmaceutical compounds that are structurally intermediate between small molecules and proteins but biochemically and therapeutically distinct from both. Currently, peptide-based drugs are used to treat immunological diseases, infections, and endocrinological and oncological diseases. The expression of therapeutic peptides in bacterial systems, for example, in E. coli, has a large number of advantages, such as high specific productivity, low production costs, great optimization potential, and good knowledge of the process. However, the creation of bacterial producer strains expressing therapeutic peptides with a molecular weight of less than 5 kDa is an extremely complex and time-consuming task due to the rapid degradation of the product by cell proteolytic enzymes or the high cytotoxicity of the target product. To solve such problems, the peptide is expressed in the form of tandem repeats or as a hybrid protein fused with another large protein, for example, SUMO. In this work, four peptide antigens differing in physicochemical properties were purified. The peptides were synthesized in soluble form as precursor proteins consisting of an N-terminal polyhistidine tag (His 6 tag), a SUMO fragment, and the amino acid sequence of the peptide. Hybrids were isolated from bacterial cells by disintegrating biomass under high pressure. The proteins were then purified by metal affinity chromatography using Ni-NTA agarose and enzymatically hydrolyzed with a highly specific SUMO protease. The resulting peptides were purified using metal affinity chromatography and final high-performance reverse phase chromatography on an SP-100-C8-PK sorbent. Next, the peptides were freeze-dried. As a result of work using the general technology, four peptides with different physicochemical properties were obtained. The purity by RP-HPLC was greater than 95%, and the mass of all peptides and fusion proteins was confirmed by HPLC-MS. The authenticity of the peptides was additionally confirmed by the ELISA method.

Malignant tumors represent a serious problem for humanity due to their widespread distribution and severe posttherapeutic consequences. The formation of metastases is the main criterion for tumor malignancy. A search for new compounds with antimetastatic activity remains an urgent problem. In this work, the antitumor activity of a cytolytic toxin Hct-S3 оf the sea anemone Heteractis magnifica was studied in an in vivo model of solid Ehrlich adenocarcinoma. It was established that intraperitoneal administration of Hct-S3 at doses of 0.01 and 0.02 mg/kg has hardly any effect on the tumor growth rate but prevents its metastasis from the primary tumor localization to other tissues. A significant decrease in the proliferative capacity of tumor cells was observed in the groups of mice treated with 0.01 and 0.02 mg/kg Hct-S3: the area of the tumor fluorescence zone decreased by 40 and 48%, and the index of integrated fluorescence density increased by 134 and 146%, respectively, as compared with animals without therapy. The obtained results confirm a high antiproliferative and antimetastatic potential of Hct-S3, which allows it to be considered as a promising compound for creating antitumor drugs.

The review presents an analysis of literary data on the role of microRNAs in the pathogenesis of type 2 diabetes mellitus and use in diagnostics as potential biomarkers and existing modern molecular genetic methods for the detection of microRNAs. The main functions of microRNAs in glucose metabolism are regulation of key enzymes of glucose catabolism, inhibition of activators/repressors of insulin transcription and genes that should not be expressed in β-cells, reduction of fusion and release of insulin granules, and influence on enzymatic reactions in peripheral tissues. The methods for determining microRNA expression levels were analyzed: Northern blotting, microarrays, NGS sequencing, RT-PCR, and ligation-mediated PCR. The results of the review will be useful for the development of microRNA-based diagnostic strategies for type 2 diabetes mellitus.

As a result of application of hydrogels with 0.05 and 0.005% HCRG21 calcipotriol-induced inflammatory swelling and flaking of the auricle were significantly reduced. As a result of application of hydrogels with 0.05 and 0.005% HCRG21 calcipotriol-induced inflammatory swelling and flaking of the auricle were significantly reduced. Treatment with HCRG21 normalized peripheral blood parameters in mice, including total white blood cell count and basophil percentage. In addition, treatment with gels containing HCRG21 resulted in a decrease in the level of proinflammatory cytokines: interleukin-23 (IL-23) and macrophage-derived chemokine (MDC) by 1.5 and 2 times, respectively, compared to the control group of animals. According to the results of histological analysis of the auricle of mice, treatment with peptide-containing gels enhanced the regeneration of the affected skin area, reducing hyperkeratosis, hyperplasia of the epidermis and dermis, and tissue hypervascularization. Thus, the discovered anti-inflammatory and regenerative effect of HCRG21 in the calcipotriol-induced allergic dermatitis model determines the potential of the peptide as a dermatotropic drug.

The unregulated and inappropriate use of antibiotics triggered the rapid spread of antimicrobial resistance in pathogenic microorganisms such as Pseudomonas aeruginosa. Antimicrobial peptides (AMPs) require investigations of their remarkable bioactivities and unique microbicidal mechanisms as potential replacement treatments for conventional antibiotics. Specifically, a naturally occurring antimicrobial dodecapeptide named Histatin 8 (HS8), from the Histatin family, has weak to moderate antimicrobial activities but is a promising bioactive molecule. In this study, three well-represented HS8-derived peptides with potential in silico bioactive propensities namely HS8-1 (Tyr12Phe), HS8-2 (Ser8Phe and Tyr12Phe), and HS8-3 (Gly11Phe) were explored. Results showed that HS8-1 was a significantly more active AMP than HS8-2 and HS8-3, displaying 3.0-fold and 3.56-fold decrease in minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values, respectively, against wild-type P. aeruginosa. Furthermore, HS8-1 and HS8-2 displayed the lowest MIC and MBC values against the antibiotic-resistant P. aeruginosa. Both HS8-2 and HS8-1 are the more active peptides showing inhibitory activities against wild-type and antibiotic-resistant P. aeruginosa biofilms in vitro, respectively. HS8-1 and HS8-2, at their MICs, can both primarily induce surface roughening and corrugation suggesting possible disruptive effects on the P. aeruginosa membrane reflecting antimicrobial action. HS8-1 lyses human red blood cells (RBCs) in vitro at concentrations 250 µg/mL and below, suggesting it to be the least hemolytic peptide compared to HS8-2 and HS8-3 which strongly lyse RBCs at relatively lower concentrations. Overall, the results imply that the HS8-derived peptide bioactivities can be enhanced by site-specific phenylalanine (Phe) substitutions against P. aeruginosa.

Atranorin is one of the main secondary metabolites of lichens of the group of depsides, which are of considerable interest due to their biological properties. Recently, its antioxidant and antiglycating properties, which may be important in the fight against the aging process and the development of age-related diseases, such as diabetes and neurodegenerative disorders, have attracted the active attention of researchers. The antioxidant action of atranorin is due to the neutralization of free radicals, which prevents oxidative damage to cells. Atranorin also exhibits antiglycation properties suppressing protein glycation, which can reduce the risk of complications in metabolic disorders.

A comprehensive study was conducted on the effect of vitamins D and K on some indicators of lipid peroxidation and the overall antioxidant activity of blood plasma in rats of two age groups. Differences in the studied parameters were revealed both in terms of age and under the influence of administered vitamins. An antioxidant effect of vitamin D is noted in adult animals, which is expressed in a significant decrease in the level of malondialdehyde (MDA) in blood plasma. In young people, a similar effect was found with the combined action of vitamins D and K. The content of lipids, glycogen, and cationic proteins in blood neutrophils of animals receiving vitamins D and K separately and together was analyzed. Significant changes in the content of cationic proteins were revealed when vitamin K was administered to adult animals. When analyzing the leukocyte formula, vitamin K and a complex of vitamins D and K showed the greatest effect, reflected in a change in the proportion of monocytes and lymphocytes. A negative correlation was shown between the lipid content in blood neutrophils and the level of one of the lipid peroxidation products, MDA, in blood plasma.

In the modern world, scientists often discuss problems associated with complications of pregnancy and childbirth in women. The study of cases of repeated fetal loss, stillbirth, intrauterine growth retardation and preeclampsia leads to the discovery of new aspects of this pathology. Of particular interest is Upshaw−Schulman syndrome, a rare congenital form of thrombotic thrombocytopenic purpura caused by mutations in the ADAMTS13 gene. This gene encodes a metalloproteinase capable of cleaving von Willebrand factor, which is important for blood clotting processes. A review of modern Russian and foreign, primarily English-language, literature was conducted on methods of diagnosis and treatment of congenital thrombocytopenic purpura as well as the prevention of complications of pregnancy and childbirth. This review highlights the main mechanisms of development and progression of the syndrome, current directions of management of women with Upshaw−Schulman syndrome, and methods of therapy of associated reproductive failure, and also presents modern groups of pharmacological drugs of choice. Pregnancy with Upshaw−Schulman syndrome is becoming a major focus of research in obstetrics, and it is important to conduct further high-quality association studies to develop innovative therapeutic options and options in the future.

Multipotent mesenchymal stromal cells (MSCs) are key regulators of tissue healing after injury. Acting in a paracrine manner, MSCs promote tissue regeneration and prevent the development of fibrosis, a pathological condition in which there is excessive replacement of functional tissue with scar tissue. However, the mechanisms of the antifibrotic action of proteins secreted by MSCs have not yet been established. The authors previously showed that the fraction of soluble protein factors (SF) of the MSC secretome is capable of suppressing the key cellular mechanism of fibrosis development, the differentiation of fibroblasts into myofibroblasts, and also established, using proteomic analysis, that this fraction is enriched in proteins capable of regulating the canonical signaling pathways of transforming growth factor beta, Wnt, and Notch in fibroblasts. This study investigated the effect of the SF fraction on these signaling pathways in human skin fibroblasts and found that this fraction suppresses the canonical Wnt signaling pathway in a model of myofibroblast differentiation of fibroblasts. According to the results of proteomic analysis, SF is enriched in the DKK3 protein, which is able to regulate fibroblast differentiation through interaction with components of the canonical Wnt signaling pathway. It was shown that depletion of DKK3 protein in the SF fraction of MSC secretome by immunoprecipitation results in a significant reduction in the ability of the SF fraction to suppress fibroblast differentiation into myofibroblasts in vitro. The obtained data clarify the mechanism of the antifibrotic action of MSC secretome proteins associated with the suppression of differentiation of fibroblasts into myofibroblasts.