Doklady Biochemistry and Biophysics最新文献

Intravital bioimaging based on luminescence is an important method for the development and testing of antitumor drugs on model animals and is an essential part of preclinical studies. Bioimaging based on luminescent systems, compared with fluorescent bioimaging, provides a high signal-to-noise ratio, which justifies the development of cell lines that stably express luciferase genes for their subsequent use in model animals. In this work, we describe the creation of a stable cell line SKOV3.ip1-NanoLuc constitutively expressing the NanoLuc luciferase gene. The developed cell line was shown to be effective for intravital luminescence bioimaging of immunodeficient animals with deep-seated intraperitoneal tumors, which can be considered as a model of late-stage ovarian cancer.

Cellular proteins, partners of viral enzymes, are involved in the replication of human immunodeficiency virus type I (HIV-1) at various stages. Thus, the viral enzyme integrase, participating in several stages of the viral cycle, interacts with various cellular proteins. A number of them are already known, and for some the mechanism of action has been established, but the search for cellular partners of integrase continues. In this work, the identification of cellular partners of HIV-1 integrase has been carried out by the method of cross-linking followed by mass spectrometry (XL-MS). Twelve new potential integrase partners have been identified, and some of them have been examined for their effect on the early stages of HIV-1 replication.

The aim of the study was to investigate the correlation between cytokine levels and values of antibodies to cyclic citrullinated peptide (ACCP) and antibodies to carbamylated proteins (anti-CarP) in patients with rheumatoid arthritis (RA).

. A total of 106 patients with a reliable diagnosis of rheumatoid arthritis were included in the study. Determination of anti-CarP and anti-CCP was performed by enzyme immunoassay. Patients were divided into subgroups depending on the values of ACCP and anti-CarP. The concentration of 27 cytokines in serum was determined using multiplex xMAR technology.

. When comparing immunological subgroups, ACCP(+) patients had higher concentrations of IL-1β, IL-1Ra, IL-2, IL-6, IL-8, IL-10, IL-12, IL-13, IL-15, IL-17, FGF, G-CSF, GM-CSF, IFN-γ, IP-10, MCP-1, MIP-1α, TGFb, TNF-α, and VEGF. IL-5, IL-9, eotaxin, MIP-1β, and RANTES values were higher in ACCP(–) patients. In the subgroup of ACCP(–) patients, an inverse correlation between IL-5 and total Sharpe score and between IL-9 and DAS28-CRP was found. In anti-CarP(–) patients (n = 73), higher values of IL-17 were recorded.

. Our data support the concept of RA heterogeneity, characterized by the existence of different clinical and immunological subtypes, which may have implications for improving personalised therapy.

The composition and content of fatty acids of the sand lizard (Lacerta agilis) and its potential food objects in the shore and steppe biotopes were studied. For the first time, it was established that the content of long-chain omega-3 polyunsaturated fatty acids, primarily eicosapentaenoic acid (EPA), significantly increases in the tissues of lizards from a shore biotope during the period of emergence of amphibious insects from the nearby saline Lake Shira. Among all the food sources of lizards, the highest EPA content was found in chironomid imago. The content of docosahexaenoic acid (DHA) in the muscles of the lizards was also high, although this acid was not detected in the invertebrates that the lizards consumed. In this regard, it was assumed that the sand lizard is able to biosynthesize DHA from biochemical precursors contained in food.

Enzymatic inhibitory assay based on the coupled enzyme system NAD(P)·H:FMN oxidoreductase and luciferase (Red + Luc), originally developed for environmental monitoring of soils, water, and air, is proposed as a method for evaluating changes in the properties of active ingredients of pesticide preparations depending on the additional components (formulants), as well as when pesticides are combined in mixtures. Using the commercial pesticide preparations containing glyphosate, it was shown that the degree of inhibition of the coupled enzyme system Red + Luc largely depends on the formulants rather than on the active ingredient in their composition. Moreover, the combined inhibitory effect of the pesticides mixture on the coupled enzyme system Red + Luc was not additive. According to the results of the inhibitory assay, the type of interaction of pesticide preparations in mixtures depends on both the formulants used and the ratio of pesticides in the mixture.

The objective of this work was to study the number of different T-helper subpopulations in peripheral blood as well as the expression of TBX21, RORC, GATA3, NFKB1, MAPK8, and STAT3 genes, responsible for the regulation of T-helper differentiation in persons chronically exposed in utero. The object of the study was peripheral blood cells obtained from 156 people chronically exposed in utero and postnatally in a wide range of doses on the Techa River. The mean cumulative absorbed dose to the red bone marrow in the examined exposed individuals was 496 ± 51.2 mGy (dose range, 73.5–1298 mGy), in the comparison group 1 it was 18.7 ± 1.97 mGy (dose range, 0.78–57 mGy), and in comparison group 2 (exposed only postnatally) it was 571 ± 49.1 mGy (dose range, 86.74–1240 mGy). The subpopulation composition of T-helper cells was analyzed by flow cytometry. The relative mRNA content of TBX21, RORC, GATA3, NFKB1, MAPK8, and STAT3 genes was assessed by RT–PCR. A dose-dependent decrease in the total number of T-helper cells, effector memory T-helper cells, and central memory T-helper cells at a trend level and an increase in the relative number of naive T-helper cells in the peripheral blood of individuals exposed in utero and postnatally were found. An increase in the relative number of type 1 T-helper cells was also revealed in those exposed in utero and postnatally relative to the group of persons exposed only postnatally. These changes did not depend on the cumulative radiation doses. There were no statistically significant changes in the mRNA expression of the studied genes (GATA3, STAT3, TBX21, MAPK8, and RORc). No correlation between the mRNA expression of the studied genes and the relative number of cells in the subpopulations of T-helper types 1, 2, and 17 in the examined individuals was revealed.

The objective is to conduct a pilot study on the quantity of regulatory T-cells (Treg) in the peripheral blood and to assess transcriptional activity of FOXP3 gene in chronically exposed persons. Materials and methods. The study included 77 participants who were divided into two groups: exposed people—45 individuals, with a mean cumulative dose to red bone marrow (RBM) of 641.21 ± 80.41 mGy, and a comparison group—32 individuals, with a mean cumulative RBM dose of 20.38 ± 2.51 mGy. The study on the assessment of the FOXP3 gene expression was conducted in 298 individuals: the exposed group consisted of 163 individuals with a mean cumulative dose to RBM of 702 ± 43.10 mGy; the comparison group included 135 individuals with a mean cumulative dose to RBM of 17.30 ± 1.40 mGy. The study groups did not differ significantly in age, sex, and ethnicity. Quantitative assessment of regulatory T-cells in the peripheral blood was performed using flow cytometry method by the presence of T-helper markers CD3 and CD4, high expression of marker CD25 and low expression of marker CD127. Thus, the phenotype of regulatory T-lymphocytes was described as CD3+CD4+CD25highCD127low. The relative mRNA content of the FOXP3 gene was assessed by PCR-RT. Results. More than 70 years after the onset of chronic exposure, no statistically significant changes in the pool of regulatory T-cells were detected in the exposed persons: the content of absolute and relative number of Treg did not differ statistically significantly between the studied groups (p = 0.91 and p = 0.29, respectively); no statistically significant relationship between Treg indices and the cumulative doses to RBM and thymus and peripheral lymphoid organs were found. No statistically significant differences in FOXP3 gene mRNA expression were found between exposed individuals and the comparison group. A linear positive dependence of FOXP3 gene mRNA expression on the relative number of regulatory T-cells was shown (p = 0.007).

Objective. To study the repertoire of the T-cell receptor in persons chronically exposed to radiation in the long-term period. Materials and methods. The study involved 48 people, who were divided into two groups: a group of exposed persons (31 individuals with the average accumulated dose to red bone marrow (RBM) of 981 ± 130 mGy) and a comparison group (17 individuals with the average accumulated dose to RBM of 25.3 ± 5.91 mGy). The study groups did not differ significantly in age, gender and ethnicity. The repertoire of Vβ-segments of the T-cell receptor of the peripheral blood T-lymphocytes of exposed persons was analyzed by flow cytometry method. 24 Vβ-segments of the T-cell receptor were studied. Statistical processing of the obtained data was carried out using the Wilcoxon signed-rank test, and a direct description of Vβ-segment repertoire of the T-cell receptor was performed using the Lorenz curve and the Gini-TCR index. Results. The study revealed a statistically significant increase in the number of Vβ3 and Vβ5.2 T-cell receptor segments in exposed individuals relative to the comparison group (p = 0.03 and p = 0.003, respectively). It was also shown that the distribution of the Vβ-segments of the T-cell receptor is uneven in both study groups. However, there was no significant difference between the repertoires of the T-cell receptor of the studied groups in the Gini-TCR index (p = 0.14).

The efficiency of DNA integrity repair processes after radiation exposure may depend on hereditary variations of repair genes caused by single nucleotide polymorphisms. Disturbances or even failure of repair processes trigger a chain of reactions leading to genome instability and oncogenic transformation of the cell. Purpose. To investigate the association of single nucleotide polymorphism in genes of nucleotide excision repair (ERCC2 rs13181, XPC rs2228001), AP site repair (APEX rs1130409), homologous recombination (XRCC3 rs861539), single-strand DNA break repair (XRCC1 rs25487), and double-strand DNA break repair (PARP rs1136410, XRCC4 rs2075685) with the risk of development of malignant neoplasms of various localizations in chronically exposed persons. Materials and methods. The study was perfomred in 861 individuals who were chronically exposed to low-dose low-rate radiation, 274 of which had malignant neoplasms (MN) of various localizations and 587 made up the comparison group (exposed persons without MN). The mean cumulative dose to red bone marrow (RBM) in the group of people with MN was 561.65 ± 25.31 mGy, while in the comparison group it was 543.14 ± 36.06 mGy. Genotyping of polymorphic loci rs13181, rs2228001, rs1130409, rs861539, rs25487, rs1136410, and rs2075685 was performed by real-time PCR. The association of polymorphic loci with the risk of MN development was determined by the odds ratio (OR) and 95% confidence interval (95% CI). A multifactor dimensionality reduction method was used to assess intergenic interactions. Results. Single-stranded DNA break repair gene (XRCC1) rs25487 polymorphism in accordance with the dominant model is associated with an increased risk of MN development in the combined group of the examined persons (OR = 1.79 (1.12‒2.87), p = 0.01) and in the Slavs group (OR = 2.26; 95% CI 1.06-4.81; p = 0.03). The rs861539 polymorphism of the gene involved in homologous recombination (XRCC3) in accordance with the recessive model is associated with a reduced risk of MN development both in the combined group of exposed persons (OR = 0.25 (0.15‒0.41); p < 0.00001) and separately in the group of the Slavs (OR = 0.28 (0.13‒0.60); p < 0.0001) and in the group of the Turkic people (OR = 0.22 (0.11‒0.44); p < 0.0001). The model of interfactorial interactions allowed us to establish a protective effect with respect to the risk of MN development in the carriers of polymorphic loci rs861539 of the XRCC3 gene and rs1130409 of the APEX1 gene (p < 0.001).