Sex differences were studied under normal conditions and after a single mild stress in inbred Wistar rats. It was found that intact females had higher blood corticosterone and leptin levels than males. In females, locomotor activity and the number of active defense reactions in elevated plus maze were higher, while the number of passive defense reactions was lower than in males. After stress, the blood corticosterone and leptin levels increased in both females and males, while the level of the main sex hormones decreased. We found for the first time that leptin and corticosterone levels positively correlated with anxiety index, locomotor activity, and number of active defense reactions and negatively correlated with testosterone and estradiol levels. A hypothesis was put forward that leptin, together with corticosterone, is involved in the organization of the adaptive behavior under stress conditions.

The autophagy-related structures, the size of the rough endoplasmic reticulum (ER) cisterns, and the expression of apoptosis-related proteins were assessed by transmission electron microscopy and immunohistochemistry in tumor samples from mice with B16 skin melanoma after administration of autophagy inducer (rapamycin) or ER stress inducer (brefeldin A). Brefeldin A stimulated significant ER stress in mouse skin melanoma cells, but rapamycin contributed to the maintenance of cell homeostasis by inducing autophagy, which was confirmed by the presence of autophagy-related structures and significantly smaller sizes of the rough ER cisterns in the group of mice receiving both rapamycin and brefeldin A. Brefeldin A-induced ER stress triggered apoptosis of tumor cells. Moreover, simultaneous stimulation of autophagy and ER stress in tumor cells promoted cytoprotective selective autophagy (reticulophagy) aimed at resolving ER stress, which may be a mechanism underlying the development of chemoresistance in skin melanoma.

The biological properties of nine clinical isolates of Pseudomonas aeruginosa obtained from five patients with cystic fibrosis were assessed. Six different sequence types of P. aeruginosa with different biofilm-forming abilities were detected. It was found that at the initial stage of adhesion, four isolates were tobramycin-resistant, while in the formed biofilm, almost 100% survival was recorded. The properties of the isolates were associated with the sequence type: when it changed, the biofilm-forming ability decreased, but the number of viable cells resistant to tobramycin at a concentration of 2 μg/ml increased. The study shows that the existence of a biofilm is a key factor ensuring, despite constant antimicrobial therapy, the long-term persistence of P. aeruginosa in patients with cystic fibrosis.

We performed a comparative analysis of lipid and cholesterol metabolism disorders, as well as liver condition in male Wistar rats receiving high-fat diets with different cholesterol contents. Addition of 2% cholesterol to the diet resulted in a marked aggravation of fatty liver disease, characterized by extreme accumulation of fat and cholesterol. A diet supplemented with 0.5% cholesterol resulted in serum cholesterol accumulation and liver steatosis, but had no effect on weight gain or body composition. Introduction of 1% cholesterol to the diet induced similar abnormalities as 2% cholesterol, but without the extreme cholesterol and fat accumulation in the liver. This model can be used in the future to evaluate the effectiveness of the studied functional foods.

Among the species belonging to the genus Lysobacter, known as micropredators, there are valuable strains-producers of antibiotics that can be effective in the fight against pathogenic microorganisms. The morphological features (pigments and cell sizes) of a new strain Lysobacter sp. Hz25 were studied during cultivation on different media. It was found that the size of Lysobacter sp. Hz25 cells depends on the number of CFU per unit area of solid medium and the location relative to the center of the colony. At low CFU number and at the periphery of the colony, the cells are longer, which indicates their possible transition to a mobile state. The color of the colonies depended on the medium composition. The presence of a complex of yellow hydrophobic pigments in the cells and a complex of hydrophilic pigments secreted into the medium and coloring it pink and yellow was found. Understanding of the effect of the medium composition on the morphology of Lysobacter sp. Hz25 cells and colonies and, consequently, on its interaction with pathogenic microorganisms will allow more complete use of its properties for combating human infections.

The protective properties of argon-oxygen mixture were studied in the culture of M-22 human fibroblasts, piglet thymus cells, and rat brain cells. To induce damage to cell membranes, 200 μM HO and 500 μM acetic acid were used. The cells were exposed in the presence of a gas mixture containing 70% argon and 30% oxygen (ArgOx 70/30) at 37°C for 6 h. The ArgOx 70/30 mixture produced no toxic effect on the cells of all studied cultures. In culture of M-22 fibroblasts, acetic acid-induced cell death decreased by 1.5-1.9 times under the influence of ArgOx 70/30 and the cell membranes remained better preserved. In none cultures, ArgOx 70/30 mixture significantly protected cells from hydrogen peroxide-induced damage. Piglet thymus culture cells demonstrated very high resistance to high concentrations of hydrogen peroxide and acetic acid, while rat brain cells, on the contrast, had low resistance.

Tuberculous meningitis (TBM) is the most prevalent and severe manifestation of tuberculosis in CNS. The mechanisms of neurological injury caused by TBM are not well understood. Our study showed that overexpression of WTAP (Wilms tumor 1 associated protein) reduced inflammatory factors and Iba-1 expression induced in BV-2 by H37Rv. It also increased proliferation of neural stem/progenitor cells (NSPCs) and expression of the neuronal marker DCX in these cells. WTAP enhanced expression of high mobility group nucleosome-binding domain-containing protein 3 (HMGN3) by promoting mA methylation of its mRNA. Reducing HMGN3 expression negated WTAP-induced anti-inflammatory and neuroprotective effects in TBM cell model. WTAP inhibited inflammation and microglia activation while promoting NSPC differentiation into neurons via elevation of HMGN3 expression. WTAP/HMGN3 proteins and the corresponding mRNA could be potential targets in the treatment of TBM.

This review considers cell therapy and possible mechanisms underlying beneficial effects of cells in prevention of neurodegenerative changes as a promising approach to the treatment of Alzheimer's disease. The results of using various types of cells in experimental models of Alzheimer's disease and their effect on the regeneration of the brain and recovery of cognitive functions are presented.

Markers of α2- and β1-adrenergic receptors (AR) were detected immunohistochemically in the liver of db/db mice with obesity and type 2 diabetes mellitus before and after melatonin treatment. Melatonin (1 mg/kg in 200 μl distilled water) was administered intragastrically from the 8th to the 16th week of life. The comparison groups were intact and placebo-treated db/db mice. Melatonin administration resulted in a significant increase in the relative areas of β1- and α2-AR expression, with a tendency towards an increase in the area ratio, as well as a significant increase in the ratio of β1/α2-AR concentrations due to preferential increase in β1-AR parameters. Melatonin administration apparently reduces sympathetic neuropathy of the liver and promotes the shift of lipid metabolism processes in hepatocytes towards lipolysis activation, which allows us to consider this hormone as a promising component of complex therapy of fatty liver disease.

It was shown that blocking of glycolysis by 2-deoxy-D-glucose (DG) inhibits LPS-induced NO production in glial cell cultures and reduces the mitochondrial membrane potential. Lactate partially prevented the decrease in the mitochondrial membrane potential of gliocytes, but did not affect the inhibitory effect of DG. Immunocytochemical typing showed the presence of a large number of astrocytes and microglia in the culture. In the control, microglial cells had a large number of processes, which is typical of nonactivated cells, whereas in LPS-treated cultures, microglia had a flattened amoeboid morphology typical of activated microglia. In cultures treated with LPS against the background of DG, both cells with processes and amoeboid cells were present. Our results indicate that inhibition of glycolysis is a strong modulating factor in inflammation.

Coiled-coil domain-containing protein 86 (CCDC86) expression is correlated with the occurrence of lymphoma. However, the expression of CCDC86 in solid tumors such as nasopharyngeal carcinoma (NPC) and the effects of CCDC86 on tumorigenesis remains unclear. Here we studied both problems using tumor tissue samples from NPC patients, NPC cell lines (in vivo), and a model of transplanted tumor in BALB/c nude mice (in vitro). We found that CCDC86 protein was expressed in all studied cell lines, but its expression in CNE1, CNE2, CNE-2Z, 5-8F, and 6-10B cell lines was higher than in nasopharyngeal epithelium cell lines NP69 and NP460. In tumor tissues obtained from patients with NPC, CCDC86 expression was higher than in normal (adjacent) tissues. Knockdown of CCDC86 gene inhibited colony formation and cell proliferation, but increased apoptosis. In BALB/c nude mice transplanted with CCDC86-knockdown CNE-2Z cells, tumors barely grew in comparison with the controls transplanted with CNE-2Z cells transfected with an empty vector lentivirus. In conclusion, CCDC86 is expressed in NPC tissues and NPC cell lines and is closely associated with NPC tumorigenesis. Our study may provide insights into exploring the novel therapeutic targets for NPC.

It is known that the senescence-associated secretory phenotype (SASP) can promote senescence of surrounding normal cells. However, SASP signaling during senescence of mesenchymal stromal cells (MSCs) has not yet been fully studied. We determined the pattern of secreted proteins specific to MSCs under stress-induced senescence. Using chromatography-mass spectrometry, proteins specific to the secretome of senescent or "young" cells were identified. The secretome of senescent cells contains proteins both associated with senescence (LOXL2, CCL2, PLAT, SERPINE2, etc.) and important for reducing the impact of these processes, in particular, proteins responsible for inhibition of oxidative stress (MIF, PRDX5, GSTM2), detoxification of methylglyoxal (GLO1), and suppression of inflammatory reactions (GAS6, GSTM2). The obtained results indicate the complex etiology of aging and the ambiguity of the function of SASP within the paracrine induction of aging of neighboring cells.

The mitotic index is a critical indicator of the proliferative activity of cell populations and is widely used in oncology and stem cell research. One of the most promising methods for its assessment is imaging flow cytometry implemented on the Amnis ImageStream platform using the IDEAS software. A critical evaluation of the built-in Wizards, Cell Cycle-Mitosis algorithm revealed several limitations, including a high degree of operator-dependent variability in gate setting and difficulties in identifying the mitotic population in the absence of distinct peaks on the cell cycle histogram. An alternative approach, the Mean + xSD algorithm, was proposed. This method is based on automated quantitative assessment of the Bright Detail Intensity R3 parameter and excludes the need for manual gating and histogram interpretation. Using the Caco2 and HT-29 cell lines, we demonstrated that the proposed algorithm exhibits accuracy comparable to the classical IDEAS algorithm, and in some cases provides even more reproducible quantification of the mitotic index. These results demonstrate the potential of the new algorithm as a more objective and robust tool for analyzing mitotic activity in cultured cells.

The sizes of intracerebral high-grade gliomas 101.8 and C6 in Wistar and Long-Evans rats and their relationship with survival were determined; changes in the thickness of the cerebral cortex were evaluated and morphological features of glioma models were studied. Median glioma volumes measured on MRI of brain slices in Wistar rats with glioblastoma 101.8 and glioma C6 were 169 (148-252) mm and 114 (47-154) mm, respectively; in Long-Evans rats with glioma C6, the median volume of the tumor was 159 (85-223) mm without significant differences. The mean survival time of Wistar rats with glioblastoma 101.8 and glioma C6 were 16 ± 1 (SE = 0.3) days and 33 ± 6 (SE = 2) days; in Long-Evans rats with glioma C6, the mean survival time was 30 ± 2 (SE = 4) days. Survival in rats with glioblastoma 101.8 was significantly lower than in animals with C6 glioma. A negative correlation of tumor size and survival was revealed in Wistar and Long-Evans rats with glioma C6: r = -0.80 (p = 0.006) and r = -0.70 (p = 0.03), respectively. The mean thickness of the cortex of the contralateral hemisphere in tumor-bearing rats was significantly lower than in sham-operated animals. Glioma models vary in the growth rate. At the late stages of 101.8 and C6 gliomas, the thickness of the cerebral cortex of the contralateral hemisphere decreases. The volume of C6 glioma can be used as a predictor of Wistar and Long-Evans rat survival.

We evaluated the response of Mycobacterium tuberculosis to exposure to a new aroylhydrazone derivative using the in vitro mutagenesis followed by genomic analysis of a resistant variant. The compound N'-[(E)-(5-methoxy-1H-indol-3-yl)methylidene]furan-2-carbohydrazide showed minimal inhibitory concentration (MIC) of 0.4412 μM for the reference strain M. tuberculosis H37Rv. An H37Rv clone resistant to elevated (4 × MIC) concentration of this compound recovered on a solid medium was further analyzed by whole genome sequencing and bioinformatics tools. A non-synonymous mutation was detected in the Rv3755c gene at position 302A>G (codon 101H>R, CAC-CGC). The gene-gene interaction analysis showed that this gene belongs to a network that also includes several ABC transporter genes. The identified mutation in Rv3755c may be associated with bacterial adaptation to the selective pressure of the studied aroylhydrazone derivative and reflect a non-specific drug tolerance mechanism. The conserved M. tuberculosis protein Rv3755c, whose function is unknown, may be related to the ABC transporter efflux system.

The study examined implication of miRNA-19b carried by endothelial microparticles (EMPs) in phenotypic switching of vascular smooth muscle cells (VSMCs) and the mechanisms underlying this transformation. The functions of miRNA-19b were assessed by phenotypic switching, proliferation, and migration of VSMCs. The target genes of miR-19b associated with proliferation and migration were revealed by analysis of TargetScan and miRanda databases and verified with luciferase assay. Experiments showed that EMPs could transfer miRNA-19b into VSMCs. Elevated content of miRNA-19b increased expression of contractile phenotypic markers SMA and SM22α and inhibited proliferation and migration of VSMCs. The direct target gene of miRNA-19b turned out to be the mitogen-activated protein kinase 6 (MAPK6). Thus, miRNA-19b in EMPs could inhibit phenotypic transformation of VSMCs from the contractile phenotype to synthetic one and reduce proliferation and migration by down-regulating MAPK6 expression, which can potentially inhibit the development of atherosclerosis.

The TNFα content in the blood serum of 109 patients with uterine fibroids was studied. Cytokine concentrations ranged from 4.65 to 25.88 pg/ml regardless of the clinical parameters of the disease, but were associated with complex genotypes of different structural and functional organization (TNFA, IL1B, IL4, IL6, IL8, IL10, IL17A, VEGFA, MMP2, MMP3, MMP9). The interaction of multiple genes that affect the level of serum TNFα production has been revealed. In particular, on the one hand, the complex genotype TNF-308GG/VEGF-2578CA is positively associated with high levels of TNFα (OR = 10.00, p = 0.0274), and on the other hand, the combined genotype of 4 polymorphisms TNF-308GG/TNF-238GG/VEGF-2578AA/MMP2-1306CC is closely associated with its low level (OR = 25.00, p = 0.0469). The obtained results allow better understand the mechanisms of the manifestation of the genetic predisposition to uterine fibroids associated with the presence in the genome of highly specific gene compositions that underlie tumorogenesis in this disease.

We studied the effects of female sex hormones estradiol and progesterone on the expression of CCR5 and CCR8 co-receptor genes (that play an important role in the HIV-1 entry into the cell) in human peripheral blood mononuclear cells (PBMC) isolated from different female donors and infected with HIV-1 subtype G. Female sex hormones produced a dose-dependent effect on the replication of HIV-1 subtype G: low doses of estradiol and high doses of progesterone significantly induced CCR8 expression in PBMC of all donors, which correlated with an increase in viral load by 1.5-1.7 times on average. The exception was one donor, in whom a high dose of estradiol also induced an increase in CCR8 expression. High concentrations of progesterone also enhanced the expression of the CCR5 co-receptor. The detected differences in the co-receptor expression in infected PBMC from different donors indicates that the host genetics may also play an important role in PBMC susceptibility to HIV infection.

Metagenomics of bacterial communities in tuberculosis caseous necrotic mass indicates the predominance of facultative anaerobes. Nine strains isolated from the tuberculosis necrosis were identified to species and whole-genome sequencing was performed: Staphylococcus hominis (3 strains), S. epidermidis (3 strains), Corynebacterium ureicelerivorans (2 strains), and C. kefirresidentii (1 strain). Prediction of metabolic pathways and virulence factors showed that Corynebacterium and Staphylococcus possess gene sets that are absent in Mycobacterium tuberculosis: lipases and proteases for the degradation of caseous necrosis, glutamate and polysaccharide capsules, ureases capable of increasing pH of the caseum; and Fe(III) uptake systems. The isolated species can form a bacterial consortium with M. tuberculosis at the early (Corynebacterium) and later (Staphylococcus) stages of necrotization of the tuberculosis focus in the lungs.

Escherichia phage Ec1-7 was characterized and its efficacy on food products, as well as on stainless-steel surfaces, was evaluated. The bacteriophage with high adsorption rate and short latency period remained stable when exposed to various aggressive factors and demonstrated lytic activity against STEC strains. In addition, the phage did not contain antibiotic resistance, virulence, and toxin genes. In 24 h after application of the bacteriophage, E. coli concentration on lettuce leaves and on stainless-steel surfaces decreased by 98.2 and 90.5%, respectively. In the experiment with artificially contaminated beef, an 81% decrease in bacterial concentration was recorded on day 3 in comparison with the control samples treated with 0.9% NaCl solution. The bacteriophage Ec1-7 exhibited stability and efficacy as a biocontrol agent across diverse test materials.

We studied the distribution and survival of mesenchymal stem cells (MSCs) transplanted within a fibrin hydrogel into the spinal cord of immunocompetent rats without spinal cord injury (SCI) and with contusion SCI within the first hours after injury. MSC migration was monitored by MRI, and cell survival was assessed by immunofluorescence and immunohistochemistry on spinal cord sections. It was shown that transplanted allogeneic MSCs remain viable for at least 7 days in case of intrafocal administration 30 min after experimental contusion SCI and for at least 28 days in case of subdural and intramedullary transplantation into the intact spinal cord. MSCs are predominantly located at the injection site. Thus, our data demonstrate that allogeneic MSCs transplanted into the SCI site in the acute phase can survive for at least 7 days without migrating into surrounding tissues.