Stress proteins dehydrins have been identified using specific antibodies in buds silver birch (Betula pendula Roth) growing in contrasting climatic conditions of the regions Karelia and Central Yakutia. Dehydrins in buds of plants are presented in two areas: medium molecular weight (56-73 kDa) and low molecular weight (14-21 kDa), and 17 kDa dehydrin was detected in all studied birches. 14-21 kDa dehydrins are the most susceptible to seasonal changes. Regardless of birch growing region, dehydrines 14-21 kDa show significant increase in their level in the autumn and winter when the trees are characterized by the highest frost resistances. Dehydrins have relealed intraspecific polymorphism, more pronounced in Yakutia plants in conditions of extreme continental climate, compared with Karelia, which is probably due to the peculiarities of woody plants to adapt to the extremely low winter temperature of the permafrost zone.

The aim of the present study was to compare the effect of sulfated fucopolysaccharides isolated from Fucus vesiculosus on cell lines HeLa G-63, Hep G2 and Chang liver. Native fucoidan F3 and two fractions (F3-0.5 and F3-1), obtained by anion-exchange chromatography, were analyzed using chemical methods and IR-spectroscopy. Their action on cells demonstrates that F3 and F3-1, characterized by higher content of sulfates, preferable location of sulfo-groups at C4 atom of fucose residue and low content of uronic acids, inhibit cell proliferation more efficiently. Human liver tumor cells Hep G2 appeared to be the most sensitive to fucoidan treatment whereas nonmalignant human cells Chang liver were the least sensitive.

One of the approaches to correct type 2 diabetes mellitus (T2DM) and its complications is the use of drug bromocryptine mesylate (BCM), a selective agonist of type 2 dopamine receptors (DA2R). At the same time, the efficiency and the mechanisms of action of BCM in treatment of severe forms of T2DM are not currently understood. The objective was to study the effect of four-week treatment of male rats with neonatal T2DM model using BCM (300 mg/kg/day) on their metabolic parameters and activity of the adenylyl cyclase signaling system (ACSS) in the hypothalamus. The BCM treatment restored glucose tolerance and its utilization by exogenous insulin, and normalized lipid metabolism by lowering the levels of triglycerides and atherogenic cholesterol increased in T2DM. In the hypothalamus of BCM-treated diabetic rats, the regulation of ACSS by agonists of type 4 melanocortin receptor (MC4R), DA2R and 1B-subtype serotonin receptor, and the expression of Mc4r gene encoding MC4R were restored. Meanwhile, the BCM treatment had no effect on plasma insulin level and insulin production by pancreatic b-cells. The obtained data indicate the significant prospects of BCM to treat severe forms of experimental T2DM, and show that the therapeutic potential of this drug includes its ability to restore the hypothalamic signaling systems sensitive to monoamines and peptide of the melanocortin family, which are responsible for the control of energy metabolism and insulin sensitivity.

Lipophilic derivatives of peptides corresponding to the cytoplasmic regions associated with the G-protein coupled receptors (GPCRs) are capable of functioning as an intracellular agonist. Previously, we have shown that peptides corresponding to region 562—572 of luteinizing hormone receptor (LHR) and modified by decanoate and palmitate at the C-terminus activate adenylyl cyclase (AC) in the testes of rats. The stimulating effect of peptide 562—572 modified by decanoates at the N- and C-termini (IV) peaked at a concentration of 10(–5) M and then subsequently decreased with increasing concentration. We hypothesized that this may be due to ability of the peptide IV to micelle formation. To test this suggestion, we examined the relationship between biological activity, hydrophobicity and ability to micelle formation for peptide IV and other acylated derivatives of peptide 562—572 including the derivatives containing C-terminal decanoate (III) and palmitate (VI). It has been shown that the stimulating effect of peptide IV at a concentration of 10(–5) M on AC activity in the plasma membranes of rat testes and ovaries is only slightly inferior to that of peptide VI and superior to the corresponding effect of peptide III. The effect of peptide IV at a concentration of 10–3 M was reduced by 20—27 % and amounted to 50—51 and 87—88 % of that of peptides VI and III, respectively. Despite the high hydrophobicity, the peptide IV had abnormally low retention time in reverse-phase HPLC when it was eluted from the Nucleosil C8 column, even lower than that of unmodified peptide 562—572. However, with increasing concentration of trifluoroacetic acid in the eluent from 0.1 to 0.5 % causing the destruction of micelle-like structures, the retention time of the peptide IV was significantly increased, whereas it remained unchanged in the case of the other peptides. Surface tension of aqueous solution of peptide IV insignificantly decreased with the increase of its concentration, but then, at peptide concentration of 710(–6) M, the sharp decline and the plateau were found, which indicates the beginning of the formation of micelles. Thus, at concentration of 10(–5) M and higher the peptide IV forms micelles which prevents its interaction with the receptor. The ability of GPCR-peptides to self-aggregation and micelle formation should be taken into account when developing their membrane-active analogues.

In this paper we investigate the distribution of myosin-9 in the cytoplasm of human embryonic lung fibroblasts. Using immunofluorescence, immunoprecipitation and western blotting, we demonstrated that myosin-9 forms multimolecular complexes with high molecular weight tropomyosin isoforms and actin in cytoplasm of cultured cells. We explored the levels of myosin-9, tropomyosin and actin in cytosol and cytomatrix extracts at different time points after LPA addition (20 ng/ml) to culture medium. Cytosole extracts from human embryonic lung fibroblasts were subjected to co-immunoprecipitation assay with polyclonal antibodies specific to C-terminal peptide of human myosin-9, or with monoclonal antibodies recognizing high molecular weight tropomyosin isoforms. The cross-immunoprecipitation method revealed changes in the composition of myosin- 9/tropomyosin complexes under the action of LPA. We have observed the significant decrease in myosin-9 co-immunoprecipitated tropomyosin level immediately (1 min) after LPA addition. Additionally, we have found that LPA treatment during 1 h induces proteolytic degradation of myosin-9 with accumulation of 130 kDa and 40 kDa fragments in cytomatrix fraction. These results suggest that cytoplasmic multimolecular protein complexes containing myosin-9 and tropomyosin are involved in the regulation of cellular response to LPA.

Nuclear lamins form nuclear lamina localized under the inner nuclear membrane. It was previously considered that the nuclear lamina predominantly plays a structural role, however, its involvement have been recently described in the regulatory processes such as chromatin organization and gene transcription. It is known that mutations in the LMNA gene lead to the development of a large number of diseases, laminopathies, which mainly affect mesenchymal tissue. Nowadays, the mechanisms by which the lamina can regulate cell differentiation remain incompletely understood. In the present work, we have studied the effect of LMNA gene mutations on the process of muscle differentiation of primary satellite cells and Ñ2Ñ12 cell line. The genome of satellite cells and Ñ2Ñ12 cell line was modified by the introduction of lentiviral constructs encoding LMNA G232E associated with the development of muscular dystrophy Emery—Dreyfus and LMNA R571S associated with the development of dilated cardiomyopathy. The morphology of the cells was estimated using immunofluorescence, the expression level of myogenic genes were analyzed by qPCR. We have shown that the analyzed mutations reduce the ability of cells to differentiate, to fuse and to form myotubes. We have suggested that it is due to enhanced expression of markers at the early stages and to reduced expression markers at the late stages of myogenesis. Therefore, mutations in nuclear lamins can influence the process of muscle differentiation.

Mechanisms of coenzyme Q10 effect on serum-deprived glioma cell proliferation have been studied. Our results have shown that the addition of coenzyme Q10 into serum-free culture medium leads to increase in cell viability, stimulation of cell growth, as well as restoration of mitochondrial potential and increase of quantity of energized mitochondria. It has been found out that coenzyme Q10-induced glioma cell proliferation under serum deficiency is a result of intracellular reduced glutathione concentration decrease with subsequent activation of proteinkinase C, ERK1/2 and phosphoinositol-3-kinase.

Cell cultures of higher organisms, especially cultures of human cells, are increasingly used in medical, pharmaceutical and scientific research. The main problem of cell cultures — non-lethal hidden contamination by mycoplasmas, viruses and outsider cell lines. As an available and reliable method for monitoring the purity of the cell cultures, we offer to use PCR kits designed and officially used in clinical diagnostics. We have tested 50 human cell lines using commercial diagnostic systems for detection of papilloma viruses, herpes viruses, adenoviruses, Mycoplasma hominis and total bacterial mass. Contamination in tested cell lines was not found. In the case of cell lines that contain integrated parts of viral genomes, the presence of the respective DNA sequences was confirmed. The proposed diagnostic systems can be effectively used to control the purity of cell lines, for qualitative detection of possible contamination, as well as for quantitative evaluations with calculation of viral load like it is practiced in clinical diagnostics.

The TRP (transient receptor potential) superfamily consists of a wide range of cation channels with different physiological functions and different cellular distribution. The TRP channels are implicated in pain perception, hearing and temperature sensation, photoreception, mechanotransduction, Ca2+-(re)absorption, cell proliferation and differentiation. Initially it was believed that TRP channels exist exclusively in the plasma membrane of the cells. Now this view is changing, since many members of this superfamily were found in the intracellular organelles where they successfully fulfill their conductive function. This review presents the current data concerning the members of the superfamily of TRP, which are integrated into the intracellular compartments. The role of TRP channels in the complicated and multicomponent machinery of endosomal pathways is analyzed.

Cultivation of cells under artificial conditions is one of the necessary procedures when working with stem cells. At this stage, the cells lose control by the macroorganism, becoming independent systems. Therefore, during the passaging of the cells, the probability of undesirable modifications is increased. At present, there is not enough data on the first indications about modification of the cultured cells and when they can appear. In the work, the rabbit medullary mesenchymal stromal cells (MSCs) were studied during 5 passages after producing primary culture. The area and the spreading coefficient of a cell were used to evaluate the cell state. The measurements were made 1 h and 1 day after cell reseeding by analyzing digital images of intact living cells. During this time interval, the absolute values of the cell area increased with the passage number, whereas the increment of the cell growth area did not depend on the passage number. It is possible to distinguish three groups of cells. Smaller cells (the 1st group) prevail in cell population of the 1st passage. Next, their proportion is decreased and the proportion of larger cells is increased. Passaging of the cells does not influence their spreading coefficient.

Diatoms are unicellular microscopic eukaryotes able to form species-specific frustules with micro- and nanostructures of biogenic silica. The morphogenesis of frustules components occurs intracellularly within special organelles under control of cytoskeleton. The mechanisms of morphogenesis control are not clear. The influence of paclitaxel, the microtubule inhibitor, onto morphogenesis of valve of pennate diatom Synedra acus subsp. radians was investigated. It has been shown that microtubule disturbance on the early morphogenesis stages provoke symmetry axis displacement of immature valve followed by curvature of the valve. At subsequent stages of morphogenesis stages, disturbance of the fine silica micro- and submicroscale structures (areolae and velums) occurs. It is supposed that a necessary condition for successful diatom valve morphogenesis is a balance between stable and dynamic microtubules.

The article presents the results of the studies of cytotoxic effect of zearalenone and T-2 toxin on sperm of horses and bulls during incubation and after thawing according to the technology of sperm obtaining and cryopreservation in Kharkov. We first have shown in vitro toxic effects of different concentrations of zearalenone and T-2 toxin (from 0.5 to 0.01 mM) on the membrane stability, as well as quantitative and qualitative indicators of semen in stallions and bulls before and after freezing and thawing. It has been found that the biological activity of the native sperm in 1 h after addition of 0.5 mM zearalenone is not changed, and after thawing is reduced by 19.4 %, or 0.69 ball (P < 0.01) compared to control without toxin. T-2 toxin at a concentration of 0.5 mM reduced the activity of the native sperm after an hour of incubation by 0.59 ball (P < 0.01) and decreased it after thawing decreased by 60.28 %, or 2.14 ball (P < 0.001). 1 h of incubation with zearalenone at concentration of 0.25 mM did not affect the activity of the native sperm, but the activity after freeze—thaw deteriorated by 12.2 %, or 0.41 ball (P < 0.05). T-2 toxin at a concentration of 0.25 mM reduced activity of native sperm after 1 h exposure by 0.46 ball (P < 0.05), and after thawing spermatozoa degrade it by 1.77 ball (P < 0.001) compared to control without toxins. In our study, the lowest concentration giving a significant decrease in sperm parameters was 0.01 mM. The negative impact of zearalenone in such a dose on the native sperm was absent, and sperm after thawing only slightly reduced activity and perezhivayemost by 0.09 points and 0.27 hours, respectively. The simultaneous addition of zearalenone and T-2 toxin at the same dose of 0.01 mM resulted in a decrease in biological perezhivayemost and absolute indicator of native sperm perezhivayemost by 0.36 points and 0.64 hours (P < 0.05), respectively.

We have investigated the organization of microtubule system in interphase cells of Medicago sativa L. roots during acclimation to salt and osmotic stress at different concentrations of NaCl, Na2SO4 and mannitol. We have identified several morphological changes in tubulin cytoskeleton that appear during the acclimation to salt and osmotic stress in the cells of different root tissues: 1) decreased density of cortical microtubule network, 2) random orientation of cortical microtubule bundles, 3) non-uniform density of the bundles, 4) thickening of the bundles, 5) fragmentation of the bundles, 6) formation of centers of converging microtubule. Reduced density of the microtubule network and thickening of the bundles was detected during osmotic and salt stress, yet random orientation of cortical microtubules was observed under osmotic stress and not found during salt stress. Fragmentation of microtubule bundles was apparent during salt stress and less evident at high concentration of mannitol. Formation of centers of converging microtubule was common under prolonged action of sodium sulfate, less common under sodium chloride and not found after mannitol treatment. Our data show that cortical microtubules in alfalfa root cells rearrange not only in response to different ions, but also to osmotic pressure. Thus, the signaling pathways and molecular mechanisms inducing reorganization of the microtubule system may be triggered not only by sodium cations but also by sulfate and chloride anion at the concentrations that do not cause irreversible cell damage. Our study show that the osmotic and salt stress differently affect the cortical microtubules, and their reorganization in response to stress depends on the salt cations as well as anions might also show additional effect under salt stress.

The adipose tissue is considered today as an endocrine organ in which tissue-specific regulation of gene expression plays a key role in the processes of development of obesity and comorbidities, such as diabetes and cardiovascular disease. The present study is focused on ITLN1, PPARã and TNFá gene expression in intra-abdominal adipose tissue and its effect on the serum levels of omentin 1 and TNFa in individuals with different body mass. It has been shown that serum TNFa level is significantly higher in the subgroup of patients with overweight and obesity (BMI ≥ 25 kg/m2) as compared to individuals with normal body weight (BMI < 25 kg/m2)(p < 0.03). We have demonstrated that the expression level of the PPARã gene is positively correlated with the ITLN1 gene expression level in the intra-abdominal adipose tissue (r = 0.516, p = 0.020). Serum level of omentin 1 positively correlates with PPARã mRNA and protein levels in intra-abdominal adipose tissue (r = 0.550, p < 0.05 and r = 0.581, p < 0.03, respectively). For the subgroup of patients with overweight and obesity, we have shown negative correlation of the level of TNFá mRNA with PPARã and ITLN1 mRNA levels was shown (r = –0.549, p < 0.05 and r = –0.475, p < 0.05, respectively). This study is the first to show a correlation relationship between PPARã gene expression level in the intra-abdominal adipose tissue and the expression and secretion levels of omentin 1.

Immunohistochemical analysis was used to study depigmented skin areas such as macular of depigmentation and skin perimakular areas in vitiligo patients. It has been shown that the cells containing melanocytic cell marker TRP1 are localized both in macular and perimakular areas. Within the macula of depigmentation all TRP1 positive cells are in close contact with the basement membrane. In perimakular areas many cells that have lost contact with the basement membrane, were localized deep in the epidermis. About 92 % of TRP1 positive perimakular cells were also vimentin positive. Vimentin positive cells were numerous in perimakular areas but missing in the macula of depigmentation. Dense groups of cells immunopositive for transcription factor Snail, known as inductor of epithelial-mesenchymal transition, were localized in perimakular areas in close proximity to the macula depigmentation border. Such cells were extremely rare within the macula of depigmentation. There is reason to assume that an intensive process, which is similar to the epithelial-mesenchymal transition, might be the cause of melanocyte death in perimakular field, and thus prevents repigmentation of depigmented areas.