Prions are alternative infectious conformations for some cellular proteins. For the protein PrP^C (PrP – prion protein, С – common), a prion conformation, called PrP^Sc (S – scrapie), is pathological. For example, in mammals the PrP^Sc prion causes transmissible spongiform encephalopathies accumulating in the brain tissues of PrP^Sc aggregates that have amyloid properties. MicroRNAs and long non-coding RNAs can be translated into functional peptides. These peptides can have a regulatory effect on genes from which their non-coding RNAs are transcribed. It has been assumed that prions, like peptides, due to the presence of specific domains, can also activate certain non-coding RNAs. Some of the activated non-coding RNAs can catalyze the formation of new prions from normal protein, playing their role in the pathogenesis of prion diseases. Confirmation of this assumption is the presence of the association of alleles of microRNA with the development of the disease, which indicates the role of the specific sequences of noncoding RNAs in the catalysis of prion formation. In the brain tissues of patients with prion diseases, as well as in exosomes containing an abnormal PrP^Sc isoform, changes in the levels of microRNA have been observed. A possible cause is the interaction of the spatial domains of PrP^Sc with the sequences of the non-coding RNA genes, which causes a change in their expression. MicroRNAs, in turn, affect the synthesis of long non-coding RNAs. We hypothesize that long noncoding RNAs and possibly microRNAs can interact with PrP^C catalyzing its transformation into PrP^Sc. As a result, the number of PrP^Sc increases exponentially. In the brain of animals and humans, transposon activity has been observed, which has a regulatory effect on the differentiation of neuronal stem cells. Transposons form the basis of domain structures of long non-coding RNAs. In addition, they are important sources of microRNA. Since prion diseases can arise as sporadic and hereditary cases, and hereditary predisposition is important for the development of pathology, we hypothesize the role of individual features of activation of transposons in the pathogenesis of prion diseases. The activation of transposons in the brain at certain stages of development, as well as under the influence of stress, is reflected in the peculiarities of expression of specific non-coding RNAs that are capable of catalyzing the transition of the PrP^C protein to PrP^Sc. Research in this direction can be the basis for targeted anti-microRNA therapy of prion diseases.

Matrix metalloproteinases (MMPs) are important for the pathogenesis of psoriasis and other autoimmune disorders. In the extracellular matrix, accumulation of proinflammatory cytokines, such as interleukin 17A (IL-17A), leads to induction of several MMPs, including MMP1. MMPs change the composition and other properties of the extracellular matrix. These changes facilitate tissue remodeling and promote the development of psoriatic plaques. The aim of this study was to explore how MMP1 silencing might influence the biological effects of IL-17A on migration and proliferation of human epidermal keratinocytes and the expression of genes involved in their division and differentiation. The experiments were performed with MMP1-deficient and control epidermal keratinocytes, HaCaT-MMP1 and HaCaT-KTR, respectively. Cell proliferation and migration were assessed by comparative analysis of the growth curves and scratch assay, respectively. To quantify cell migration, representative areas of cell cultures were photographed at the indicated time points and compared to each other. Changes in gene expression were analyzed by real-time PCR. The obtained results demonstrated that MMP1 silencing in the cells treated with IL-17A resulted in downregulation of MMP9 and -12, FOSL1, CCNA2, IVL, KRT14 and -17 as well as upregulation of MMP2, CCND1 and LOR. Moreover, MMP1 silencing led to a decrease in cell proliferation and an impairment of cell migration. Thus, MMP1-deficiency in epidermal keratinocytes can be beneficial for psoriasis patients that experience an accumulation of IL-17 in lesional skin. Knocking MMP1 down could influence migration and proliferation of epidermal keratinocytes in vivo, as well as help to control the expression of MMP1, -2, -9 и -12, CCNA2, CCND1, KRT14 and -17 that are crucial for the pathogenesis of psoriasis.

Endothelial keratoplasty has become the treatment of choice for corneal endothelial dysfunction. Advancements in the surgical treatment of corneal endothelial diseases depend on progress in graft conservation and its related advantages in assessing the suitability of grafts for transplantation. Transport of water and ions by cornea endothelium is important for the optic properties of cornea. In this work, we study the intracellular sodium concentration in cornea endothelial cells in samples of pig cornea that underwent hypothermic conservation for 1 and 10 days and endothelial cells of human cornea grafts after 10-day conservation. The concentration of intracellular sodium in preparations of endothelial cells was assayed using fluorescent dye SodiumGreen. The fluorescent images were analyzed with the custom-made computer program CytoDynamics. An increased level of intracellular sodium was shown in the endothelium after 10-day conservation in comparison with one-day conservation (pig samples). Sodium permeability of pig endothelial cell plasma membranes significantly decreased in these samples. Assessment of intracellular sodium in human cornea endothelium showed a higher level – as was in analogues pig samples of the corneal endothelium. The assay of the intracellular sodium balance concentration established in endothelial cells after hypothermic conservation in mediums L-15 and Optisol-GS showed a significant advantage of specialized me dium Optisol-GS. The balanced intracellular concentration after 10 days of hypothermic conservation was significantly lower in cells incubated at 4 °C in Optisol-GS (L-15, 128 ± 14,  n = 15; Optisol-GS, 108 ± 14, n = 11; mM, p < 0.001). Intracellular sodium concentration could be a useful parameter for assessing cornea endothelium cell viability.

Transcription factor gene ZFHX3 is one of the candidate genes involved in stroke development. The ZFHX3 protein contains oligopeptides encoded by trinucleotide repeats (TNRs). TNR variability is considered to be one of the causes of the disease, but their biological function has not yet been established. We assume that TNRs are the binding sites of miRNA to mRNA and are involved in regulation of ZFHX3 gene expression. The characteristics of miRNA–mRNA interaction were determined using MirTarget software. It has been shown that the first TNR in mRNA of the human ZFHX3 gene consists of the seven consecutive miR-12-32603-3p binding encoding polyGlu. The ZFHX3 protein of human polyGlu contains 30 Glu. In the orthologous proteins of 36 animal species the length of polyGlu varied from 27 Glu to 33 Glu. Negatively charged polyGlu of the ZFHX3 transcription factor probably interacted with positive DNA-binding proteins. The following mRNA region of the ZFHX3 gene contained the binding sites for miR-17-39416-3p, miR-5-15733-3p, miR-9-20317-3 encoding polyAla by 15 Ala lengths. In the 33 ZFHX3 orthologous proteins polyAla had the same length. The mRNA region of the human ZFHX3 gene with binding polysite of miR-1322-3p encoded polyGln consisting of 19 Gln. In the 41 orthologs of the ZFHX3 protein the length of polyGln varied from seven Gln to 23 Gln. The binding sites of miR-2-6184-3p, miR-5-14114-5p and miR-19-43437-5p were located with overlapping nucleotides sequences, and encode polyPro. In ZFHX3 human polyPro consisted of 12 Pro. In the orthologs, polyPro contained from 10 Pro to 14 Pro. The binding sites of miR-17-39416-3p, miR-9-20317-3p, miR-1-1819-3p, miR-5-15733-3p, miR-6-17815-3p, miR-18-39953-5p, miR-26862-5p, miR-1260b and miR-X-48174-3p in human ZFHX3 encoded polyGly by 22 Gly length. In the 28 orthologs of ZFHX3 the length of polyGly decreased to 11 Gly. The TNR regions could simultaneously bind several miRNAs, which increased the dependence of gene expression on miRNA. The oligopeptides encoded by the binding polysites of miRNA in mRNA in the orthologous ZFHX3 proteins were flanked by conserved oligopeptides.

Catatonia is a psychopathological syndrome displayed as a motor disorder. Catatonia is a sign of many menthal disorders, particularly schizophrenia and depression, with a wide disrtibution in the human population. The GC (“genetic” and “catatonia”) rat strain was obtained from the Wistar rat strain by a long selection (78 generations) for the catatonic type of reaction and is a model of schizophrenic and depressive disorders in humans. It is known that selection for behavior including catatonic reactions results in neuroendocrine, reproductive and morphological changes in animals. However, the influence of selection for a catatonic reaction on the spermatogenic function of testes had not been studied. The aim of this study was to conduct a comparative investigation of sperm quality in rats of the GC and the Wistar strain. The epididymal sperm parameters (sperm count, sperm motility, sperm morphology) were measured, and body, testes and epididymal weight were determined at puberty (50 day of life) and at adulthood (90 day of life). The litter size of the GC and Wistar rats was determined. It was found that adult GC rats had a lower sperm count, sperm motility, testis weight, epydidymal weight and litter size compared to adult Wistar rats. However, at puberty, GC rats had a higher sperm count than the Wistar strain. Interstrain differences in sperm morphology were not found. It has been assumed that the changes of spermatogenic parameters in response to selection for catatonia are caused by changing the ontogenic pattern of testosterone secretion. In conclusion, the hereditary predisposition to catatonic reaction is associated with impaired sperm parameters in adult rats that reduces their chance to reproduction. The GC rat strain can be a perspective model for investigation of the relationship between the hereditary predisposition to catatonia and spermatogenesis

Obesity during pregnancy increases the risk of obesity in offspring. To correct the offspring development in obese mothers, it is necessary to reveal the molecular mechanisms that mediate the influence of the maternal environment on the offspring ontogenesis. Leptin levels increase with obesity. In C57Bl mice, the А^у mutation is associated with elevated blood levels of leptin in pregnant females and exerts a gender-specific effect on the metabolic phenotype of mature offspring. Aim: to study the influence of А^у mutation on sensitivity to diet-induced obesity in male and female offspring, on fetal and placental weight and on the expression of genes in the placentas of the fetuses of different sexes. Body weight and food intake on a standard and an obesogenic diet, fetal and placental weights on pregnancy days 13 and 18, and gene expression of glucose transporters (GLUT1, GLUT3), neutral amino acid transporters (SNAT1, SNAT2, SNAT4), insulin-like growth factor 2 IGF2 and its receptor IGF2R were measured in male and female offspring of и ɑ/ɑ (control) and А^у/ɑ mothers. A^y mutation influenced the body weight only in male offspring, which consumed a standard diet, and did not influence obesity development in both male and female offspring. The weight of fetuses and placentas in А^у/ɑ as compared to ɑ/ɑ  females was reduced on day 13 of pregnancy and was not different on day 18. On day 13 of pregnancy, the mRNA levels of the examined genes did not differ in placentas of male and female fetuses in ɑ/ɑ  females. In А^у/ɑ females, the gene expression of GLUT1, GLUT3, SNAT1 and SNAT4 was reduced in female placentas compared to male placentas. The results suggest that the sex-specific transcription response of placentas to elevated leptin levels in pregnant А^у/ɑ females can mediate the gender-specific impact of А^у mutation on the offspring metabolism in postnatal life.

Vaccination forms active immunity and represents an effective way of preventing tick-borne encephalitis (TBE). However, excessive vaccination is unjustified in terms of economics and medical ethics. One of the individualized approaches to vaccines is the selection of vaccine doses depending on the expected levels of immune response. Therefore, there is a need for new methods for assessing potential human immune responses prior to vaccination. The aim of this study was to determine possible association between single nucleotide polymorphisms (SNPs) located within OAS2 and OAS3 genes, which have been previously associated with the development of severe forms of TBE, and the formation of antibodies and cytokines upon vaccination against TBE. The study involved 97 volunteers of both sexes who had not previously been vaccinated against TBE and had no contact with ticks. Venous blood samples were collected one month after vaccination against TBE using the EnceVir vaccine. Levels of specific IgG antibodies against tick-borne encephalitis virus and interleukin 4 (IL-4) were analyzed. Genomic DNA samples were genotyped for the SNPs rs2285932, rs2072136, rs1293762, rs15895 and rs1732778 in genes encoding 2’-5’-oligoadenylate synthetases OAS2 and OAS3. Antibody production in response to vaccine administration was significantly associated with SNP rs1732778 in the regulatory region of the OAS2 gene. This indicator was significantly higher in people with heterozygous genotypes G/A as compared to people with homozygous genotypes G/G and A/A. Carriers of the A allele (G/A or A/A genotypes) of the same SNP had reduced IL-4 levels as compared to the homozygous G/G individuals. Thus, the data obtained indicate that SNP rs1732778 in the regulatory region of the OAS2 gene correlates with the formation of antiviral IgG antibodies and changes in IL-4 levels upon vaccination. Evidently, the genetic polymorphism in OAS2 gene should be considered when performing individualized TBE vaccinations.

In selecting rats for behavior, we observe a direct natural effect and affect the nonspecific stress function. In this process, new behavioral phenotypes appear in the strain under selection. They differ from the selected forms in the selection criterion. In the GC strain, a large proportion of the so-called nervous rats emerge. The criterion presumes the selection for the long cataleptic freezing character, whereas the nervous rats display elevated motor excitement: running, jumping, and vocalization. The main purpose of our study was to assess phenotypic indices in GC rats (abbreviated from genetic and catatonia) and recognize principal components of variability for emotional and weight indices. Rats of the ancestral Wistar population were taken as control. The following indices were measured: time of cataleptic freezing, excitement level, blood pressure, acoustic startle response, seizure  activity, and weights of the heart, kidneys, adrenals, and spleen. Multivariate analysis methods were applied: factor analysis and principal component analysis. We confirmed the inclination of GC rats of the generation studied to freezing in quiet surrounding and after a strong acoustic sti - mulus. More pronounced startle responses,  moderate hypertension, and larger weights of the heart and adrenals were noted. Two principal variability components were recognized: startle amplitude (PC1) and morphofunctional variability (PC2). The figure shows different locations of Wistar and GC individuals in principal component coordinates. The principal component method confirmed the genetic relationship between the startle and nervousness responses. It was shown that in PC2 the indices of heart, kidney, adrenal, and spleen weight exert negative effects, whereas the effects of startle and nervousness were positive. In the same component, an increase in the startle and nervousness responses positively correlates with the relative weights of the heart and adrenals. Differences in the directions of the contributions to the second component of morphofunctional variability are discussed.

A drug for the prevention and therapy of tick-borne encephalitis virus is being developed on the basis of the protective chimeric antibody ch14D5a. At the same time, the epitope recognized by this antibody on the surface of glycoprotein E has not been localized yet. The aim of this work was to identify the domain of glycoprotein E, to which the protective antibody ch14D5a binds. As a result, four recombinant variants of glycoprotein E were generated using the bacterial expression system: (1) the rE protein containing the domains D1, D2, and D3 of glycoprotein E; (2) the rED1+2 protein containing domains D1 and D2; (3) the rED3_301 protein, which is domain D3 of glycoprotein E, and (4) the rED3_294 protein comprising domain D3 and a hinge region connecting domains D1 and D3. The rED3_294 and rED3_301 proteins were obtained in soluble monomeric form. The rE and rED1+2 proteins were extracted from the inclusion bodies of Escherichia coli. Using Western blot analysis and surface plasmon resonance analysis, it was demonstrated that the protective chimeric antibody ch14D5a and its Fab fragment bound specifically to domain D3 of glycoprotein E. Since the antibodies recognizing epitopes on the surface of domain D3 do not tend to cause antibody-dependent enhancement of the infection as compared to antibodies directed to domains D1 and D2, the data obtained confirm the promise of using the antibody ch14D5a in the development of a therapeutic preparation against the tick-borne encephalitis virus.

The wide spread of hybridization in the genus Populus, including spontaneous hybridization, caused by cultivars, requires studying the variability and inheritance of morphological traits by hybrids for initial tracking of these processes. The analysis of endogenous, intra- and inter-population variability was performed on 533 individual poplar trees in seven populations of P. nigra, seven populations of P. laurifolia and four populations of P. × jrtyschensis in the Tom river basin. On each specimen, 15 leaves from short mid-crown branches were collected to determine the shape of the leaf blade, the shape of its tip and base, as well as the branch morphotype. Some biometric indicators were proposed for geometric assessment of the leaf blade shapes of poplar species. The analysis showed that of all the traits examined only the leaf blade shape did not meet the criterion for “phene”, since it is usually represented by several forms in the crown of one and the same tree. In all the species studied, the level of their intra-population diversity was found to be much higher than the inter-population one. According to the increase of intra-population variability of qualitative traits, the taxa could be ranked as P. nigra < P. laurifolia < P. × jrtyschensis. The share of inter-population diversity differed among the species studied, accounting for 21.5% in P. laurifolia, 13.8% in P. nigra and 8.0% in P. × jrtyschensis. The P. laurifolia populations showed the greatest inter-population differentiation, most likely because of a disjunct distribution due to narrow specialization to the montane river environment. The lower differentiation in P. nigra is probably due to the facts that this species dominates the poplar stands of the Tom River basin; its populations are not fragmented and are linked by vast gene flow. In P. nigra, an increase in the diversity of qualitative characteristics and phenotypes was observed in populations confined to hybridization centers. Natural selection is most likely the factor governing the inter-population differentiation in P. × jrtyschensis, leading to the predominance of F1 hybrids in populations and hence to a sharp decrease in inter-population variability.

Many garden chrysanthemums bred across the world are not fully winter hardy. Many are damaged by fungal diseases due to a high humidity and are late flowering. This makes them unsuitable for general commercial marketing. Since 2000 we have been conducting a breeding program using natural species of the genus Chrysanthemum that are adapted to the local conditions. The strategy of breeding adaptive hybrids and varieties of chrysanthemum native to Russia was proposed based on their biological, genetic peculiarities and natural resources of Chrysanthemum with the use of interspecific hybridization. Research objects are the first generations of interspecific hyb rids of F₁ obtained previously by the author as a result of the hybridization of varieties and wild species of Chrysanthemum. Derived from different species and varieties, F₁ hybrids were crossed among themselves to obtain the multicomponent F₂ progeny. F₂ seedlings with winter hardiness, resistance to Puccinia horiana Henn. and early flowering were used in closely related crosses. The offspring of F₃ from closely related crosses were also assessed and selected according to adaptive and decorative characteristics. Inclusion in the selection process of various sources of winter hardiness and resistance to P. horiana allowed positive characteristics to be increased in F₂ and to be revealed in F₃. Adaptive signs of the wild species Chrysanthemum naktongense Nakai, C. coreanum (H. Lév. et Vaniot) Nakai, C. zawadzkii var. tenuisectum Kitag., C. leiophyllum Nakai, and C. zawadzkii subsp. acutilobum (DC.) Kitag., which have formed and fixed during evolution, were inherited and manifested in offspring of the multicomponent hybrids and the closely related crosses. Promising interspecific forms with biological signs determining the possibility of growing in extreme conditions of the subregion were selected.

Cereal Crop Wild Relatives (CWR) are a very  important gene pool for cereal/wheat improvement. New genes for resistance to diseases and pests are urgently needed to avoid using pesticides and to raise adaptivity to the environmental stresses caused by global climate change. In this regard, the study is aimed at ex situ conservation of Aegilops L. genus local ecotypes’ genetic diversity, which is very relevant and promising for breeding. In order to establish breeding utility and form an ex situ collection reflecting the intra- and inter-specific diversity, the phenotypic screening of Kazakhstan’s local populations of Aegilops L. genus (Ae. cylindrica, Ae. tauschii, Ae. triuncialis and Ae. crassa) was conducted on the basis of multiple indicators. For the first time molecular-genetic analysis of 50 representatives of Aegilops L. genus from Kazakhstan’s flora was performed. The microsatellite analysis with the use of 11 EST-SSR markers revealed eight of them to be most effective. For each marker, allele frequency and average heterozygosity was calculated. For the most informative markers the presence of 5 and 6 respective allelic variations was found. A bank of genomic DNA was created and kept in ex situ storage (–70 °С, long-term) in the IMBB of the MES of RK.