Cognitive impairment in long-living adults: a genome-wide association study, polygenic risk score model and molecular modeling of the APOE protein.

Background: Cognitive impairment is an irreversible, aging-associated condition that robs people of their independence. The purpose of this study was to investigate possible causes of this condition and propose preventive options. Methods: We assessed cognitive status in long-living adults aged 90+ (n = 2,559) and performed a genome wide association study using two sets of variables: Mini-Mental State Examination scores as a continuous variable (linear regression) and cognitive status as a binary variable (> 24, no cognitive impairment; <10, impairment) (logistic regression). Results: Both variations yielded the same polymorphisms, including a well-known marker of dementia, rs429358in the APOE gene. Molecular dynamics simulations showed that this polymorphism leads to changes in the structure of alpha helices and the mobility of the lipid-binding domain in the APOE protein. Conclusion: These changes, along with higher LDL and total cholesterol levels, could be the mechanism underlying the development of cognitive impairment in older adults. However, this polymorphism is not the only determining factor in cognitive impairment. The polygenic risk score model included 45 polymorphisms (ROC AUC 69%), further confirming the multifactorial nature of this condition. Our findings, particularly the results of PRS modeling, could contribute to the development of early detection strategies for predisposition to cognitive impairment in older adults.

Features of the use of ladder myoplasty of a gunshot wound to the laryngopharynx: Case report.

INTRODUCTION AND IMPORTANCE: The laryngopharynx wound is considered to be one of the most severe wounds of neck both in war and in peace, as it may cause life threatening changes in the whole body (asphyxia, bleeding, shock). Important aspects of surgical treatment are to ensure full breathing, acceptable ways of feeding, and the use of reliable wound closure techniques aimed to prevent digestive tract failure and to maintain the framework and aerostasis of the laryngotracheal region. CASE PRESENTATION: A case of unilateral multiple wounds of the laryngopharynx was described in the article. The features of diagnostics, surgical treatment and conservative therapy in the postoperative period with this injury were presented. The wounded man was urgently operated. During surgery the pharynx was mobilized. The metal fragment was removed. The wound of the pharynx was sutured with a two-row suture. The next stage of the surgical treatment was myoplasty. In the case of the patient, the purpose of myoplasty was additional sealing of the pharyngeal suture and myoplasty of the thyroid cartilage injury zone for the purpose of aerostasis. Because of the size of the wounds and their anatomical localization, we used the mobilized lower edge of the Musculus sternocleidomastoideus for myoplasty and proposed the method of ladder myoplasty developed by us. CLINICAL DISCUSSION: In myoplasty method the following criteria must be followed: the muscle flap must be of sufficient length and width, so as not to cause excessive tension in the myoplasty area; the flap must be thick enough to avoid necrosis that may cause subsequent infectious complications; when taking the flap, the most sparing operative access should be used to avoid functional and anatomical disorders; the volume of the taken muscle flap must not lead to functional and anatomical disorders. CONCLUSION: The proposed method of ladder myoplasty using Musculus sternocleidomastoideus is unique, and proves its high efficiency in unilateral multiple laryngopharyngeal injuries, and can be recommended for wide clinical implementation in such clinical situations.

[The study of tryptophan metabolite concentrations in blood serum and fecal extracts from obese children]. / Izuchenie soderzhaniia metabolitov obmena triptofana v syvorotke krovi i kalovykh ékstraktakh u detei s ozhireniem.

We found that changes in the concentrations of tryptophan metabolites in the blood serum and in the intestinal contents are one of the mechanisms for the formation of metabolic coupling in the system "macroorganism-intestinal microbiota", which undergoes significant changes in the development of obesity. Although blood kynurenine remained basically unchanged in obese children we found an increase in some of its serum metabolites: anthranilic, kynurenic and xanthurenic acids. It is noteworthy that in the analysis of fecal matter in obese children, revealed a 2-fold increase in the level of kynurenine while the concentration of kynurenine pathway metabolites corresponded to the level of the group of healthy children. This may indicate the metabolic activation of the microbiota associated with the intestinal mucosa. This is also supported by the absence of statistically significant differences in the concentration of indole in healthy children and in obese children in fecal analyses, and a significant increase in the concentration of indole-3-lactate and indole-3-acetate in the blood serum of obese children.

Functional Analysis of Coilin in Virus Resistance and Stress Tolerance of Potato Solanum tuberosum using CRISPR-Cas9 Editing.

The role of the nuclear protein coilin in the mechanisms of resistance of potato Solanum tuberosum cultivar Chicago to biotic and abiotic stresses was studied using the CRISPR-Cas9 technology. For the coilin gene editing, a complex consisting of the Cas9 endonuclease and a short guide RNA was immobilized on gold or chitosan microparticles and delivered into apical meristem cells by bioballistics or vacuum infiltration methods, respectively. Editing at least one allele of the coilin gene considerably increased the resistance of the edited lines to infection with the potato virus Y and their tolerance to salt and osmotic stress.

Guide RNA Design for CRISPR/Cas9-Mediated Potato Genome Editing.

The activity of the pool of sgRNA molecules designed for different regions of potato coilin and phytoene desaturase genes was compared in vitro. Due to the presence of nucleotides unpaired with DNA, sgRNA is able not only to inhibit but also to stimulate the activity of the Cas9-sgRNA complex in vitro. Although the first six nucleotides located in the DNA substrate proximally to the PAM site at the 3' end are the binding sites for cas9, they had no significant effect on the activity of the Cas9-sgRNA complex.

Non-structural Functions of Hordeivirus Capsid Protein Identified in Plants Infected by a Chimeric Tobamovirus.

Capsid proteins (CPs) of (+)RNA-containing plant viruses are multifunctional proteins involved in many stages of viral infection cycle, in addition to their main function of virus capsid formation. For example, the tobamoviral CP ensures virus systemic transport in plants and defines the virus-host interactions, thereby influencing the virus host range, virus infectivity, pathogenicity, and manifestation of infection symptoms. Hordeiviruses and tobamoviruses belong to the Virgaviridae family and have rod-shaped virions with a helical symmetry; their CPs are similar in structure. However, no non-structural functions of hordeiviral CPs have been described so far. In this study, we assayed possible non-structural functions of CP from the barley stripe mosaic virus (BSMV) (hordeivirus). To do this, the genome of turnip vein clearing virus (TVCV) (tobamovirus) was modified by substituting the TVCV CP gene with the BSMV CP gene or its mutants. We found that BSMV CP efficiently replaced TVCV CP at all stages of viral infection. In particular, BSMV CP performed the role of tobamoviral CP in the long-distance transport of the chimeric virus, acted as a hypersensitive response elicitor, and served as a pathogenicity determinant that influenced the symptoms of the viral infection. The chimeric tobamovirus coding for the C-terminally truncated BSMV CP displayed an increased infectivity and was transported in plants in a form of atypical virions (ribonucleoprotein complexes).

FEATURES OF EPISOTIC PROCESS AND MOLECULAR-GENETIC CHARACTERISTICS OF VIRUS ISOLATES OF RABIES IN TVER REGION.

Rabies epidemic situation in the Tver Region has been studied. Animals of different species that had confirmed clinical rabies were statistically analyzed. It was established that the features of the course of epizootics in the Tver region correspond to the regularities characteristic of rabies of the natural-focal type. As a result of sequencing of the rabies virus N gene and phylogenetic analysis, the isolates studied were assigned to the central phylogenetic group. With the help of the geoinformatic system, nosological maps of the Tver region were obtained and the spatial- temporal features of the course of the epizootic process of rabies infection were studied.

Searching for active mobile genetic elements in dsRNA fraction of Pinus sylvestris having witches broom abnormalities.

The most common type of coniferous mobile genetic elements are retrotransposons. Despite of their early positive impact on evolution of modern coniferous species they can have a significant negative impact for Forestry and breeding. Breaking genomic structural integrity mobile elements can cause phenotypic defects of plants. In this regard, the study of the diversity of coniferous mobile genetic elements is particularly interesting. In the present paper, we describe mobile genetic elements in dsRNA fraction of Pinus sylvestris having witches broom abnormalities. In result of assembled contigs analysis by RepeatMasker 70 mobile genetic elements were identified. A 68 of that were retroelements. Most of elements represented by Gypsy (16 contigs) and Copia (48 contigs). In 4 cases retroelements specific to Pinus taeda were identified. In most cases fragments of integrase (24), reverse transcriptase (22) and RNaseH (15) were identified. Results of the study may be of interest for coniferous breeding and genetic specialists. The raw data of these experiments have been deposited at NCBI under the accession number SAMN06185845.

[ZOOPATHOGENIC ORTHOBUNIAVIRUSES (ORTHOBUNYAVIRUS, BUNYAVIRIDAE)].

This work deals with the systematics and taxonomy of orthobunyaviruses, little-studied dangerous and new iruses Akabane, Aino, Schmallenberg, Cache Valley diseases, Oropouche fever. The significance of the reassortment mechanism of their origin and diversification is discussed.

Helical capsids of plant viruses: architecture with structural lability.

Capsids of numerous filamentous and rod-shaped plant viruses possess helical symmetry. In positive-stranded RNA viruses, helical capsids are typically composed of many identical subunits of the viral capsid protein (CP), encapsidating a molecule of viral genomic RNA. Current progress in structural studies of helical plant viruses has revealed differences between filamentous and rod-shaped viruses, both in structural folds of their CPs and in the interactions of CP molecules in their capsids. Many filamentous and rod-shaped viruses have functionally similar lateral inter-subunit contacts on the outer virion surface. Additionally, the extreme N-terminal CP region in filamentous viruses is intrinsically disordered. Taken together, the available data establish a link between the structural features of molecular interactions of CP molecules and the physical properties of helical virions ranging from rigidity to flexibility. Overall, the structure of helical plant viruses is significantly more labile than previously thought, often allowing structural transitions, remodelling and the existence of alternative structural forms of virions. These properties of virions are believed to be functionally significant at certain stages of the viral life cycle, such as during translational activation and cell-to-cell transport. In this review, we discuss structural and functional features of filamentous and rod-shaped virions, highlight their shared features and differences, and lay emphasis on the relationships between the molecular structure of viral capsids and their properties including virion shape, lability and capability of structural remodelling.

Structure and Noncanonical Activities of Coat Proteins of Helical Plant Viruses.

The main function of virus coat protein is formation of the capsid that protects the virus genome against degradation. However, besides the structural function, coat proteins have many additional important activities in the infection cycle of the virus and in the defense response of host plants to viral infection. This review focuses on noncanonical functions of coat proteins of helical RNA-containing plant viruses with positive genome polarity. Analysis of data on the structural organization of coat proteins of helical viruses has demonstrated that the presence of intrinsically disordered regions within the protein structure plays an important role in implementation of nonstructural functions and largely determines the multifunctionality of coat proteins.

[ROENTGENOLOGICAL IMAGING FEATURES IN PATIENTS WITH LUNG CANCER AND METASTATIC OF LUNG AFFECTION].

The most often radiological imaging features in patients with lung cancer and metastasis of lung were presented.

Structural Properties of Potexvirus Coat Proteins Detected by Optical Methods.

It has been shown by X-ray analysis that cores of coat proteins (CPs) from three potexviruses, flexible helical RNA-containing plant viruses, have similar α-helical structure. However, this similarity cannot explain structural lability of potexvirus virions, which is believed to determine their biological activity. Here, we used circular dichroism (CD) spectroscopy in the far UV region to compare optical properties of CPs from three potexviruses with the same morphology and similar structure. CPs from Alternanthera mosaic virus (AltMV), potato aucuba mosaic virus (PAMV), and potato virus X (PVX) have been studied in a free state and in virions. The CD spectrum of AltMV virions was similar to the previously obtained CD spectrum of papaya mosaic virus (PapMV) virions, but differed significantly from the CD spectrum of PAMV virions. The CD spectrum of PAMV virions resembled in its basic characteristics the CD spectrum of PVX virions characterized by molar ellipticity that is abnormally low for α-helical proteins. Homology modeling of the CP structures in AltMV, PAMV, and PVX virions was based on the known high-resolution structures of CPs from papaya mosaic virus and bamboo mosaic virus and confirmed that the structures of the CP cores in all three viruses were nearly identical. Comparison of amino acid sequences of different potexvirus CPs and prediction of unstructured regions in these proteins revealed a possible correlation between specific features in the virion CD spectra and the presence of disordered N-terminal segments in the CPs.

"Green" nanotechnologies: synthesis of metal nanoparticles using plants.

While metal nanoparticles are being increasingly used in many sectors of the economy, there is growing interest in the biological and environmental safety of their production. The main methods for nanoparticle production are chemical and physical approaches that are often costly and potentially harmful to the environment. The present review is devoted to the possibility of metal nanoparticle synthesis using plant extracts. This approach has been actively pursued in recent years as an alternative, efficient, inexpensive, and environmentally safe method for producing nanoparticles with specified properties. This review provides a detailed analysis of the various factors affecting the morphology, size, and yield of metal nanoparticles. The main focus is on the role of the natural plant biomolecules involved in the bioreduction of metal salts during the nanoparticle synthesis. Examples of effective use of exogenous biomatrices (peptides, proteins, and viral particles) to obtain nanoparticles in plant extracts are discussed.

Subterahertz chaos generation by coupling a superlattice to a linear resonator.

We investigate the effects of a linear resonator on the high-frequency dynamics of electrons in devices exhibiting negative differential conductance. We show that the resonator strongly affects both the dc and ac transport characteristics of the device, inducing quasiperiodic and high-frequency chaotic current oscillations. The theoretical findings are confirmed by experimental measurements of a GaAs/AlAs miniband semiconductor superlattice coupled to a linear microstrip resonator. Our results are applicable to other active solid state devices and provide a generic approach for developing modern chaos-based high-frequency technologies including broadband chaotic wireless communication and superfast random-number generation.

[Virosphere and giruses].

Novel findings and concepts in the field of virology particularly regarding virosphere and giruses--a group of large nuclear-cytoplasmic deoxyriboviruses are briefly summarized. In the context of novel understanding the major taxonomic features and virus pathogenicity including African swine plague are interpreted.

Subcellular localization and self-interaction of plant-specific Nt-4/1 protein.

The Nicotiana tabacum Nt-4/1 protein is a plant-specific protein of unknown function. Analysis of bacterially expressed Nt-4/1 protein in vitro revealed that the protein secondary structure is mostly alpha-helical and suggested that it could consist of three structural domains. Earlier studies of At-4/1, the Arabidopsis thaliana-encoded ortholog of Nt-4/1, demonstrated that GFP-fused At-4/1 was capable of polar localization in plant cells, association with plasmodesmata, and cell-to-cell transport. Together with the At-4/1 ability to interact with a plant virus movement protein, these data supported the hypothesis of the At-4/1 protein involvement in viral transport through plasmodesmata. Studies of the Nt-4/1-GFP fusion protein reported in this paper revealed that the protein was localized to cytoplasmic bodies, which were co-aligned with actin filaments and capable of actin-dependent intracellular movement. The Nt-4/1-GFP bodies, being non-membrane structures, were found in association with the plasma membrane, the tubular endoplasmic reticulum and endosome-like structures. Bimolecular fluorescence complementation experiments and inhibition of nuclear export showed that the Nt-4/1 protein was capable of nuclear-cytoplasmic transport. The nuclear export signal (NES) was identified in the Nt-4/1 protein by site-directed mutagenesis. The Nt-4/1 NES mutant was localized to the nucleoplasm forming spherical bodies. Immunogold labeling and electron microscopy of cytoplasmic Nt-4/1-containing bodies and nuclear structures containing the Nt-4/1 NES mutant revealed differences in their fine structure. In mammalian cells, Nt-4/1-GFP formed cytoplasmic spherical bodies similar to those found for the Nt-4/1 NES mutant in plant cell nuclei. Using dynamic laser light scattering and electron microscopy, the Nt-4/1 protein was found to form multimeric complexes in vitro.

In vitro phosphorylation of the N-terminal half of hordeivirus movement protein.

The N-terminal half of TGB1 movement protein of poa semilatent hordeivirus, which forms a ribonucleoprotein complex involved in movement of the viral genome in the plant, and its two domains, NTD and ID, are phosphorylated in vitro by a fraction enriched in cell walls from Nicotiana benthamiana. Using a set of protein kinase inhibitors with different specificities, it was found that enzymes possessing activities of casein kinase 1, protein kinase A, and protein kinase C are involved in phosphorylation. Commercial preparations of protein kinases A and C are able to phosphorylate in vitro recombinant proteins corresponding to the N-terminal half of the protein and its domains NTD and ID. Phosphorylation of the NTD has no effect on the efficiency and character of its binding to RNA. However, phosphorylation of the ID leads to a decrease in its RNA-binding activity and in the ability for homological protein-protein interactions.

The internal domain of hordeivirus movement protein TGB1 forms in vitro filamentous structures.

The 63 kDa hordeivirus movement protein TGB1 of poa semilatent virus (the PSLV TGB1 protein) forms viral ribonucleoprotein for virus transport within a plant. It was found using the dynamic laser light scattering technique that the internal domain of TGB1 protein forms in vitro high molecular weight complexes. According to results of atomic force microscopy, a part of these complexes is represented by globules of different sizes, while another part consists of extended filamentous structures. Similar properties are also characteristic of the N-terminal half of the protein and are obviously due to its internal domain moiety. The data support the hypothesis that upon viral ribonucleoprotein complex formation, the N-terminal half of the PSLV TGB1 protein plays a structural role and exhibits the ability to form multimeric filamentous structures (the ability for self-assembly).

[Mini-invasive operative interventions for spontaneous pneumothorax on the background of nonspecific pulmonary diseases].

The data, concerning the rate, causes and conditions of occurrence, localization of spontaneous pneumothorax, occurring on the background of non-specific pulmonary disease, tactic of its treatment, were adduced. The accent was done on the expediency of thoracoscopy performance for the correct diagnosis establishment as well as in the further tactic of treatment determination. The miniinvasive technologies introduction had permitted to improve the results of the patients treatment significantly and to prevent recurrence of spontaneous pneumothorax.