[Assessment of the phenotypic composition of lymphocytes, immunoglobulins and their relationships in patients with chronic rinosinusitis]. / Otsenka fenotipicheskogo sostava limfotsitov, immunoglobulinov i ikh vzaimootnoshenii u patsientov s khronicheskim rinosinusitom.

OBJECTIVE: The study of the population and subpopulation content of lymphocytes and immunoglobulins and their associations in IgE-mediated CRS relative to other CRS and the control group. MATERIAL AND METHODS: 23 patients with IgE-mediated chronic rhinusinusitis and 67 patients with normal IgE blood levels were examined. For analysis, flow cytometry (Cytomics FC500, Beckman Coulter, USA), using monoclonal antibodies CD3+, CD4+, CD8+, CD16+, CD19+ and enzyme immunoassay (Thermo Scientific Multiskan FC, Thermo Fisher Scientific, USA), using IgA, IgM, IgE and IgG in serum, statistical processing was performed using Statistica 7.0. RESULTS: In patients with CRS and IgE-mediated CRS, hyperactivation was revealed in both T- and B-cell immunity, manifested by an increase in the level of T-lymphocytes, NK-lymphocytes and B-lymphocytes. More pronounced disorders in the immune status are detected in patients with IgE - mediated CRS, there is a more pronounced activation of the T-cell immune link due to an increase in T-helper cells, T-killer/suppressor cells, an imbalance in their number is accompanied by a decrease in their ratio in the immunoregulatory index. Activation of the immune system in patients with CRS is also associated with an increase in the content of mature B-lymphocytes (CD19+), while only in patients with IgE-mediated CRS, hypergammaglobulinemia of classes A and M was detected. CONCLUSION: Changes in the immune status indicate a violation of immune regulation, confirmed by the revealed correlations between the subpopulations of lymphocytes and immunoglobulins that implement the immune response in this condition. The greatest number of violations in the regulation is associated with mature T-lymphocytes in both CRS and IgE-mediated CRS, while only IgA fully retains its function.

[DNA Fragment Enrichment for High-Throughput Sequencing].

This review describes the application of oligonucleotides, which are mainly obtained using DNA synthesizers of a new generation (microarray DNA synthesizers), for the enrichment of target genomic fragments. The methods of molecular hybridization, polymerase chain reaction, and CRISPR-Cas9 system for this purpose are considered. Examples of the practical use of the developed methods for research and diagnostic purposes are given.

[Social factors of mental health and well-being of older Russians.]

The article highlights social factors of mental health and well-being of older Russians based on the results of wave sociological studies of the perceived quality of life for 2002-2019 and the author's representative survey of older Russians in 2022. Age, gender and employment are the most significant determinants not only of social activity, but also of mental health and emotional well-being. At the same time, the study shows that the pandemic had a multidirectional impact on perceived quality of life among older Russians. Compared to the pre-pandemic situation, after the relaxation of restrictive measures, the parameter of social activity increased significantly, the parameter of mental health remained virtually unchanged, but the emotional problems of elderly Russians became more acute. Due to the severe restrictions for older people, the long period of isolation, and for working pensioners - also due to the need to restructure their work remotely - stress, anxiety and worry have affected everyday life.

[Effect of the elimination diet in psoriasis: a clinical case].

Psoriasis (PS) is a multifactorial disease with a dominant role of genetic predisposition, but the questions of PS etiology and pathogenesis still remain open. The development of PS can be facilitated by environmental factors, as well as a violation of the skin barrier and immune imbalance. In the literature of recent years, an association in clinical practice between PS and atopic diseases (atopic dermatitis, bronchial asthma, allergic rhinitis) has been increasingly reported. As a result of the increase in the prevalence of allergic diseases in the world, a special role is given to the study of nutrition, in particular food allergy in the development of PS. An elimination diet under food allergy is the main type of etiotropic therapy that prevents the launch of immunopathological inflammatory reactions. However, the literature does not provide data on the positive effect of the elimination effect in food allergy in patients with psoriasis. The purpose of this work was to present a clinical case of the effectiveness of the elimination diet for food allergy in a patient with PS. Methods. A specific allergological examination of a 65-year old patient with widespread PS (suffering from the age of 25) was carried out: determination of the concentration of total immunoglobulin E (IgE), eosinophilic cationic protein and allergen-specific IgE to food, pollen, fungal allergens in the blood serum by enzyme-linked immunosorbent assay, skin prick testing with food and pollen allergens. Taking into account the specific allergological examination, the patient was prescribed a diet with the elimination of causally significant allergens, including cross-reacting ones, for a period of 1-3 months (with the exception of chicken eggs, cereals, buckwheat, baker's yeast and products based on yeast fermentation, cereal products (bread, bakery, rolled oats, bran, oatmeal cookies, cereals, pasta); as well as peanuts, smoked sausages, coffee, cocoa, ice cream, sherbet, sesame, sorghum, honey and bee products, strawberries, wild strawberries, citrus, legumes, soybeans, sorrel, herbal teas, tree fruits (raw apples, peaches, cherries; nuts, exotic fruits), celery, raw carrots, tomatoes). Results. The demonstrated clinical case indicates that the appointment of an elimination diet in patients with PS with concomitant food allergies contributes to the rapid regression of the skin process. The increased concentration of total IgE in blood serum revealed during patient examination, the presence of positive reactions to food and pollen allergens according to the results of skin prick testing and the determination of allergen-specific IgE, the positive food elimination effect demonstrate the important role of allergic reactions in the development of skin lesions in PS. The presented observation demonstrates the importance of conducting a specific allergic examination in patients with PS, including an allergic history, determining the concentration of total and allergen-specific IgE, and eosinophilic cationic protein in blood serum, studying the spectrum of sensitization to food, pollen and fungal allergens by skin prick testing. Conclusion. Thus, as a result of the study, it was found that the appointment of an individual elimination diet, taking into account the results of a specific allergological examination, can contribute not only to the effective resolution of foci of psoriatic skin lesions, but also to the prevention of the progression of the systemic inflammatory process, reducing the risk of comorbid conditions and, therefore, improving the life quality of patients with psoriasis.

[Application of Array-Based Oligonucleotides for Synthesis of Genetic Designs].

The application of array-based oligonucleotides in biological studies is described. These oligonucleotides are mainly used to design large libraries of various nucleotide sequences, which are applied to study protein-nucleic acid interactions, splicing, transcription, translation, and other regulatory processes in mammalian, yeast, and bacterial systems. The application of gene libraries generated by array-based nucleotides along with advanced methods of the combination of DNA duplexes will make it possible to obtain complex genetic designs for synthetic biology.

Changes in the indices of prooxidant and antioxidant systems in blood plasma in men with atrophic gastritis and gastric cancer.

AIM: To study changes in the indices of prooxidant and antioxidant systems in plasma in men with atrophic gastritis and gastric cancer. MATERIALS AND METHODS: The study included 60 healthy men, 42 patients with atrophic gastritis and 50 men, nicardipine patients with gastric cancer stage II according to TNM. All patients underwent serological diagnosis of diffuse atrophic gastritis (definition of pepsinogens and gas- trin-17) and Helicobacter pylori infection. The diagnosis of "atrophic gastritis" was verified by morphological examination of biopsy speci- mens obtained during fibroesophagogastroduodenoscopy. Diagnosis of gastric cancer was carried out in the Krasnoyarsk regional oncologic dispensary on the basis of a comprehensive instrumental and morphological examination. All patients spectrophotometric methods in plasma was determined the content of diene conjugates (DC), malonic dialdehyde (MDA), glutathione-S-transferase (GST), glutathione peroxidase (GPO), superoxide dismutase (SOD) and catalase. RESULTS: The concentration of SOD, GST, GPO and catalase had no significant differences in patients with atrophic gastritis and gastric cancer and prevailed in comparison with healthy persons. Patients with cancer of the stomach content in the blood plasma DK 2.7 times and MDA at 35.2 times higher than healthy individuals, indicating severe oxidative stress in patients with cancer. In patients with atrophic gastritis was ob- served similar but less pronounced pattern. CONCLUSION: The results indicate the presence of oxidative stress in men with atrophic gastritis and gastric cancer.

[A prototype of oligonucleotide microarray for detection of pathogens relating to arena- and Filoviridae families].

A prototype of oligonucleotide microarray for detection of Lassa, Junin, Machupo, Guanarito viruses (Arenaviridae family), Ebola and Marburg viruses (Filoviridae family) was presented. An original approach founded on virus proteins (nucleocapsid protein for Junin, Guanarito, Machupo viruses and RNA-dependent RNA-polymerase for Lassa, Ebola and Marburg viruses) amino acid sequences analysis with subsequent transform of revealed unique peptides into due sets of oligonucleotides was used to design probes for hybridization and primers.

[On-microchip PCR for detection of influenza A viruses subtypes, circulating in the human population].

A oligonucleotide microchip was developed for revealing Influenza A viruses subtypes, circulating in human population: pandemic H1N1 swine influenza viruses, seasonal H1N1, H2N2, H3N2, H5N1, H9N2, H7N9. Typing of influenza virus was performed by on-microchip PCR. We used immobilized primers-probes selected for the neuraminidase gene that allows determining both subtype of neuraminidase and subtype of hemagglutinin.

Binding properties of the conjugates of oligo(2'-O-methylribonucleotides) with minor groove binders targeted to double stranded DNA.

Design, synthesis, physico-chemical and in vitro biological studies of new pyrimidine oligo(2'-O-methylribonucleotide) conjugates with oligocarboxamide minor groove binders (MGB) and benzoindoloquinoline intercalator (BIQ) are described. These conjugates formed stable triple helices with the target double-stranded DNA and inhibited its in vitro transcription upon binding.

Conjugates of oligo(2'-O-methylribonucleotides) with minor groove binders as new sequence-specific agents recognizing both grooves of double-stranded DNA.

Design, synthesis and physico-chemical studies of new pyrimidine oligo(2'-O-methylribonucleotide) conjugates with one or two oligo(pyrrolecarboxamide) minor groove binders (MGB) are described.

Stabilization of DNA double and triple helices by conjugation of minor groove binders to oligonucleotides.

New conjugates containing two parallel or antiparallel carboxamide minor groove binders (MGB) attached to the same terminal phosphate of one oligonucleotide strand were synthesized. The conjugates interact with their target DNA stronger than the individual components. Effect of conjugated MGB on DNA duplex and triplex stability and their sequence specificity was demonstrated on the short oligonucleotide duplexes and on the triplex formed by model 16-mer oligonucleotide with HIV polypurine tract.

Inhibition of human DNA topoisomerase I by new DNA minor groove ligands: derivatives of oligo-1,3-thiazolecarboxamides.

A series of novel thiazole-containing oligopeptides (oligo-1,3-thiazolecarboxamides) interesting specifically with the minor groove of DNA was shown to inhibit human DNA topoisomerase I (topo I). Inhibitory effects of thiazole-containing oligopeptides (TCO) increase with the number of thiazole units in such compounds. Inhibitory properties of TCO containing 3 or 4 thiazole units were shown to be 3-10 times better than those of the well-known natural antibiotic, distamycin A containing pyrrole rings. The structure of various additional groups attached to the N-terminus and C-terminus of TCO had no significant effect on TCO interaction with the complex of DNA and topo I. TCO were shown to be capable of binding with double-stranded DNA (dsDNA), and the majority of TCO analyzed were more effective in binding with dsDNA than distamycin A. Possible reasons for the different effects of distamycin A and TCO on the reaction of relaxation catalyzed by topo I are discussed.

Synthesis and evaluation of oligo-1,3-thiazolecarboxamide derivatives as HIV-1 reverse transcriptase inhibitors.

A set of oligo-1,3-thiazolecarboxamide derivatives able to interact with the minor groove of nucleic acids was synthesized. These oligopeptides contained different numbers of thiazole units presenting dimethylaminopropyl or EDTA moieties on the C-terminus, and aminohexanoyl or EDTA moieties on the N-terminus. The inhibition of such compounds on HIV-1 reverse transcriptase activity was evaluated using different model template primer duplexes: DNA x DNA, RNA x DNA, DNA x RNA and RNA x RNA. The biological properties of the thiazolecarboxamide derivatives were compared to those of distamycin, another minor groove binder which contains three pyrrole rings. Similar to distamycin, the thiazole containing oligopeptides were good inhibitors of the reverse transcription reaction in the presence of DNA x DNA. But in contrast to distamycin, the oligothiazolide derivatives were able to inhibit reverse transcription in the presence of RNA x DNA or DNA x RNA template primers. Both distamycin and oligothiazolecarboxamides had low affinity for RNA x RNA duplexes. The inhibition obtained with the newly synthesized thiazolecarboxamides showed that these compounds were more powerful and versatile inhibitors of the RT-dependent polymerization than the natural minor groove binder distamycin.

Inhibition of HIV-1 integrase-catalysed reaction by new DNA minor groove ligands: the oligo-1,3-thiazolecarboxamide derivatives.

Human immunodeficiency virus type 1 (HIV-1) integrase (IN) is an essential enzyme in the life cycle of the retrovirus, responsible for catalysing the insertion of the viral genome into the host cell chromosome. For this reason it provides an attractive target for antiviral drug design. We synthesized a series of novel thiazole (Tz)-containing oligopeptides (TCOs; oligo-1,3-thiazolecarboxamides), specifically interacting within the minor groove of DNA. The oligocarboxamide derivatives contained 1-4 Tz rings and different N- and C-terminal groups. The effect of these oligocarboxamides on the HIV-1 IN-catalysed reaction was investigated. Some of the compounds were able to inhibit the reaction. The inhibitory effect of the TCOs increased with the number of Tz units. The structure of various additional positively and/or negatively charged groups attached to the N- and C-termini of TCOs had a pronounced effect on their interaction with the DNA substrate complexed to IN. Modified TCOs having a better affinity for this complex should provide a rationale for the design of drugs targeting the integration step.

Specific features of electron distributions at altitudes of 400 km.

New experimental data obtained on the orbital station 'MIR' in 1991 during solar maximum are discussed. Electron fluxes with Ee>75 keV were registered for three different directions as well as for electrons with Ee>300 and 600 keV. Spatial and time distributions of electron fluxes in the trapping region are presented. In the inner radiation belt an additional maximum is observed at L=1.25-1.35, and the fluxes in the 22-05h MLT interval are 2-3 orders of magnitude smaller, than during other local times. In this region a flattening of the electron spectrum is observed. The results obtained were compared with the AE-8 model.

Comparative studies of gamma-interferon receptor-like proteins of variola major and variola minor viruses.

To study specific properties of the human gamma-interferon (gamma-IFN) receptor-like proteins of the highly virulent and low virulent strains of variola (smallpox) virus (VAR) recombinant plasmids determining synthesis of these proteins in E. coli cells have been constructed. The recombinant viral gamma-IFN receptor-like proteins have been found to have high interferon-neutralising activity with regard to human gamma-IFN but not murine gamma-IFN and human alpha-IFN. The variola major and variola minor proteins under study do not differ in the efficiency of human gamma-IFN antiviral activity inhibition.

Starting window, a distinct element in the cap-independent internal initiation of translation on picornaviral RNA.

Initiation of translation on picornaviral RNA templates occurs via cap-independent ribosome binding to a cis-acting element, internal ribosome entry site (IRES). Mapping of the starting point of translation relative to the IRES was attempted using Theiler's murine encephalomyelitis virus (TMEV) RNA as a model. The possibility that the starting point is determined by the conserved oligopyrimidine upstream of the initiator codon was studied. In contrast to poliovirus, neither the conserved oligopyrimidine nor an AUG at a fixed distance downstream of this oligopyrimidine are required for efficient translation of the TMEV RNA in Krebs-2 extracts or reticulocyte lysates or for viral infectivity; mutants lacking the oligopyrimidine/AUG tandem were stable upon passage in BHK-21 cells. A short template segment, the starting window, was defined, wherefrom ribosomes begin translation or downstream scanning depending, respectively, on the presence or absence of a good-context AUG within this window. Using a collection of the engineered TMEV mutant RNAs, the starting window was mapped to 16-17 nt downstream of the IRES and was found to be approximately a dozen nt long. The efficiency of translation initiation from an AUG linearly increased upon the 5'-->3' displacement of the initiator codon within the window. The competence of the starting window did not appear to depend markedly on its primary structure; however, it was completely inactivated ("closed") with concomitant dramatic inhibition of total protein synthesis upon conversion of the corresponding RNA segment into a double-stranded form.

Towards identification of cis-acting elements involved in the replication of enterovirus and rhinovirus RNAs: a proposal for the existence of tRNA-like terminal structures.

On the basis of a comparative analysis of published sequences, models for the secondary structure of the 3'-terminal [poly(A)-preceding] untranslated region of the entero- and rhinovirus RNAs were worked out. The models for all these viruses share a common core element, but there are an extra enterovirus-specific element and still an additional element characteristic of a subset of enterovirus RNAs. The two latter models were verified for poliovirus and coxsackievirus B genomes by testing with single-strand and double-strand specific enzymatic and chemical probes. A tRNA-like tertiary structure model for the 3'-terminal folding of enterovirus RNAs was proposed. A similar folding was proposed for the 3' termini of the negative RNA strands as well as for the 5' termini of the positive strand of all entero- and rhinovirus RNAs. Implications of these data for template recognition during negative and positive RNA strands synthesis and for the evolution of the picornavirus genomes are discussed.

Prokaryotic-like cis elements in the cap-independent internal initiation of translation on picornavirus RNA.

Initiation of translation on picornavirus RNAs is accomplished through internal binding of ribosomes to a complex cis-acting element. Here we show that efficient function of this element involves two appropriately spaced smaller elements: UUUCC and an AUG. This conclusion emerged from analysis of the genome structures of spontaneous revertants of mutant polioviruses with extended insertions between the UUUCC and AUG motifs. It was confirmed by the results obtained with specially designed constructs. A similarity to the prokaryotic translation initiation mechanism, which involves the Shine-Dalgarno sequence, is emphasized, but in the picornavirus system the position of the UUUCC must be strictly fixed relative to upstream cis-acting elements, and the AUG may not necessarily serve as an initiation codon.

Conserved structural domains in the 5'-untranslated region of picornaviral genomes: an analysis of the segment controlling translation and neurovirulence.

A model of secondary structure common for the central part (ca. 400 nucleotides) of the 5'-untranslated regions (5'-UTR) of all the so far sequenced genomes of polioviruses, coxsackieviruses, and rhinoviruses was derived on the basis of evolutionary and thermodynamic considerations. According to the model, this part of the genome comprises three domains, which appear to be involved, at least in the poliovirus genome, in the control of viral neurovirulence and in vitro translation. Some salient features of this model were supported by investigating RNAs of five poliovirus and one coxsackievirus strains with respect to their accessibility to modifications with dimethyl sulfate and sensitivity to single-strand- and double-strand-specific nucleases. In contrast to the previous suggestion, no major changes in the conformation of the Sabin vaccine poliovirus type 3 5'-UTR due to the transition in position 472 were observed. The biological relevance of the conserved primary and secondary structure elements in the picornaviral 5'-UTRs is discussed.