[Clinic-X-ray characteristics of periodontal disease in elderly patients.]

The significant prevalence of periodontal diseases in elderly patients makes the research relevant. By now, the issues of complex clinical and radiological semiotics of generalized periodontitis using high-tech research methods is not sufficiently studied. The research addressed the clinical picture and three-dimensional computed tomographic semiotics of severe chronic generalized periodontitis focusing 25 elderly patients with severe chronic generalized periodontitis. It verified the necessity to use an organ-oriented program of multiplanar (volumetric) cone-beam computed tomography coupled with the analysis of the research results, as well as a mandatory analysis of densitometry indicators of the jaw bone tissue in diagnostically significant periodontal zones.

Differences in the Differentiation Potential and Relative Levels of Gene Expression in the Bone Marrow-Derived Fibroblast Colony-Forming Units in Patients during the Onset of Aplastic Anemia Depending on the Disease Severity.

The differentiation potential of individual clones of fibroblast CFU (CFU-F) was studied and the relative expression level of genes was analyzed in the culture of CFU-F from the bone marrow in patients with non-severe and severe forms of aplastic anemia at the onset of the disease. The differentiation potential of CFU-F clones was determined by the relative expression of marker genes using quantitative PCR. In aplastic anemia, the ratio of CFU-F clones with different differentiation potential changes, but the molecular mechanisms of this phenomenon are different in non-severe and severe aplastic anemia. In the culture of CFU-F in non-severe and severe aplastic anemia, the relative expression level of genes associated with the maintenance of the hematopoietic stem cell in the bone marrow niche changes, but the decrease in the expression of immunoregulatory genes occurs in severe form only, which may reflect differences in the pathogenesis of non-severe and severe aplastic anemia.

[Genome Editing in Species of the Tribe Triticeae with the CRISPR/Cas System].

The tribe Triticeae includes important agricultural crops, such as bread wheat, durum wheat, barley, rye, and triticale. Research in the field of reverse genetics and genetic engineering of Triticeae received a new impetus as the CRISPR/Cas genome editing system came into broad use. The review describes and analyzes the data on recent advances in genomic editing of cultivated plants of the tribe Triticeae and tools used in the field. The tools most commonly used for genome editing in Triticeae include the codon-optimized Cas9 gene under the control of the maize ubiquitin gene promoter and guide RNAs under the control of Pol III promoters U6 and U3 in one or more binary vectors. Phosphinothricin and hygromycin resistance genes are used as selectable genes. Agrobacterium-mediated transformation and biolistics are performed to obtain genome-edited plants, and immature embryos are used as explants. Approaches developed to overcome the problem of low regenerative capacity of Triticeae include in planta transformation of shoot apical meristems, transformation of microspores and pollen grains, and the use of haploinductors. Bread wheat and barley were subject to genomic editing in the majority of studies published to date, and durum wheat and triticale were recently used in CRISPR/Cas knockout studies of target genes. Further progress in the development of genome editing of cultivated plants of the tribe Triticeae should be aimed at expanding the range of species and varieties involved and overcoming the problems of low regenerative capacity. This will allow genetic modification of elite varieties, which will be in demand in agricultural production.

Bone Marrow Multipotent Mesenchymal Stromal Cells from Patients with Aplastic Anemia Retain Their Ability to Support Hematopoietic Precursors despite Pronounced Changes in Gene Expression.

The properties of bone marrow-derived multipotent mesenchymal stromal cells (MSC) of patients with aplastic anemia at the onset of the disease are studied insufficiently. The aim of this work was to test the ability of MSC from patients with aplastic anemia to maintain hematopoietic precursors and to analyze the expression of genes associated with hematopoiesis and immune response. The ability of MSC to maintain hematopoietic precursors was determined by counting cobblestone area-forming cells; gene expression was analyzed by quantitative PCR. It was shown that MSC of patients with aplastic anemia preserve their ability to maintain hematopoietic precursors. Pronounced changes in the expression of the VEGFA and ANGPT1 genes were found. MSC from aplastic anemia patients with PNH clone significantly differ from those from aplastic anemia patients without PNH clone in terms of the expression of the SDF1, IL1R, and VEGFA genes. Changes in gene expression can be associated with the pathogenesis of the disease.

A Model of Short Bowel Syndrome in Rodents in a Long-Term Experiment.

Simulation of short bowel syndrome (SBS) in experimental animals is of great interest for studies in translational medicine. The processes of intestinal adaptation are most often studied in rats aged 8-10 weeks with up to 70% resection the small bowel. In this case, the animals are euthanized in 2 weeks after the surgery; further development of the pathological process remains practically unexplored. In our experiment, a number of blood biochemical parameters and morphological signs of intestinal adaptation were assessed within 34 weeks after surgery. It was shown that after resection of 70% of the small bowel in rats (180±15 g), clinical symptoms are observed only during the first 2 weeks, however, the mortality in the group was observed throughout the entire period of the study in the absence of clear clinical signs. Morphological compensatory changes in the walls of the small intestine continued up to 34 weeks after resection, and their morphometry can be used to describe adaptation in the dynamics of the study.

[Implementation of the prevention program in the school dentistry system in the context of healthcare modernization]. / Realizatsiya programmy profilaktiki v sisteme shkol'noi stomatologii v usloviyakh modernizatsii zdravookhraneniya.

THE AIM OF THE STUDY: The aim of the study was to develop and implement a program for the prevention of dental diseases for school-age children based on an individual approach to treatment and preventive measures. MATERIAL AND METHODS: 1848 children aged from 6 to 17 were examined. The main observation group consisted of children from school No. 1694 in the South-Western Administrative District of Moscow (935 children), the comparison group with no prevention program consisted of children from school No 1206 (913 children) of the same Moscow district. The effectiveness of the prevention program was assessed in key age groups of children 6, 12 and 15 years old. The prevention program included the study of dental morbidity, assessment of the level of oral hygiene of children, parents and teachers, conducting sanitary and educational work, teaching children hygiene and monitoring its implementation, developing and implementing the individual plan of therapeutic and preventive measures for children of all age groups. RESULTS: As a result of the prevention program, the number of healthy children without caries increased. The introduction of a prevention program showed its effectiveness, the reduction of caries growth among 12-years-old children who participated in the prevention program over 5 years was 37%, and the number of healthy children increased by 15%. CONCLUSION: Carrying out a prevention program at school has shown that coordinated actions of parents, teachers and dentists lead to an improvement in the condition of hard tissues of the permanent teeth of schoolchildren, a decrease in the risk of inflammatory periodontal diseases in adolescence, and an improvement in the quality of individual oral hygiene. Of particular importance is the presence of a dental office and a dental hygienist in the school, for the possibility of carrying out preventive measures for children of all age groups.

Intracortical Directed Connectivity for Information Retention in Visual-Spatial Working Memory.

A group of 27 healthy young adults solved a task involving the working memory (WM) activation, consisting of the comparison between the spatial orientations of two sequentially presented square-wave luminance gratings. We investigated the effective (directed) connectivity patterns between the frontal and postcentral cortical regions related to the visual system. The connectivity was assessed using vector autoregression modeling of EEG. It was shown that the strength of the top-down right-hemispheric connectivity patterns directed from the frontal cortex to the visual areas in θ frequency was significantly lower at the stage of stimulus retention in the WM than at the stage of stimulus anticipation. On the contrary, in the α band the descending influences were slightly more intense. The results of the study showed the frequency-dependent dynamics of the descending influences of the frontal cortex on visual areas and confirm that the frontal cortex plays the role of a controlling and modulating center in the brain system underlying WM.

[A case report of familial dyskeratosis congenital. Case report].

Dyskeratosis congenita (DC) is a hereditary syndrome of bone marrow failure, which develops because of telomeres defects and combines with cancer predisposition. Its classical clinical features are skin pigmentation, nail dystrophy, oral leukoplakia (skin-mucosa triad). The goal is to describe the algorithm of diagnosis, clinical specificities of DC and specific treatment for cases of DC in one family. The present report includes descriptions of diagnosis and treatment of family members diagnosed for the first time as having a DC. The report shows an importance of all diagnostic stages: from a medical history and clinical picture to an application of modern high-tech diagnostic methods (flow-FISH, NGS). The report underlines an importance of diagnosis of all family members for excluding an asymptomatic form after a case of DC has been already detected in that family. A high frequency of a toxicity and secondary neoplasia makes it necessary to realize an individual approach at treatment of each patient with DC (the earliest start of androgen treatment, prompt decision of implementation of allogenic hematopoietic stem cell transplantation). The knowledge of pathogenesis, clinical features and principles of diagnosis and therapy of this disease is relevant to pediatricians and hematologists.

Local immunity and microflora of the reproductive tract of gynecological patients before and after immunomodulatory therapy.

Immunomodulatory drugs are widely used as drugs for the treatment of inflammatory diseases of microbial etiology in clinical medicine. The authors evaluated the effectiveness of the therapeutic effect of the Cycloferon immunomodulator in the treatment of chronic purulent-inflammatory diseases of the reproductive tract using clinical, microbiological and immunological parameters. It was shown that under the influence of immunomodulatory therapy, the restoration of the qualitative and quantitative composition of the normoflora was observed with the elimination of etiologically significant microorganisms. The immunocorrective effect of therapy on the indices of local immunity was established: the concentration of lactoferrin increased in the cervical secretion, the level of cytokines IL-1ß and γ-IFN normalized, the amount of secretory IgA increased significantly, which contributed to the enhancement of local protective reactions, as well as the clinical efficacy of therapy, which was manifested in the reduction and/or disappearance of pain syndrome and the absence of relapse for 2 or more years. Conducted researches allow us to speak of immunotropic drugs as a promising direction in the treatment of inflammatory processes of the reproductive tract, a significant advantage of which is rational immunomodulation directly in the focus of inflammation, the availability of treatment, the absence of side effects with adequate therapeutic regimens, which makes it relevant to further study this direction of immunocorrection and its implementation into wide clinical practice.

Comparison of polymerase chain reaction and flow cytometry for measuring telomere length of human leukocytes.

Telomere length can be measured by polymerase chain reaction (PCR), allowing to obtain the absolute length of telomeres (ALT) in base pair, and by flow cytometry, which can only estimate the relative telomere length. The aim of the study was to compare the results of the two methods and to develop an accurate and reliable way of converting the relative telomere length to absolute. The peripheral blood from 21 donors was analyzed. Measurement of leukocyte telomere length by flow cytometry was carried out using a commercial Telomere PNA Kit / FITC (Dako, Denmark) with two CytoFLEX flow cytometers (Beckman Coulter, China) and BD FACSCanto II (Becton Dickinson, USA), obtaining the molecular equivalent of fluorescence (MEF). To measure telomere length by real-time PCR, calibrators with a known number of telomeric repeats were prepared. Two quantitative PCRs were carried out: one for telomeric repeats, the other for determining the number of genome-equivalents of DNA, three times for each sample, which made it possible to calculate ALT. A strong direct relationship was found between the MEF obtained with BD FACSCanto II and CytoFLEX (r = 0.97). Analysis of PCR and flow cytometry results showed a significant correlation between ALT and MEF. We calculated the regression equations of ALT and MEF for CytoFLEX - y = 0.0043x (r = 0.84) and for BD FACSCanto II - y = 0.0051x (r = 0.82). Correlation analysis showed a high comparability of telomere lengths measured by two methods. The obtained regression equations allow converting the results of flow cytometry into absolute values, allowing the comparison of the results of different research groups and the use of this method in clinical trials.

Alpha-B-Crystallin Effect on Mature Amyloid Fibrils: Different Degradation Mechanisms and Changes in Cytotoxicity.

Given the ability of molecular chaperones and chaperone-like proteins to inhibit the formation of pathological amyloid fibrils, the chaperone-based therapy of amyloidosis has recently been proposed. However, since these diseases are often diagnosed at the stages when a large amount of amyloids is already accumulated in the patient's body, in this work we pay attention to the undeservedly poorly studied problem of chaperone and chaperone-like proteins' effect on mature amyloid fibrils. We showed that a heat shock protein alpha-B-crystallin, which is capable of inhibiting fibrillogenesis and is found in large quantities as a part of amyloid plaques, can induce degradation of mature amyloids by two different mechanisms. Under physiological conditions, alpha-B-crystallin induces fluffing and unweaving of amyloid fibrils, which leads to a partial decrease in their structural ordering without lowering their stability and can increase their cytotoxicity. We found a higher correlation between the rate and effectiveness of amyloids degradation with the size of fibrils clusters rather than with amino acid sequence of amyloidogenic protein. Some external effects (such as an increase in medium acidity) can lead to a change in the mechanism of fibrils degradation induced by alpha-B-crystallin: amyloid fibers are fragmented without changing their secondary structure and properties. According to recent data, fibrils cutting can lead to the generation of seeds for new bona fide amyloid fibrils and accelerate the accumulation of amyloids, as well as enhance the ability of fibrils to disrupt membranes and to reduce cell viability. Our results emphasize the need to test the chaperone effect not only on fibrillogenesis, but also on the mature amyloid fibrils, including stress conditions, in order to avoid undesirable disease progression during chaperone-based therapy.

[Design of Guide RNA for CRISPR/Cas Plant Genome Editing].

CRISPR/Cas technology of genome editing is a powerful tool for making targeted changes in the DNA of various organisms, including plants. The choice of the precise nucleotide sequence (protospacer) in the gene to be edited is important in the design of guide RNA, which can be carried out by specialized software. We review and compare all the known on-line and off-line resources for guide RNA design, with special attention paid to tools capable of searching for off-target edits sites in plant genomes. The use of Cas12a may be preferable to Cas9. Techniques allowing C→T and G→A base editing without DNA cleavage are discussed along with the basic requirements for the design of effective and highly specific guide RNAs. Ways for improving guide RNA design software are presented. We also discuss the lesser risks of off-target editing in plant genomes as opposed to animal genomes. Examples of edited plant genomes including those that do not lead to the creation of transgenic plants are reviewed.

[Endothelial damage and circadian blood pressure profile in rheumatoid arthritis].

AIM: To study the influence of the state of endothelium on the daily profile of arterial pressure (AP) in patients with rheumatoid arthritis (RA). MATERIALS AND METHODS: In 70 RA pts carried out C-reactive protein (CRP), vascular endothelial adhesion molecule type 1 (sVCAM-1), antigen von Willebrand Factor (AG WF), interleukin-8 (Il-8), rheumatoid factor (RF), IgG, endotheline-1 (ET-1), number of desquamated endotheliocytes cells (DE), VS, activity of renin by immunoenzyme analysis. The dysfunction of endothelium was evaluated by calculation of DE. The functional methods included the daily monitoring of arterial pressure (AP). RESULTS: Arterial hypertension (AH) occurred in 40 (57.1%) pts. RA pts are revealed the signs of endothelial dysfunction, about which significant differences among the indices of activation of endothelium in comparison with control group testify. ET-1, sVCAM-1, vWF AG, Il-8, CRP content was higher in RA pts. Reliably above there was a number of DE. Reliable differences according to these indices depending of RA activity were discovered. With conducting of correlation analysis it is revealed, that markers of the activation of endothelium: sVCAM-1, vWF AG positively correlated with increasing RF IgG and indices of the immune inflammation: CRP, and DE number. In patients suffering from RA, showed signs of endothelial dysfunction. The positive correlation between endothelial damage and daily profile of AP show the relationship of these processes. CONCLUSION: Positive correlations between the damage of endothelium and disturbance of AP daily profile testify about the interrelation of these processes.

Working Memory for Low-Level Visual Features: Sensory Mechanisms for Detecting the Mismatch between the Current Orientations and Those Stored in Memory.

The amplitudes of the P100 and N150 early components of evoked potentials in the visual cortex have been analyzed on 33 volunteers with normal vision during matching between the current orientation and that stored in memory. An increase in the P100 response in the occipital and parietal cortical areas was identified as an informative indicator of mismatch between the current and stored-in-memory orientations. This effect was not found for more complex stimuli, namely, spatial patterns. The N150 component demonstrated a similar effect, but in contrast to P100 it was not stimulus specific. Thus, in the first 100 ms, a signal of mismatch between the current and stored-in-memory orientations arises in the early visual areas that represents a mechanism for early implicit response to changes in the basic characteristics of the visual space.

[The capacity of Escherichia coli to form biofilms in presence of nano-particles of aluminum oxide.]

The effect of various concentrations of suspensions of nanoparticles of aluminum oxide of size of 30 and 70 nm on capacity of Escherichia сoli to form biofilm in vitro was examined. The study used highly virulent strain of Escherichia сoli isolated from a patient with chronic pyelonephritis. The strain was characterized by high capacity of forming biofilm. It is established that nanoparticles of aluminum oxide with size of 30 nm inhibited capacity of forming biofilm. At that, a reliable decreasing of analyzed indication in two times occurred and analyzed clones of Escherichia сoli passed to category of microorganisms with average capacity of forming biofilm. It is established that intensity of factor suppression depended both on size and concentration of nanoparticles in medium. The most effective in suppression of capacity of forming biofilm was concentration of nanoparticles of aluminum oxide 0,015 mkg/ml that decreased intensity of factor in two times.

[The characteristics of microbiota of periodontal recesses in smoking patients with chronic generalized periodontitis].

The clinical examination of 36 tobacco smokers with chronic generalized periodontitis of light, average and severe degree was carried out. The examination established poor hygienic condition of oral cavity, less expressed inflammatory reaction of tissues of periodont and predominance of occurrences of destruction of alveolar portion of bone as compared with the group of 59 non-smoking patients with chronic generalized periodontitis of light, average and severe degree. The study demonstrated higher rate of detection of T. forsythia in smokers as compared with non-smoking patients at all stages of of development of chronic generalized periodontitis. Under light stage of chronic generalized periodontitis increasing of rate of detection of T. forsythia more than twice was registered. P.gigngivalis and P.intermedia were detected in smoking patients with light stage of chronic generalized periodontitis either in the same values or more rarely as compared with non-smokers. In the group of smokers with average stage of chronic generalized periodontitis increasing of rate of occurrence of association of T. forsythia-P. gigngivalis-P. intermedia occurred more than five times in comparison with non-smokers. The obtained results indicate on relationship between alterations of microbiota and aggressive development of chronic generalized periodontitis in smoking patients and on development in periodontal recesses of smokers of favorable conditions for growth of T. forsythia. The presence of T. forsythia is a significant factor of development of destructive processes in tissues of periodont.

[Variations of oral fluid microbiota in healthy children and adolescents]. / Varianty mikroflory rotovoi zhidkosti u prakticheski zdorovykh detei i podrostkov.

The aim of the study was to establish the bacterial and viral associations of saliva and the immune resistance in 127 healthy people of different ages. The analysis sorted out three variants of oral fluid microbiota differing by certain combination of indigenous and facultative microflora. It was found that with age there is a significant increase in the number of adolescents with the third variant of the microbiota, characterized by a decrease in the number of indigenous microflora and increase in opportunistic microorganisms and viruses. With an increase in microecological shifts a tendency to decrease the level of lysozyme, an increase in the number of secretory IgA, accompanied by a decrease in antibody was revealed. It has been shown that bacterial and viral component of the microbiota of the oral fluid are in a dynamic relationship with each other, as well as immune resistance and can serve as an indicator of the health level and the selection criteria at the stages of clinical examination of children and adolescents, with the risk of dental disease.

CHARACTERISTICS OF AMYLOID DEPOSITS DETECTED IN THE INTERNAL ORGANS OF mdx MICE.

The dystrophin-deficient mdx mouse is the most commonly used experimental model of Duchenne muscular dystrophy (DMD). Although the amyloid has been shown in the muscle biopsies of patients with different types of muscular dystrophies, there are no data on the amyloid accumulations in the biopsy of DMD patients or mdx mouse. Therefore, the aim of the present study was to testify the hypothesis of probable accumulation of amyloid in the visceral organs of mdx mouse. Specimens of myocardium, kidneys, and liver of male and female mdx mice aged from 2 months to 1.5 years (n = 9) were used in the study. The histochemical staining with Congo red demonstrated amyloid accumulations in the studied organs of the mdx mice. Morphology and localization of the found accumulations were described in details and analyzed. The mass-spectrometric study determined the vitronectin and apolipoprotein A-II as the most probable components of the amyloid accumulations in the mdx mouse.

Cytokine-producing function of human blood cells in chronic atrophic gastritis and gastric adenomas and adenocarcinomas.

The cytokine-producing potential of blood cells has been studied in the atrophic gastritis-adenoma-adenocarcinoma progression of pathological states of the stomach. It has been revealed that, at the initial stage of carcinogenesis, namely adenoma, immunocompetent cells have the highest cytokine-producing proto-oncogenic potential as compared to both atrophic gastritis, which presents a precancerous condition, and completely formed malignant tumor (gastric adenocarcinoma).

Orosensory Food Testing in Fish: Chronology of Behavior.

The orosensory food testing behavior in fish was studied using the nine-spined stickleback Pungitiuspungitius as an example. The patterns of the chronology of manipulations performed by fish in testing food objects were identified. The existence of two stereotypical patterns of feeding behavior was confirmed, and their new characteristics were obtained. The relationship between the responsiveness to food and the predisposition of fish to feeding was revealed.