Половые отличия пула свободных аминокислот-нейромедиаторов у крыс Крушинского-Молодкиной

The study of the role of neurotransmitter systems in the pathogenesis of epilepsy is one of the priorities of epileptology. New data on the functions of free neurotransmitter-like amino acid in the central nervous system are of the greatest importance and determine the prospects for the development of novel effective anticonvulsants. It is widely believed in clinical medicine that epilepsy has distinct gender characteristics. The aim of this study was to investigate the gender peculiarities in the content of neurotransmitter amino acids in the brain of Krushinsky-Molodkina (KM) rats, which were used as model organisms for the study of genetically induced audiogenic epilepsy. The content of Asp, Glu, GABA, Gly, and Tau of the medulla oblongata, hippocampus and cerebral cortex were determined using high-performance liquid chromatography (HPLC) in intact KM rats, KM rats exposed to a series of epileptiform seizures, and Wistar rats (control group). Both the Wistar and KM rats had gender distinctions in the distribution of free amino acids among the investigated brain parts. The audiogenic epilepsy was characterized by smoothing gender differences as well as differences between the concentrations of free amino acids in the cortex and medulla oblongata, specific for Wistar rats. The changes observed in male rats after the set of seizures included the increase in GABA concentration and a decrease in the Gly level in all investigated brain parts, as well as the decrease of the Tau content in the cortex and hippocampus. At the same time, the Glu content in cortex increased, while the Asp level decreased. After 6 days of audiogenic stimulations the female KM rats demonstrated the increase in the Glu level in all investigated brain parts, the increase in Gly and Asp levels in hippocampus, and no changes in the GABA content. Thus, after the set of epileptiform seizures the KM rats achieved a new steady state of the studied amino acids pool, which differed in males and females. In this case, gender differences significantly changed after the seizures. © 2020 Russian Academy of Medical Sciences. All rights reserved.The work was performed within the framework of the state task of the IIF UB RAS (Registration number AAAA-A18-118020590108-7)

Achievements and prospects of applying high-throughput sequencing techniques to potato genetics and breeding

In recent years, marker-assisted selection (MAS) has been intensively used to increase potato breeding efficiency. Large-scale studies of the potato genome and genes exploiting next-generation sequence (NGS) approaches are required for broad application of MAS, genomic selection as well as genomic editing (the newest approach for creating potato with desired properties). In this review, trends in potato NGS-based research are overviewed, and related Internet resources are systematized. Special attention is given to peculiarities of the models and the approaches used in potato genetic studies, taking into account the complex organization of its genome and a high level of heterozygosity. In genetic studies diploids are used often, including diploid potato species, artificially obtained heterozygous dihaploids and homozygous double monoploids. The availability of artificially created diploid forms played an essential role in potato genome sequencing, which was completed in 2011. The Potato Genome Sequencing Consortium activities included not only constructing genome libraries, sequencing, assembling and annotation of the genome, but also genome sequence-based investigations uncovering features of potato genome evolution, SNP identification, analysis of genes and gene networks regulating resistance to phytopathogens and technological characteristics. An important outcome of the genome sequencing was further identification of more than 8 thousand SNPs and approbation of the Genotyping-by-sequencing (GBS) method on potato, which is the basis for potato genomic selection and for discovery of economically important genes using genome wide association studies (GWAS). Optimization of existing bioinformatic tools to support these studies, taking into account potato genome organization peculiarities, are carried out. This review gives analysis of databases containing potato genome and transcriptome sequencing results, as well as accompanying resources. This information should prove useful while planning comparative assays of potato transcriptome or application of DNA-markers. Sequencing of the genome as well as transcriptomes and microRNomes of cultivated potato and its wild relatives, on one hand, is of fundamental interest, assisting in detecting features of genome evolution, ontogenetic development and mechanisms of various environmental stresses responses. On the other hand, it is the basis for a wide range of practical applications for developing effective genomic and gene-specific markers and marker-assisted breeding of new potato cultivars with desired properties

Efficacy Evaluation of Combination Therapy in Patients with Generalized Periodontitis According to the Index Estimation of Periodontal Status: Experimental Randomized Single-Arm Study

Background. Treatment and prevention of inflammatory periodontal diseases represent a serious general medical and social problem, which remains relevant due to the significant prevalence of periodontitis among the population, as well as the associated loss of teeth and the negative impact of periodontal foci of infection on the body as a whole. Chronic generalized periodontitis is difficult to treat: it is only possible to achieve periods of remission. Therefore, it is of great theoretical and practical importance to develop further ways to improve methods for treating inflammatory-dystrophic damage of periodontal tissues in chronic generalized periodontitis.Objective. To perform an index estimation of treatment efficacy in patients with moderately severe chronic generalized periodontitis.Methods. An experimental randomized single-arm study was conducted examining 204 patients diagnosed with moderately severe chronic generalized periodontitis. Patient examination and data analysis were carried out at the Dentistry Department of the Stavropol State Medical University and the Department for General Practice Dentistry of the Kuban State Medical University in 2021–2022. The study included patients aged 35 to 65 years diagnosed with moderately severe chronic generalized periodontitis. Patients with somatic diseases in the decompensation stage were not eligible for the study. The patients were randomly divided into three groups (closed-envelope method). Group 1 was treated using conventional methods. As well as conventional therapy, Group 2 was prescribed vitamin D (Aquadetrim® at a dose of 5000 IU once per day while monitoring vitamin D blood level). The third group of patients was prescribed conventional therapy along with the developed pharmacotherapy, which included vitamin D (Aquadetrim® at a dose of 5000 IU once per day), vitamin A (3.44% oil solution of retinol acetate at a dose of 50 000 IU, 15 min after a morning meal), and 2-ethyl-6-methyl-3-hydroxypyridine (Mexidol® at a dose of 125 mg twice a day). In each patient, the treatment efficacy was analyzed in adjacent oral cavity segments (split-mouth design) according to clinical pocket depth, attachment loss, and bleeding on probing. Also, the authors used the X-ray index to assess the condition of osseous structures and the simplified oral hygiene index. The specified indices were estimated prior to treatment and at one and 12 months following treatment. The statistical analysis of study results was performed using Excel Microsoft Office 2016 (Microsoft, USA).Results. The obtained data indicate that the immediate clinical outcomes (after one month) of combination therapy in patients from all three groups were approximately the same. However, the efficacy of treatment measures in the long-term period (12 months) was different, as evidenced by the dynamics of index values. Only in Group 3 patients, the full implementation of pharmacotherapeutic measures as part of the combination therapy of periodontitis ensured a stable therapeutic effect throughout the entire period of observation.Conclusion. The data obtained 12 months following treatment indicate that the additional prescription of the developed pharmacotherapy contributed to prompt inflammation relief in periodontal tissues, providing a stable treatment outcome and long-term remission

Index Evaluation of Clinical Efficacy of Immunomodulatory and Osteotropic Therapy for Chronic Generalized Periodontitis of Varying Severity: A Prospective Cohort Study

Background. The significant prevalence of chronic generalized periodontitis, the severity of its course, the increase in the number of complicated forms and short-term remissions cause a constant search for new methods and means of its treatment. When choosing the most effective methods of therapy, specialists mostly prefer a complex effect on the main pathogenetic links of the disease, while often losing sight of the body's own defenses, especially in terms of pharmacotherapeutic support and immunity strengthening. This study presents the results of a study of the clinical efficacy of the treatment of chronic generalized periodontitis by means of the developed therapy complex, including the use of immunomodulatory and osteotropic drugs.Objective — to study clinical efficacy of the developed complex immunomodulatory and osteotropic treatment for chronic generalized periodontitis.Methods. A prospective cohort study was conducted in 154 patients aged 20 to 75 with periodontitis of varying severity. Conventional examination of patients was carried out in specialized dental clinics DentoProma (Krasnodar) and Dentalife (Stavropol) based at the Dentistry Department, Stavropol State Medical University (Russia). The study period was 24 months. The patients were divided into a main group of 118 participants and a control group of 36 participants. The comprehensive treatment plan, prescribed to all patients, included proper oral hygiene education, individual and professional dental hygiene, topical anti-inflammatory agents, Vector therapy, open flap debridement, flap surgery, relevant splinting prosthetics (if necessary). The authors developed a comprehensive therapy, which included administration of the immunomodulatory agent Hepon (Immapharma Company, Russia), glucosamine hydrochloride and chondroitin sulfate sodium. All patients of the main and control groups were additionally ranked into subgroups depending on the severity of diagnosed periodontitis: slight (subgroup 1), moderate (subgroup 2) and severe (subgroup 3). The major focus of the study was to establish efficacy of the developed therapy which was defined by the degree of resolution of the inflammatory process and increase in the period of remission. Reflecting the presence and degree of inflammation, dental plaque index and oral hygiene status were considered to be the target indicators. Analysis and statistical processing of the obtained data were carried out using Microsoft Excel (Microsoft, USA).Results. The analysis of the data revealed that the immediate clinical results of the developed complex therapy both in the main group and in the control group were approximately identical. According to the follow-up data obtained one month after the treatment, additional prescription of complex osteotropic treatment to the patients with chronic generalized periodontitis contributed to rapid resolution of infection in the gingival tissues and provided stable treatment results.Conclusion. Introduction of the complex clinical and hygienic treatment promoted resolution of the inflammatory process in the gingival tissues of all patients who received immunomodulatory and osteotropic therapy

Содержание фактора активации тромбоцитов в плазме крови больных бронхиальной астмой. Влияние тромбоцитафереза

Levels of serum PA F were investigated in patients with various bronchial asthma (BA) forms and in various forms of the clinical manifestation of the disease before and after plateletapheresis (PtA). 15 patients with BA (9 atopic and 6 aspirin-sensitive ones) and 4 healthy donors were examined. The examination demonstrated that only tracks of PA F in the healthy donors are registered but in BA patients the PAF levels were significantly increased. It was found, that in patients with atopic BA the PAF levels were almost by two times higher than in aspirinic BA ones. Moreover, the PAF level in a great extent depends on the disease degree. The serum PAF level was decreased in 42% in average after trobmocytapheresis.The study demonstrates the features of thrombocytapheresis action and develops the imagination about its action mechanisms.Исследовался уровень фактора активации тромбоцитов (ФАТ) в крови больных различными формами бронхиальной астмы разной степени тяжести и влияние тромбоцитафереза на этот показатель. Обследовано 15 больных с БА (9 с атопической и 6 с аспириновой) и 4 здоровых донора. Исследования показали, что в крови здоровых доноров регистрируются лишь следы ФАТ, тогда как у больных БА содержание ФАТ значительно повышено. Выявлено, что у больных аспириновой астмой уровень ФАТ почти в 2 раза ниже, чем в группе больных атопической астмой. Кроме того, чем тяж елее протекает заболевание, тем вы ш е уровень ФАТ. После тромбоцитафереза уровень ФАТ в крови больных, получавших данную терапию , снизился в среднем на 42%.Исследования раскрывают особенности действия тромбоцитафереза, углубляют представления о механизме действия этой процедуры

Risk factors for cardiovascular diseases in patients received complex treatment for cranial and craniospinal tumors in childhood

Aim. To study the traditional risk factors for cardiovascular disease (CVD), indicators of endothelial function and exercise tolerance in patients received complex treatment for cranial and craniospinal tumors in childhood, including radiation therapy.Material and methods. We compared examination data of 48 patients who underwent treatment for brain tumors using cranial and craniospinal irradiation in childhood (mean age, 21,7±4,3 years, mean period after the end of treatment, 6,9±5,4 years), and 20 healthy volunteers. Examination methods included assessment of lipid profile, vascular stiffness and endothelial function using the Photoplethysmography and occlusion test, cardiopulmonary test, and in patients who underwent craniospinal irradiation, also echocardiography and duplex ultrasound of extracranial arteries.Results. Compared to healthy individuals, patients after a brain tumor were found to have lower blood pressure, higher heart rate (HR), significantly lower exercise performance (peak oxygen consumption, 19,8±6,4 ml×min-1×kg vs 30,3±5,8 ml×min-1×kg, p<0,0001) and a higher prevalence of dyslipidemia (56% vs 5%, p<0,0001), as well as an increase in the augmentation index, indicating higher stiffness of large vessels (-7,3±16,3 vs -20,3±7,9, p=0,001), and a trend towards a decrease in the occlusion index (p=0,051). Echocardiography and duplex ultrasound revealed no radiation-associated abnormalities.Conclusion. Determining the mechanisms and prognostic significance of the identified risk factors for CVD (dyslipidemia, decreased exercise tolerance, increased heart rate and vascular stiffness) in this category of patients requires further research. Regular monitoring of risk factors, primarily the lipid profile, and the use of preventive measures for individuals with an increased risk of CVD should be recommended

Novel experimental setup for megahertz X-ray diffraction in a diamond anvil cell at the High Energy Density (HED) instrument of the European X-ray Free-Electron Laser (EuXFEL)

The high-precision X-ray diffraction setup for work with diamond anvil cells (DACs) in interaction chamber 2 (IC2) of the High Energy Density instrument of the European X-ray Free-Electron Laser is described. This includes beamline optics, sample positioning and detector systems located in the multipurpose vacuum chamber. Concepts for pump-probe X-ray diffraction experiments in the DAC are described and their implementation demonstrated during the First User Community Assisted Commissioning experiment. X-ray heating and diffraction of Bi under pressure, obtained using 20 fs X-ray pulses at 17.8 keV and 2.2 MHz repetition, is illustrated through splitting of diffraction peaks, and interpreted employing finite element modeling of the sample chamber in the DAC

Insight into the Regulation of Glycan Synthesis in Drosophila Chaoptin Based on Mass Spectrometry

BACKGROUND: A variety of N-glycans attached to protein are known to involve in many important biological functions. Endoplasmic reticulum (ER) and Golgi localized enzymes are responsible to this template-independent glycan synthesis resulting glycoforms at each asparagine residues. The regulation mechanism such glycan synthesis remains largely unknown. METHODOLOGY/PRINCIPAL FINDINGS: In order to investigate the relationship between glycan structure and protein conformation, we analyzed a glycoprotein of Drosophila melanogaster, chaoptin (Chp), which is localized in photoreceptor cells and is bound to the cell membrane via a glycosylphosphatidylinositol anchor. Detailed analysis based on mass spectrometry revealed the presence of 13 N-glycosylation sites and the composition of the glycoform at each site. The synthetic pathway of glycans was speculated from the observed glycan structures and the composition at each N-glycosylation site, where the presence of novel routes were suggested. The distribution of glycoforms on a Chp polypeptide suggested that various processing enzymes act on the exterior of Chp in the Golgi apparatus, although virtually no enzyme can gain access to the interior of the horseshoe-shaped scaffold, hence explaining the presence of longer glycans within the interior. Furthermore, analysis of Chp from a mutant (RNAi against dolichyl-phosphate alpha-d-mannosyltransferase), which affects N-glycan synthesis in the ER, revealed that truncated glycan structures were processed. As a result, the distribution of glycoforms was affected for the high-mannose-type glycans only, whereas other types of glycans remained similar to those observed in the control and wild-type. CONCLUSIONS/SIGNIFICANCE: These results indicate that glycan processing depends largely on the backbone structure of the parent polypeptide. The information we obtained can be applied to other members of the LRR family of proteins