Outcome of the entomological monitoring for Crimean-Congo haemorrhagic fever virus in the western and southern regions of Kazakhstan in 2021-2022.

The natural foci of Crimean-Congo haemorrhagic fever (CCHF) in Kazakhstan are geographically located in the southern regions of the country (Kyzylorda, Turkestan and Zhambyl regions), where the infection of ticks with the CCHF virus predominantly reside, tick species composition and the number of vectors are monitored annually. The objective of our research was to investigate the genetic variants of the CCHF virus in the southern endemic regions, as well as to monitor the spread of the CCHF virus in the western regions of the country (Aktobe, Atyrau and Mangystau regions). In total, 974 (216 pools) ticks from the western regions and 3527 (583 pools) ticks from the southern regions collected during 2021-2022 were investigated. The presence of CCHF virus was detected by real-time reverse transcription PCR (qRT- PCR) in 1 pool out of 799 pools (0.12%) with Hyalomma scupense ticks captured in the CCHF-endemic Kyzylorda region. In the western regions, CCHF virus was not detected in ticks. The sequencing of incomplete fragments of the S, M and L segments of the CCHF virus in the detected virus was identified as genotype Asia - I. Phylogenetic analysis showed that the isolate obtained in this study is grouped with the isolate from a patient with CCHF, which we reported in 2015 (KX129738 Genbank). Our findings highlight the importance of including sequencing in the annual monitoring system for better understanding the evolution of the CCHF virus in the study areas of our country.

Characterization of tularemia foci in the Republic of Kazakhstan from 2000 to 2020.

The wide distribution of tularemia in the territory of Kazakhstan is associated with landscape and geographical characteristics. This is explained by a combination of natural factors: the presence of certain types of rodents-reservoirs and sources, ectoparasites-carriers of the causative agent of tularemia. The study of the current spatial and temporal characterization of tularemia in Kazakhstan from 2000 to 2020 will determine the epidemiological status of tularemia and improve the monitoring system in Kazakhstan. In this work we demonstrated the results of a retrospective survey of natural foci of tularemia: analysis of vector, small mammal and human data. The spatial and temporal characteristics of tularemia from 2000 to 2020 in the territory of Kazakhstan were studied in comparison with historical data, including the description of tularemia outbreaks, the clinical picture, and the source of infection, transmission factors, and geographical coordinates of outbreak registration. Sampling was carried out by trapping rodents on snap traps and collecting ticks by rodent combing and by "flagging" methods. For the last 20 years, 85 human cases of tularemia have been reported. During the period from 2000 to 2020, more than 600 strains of F. tularensis were isolated from field rodents and ticks in the natural foci of tularemia. MLVA typing of F. tularensis strains isolated from natural foci of tularemia in Kazakhstan over the past 20 years. The results of retrospective monitoring indicate that currently active foci of tularemia include the Aktobe, West Kazakhstan, Almaty, East Kazakhstan, and Pavlodar regions. Low-activity natural foci are located in the territory of the Akmola, Karaganda, North Kazakhstan, Kostanay, Atyrau, Zhambyl, and Kyzylorda regions. There are no active natural foci of tularemia in the Mangystau and Turkestan regions. The widespread occurrence of tularemia in the country is associated with landscape and geographical features that contribute to the circulation of the pathogen in the natural focus. An analysis of natural foci of tularemia showed that it is necessary to continue monitoring studies of carriers and vectors for the presence of the causative agent of the F. tularensis, in order to prevent mass cases of human disease.

GIS TECHNOLOGIES IN THE STUDY OF NATURAL RESULTS ESPECIALLY DANGEROUS DISEASES IN KAZAKHSTAN.

This study explores the application of GIS technologies in analyzing and visualizing spatial structures of especially dangerous infections (EPI) in Kazakhstan. International collaborations have facilitated projects studying the focal patterns of diseases, improving data analysis and visualization. Extensive electronic databases resulting from field research on EPI foci have elevated the study's depth. The dynamics of natural foci, influenced by intraspecific structures of infection carriers, are impacted by industrial and agricultural developments, urban expansions, and climate change. The study notes changes in the enzootic territory, affecting mammal migration and consequently altering natural focus boundaries. Industrial activities, rotational methods, and habitat changes contribute to the increased epidemic potential in enzootic areas. Despite anthropogenic and climatic influences, the prevalence of plague remains high in Kazakhstan, with a trend towards expanding enzootic territories. Unified electronic databases on plague, tularemia, anthrax, and other zoonoses, developed for GIS analysis, enable mapping and visualization of natural foci. Electronic maps aid in determining enzootic territory boundaries, assessing infectious disease activity, and planning preventive measures based on risk assessment. ESRI's ArcGIS Desktop 10.8 with Arc Toolbox modules facilitated data processing in the geoinformation environment. Data includes epidemiological examination results, species composition of carriers, and laboratory test outcomes, enhancing comprehensive analysis and decision-making for anti-epidemic measures. The study in Kazakhstan identifies and details six natural and twenty autonomous plague foci, categorizing them by main carriers and observing an expansion of natural hotspots. The enzootic territory is classified into four geographic zones, further divided into 105 landscape-epidemiological regions. Laboratory studies inform electronic maps for analyzing plague's dynamic situation. Anthrax prevalence, primarily in chernozem and chestnut soils, is assessed, revealing 1,778 unaffected settlements and spatially clustered points. An epidemiological index aids in zoning for anthrax trouble. Tularemia's landscape occurrence is classified into four types, with spatial analysis revealing clusters and potential epidemic danger in specific regions. Geographic information technologies highlight high-risk areas, justifying preventive measures for dangerous infections. The results obtained serve as a scientific justification for the priority of preventive measures within the boundaries of administrative territories characterized by a high degree of potential epidemic danger and objectively indicate the prospects for the introduction of GIS technologies into the practice of epidemiological surveillance of particularly dangerous infections.

Evaluation of the expression of isoforms of stem cell factor mRNA in fetal tissues and mononuclear cells at different stages of human development.

We studied the expression of isoforms of stem cells factor mRNA forming as a result of alternative splicing. Both isoforms of stem cell factor mRNA forming as a result of alternative splicing were found in different fetal tissues. Changes in the expression of alternative isoforms of stem cell factor in peripheral blood mononuclear cells were demonstrated from the prenatal and neonatal periods to adult organism.

Ontogenetic features of the expression of mRNA isoforms for leukemia-inhibitory factor in human fetal tissues and mononuclear cells.

The expression of leukemia-inhibitory factor mRNA in human fetal tissues and mononuclear cells was studied during ontogeny. The expression of mRNA isoforms for leukemia-inhibitory factor was tissue-specific at the stage of prenatal development. The transition from antenatal and neonatal development to the postnatal period was accompanied by a decrease in the expression of mRNA isoforms for leukemia-inhibitory factor in mononuclear cells.

Incorporation and localisation of ganglioside GM3 in human intimal atherosclerotic lesions.

Immunohistochemical examination showed that sections of intimal atherosclerotic plaques contained cells and cell clusters as well as areas of extracellular matrix specifically stained with antibodies against ganglioside GM3. No immunohistochemical staining was observed in areas bordering the plaques where there was no histological evidence of atherosclerosis. To determine whether the ganglioside GM3 deposits in the intimal plaques derived directly from plasma or were synthesised by intimal cells. intimal plaque and plasma LDL were assayed for ganglioside GM3 fatty acid composition. This assay showed that more than 50% of the fatty acids of GM3 isolated from both atherosclerotic and normal intima are either minor fatty acids or those absent from LDL GM3. We conclude that the GM3 deposits present in intimal plaque arise in intimal cells and do not derive from plasma LDL.

Products of the reaction of cholesterol with hypochlorite anion.

Products of the reaction of cholesterol with hypochlorite (OCI) in various systems (egg phosphatidylcholine liposomes, low-density lipoproteins, and aqueous colloidal dispersion of cholesterol) were separated and analyzed by TLC, HPLC, and gas chromatography-mass spectrometry. The reactions of hypochlorite with cholesterol result in the same reaction products in all treated systems. Eighteen fractions were isolated from the reaction mixture by normal-phase HPLC in hexane-isopropanol (95:5 v/v); these were then examined by gas chromatography-mass spectrometry. Six products less polar than cholesterol were isolated from the reaction mixture; two of them were identified as 4,6-cholesten-3-one and 4-cholesten-3,6-dione. Among the oxidation products more polar than cholesterol, nine compounds were identified; they are 5,7-cholestadien-3 beta-ol,3,5-cholestadien-7-one, 4-cholesten-3 beta, 6 beta-diol, 5-cholesten-3 beta, 7 beta-diol, cholestan-3 beta, 5 alpha, 6 beta-triol, 5 alpha-cholestan-3 beta-ol-6-one, 5-cholesten-3 beta-ol-7-one, 5 alpha, 6 alpha-epoxycholestan-3 beta-ol, and 5 alpha-cholesten-3,6-dione.

Neutral glycolipids of atherosclerotic plaques and unaffected human aorta tissue.

The composition, structure and localization of neutral glycosphingolipids of human aorta taken from subjects who had died after myocardial infarction were studied. Individual glycosphingolipids were purified by high-performance liquid chromatography and were characterized on the basis of their chromatographic mobility, carbohydrate composition, methylation analysis and by 1H-NMR spectroscopy. The main aortic glycosphingolipids were identified as glucosylceramide, lactosylceramide, globotriaosylceramide and globotetraosylceramide. Significant differences in the neutral glycosphingolipid composition of intima and media were detected. The neutral glycosphingolipid profile of medial plaques resembled that of unaffected media; however, significant differences were detected between intimal plaques and unaffected intima. Whereas the latter contained trihexosylceramide and globoside as the only neutral glycolipids, the intimal plaque glycolipids consisted mainly of glucosylceramide and also contained appreciable amounts of lactosylceramide which were completely absent in the unaffected intima. In comparison to intimal plaques, unaffected intima is characterized by a much higher content of cerebrosides terminating by beta-galactosyl residues which are known to interact with growth factors and other external stimuli. It thus seems possible that the proliferative activity of smooth muscle cells in atherosclerotic diseases is to some extent associated with their neutral glycolipid profile.

The gangliosides of adult human aorta: intima, media and plaque.

The composition, structure and localization of gangliosides of aorta taken from subjects who had died after myocardial infarction were studied. Individual gangliosides were purified by high-performance liquid chromatography and high-performance thin-layer chromatography and were characterized on the basis of their chromatographic mobility, carbohydrate composition, neuraminidase hydrolysis and methylation analysis. The main aortic gangliosides were identified as GM3, GM1, GD3, GD1a and GT1b. Significant differences in the ganglioside composition of intima and media were detected and the ganglioside profile of atherosclerotic plaques was found to differ markedly from that of unaffected intima. The latter was characterized by high content of GD3, a ganglioside thought to be associated with membrane permeability, cell interaction, adhesiveness and growth and to suppress unspecific immune responses. Possible implications of the results in low-density lipoprotein binding to the arterial wall and in immunological changes induced by atherosclerotic lesions are discussed.

Gangliosides of the starfish Aphelasterias japonica, evidence for a new linkage between two N-glycolylneuraminic acid residues through the hydroxy group of the glycolic acid residue.

Mono- and disialogangliosides containing glucose, galactose and sialic acids were isolated from the total lipid extract of hepatopancreas of the starfish Aphelasterias japonica. Their structures were elucidated by total and partial acid hydrolysis, trideuteriomethylation analysis, neuraminidase treatment, chromium trioxide oxidation, methanolysis and periodate oxidation. The monosialoganglioside was identified as 8-O-methyl-N-glycolylneuraminosyl-alpha-(2-3)-galactosyl-beta-(1- 4)-glucosyl-beta-(1-1)-ceramide. The disialoganglioside has the additional N-glycolylneuraminic acid or its 8-O-methyl derivative residue at the subterminal position to which the terminal sialic acid residue is linked through the hydroxy group of the glycolic acid unit. The long-chain bases were found to be mixtures of phytosphingosines with both branched and linear chains, and the fatty acids were shown to be mixtures of normal and alpha-hydroxy fatty acids, the latter amounted to about 90% of the fatty-acid mixtures. The composition of the lipid moieties of the gangliosides was determined by GLC and GLC-MS.

Identification and biological activity of a novel natural prostaglandin, 5,6-dihydro-prostaglandin E3.

A novel natural E-prostaglandin was detected by HPLC among the endogenous prostaglandins extracted from ram seminal vesicles. The corresponding precursor - all-cis-eicosa-8, 11, 14, 17-tetraenoic acid was isolated from bovine liver lipids and the preparative biosynthesis with the microsomal fraction of ram seminal vesicles was performed. The isolated product was purified by HPLC and identified by GC-MS as 5,6-dihydro-PGE3. The results of in vitro tests demonstrate that 5,6-dihydro-PGE3 is 14 times less active uterine stimulant than PGE1, at the same time retaining 75% of the anti-aggregatory potency of PGE1. Thus, 5,6-dihydro-PGE3 meets the requirements of a selective antithrombotic agent more than PGE1.

Molecular species analysis of lipids by metastable ion mass spectrometry.

A convenient universal and fast mass spectrometrical method designed for the molecular species analysis of natural lipids is described. In contrast to the commonly employed procedures the method does not require chemical or enzymatic treatment and does not include chromatographic steps. The method relies on the recognition of ions characteristic of individual molecular species in the mass spectrum of a particular lipid fraction, that is accomplished on the basis of metastable ion spectra. The efficiency of this approach is demonstrated with a variety of natural lipids: triglycerides, glycerophospholipids, sphingomyelin and ornithinolipids. The advantages and limitations of the method as well as possible further developments are discussed.