Analysis of the Morphology of Monocytes and Lymphocytes from COVID-19 Patients Using Low-Voltage Scanning Electronic Microscopy.

Severe course of COVID-19 is largely determined by hyperactivation of the immune system, or cytokine storm, in which immune cells (lymphocytes, monocytes, etc.) play a major role. Using low-voltage scanning electron microscopy, we studied the morphology of lymphocytes and monocytes during cytokine storm. Monocytes and lymphocytes were isolated by fluorescence sorting from the blood of healthy volunteers (n=6) and patients with COVID-19 (n=5) during cytokine storm (IL-6>23 ng/ml, smear positive for SARS-CoV-2). For each patient, 11-32 individual cells were analyzed at magnification of 18-32,000 times. Measurements showed that monocyte size was increased during cytokine storm (p=0.0001).

Development of a Test System to Detect the Omicron Variant of SARS-CoV-2 and the Frequency of Its Detection in Patients.

We developed a new test system to detect the omicron variant of SARS-CoV-2 using allele-specific reverse transcription PCR and estimated the frequency of its detection in patients living in the Novosibirsk Region. Clinical samples were divided into 3 groups: samples collected from December 1 to December 30, 2021 (group 1; n=66), from December 30, 2021 to January 10, 2022 (group 2; n=20), and from January 11 to January 22, 2022 (group 3; n=101). Based on the identification of 5 mutations specific to SARS-CoV-2 (B.1.1.529), two systems of oligonucleotide primers and probes were developed for detecting this coronavirus genotype in clinical samples. Limit of detection (LOD95) was 4×103 genome equivalents per 1 ml of clinical sample for the first test system and 2×103 for the for the second test system. The omicron variant of SARS-CoV-2 was absent in group 1 of studied samples, but was detected in 20% (4/20) of group 2 samples and 88% of group 2 samples collected within less than 2 weeks of January 2022. Using developed test system, we showed that in less than 2 weeks the omicron variant has become dominant in patients, which confirms previously published data on its exceptional contagiousness.

[Characterization of genetic polymorphisms associated with neurophysiological processes and analysis of their allele frequency distribution in the Russian population]. / Kharakteristika geneticheskikh polimorfizmov, svyazannykh s neirofiziologicheskimi protsessami, i analiz raspredeleniya chastoty ikh vstrechaemosti v rossiiskoi populyatsii.

OBJECTIVE: This paper presents an analysis of polymorphic gene loci, the products of which are directly involved in the molecular mechanisms of regulation of neurophysiological processes. MATERIAL AND METHODS: The sample of subjects consisted of 128 unrelated males and females living in the European part of Russia. The study assessed the frequency of occurrence of 11 single-nucleotide substitutions located in genes encoding serotonin receptors, ciliary neurotrophic factor, uncoupling protein 2, methylenetetrahydrofolate reductase, methionine synthase, methionine synthase reductase, dipeptidyl carboxypeptidase 1, gamma-coactivator of the receptor activated by the proliferator peroxisome and neurotrophic factor of the brain. Genotyping of the samples was carried out by PCR with fluorescence detection and analysis of the polymorphism of the lengths of restriction fragments. RESULTS: The distribution of polymorphism genotypes corresponded to the Hardy-Weinberg equilibrium with the exception of rs1801133 MTHFR (χ2=5.3088, p=0.0212) in which a decrease in heterozygosity was observed. These studies of minor allele distribution have no statistically significant deviations from the European population, but there are deviations from the Asian, African, and Latin American populations. CONCLUSION: Statistically significant correlations of allele frequencies in the study group with populations from other regions and the studies conducted in them are the basis for the inclusion of selected single-nucleotide polymorphisms in the list of a limited set of molecular genetic markers, which makes it possible to supplement the system of mental health monitoring and improve the professional training of people in extreme professions.

[Apoptosis and autophagy in the pathogenesis of acute ischemic stroke (review of literature).] / Апоптоз и аутофагия в патогенезе острого ишемического инсульта (обзор литературы).

According to the World Health Organization, about 5 million people die every year from cerebrovascular disease. At the same time, the proportion of cerebral infarction, or ischemic stroke (IS), among forms of acute cerebrovascular accident reaches 80-85%. Despite the active study of biochemical and morphological changes leading to acute cerebrovascular ischemia, the problem of early diagnosis, prevention, as well as predicting the outcome of this disease is still relevant. There is no doubt that the interruption of the ischemic cascade at earlier stages can be accompanied by a greater effect of treatment. A timely and effective pharmacological intervention requires a clear understanding of the pathochemical and biological processes underlying acute ischemia at the molecular level. High mortality and disability accompanying acute IS, dictate the need to create new diagnostic and prognosis algorithms both in the acute period of IS, and in the recovery period. According to some authors, elucidation of the pathways that underlie the pathogenetic mechanisms acting in the penumbra are of great clinical interest for the development of new diagnostic and therapeutic strategies. Studying the mechanisms of apoptosis and autophagy of neurons in the dynamics of the acute period of IS, modulation of the autophagy process in the penumbra zone can contribute to the development of new methods for the diagnosis and treatment of acute IS. The review presents the results of the latest experimental studies on the role of apoptosis and autophagy in the development of acute cerebral ischemia and attempts to modulate these processes in order to influence the ischemic cascade. The review was based on sources from such international and national data bases as Scopus, Web of Science, Springer, RINC.

An Approach to Better Estimation of the Initial Velocity of Enzyme Reactions.

A new algorithm for computation of the initial velocity of enzyme reaction at the time zero is proposed. This algorithm makes it possible to reduce the systematic error of measurements to the minimum, to estimate the reaction velocity in test samples regardless of the enzyme activity levels, and to minimize the assay time. The study is illustrated by an example of salivary alpha-amylase and standard reagent kit. The algorithm should not be applied if conjugated enzyme systems are used because there is a long initial lag phase in the kinetic curve.

[Opportunities of modern laboratory diagnostics of infectious complications of acute pancreatitis (review).]

A systematic search of literary sources in the abstract databases Scopus, Web of Science, MedLine, the Cochrane Library, CyberLeninka, RSCI for 2010-2018. The search queries were: acute pancreatitis and complications, acute pancreatitis and diagnosis, acute pancreatitis and diagnosis and complications, acute pancreatitis and compications, and sepsis. The results of search and analysis of selected literature sources are presented. It was revealed that the currently used set of laboratory and instrumental methods of diagnosis of infectious complications of acute pancreatitis does not fully meet the needs of clinical practice. The most common of them are the determination of blood concentrations Of C-reactive protein and procalcitonin. At the same time, a number of disadvantages of these methods are noted. In the last decade, many new markers of systemic infection have been introduced into clinical practice. Some of them are currently being investigated in order to diagnose systemic infection in General and infectious complications of acute pancreatitis in particular. The most promising are such as presepsin, MID-regional Pro-adrenomedullinum, CD64 neutrophil index and some others.

[The hormonal indices of hypophysis-adrenal glands system in combatants during missions to "trouble spots".]

The officials of the Interior Ministry of Russia usually perform their duties in extreme and quite often emergency conditions. The hypophysis-adrenal glands system of regulation mainly contributes into supporting formation of mechanisms of compensation of these exposures. The purpose of study is to analyze alterations of content of adrenocorticotropic hormone, cortisol, adrenaline and noradrenaline in officials of the Interior Ministry sent on a mission to territories with local military conflict (the Northern Caucasus) to support public order. The study results demonstrated that during first two months of mission a significant increasing of level of analyzed hormones occurs. This is a proper reaction to changed conditions of environment. These alterations correspond to main propositions of the general adaptation syndrome theory. However, further observation in dynamics of mission established presence of signs of imbalance of secretion of hormones in hypophysis-adrenal glands system. The long-lasting conserving high content of both of adrenocorticotropic hormone and cortisol till the end of mission was established. At the same time, content of hormones of "fast response" to extreme factors of environment (adrenaline, noradrenaline) occurred to be as higher as at the beginning of mission. Such occurrences result in disorder of regulation of inter hormonal relationships that correspondingly is a factor of decreasing of vital stress resistance of organism. The main directions of organizational medical character are proposed within th framework of activities supporting resistance of organism to conditions of extreme impact of emergency situations and decreasing risk of development of pathological conditions.

[The clinical laboratory characteristics of secretory function of thyroidal chain of regulation in individuals of dangerous professions].

The established opinions concerning etiology and pathogenesis of human health condition owing to impact of extreme factors indicate that stressing of regulatory systems (neuro-immune endocrinal complex) is a leading mechanism in their development. Just in the endocrine component failure of physiological bindings occurs in the system "central-peripheral internal secretion glands" followed by alteration of metabolism. The study was carried out to determine characteristics of content of levels of thyrotropic hormone, thyroxine and triiodothyronine in individuals of dangerous professions depending on level of implemented occupational load. Two series of studies was implemented among employees of the Interior ministry board - combatants, ordinary policemen and cadets. The examination began with the mission on the territory with armed conflict of combatants and continued after its completion. The blood serum was analyzed for establishing content of thyrotropic hormone, thyroxine and triiodothyronine. The highest level of thyrotropic hormone was detected in group of combatants in both studies. The levels of thyroxine and triiodothyronine were minimal in group of combatants prior to mission as compared with other groups and the highest ones after mission. Therefore, long-term alterations of secretory function of endocrine system resulting in hormonal imbalance can lead to breakdown of adaptation process. Thereupon, the system of medical support of individuals under effect of extreme negative professional factors requires development and adequate functioning of endocrinological component with the purpose of increasing resistance and viability of organism to the conditions of extreme impact of emergency situations and also preventing development of pathological conditions.

[Hypophysis-adrenal and thyroid secretion at law order staff depending on professional loading].

BACKGROUND: A current etiological and pathogenic opinion about human health disturbance thereupon extreme factor effects is shown that this cause is principal mechanism of regulatory system (neuroimmunoendocrine complex) distress. In endocrine link occurs hormonal disbalance in hypothalamus-hypophysis axis, physiological interrelation disturbances in central-peripheral gland system (hypophysis-adrenal, hypophysis-thyroid) and metabolism abnormalities subsequently. OBJECTIVE: Our aim was to determine the particular content of adrenocorticotropic and thyrothrophin hormone, cortisol, thyroxin and triiodthyronine features at law order staff in dependence from professional loading. METHODS: It's provided two investigation series among law order staff groups--combatants, ordinary policemen and military school students. The investigation period for all people corresponds to combat mission beginning and its finish. In blood serum an adrenocorticotropic (ACTH) and thyrothrophin (TSH) hormone, cortisol, thyroxin (T) and triiodthyronine (T) levels were determined. RESULTS: A higher ACTH and TSH levels detected at combatants in both investigation series. A cortisol, T4 and T3 at combatants before military mission were least in comparative with other groups, but after mission it indexes were largest. CONCLUSION: Prolonged changes of endocrine secretory function that lead to hormonal disbalance can result to adaptation derangement. In connection with it in medical providing system for person that undergo extreme negative professional factors it's necessary create a special endocrine link with the view of organism resistance and life viability to extreme emergency factors and for prevention of pathological conditions.

[The clinical laboratory diagnostic as a part of clinical medicine.]

The rapid development of laboratory technologies and their translation directionality permit developing principles of personified medicine. The article considers new paradigm of manpower policy for developing necessary level of clinical laboratory consultation.

[Medical devices and its in-field use].

The quality of in-field treatment and prevention measures depends on the availability of modern medical devices, instruments, and equipment. In this regard peculiarities of medical care delivery during military conflicts advance special requirements for medical equipment, which is used by the medical service of the Armed Forces of the Russian Federation. The article shows prospects of in-field use of the most important samples of medical devices adopted by the Armed Forces.

[Perspective technologies and researches in the areaof medical laboratory diagnostics].

The main principles of organisation of medical laboratory diagnostics are efficiency of analysis, mobility of laboratory services and quality of researches. These goals can be achieved by the use of portative laboratory analizers, by automation and computerization of the laboratorial service, by development and adoption of new laboratory technologies, integrating different methods and types of research. It is necessary to pay attention to the problem of NPT and indication of pathogenic germs. Priority areas of medical laboratory diagnostics development are: development and use of portative laboratory analyzers; development of chemical, that help to speed up and cheapen researches, improve effectiveness of laboratory diagnostics of infections and indications of pathogenic and other germ; development of new, more sensitive, specific, but simple methods of laboratory analysis; development of complex methods and types of researches, further implementation of methods and researches with different principles of action; development and implementation of new methods of NPT results recording; automation and computerization of the laboratorial diagnostics.

[Botulinum toxine treatment and the formation of antibodies to botulotoxin in patients with cerebral palsy].

The aim of the study was to search for the association between efficacy of botulinum treatment (dysport) and the formation of antibodies to botulinum toxin in children with cerebral palsy (CP). Fourteen children with CP received dysport, 42 children (29 with CP and 13 patients with paresises of other etiologies) were not treated with this drug. No association between the presence of antibodies in blood serum and efficacy of treatment was found. However, further research of the role of antibody genesis as a factor influencing the formation of reaction to treatment is needed.

[The effect of 120 day anti-orthostatic hypokinesia on the status of the posture regulation systems]. / Vliianie 120-sutochnoi antiortostaticheskoi gipokinezii na sostoianie sistem poznogo regulirovaniia cheloveka.

The system of posture regulation was evaluated during 120-d head-down bedrest (-6 degrees). Functioning of the system was evaluated by the characteristics of corrective posture responses to pushes in the chest breaking the body equilibrium. Analyzed were parameters of amplitude and time of corrective responses, and strength and range of stimulus intensity. Results of the investigation showed that HD bedrest materially disturbs vertical stability and impairs effectiveness of the posture corrective mechanisms. Based on the data of this investigation and findings of previous efforts we can confidently distinguish two phases in development of disturbances in vertical stability during long-term HD bedrest associated with different leading mechanisms. On the first phase (15-30-d into the experiment), the principle posture problems are reduced tone of the anti-gravity musculature. On the second phase, particularly beginning on day 60, muscular atrophy processes become leading factors of postural disturbances. Apparently, these mechanisms, along with changed vestibular input, considerably impact vertical stability, especially after extended space mission.

[The causes for the worsening of treatment results in patients with infected pseudarthroses of the tibia in recent years]. / Prichiny ukhudsheniia rezul'tatov lecheniia bol'nykh s infitsirovannymi psevdoartrozami bol'shebertsovoi kosti v poslednie gody.

Under analysis were results of treatment of 141 patients with infected pseudoarthroses of the tibia. Among them there were 10 women and 131 men aged from 18 to 62 years. The long-term results were followed-up during the period from 2 to 20 years. The analysis allowed a supposition that a combination of tibia segment resection and plasty with local tissues completed by suturing the wound, drainage and external fixation with the Ilizarov device leads to deterioration of the outcomes. A careful approach to treatment of such patients including the division of treatment into the stages of debridement and reconstruction seems to be more productive.