[Development and preservation of specific T-cell immunity after COVID-19 or vaccination against this infection].

Aim evaluation of specific T-cell immunity against SARS-CoV-2 in primary and secondary response to virus antigens by screening method. MATERIALS AND METHODS: Patients were tested 11.5 months after COVID-19 and 610 months before and after vaccination. Healthy volunteers were screened before, 26 times during the vaccination course, and 68 months after revaccination with the Sputnik V vaccine. IgG and IgM antibodies to SARS-CoV-2 were detected by ELISA using commercially available kits (Vector-Best, Russia). Antigenic (AG) activation of T cells in the fraction of bloods mononuclear cells was assessed by IFN- production after AG stimulation in the wells of plates from ELISA kits intended for detection of antibodies against SARS-CoV-2. Data were processed by MS Excel and Statistica 10.0 software. RESULTS: AG-specific T cells were detected in 88.5% of vaccinated healthy volunteers, half of whom were found to have T cells appearing earlier than antibodies to AG. After 6-8 months, the level of AG activation decreases. Following the revaccination, the level of AG activation of memory T cells in vitro increases within six months in 76.9100.0% of vaccinated subjects. On the contrary, after COVID-19, 86.7% of individuals had in their blood the AG-specific T cells with high activity at the time of vaccination. The activity of T cells recognizing the RBD domain of the SARS-CoV-2 S protein and the proportion of individuals who had these cells in their blood increased after the vaccination of reconvalescents. CONCLUSION: T-cell immunity against SARS-CoV-2 antigens has been shown to persist for 6 months after illness. In vaccinated individuals without history of COVID-19, such duration of the preservation of AG-specific T cells in blood was only achieved after the revaccination.

[Assesment of specific T-cell immunity to SARS-CoV-2 virus antigens in COVID-19 reconvalescents].

INTRODUCTION: The development of the COVID-19 pandemic has stimulated the scientific research aimed at studying of the mechanisms of formation the immunity against SARS-CoV-2. Currently, there is a need to develop a domestic simple and cost-effective specific method suitable for monitoring of T-cell response against SARS-CoV-2 in reconvalescents and vaccinated individuals. AIM: Development of a screening method for evaluation specific T-cell immunity against SARS-CoV-2. MATERIALS AND METHODS: Total 40 individuals who had mild to moderate COVID-19 and 20 healthy volunteers who did not have a history of this disease were examined. The presence and levels of IgG and IgM antibodies to SARS-CoV-2 were identified in participants sera by ELISA using the diagnostic kits from JSC Vector-Best (Novosibirsk, Russian Federation). Antigenic stimulation of mononuclear cells was carried out on commercial plates with adsorbed whole-virion inactivated SARS-CoV-2 antigen (State Research Center of Virology and Biotechnology VECTOR Novosibirsk, Russian Federation). The concentration of IFN- was measured in ELISA using the test systems from JSC Vector-Best (Novosibirsk, Russian Federation). The immunophenotyping of lymphocytes was performed on a flow cytometer Cytomics FC500 (Beckman Coulter, USA). Statistical data processing was carried out using the Microsoft Excel and STATISTICA 10 software package. RESULTS: Stimulation of mononuclear cells isolated from the peripheral blood with whole-virion inactivated SARS-CoV-2 antigen fixed at the bottom of the wells of a polystyrene plate showed a significantly higher median response in terms of IFN- production in 40 people who had history of COVID-19 compared to 20 healthy blood donors (172.1 [34.3575.1] pg/ml versus 15.4 [6.925.8] pg/ml, p 0.0001). There was no difference in median IFN- levels in supernatants collected from unstimulated mononuclear cells from COVID-19 reconvalescents and healthy donors (2.7 [0.411.4] pg/ml versus 0.8 [0.023.3] pg/ml, p 0.05). The overall sensitivity and specificity of this method were 73% (95% CI 5888%) and 100% (95% CI 100100%), respectively, at a cut-off of 50 pg/ml. CONCLUSION: The developed method for assessment of the cellular immune response to SARS-CoV-2 can be used as a screening method for monitoring the T-cell response in a population against a new coronavirus infection in recovered people.

[Progressive fibrosing lung disease. Discussion aspects of the problem: A review].

The authors of the article prove the need to include a new name for the disease - "Progressive Fibrosing Lung Disease" into clinical practice. Recognition of the fact that some lung diseases end in a fibrosing process, which does not have any significant differences depending on the initial disease that led to fibrosis, will expand the indications for earlier prescription of antifibrotic drugs, which will undoubtedly improve the prognosis in this extremely severe category of patients.

Alkaloid content variability in the seeds of narrow-leafed lupine accessions from the VIR collection under the conditions of the Russian Northwest.

Alkaloid content was assessed in the seeds of 59 narrow-leafed lupine (Lupinus angustifolius L.) accessions from the VIR collection in the environments of Leningrad Province. The selected set included accessions of different statuses (wild forms, landraces, and advanced cultivars) and different years of introduction to the collection. Alkaloids were analyzed using gas-liquid chromatography coupled with mass spectrometry. Concentrations of main alkaloids: lupanine, 13-hydroxylupanine, sparteine, angustifoline and isolupanine, and their total content were measured. The total alkaloid content variability identified in the seeds of the studied set of accessions was 0.0015 to 2.017 %. In most cases, the value of the character corresponded to the accession's status: modern improved cultivars, with the exception of green manure ones, entered the group with the range of 0.0015-0.052 %, while landraces and wild forms showed values from 0.057 to 2.17 %. It is meaningful that the second group mainly included accessions that came to the collection before the 1950s, i. e., before the times when low-alkaloid cultivars were intensively developed. Strong variability of the character across the years was observed in the accessions grown under the same soil and climate conditions in both years. In 2019, the average content of alkaloids in the sampled set was 1.9 times higher than in 2020. An analysis of weather conditions suggested that the decrease in alkaloid content occurred due to a significant increase in total rainfall in 2020. Searching for links between the content of alkaloids and the type of pod (spontaneously non-dehiscent, or cultivated, spontaneously dehiscent, or wild, and intermediate) showed a tendency towards higher (approximately twofold in both years of research) total alkaloid content in the accessions with the wild pod type and the nearest intermediate one compared to those with the pod non-dehiscent without threshing. The correlation between the average total alkaloid content and seed color, reduced to three categories (dark, or wild, light, or cultivated, and intermediate), was significantly stronger in the group with dark seeds (5.2 times in 2019, and 3.7 times in 2020). There were no significant differences in the percentage of individual alkaloids within the total amount either between the years of research or among the groups with different pod types or the groups with different seed coat colors.

[Predictors of lethal outcome in young patients with hemorrhagic stroke]. / Prediktory letal'nogo iskhoda gemorragicheskogo insul'ta u patsientov molodogo vozrasta.

OBJECTIVE: To evaluate the predictors of the lethal outcome of hemorrhagic stroke in young people. MATERIAL AND METHODS: A retrospective analysis of cases of non-traumatic hemorrhagic stroke was carried out in 60 young patients hospitalized in the neurosurgical department in the Ufa Clinical Emergency Hospital. By gender, the male predominated. The mean age of the patients was 37.4±5.1 years. Lethal outcome was in 18 (30%) patients. RESULTS: Data analysis revealed a relationship between the lethal outcome of hemorrhagic stroke and certain clinical, somatic, laboratory and instrumental factors. As a result, a significant model (p<0.001) of the probability of a lethal outcome in young patients was developed. CONCLUSION: The identified predictors of the probability of death from stroke can serve as the basis for adequate management of young patients with hemorrhagic stroke.

Phenotypic traits differentiating the genetic resources of pea (Pisum sativum L.) by the type of use.

The paper presents an analysis of the data obtained for pea accessions from the VIR collection studied at the Adler Experiment Station in the setting of the Krasnodar Territory in 2017-2019. It was for the f irst time that these accessions were studied for a set of phenotypic traits. The object of the study was a sample of 494 pea accessions originated from 43 countries and 18 regions and territories of the Russian Federation. The work was carried out in compliance with the methodological guidelines developed at VIR. Statistica 13.3 software was employed for statistical data processing. An assessment of four qualitative, 10 quantitative and four phenological traits in the accessions made it possible to differentiate them by the type of use, that is, as dry, forage and garden peas. The varieties differing in the type of use signif icantly differed by the values of such traits as stem length, number of pods per plant, number of nodes to the f irst f lower, number of f lowers in the inf lorescence, the maximum number of seeds per pod, pod length, and a narrower pod of forage pea compared to that of dry and garden peas. The average values of these traits were recorded for the peas with different types of use. The maximum difference was noted between garden and forage pea varieties. Dry pea varieties occupied an intermediate position. The complex of phenotypic traits identif ied determines the differences between three types of pea use, which is important when selecting the initial material for breeding appropriate varieties.

[Ethiopathogenetic features and course of ischemic stroke associated with COVID-19]. / Etiopatogeneticheskie osobennosti i techenie ishemicheskogo insul'ta, assotsiirovannogo s COVID-19.

OBJECTIVE: To analyze etiopathogenetic factors and course of ischemic stroke associated with new coronavirus infection (COVID-19). MATERIAL AND METHODS: A retrospective clinical study of 173 patients with ischemic stroke and COVID-19 (main group) and 86 patients with ischemic stroke without COVID-19 (comparison group) was carried out. There were no statistically significant differences in age and gender. All patients underwent standard clinical-instrumental, laboratory and neuroimaging assessments. RESULTS: Compared with the comparison group, patients with COVID-19 were less likely to have cardiovascular risk factors, the difference being statistically significant. Stroke in the main group was more severe than in the comparison group. According to the TOAST classification, an unknown stroke subtype significantly predominated in the main group. Laboratory data in the main group indicated the significance of an increase in renal-hepatic markers (creatinine, aspartate aminotransferase) and systemic inflammatory response syndrome (C-reactive protein). CONCLUSION: The development of cardiovascular diseases in patients with COVID-19 is an important negative prognostic factor that requires further study to determine the optimal management strategy.

Features of the Humoral Response to Infection, Vaccination, and Revaccination during COVID-19.

IgM and IgG antibodies to the SARS-CoV-2 virus are detected in subjects who have recovered from COVID-19; IgM antibodies persist in a 1/3 of infected subjects up to 12 months from the moment of the disease, while IgG antibodies are present in the vast majority of cases (97%; medium and high levels antibodies were registered in 85% of cases). By the 12th month, 40% of those who recovered still have a very high level of IgG antibodies to the S-protein (>500 BAU/ml). In the feces, urine, and blood serum of patients with long-term persistent IgM antibodies, no coronavirus antigens were detected. After vaccination with the Gam-COVID-Vac vaccine, IgG antibodies to the S-protein are detected in 100% of cases and remain at a high level for 4 months, by the 5-6th month, the level of antibodies decreases. During revaccination, the level of IgG antibodies to S-protein reaches high values earlier than during primary vaccination, and remains high for 4 months (observation period). The blood sera of recovered and vaccinated patients have a high virus-neutralizing activity (at least 1:80), while its level is somewhat higher in recovered patients.

[Dissection of cerebral arteries as a cause of ischemic stroke]. / Dissektsiya tserebral'nykh arterii kak prichina ishemicheskogo insul'ta.

Dissection of cerebral arteries is a rare cause of stroke in the general population. The article describes 5 clinical cases of patients with ischemic stroke after dissection, discusses the issues of etiology, diagnosis and treatment tactics. The possibility of using high-tech methods of treatment for this subtype of stroke is emphasized. The diagnostic method of choice is neuroimaging, including cerebral angiography.

[Therapy of moderate cognitive impairment and asthenia in patients with cerebrovascular pathology: results of a prospective observational study]. / Terapiya umerennykh kognitivnykh narushenii i astenii u patsientov s tserebrovaskulyarnoi patologiei: rezul'taty mnogotsentrovoi otkrytoi prospektivnoi nablyudatel'noi programmy.

OBJECTIVE: Obtaining additional data on the efficacy and safety of the drug Prospekta in the treatment of moderate cognitive impairment (MCI) and asthenia in patients with cerebrovascular disease (CVD). MATERIAL AND METHODS: A prospective observational study in more than 40 Russian cities enrolled 232 patients (mean age 61.5±10.0 years) with mild cognitive impairment (MCI), asthenia on ongoing basic nootropic therapy. The presence of MCI was confirmed by the Montreal Cognitive Assessment Scale (MoCA), asthenia - by 10-point Visual Analog Scale (VAS). All patients were prescribed the nootropic medication Prospekta 2 tablets 2 times a day for 8 weeks in addition to the therapy they received. Ultrasound Doppler sonography of the main arteries of the head and magnetic resonance imaging (MRI) of the brain were also assessed. At the end of treatment, the Clinical Global Impression Efficacy Index (CGI-EI) was assessed and the safety of the treatment was evaluated. RESULTS: The baseline severity of cognitive impairment according to the MoCA scale was 21.6 points, severity of asthenia according to the VAS was 6.3 points. According to Doppler flowmetry findings, hemodynamically significant stenosis was revealed in 105 (49.3%) patients, and narrowing of the main vessels without changes in hemodynamic parameters was revealed in 108 (50.7%) patients. According to MRI results, single vascular lesions in the brain matter were detected in 102 (44.0%) patients. The medications with nootropic effect were administered to 144 (62.1%) patients. A positive therapeutic response as improvement of cognitive functions was seen in 93.3% of patients after 8 weeks of taking Prospekta, including 39.4% of patients who had cognitive functions restored to the normal level. No side effects were registered during the observational study. CONCLUSIONS: The nootropic medication Prospekta is effective and safe in treatment of MCI in patients with asthenia with CVD, and improves cognitive function in patients with asthenia with CVD, both in monotherapy and in combination with other nootropic agents.

[Predictors of unfavorable outcomes of ischemic stroke associated with COVID-19]. / Prediktory neblagopriyatnogo iskhoda ishemicheskogo insul'ta, assotsiirovannogo s COVID-19.

OBJECTIVE: To study predictors of unfavorable outcomes in ischemic stroke associated with COVID-19. MATERIAL AND METHODS: A retrospective analysis of 173 case histories of patients with ischemic stroke and COVID-19 was carried out. The average age was 68.64±11.39 (95% CI 66.93-70.35; median 92, mean 34) years. By gender, women predominated (64.16%). Fatal outcome was observed in 62 (35.84%) patients. Risk factors were studied using binary logistic regression. RESULTS: The risk of a fatal stroke in a univariate analysis was established in the presence of a general severe condition of the patient and a number of other factors, which included the severity of COVID-19, acute coronary syndrome, multiple organ failure, the need for a ventilator, a history of kidney disease, pneumonia, high NIHSS score, oxygen partial pressure level, respiratory rate, number of hospitalizations, complete blood count (erythrocytes, hemoglobin, hematocrit, leukocytes, neutrophils), coagulogram, glucose, liver and kidney markers (bilirubin, aspartate aminotransferase, alanine aminotransferase, creatinine, urea), levels of creatine phosphokinase, lactate dehydrogenase and C-reactive protein. A model based on multivariate analysis was used, which makes it possible to predict the probability of a fatal outcome. A regression function was obtained, which included the content of C-reactive protein, urea and NIHSS score. In patients with a value of 35% and above, there was an increased risk of death, in the case of a value below 35%, a favorable outcome was assumed. The model was statistically significant (p<0.001). The sensitivity and specificity of the model were 88.9% and 97.9%, respectively. CONCLUSION: The revealed predictors of the probability of a fatal stroke outcome can be guidelines for a Coctor in choosing a patient management strategy at different stages of patient care.

Forecast for the zone of viticulture in European Russia under climate change.

Climate warming has turned out to be a significant factor in viticulture and winemaking in all grape-growing areas of the world. Many countries consider the advance of viticulture to the north and to mountainous areas as a possible way to adapt to warming. The factors limiting the zone of viticulture in Russia have been identified by Soviet scientist F.F. Davitaya in 1948, and they are still relevant. They are the sum of active temperatures above 10 °C (ΣT10 > 2500 °C), mean of absolute minimum temperatures (Tmin > -35 °C), length of the frost-free period (Lff < 150 days), and hydrothermal coefficient (0.5 < HTC <2.5). The values of these limiting factors in the present-day zone of commercial viticulture (ZCV) correspond to the ranges defined by F.F. Davitaya, with the exception of Tmin, which in the modern ZCV in European Russia is above -26 °C everywhere. The objective of this work was to assess the possibility of moving the boundaries of the ZCV to the north under the existing and predicted climate conditions in European Russia. The 1980-2019 daily data from 150 weather stations of the Federal Service for Hydrometeorology and Environmental Monitoring were used to calculate mean long-term values, trends and forecasts for 2050 for the ZCV limiting factors and locate the points lying in the range acceptable for viticulture. The QGIS program was applied to plot the points on the European Russia map and mark the terminal latitude. Versions with Tmin > -26 °C and Tmin > -35 °C were considered. On average for European Russia, in 1980-2019, there was an increase in ΣT10 , Tmin, and Lff and a decrease in HTC. However, in the same period, Tmin showed a tendency toward decreasing at a number of points at latitudes lower than 55° N. The increase in heat supply during the growing season in European Russia implies a possibility of expanding the ZCV northward, beyond the present-day terminal latitude of 46.6° N, to 51.8° N under the existing conditions, and up to 60.7° N by 2050. In addition, even under the current conditions viticulture is possible in the area of Kaliningrad (54° N, 20° E). Using extra protective measures in winters not colder than -35 °C would make it possible to grow grapes at up to 53.3° N under the current conditions and at up to 60.7° N under the prognosticated ones. At the same time, a possible decrease in the minimum winter temperature at the south of European Russia will require additional protective measures in winter, while an increase in the aridity of the climate on the northwest coast of the Caspian Sea will reduce the area under non-irrigated vineyards.

[Pulmonary epithelioid hemangioendothelioma]. / Epitelioidnaya gemangioendotelioma legkikh.

The article describes the clinical, radiological and pathological features of epithelioid hemangioendothelioma (EHE) in 27 adult patients, mainly female. In all cases, with the exception of one, there was a benign course of the disease over many years with a tendency to stabilize growth, the morphological sign of which was the development of widespread sclerotic changes. With the help of immunohistochemical method, the endothelial nature of EHE cells and its relatively low proliferative potential were confirmed. Clinical and morphological features of EHE raise the question of the essence of proliferation of endothelial cells with the formation of tumor-like nodes. There is every reason to consider EHE as a pseudotumor of the type of nodose hyperplasia in the nosological group of dyshormonal hyperplasia, similar to benign leiomyoma of the uterus with lung damage, as we have previously proposed.

[Etiopathogenetic features, course and outcome of hemorrhagic stroke in young people]. / Etiopatogeneticheskie osobennosti, techenie i iskhod gemorragicheskogo insul'ta u lits molodogo vozrasta.

OBJECTIVE: Analysis of etiopathogenetic factors and features of the course of hemorrhagic stroke in young people. MATERIAL AND METHODS: A retrospective analysis of 402 case histories with non-traumatic hemorrhagic stroke was carried out, of which 60 (14.93%) patients were young. The average age was 37.37±5.09 years. There were 19 (31.7%) females and 41 (68.3%) males. Statistical data processing was carried out using Microsoft Office 2010 and SPSS v.26. RESULTS: The main causes of hemorrhagic stroke in young people were changes in cerebral vessels by the type of aneurysmal deformation of the vascular wall, including Moya-Moya disease, and arteriovenous malformations (60%). The second etiological factor of hemorrhagic stroke was arterial hypertension (40%). Intracerebral hematoma was the most common type of hemorrhagic stroke. The functional outcome was influenced by: the severity of the patient's general condition, the severity of the neurological deficit, the degree of consciousness, the presence of concomitant pathology, convulsive, dislocation syndrome, leukocytosis, hyperglycemia, high creatinine levels. CONCLUSION: The problem of hemorrhagic stroke at a young age is complex and relevant, since the etiological factor of the disease at this age differs significantly from that in older age groups, requires adequate diagnostic searches to identify the etiopathogenetic factor and subsequent secondary prevention.

Dynamics of Antibodies to Various Antigens of the SARS-CoV-2 Coronavirus in Patients with Confirmed COVID-19 Infection.

The presence of IgG and IgM antibodies in the venous blood of 76 patients with confirmed COVID-19 infection was determined by ELISA using Russian test systems. Different levels of IgM antibodies to N-protein and receptor binding domain of the Spike protein (RBD) were revealed. The dynamics of IgG antibodies to the whole virion antigen and recombinant antigens showed high values on weeks 4-5 of the disease. The level of IgG antibodies to Nprotein remained low throughout the observation period. The characteristic dynamics of IgG measured using test systems with sorbed whole virion or recombinant spike proteins reflects the duration of the disease.

[Idiopathic interstitial pneumonias].

The article presents the evolution of views on one of the current problems of present pulmonology idiopathic interstitial pneumonias. On the basis of many years of experience in diagnosis and treatment of patients of IIPs in the clinic of pulmonology of Interstitial and Orphan Lung Diseases Research Institute of Pavlov First Saint Petersburg State Medical University, the authors formulated a new understanding of this pathology and proposed to unite all IIPs under the term idiopathic fibrosing pulmonary disease. Using the concept of idiopathic fibrosing pulmonary disease will make it possible to substantively address the issues of early diagnosis, determine the criteria for the activity of the pathological process, and there by develop an evidence base for the adequate prescription of antifibrotic drugs and corticosteroids.

[Factors affecting the effectiveness of rehabilitation in patients in the acute period of cerebral stroke]. / Faktory, vliyayushchie na effektivnost' reabilitatsii bol'nykh v ostrom periode tserebral'nogo insul'ta.

AIM: To study the factors, affecting the effectiveness of acute period rehabilitation in patients after ischemic stroke (IS). MATERIAL AND METHODS: We examined 72 patients (average age 63.8 ± 1.3 years) in the acute period of IS who were treated in the neurological department for patients with acute cerebrovascular accident of the State Budgetary Institution of Healthcare of the Republic of Bashkortostan «Emergency Hospital¼ (Ufa). All patients underwent clinical, instrumental and laboratory examination. The following scales were used: NIHSS, Rankin, Barthel, Rivermead, Montreal Cognitive Assessment, Spielberg - Hanin situational and personal anxiety scale, Beck's Depression Inventory, A.M. Vain questionnaire for assessment of the severity of autonomic dysfunction, T. Ehlers test and A.M. Schubert's method of determination of the state of the motivational sphere and risk preparedness. Assessment of the rehabilitation potential was carried out by analyzing the data of the Rehabilitation List. According to the severity of neurological deficit, the patients were divided into 2 groups: 38 (52.7%) patients with mild IS were included in the Group 1 and 34 (47.2%) patients with moderate IS - in Group 2. RESULTS: By the end of the treatment, significant changes in the number of patients with cognitive and depressive disorders, qualitative changes in the levels of personal and situational anxiety in the direction of their reduction were registered. In the vast majority of patients in both groups at the stages of treatment, one or another degree of autonomic nervous system dysfunction was found. Most patients (82.9%) at the beginning of the treatment had an average and high degree of motivation to achieve success in treatment, along with a low and medium level of risk preparedness, which is characteristic of a person aimed at success. According to the complex of factors, 78% of patients had an average and high level of rehabilitation potential. CONCLUSION: A comprehensive and individual approach to the correction of pathological disorders in patients in the acute period of IS, taking into account the rehabilitation potential, is the key to the effectiveness of rehabilitation treatment at the stages of rehabilitation.

Analysis of the Pharmacokinetics of Suppository Forms of Bacteriophages.

Pharmacokinetics of suppository forms of bacteriophages was studied on male Chinchilla rabbits. Suppositories with various composition of bacteriophages were administered once per rectum to rabbits, and the presence of phage particles was estimated in the blood, urine, and feces over 24 h. Pharmacokinetic study showed that the phages were detected in the blood, urine, and feces at various terms of the experiment irrespective of the size of viral particles, which confirmed the possibility of their systemic effects after rectal administration. Thus, the use of suppository form of bacteriophages can ensure the presence of phage particles even in infection foci that cannot directly contact with the preparation.

[Vascular remodeling with violations of intracardiac hemodynamics in patients older age category, combined with the clinical-cluster, neurocognitive and biomarker heterogeneity in multifocal atherosclerosis].

AIM: Study of the remodeling of the carotid arteries with violation of intracardiac hemodynamics in patients with MFA, and the estimation of the main parameters of dyslipidemia, apoptosis, and oxidative stress in patients with high vascular risk older age group (6175 years) in a Regional vascular center of Ufa. MATERIALS AND METHODS: Depending on the predominant lesion of the vascular pool, patients were divided into 3 clusters by the method of hierarchical analysis of categorical variables according to the clinical manifestation of atherosclerotic lesions of the heart, brain and lower limb arteries confirmed by coronary angiography, ultrasound Doppler of the main arteries of the head and lower extremities. 96 of them were IPA with a primary lesion of the heart (1st cluster), the 96 IPA with a predominance of lesions of the carotid arteries (2nd cluster), 96 patients with ischemia of lower extremities (3rd cluster). At the hospital stage, electrocardiography, echocardiography, magnetic resonance imaging of the chest and abdomen, ultrasound of the OBP and kidneys, if necessary, ultrasound of the pelvis were performed. Determination of 8-ON-deoxyguanosine, annexin-5 (An-5) and Aan-5 in blood by ELISA was performed in all patients with MFA, as well as standard biochemical screening for lipidogram examination. RESULTS: We have found that most often in different combinations and with different degrees of severity according to our data are observed: Clinical manifestation of atherosclerotic heart disease (cluster 1) mainly due to its history in combination with stage III hypertension with increasing thickness of intima-media complex and stenosis of the right WASP, left ventricular dilatation, as well as a higher concentration of Aan-5IgMand LP-A as a risk factor for coronary heart disease, atherosclerosis, atherothrombosis. 2. Hemodynamically significant violations of the main arteries of the head in patients of the 2nd cluster mainly with acute ischemic cerebral circulation, in which there was a development of left ventricular hypertrophy with an increase in the size of the left atrium and the presence of atherosclerotic plaque of the right and left WASP. The higher prevalence of stroke was combined with a marked cognitive deficit among patients of cluster 2 with the lowest level of An-5, an increase in total cholesterol and low-density lipoprotein cholesterol. 3. The total severity of the condition in patients with hemodynamic ischemia with clinical manifestation of vascular lesions of the lower extremities was accompanied by a predominant increase in stable angina with FC2, lerish syndrome with occlusion of the iliac, superficial femoral arteries, the presence of insulin-independent type 2 diabetes, which in this group was established in 59.4% of cases, combined with a higher concentration of the marker of oxidative stress 8-ON-deoxyguanosine and hypertriglyceridemia. CONCLUSION: The construction of a three-cluster model in patients at high vascular risk of the elderly age category showed the interaction of cardio-carotid comorbid background on the clinical diversity of systemic vascular lesions in MFA with the development of remodeling of the main arteries and disorders of intracardiac hemodynamics associated with laboratory changes in the assessment of the main parameters of dyslipidemia, apoptosis markers, oxidative stress.

Analysis of In Vivo Activity of the Bovine Cholesterol Hydroxylase/Lyase System Proteins Expressed in Escherichia coli.

The cholesterol hydroxylase/lyase (CHL) system, located in the mitochondria of the mammalian adrenal cortex cells, consists of cytochrome P450scc (CYP11A1), adrenodoxin (Adx), and adrenodoxin reductase (AdR) and performs the first stage of the steroidogenesis: AdR and Adx enable the electron transfer between NADPH and cytochrome P450scc, and P450scc catalyzes the conversion of cholesterol into pregnenolone. CHL system was reconstructed in Escherichia coli using the polycistronic plasmid pTrc99A/CHL. In E. coli cells, the recombinant proteins form the catalytically active system. CHL activity towards 22R-hydroxycholesterol was 4.0 ± 1.3 nmol pregnenolone/h per 1 mg homogenate protein. The alteration of the order of heterologous cDNAs in the expression cassette from AdR-Adx-P450scc to P450scc-Adx-AdR results in alteration of stoichiometric ratio P450scc/Adx/AdR from 1:1.45:4.2 to 1:1.67:0.98; the former ratio is more optimal for the functioning of the cytochrome P450scc. The application of modified cDNA of Adx (AdxS112W) does not increase the CHL activity; however, the introduction of the second copy of AdxS112W gene into the expression cassette increases both the expression level of АdxS112W and the CHL activity in comparison with P450scc/АdxS112W/AdR system. In vivo activity of the CHL system in bacteria is limited by the substrate uptake by bacterial cells: it varied in the range of 0.05-0.62 mg pregnenolone/l resting cell suspension per 1-day cultivation, depending on the type and concentration of permeabilizing agents in the medium. The obtained results contribute to the knowledge of CHL system functioning in living bacteria.