[Evaluation of the dynamics of detection of viable SARS-CoV-2 (Coronaviridae: Betacoronavirus: Sarbecovirus) in biological samples obtained from patients with COVID-19 in a health care setting, as one of the indicators of the infectivity of the virus].

INTRODUCTION: The study of the mechanisms of transmission of the SARS-CoV-2 virus is the basis for building a strategy for anti-epidemic measures in the context of the COVID-19 pandemic. Understanding in what time frame a patient can spread SARS-CoV-2 is just as important as knowing the transmission mechanisms themselves. This information is necessary to develop effective measures to prevent infection by breaking the chains of transmission of the virus. The aim of the work is to identify the infectious SARS-CoV-2 virus in patient samples in the course of the disease and to determine the duration of virus shedding in patients with varying severity of COVID-19. MATERIALS AND METHODS: In patients included in the study, biomaterial (nasopharyngeal swabs) was subjected to analysis by quantitative RT-PCR and virological determination of infectivity of the virus. RESULTS: We have determined the timeframe of maintaining the infectivity of the virus in patients hospitalized with severe and moderate COVID-19. Based on the results of the study, we made an analysis of the relationship between the amount of detected SARS-CoV-2 RNA and the infectivity of the virus in vitro in patients with COVID-19. The median time of the infectious virus shedding was 8 days. In addition, a comparative analysis of different protocols for the detection of the viral RNA in relation to the identification of the infectious virus was carried out. CONCLUSION: The obtained data make it possible to assess the dynamics of SARS-CoV-2 detection and viral load in patients with COVID-19 and indicate the significance of these parameters for the subsequent spread of the virus and the organization of preventive measures.

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.

Determination of Bactericidal Activity Spectrum of Recombinant Endolysins of ECD7, Am24, Ap22, Si3, and St11 Bacteriophages.

The bactericidal activity of recombinant endolysins LysECD7, LysAm24, LysAp22, LysSi3 and LysSt11 was assayed in multidrug resistant strains (n=120) of Salmonella enterica, E. coli, Acinetobacter baumannii, Enterobacter spp., Pseudomonas aeruginosa, Klebsiella pneumoniae, and Campylobacter jejuni. The assay showed that the recombinant endolysins had a wide spectrum of bactericidal activity compared to endolysins of their progenitor phages. Among examined endolysins, we selected the active pharmaceutical substances with broad spectrum of bactericidal activity. Most strains were sensitive to LysECD7 (70.7%), LysAm24 (65%), and LysAp22 (58.6%), which seems to be promising causative agents for the development of finished dosage form.

Establishing of Peritoneal Surface Malignancy Treatment Program: Background in Ukraine, Patient Selection, Staff Schedule, Perioperative Outcomes of First 28 Patients.

A combination of cytoreductive intervention with the intrahepatic hyperthermic chemotherapy is a well-established treatment of peritoneal carcinomatosis. There are many challenges in establishing a new peritoneal surface malignancy program in a low-income country regarding perioperative safety and resource allocation. We analyzed the retrospective database to determine the immediate surgical outcomes of the first 28 patients with peritoneal carcinomatosis and the impact of the program on allocation of general hospital and human resources. During the preparatory stage, web conferences with international experts established the goals and general requirements of the program, formalized in a written document; and a peritoneal surface malignancy multidisciplinary team was established. The team created the patient selection criteria, perioperative management checklists, and suggested material and staff requirements for surgical and anesthesia services. Twenty-nine cytoreduction were carried out, of which 18 were followed by HIPEC. The average operative time was 6.2 h. The median blood loss was 350 mL, which did not result in additional blood transfusion requirements for the blood bank. No additional nursing and supportive staff were provided for the ICU and the surgical unit. The average postoperative stay was 8.7 days. One patient died with 30-day mortality rate of 3%. Ten patients (34.5%) developed postoperative complications, of which 6 (20.6%) had grade III and higher Clavien-Dindo complications. Establishing a peritoneal surface malignancy program is possible via staged planning, liberal use of international mentors, and thoughtful resource allocation. This new program did not result in a significant redistribution of resources among other service lines of cancer care within the same institution.

[Influence of coagulated egg melange on the physical endurance of growing male Wistar rats: physiological and biochemical assessment].

Melange, is one of the most commonly used products of egg processing, however, its use for the production of functional foods is limited by its liquid consistency. The range of functional foods, including those that increase physical endurance and working ability, can be expanded by using the coagulated form of the melange. Thus, the preclinical testing of the nutritional value of coagulated melange and the effect of its consumption on the physical endurance of laboratory animals is relevant. The aim of this research was the experimental comparative physiological and biochemical in vivo evaluation of the effects of coagulated melange consumption by Wistar male rats on their growth indices and physical endurance. Material and methods. Samples of the lyophilized egg melange (EM) and coagulated egg melange (CEM) were used as objects of the research The experiment was conducted with the use of 45 growing male Wistar rats with initial body weight 80±5 g. According to the preliminary training results, 27 animals were selected for further experiment. The motor function was evaluated by means of grip test. The rats were randomized into 2 experimental groups according to their weight and grip strength. Animals were treated for 32 days with isocaloric and isonitrogenic semi-synthetic diets with EM and CEM as protein component (25% of the dry weight), consequently. The grip test was conducted once per week, two days a week animals were trained on the treadmill for 10 min. On the 31st day of the experiment, animals were exposed to exhaustive training till total exhaustion. On the 32nd day of experiment animals were decapitated under light ether anesthesia. The concentration of glucose, triglycerides, cholesterol (CHL), high-density lipoproteins (HDL) and low-density lipoproteins (LDL) was determined in the blood serum. Results and discussion. The greater consumption of food (20±1 g/rat/day) containing CEM by animals corresponded to a greater increase (204±9%) of their body weight over the entire experiment period compared with animals that received EM (13±1 g/rat/day; 160±9%, respectively, p≤0.05). An increase in the intake of food containing CEM had a pronounced positive effect on the physical endurance of the animals, assessed in the grip test. For the 21 days of the experiment, this indicator increased by 26% in rats consuming EM, whereas in rats consuming CEM, the grip strength increased more significantly - by 42%. Consumption of CEM by animals increased the run time (33.8±1.4 min) and the distance covered (557±35 m) in the treadmill test before the exhaustion in comparison with similar indicators for animals that received EM (24.8±2.5 min and 365±50 m, respectively , p≤0.05). The obtained results indicate an increased resistance of animals treated with CEM to exhausting physical loads. Conclusion. The consumption of CEM by growing Wistar male rats increased more effectively their growth and physical endurance in comparison with animals treated with non-coagulated egg melange. These results show the prospective of the use of CEM as a functional food ingredient in specialized foods, enhancing physical endurance and working ability.

Guide RNA Design for CRISPR/Cas9-Mediated Potato Genome Editing.

The activity of the pool of sgRNA molecules designed for different regions of potato coilin and phytoene desaturase genes was compared in vitro. Due to the presence of nucleotides unpaired with DNA, sgRNA is able not only to inhibit but also to stimulate the activity of the Cas9-sgRNA complex in vitro. Although the first six nucleotides located in the DNA substrate proximally to the PAM site at the 3' end are the binding sites for cas9, they had no significant effect on the activity of the Cas9-sgRNA complex.

[The evaluation of genotoxic effects in buccal epithelium under disorders of adaption status of organism.]

The cyto-morphologic analysis of buccal epithelium is one of technique of evaluation of adaptation status of organism and xenogeneic intoxication. The analysis of reactivity of cells of buccal epithelium is used to obtain information concerning genetic alterations in human cells that is extremely important for proper decision making related to issues of population health protection. The smoking results in manifestation of tension of adaptation mechanisms and significant increasing of risk of development cytogenetic disorders in cells of buccal epithelium of oral cavity. The study was carried out concerning morphology of buccal epithelial cells of smoking youths. The cytogram demonstrated a reliable decreasing of number of normal cells and increasing of percentage of cells with various cytogenetic alterations. The cells with micro-nuclei are observed twice more often that can be considered as an integral indicator of genetic disorders in inter phase. In smokers, among cells with signs of completion of nucleus destruction the cells with karyorrhexis are registered reliably more often. The results of micro-nucleus test are an unfavorable prognostic indication testifying high degree of genotoxicity of tobacco smoke and meaning a disorder of cytogenetic homeostasis and decreasing of adaptation reserve of organism.

Multi-subunit BCG booster vaccine GamTBvac: Assessment of immunogenicity and protective efficacy in murine and guinea pig TB models.

New innovative vaccines are highly needed to combat the global threat posed by tuberculosis. Efficient components-antigens and adjuvants-are crucial for development of modern recombinant TB vaccines. This study describes a new vaccine (GamTBvac) consisting of two mycobacterial antigen fusions (Ag85A and ESAT6-CFP10)-with dextran-binding domain immobilized on dextran and mixed with an adjuvant consisting of DEAE-dextran core, and with CpG oligodeoxynucleotides (TLR9 agonists). GamTBvac and its components were assessed for immunogenicity and protective efficacy in GamTBvac-prime/boost and BCG-prime/ GamTBvac-boost in murine and guinea pig TB models. Results show that in both infectious models, GamTBvac has a strong immunogenicity and significant protective effect against Mycobacterium tuberculosis strain H37Rv under aerosol and intravenous challenges. GamTBvac showed a particularly strong protective effect as a BCG booster vaccine.

Searching for active mobile genetic elements in dsRNA fraction of Pinus sylvestris having witches broom abnormalities.

The most common type of coniferous mobile genetic elements are retrotransposons. Despite of their early positive impact on evolution of modern coniferous species they can have a significant negative impact for Forestry and breeding. Breaking genomic structural integrity mobile elements can cause phenotypic defects of plants. In this regard, the study of the diversity of coniferous mobile genetic elements is particularly interesting. In the present paper, we describe mobile genetic elements in dsRNA fraction of Pinus sylvestris having witches broom abnormalities. In result of assembled contigs analysis by RepeatMasker 70 mobile genetic elements were identified. A 68 of that were retroelements. Most of elements represented by Gypsy (16 contigs) and Copia (48 contigs). In 4 cases retroelements specific to Pinus taeda were identified. In most cases fragments of integrase (24), reverse transcriptase (22) and RNaseH (15) were identified. Results of the study may be of interest for coniferous breeding and genetic specialists. The raw data of these experiments have been deposited at NCBI under the accession number SAMN06185845.

A conserved region in the Closterovirus 1a polyprotein drives extensive remodeling of endoplasmic reticulum membranes and induces motile globules in Nicotiana benthamiana cells.

In infected plant cells, closterovirus replicative polyproteins 1a and 1ab drive membrane remodeling and formation of multivesicular replication platforms. Polyprotein 1a contains a variable Central Region (CR) between the methyltransferase and helicase domains. In a previous study, we have found that transient expression of the Beet yellows virus CR-2 segment (aa 1305-1494) in Nicotiana benthamiana induces the formation of ~1µm mobile globules originating from the ER membranes. In the present study, sequence analysis has shown that a part of the CR named the "Zemlya region" (overlapping the CR-2), is conserved in all members of the Closterovirus genus and contains a predicted amphipathic helix (aa 1368-1385). By deletion analysis, the CR-2 region responsible for the induction of 1-µm globules has been mapped to aa 1368-1432. We suggest that the conserved membrane-modifying region of the BYV 1a may be involved in the biogenesis of closterovirus replication platforms.

Extensive surgical history prior to cytoreductive surgery and hyperthermic intraperitoneal chemotherapy is associated with poor survival outcomes in patients with peritoneal mucinous carcinomatosis of appendiceal origin.

BACKGROUND: Patients with PMCA commonly undergo surgery before CRS/HIPEC. We evaluated the role of extensive surgical treatment before CRS/HIPEC in terms of overall survival (OS). METHODS: 105 patients with PMCA who underwent a CRS/HIPEC procedure were identified from a prospective database. Patients were divided into two groups based on Prior Surgery Score (PSS): PSS ≤ 1 limited surgery group (LSG), PSS >1 extensive surgery group (ESG). Survival of lymph node (LN) negative and positive patients was analyzed separately. RESULTS: 40 patients were in LSG and 65 in ESG. Mean time from diagnosis to CRS/HIPEC was 6 and 17 months for LSG and ESG, respectively (p = 0.004). Groups were well balanced in peritoneal cancer index, complete cytoreduction rate, and LN status. One, 3, and 5-year OS among LN negative patients was 95, 83, and 75% for the LSG (n = 22) group and 87, 55, and 32% for the ESG (n = 35), group respectively (p = 0.026). One, 3, and 5-year OS among LN positive patients was 69, 50, and 17% for the LSG (n = 18) group and 80, 21, and 14% for the ESG (n = 30), group respectively (p = 0.613). For all patients 1, 3, and 5-year OS was 84, 65, and 54% for the LSG (n = 40) group and 86, 43, and 26% for the ESG (n = 65) group, respectively (p = 0.029). CONCLUSION: Extensive surgical treatment before CRS/HIPEC is associated with delay of CRS/HIPEC and poorer OS overall, especially among LN negative patients. We recommend early referral of PMCA patients to a peritoneal surface malignancy center.

Systemic chemotherapy (SC) before cytoreductive surgery and hyperthermic intraperitoneal chemotherapy (CRS/HIPEC) in patients with peritoneal mucinous carcinomatosis of appendiceal origin (PMCA).

BACKGROUND: The role of SC before CRS/HIPEC for patients with PMCA is unclear. This study explores the effect of SC prior to CRS/HIPEC on overall survival (OS) in patients with PMCA. METHODS: 72 patients with recently diagnosed PMCA who underwent CRS/HIPEC were identified from a prospective database. Thirty patients had SC before CRS/HIPEC (Group 1) and 42 did not (Group 2). Patients who were referred to our center after multiple lines of SC were excluded from this analysis. OS was estimated. RESULTS: Median follow-up was 3.2 years. Groups were similar regarding lymph node positivity, postoperative SC and rate of complete cytoreduction. Twenty-four (80%) patients in Group 1 and 21 (50%) in Group 2 had high grade histology (HG) (p = 0.01). OS from CRS/HIPEC at 1, 2, and 3 years was 93, 68, 51% in Group 1 and 82, 64, 60% in Group 2, respectively (p = 0.74). Among HG patients 3-year survival was 36% in the SC group vs. 35% in the group without SC (p = 0.67). The 3-year OS for patients with low grade (LG) tumors was 100% in the SC group vs. 79% in the group with no prior SC (p = 0.26). Among patients with signet ring cell (SRC) histology, 1, 2 and 3-year survival was 94, 67 and 22% in the SC group vs. 43, 14, 14% in the group with no SC, respectively (p = 0.028). There were only 6 patients with LG PMCA who received prior SC. CONCLUSIONS: Preoperative SC could improve the prognosis of patients with high-grade PMCA with SRC histology.

[Ecopsychological investigations in 520-day isolation].

Assessment of the importance of higher plants for emotions and relations within the group of test-subjects was performed in the long-duration chamber experiment MARS-500 designed as an imitation of an exploration mission. It was shown that the degree of involvement with the plants dictated the magnitude and character of the ecopsychological effect. Subjects with high psychological stability and dominating introversion were more inclined to interact with plants. Remoteness of the greenhouse from recreation areas weakened the ecopsychological effect and denied the crew the opportunity to interact with plants. At the same time, the presence of the plant growth unit with decorative plants in the lounge had certainly a positive psychological effect. The crew gave preference to big bright flowers that did not demand much care. This observation should be taken into consideration by designers of psychological recreation environments. Choice of plants to be grown as a dietary supplement for the crew in isolation and confinement should be made with allowance for cultural preferences of crew members.

The role of VEGFR-2 expression in outcomes and survival of patients with peritoneal carcinomatosis from appendiceal cancer.

BACKGROUND: Vascular endothelial growth factor (VEGF) is a prognostic factor and target treatment for metastatic colorectal and ovarian cancer. Cytoreductive surgery (CRS) and hyperthermic intraperitoneal chemotherapy (HIPEC) has improved survival on peritoneal carcinomatosis (PC) from appendiceal cancer. We hypothesize that tumoral high expression of VEGF receptor 2 (VEGFR-2) is a negative prognostic factor for survival in patients with PC from appendiceal cancer. METHODS: A retrospective study of a prospective database revealed 89 patients with PC from appendiceal cancer who underwent 127 CRS/HIPECs. Surgical specimens from 59 patients were tested to identify high vs. low VEGFR-2 expression. Patient outcomes and survival were analyzed. RESULTS: There were 26 males and 33 females. Mean age was 51 years. Forty-seven VEGFR-2 high expressers and 15 low expressers were identified. Mean follow-up of high and low expressers was 25.1 and 26.6 months, respectively (p = 0.806). At follow-up, 33 (70%) high expressers were alive and 14 (30%) deceased, while 11 (92%) low expressers were alive and 1 (8%) deceased. Recurrence, use of bevacizumab, CC score, PCI, and LN status showed no differences between high and low expressers. OS for high expressers was 90.5%, 59.8%, and 47.1% at 1-, 3-, and 5-years, respectively, while OS for low expressers remained stable at 91.7% at 1-, 3-, and 5-years (p = 0.133). CONCLUSION: There is a trend towards better outcomes and survival in patients with PC from appendiceal cancer who have low expression of VEGFR-2 compared to high expression. More studies are encouraged to confirm this trend.

[Association between the person-unique individual behavior styles and the quality and reliability of operator's professional performance].

Among the prioritized objectives of maintaining psychophysiological safety in extended space missions is the buildup of crew professional reliability. Reliability of operator's performance hinges as on skills level, so personal working style. The purpose of the investigation was to penetrate into the patterns of association between a person-unique individual behavior style and quality and reliability of simulated professional operator's jobs. The investigation was part of the MARS-500 experiment (RF SSC - IBMP RAS, 2010-2011) and employed 2 procedures: the well-established manual vehicle control (Pilot-1) and a new one tasking with planetary surface exploration (VIRTU). Observation of individual work styles resulted in identification of 2 fundamentally different approaches to handling the operator's duties--reliable and innovative. It is noteworthy that a separate participant adhered to one and the same style when coping with either procedure. Also, personality traits predisposing to a concrete behavior were found, which confirms the hypothesis of deep personal nature of operator's work style that reveals itself consistently and isomorphically no matter the type of important job.

[Characteristics of the signal lag effect on crew--control center communications in the 520-day simulation experiment].

The 520-day experimental simulation of an exploration mission provided an opportunity to apply content analysis for studying the patterns of crew--Control center (CC) communication impeded by lag times. The period of high autonomy was featured by drastic reduction of the number of crew questions and requests which was judged as a marker of adaptation to the simulated space mission environment. The "key" events in the experiment changed the content of crew messages radically attesting to misperception of time, emotional involvement, want of CC feedback and draining out negative emotions. After the period of high autonomy with full loss of communication with controllers the traffic of crew messages onto the outside was noted to become very light which could also point to temporal changes in the communication style developed in the conditions of isolation and autonomous existence.

Repeated cytoreductive surgery and hyperthermic intraperitoneal chemotherapy in peritoneal carcinomatosis from appendiceal cancer: analysis of survival outcomes.

BACKGROUND: Cytoreductive surgery (CRS)/hyperthermic intraperitoneal chemotherapy (HIPEC) is the procedure of choice in patients with peritoneal dissemination from appendiceal cancer. Although recurrence rates are 26%-44% after first CRS/HIPEC, the role of repeated CRS/HIPEC has not been well defined. We hypothesize that patients undergoing multiple CRS/HIPEC's have meaningful long term survival. METHODS: A retrospective study of a prospective database of 294 patients with peritoneal carcinomatosis (PC) was conducted, of these 162 had PC of appendiceal origin. Twenty-six of these patients underwent 56 CRS/HIPEC. Survival and outcomes was analyzed. RESULTS: The percentage of patients with pre-surgical PCI scores ≥ 20 for the first, second, and third CRS/HIPEC was 65, 65, and 25%, respectively. Complete cytoreduction (CC 0-1) at first, second, and, third surgeries was 96, 65 and 75%, respectively. The mean operating time was 10.1 h. There was no 30-day peri-operative mortality. Following the first, second, and third CRS/HIPEC 27, 42, and 50% experienced grade III complications, respectively. Mean follow up was 51, 28, and 16 months from the first, second, and third CRS/HIPEC, respectively. Overall survival rate for the first CRS/HIPEC was 100, 83, 54, and 46% at years 1, 3, 5 and 10, respectively; from the second CRS/HIPEC 91, 53, and 34% at 1, 3, and 5 years, respectively; and from the third CRS/HIPEC was 75% at one year. CONCLUSION: Repeat CRS/HIPEC can lead to meaningful long term survival rates in patients with appendiceal peritoneal carcinomatosis with morbidity and mortality similar to those of the initial CRS/HIPEC.

[Psychophysiological correlates of individual styles of professional operator's work].

One of the prioritized objectives of psychophysiological support for long-duration space missions is to strengthen professional reliability of cosmonauts. Operator's reliability is dependent as on skillfulness, so individual work style. PILOT-1 and VIRTU methods were used to study individual psychological characteristics and operator's strategy during project MARS500. The established two individual work styles (IWSs), i.e. control and search, reflect different types of reacting to stress. Combined analysis of these data and results of the "Mirror coordinograph" test showed invariance of IWSs no matter whether a task is complicated or simple. Operators demonstrating the "control" strategy are characterized by high initial mobilization readiness. Their reliability demands great physiological resources (high "cost"). Initial mobilization and physiological "cost" are lower in operators demonstrating the "search" work style. Dependent on the level of mobilization and motivation, their efficacy, in terms of quality and reliability specifically, is unstable in nature.