Immunocompetent Mice As a Model for Preclinical Studies of mRNA Vaccine Immunogenicity.

Conducting preclinical studies of mRNA vaccines is complicated by the lack of relevant animal models of the human immune system. Immunocompetent mice are widely used in biomedical research. However, critical differences in the genetics and immune system of mice and humans prevent the study of unique human immune responses in mice. Within the framework of this work, the possibility of modeling the cytotoxic T-cell response to mRNA vaccines encoding the S-protein of the SARS-CoV-2 virus was investigated. High-affinity peptides from S-protein were analyzed for the most frequent allelic variants of human MHC-I, two immunocompetent mouse lines (C57BL/6, BALB/c) and an outbred mouse model of IRC. The results of computer modeling have shown that mouse models can be used in preclinical studies of mRNA vaccines against SARS-CoV-2. Mouse MHC-I is able to present virus peptides that are highly affine for human MHC-I. Moreover, the immunogenicity of some of them has already been confirmed by examining blood samples from patients who have had COVID-19.

Intracellular Gemcitabine Monophosphate Levels Predict Chemotherapy Efficacy in Gemcitabine-Treated Patients with Bladder Cancer.

Gemcitabine monophosphate (dFdCMP), one of the intracellular forms of phosphorylated gemcitabine, determines its antitumor activity. A pharmaco-molecular model for determining relative gemcitabine monophosphate level based on the assessment of the activity of ENT1 and ENT2 channels as well as dCK and CDA enzymes in tumor tissue was developed. Relative gemcitabine monophosphate level is a more relevant predictive factor of gemcitabine resistance of bladder cancer as compared with the expression of individual markers related to dFdCMP formation.

Experimental Study of a Radiopharmaceutical Agent Based on Modified Fatty Acid Labeled with Technetium-99m.

Using rat model of coronary occlusion, we studied pharmacokinetics and the efficiency of a new radiopharmaceutical agent 99mTc-PDA-DTPA intended for diagnostics of changes in myocardial metabolism and its analogue 123I-PMPDA. 99mTc-PDA-DTPA was eliminated mostly by the kidneys and maximal concentration in the heart was attained within 60 min after intravenous injection; no accumulation in the area of myocardial infarction was observed. The studied substance was inferior to its analogue 123I-PMPDA by the quality of scintigraphic visualization of the heart.

[(INDUCED MALARIA)].

Induced malaria continues to be one of the most pressing public health problems in malaria-endemic countries. Ma- laria parasites remain viable in stored blood at a temperature of 2-6°C for 3 weeks. The paper presents current problems associated with transfusion-induced malaria. In the USSR and then in the Russian Federation, sporadic cases of induced malaria (Plasniodium vivax, P.malariae, rarely P.falciparum) were notified (230 cases in 1958 to 1990 and only 5 in 1991 to 2016). Current (immunological and molecular) methods for the laboratory diagnosis of malaria do not provide a 100% detection rMfteor its pathogens; therefore, it is necessary to search for highly efficient, rapid, and low-cost diagnostic methods to ensure the biological safety of donation.

[(EXPERIENCE WITH GEOGRAPHIC INFORRMATION SYSTEMS USED IN EPIDEMIOLOGICAL STUDIES (AS AN EXAMPLE OF MALARIA AND DIROFILARIASIS)].

The effective application of GIS against parasitic diseases requires the patterns of spread of parasitic diseases in certain natural, climatic, and socioeconomic conditions of different regions where there are epidemically effective carriers. The epidemiological parameters defining a high risk of an epidemiological process after P.vivax importation into Russia were calculated using the developed HealthMapper controlled module with a database. Analysis of the average long-term air temperatures in the administrative territories of Russia over 78 years, namely July, the warmest month of the year when there are the largest numbers of all types of mosquito vectors and high levels of microfilaria in the peripheral blood of in- fected dogs (definitive hosts), has shown that the northern border of the maximum possible area of dirofilariasis in Russia is most fully described by the +14C July isotherm.

Riboflavin photoactivation by upconversion nanoparticles for cancer treatment.

Riboflavin (Rf) is a vitamin and endogenous photosensitizer capable to generate reactive oxygen species (ROS) under UV-blue irradiation and kill cancer cells, which are characterized by the enhanced uptake of Rf. We confirmed its phototoxicity on human breast adenocarcinoma cells SK-BR-3 preincubated with 30-µM Rf and irradiated with ultraviolet light, and proved that such Rf concentrations (60 µM) are attainable in vivo in tumour site by systemic intravascular injection. In order to extend the Rf photosensitization depth in cancer tissue to 6 mm in depth, we purpose-designed core/shell upconversion nanoparticles (UCNPs, NaYF4:Yb3+:Tm3+/NaYF4) capable to convert 2% of the deeply-penetrating excitation at 975 nm to ultraviolet-blue power. This power was expended to photosensitise Rf and kill SK-BR-3 cells preincubated with UCNPs and Rf, where the UCNP-Rf energy transfer was photon-mediated with ~14% Förster process contribution. SK-BR-3 xenograft regression in mice was observed for 50 days, following the Rf-UCNPs peritumoural injection and near-infrared light photodynamic treatment of the lesions.

ON THE EPIDEMIOLOGY OF COMPLICATED VIVAX MALRIA.

A larger number of publications on cases of. complicated vivax malaria in the world literature can be. explained by a set of diffeetit faciors'. There has been unification of the definition of complicated tertian malaria allowing ihe clinical nianagement of a patient and the reporting of complicated cases of P.vivax to be improved. The, epidemiological characteristics of complicated vivax malaria are determined by a diversity of geographical, races, variants, gentyies and phenotypes of tertian malaia due to intensive internal and external migration. Chloroquine resistance and primaquine refractoriness contribute to increased local malaria transmission due to a rise in cases of reinfection during-epidemics of vivax malaria. From an epidemiological point of view, the increasing number of complicated vivax malaria cases indicates the untimely detection and treatment of malaria patients, the low level of medical personnel training and a negative attitude of the local population towards the recommendations of national malaria programs for prevention measures. Unsolved problems can substantially retard the time to achieve malaria elimination.

[CLINICAL CASE OF DIROFILARIASIS.]

The paper describes a case of the rare localization of a pregnant Dirofilaria female in a man's genitalia. Patient T born in 1973 was infected in the Krasnoyarsk Territory presumably in the summer of 2012. A migratory mass appeared under the skin of the chest at the beginning of 2016. A dense mass was located beneath the skin in the penis by early April. Histological specimens from the removed tumor, entered the specialized Clinical Diagnostic Laboratory for parasitic diseases. The specimens exhibited transverse and oblique sections of Dirofilaria spp. The body cavity contained nematodes - multiple microfilariae. However, routine studies and PCR could not reveal the microfilariae in the blood.

Antiangiogenic Activity of Alofanib, an Allosteric Inhibitor of Fibroblast Growth Factor Receptor 2.

Alofanib is a potential allosteric inhibitor of FGFR2 used in oncology. The inhibitor blocks the extracellular part of the receptor and prevents its binding with the ligand. Alofanib suppressed proliferation of endothelial cells, their migration activity, and ability to form vessellike structures in vitro and significantly decreased the number of microvessels in Matrigel implant and in ovarian cancer (SKOV-3) xenograft in vivo. The results indicate that Alofanib can inhibit angiogenesis.

[EXPECTED CONSEQUENCES OF TARGETED HUMAN BEHAVIOR MODULATION UPON EXPOSURE TO TOXOPLASMA GONDII].

The paper shows that there are more seropositive people in a group of those who have experienced road accidents (RA) and who are responsible for the latter than in a control group. The investigations have indicated that chronic toxoplasmosis may be regarded as a potential increased risk factor of RA.

[The current status of the resistance of malaria pathogens to antimalarials].

The review presents the results of trials of the clinical efficacy of a test antimalarial drug for each malarial parasite species, which were published in 2000-2013 and supplemented by the data of in vitro studies or investigations using the molecular markers of resistance. There are data on the resistance of each medicament since many of the drugs are used in combination with artermisinin derivatives.

[Epidemiological alertness as a component of the malaria surveillance system].

Malaria vigilance is a continuation of the functions of the malaria surveillance system in the countries that have achieved malaria elimination on their territories. The main function of the malaria vigilance during the post-elimination period is a continuous monitoring and evaluation and implementation of necessary prevention measures against the possible re-establishment of local malaria transmission due to the importation of malaria from its endemic countries by residents and/or migrants. Various epidemiological aspects of malaria vigilance, such as standard identification of a malaria case during the post-elimination period; notification procedures and reporting systems; training of personnel; free access to malaria diagnosis and treatment; mandatory laboratory confirmation of malaria diagnosis; and prompt efficacious treatment, are discussed. Other aspects include epidemiological classification of cases, seasonal pattern of imported cases, its territorial confinement, time-lag between the detection of a case, its diagnosis and treatment initiation, evaluation of the efficacy of individual chemoprophylaxis and treatment.

[The tasks to maintain the achieved results of malaria elimination in the post-elimination period].

The experience of the countries who have achieved malaria elimination on their territories suggests that the main tasks of a malaria program in the post-elimination period are as follows: to maintain healthcare workers' alertness for possible malaria importation from outside and for a risk of its renewed transmission in the malaria-free area; to preserve the medical staff and to improve its professional skills in malaria diagnosis, treatment, and prevention in both the public and private sectors; to maintain malaria alertness among the population, among higher-risk population groups in particular; to elaborate and implement an effective program for collaboration with neighboring malaria-endemic countries, in the boundary regions in particular. The national governments' determination to continue long-term financial support of the prevention program of malaria in its elimination period is the guarantee of successful solution of the tasks set.

[Recombinant fragment of pigment epithelium-derived factor (44-77) prevents pathological corneal neovascularization].

Pigment epithelium-derived factor (PEDF), a 50 kDa secreted glycoprotein, is among the most potent endogenous inhibitors of angiogenesis. PEDF-derived fragment (44-77) possesses antiangiogenic properties of the full-sized protein and is a potential drug candidate for the treatment of ocular neovascular diseases. In this study we propose an efficient scalable biotechnological method for the production of PEDF (44-77) as part of a fusion protein with SspDnaB intein. The fusion protein was obtained in bacterial E. coli cells in the form of inclusion bodies, solubilized and subjected to autocatalytic cleavage with the release of PEDF (44-77) (yield, 77%). The target peptide was separated from the intein using tangential ultrafiltration. The final purification of PEDF (44-77) was performed by reversed-phase HPLC. The yield of the target peptide (purity, 99%) was 65 mg per 1 liter of culture. Antiangiogenic activity of the obtained peptide was studied in vitro using murine endothelial cells SVEC-4-10. PEDF (44-77) suppressed proliferation of endothelial cells by 53% and inhibited endothelial cell tube formation at the concentration of 1 nM. The ability of the recombinant PEDF (44-77) to block initial stages of angiogenesis was demonstrated using the model of rabbit corneal neovascularization.

[pAkt expression in cervical intraepithelial neoplasia (CIN) and in microinvasive cervical cancer].

The study objective was an immunohistochemical evaluation of pAkt expression in 81 CIN and microinvasive cervical cancer tissue samples and 10 samples of relatively "normal" cervical epithelium of HPV-infected women. PAkt expression showed significant up-regulation in CIN2, CIN3 and microinvasive cancer in compare to CIN1 and "normal" epithelium. The rate of pAkt- positive cells increased progressively by cervical neoplasia grade advancement reaching 7 +/- 5% in CIN2, 15 +/- 13% in CIN3 and 17 +/- 15% in microinvasive cancer. The rate of pAkt-positive cases in general was 1,7-fold higher in CIN3 (41%) than in CIN2 (24%). pAkt expression in conjunction with other markers may be used in immunohistochemical studies for individual CIN outcome prognosis and prospectively in immunocytochemical tests for CIN grade diagnostics improvement before using invasive methods. To elaborate multicomponent system of markers with their indexation there is a need for further investigations with greater number of cases.

[Urgent tasks of malaria elimination programs].

The state-of-the-art of malaria elimination programs worldwide and the advances made in the past decade were analyzed and assessed. The factors contributing to or reducing the achievement of the final objective of malaria elimination--to interrupt the local transmission of the infection and to steadily maintain the achieved successes are given. Technical problems are defined at the final stages of a malaria elimination program. The problems include difficulties in identifying patients and asymptomatic parasite carriers due to the low level of malaria transmission and the absence of highly effective methods for detecting malaria parasites with low parasitemia, as well as mixed malaria infections requiring the use of different treatment regimens and antimalarial drugs. Large-scale uncontrolled population migration is noted to be of importance in spreading the infection in a malaria-free area. The solution of urgent tasks is to improve the existing methods and develop new ones for the detection and treatment of the infection and a package of antimalarial measures.

[Ki-67 expression, thymidine phosphorylase and PTEN in intraepithelial cervical carcinoma].

Expression of Ki-67, thymidine phosphorylase (TP) and PTEN were assessed in various grades of cervical intraepithelial neoplasia (CIN) in order to evaluate their potentials of predicting the gravity of possible damage to the epithelium as well as pro- or regression of CIN. Ki-67 and TP levels were shown to correlate directly with CIN grade. It was suggested that a small number of cases of Ki-67 and TP expression absence (15%), exclusively in CIN3 samples, be due to imminent progression to invasive cancer. Both separately and in combination, Ki-67 and TP expression indices should be regarded as having a potential as markers for cervical carcinoma diagnosis, grade and clinical course.

[Use of an enzyme immunoassay test system with cystic Echinococcus antigen to diagnose echinococcosis alveolaris (multilocularis) (alveococcosis)].

Twenty-three serum samples from 12 patients with Echinococcus alveolaris collected 1 to 12 years after surgery and 1--13 courses of specific therapy were used to evaluate the diagnostic effectiveness of an enzyme immunoassay (EIA) test system with cystic Echinococcus antigen. The findings enable the EIA test system with cystic Echinococcus antigen to be recommended for the diagnosis of Echinococcosis alveolaris and for the monitoring of its treatment results.

[Prospects for malaria elimination in Azerbaijan].

Epidemiological analysis of the malaria in the Republic of Azerbaijan has revealed that: 1. In the past year, malaria problem has considerably improved in reducing morbidity and the number of active foci of malaria in the republic. 2. All active foci of malaria have been in its endemic area. 3. Despite the presence of favorable climatic preconditions for malaria in a large part of the republic, socioeconomic preconditions are considerably decreased, causing the malariogenic potential to substantially reduce. 4. All sets a favorable stage for possible interruption of local malaria transmission on the whole territory of the republic provided that financial support for the national malaria elimination program will be increased from the country's government and other sources in conjunction with the implementation of revised malaria control strategy and with the use of current methods for the detection, diagnosis, treatment, and prevention of malaria.

[Use of basidiomycetes in industrial waste processing and utilization technologies: fundamental and applied aspects (review)].

This review provides an analysis of recent data on the mechanisms of degradation of lignocellulosic materials and xenobiotics by basidiomycetes. Special attention is given to the analysis of the current state of research of ligninolytic enzymes and their involvement in the degradation ofxenobiotics. Data on the practical use of basidiomycetes for bioconversion of industrial wastes are systematized. The most promising areas of bioconversion technologies are considered, such as contaminated water purification (including wastewater), cleanup of soils contaminated with heavy metals and xenobiotics, and degradation of difficult-to-degrade substrates (lignin and lignocellulose wastes, low-energy coal, and synthetic polymers).