Immunogenicity of the DNA/Protein Combined Vaccine against COVID-19.

During the COVID-19 pandemic, the development of prophylactic vaccines, including those based on new platforms, became highly relevant. One such platform is the creation of vaccines combining DNA and protein components in one construct. For the creation of DNA vaccine, we chose the full-length spike protein (S) of the SARS-CoV-2 virus and used the recombinant receptor-binding domain (RBD) of the S protein produced in CHO-K1 cells as a protein component. The immunogenicity of the developed combined vaccine and its individual components was compared and the contribution of each component to the induction of the immune response was analyzed. The combined DNA/protein vaccine possesses the advantages of both underlying approaches and is capable of inducing both humoral (similar to subunit vaccines) and cellular (similar to DNA vaccines) immunity.

Immunogenic Properties of the DNA Construct Encoding the Receptor-Binding Domain of the SARS-CoV-2 Spike Protein.

The development of preventive vaccines became the first order task in the COVID-19 pandemic caused by SARS-CoV-2. This paper reports the construction of the pVAX-RBD plasmid containing the Receptor-Binding Domain (RBD) of the S protein and a unique signal sequence 176 which promotes target protein secretion into the extracellular space thereby increasing the efficiency of humoral immune response activation. A polyglucine-spermidine conjugate (PGS) was used to deliver pVAX-RBD into the cells. The comparative immunogenicity study of the naked pVAX-RBD and pVAX-RBD enclosed in the PGS envelope showed that the latter was more efficient in inducing an immune response in the immunized mice. In particular, RBD-specific antibody titers were shown in ELISA to be no higher than 1 : 1000 in the animals from the pVAX-RBD group and 1 : 42 000, in the pVAX-RBD-PGS group. The pVAX-RBD‒PGS construct effectively induced cellular immune response. Using ELISpot, it has been demonstrated that splenocytes obtained from the immunized animals effectively produced INF-γ in response to stimulation with the S protein-derived peptide pool. The results suggest that the polyglucine-spermidine conjugate-enveloped pVAX-RBD construct may be considered as a promising DNA vaccine against COVID-19.

[Immunogenic Properties of the DNA Construct Encoding the Receptor-Binding Domain of the SARS-CoV-2 Spike Protein].

The development of preventive vaccines became the first order task in the COVID-19 pandemic caused by SARS-CoV-2. This paper reports the construction of the pVAX-RBD plasmid containing the Receptor-Binding Domain (RBD) of the S protein and a unique signal sequence 176 which promotes target protein secretion into the extracellular space thereby increasing the efficiency of humoral immune response activation. A polyglucine-spermidine conjugate (PGS) was used to deliver pVAX-RBD into the cells. The comparative immunogenicity study of the naked pVAX-RBD and pVAX-RBD enclosed in the PGS envelope showed that the latter was more efficient in inducing an immune response in the immunized mice. In particular, RBD-specific antibody titers were shown in ELISA to be no higher than 1 : 1000 in the animals from the pVAX-RBD group and 1 : 42000, in the pVAX-RBD-PGS group. The pVAX-RBD-PGS construct effectively induced cellular immune response. Using ELISpot, it has been demonstrated that splenocytes obtained from the immunized animals effectively produced INF-y in response to stimulation with the S protein-derived peptide pool. The results suggest that the polyglucine-spermidine conjugate-enveloped pVAX-RBD construct may be considered as a promising DNA vaccine against COVID-19.

Effect of Detergents on Activity and Magnesium-Dependent Properties of Different Isoforms of Na+,K+-ATPase in the Crude Membrane Fraction of Rat Cerebral Cortex.

We studied the effect of various detergents (Tween-20, Triton X-100, and sodium deoxycholate) on activity and magnesium-dependent properties of Na+,K+-ATPase of the crude membrane fraction of rat cerebral cortex. All studied detergents significantly increased activity of the studied enzyme in a concentration-dependent manner. Sodium deoxycholate provided significantly higher values Na+,K+-ATPase activity (by ≈50%) than Triton X-100 and Tween-20. In the presence of Triton X-100, a changed pattern of the dependence of enzyme activity on the concentration of magnesium ions in the incubation solution was noted. Separate measurement of activities of Na+,K+-ATPase isoforms made it possible to assume that changes in magnesium-dependent properties are due to the predominant effect of Triton X-100 on ouabain-sensitive α2- and α3-isoforms.

mRNA technology as one of the promising platforms for the SARS-CoV-2 vaccine development.

After the genome sequence of SARS-CoV-2 (Severe acute respiratory syndrome-related coronavirus 2) was published and the number of infected people began to increase rapidly, many global companies began to develop a vaccine. Almost all known approaches to vaccine design were applied for this purpose, including inactivated viruses, mRNA and DNA-vaccines, vaccines based on various viral vectors, synthetically generated peptides and recombinant proteins produced in cells of insects and mammals. This review considers one of the promising vaccine platforms based on messenger RNA. Until recent years, mRNA-vaccination was out of practical implementation due to high sensitivity to nuclease degradation and consequent instability of drugs based on mRNA. Latest technological advances significantly mitigated the problems of low immunogenicity, instability, and difficulties in RNA-vaccine delivery. It is worth noting that mRNA-vaccines can efficiently activate both components of the immune system, i. e. T-cell and humoral responses. The essential advantage of mRNAvaccines includes fast, inexpensive, scalable and uniform production providing a large output of desirable products in vitro. Synthesis and purification processes significantly simplify the process technology of mRNA drugs with injectable purity. Thus, mRNA production via in vitro transcription is more advantageous as compared with DNA-vaccines since it is a chemical process without the use of cells. mRNA techniques make it possible to pass all the phases of vaccine development much faster in comparison with the production of vaccines based on inactivated viruses or recombinant proteins. This property is critically important when designing vaccines against viral pathogens as the main problem of disease control includes a time gap between an epidemic and vaccine development. This paper discusses studies on the development of vaccines against coronaviruses including SARS-CoV-2 with special attention to the mRNA technique.

Pattern of extinction of the woolly mammoth in Beringia.

Extinction of the woolly mammoth in Beringia has long been subject to research and speculation. Here we use a new geo-referenced database of radiocarbon-dated evidence to show that mammoths were abundant in the open-habitat of Marine Isotope Stage 3 (∼45-30 ka). During the Last Glacial Maximum (∼25-20 ka), northern populations declined while those in interior Siberia increased. Northern mammoths increased after the glacial maximum, but declined at and after the Younger Dryas (∼12.9-11.5 ka). Remaining continental mammoths, now concentrated in the north, disappeared in the early Holocene with development of extensive peatlands, wet tundra, birch shrubland and coniferous forest. Long sympatry in Siberia suggests that humans may be best seen as a synergistic cofactor in that extirpation. The extinction of island populations occurred at ∼4 ka. Mammoth extinction was not due to a single cause, but followed a long trajectory in concert with changes in climate, habitat and human presence.

[An extracellular pH changes registration by confocal microscopy in higher plant at the excitation potentials generation].

Confocal microscopy technique was applied for registration of apoplast pH changes in the Cucurbita pepo seedling stem during generation of action potential and variation potential. Fluorescent dye FITC-dextran was used for pH-changes registration. Analysis of fluorescence images and fluorescence spectra showed, that FITC-dextran was localized in cell walls. Propagation of action potential and variation potential was accompanied by transient increase of fluorescence intensity, which indicated alkalization of cell walls. This transient alkalization is proposed to be caused by a temporary inactivation of the H(+)-pump in the plasma membrane.

[Psychophysiological aspects of duodenal ulcer in young patients]. / Psikofiziologicheskie aspekty iazvennoi bolezni dvenadtsatiperstnoi kishki u lits molodogo vozrasta.

SMALL and Spilberg-Khanin questionnaires were used to examine psychological state while statistical and spectral analysis of heart rate at rest and orthostasis to evaluate vegetative status in 86 patients aged 15-24 years with duodenal ulcer (DU). Patients with mild DU had the least psychological and vegetative alterations primarily of the neurotic trend with a relative enhancement of the nervous sympathetic component. A severe disease was characterized by the greatest psychovegetative changes--high SMALL scales, anxiety, stress of the vegetative nervous systems at rest and orthostasis. Thus, alterations in psychic and vegetative status may serve criteria in prediction of DU course in young patients.

[Use of organic forms of selenium in nutrition of patients with gastrointestinal diseases]. / O primenenii organicheskoi formy selena v pitanii gastroénterologicheskikh bol'nykh.

Selenium level was determined in sera of 27 patients including 15 patients with chronic gastrointestinal diseases and 12 patients with intestinal malabsorption after some kinds of stomach or small intestine resection. Deficiency of this trace element was found in 5 patients and suboptimal concentration in 4 patients in the first group and in 3 and 5 patients of second group respectively. Supplementation of patients with 45 mg of selenium organic form based on autolysis extract of selenious baker's yeast resulted in increase of selenium level in 23 (85%) patients including all of them with selenium deficiency or suboptimal level. It was concluded about high bioavailability of selenium in supplement studied. Perturbed intestinal absorption after surgical resection of stomach or small intestine may be considered as an indication for prophylactic taking of organic selenium form preparation.

[Study of the state of lipid peroxidation system: antioxidant protection during the use of soy oil in patients with ischemic heart disease and hypertension]. / Izuchenie sostoianiia sistemy perekisnogo okisleniia lipidov--antiokislitel'noi zashchity pri ispol'zovanii soevogo masla u bol'nykh ishemicheskoi bolezn'iu serdtsa i gipertonicheskoi bolezn'iu.

Usage of antisclerotic diet with PUFA n-3 from soy oil in patients with IHD and HBP resulted in positive dynamic of clinic manifestation, blood lipids and coagulogramms of the patients. The decrease of intensity of lipid peroxidation was also revealed. The results showed that soy oil may be recommended for preventive and treatment diets.

[The comparative assessment of follow-up for hypertensive patients]. / Sravnitel'naia kharakteristika dispansernogo nabliudeniia za patsientami s arterial'noi gipertenziei.

The quality of outpatient follow-up was compared in hypertensive patients of the primary care system in 1995 vs 1997. Case records have been retrospectively analyzed for 1301 patients and 1335 ones treated in 6 Kemerovo outpatient clinics in 1995 and 1997, respectively. A special sheet was used for recording the data. More investigations of the target organs were made in 1997. An increase was found in the number of tests for total cholesterol, creatinine, blood glucose, blood count, urinalysis, ultrasound scanning of the kidneys and adrenals, ECG. More patients were taking drugs. Adelphan administration decreased by 32%, clophelin by 30%. More frequently were prescribed ACE inhibitors, calcium antagonists, beta-adrenoblockers. Quality of care for hypertensive patients is still low. This is explained by deficient social care, poor medical education of the patients, rejection of active treatment by many patients.

[Effects of mineral contents of a ration on indicators of radiation resistance of the body]. / Vliianie mineral'nogo sostava ratsiona pitaniia na pokazateli radiorezistentnosti organizma.

Relationship between the levels of calcium, potassium, and iron in rations and the indexes of antioxidant system of rats exposed to prolonged 137Cs loading was studied. Calcium deficiency was found to weaken the antioxidant system. Increased intake of potassium and iron provide high level of adaptation of the antioxidant system.

[The antioxidant defense system in hypertension patients on a diet enriched with omega 3 polyunsaturated fatty acids and alpha-tocopherol]. / Issledovanie sistemy antiokislitel'noi zashchity u bol'nykh gipertonicheskoi bolezn'iu pri obogashchenii diety polinenasyshchennymi zhirnymi kislotami omega3 i alpha-tokoferolom.

A state of the anti-oxidation system in blood and the rate of lipid peroxidation were studied in patients with hypertension. High sensitivity of the anti-oxidation system enzymes to alterations in the diet fat composition was detected in the patients, nutrition of whom was evaluated in detail. Excessive doses of alpha-tocopherol, added to the diet enriched with omega 3-polyunsaturated fatty acids normalized lipid peroxidation and stabilized the enzyme activity involved in the anti-oxidation system. The integral parameter, proposed for evaluation of the anti-oxidation protection state and designated as "anti-oxidation index" involved estimation of catalase, superoxide dismutase, glutathione reductase and peroxidase activities as well as of content of malonic dialdehyde and diene conjugates in erythrocytes.

[Optimization of determining human erythrocyte glutathione reductase activity using a semiautomatic analyzer]. / Optimizatsiia opredeleniia aktivnosti glutationreduktazy éritrotsitov cheloveka na poluavtomaticheskom analizatore.

A relatively simple procedure was developed for estimation of the glutathione reductase activity using half-automatic analyzer. The procedure involved: preparation of hemolysates with minimal rate of erythrocyte destruction; substitution of buffers with high ionic strength for 0.05 M Tris-HCl buffer not containing EDTA. Most effective period of preincubation was found; dependence of the reaction rate on FAD was studied. The procedure developed was approved using two types of half-automatic analyzer and the model FP-901 was shown to be the most suitable for estimation of the glutathione reductase holoenzyme activity.

[Effect of anti-arteriosclerosis diet, containing soya protein isolate and omega-3 polyunsaturated fatty acids on the activity of mononuclear and platelet lysosomal hydrolases in patients with hypertension and ischemic heart disease]. / Vliianie protivoateroskleroticheskoi diety, soderzhashchei izoliat soevogo belka i polinenasyshchennye zhirnye kisloty omega-3, na aktivnost lizosomalnykh gidrolaz mononuklearov i trombotsitov bolnykh ishemicheskoi bolezníu serdtsa i gipertonicheskoi bolezníu

In response to antiatherosclerosis dietotherapy containing 20 g of ichthyenic oil, coronary and hypertensive subjects showed lowered serum levels of cholesterol, triglycerides and atherogenic index, elevated HDLP cholesterol and corrected immunochemical shifts. SPI-containing diet resulted in changes of CIC IgM levels only. Shifts in the activity of mononuclear and platelet lysosomal hydrolases which occurred in the above patients due to relevant diets reflect higher sensitivity of this parameter in assessment of the dietotherapy effectiveness.

[The effect of a soy protein isolate and sources of polyunsaturated omega-3 fatty acids in an anti-atherosclerotic diet on the lipid spectrum of blood serum and immunological indicators in patients with ischemic heart disease and hypertension]. / Vliianie izoliata soevogo belka i istochnikov polinenasyshchennykh zhirnykh kislot omega3 v sostave protivoateroskleroticheskoi diety na lipidnyi spektr syvorotki krovi i immunologicheskie pokazateli u bol'nykh ishemicheskoi bolezn'iu serdtsa i gipertonicheskoi bolezn'iu.

Blood serum lipid spectra and immunologic spectra were studied in patients with ischemic heart disease and hypertension maintained on antiatherosclerotic diet containing either soy bean protein isolates or ichthyenic oil. The ichthyenic oil-containing diet decreased distinctly the level of cholesterol, triglycerides and the atherogenicity coefficient simultaneously with an increase of cholesterol content in high density lipoproteins in blood of 37 patients with these diseases. The diet with soy bean protein isolates exhibited less pronounced effects on lipid metabolism patterns in blood serum as compared with the ichthyenic oil-containing diet; however, the former diet contributed to normalization of the immunologic reactions in patients with ischemic heart disease and hypertension.

[Oxygen therapy of gonarthrosis deformans]. / Kislorodoterapiia v lechenii deformiruiushchego gonartroza.

In the article is presented the comparative estimate of the oxygen therapy methods in treatment of the deforming gonarthrosis. The new method of treatment is described, including 4-5 courses of hyperbaric oxygenation therapy at 1-1.5 atms during 50 min and intraarticular introduction of oxygen. 3 groups of patients have been singled out. The proposed method have been applied with 32 patients. It turned out to be more effective and the course of treatment shorter as compared with the known methods: with application of hyperbaric oxygenation without local introduction of oxygen and localized introduction of oxygen in the joint without application of hyperbaric oxygenation.

[Clinico-biochemical evaluation of the effectiveness of diet therapy in gastroenterologic patients]. / Kliniko-biokhimicheskaia otsenka éffektivnosti dietoterapii gastroénterologicheskikh bol'nykh.

Dissimilar immunochemical, protein and vitamin parameters were found in blood of patients with gastroenterological diseases at the initial step of impairments under conditions of routine and special diets. Specific alterations in content of transferrin, haptoglobin, tocopherol as well as in glutathione peroxidase and glucose-6-phosphate dehydrogenase activities showed that compensatory mechanisms were maintained better under conditions of the special diet therapy.

[Experience in treating senile and seborrheic keratomata by cryogenic action with liquid nitrogen]. / Opyt lecheniia senil'nykh i seboreinykh keratom metodom kriogennogo vozdeistviia zhidkim azotom.

A total of 189 patients with senile keratosis and 490 with seborrheic keratomata have been treated with cryotherapy. Relapses have been recorded in 6.1% of patients.