Use of Adjuvant Compositions Based on Squalene Ensures Induction of Neutralizing Antibodies against SARS-CoV-2.

Squalene-based adjuvant compositions that can provide effective induction of specific humoral immune response have been developed. Recombinant receptor-binding domain (RBD) of surface S-protein of SARS-CoV-2 was used to evaluate the properties of the composition. Immunization of mice with the developed squalene-based compositions in combination with RBD allows obtaining high titers of specific antibodies: from 105 to 2×106. The blood sera from immunized mice exhibit neutralizing activity against SARS-CoV-2 Delta variant (B.1.617.2) with a titer up to 1:2000.

The Oldest Swallow (Aves: Passeriformes: Hirundinidae) from the Upper Lower Miocene of Southeastern Siberia.

A new genus and species of fossil swallows, Miochelidon eschata gen. et sp. nov., are described from the Lower Miocene of the Baikal region. This is the only Miocene representative of Hirundinidae and the oldest fossil record of the group. The coracoid of Miochelidon combines generalized and specialized features, suggesting a basal position in the family for the genus. The ulna has a more advanced morphology, like in modern swallows. The find indicates a widespread distribution of basal swallows in the Early Miocene.

Problems of creating antibody phage libraries and their solutions.

Phage display has become an efficient, reliable and popular molecular technique for generating libraries encompassing millions or even billions of clones of divergent peptides or proteins. The method is based on the correspondence between phage genotype and phenotype, which ensures the presentation of recombinant proteins of known amino acid composition on the surface of phage particles. The use of affinity selection allows one to choose variants with affinity for different targets from phage libraries. The implementation of the antibody phage display technique has revolutionized the field of clinical immunology, both for developing tools to diagnose infectious diseases and for producing therapeutic agents. It has also become the basis for efficient and relatively inexpensive methods for studying protein-protein interactions, receptor binding sites, as well as epitope and mimotope identification. The antibody phage display technique involves a number of steps, and the final result depends on their successful implementation. The diversity, whether natural or obtained by combinatorial chemistry, is the basis of any library. The choice of molecular techniques is critical to ensure that this diversity is maintained during the phage library preparation step and during the transformation of E. coli cells. After a helper phage is added to the suspension of transformed E. coli cells, a bacteriophage library is formed, which is a working tool for performing the affinity selection procedure and searching for individual molecules. Despite the apparent simplicity of generating phage antibody libraries, a number of subtleties need to be taken into account. First, there are the features of phage vector preparation. Currently, a large number of phagemid vectors have been developed, and their selection is also of great importance. The key step is preparing competent E. coli cells and the technology of their transformation. The choice of a helper phage and the method used to generate it is also important. This article discusses the key challenges faced by researchers in constructing phage antibody libraries.

The Search for Single-Domain Antibodies Interacting with the Receptor-Binding Domain of SARS-CoV-2 Surface Protein.

We performed a search for nanoantibodies that specifically interact with the receptor-binding domain (RBD) of the SARS-CoV-2 surface protein. The specificity of single-domain antibodies from the blood sera of a llama immunized with RBD of SARS-CoV-2 surface protein S (variant B.1.1.7 (Alpha)) was analyzed by ELISA. Recombinant trimers of the SARS-CoV-2 spike protein were used as antigens. In this work, a set of single-domain antibodies was obtained that specifically bind to the RBD of the SARS-CoV-2 virus.

[Association of the structure of the glucocorticoid receptor and single nucleotide NR3C1 gene polymorphisms with metabolic disorders].

Glucocorticoid therapy is widely used in the treatment of various pathologies. Sensitivity to glucocorticoids  (GC) has a serious impact not only on the effectiveness of their action, but also on the severity of side effects, the formation of risk factors and the development  of cardiovascular diseases (CVD). Variability of sensitivity to GC causes different phenotypes and severity of metabolic disorders underlying  CVD. Among  them, one can distinguish  a decrease in muscle mass and strength, obesity, glucose and lipid metabolism impairment, and others. Glucocorticoids carry out their effects by binding to the glucocorticoid receptor (GR), and therefore this is considered a critical point in their action. This review presents data on the significance of the glucocorticoid  receptor structure, examines the main single nucleotide polymorphisms (SNP) of the NR3C1 gene associated with hypersensitivity  or relative resistance to glucocorticoids  in the context of metabolic disorders and the development of CVD. The association of the four most studied SNP of the GR gene with metabolic risks is described in detail: BclI (rs41423247), N363S (rs56149945), ER22/23EK (rs6189/rs6190), GR-9ß (rs6198). Their determination can contribute to clarifying the prognosis of both the effectiveness of GC and the development of metabolic disorders, and subsequent early correction of CVD risk factors.

The GWAS-MAP|ovis platform for aggregation and analysis of genome-wide association study results in sheep.

In recent years, the number of genome-wide association studies (GWAS) carried out for various economically important animal traits has been increasing. GWAS discoveries provide summary statistics that can be used both for targeted marker-oriented selection and for studying the genetic control of economically important traits of farm animals. In contrast to research in human genetics, GWAS on farm animals often does not meet generally accepted standards (availability of information about effect and reference alleles, the size and direction of the effect, etc.). This greatly complicates the use of GWAS results for breeding needs. Within the framework of human genetics, there are several technological solutions for researching the harmonized results of GWAS, including one of the largest, the GWAS-MAP platform. For other types of living organisms, including economically important agricultural animals, there are no similar solutions. To our knowledge, no similar solution has been proposed to date for any of the species of economically important animals. As part of this work, we focused on creating a platform similar to GWAS-MAP for working with the results of GWAS of sheep, since sheep breeding is one of the most important branches of agriculture. By analogy with the GWAS-MAP platform for storing, unifying and analyzing human GWAS, we have created the GWAS-MAP|ovis platform. The platform currently contains information on more than 34 million associations between genomic sequence variants and traits of meat production in sheep. The platform can also be used to conduct colocalization analysis, a method that allows one to determine whether the association of a particular locus with two different traits is the result of pleiotropy or whether these traits are associated with different variants that are in linkage disequilibrium. This platform will be useful for breeders to select promising markers for breeding, as well as to obtain information for the introduction of genomic breeding and for scientists to replicate the results obtained.

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.

Negative heterosis for meiotic recombination rate in spermatocytes of the domestic chicken Gallus gallus.

Benef its and costs of meiotic recombination are a matter of discussion. Because recombination breaks allele combinations already tested by natural selection and generates new ones of unpredictable f itness, a high recombination rate is generally benef icial for the populations living in a f luctuating or a rapidly changing environment and costly in a stable environment. Besides genetic benef its and costs, there are cytological effects of recombination, both positive and negative. Recombination is necessary for chromosome synapsis and segregation. However, it involves a massive generation of double-strand DNA breaks, erroneous repair of which may lead to germ cell death or various mutations and chromosome rearrangements. Thus, the benef its of recombination (generation of new allele combinations) would prevail over its costs (occurrence of deleterious mutations) as long as the population remains suff iciently heterogeneous. Using immunolocalization of MLH1, a mismatch repair protein, at the synaptonemal complexes, we examined the number and distribution of recombination nodules in spermatocytes of two chicken breeds with high (Pervomai) and low (Russian Crested) recombination rates and their F1 hybrids and backcrosses. We detected negative heterosis for recombination rate in the F1 hybrids. Backcrosses to the Pervomai breed were rather homogenous and showed an intermediate recombination rate. The differences in overall recombination rate between the breeds, hybrids and backcrosses were mainly determined by the differences in the crossing over number in the seven largest macrochromosomes. The decrease in recombination rate in F1 is probably determined by diff iculties in homology matching between the DNA sequences of genetically divergent breeds. The suppression of recombination in the hybrids may impede gene f low between parapatric populations and therefore accelerate their genetic divergence.

[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 Xymedon and Mexidol in Combination with Antineoplastic Drugs on Spermatogenesis Indicators and Functional State of Spermatozoa in Rats with Walker-256 Carcinoma.

We compared the effect of Xymedon (100 mg/kg), Mexidol (50 mg/kg), and their combination on spermatogenesis indicators and functional state of spermatozoa in rats with Walker-256 carcinoma treated with doxorubicin (4 mg/kg) and cyclophosphamide (45 mg/kg) (once intraperitoneally on day 11 after tumor cells transplantation). Xymedon and Mexidol were injected intramuscularly for 10 days starting from day 11 of the experiment. The studied parameters were evaluated on experimental days 14 and 21. We have established that gonadoprotective effect of Xymedon developed gradually and persisted longer than that of Mexidol. It manifested in an increase in the number of epithelial spermatogenesis cells (spermatogonia by 3.2 times, early spermatids by 2.2 times, late spermatids by 2.9 times, and Leydig cells by 4 times) in the testes and also the proportion of viable progressively and non-progressively motile epididymal spermatozoa (by 2 times). The combination of Xymedon and Mexidol stimulated spermatogenesis (with restoration of the initial level of spermatocytes, an increase in the number of early spermatids by 65.5 and 99% in comparison with Xymedon alone and Mexidol alone, respectively) and increased the number of viable epididymal spermatozoa more effectively than Xymedon and Mexidol alone by 54 and 60%, respectively.

Effect of the Entomopathogenic Fungus Beauveria bassiana on the Development of Faba Bean (Vicia faba) Diseases in the Field Conditions.

Several ascomycetous entomopathogenic fungi, including species in the genera Beauveria, are plant symbionts/endophytes and are termed as endophytic insect-pathogenic fungi. It was shown that the fungus Beauveria bassiana (BBK-1 strain) successfully colonized Vicia faba bean plants in laboratory and field conditions of Western Siberia. The B. bassiana reisolate passed through the plants had significantly higher antagonistic activity against phytopathogens in comparison with the primary stem of entomopahogenic fungi. Pre-sowing faba bean seeds treatment reduced the level of infection of the seed material with phytopathogens, significantly decrease the development and prevalence of root rot disease. A decrease in the disease development index (chocolate spot, powdery mildew, fusariosis and other spots diseases) was found as a result of the use of B. bassiana. The effectiveness and prolonged action of B. bassiana on plants opens up new opportunities both in the creation of biological products and in molecular-genetic research and selection of certain pairs of plants and fungi based on the principle of the greatest synergy.

Prototype of a DNA Vaccine against Marburg Virus.

The preparation and study of the biological properties of the pVAKS-GPVM DNA immunogen containing a gene encoding Marburgvirus glycoprotein are described. The specificity of blood serum antibodies of guinea pigs immunized with DNA immunogen was analyzed by ELISA. Inactivated viral preparation, recombinant glycoprotein (GP) obtained in the prokaryotic system and virus-like particles based on the recombinant vesicular stomatitis virus exhibiting Marburgvirus GP were used as the antigens. The neutralizing activity of antibodies of immunized animals was tested in vitro using a pseudovirus system. It was demonstrated that the developed immunogen administered to guinea pigs induced the production of specific antibodies that neutralize virus-like particles and Marburgvirus in cultured Vero cells.

[Nd:YAG laser goniopuncture as a mandatory adjuvant procedure after non-penetrating deep sclerectomy (long-term observation results)]. / Nd:YAG-lazernaia goniopunktura kak obiazatel'naia ad''iuvantnaia protsedura posle nepronikaiushchei glubokoi skleréktomii (rezul'taty dolgosrochnogo nabliudeniia).

Today, Nd:YAG laser goniopuncture (LGP) is considered a mandatory non-penetrating deep sclerectomy adjuvant procedure. However, its indications and timing remain debatable. PURPOSE: To evaluate the effect of Nd:YAG laser goniopuncture on the long-term hypotensive effectiveness of non-penetrating deep sclerectomy. MATERIAL AND METHODS: The study included 114 patients after non-penetrating deep sclerectomy (NPDS). In the control group (n=58), Nd:YAG laser goniopuncture was performed within 3.4±1.9 (1.5-6.7) months, and in the main group (n=56) - within 1.12±0.08 (0.9-1.5) months after the surgery. Ultrasound biomicroscopy (UBM) was used to evaluate the semiotics of trabecular-Descemet's membrane (TDM), intrascleral canal (ISC) and filtering bleb. The follow-up period was 5 years. RESULTS: According to UBM data, the thickness (0.10±0.009 mm) and density (50±6%) of TDM (p=0.0001) increased before LGP in the control group, the height of ISC decreased to 0.49±0.19 (0.20-0.40) (p=0.03), the height of UBM scan - to 1.49±0.05 (1.41-2.9) (p=0.0001); IOP (P0) was 18.48±4.7 (11.2-22.9) mmHg (p=0.001). In the main group before LGP, TDM thickness was 0.08±0.006 mm, density was 40±5%, and IOP (P0) was 15.7±4.1 (9.1-18.5) mm Hg. Complete hypotensive success was achieved in 83.6% of cases in the control group and 96.2% in the main group in 6 months; 68.07% and 92.59% in 12 months; 41.3% and 75.8% in 24 months; 15.25% and 48.93% in 36; 15% and 34.8% in 60 months after the surgery, respectively (p=0.0001, 95% confidence interval). CONCLUSION: TDM is an additional level of retention of aqueous humor and plays key role in the formation of outflow pathways after NPDS. Performing LGP in the early postoperative period is an effective and safe adjuvant option, which excludes the influence of TDM on the formation of aqueous humor outflow pathways and significantly increases the long-term hypotensive efficacy of non-penetrating deep sclerectomy.

Clinical significance of lipohypertrophy without visual and palpable changes detected by ultrasonography of subcutaneous fat.

AIM: To estimate clinical significance of lipohypertrophy (LH) without visual and palpable changes, detected by ultrasonography of subcutaneous fat. MATERIALS AND METHODS: This study included 140 diabetic patients who received insulin in basal-bolus regimen. Ultrasonography of subcutaneous fat was performed for LH diagnostics in these diabetic patients. Than clinical significance of LH without visual and palpable changes was estimated. HbA1c level, fasting and postprandial glucose, episodes of hypoglycemia, body mass index (BMI) and scheme of insulinotherapy were evaluated at the moment of LH, after 3 and 6 months in all patients. RESULTS: After changing injection sites, good results were demonstrated by measuring glucose and HbA1c level. Thus fasting glucose decreased from 9.03±1.98 mmol/l to 7.11±0.95 mmol/l (p=0.023). Postprandial glucose reduced from 10.27±2.72 mmol/l to 9.34±1.21 mmol/l (p=0.011). HbA1c level reduced from 9.27±1.75% to 7.43±1.02% (p=0.002). Also BMI decreased from 33.75±3.49 kg/m2 to 30.5±2.96 kg/m2 (p=0.018). CONCLUSION: LH without visual and palpable changes could worsen compensation of glycemic control and leads to hypoglycemia and chronic Somogyi rebound. So, LH without visual and palpable is as important and clinically significant as classic LH.

[Correlation of parameters of carbohydrate metabolism and saturated fatty acids in blood serum in elderly people.]

The study involved 106 elderly people of the Arctic aged 61-74 years, 42 of them were residents of the Subarctic region (SR) and 64 residents of the Arctic region (AR). Using gas-liquid chromatography there was determined the serum content of saturated fatty acids (SFA): short-chain (SCFA), medium-chain (MCFA) and long-chain (LCFA); and the levels of carbohydrate metabolism parameters were determined by spectrophotometric methods. Correlation analysis showed that elderly people of SR had no significant interrelationships between the contents of SCFA, MCFA and LCFA, since all the correlation dependences were weak (r=0,2-0,29, p=0,08-0,786), against AR with a lower occurrence of high values of glucose (Glu), lactate (Lac) and low pyruvate (Pyr). In AR, with the highest occurrence of high and low values of GIu, the greatest number of its interrelations was noted with LCFA (palmitic, margarine, stearic, behenic, tricosanoic), then with MCFA (myristic, pentadecanoic) and SCFA (pelargonic), low values of Pyr with MCFA (lauric, myristic, ∑MCFA) and SCFA (decanoic), and a slight decrease in the occurrence of high levels of Lac and Lac/Pyr with LCFA (palmitic, margarine, stearic, tricosanoic), MCFA (lauric, tridecanoic) and SCFA (caproic, capric).

INTERRELATION OF THE GENE 4DKN1A (RS 1801270) POLYMORPHIC STATE AND LEVEL OF DEVELOPMENT OF CARIES IN CHILDREN WITH DOWN SYNDROME.

Study is devoted for rs1801270 polymorphism of CDKN1A gene due to the level of intensity of caries development in children with Down syndrome. The full development of oral cavity organs is ensured through a complex of genes whose protein products control different stages of this difficult process. Changes in gene structure, as well as disturbance of their expression may lead to abnormalities of development of teeth and periodontal tissues. Analysis of the polymorphism of CDKN1A gene Ser31Arg in children living in Kharkiv region has shown that the frequency of genotypes and alleles corresponds to the theoretically expected distribution by Hardy-Weinberg in the core (χ2=0.12) and control (χ2=1.29) groups. There is no significant differences in the frequency of alleles of CDKN1A gene in the group of healthy children and children with Down syndrome (p=0.978). There is an association of heterozygous option C/A of the studied marker Ser31Arg of CDKN1A gene with the level of intensity of caries development (by Leus) in children with Down syndrome under 10 years old.

[Nonalcoholic fatty liver disease: What do we know and what will we have to learn?] / Nealkogol'naia zhirovaia bolezn' pecheni: chto my znaem i chto predstoit uznat'.

The article reviews relevant data on the prevalence, natural history, pathogenesis, diagnosis, and treatment of nonalcoholic fatty liver disease and critically assesses the fixed notion of this disease.

Novel method for facial rejuvenation using Er:YAG laser equipped with a spatially modulated ablation module: An open prospective uncontrolled cohort study.

Facial aging is a complex biological process that affects the skin and superficial musculoaponeurotic system (SMAS). A new technology (RecoSMA) for skin rejuvenation based on acoustic-interference method using Er:YAG laser (2936 nm) equipped with a special module SMA that targets both the dermis and SMAS was evaluated in an open-label prospective cohort study of 100 female patients treated for facial rejuvenation. Measure of clinical improvement included investigator-rated clinical photography using the Modified Fitzpatrick Wrinkle Scale, and ultrasound measurements in the dermis a week, 30 days and six months post treatment. All patients completed the study and no complications were noted. Improvements in skin tone and texture were noted in all participants and significant decrease in wrinkle depth was demonstrated at the six-month follow-up that was confirmed by ultrasound skin measurements. Data presented herein confirm the safety and efficacy of RecoSMA treatment for facial rejuvenation.