SYSTEMIC INFLAMMATORY INDICES IN PATIENTS WITH MALIGNANT GLIOMAS AND EFFECTS OF PLATELET SECRETOME IN VITRO.

BACKGROUND: To date, no significant clinical progress has been achieved in the treatment of brain malignant gliomas (MG), and the active search for non-invasive circulating biomarkers continues. The prognostic significance of the ratio of the main peripheral blood cell populations of patients with MG is evaluated. Considerable attention is paid to the secretome of platelets (Pt) of peripheral blood. AIM: To evaluate the indicators of the peripheral blood cell population ratios in patients with brain MG and to study the influence of the secretome of Pt (SPt) of the peripheral blood of patients with brain MG in cell cultures in vitro. MATERIALS AND METHODS: We studied samples of peripheral blood from patients with glioma CNS WHO grade G2 (n = 5), G3 (n = 12), and G4 (n = 20). The peripheral blood cell counts were analyzed in the preoperative period on an automatic hematology analyzer. The in vitro study of SPt was performed on the U251 human glioblastoma cell line cultured with SPt from MG patients or SPt pre-incubated with anti-TGF-ß1 antibody. Cell cultures were observed for 72 h, and mitotic index (MI) was calculated. RESULTS: In MG patients, the count of peripheral blood leukocytes and neutrophils increased (p < 0.05). The neutrophil-to-lymphocyte ratio (NLR) and systemic immune-inflammation index (SII) increased by 2-3 times compared to control. Nevertheless, correlation analysis did not reveal significant relationships between quantitative indicators of peripheral blood cells and the tumor malignancy degree in MG patients. The MI in U251 cells increased under the influence of SPt from patients with MG (p < 0.021), correlated with the tumor degree of malignancy (r = 0.246, p = 0.014). Pre-incubation of SPt with anti-TGF-ß1 antibody tends to neutralize this promitotic effect. CONCLUSION: In MG patients, the integral indicators of NLR and SII increased but no significant relationship with the degree of tumor malignancy was found. In U251 cells, promitotic effects of SPt of MG patients partially decreased by anti-TGF-ß1 antibody.

[Bacteriophages in complex treatment of septic diseases of the hand]. / Ispol'zovanie lekarstvennykh preparatov bakteriofagov v kompleksnom lechenii gnoinykh zabolevanii kisti.

OBJECTIVE: To evaluate the effectiveness of bacteriophages in complex surgical treatment of septic diseases of the hand. MATERIAL AND METHODS: We analyzed treatment outcomes in 111 patients with septic diseases of the hand (septic arthritis, tendovaginitis, phlegmon). Surgical treatment was finished by primary sutures and drainage. They were divided into two groups. Bacteriophages were used in the main group. Other components of treatment were identical. «Pyobacteriophage complex¼ was injected into drainage tubes during dressings. Effectiveness of treatment was evaluated considering elimination of septic process, bacterial flora and healing of surgical wound. In addition, we performed in vitro testing of microorganisms for sensitivity to Pyobacteriophage. RESULTS: Staphylococcus and Streptococcus genus prevailed in hand infections. Pasteurella multocida has been isolated after pet bites. Growth of microflora in drainage tubes was observed in 16.7% and 21.1% of cases, respectively (p=0.02). Wound healing via primary intention was found in 83.6% and 71.4%, respectively (p=0.03). When testing in vitro, we observed lysis of cultures of Staphylococcus aureus in 83.6% of cases, Streptococcus pyogenes - in 33.3% of cases. CONCLUSION: «Pyobacteriophage complex¼ was effective for septic diseases of the hand. However, its partial lytic activity against the most common pathogens makes it advisable to combine phages and antibiotic therapy. Systematic renewal of phages is important due to widespread resistance of bacteria.

Ecological criteria for assessing the content of petroleum hydrocarbons in the main soils of coniferous-deciduous forests and forest steppe.

The effect of pollution of Albicluvisols/Retisols, Calcaric Leptosols, Luvic Phaeozems, Greyzamic Phaeozems and Folic Fluvisols with oil (Solovatovsky oil field, Perm region) added in amounts of 1, 2, 3 and 5 g oil/kg of soil on the organisms was studied in a model laboratory experiment. Oil addition showed phytotoxic effects on root length in Triticum aestivum L., Lepidium sativum L., Picea obovata Ledeb. and Pinus sylvestris L. in all soils. However, oil contamination of Calcaric Leptosols and Greyzamic Phaeozems led to growth stimulation in Picea obovata seedlings. A remarkable shift in the diversity and number of colony-forming units of heterotrophic and oil-oxidizing bacteria was detected in all soil types. The maximum decrease in biodiversity (45%) was noted for heterotrophic bacteria in Luvic Phaeozems. Aqueous extracts from all oil-contaminated soils had a toxic effect on Chlorella vulgaris Beijer, causing an increase in biomass by more than 30%, but did not show acute toxicity on Daphnia magna Straus. Oil addition in the range of 1-3 g oil/kg soil posed no environmental risk to human health. However, oil addition at 5 g oil/kg of soil led to an increase in the level of carcinogenic risk to children to the threshold values of acceptable risk and ranged from 0.95 × 10-4 for Greyzamic Phaeozems and Folic Fluvisols to 1.098 × 10-4 for Luvic Phaeozems. Our results suggest that the reaction of test organisms to oil pollution depends on the soil type, and their complex application makes it possible to identify the most sensitive factor and assess the dangerous level of pollution.

Nanobodies Are Potential Therapeutic Agents for the Ebola Virus Infection.

Ebola fever is an acute, highly contagious viral disease with a mortality rate that can reach 90%. There are currently no licensed therapeutic agents specific to Ebola in the world. Monoclonal antibodies (MAbs) with viral-neutralizing activity and high specificity to the Ebola virus glycoprotein (EBOV GP) are considered as highly effective potential antiviral drugs. Over the past decade, nanobodies (single-domain antibodies, non-canonical camelid antibodies) have found wide use in the diagnosis and treatment of various infectious and non-infectious diseases. In this study, a panel of nanobodies specifically binding to EBOV GP was obtained using recombinant human adenovirus 5, expressing GP (Ad5-GP) for alpaca (Vicugna pacos) immunization, for the first time. Based on specific activity assay results, affinity constants, and the virus-neutralizing activity against the recombinant vesicular stomatitis virus pseudotyped with EBOV GP (rVSV-GP), the most promising clone (aEv6) was selected. The aEv6 clone was then modified with the human IgG1 Fc fragment to improve its pharmacokinetic and immunologic properties. To assess the protective activity of the chimeric molecule aEv6-Fc, a lethal model of murine rVSV-GP infection was developed by using immunosuppression. The results obtained in lethal model mice have demonstrated the protective effect of aEv6-Fc. Thus, the nanobody and its modified derivative obtained in this study have shown potential protective value against Ebola virus.

[The pharmacoeconomic efficacy of lurasidone in the treatment of schizophrenia]. / Farmakoekonomicheskaya effektivnost' primeneniya preparata Lurazidon pri lechenii shizofrenii.

OBJECTIVE: To conduct a comprehensive pharmacoeconomic evaluation of lurasidone for the treatment of patients with schizophrenia under Russian healthcare system conditions and inclusion in EDL (Essential Drugs List) and Medication List for the Certain Categories of Citizens. MATERIAL AND METHODS: A retrospective study of lurasidone in the treatment of patients with schizophrenia was performed. Methods of pharmacoeconomic analysis were: cost analysis, budget impact analysis and cost-effectiveness analysis. RESULTS: Use of lurasidone for the treatment of patients with schizophrenia requires 50.04% less costs than the use of paliperidone and 46.69% less costs than the use of sertindole allowing to provide additional therapies to 100.1 and 87.6% of patients, respectively. The cost minimization analysis results are stable when prices fluctuate in the range of ±30%. Considering the current volume of antipsychotic drug supply, replacing 100% of paliperidone with lurasidone from the first year will reduce the cost of antipsychotics for patients who received paliperidone by 39.79 or by 360.81 million rubles over 3 years. Replacing 100% of sertindole with lurasidone from the first year will reduce the cost of antipsychotics for patients who received sertindole by 37.21 or 173.87 million rubles over 3 years. The results of the budget impact analysis are resistant to changes in prices for compared drugs in a wide range. CONCLUSION: Lurasidone is a more effective drug for treatment of schizophrenia from a pharmacoeconomic point of view in comparison with paliperidone and sertindole. With comparative efficacy with paliperidone and sertindole the use of lurasidone can significantly reduce the burden on budget of state programs of compensation for certain categories of citizens.

Carcinogenic and teratogenic status of human population and polychlorinated biphenyls contaminations of soils and biota (European pied flycatcher) in a Perm (Western Ural, Russia).

Polychlorinated biphenyls (PCBs) are included in the persistent organic pollutants designated by the Stockholm Convention and are hazardous compounds both for the environment and public health. The aim of this study was to determine the level of environmental pollution of PCBs in the Perm, in soil and biota (European pied flycatcher), and to analyze whether its presence was the cause of cancer and congenital malformations in the population. Soils in the study area had PCB concentrations of 101.87 µg/kg in the Industrialniy District and 27.81 µg/kg in the Leninskiy District of the Perm in 2005. The chemical composition of the individual PCBs and PCB groups were the same in the soils of both regions. The blood of nestlings of the European pied flycatcher raised in the Industrialniy District contained 9.61 ng PCB/ml, while those in the Leninskiy District had 5.64 ng PCB/ml in 2005. A linear correlation was established between the PCB contamination of soils and PCB contamination of pied flycatcher nestling's blood, and inverse linear correlation was established between the content of PCB in the blood of nestlings and the success of breeding of the pied flycatcher in Perm. An epidemiological analysis revealed a high incidence of cancer among the human population of the Industrialniy and Leninskiy districts (371.7 and 376.85 cases per 100,000 population, mean for the 2003-2018, respectively), which exceeded the figure for the whole of Perm (350.77 cases per 100,000 population, mean for the 2003-2018). The incidence rate of congenital malformations in Perm for the study period was 48.51 per 1000 human births. However, a decrease in the concentration of PCBs in soil and biota over a 15-year period (2005-2019) to the less than the detection limit did not lead to a decrease in the incidence of these diseases. Probably, PCB contamination was not the main cause of oncological diseases and congenital malformations in the population of the study area.

[Elimination of discoordination of the masticatory muscles work in patients with muscular-articular dysfunction of the temporomandibular joint by using «elastocorrector¼ appliance]. / Primenenie elastokorrektora dlya ustraneniya diskoordinatsii raboty zhevatel'nykh myshts u patsientov s disfunktsiei visochno-nizhnechelyustnogo sustava.

The aim of the study was to analyze the effectiveness of elastocorrective therapy in the complex treatment of patients with musculo-articular dysfunction of TMJ. Patients (n=237) underwent a comprehensive examination, including: 1) analysis of control and diagnostic plaster models of the jaws; 2) clinical examination; 3) X-ray examination: panoramic X-ray and TRG in lateral projection, computed tomography (CT) of the temporomandibular joint or X-ray zonography of the temporomandibular joint, and magnetic resonance tomography (MRI) of the temporomandibular joint, 4) electromyography of the chewing muscles, 5) T-Scan occlusion computer analysis; 6) stabilometry. After clinical, X-ray examination and data analysis of functional research methods, an algorithm for diagnostic and therapeutic measures was developed. All patients were prescribed the use of elastocorrector: at night during sleep and during the day for 1.5-2 hours. As a result of the use of elastocorrector in patients with muscle-articular dysfunction of the TMJ the position of the mandible was normalized and the spatial ratio of TMJ elements was optimized, the masticatory muscles function and bioelectric activity was normalized and the overload of muscles of mastication during functional movements is eliminated. «Elastocorrector¼ appliance can be used in patients with TMJ musculo-articular dysfunction both for orthodontic treatment (to eliminate chewing muscle hypertonus, compression of intra-articular structures, premature occlusive contacts) and to prepare for further treatment (orthopedic, therapeutic, etc.).

[X-ray assessment of the position of the lower jaw heads in different types of face]. / Rentgenologicheskaia otsenka polozheniia golovok nizhnei cheliusti pri razlichnykh tipakh litsa.

AIM: To increase the efficiency of diagnosis and orthodontic treatment of patients with temporomandibular joint dysfunction based on the study of the relationship between the features of the position of the heads of the lower jaw and various face types in patients aged 25 to 44 years according to cone beam computed tomography. We examined 100 people who were divided into two main groups (50 males, 50 females). To study the formed groups index method for diagnosing facial types according to Isar was used. The facial indicator was determined: the percentage of physiognomic height to the morphological width of the face. The structural features and relative positions of the TMJ bone structures in patients with TMJ dysfunction in various face types were studied in the Avantis 3D computer program. It was noted that the upper position of the lower jaw heads in women and men with a narrow type of face was observed in 50% and 40% of cases, respectively; the distal position of the lower jaw heads in women and men with a wide type of face was observed in 50% and 42.3% of cases, respectively. The results of the study indicate that there is a relationship between different types of face and the position of the lower jaw heads in the lower jaw, which should be considered in comprehensive treatment planning. The most at risk of temporal dysfunction is the TMJ of a woman with a narrow type of face, in which dominates the upper position of the lower jaw heads and a woman with a wide type of face with the distal position of the lower jaw heads. Before orthodontic treatment it is necessary to investigate the position of the TMJ heads with the help of CBCT and make a mandatory correction before the beginning of orthodontic treatment.

[Redox-dependent mechanisms of regulation of breast epithelial cell proliferation]. / Redoks-zavisimye mekhanizmy reguliatsii proliferatsii kletok épiteliia molochnoi zhelezy.

Activation of free radical oxidation in different cell types, including breast epithelial cells, may result in damage to macromolecules, in particular, proteins taking part in regulation of cell proliferation and apoptosis. The glutathione, glutaredoxin and thioredoxin systems play an essential role in maintaining intracellular redox homeostasis. Due to this fact, modulation of cellular redox status under the effect of an SH group inhibitor and an SH group protector may be used as a model for studying the role of redox proteins and glutathione in regulating cell proliferation in different pathological processes. In this study we have evaluated the state of the thioredoxin, glutaredoxin and glutathione systems as well as their role in regulating proliferation of HBL-100 breast epithelial cells under redox status modulation with N-ethylmaleimide (NEM) and 1,4-dithioerythriol (DTE). Modulating the redox status of breast epithelial cells under the effect of NEM and DTE influences the functional activity of glutathione-dependent enzymes, glutaredoxin, thioredoxin, and thioredoxin reductase through changes in the GSH and GSSG concentrations. In HBL-100 cells under redox-status modulation, we have found an increase in the number of cells in the S-phase of the cell cycle and a decrease in the number of cells in the G0/G1 and G2/М phases, as opposed to the values in the intact culture. The proposed model of proliferative activity of cells under redox status modulation may be used for development of new therapeutic approaches for treatment of diseases accompanied by oxidative stress generation.

Safety and immunogenicity of GamEvac-Combi, a heterologous VSV- and Ad5-vectored Ebola vaccine: An open phase I/II trial in healthy adults in Russia.

Ebola hemorrhagic fever, also known as Ebola virus disease or EVD, is one of the most dangerous viral diseases in humans and animals. In this open-label, dose-escalation clinical trial, we assessed the safety, side effects, and immunogenicity of a novel, heterologous prime-boost vaccine against Ebola, which was administered in 2 doses to 84 healthy adults of both sexes between 18 and 55 years. The vaccine consists of live-attenuated recombinant vesicular stomatitis virus (VSV) and adenovirus serotype-5 (Ad5) expressing Ebola envelope glycoprotein. The most common adverse event was pain at the injection site, although no serious adverse events were reported. The vaccine did not significantly impact blood, urine, and immune indices. Seroconversion rate was 100 %. Antigen-specific IgG geometric mean titer at day 42 was 3,277 (95 % confidence interval 2,401-4,473) in volunteers immunized at full dose. Neutralizing antibodies were detected in 93.1 % of volunteers immunized at full dose, with geometric mean titer 20. Antigen-specific response in peripheral blood mononuclear cells was also detected in 100 % of participants, as well as in CD4+ and CD8+ T cells in 82.8 % and 58.6 % of participants vaccinated at full dose, respectively. The data indicate that the vaccine is safe and induces strong humoral and cellular immune response in up to 100 % of healthy adult volunteers, and provide a rationale for testing efficacy in Phase III trials. Indeed, the strong immune response to the vaccine may elicit long-term protection. This trial was registered with grls.rosminzdrav.ru (No. 495*), and with zakupki.gov.ru (No. 0373100043215000055).

Long-lived coherence in pentafluorobenzene as a probe of ππ(*) - πσ(*) vibronic coupling.

The dynamics of pentafluorobenzene after femtosecond laser excitation to the optically bright ππ(*) first excited electronic state have been investigated by femtosecond time-resolved time-of-flight mass spectrometry and femtosecond time-resolved photoelectron imaging spectroscopy. The observed temporal profiles exhibit a bi-exponential decay behavior with a superimposed, long-lived, large-amplitude oscillation with a frequency of νosc = 78-74 cm(-1) and a damping time of τD = 5-2 ps. On the basis of electronic structure and quantum dynamics calculations, the oscillations have been shown to arise due to vibronic coupling between the optically bright ππ(*) state and the energetically close-lying optically dark πσ(*) state. The coupling leads to a pronounced double-well character of the lowest excited adiabatic potential energy surface along several out-of-plane modes of b1 symmetry. The optical electronic excitation initiates periodic wavepacket motion along these modes. In the out-of-plane distorted molecular configuration, the excited state acquires substantial πσ(*) character, thus modulating the ionization probability. The photoelectron spectra and the anisotropy of their angular distribution confirm the periodically changing electronic character. The ionizing probe laser pulse directly maps the coupled electron-nuclear motion into the observed signal oscillations.

[Polymorphism of the bphA genes in bacteria destructing biphenyl/chlorinated biphenils].

Polychlorinated biphenyls (PCBs) are persistent organic pollutants. Biphenyl 2,3-dioxygenase (BDO) is a key enzyme that determines the range of PCBs oxidized by a bacterial strain. BDO subunit α (BphA1) plays an essential role in substrate recognition and binding. The genes for dioxygenases that hydroxylate aromatic rings were screened and analyzed phylogenetically. Genes found in biphenyl-oxidizing Rhodococcus erythropolis strains G12a, B7b, and B106a proved to be similar to the published nucleotide sequences of the Rhodococcus sp. HA99 and R04 and Novosphingobium aromaticivorans F199 bphA1 genes, which code for the α-subunits that do not belong to the biphenyl/toluene dioxygenase (B/TDO) family. PCB-destructing R. ruber P25 was found to possess a unique bphA1 gene, which clusters together with the phenylpropionate dioxygenase (PPDO) α-subunits of Mycobacterium vanbaalenii PYR-1 and Frankia sp. EuI1c. The deduced amino acid sequences of the genes were analyzed. The amino acids of the BDO active site in R. wratislaviensis P1, P12, P13, and P20 (bphA1 genes of the B/TDO family) were identical to those of the active PCB degrader R. jostii RHA1. The Rhodococcus strains in question were shown to be active toward both orthoand parachlorinated ring of 2,4'-dichlorobiphenyl. The α-subunit amino acids responsible for the substrate specificity of the enzyme in Pseudomonas sp. S9, S13, S210, S211, and S212 (B/TDO family) were the same as in P. pseudoalcaligenes KF707. The Pseudomonas strains were active toward the para-chlorinated ring of 2,4'-dichlorobiphenyl. The results of screening bacterial strains for bphA1 can be used to identify the biotechnologically promising PCB destructors.

[Molecular biological characterization of biphenyl-degrading bacteria and identification of the biphenyl 2,3-dioxygenase α-subunit genes].

Bacterial isolates from soils contaminated with (chlorinated) aromatic compounds, which degraded biphenyl/chlorinated biphenyls (CB) and belonged to the genera Rhodococcus and Pseudomonas were studied. Analysis of the 16S rRNA gene sequences was used to determine the phylogenetic position of the isolates. The Rhodococcus cells were found to contain plasmids of high molecular mass (220-680 kbp). PCR screening for the presence of the bphA1 gene, a marker indicating the possibility for induction of 2,3-dioxygenase (biphenyl/toluene dioxygenase subfamily) revealed the presence of the bphAl genes with 99-100% similarity to the homologous genes of bacteria of the relevant species in all pseudomonad and most Rhodococcus isolates. A unique bphA1 gene, which had not been previously reported for the genus, was identified in Rhodococcus sp. G10. The absence of specific amplification of the bphA1 genes in some biphenyl-degrading bacteria (Rhodococcus sp. B7b, B106a, G12a, P2kr, P2(51), and P2m), as well as in an active biphenyl degrader Rhodococcus ruber P25 indicated the absence of the genes encoding the proteins of the biphenyl/toluene dioxygenase subfamily and participation of the enzymes other than this protein family in biphenyl/CB degradation.

Bioaugmentation of a polychlorobiphenyl contaminated soil with two aerobic bacterial strains.

The consortium of aerobic bacterial strains Rhodococcus ruber P25 and Microbacterium sp. B51 was bioaugmented in natural and industrial soils, contaminated by commercial mixture of polychlorinated biphenyls (PCBs) Sovol. The results showed that the bioaugmentation of bacterial strains led to PCBs degradation in soil. Sovol at the initial concentration of about 100 mg kg(-1) was removed by 72.2% in the bioaugmented system with natural soil within 90 days, while the system with industrial soil removed 96.4% of this compound within the same period. The biodegradation kinetics of PCBs in the bioaugmented soil systems was not dependent on the presence of indigenous microflora. It was found that the growth dynamics of the strains R. ruber P25 and Microbacterium sp. B51 correlated with the specific degradation of Sovol. The strains R. ruber P25 and Microbacterium sp. B51 displayed high degradative activity to all congeners (ortho-, meta- and para-substituent) contained in Sovol. Removal percentage for each congeners amounted to 59-100% in the bioaugmented systems. This study suggests that augmentation of PCB-contaminated soils with strain R. ruber P25 and Microbacterium sp. B51 is promising in PCB bioremediation.

[Destruction of aromatic hydrocarbons by the Rhodococcus wratislaviensis KT112-7 strain isolated from waste products of a salt-mining factory].

The destruction of aromatic hydrocarbons by the Rhodococcus wratislaviensis KT112-7 strain isolated from technogenic mineral waste products of the BKRU1 Uralkalii factory has been investigated (city of Berezniki, Perm krai). The R. wratislaviensis KT112-7 was shown to utilize increased concentrations of ophthalic (o-PA) (8 g/L) and benzoic (BA) (3.4 g/L) acids. The strain grows with o-FA, BA, and biphenyl at a NaCl content of up to 50, 90, and 75 g/L in the culture medium, respectively. Based on an analysis of the metabolic profile and nucleotide sequences of the bphA1, benA, and phtB genes, the KT112-7 strain was established to decompose o-PA via the formation of 3,4-dihydroxyphthalic and 3,4-dihydroxybenzoic acids. The decomposition of biphenyl is carried out via the formation of BA and then at low concentrations of NaCl (up to 50 g/L) via the formation of 4-hydroxybenzoic acid followed by its oxidation; at high concentrations of NaCl (over 60 g/L), via the direct oxidation of benzoic acid with the production of catechol. These data indicate that the Rhodococcus wratislaviensis KT112-7 destructor strain is a promising strain for the development of new biotechnologies directed at the utilization (transformation) of aromatic compounds, including under the conditions of increased mineralization.

[Destruction of mixture of tri-hexa-chlorinated biphenyls by Rhodococcus genus strains].

Destruction of polychlorinated biphenyls (PCBs) by strain-destructors Rhodococcus sp. B7a and Rhodococcus sp. G12a has been studied. It was shown that these strains destruct 78-95% of PCB mixture containing tri-hexa-chlorinated biphenyls. Rhodococcus destruct all components of the mixture of tri-, tetra-, penta-, and hexa-chlorinated biphenyls without accumulation of toxic chlorinated metabolites. The studied bacteria destruct PCB that are the most stable for oxidation, such as 2,5,2',5'-CB; 3,4,3',4'-CB; and 2,4,5,2',4',5'-CB. The most perspective strains are R. rubber P25, Rhodococcus sp. B7a and Rhodococcus sp. G12a whose metabolic potential can be used for biotechnological refinement of the environment from highly toxic pollutants.

[Degradation of chlorinated biphenyls and products of their bioconversion by Rhodococcus sp. B7a strain].

Strain Rhodococcus sp. B7a isolated from artificially polluted soil destructs mono- and di-substituted ortho- and/or para-chlorinated biphenyls with utilization of chlorinated benzoic acids and shows high degradation activity as regards trichlorinated biphenyls. It is shown that p-hydroxybenzoic and protocatehoic acids are the products of p-chlorobenzoic acid catabolism.