[Identification of Melittin-Like Proteins with a Molecular Weight of 67 κDa that Interact with Na

Melittin, a peptide from bee venom, was found to be able to interact with many proteins, including calmodulin target proteins and ion-transporting P-type ATPases. It is assumed that melittin mimics a protein module involved in protein-protein interactions within cells. Previously, a Na^(+)/K^(+)-ATPase containing the α1 isoform of the catalytic subunit was found to co-precipitate with a protein with a molecular weight of about 70 κDa that interacts with antibodies against melittin by cross immunoprecipitation. In the presence of a specific Na^(+)/K^(+)-ATPase inhibitor (ouabain), the amount of protein with a molecular weight of 70 κDa interacting with Na^(+)/K^(+)-ATPase increases. In order to identify melittin-like protein from murine kidney homogenate, a fraction of melittin-like proteins with a molecular weight of approximately 70 κDa was obtained using affinity chromatography with immobilized antibodies specific to melittin. By mass spectrometry analysis, the obtained protein fraction was found to contain three molecular chaperones of Hsp70 superfamily: mitochondrial mtHsp70 (mortalin), Hsp73, Grp78 (BiP) of endoplasmic reticulum. These data suggest that chaperones from the HSP-70 superfamily contain a melittin-like module.

[Metabolic Stress of Red Blood Cells Induces Hemoglobin Glutathionylation].

Metabolic stress caused by a lack of glucose significantly affects the state of red blood cells, where glycolysis is the main pathway for the production of ATP. Hypoglycemia can be both physiological (occurring during fasting and heavy physical exertion) and pathological (accompanying a number of diseases, such as diabetes mellitus). In this study, we have characterized the state of isolated erythrocytes under metabolic stress caused by the absence of glucose. It was established that 24 h of incubation of the erythrocytes in a glucose-free medium to simulate blood plasma led to a two-fold decrease in the ATP level into them. The cell size, as well as intracellular sodium concentration increased. These findings could be the result of a disruption in ion transporter functioning because of a decrease in the ATP level. The calcium level remained unchanged. With a lack of glucose in the medium of isolated erythrocytes, there was no increase in ROS and a significant change in the level of nitric oxide, while the level of the main low-molecular weight thiol of cells, glutathione (GSH) decreased by almost 2 times. It was found that the metabolic stress of isolated red blood cells induced hemoglobin glutathionylation despite the absence of ROS growth. The cause was the lack of ATP, which led to a decrease in the level of GSH because of the inhibition of its synthesis and, probably, due to a decrease in the NADPH level required for glutathione (GSSG) reduction and protein deglutathionylation. Thus, erythrocyte metabolic stress induced hemoglobin glutathionylation, which is not associated with an increase in ROS. This may have an important physiological significance, since glutathionylation of hemoglobin changes its affinity for oxygen.

[Tumnin mineral spring. History of development]. / Tumninskii mineral'nyi istochnik. Istoriya osvoeniya.

The article dwells upon the history of the discovery of the Tumnin mineral spring, the establishment and development of the Far Eastern health resort «Goryachy Klyuch,¼ located in the basin of Chope creek, a tributary of the largest river in the eastern macroslope of Sikhote Alin, Tumnin river, located 25 km from the Strait of Tartary. A historical sketch since the first mentioning of the Tumnin mineral spring from 1903 to the present day, as well as the results of hydrogeological expeditions to determine the chemical composition and α-activity of Tumnin mineral water at different periods, are presented. A contribution of a geological expedition that established a large deep-lying tectonic structure permeable to upwelling thermal water flows is described. The role of the staff of the physiotherapy and balneology department of the Khabarovsk Medical Institute in the study of the mechanism of action and clinical effectiveness of the Tumnin mineral water is addressed. A balneological characteristic of nitric and siliceous thermal water, the basic therapeutic factor of «Goryachy Klyuch¼ health resort, which has always been popular among the Far East residents, but gained special importance and appreciation of patients during the pandemic of new coronavirus infection, is given. Currently, in the health resort «Goryachy Klyuch¼, patients with skin diseases, musculoskeletal, gynecologic, neurologic diseases, digestive tract disorders, metabolic conditions, upper airways, cardiovascular disorders, occupational diseases are treated using balneotherapy and other methods of non-drug therapy. At present, the health resort «Goryachy Klyuch¼ is going through a difficult but interesting period of improvement of recreation opportunities for the Far East residents.

Na+ i/K+ i imbalance contributes to gene expression in endothelial cells exposed to elevated NaCl.

High-salt consumption contributes to the development of hypertension and is considered an independent risk factor for vascular remodelling, cardiac hypertrophy and stroke incidence. Alterations in NO production, inflammation and endothelial cell stiffening are considered now as plausible mediators of cardiovascular dysfunction. We studied early responses of endothelial cells (HUVEC) caused by a moderate increase in extracellular sodium concentration. Exposure of HUVEC to elevated sodium within the physiological range up to 24 h is accompanied by changes in monovalent cations fluxes and Na,K-ATPase activation, and, in turn, results in a significant decrease in the content of PTGS2, IL6 and IL1LR1 mRNAs. The expression of NOS3 and FOS genes, as well as the abundance of cytosolic and nuclear NFAT5 protein, remained unchanged. We assessed the mechanical properties of endothelial cells by estimating Young's modulus and equivalent elastic constant using atomic force and interference microscopy, respectively. These parameters were unaffected by elevated-salt exposure for 24 h. The data obtained suggest that even small and short-term elevations of extracellular sodium concentration affect the expression of genes involved in the control of endothelial function through the Na+ i/K+ i-dependent mechanism(s).

[Comparative efficacy of various treatment regimens for children 2-5 years old with symptoms of acute viral rhinosinusitis]. / Sravnitel'naya effektivnost' razlichnykh skhem lecheniya detei 2­5 let s simptomatikoi ostrogo virusnogo rinosinusita.

In an open, randomized prospective comparative clinical study in parallel groups of patients with a diagnosis of acute respiratory infection, acute rhinosinusitis, 107 children aged 4-5 years took part. The purpose of this study was to compare the effectiveness of various treatment regimens for acute viral rhinosinusitis in children. We compared the treatment effectiveness of acute viral rhinosinusitis using the herbal medicine Sinupret in the form of oral drops, standard treatment and standard treatment supplemented with topical antibacterial drugs. The objectivity of treatment effectiveness evaluation was based on the complaints' dynamics, the changes' presence in the mucous membrane of the nasal cavity within 10 days: at the initial visit, after 3 days, on the 7th and 10th days after the initial examination. A follow-up visit was also scheduled for the 14th day. The analysis of the obtained data revealed that the Sinupret usage in the complex treatment of children with viral etiology acute rhinosinusitis contributes to a more pronounced positive symptoms' dynamics compared to children who did not receive Sinupret. Sinupret eliminates mucostasis, which contributes to a faster restoration of the drainage and ventilation function of the auditory tube, restores mucociliary clearance, improves the condition and well-being of children, improves the effectiveness of therapy and shortens the treatment time.

The use of 4-Hexylresorcinol as antibiotic adjuvant.

Ever decreasing efficiency of antibiotic treatment due to growing antibiotic resistance of pathogenic bacteria is a critical issue in clinical practice. The two generally accepted major approaches to this problem are the search for new antibiotics and the development of antibiotic adjuvants to enhance the antimicrobial activity of known compounds. It was therefore the aim of the present study to test whether alkylresorcinols, a class of phenolic lipids, can be used as adjuvants to potentiate the effect of various classes of antibiotics. Alkylresorcinols were combined with 12 clinically used antibiotics. Growth-inhibiting activity against a broad range of pro- and eukaryotic microorganisms was determined. Test organisms did comprise 10 bacterial and 2 fungal collection strains, including E. coli and S. aureus, and clinical isolates of K. pneumoniae. The highest adjuvant activity was observed in the case of 4-hexylresorcinol (4-HR), a natural compound found in plants with antimicrobial activity. 50% of the minimal inhibitory concentration (MIC) of 4-HR caused an up to 50-fold decrease in the MIC of antibiotics of various classes. Application of 4-HR as an adjuvant revealed its efficiency against germination of bacterial dormant forms (spores) and prevented formation of antibiotic-tolerant persister cells. Using an in vivo mouse model of K. pneumoniae-induced sepsis, we could demonstrate that the combination of 4-HR and polymyxin was highly effective. 75% of animals were free of infection after treatment as compared to none of the animals receiving the antibiotic alone. We conclude that alkylresorcinols such as 4-HR can be used as an adjuvant to increase the efficiency of several known antibiotics. We suggest that by this approach the risk for development of genetically determined antibiotic resistance can be minimized due to the multimodal mode of action of 4-HR.

[Kolyma health resort «Talaya¼ (to its 80th birthday)]. / Kolymskaya zdravnitsa «Talaya¼. K 80-letiyu.

The article is devoted to the history of the creation and establishment of the Far Eastern resort «Talaya¼, located in the Far North of Russia, in the Magadan Region among the hills of the Kolyma Range, in the valley of the Talaya River. A historical essay is presented from 1868 to the present days, including periods of the resort's development at various stages of Russia's formation (pre-revolutionary, post-revolutionary periods, the era of developed socialism, present times). The contribution of resort doctors, geologists, employees of research institutes to the study of the mechanism of action and clinical effectiveness of the use of Tal mineral water is described. The balneological characteristic of nitrogen-siliceous thermal water, the main therapeutic factor of the Talaya resort, is given. Currently, the Talaya sanatorium is providing the treatment of patients with diseases of the skin, musculoskeletal system, gynecological, neurological diseases, diseases of the digestive system, metabolism, upper respiratory tract, cardiovascular system, occupational diseases using balneotherapy, mud therapy and others methods of non-drug therapy. Hundreds of thousands of northerners were healed by the Kolyma health resort, which gained fame as the «Northern Pearl¼. Today, during the reorganization of health care, the Talaya sanatorium is going through a difficult but interesting period in improving the possibilities of healing the inhabitants of the northern regions of Russia.

Effects of Dynamic and Static Loads on the Concentration of Sodium and Potassium in Murine Skeletal Muscles.

The effects of dynamic and static load on the intracellular concentrations of sodium [Formula: see text] and potassium (K+i) in m. soleus and m. biceps, respectively, were studied in mice. Both dynamic (swimming for 60 min) and static (hanging on the grid for 40 min) load led to a 2-fold increase in [Formula: see text] level, a decrease in K+i concentration by 25-35%, and 3-4-fold increase in the [Formula: see text] ratio. These effects of dynamic and static loads on the studied parameters remained unchanged in mice subjected to regular physical exercise (swimming or hanging on the grid for 1 h a day over 4 weeks). Our results suggest that dissipation of sodium and potassium transmembrane gradients during physical exercise can be considered as a factor of regulation of functional activity of skeletal muscles, which includes changes in transcription and translation of myokines observed previously.

Ouabain and Marinobufagenin: Physiological Effects on Human Epithelial and Endothelial Cells.

Long-term study on the identification of Na,K-ATPase endogenous inhibitors in mammalian tissues has resulted in the discovery of ouabain, marinobufagenin (MBG), and other cardiotonic steroids (CTS) in the blood plasma. Production of ouabain and MBG is increased in essential hypertension and other diseases associated with hypervolemia. Here, we compared the effects of ouabain and MBG on the Na,K-ATPase activity (measured as the transport of Na+, K+, and Rb+ ions) and proliferation and death of human renal epithelial cells (HRECs) and human umbilical vein endothelial cells (HUVEC) expressing α1-Na,K-ATPase. Ouabain concentration that provided the half-maximal inhibition of the Rb+ influx (IC50) into HRECs and HUVECs was 0.07 µM. In both types of cells, the IC50 values for MBG were 10 times higher than for ouabain. Incubation of HREC and HUVEC with 0.001-0.01 µM ouabain for 30 h resulted in 40% increase in the [3H]thymidine incorporation into DNA; further elevation of ouabain concentration to 0.1 µM completely suppressed DNA synthesis. MBG at the concentration of 0.1 µM activated DNA synthesis by 25% in HRECs, but not in HUVECs; 1 µM MBG completely inhibited DNA synthesis in HRECs and by 50% in HUVECs. In contrast to HRECs, incubation of HUVECs in the serum-free medium induced apoptosis, which was almost completely suppressed by ouabain and MBG at the concentrations of 0.1 and 3 µM, respectively. Based on these data, we can conclude that (i) the effect of MBG at the concentrations detected in the blood plasma (<0.01 µM) on HRECs and HUVECs was not due to the changes in the [Na+]i/[K+]i ratio; (ii) the effect of physiological concentrations of ouabain on these cells might be mediated by the activation of Na,K-ATPase, leading to cell proliferation.

[Balneological resource of the 'Zharkie Vody' (Hot Waters) spring (Iturup Island)]. / Bal'neologicheskii resurs istochnika 'Zharkie vody' (o. Iturup).

BACKGROUND: It is necessary to develop a balneotherapeutic and tourist-recreational complex on Iturup Island (hereinafter referred to as Iturup Island). This investigation was conducted in the framework of the federal target program 'Socioeconomic development of the Kuril Islands, Sakhalin Region, for 2007-2015' under No. 478 dated 08.09.06. OBJECTIVE: To characterize the balneological resource of the 'Zharkie Vody' (Hot Water) spring on Iturup Island. MATERIAL AND METHODS: Expeditionary studies were conducted at the 'Zharkie Vody' spring located near the village of Raidovo (Iturup Island): its water samples were taken and chemically analyzed in a certified laboratory in Khabarovsk; the biological activity of the mineral water was studied using the Cyto-Expert portable non-invasive diagnostic equipment adapted to the expeditionary conditions. RESULTS: The mineral water from the 'Zharkie Vody' deposit (Iturup Island) is weakly mineralized siliceous sulfate-bicarbonate-chloride calcium-sodium thermal and has an activation function on living cell structures. CONCLUSION: The data obtained from the studies created the prerequisites for setting up a balneotherapy complex near the 'Zharkie Vody' on Iturup Island and provide optimistic forecasts for the further study of this deposit.

Search for Intracellular Sensors Involved in the Functioning of Monovalent Cations as Secondary Messengers.

Maintenance of non-equilibrium Na+ and K+ distribution between cytoplasm and extracellular medium suggests existence of sensors responding with conformational transitions to the changes of these monovalent cations' intracellular concentration. Molecular nature of monovalent cation sensors has been established in Na,K-ATPase, G-protein-coupled receptors, and heat shock proteins structural studies. Recently, it was found that changes in Na+ and K+ intracellular concentration are the key factors in the transcription and translation control, respectively. In this review, we summarize results of these studies and discuss physiological and pathophysiological significance of Na+i,K+i-dependent gene expression regulation mechanism.

A mathematical model for predicting the development of bacterial resistance based on the relationship between the level of antimicrobial resistance and the volume of antibiotic consumption.

OBJECTIVES: Infections that are inadequately treated owing to acquired bacterial resistance are a leading cause of mortality. Rates of multidrug-resistant bacteria are rising, resulting in increased antibiotic failures and worsening patient outcomes. Mathematical modelling makes it possible to predict the future spread of bacterial antimicrobial resistance. The aim of this study was to construct a mathematical model that can describe the dependency between the level of antimicrobial resistance and the amount of antibiotic usage. METHODS: After reviewing existing mathematical models, a cross-sectional, retrospective study was carried out to collect clinical and microbiological data across 3000 patients for the construction of the mathematical model. Based on these data, a model was developed and tested to determine the dependency between antibiotic usage and resistance. RESULTS: Consumption of inhibitor/cephalosporins and fluoroquinolones increases inhibitor/penicillin resistance. Consumption of inhibitor/penicillins increases cephalosporin resistance. Consumption of inhibitor/penicillins increases inhibitor/cephalosporin resistance. CONCLUSIONS: It was demonstrated that in some antibiotic-micro-organism pairs, the level of antibiotic usage significantly influences the level of resistance. The model makes it possible to predict the change in resistance and also shows the quantitative effect of antibiotic consumption on the level of bacterial resistance.

Effects of Ouabain on Proliferation of Human Endothelial Cells Correlate with Na+,K+-ATPase Activity and Intracellular Ratio of Na+ and K.

Side-by-side with inhibition of the Na+,K+-ATPase ouabain and other cardiotonic steroids (CTS) can affect cell functions by mechanisms other than regulation of the intracellular Na+ and K+ ratio ([Na+]i/[K+]i). Thus, we compared the dose- and time-dependences of the effect of ouabain on intracellular [Na+]i/[K+]i ratio, Na+,K+-ATPase activity, and proliferation of human umbilical vein endothelial cells (HUVEC). Treatment of the cells with 1-3 nM ouabain for 24-72 h decreased the [Na+]i/[K+]i ratio and increased cell proliferation by 20-50%. We discovered that the same ouabain concentrations increased Na+,K+-ATPase activity by 25-30%, as measured by the rate of (86)Rb(+) influx. Higher ouabain concentrations inhibited Na+,K+-ATPase, increased [Na+]i/[K+]i ratio, suppressed cell growth, and caused cell death. When cells were treated with low ouabain concentrations for 48 or 72 h, a negative correlation between [Na+]i/[K+]i ratio and cell growth activation was observed. In cells treated with high ouabain concentrations for 24 h, the [Na+]i/[K+]i ratio correlated positively with proliferation inhibition. These data demonstrate that inhibition of HUVEC proliferation at high CTS concentrations correlates with dissipation of the Na+ and K+ concentration gradients, whereas cell growth stimulation by low CTS doses results from activation of Na+,K+-ATPase and decrease in the [Na+]i/[K+]i ratio.

Molecular Mechanisms of Ceftaroline Susceptibility Reduction in Methicillin-Resistant Staphylococcus aureus.

Ceftaroline is a unique cephalosporin with activity against methicillin resistant Staphylococcus aureus (MRSA). It was approved for clinical use in the USA, Europe and Russian Federation since 2010 for the treatment of the skin and soft tissue infection and community-acquired pneumoniae. In the present study there was used molecular typing of 24 isolates of MRSA with reduced susceptibility to ceftaroline. For 8 isolates belonging to different genetic lines (ST8, ST239 and ST228) and requiring MICs there were determined antibiotic concentrations preventing formation of resistant mutants (mutant prevention concentration) and the ranges of the mutant selection window (MSW). The last majority of the isolates with reduced susceptibility to ceftaroline (MIC of 2 mcg/ml) belonged to the clonal line ST228. The whole genome sequencing of two isolates of ST228 showed that they belonged to the epidemic South Germany genetic line and were characterized by the presence of mutations in PBP2a (N146K) and PBP2 (C197Y) responsible for reduced susceptibility. The highest rates of MPC (32 mcg/ml) and MSW (2-16 mcg/ml) were observed in the clinical isolates belonging to the genetic line ST8. The isolates of ST239 and ST228 had the selection window within 2-4 mcg/ml. No dependence of the MIC and MPC/MSW levels was detected.

SNP Polymorphism in Genomes of CC320 Isolates of Streptococcuspneumoniae Resistant to Beta-Lactams.

Bioinformatic analysis of the data on the genome sequencing of the isolates of the Streptococcuspneumoniae clonal complex SS320 from the Russian Federation, as well as the data on SS320 isolates from public sources in the penicillin resistant isolates resulted in detection of 139 missense mutations in 45 genes. In addition to the mutations in the genes of the main penicillin-binding proteins (PSB - PBP1A, PBP2B and PBP2X) there was detected high frequency of mutations in the genes of the (division and cell wall) dcw-cluster, as well as in RegR protein belonging to the transcription regulators of the LacI/GaIR family. Development of resistance to beta-lactams in S.pneumoniae is defined not only by modification of the PSB, but also by adaptive changes in the metabolic pathways involved in the bacterial cell growth and division.

Prevalence and Antibiotic Resistance of Carbapenemase-Producing Gram-Negative Bacteria in Saint Petersburg and Some Other Regions of the Russian Federation.

Carbapenemase-producing gramnegative bacteria, which hydrolyze most offi-lactams, including carbapenems, is of global health care system threat. The number of the known carbapenemases is constantly increasing, however only four types are widely distributed: NDM-type, KPC-type, OXA-48-type and VIM-type. The frequency of carbapenemase-producing Klebsiellapneumoniae in hospitals of Saint Petersburg reached 9.2% (5.9% for NDM-type, 1.4% for OXA-48-type, 1.9% for NDM-type + OXA-48-type). Carbapenemase producers were also detected in hospitals of Moscow, Yekaterinburg, Vologda, Murmansk, Kurgan, Krasnoyarsk, Izhevsk, Krasnodar and Perm. In total 281 carbapenemase producers were recorded within 2011-2016, which were isolated from infected or colonized patients (K.pneumoniae - 247 isolates, Acinetobacter spp - 29 isolates, Enterobacter cloacae - 2 isolates, Serratia marcescens - 1 isolate, Escherichia coli - 1 isolate and Proteus mirabifis - 1 isolate). The carbapenemase-producing K.pneumoniae isolates were distinguished by considerable genetic diversity, the NDM-type carbapenemase-producers belonged to eight, KPC-type - to three and OXA-48-type - to four different sequence-types (STs) respectively. The representatives of the globally dominant genetic line, Clonal Group 258 (CG258), and also a number of the less common lines (ST147, ST273, ST307 and ST377) were detected. The K.pneumoniae strains were distinguished by a high frequency of cross-resistance and the associated resistance to antibiotics of different groups. The frequency of resistance to cephalosporins and fluoroquinolones reached 100%. Among the NDM-type carbapenemase producers the frequency of resistance to aminoglycosides exceeded 90%, among the KPC-type carbapenemase producers the frequency of resistance corresponded to 66% for amikacin and 93% for gentamicin, among the OXA-48 type carbapenemase producers the frequency of resistance was even lower (50% and 73% respectively). Approximately 80% of the NDM-type, 90% of the KPC-type and only 60% of the OXA-48-type carbapenemase producers showed a high level of resistance to imipenem and meropenem. The frequency of resistance to tigecycline varied within 6.7% to 14.8% and the frequency of resistance to polymyxin was within 4.2% to 20%. The OXA-40- and OXA-23-types carbapenemase-producing Acinetobacter spp. remained susceptible only to polymyxin. It is obvious that the possibility of antibacterial therapy of infections caused by carbapenemases producers is limited.

[Annenskie mineral waters: the history of discovery (the 150th anniversary of the balneotherapeutic centre spa "Annenskie waters")].

The present article is an attempt to create a complete picture of the history of foundation of the first Far East balneotherapeutic spa centre known as "Annenskie waters". This article brings together the legends and historical information concerning this locality dated since 1858 up to the present time. The main stages of the development of the medical centre are reviewed starting from the period of unorganized chaotic attendance to the creation of the modern comfortable spa facility, building new bathrooms, halls, dormitories, dining and auxiliary premises. The process of developing and extending indications for the medical application of Annenskie mineral waters is followed backward in the past. The first report on the clinical effectiveness of Annenskie waters is presented. The contribution made by several historical figures to the development of the balneotherapeutic centre is described.

[Antibiotic Resistance of MRSA in the Russian Federation].

The results of the multicentre trial on estimation of MRSA antibiotic susceptibility to 17 antibiotics are presented. 474 nonrepeting isolates of MRSA (mecA+), collected in 2011-2014 in 10 cities of the Russian Federation were used in the trial. The antibiotic susceptibility was determined by the method of serial microdilutions in broth with estimation of the MICs in accordance with the international standards CLSI 2014 and EUCAST 2014. The highest levels of the MRSA resistance were stated against ciprofloxacin--92%(MIC50 32 mcg/ml), gentamicin--85% (MIC50 128 mcg/ml), erythromycin--54% (MIC50 32-mcg/ml) and clindainycin - 45% (MIC50 0.03 mcg/ml), as well as against rifampicin--38% (MIC50 0.06 mcg/ml). The frequency of MRSA isolated at the vancomycin dose of 2 mcg/ml equaled 26%. No correlation of the decrease in susceptibility to vancomycin and rifampicin was observed. In 5% of MRSA isolated from infected surgical wounds in patients with bone infection or sepsis, there was observed a decrease in the susceptibility to ceftarolin (MIC 2-4 mcg/ml). Co-trimoxasole, fusidic acid (MIC50 0.06 mcg/ml) and mupirocin (MIC50 0.5 mcg/ml) showed high antibacterial activity, 93-98% of the isolates being susceptible to the drugs. No resistance to linezolid and tigecycline was detected. By the associate resistance spectrum, most of the MRSA isolates were characterized by resistance to drugs of 3-7 groups (56%). The phenotypes with simultaneous resistance to drugs of 8-10 groups amounted to 6%. As a whole, 70 variants of associate resistance combinations were detected.

[Antibiotic Resistance and Serotype Pattern of Streptococcus pneumoniae Isolated from Children in St. Petersburg in 2010-2013].

The surveillance of the serotype pattern and antibiotic resistance of S. pneumoniae in various geographical regions is required for the validity of rational etiotrophic therapy of pneumococcal infections and the choice of the optimal vaccines for their prophylaxis. 250 S. pneumoniae isolates from children with acute otitis or pneumonia and healthy carriers in St. Petersburg in 2010-2013 were investigated. The analysis of the serotype pattern of the pneumococci showed that 13-valent conjugate vaccine was the most active (86.1% of pneumococci causing pneumonia and 86.4% of pneumococci causing acute otitis). The isolates were higly resistant to beta-lactams and macrolides. By the EUCAST criteria, the decrease in the susceptibility to penicillin, cefotaxime, erythromycin and ceftarolin was observed in 32.4%, 14%, 33.2 and 6% of the isolates respectively. 22.4% of the isolates showed associate resistance to penicillin and erythromycin.. No resistance to moxifloxacin was detected. The frequency of resistance to tetracycline, co-trimoxasole and chloramphenicol in various patients ranged within 30-50%. The prevalence of the antibiotic resistance was mainly characteristic of the isolates serotypes 19A, 19F, 14 and serogroup 6.

[Discovery of Genetic Markers of Resistance to ß-Lactams in Gramnegative Microorganisms by PCR Diagnosis].

Fifteen specimens of the hemoculture and 89 specimens of the broncho-alveolar lavage were used in the study. Monocultures of gramnegative bacteria resistant to cefotaxime, cefepime, imipenem and meropenem were isolated from the specimens. The PCR method with detection of the results in the real time regimen (PCR test-system Litekh) provided detection of the beta-lactamase genes: bla(CTX-M-like) (72/104, 69.2%), bla(NDM) (6/104, 5.8%), bla(VIM) (49/104, 47.1%) and bla(OXA48-like) (59/104, 56.7%). There was identified correlation between the phenotype of resistance of Acinetobacter spp., Pseudomonas aeruginosa, Klebsiella pneumoniae and Escherichia coli to cefotaxime and carbopenems and detection of the bla(CTX-M-like) and bla(NDM) genes. At the same time, up to 70% of the K. pneumoniae isolates from the biological specimes positive with respect to the presence of the carbapenase bla(VIM) and bla(OXA48-like) genes demonstrated their phenotypic susceptibility to carbopenems. The results of the study confirmed the prognostic value of the genetic diagnosis for improvement of the routine bacteriological investigations.