Genetic Diversity of Listeria Detected in the Production Environment of Meat Processing.

The safety of food production as concerns Listeria is the key to the sanitary wellbeing of manufactured products. Molecular-genetic methods for the analysis of Listeria, including whole-genome sequencing, are effective in monitoring persistent contaminants and in the epidemic investigation of cases of foodborne infections. They have been adopted in the European Union, United States, and Canada. In Russia, multilocus and whole-genome sequencing has proven itself in the analysis of clinical food isolates and Listeria from the environment. The objective of the study was molecular-genetic characterization of Listeria detected in the industrial environment of meat processing. To characterize the Listeria isolates, microbiological methods were used according to GOST (State Standard) 32031-2012, as well as multilocus sequencing, including the analysis of seven housekeeping genes and four virulence genes, as well as whole-genome sequencing. In swabs that were positive for the presence of Listeria spp. taken at two meat-processing plants in Moscow, Listeria monocytogenes constituted 81% and L. welshimeri 19%. The predominant genotype (Sequence Type, ST) of L. monocytogenes was ST8. The variety was supplemented with ST321, ST121, and ST2330 (CC9 (Clonal Complex 9)). L. welshimeri, which prevailed in the second production, was represented by ST1050 and ST2331. The genomic characteristics of L. welshimeri isolates confirmed that they have high adaptive capabilities both as concerns production conditions (including resistance to disinfectants) and the metabolic peculiarities of the gastrointestinal tract of animals. L. monocytogenes CC9 and CC121 are also correlated with food production in other countries. However, L. monocytogenes CC8 and CC321 can cause invasive listeriosis. The concordance in the internalin profile of the ST8 isolates from the industrial environment with the clinical isolates ST8 and ST2096 (CC8) is a cause for concern. The study showed the effectiveness of molecular-genetic methods in determining the diversity of Listeria detected in the production environment of meat processing, and laid the foundation for monitoring of persistent contaminants.

Polyfunctional catalysis in conversion of light alkenes.

Light alkenes are among the main petrochemical intermediate products, the consumption of which is steadily growing. Using ethylene as an example, the possibilities of using polyfunctional heterogeneous catalysts for carrying out practically important reactions of its oligomerization, alkylation, and metathesis were considered. Particular attention was paid to catalysts for the conversion of ethylene to propylene.

[Physical therapy in the rehabilitation of patients after endoprosthetic replacement of major joints in the lower extremities: a scientometric analysis of evidence-based studies]. / Fizicheskaia terapiia v reabilitatsii patsientov posle éndoprotezirovaniia krupnykh sustavov nizhnikh konechnostei: naukometricheskii analiz dokazatel'nykh issledovanii.

BACKGROUND: In recent decades, the volume of high-tech medical care in the field of orthopedics and traumatology, including endoprosthetic replacement of major joints (MJs) (the hip joint and/or the knee joint) in the lower extremities (LE) (LEMJ), has substantially increased. In this connection, there are an increasing number of patients in need of medical rehabilitation, to solve the problems of which needs the effective physical and rehabilitation medicine (PRM) techniques proven during researches to be introduced into practice. OBJECTIVE: To analyze evidence-based studies containing sound data on the use of PRM technologies in the rehabilitation of patients after endoprosthetic replacement of LEMJs, to identify the most effective PRM technologies and to formulate recommendations for their use for practitioners, which are based on the evidence obtained during the analysis. MATERIAL AND METHODS: The paper is based on the scientometric analysis of 241 studies conducted in 2000 to 2018, which were devoted to the use of physical exercises and PRM technologies in the rehabilitation of patients after endoprosthetic replacement of LEMJs. RESULTS: Over the past decade, there has been a tangible rise in the number of studies on endoprosthetic replacement of LEMJs. Some of the most studied PRM technologies having the proven effect are physical exercises in combination with neuromuscular electrical stimulation, kinesiotherapy, cryotherapy, and pressure therapy that is effective in preventing thromboembolism after surgery. CONCLUSION: The use of PRM technologies in the rehabilitation of patients after endoprosthetic replacement of LEMJs should be based on the results of high-quality randomized controlled clinical trials, which serve as the basis for the development of clinical recommendations. The process of analyzing the data of studies should be regular.

[On the history of the problem of the rehabilitative treatment of the subjects suffering wounds in the maxillofacial region with the concomitant disorders of the speech function]. / K istorii voprosa o reabilitatsii cheliustno-litsevykh ranenykh s narusheniem rechevoi funktsii.

The authors made an attempt to highlight the issues of rehabilitation of the patients suffering wounds in the maxillofacial region with the concomitant disorders in the function of the organs of speech. The secondary objective of the study was to prove that rehabilitation of such patients is possible only by means of the joint efforts and close cooperation of dentists, physiotherapists, and speech therapists. The results of the clinical observations of the most severe cases of the impaired speech function obtained in the evacuation hospitals have been considered, with the special emphasis placed on the leading role of the speech therapist in the rehabilitation of such patients. A scheme of therapeutic physical exercises and speech therapy is presented that includes the correction and development of respiration. The guidelines are proposed for the breathing exercises and exercises for the muscles of the shoulder girdle, neck, pharynx, tongue, and the soft palate, chewing-articular muscles and mimic-articular muscles as well as for the correction of open and closed rhinolalia. Special attention is given to the implementation of socio-psychological rehabilitation for the restoration of the stable, clear and comprehensible speech function. The rehabilitation teams have been organized for the first time comprised of the maxillofacial surgeon, the orthopedic dentist, the physiotherapist, the speech therapist, and the physiotherapist. The classification of the disorders of interest have been developed based on the available data concerning the anatomical-physiological and sound-producing disturbances. The methods for the restoration of the speech function in the patients presenting with the injuries to the maxillofacial region with the concomitant disorders of the speech function are described together with the newly developed modalities of physical therapy and speech therapy.

[Clinical manifestations of Legionella pneumonia in hematology patients].

AIM: To detect the most common clinical manifestations of Legionella pneumonia (LP) in immunocompromized patients. SUBJECTS AND METHODS: Clinical manifestations, the results of investigation of bronchoalveolar lavage fluid (BALF) and urine, and the data of lung computed tomography (CT) were studied in patients with blood system diseases and acute respiratory failure (ARF). RESULTS: The diagnosis of LP was verified in 8 (10.5%) of 76 patients with blood system diseases and ARF. The disease manifested as fever, higher concentrations of inflammatory markers (procalcitonin, fibrinogen), ARF, hypoxemia, and infiltrative lung injury. Six of the 8 patients were switched to mechanical ventilation. Lung CT showed no pathognomonic signs. Five of the 8 patients were observed to have renal dysfunction. The diagnosis of LP was made on the basis of the results of BALF examination in 7 patients and urinary antigen detection in 1. The disease was caused by Legionella pneumophila serogroup 1 in 3 patients and by L. pneumophila of other serogroups in the other patients. Therapy with respiratory fluoroquinolones was performed in 5 patients. Three patients died from progressive ARF and hypoxemia. BALF results were obtained after their death and therapy for legionellosis was not initiated. CONCLUSION: The incidence of LP is 10.5% in hematology patients. The clinical manifestations of legionellosis are nonspecific; its diagnosis requires bacteriological and/or serological evidence. Due to the high risk of death, it is reasonable to preuse respiratory fluoroquinolones or macrolides in immunocompromized patients with progressive ARF and suspected Legionella pneumonia before diagnosis.

Performic acid for advanced wastewater disinfection.

The disinfection efficiency of performic acid (PFA) against various microbial contaminants has been studied in municipal secondary effluent. The study demonstrated that PFA provides rapid, efficient and safe disinfection, degrading both bacteria and viruses even at low doses. The resistance order starting from the most resistant microorganism is as follows: MS2-coliphages > DNA-coliphages > enterococci and Escherichia coli. PFA is also efficient in the elimination of Salmonella spp., Clostridium perfringens spores and Giardia cysts. The results showed that a PFA dose as low as 0.5-1 mg L(-1) with contact time of 10 min was efficient in achieving and maintaining for 72 h the disinfection level required for unrestricted agricultural water reuse (≤3 log units for faecal coliforms). However, the optimal dose will depend on the quality of wastewater. Regarding the formation of by-products during disinfection with PFA, very low amounts of hydrogen peroxide and organic per-acids were observed; active oxygen was not detected. The amounts of adsorbable organically bound halogens (AOX) compounds formed were significantly lower compared to the amounts generated during chlorine disinfection. This chlorine-free solution enables compliance with microbiological criteria for various water reuse applications and is already on the market for advanced disinfection.

[Bactericidal action of non-thermal plasma on biofilms formed in vitro and within a root channel].

AIM: Evaluation of efficiency of non-thermal plasma as bactericidal agent affecting biofilms formed in vitro and on walls of a root channel. MATERIALS AND METHODS: The multiple antibiotic resistant strain Staphylococcus epidermidis isolated from pulpitis was used. Biofilms formed in vitro on the plastic surface and ex vivo at the walls of the root canal were treated with plasma torch formed by argon:air (9:1) mixture eradiated with 100 kHz electrtomagnetic field. Bacterial viability was determined by plating and by differential Live/Dead labeling. RESULTS: The dose-dependent decrease in living bacteria was demonstrated. The three-step kinetics ofbacterial killing was observed. Total elimination ofup 10(9) CFU/sample was reached at exposition of 240 s or more. CONCLUSION: The non-thermal plasma effectively destroyed bacterial biofilms within root channels.

[The actual incidence of the population in the RF subject: assessment of economic effect (losses)].

In the paper there is presented an analysis and evaluation of the economic losses associated with the actual incidence of the population of the Altai Region on disease classes "Poisoning by drugs, medicaments and biological substances" (T36-T50) and "Toxic effect of substances, mainly non-medical purpose" (T51-T65), including the assessment of the underproduced product in economy of the region in monetary terms, assessment changes in cash flows on the budgets of the Russian Federation (tax receipts). The time period of analysis on disease classes is 5 years (2007-2011).

[Features of legionella biofilm formation in artificial and natural water systems].

AIM: Study the ability to form monospecies and associative biofilms as a characteristic oflegionella strains and features of organization of natural legionella biofilms in potentially dangerous water systems. MATERIALS AND METHODS: Comparative evaluation ofthe ability of 28 strains of Legionella spp. to form biofilms was determined in water according to previously developed procedure. Samples from biofilm of industrial enterprise coolers and systems of hot water supply of public buildings (hotels, trade centers, hospitals) were studied. Biofilms were studied by scanning and transmission electron microscopy methods. RESULTS: Legionella strains are divided into 3 groups by the ability to form biofilms. L. pneumophila BLR-05 strain that has the most pronounced ability to form monospecies biofilm and persistence in association with Pseudomonas aeruginosa was detected. Formation of massive legionella biofilm in association with bacteria of other taxonomic groups was detected on protective antibacterial filters in the system of hot water supply of a department of a therapeutic prophylaxis institution in the course of 2-3 weeks. Legionella biofilms on the surface of coolers resemble an aggregate of fungi, bacteria and blue-green algae enclosed into matrix. CONCLUSION: The ability to form artificial monospecies and associative biofilm may be a useful characteristic of legionella strains for evaluation of their adhesion and be used to evaluate epidemiological significance of the isolated strains. Prevention of formation of natural legionella biofilms in potentially dangerous water systems is necessary as an essential component of modern strategy of legionellosis prophylaxis.

[Prevalence of glucosyl transferase Lgt among Legionella pneumophila strains isolated from various sources].

AIM: Determine various members of Lgt glucosyl transferase family among microorganisms of Legionellaceae genus from museum collection and legionellae strains recently isolated in the Russian Federation and Germany. MATERIALS AND METHODS: Presence of 3 types of glucosyl transferase were determined in 73 strains of L. pneumophila and Legionella spp. Glucosyl transferase activity of 3 types (Lgt1, Lgt2 and Lgt3) was determined by western blotting and PCR method. RESULTS: Lgt1 and Lgt3 were detected only in members of L. pneumophila independently of isolation source and were absent in Legionella spp. strains. Lgt2 is absent in Legionella spp. strains and is detected in not all the L. pneumophila strains. Comparative analysis of detection frequency of Lgt2 in clinical strains and L. pneumophila strains isolated from the environment showed that the protein is detected in clinical strains more frequently (46%) compared with strains from the environment (23%). CONCLUSION: Lgt1 and Lgt3 as species specific markers could be used for practical purposes for identification of L. pneumophila strains. High frequency of Lgt2 isolation in clinical strains of L. pneumophila isolated from lung tissue in lethal cases of legionellosis compared with strains isolated from the environment requires a more detailed study of functional activity and substrate specificity of the glucosyl transferase.

[Serologic characteristic of Legionella pneumophila strains isolated from potentially dangerous water systems in Russian Federation in 2007 - 2011].

AIM: Study serologic diversity of Legionella pneumophila strains circulating in potentially dangerous water systems in Russian Federation by using an international panel of monoclonal antibodies. MATERIALS AND METHODS: Serotyping of 234 L. pneumophila strains isolated from coolers of industrial facilities and systems of hot water supply in Russian Federation in 2007 - 2011 was performed by enzyme immunoassay by using an international panel of monoclonal antibodies. RESULTS: Membership of the isolated strains in 14 L. pneumophila serogroups and in 7 subgroups of serogroup 1 was established. Among the isolated cultures serogroup 1 and 6, and Oxford and Heysham subgroup strains were predominant. L. pneumophila serogroup 1 strains were predominant in cooler water, and serogroup 6--in the hot water supply systems. 7 L. pneumophila strains of the serogroup 1 were typed by monoclone MAb 3/1 associated with LPS epitope that is characteristic for the most epidemically significant legionella strains. CONCLUSION: Typing by using international panel of monoclonal antibodies for characteristic and evaluation of epidemical significance of legionella strains being isolated form potentially dangerous water systems is the most informative and methodically accessible to a wide range of biological laboratories.

[A case of legionellesis pneumonia verified by isolation of Legionella pneumophila serogroup 1 from bronchoalveolar lavage fluid treated with levofloxacine and tigecycline].

A male patient received non-chemotherapeutic drugs which induced deep neutropenia complicated with sepsis, bilateral pneumonia, acute respiratory insufficiency. Artificial pulmonary ventilation was applied. The examination of bronchoalveolar lavage showed the presence of the culture L. pneumophila (serogroup 1) in a concentration 2 x 10(3) CFU/ml. Antibacterial therapy with levofloxacin in a dose 1000 mg/day was conducted. In a week not only L.pneumophila but also Acinetobacter baumanii was isolated from bronchoalveolar lavage. Tigecyclin was added to levofloxacin treatment. Two air cavities were found in the left lung. The treatment reduced the size of these cavities, infiltrative changes in the lungs and respiratory insufficiency regressed. The patient was discharged from hospital This case is the first case in Russia of L.pneumophila isolation from bronchoalveolar lavage. The case is also characterized by use of tigecycline for treatment of combined legionella and akinetobacterial infection and cavities in the lungs in legionella pneumonia.

[Comparative analysis of therapeutic efficiency of modern methods for remodeling skin scars: a milticenter randomized study].

The authors describe and substantiate therapeutic effects of physical factors on skin scars in accordance with the basic principles of evidence-based medicine. These factors are shown to significantly change the structure and metabolism of the cicatrical issue by virtue of their collagen-modulating and anti-fibrotic action. The results of application of various physiotherapeutic modalities suggest high efficacy of their combination for the correction of different types of skinscars.

Mutator effects and mutation signatures of editing deaminases produced in bacteria and yeast.

Enzymatic deamination of bases in DNA or RNA leads to an alteration of flow of genetic information. Adenosine deaminases edit RNA (ADARs, TADs). Specialized cytidine deaminases are involved in RNA/DNA editing in lipid metabolism (APOBEC1) and in innate (APOBEC3 family) and humoral (AID) immunity. APOBEC2 is required for proper muscle development and, along with AID, was implicated in demethylation of DNA. The functions of APOBEC4, APOBEC5, and other deaminases recently discovered by bioinformatics approaches are unknown. What is the basis for the diverse biological functions of enzymes with similar enzyme structure and the same principal enzymatic reaction? AID, APOBEC1, lamprey CDA1, and APOBEC3G enzymes cause uracil DNA glycosylase-dependent induction of mutations when overproduced ectopically in bacteria or yeast. APOBEC2, on the contrary, is nonmutagenic. We studied the effects of the expression of various deaminases in yeast and bacteria. The mutagenic specificities of four deaminases, hAID, rAPOBEC1, hAPOBEC3G, and lamprey CDA1, are strikingly different. This suggests the existence of an intrinsic component of deaminase targeting. The expression of yeast CDD1 and TAD2/TAD3, human APOBEC4, Xanthomonas oryzae APOBEC5, and deaminase encoded by Micromonas sp. gene MICPUN_56782 was nonmutagenic. A lack of a mutagenic effect for Cdd1 is expected because the enzyme functions in the salvage of pyrimidine nucleotides, and it is evolutionarily distant from RNA/DNA editing enzymes. The reason for inactivity of deaminases grouped with APOBEC2 is not obvious from their structures. This can not be explained by protein insolubility and peculiarities of cellular distribution and requires further investigation.

Calorimetric and structural studies of tetrabutylammonium chloride ionic clathrate hydrates.

Three ionic clathrate hydrates of different water content are formed in the binary system tetrabutylammonium chloride-water. The data on heats of fusion of these hydrates measured by DSC are presented. Single-crystal and powder X-ray diffraction studies of ionic clathrate hydrates of tetrabutylammonium chloride have been carried out. Structural justification of the occurrence of three different ionic clathrate hydrates derived from tetragonal structure-I in the (C(4)H(9))(4)NCl-H(2)O system has been found for the first time. A novel mode of hydrophilic inclusion of the halide anion with displacement of two hydrogen-bonded host water molecules has been revealed. Structural data obtained are indicative of the possibility of location of tetrabutylammonium cations in combined cavities formed with participation of D-voids.

[Rate and level of contamination of potentially dangerous water supply facilities in Moscow region by Legionella pneumophila].

AIM: To assess rate and level of contamination by Legionella pneumophila of cooling water systems in industrial facilities as well as hot water supply systems of administrative buildings in Moscow region. MATERIALS AND METHODS: Cooling water systems of 8 industrial facilities and hot water supply systems of 12 administrative buildings or complexes located in Moscow or Moscow region were examined. Samples of water, washes and biofilms were studied by bacteriologic methods and RT-PCR. Results. Significant level of contamination of water systems by L. pneumophila was revealed in examined objects. Rate of contamination of cooling water systems in industrial facilities was 70%. The agent was detected in stagnant, end-capped, and rarely used segments of all hot water supply systems during decrease of water temperature to 36-52 degrees C. Visual detection of natural biofilms on the object correlated with high concentration of L. pneumophila in water samples. In some cases, associations of L. pneumophila with Pseudomonas aeruginosa were detected, including water samples from supply systems of 2 healthcare facilities. CONCLUSION: Obtained results confirm the importance of implementation of modem concept of legionellosis prevention in our country, based on regular quantitative monitoring for Legionella in potentially dangerous water objects and conduction of preventative measures if contamination exceeds acceptable level.

Enhanced activity of the CREB co-activator Crtc1 in LKB1 null lung cancer.

Activation of Crtc1 (also known as Mect1/Torc1) by a t(11;19) chromosomal rearrangement underlies the etiology of malignant salivary gland tumors. As LKB1 is a target for mutational inactivation in lung cancer and was recently shown to regulate hepatic Crtc2/CREB transcriptional activity in mice, we now present evidence suggesting disruption of an LKB1/Crtc pathway in cancer. Although Crtc1 is preferentially expressed in adult brain tissues, we observed elevated levels of steady-state Crtc1 in thoracic tumors. In addition, we show that somatic loss of LKB1 is associated with underphosphorylation of endogenous Crtc1, enhanced Crtc1 nuclear localization and enhanced expression of the Crtc prototypic target gene, NR4A2/Nurr1. Inhibition of NR4A2 was associated with growth suppression of LKB1 null tumors, but showed little effect on LKB1-wildtype cells. These data strengthen the role of dysregulated Crtc as a bona fide cancer gene, present a new element to the complex LKB1 tumorigenic axis, and suggest that Crtc genes may be aberrantly activated in a wider range of common adult malignancies.

[Using real-time polymerase chain reaction for detection of Legionella in objects of the environment].

AIM: To assess efficacy of using the method of quantitative detection of Legionella in objects of the environment by real-time polymerase chain reaction (RT-PCR). MATERIALS AND METHODS: For the development of the assay, genus-specific primers from gene coding 16S rRNAas well as species-specific primers for detection of Legionella pneumophila on the basis of mip gene sequence. For quantitative detection of L. pneumophila calibration samples of pGEM plasmid containing fragment of the mip gene in known concentration were used. Samples of water and biofilms obtained from cooling stacks of production plants, systems of autonomic water supply, humidification blocks of centralized systems of air conditioning were studied. RESULTS: Correlation of results obtained with RT-PCR and bacteriologic methods was shown during monitoring of potentially dangerous water objects as well as during epidemic outbreak of Legionella infection. Importance of samples preparation stage, during which considerable losses of DNA and inhibition of reaction could occur, is underlined. Disinfection measures on the studied objects significantly influenced on the results of the RT-PCR and can lead to false positive results. CONCLUSION: Obtained results confirm usefulness of testing of potentially dangerous water objects on the presence of Legionella based on the preliminary screening with RT-PCR for the 24 hours followed by bacteriologic testing of samples for 8 - 12 days.

[Analysis of prevalence and variability of Legionella pneumophila and Legionella spp. strains on the basis of study of allelic profiles].

AIM: To analyze prevalence and variability of Legionella strains isolated in town Verkhnaya Pyshma located in Sverdlovsk region during prophylactic surveillance of potentially dangerous water objects in 2007 - 2008. MATERIALS AND METHODS: Sequencing of mip gene was conducted for identification of species of Legionella. Multi-locus sequence typing was used for describing of allelic profiles of Legionella pneumophila strains. RESULTS: Five firstly identified on Russian territory strains of Legionella species were deposited in institute's collection. Sixty-three strains of L. pneumophila belonging to 28 sequence types were characterized. Relation between strains isolated in industrial building and from water supply system was demonstrated. CONCLUSION: Observations made on the basis of study of L. pneumophila strains isolated from cooling stacks of industrial plants confirmed potential danger of these objects as a source of dissemination of Legionella infection.

[Modern views on the epidemiology of legionellosis: operations procedure during epidemic outbreaks and preventive monitoring].

Key aspects of legionellosis epidemiology determining the rising role of Legionella in human infectious diseases in conditions of postindustrial community were discussed. It was shown that formation of Legionella biofilms on potentially dangerous water objects with possible further aerosol or aspiration transmission of microorganisms leads to major epidemic outbreaks and sporadic cases of infection. Main principles of actions during preventive monitoring of Legionella in the environment and epidemic outbreaks of legionellosis were described.