[FEATURES OF MASS-SPECTROMETRIC PROTEIN PROFILES OF STRAINS OF BRUCELLOSIS CAUSATIVE AGENT DURING PREPARATION OF CULTURE ON VARIOUS NUTRIENT MEDIA].

AIM: Carry out comparative analysis using time-of-flight mass-spectrometry with matrix laser desorption/ionization (MALDI-TOF MS) of protein profiles of brucellosis causative agents (Brucella melitensis Rev-1 and Brucella abortus 19BA), cultivated in various nutrient media: Albimi agar, brucellagar and erythrit-agar. MATERIALS AND METHODS: Vaccine,strains: Brucella melitensis Rev-1 and Brucella abortus 19BA. Protein profiling in linear mode on Microflex "Bruker Daltonics" MALDI-TOF mass-spectrometer. RESULTS: A number of characteristic features of brucella mass-spectra was detected: in particular, preservation of the total qualitative composition of protein profiles of cultures and significant differences in the intensity of separate peaks depending on the nutrient medium used. CONCLUSION: Based on the analysis of the data obtained, use of Albimi agar as the nutrient medium for preparation of brucella culture samples for mass-spectrometric analysis was shown to be optimal.

[THEORETICAL ASPECTS OF CREATION OF SPECIFIC PREPARATIONS FOR ETIOTROPIC THERAPY AND PROPHYLAXIS OF WEST NILE FEVER AND CRIMEAN HAEMORRHAGIC FEVER].

Problematic issues on creation and practical introduction of specific immune biologic prepa- rations for therapy and prophylaxis of natural-foci arbovirus infections - West Nile fever (WNF) and Crimean hemorrhagic fever (CHF), that are not available until now, are examined. Persistent natural foci of WNF and.CHF with epidemic manifestations have formed in Southern and North Caucasian Federal Districts of Russia. Markers of West Nile virus are being detected in central regions of Russia and Siberia, and the presence of fraction of population immune to this infection is also detected. Analysis, of literature sources that has been carried out forms a theoretical basis for creation ofnovel specific preparations for etiotropic, therapy and prophylaxis ofWNF and CHE. Use of blood from healthy donors with sufficiently high titers of class G immunoglobulins residing in certain subjects of Russian Federation in natural foci territories with the most intensive epidemic process is possible as raw material.

[EFFECT OF INTACT BIONANOSTRUCTURES--LIPOSOMES ON BIOCHEMICAL AND IMMUNOLOGICAL STATUS OF THE ORGANISM].

Due to features of its structure and biological properties, liposomes became not only a useful model for study of cell membranes of various organs and tissues, but also unique transporters of drugs and vaccines in the organism. At the same time, biological effect on humans and animals of the preparations included into them significantly increases. An increasingly wider use of liposomes in medicine, biology and certain adjacent specialties necessitates further studies of their effect on macroorganism, generalization, systematization and analysis of already available data. Questions of effect of intact and "empty" liposomes on biochemical and immunological parameters of the organism are examined in the review. Analysis of the ability of liposomes to render biological effect depending on their composition, preparation method and administration route is carried out. These data should be taken into consideration during creation of liposome drugs and vaccines.

[Experimental data on avian influenza virus detection using magnetic immunosorbents].

To increase the sensitivity and specificity of detecting high-pathogenic avian influenza variant (HSN1), laboratory studies were conducted at low virus concentrations in water samples, by using magnetic immunosorbent (MIS) test systems and selective avian influenza virus concentrating units. MIS-based selective virus concentrating, followed by rapid assays (ELIZA and RT-PCR), detect the low concentrations (as low as 10 nm in 5,000 ml of water) of avian influenza A/H5N1 antigen and RNA. The method developed opens new avenues for indication of an avian influenza pathogen in different environmental objects, including the water of surface water reservoirs of unlimited volume, with varying pollutions and low virus concentrations.

[Detection of the circulation of Crimean-Congo hemorrhagic fever virus in the piedmont steppes of the North Caucasus].

The laboratory verified cases of Crimean-Congo hemorrhagic fever (CCHF) in the piedmont steppes of the North Caucasus (Malgobeksky District, Republic of Ingushetia) are first described. The source of the first infection was Ixodidae ticks; three subsequent sources were contacts with the bloody discharges from patients. CCHF virus genome was detected in the blood of the cattle from an epidemic focus and in the pools of the Ixodes ticks Haemaphysalis parva Neum., 1897 and Boophilus annulatus Say, 1821, taken from cattle. The problem of including the piedmont steppes of the North Caucasus into the CCHF nosological area is discussed.

[Characterization of Bacillus anthracis typical virulent test strain 81/1].

The results of the prolonged and many-sided study of B. anthracis strain 81/1 by different authors are presented. The cultural and morphological, biochemical, antigenic, molecular-genetic characteristics of this strain give grounds for regarding it as a typical test strain to be used for the determination of the vaccines immunogenicity, the effectiveness of antibiotics and immunomodulators.

[Crimean-Congo haemorrhagic fever in southern Russia]. / Krymskaia-Kongo remorragicheskaia likhoradka na iuge Rossii.

At the end of the 20th--the beginning of the 21st century activation of a natural focus of Crimean-Congo haemorrhagic fever (CCHF) in southern Russia was noted. As a consequence, in 2002 outbreaks and sporadic cases of this disease were registered on the territory of 6 out of 13 administrative units of the Southern Federal District. To minimize the epidemiological consequences of the aggravating epidemiological situation considerable efforts and means were required from health care organs and institutions of the state sanitary and epidemiological service, including essential financial expenditures. The results of natural foci of CCHF survey, obtained by 2002, as well as main trends of prophylactic and antiepidemic interventions are presented. Scientific research and practical observations made it possible to work out a number of methodological regulations concerning the diagnostics, treatment and prophylaxis of CCHF.

[Detection of plague microbe in the fleas by polymerase chain reaction by using magnetic immunosorbents]. / Obnaruzhenie chumnogo mikroba v blokhakh metodom polimeraznoi tsepnoi reaktsii s ispol'zovaniem magnoimmunosorbentov.

The polymerase chain reaction (PCR) with and without pre-treatment of the samples of the fleas Citellophilus tesquorum with magnoimmunosorbents was used to detect the plague microbe in the samples containing 142, 168, 193 or more bacterial cells. PCR analysis reduced the time of tests of ectoparasites for plague to 6 hours. The use of a combination of PCR and magnoimmunosorbent simplified the most time-consuming and longest stage of isolation of plague microbial DNA from the suspensions of the fleas, which allows the time of a test to be reduced to 3 hours. The administration of an affine sorbent having magnetic properties during treatment of samples with plague-infected exoparasites for PCR analysis makes it possible to have a specific concentration of Yersinia pestis and to prevent the inhibitory effect of flea tissues on the polymerase chain reaction, to use the boiling method for isolating microbial DNA and for disinfecting the material to be tested.

[Emergency situations in the North Caucasus and role of specific immunoprophylaxis in eradication and prevention of epidemiological consequences]. / Chrezvichainye situatsii na Severnom Kavkaze i rol' spetsificheskoi immunoprofilaktiki v likvidatsii i preduprezhdenii ikh épidemiologicheskikh posledstvii.

Information on emergency situations (ES) on the territory of the North Caucasus from the 90s of the last century till the present moment is presented. The importance of the specific immunoprophylaxis of infections for the liquidation of epidemiological aggravations in ES and for their prevention is shown. Information on the scope of specific immunoprophylaxis in the zones of ES and on different approaches to its carrying out. The conclusion was made on the expediency of making corrections in the normative documents of the Ministry of Health of the Russian Federation, regulating planned vaccinations and vaccinations made on epidemic indications in connection with different kinds of ES.

[Basic measures for prevention of the epidemiological consequences of a natural disaster in the North Caucasus in 2002]. / Osnovnye meropriiatiia po preduprezhdeniiu épidemiologicheskikh posledstvii stikhiinogo bedstviia na Severnom Kavkaze v 2002 g.

The system of organizational and prophylactic measures, carried out with a view to prevent epidemiological aggravations in connection with the natural calamity in the Southern Federal District of the Russian Federation in 2003, is described. Proposals on the improvement of the system of preventive measures, including the monitoring of the epidemiological, sanitary and hygienic situation, personnel training, technical provision, the cooperation of different services and departments, capable of ensuring the liquidation of the consequences of emergency situations of the natural character, are made.

[Epidemiologic situation in the quarantine of natural focal infections in the Southern Federal District]. / Epidemiologichesaia obstanovka po osobo opasnym prirodno-ochagovym zabolevanniam v Iuzhnom Federal'nom Okruge.

Materials on the situation in quarantine natural focal infections (anthrax, plague, tularemia, brucellosis, leptospirosis, etc.) in the Southern Federal District are presented. Experience in the provision of the epidemic safety in the above-mentioned diseases, based on the systematic surveillance on the activity of epizootological factors on the territory of the natural foci of infections, are described. The causes of the possible aggravation of the epidemiological situation in a number of nosological forms in connection with changes in natural and socio-economic factors are given. The results of the realization of regional programs, making it possible to improve the system interaction between the administrative territories, services and departments aimed at ensuring the epidemiological safety of the region, are shown.

[Improvement of methods for indication of the causative agents of natural focal infections under conditions of emergency situations with epidemiologic consequences]. / Sovershenstvovanie metodov indikatsii vozbuditelei prirodnoochagovykh infektsii v usloviakh chrezvychainykh situatsii s épidemiologicheskimi posledstviiami.

The improvement of the laboratory diagnostics of quarantine infections (QI) is aimed at obtaining the result at a shorter time, the simplification of making serological reactions, enhancing the specificity and sensitivity of the results due to the introduction of new = generation diagnostic preparations into practical use. The wide practical use of erythrocytic and immunofluorescent diagnostic preparations has shown that they cannot completely meet all requirements simultaneously. The optimized scheme of the indication of the causative agents of QI must ensure the combined use, at different stages of the analysis, of magneto-immunosorbents, enzyme immunoassay, as well as immuno-suspension preparations for the simplification and acceleration of obtaining results with sufficient sensitivity and specificity provided.

[Epidemiological situation on anthrax in the regions of the Southern Federal District in connection with the flood in June 2002]. / Epidemiologicheskaia obstanovka po Sibirskoi iazve v regionakh Iuzhnogo Federal'nogo Okruga v sviazi s navodeniem v iiune 2002 g.

Data on the epidemiological situation in anthrax in the regions of the Southern Federal District, situated in the zone of inundation, are presented. In 2002 no epidemic complications in anthrax were noted in the affected territories. The causes contributing to the appearance of anthrax cases among humans and animals are presented. Recommendations on the improvement of anti-epidemic measures are given.

[Improvement of the method for the indication of the causative agent of anthrax]. / Sovershenstvovanie metoda indikatsii vozbuditelia Sibirskoi iazve.

A combined method for the indication of the causative agent of anthrax (Bacillus anthracis), including the preparation of the material to be tested, the exposure of the magneto-imunosorbent in the sample, cultivation in selective medium, DNA extraction with subsequent testing in the polymerase chain reaction with primers to genes cap, pag and chromosomal sequence Ba813, the registration and interpretation of results, has been developed. All determinations, including the preparation of samples, last not more than 6 hours. The indication of B. anthracis by the proposed method makes it possible not only to confirm its presence in the sample Under test, but also to evaluate its epidemic potential.

[Gene typing of Bacillus anthracis strains isolated on the territory of the countries of the Confederation of Independent States]. / Genotipirovanie shtammov Sibireiazzvennogo mikroba, izolirovannykh na territorii stran SNG.

Thirty eight B. anthracis strains isolated on the territory of the former USSR from different sources at different periods were studied by the method of the multilocus analysis of 6 chromosomal and 2 plasmid regions of B. anthracis genome with a variable number of tandem repeats. The strains belonged to 18 different genotypes; of these, 14 genotypes were described for the first time. The analysis of the genetic relationship of the strains gave grounds to suggest that on this territory both closely related strains and strains whose genotypes were remote from those peculiar to the greater part of other strains could occur. The strains belonging to subgroup A1a of molecular variability were "endemic" for the European part of the former USSR. A modification of the method of gene typing was proposed, which permitted it to be made without the use of an automatic sequencer; this made it possible to greatly widen the circle of laboratories where this method of research could be used.

[Leptospirosis in the Southern Federal District of the Russian Federation]. / Leptospiroz v Iuzhnom Federal'nom Okruge Rossiiskoi Federatsii.

The retrospective and operative epidemiological analysis of leptospirosis morbidity on the administrative territories of the Southern Federal District during the period of 1990-2002 was carried out. The dynamics and tendencies of morbidity in individual administrative units of the region were shown. Materials on the negative influence of the emergency situation arising in the region as the result of the natural calamity in summer 2002 were presented. As shown in this work, timely decisions on the stabilization of epidemiological situation taken at the period of high flood and inundation in summer 2002 in the Southern Federal District and their efficient execution made it possible to keep the epidemiological situation in leptospirosis under control.

[Current state of epidemiologic surveillance of natural foci of plague of North Caucasus]. / Sovremennoe sostoianie épidemiologicheskogo nadzora v prirodnykh ochagakh chumy Severnogo Kavkaza.

Information on the epizootic situation in plague in the natural foci of North Caucasus and on the influence of a number of anthropogenic and natural factors on this situation is presented. The data given in this work indicate that under the conditions of the anthropogenic transformation of landscapes the character of the epizootic manifestations of plague is changed and new factors, capable of aggravating epidemiological situation, appear. In addition, some other factors must be considered, such as the insufficient financing of reliable field surveys at present, the impossibility of making reliable epizootological studies due to causes of the social character (armed conflicts), thus making it impossible to evaluate, with a sufficient degree of reliability, the real epizootic state of a number of territories and, therefore, the risk of human infection. In this connection the necessity to carefully plan prophylactic measures and measures aimed at the localization and liquidation of the probable foci of infection arises.

[Use of affinity interaction of biologically active substances in practice of public health system]. / Vozmozhnosti primeneniia affinnogo vzaimodeistviia biologicheski activnykh veshchestv v praktike zdravookhraneniia.

AIM: To determine the potentialities of use of affinity interaction of immobilized biologically active substances (bacterial cells or their fragments, toxins, antigens of various chemical nature, immunoglobulins, enzymes, gangliosides, etc.) for medical practice. MATERIALS AND METHODS: Emulsion polymerization of acrylamide monomers in the gaseous nitrogen current was used as a basic method for preparation of solid-phase magnetic immunosorbents (MIC). A procedure for preparation of siliceous MIC was also applied. The prepared MICs were used a solid phase in enzyme immunoassay and immunofluorescence assay and the recorded data were compared with those of studied conventionally used in practical medicine. RESULTS: The use of MIC made it possible to detect pathogens of particularly dangerous infections in large volumes of the samples contaminated with another microflora. With the proposed MIC, one can stand a good chance of surveying large contingents of the population, of obtaining the quantitative results in shorter periods to establish a diagnosis. With this, the sensitivity and specificity of immunoassays substantially increase. Whether MIC may be used as selective hemosorbents to remove specific antibodies from the blood of patients with rheumatic diseases for therapeutic purposes was studied. CONCLUSION: The findings are indicative of wide potentialities of use of affinity interaction of biologically active substances immobilized on inert carriers with the inserted magnetic material in the laboratory diagnosis of diseases of both infectious and autoimmune nature, which may be widely used in the in- and outpatient settings.

[Affinity sorbents having magnetic properties in the clinical picture and diagnosis of communicable and non-communicable diseases]. / Affinnye sorbenty s magnitnymi svoistvami v klinike i diagnostike infektsionnykh i neinfektsionnykh zabolevanii.

Affine magnetic sorbents which have no analogs in the practice of our country have been for the first time developed for the rapid diagnosis of various life-threatening diseases (plague, cholera, anthrax, glanders, meliodosis, tularemia, leptospirosis, dysentery, viral hepatitis A) and for the identification of their causative agents. The efficacy of new magnet-controlling test systems has been repeatedly confirmed by their applications in epidemiological events and emergencies: in the epidemiological surveillance of viral hepatitis A in Stavropol and in the Caucasian Mineralnye Vody towns, Stavropol Territory (1994), in the identification of cholera patients, in the detection of transmission factors, when monitoring during large epidemic out-bursts of cholera in Stavropol (1990), Daghestan (1994), as well as in the microbiological monitoring during military conflicts in the Chechen Republic (1995). The application of the sorbents has shown that their sensitivity is 4-5 times as much as that of conventional serological assays. In addition, biotechnologies for the production of polyacrylamide and composite aluminosilicate affine immunosorbents with magnetic properties have been developed. They have been used as the basis for designing immobilized granulated antigen reagents for the immunodiagnosis, differential diagnosis, evaluation of the time course and severity of a disease, the efficiency of therapy in patients with systemic scleroderma, proliferative arthritis, systemic lupus erythematosus, juvenile rheumatoid arthritis, osteochondrosis.