[Thermosensitization of tumor cells with inhibitors of chaperone activity and expression].

Effects of inhibitors of the heat shock protein 90 (HSP90) chaperone activity and inhibitors of the heat shock protein (HSP) expression on sensitivity of HeLa tumor cells to hyperthermia were studied. It was found that nanomolar concentrations of inhibitors of the HSP90 activity (17AAG or radicicol) slowed down chaperone-dependent reactivation of a thermo-labile reporter (luciferase) in heat-stressed HeLa cells and slightly enhanced their death following incubation for 60 min at 43 degrees C. Herein, the inhibitors of HSP90 activity stimulated de novo induction of additional chaperones (HSP70 and HSP27) that significantly increased the intracellular HSP levels. If the cells were treated with 17AAG or radicicol along with an inhibitor of the HSP induction (e.g. quercetin or triptolid, or NZ28), this fully prevented the increase in intracellular chaperone levels resulting from the inhibition of HSP90 activity and subsequent heating. Importantly, in the case of conjunction of all the three treatments (an inhibitor of the HSP90 activity + an inhibitor of the HSP induction + 43 degrees C for 60 min), the reporter reactivation was retarded yet stronger while the cell death was sharply (2-3-fold) enhanced. Such an enhancement of the cytotoxicity appears to occur owing to the "chaperone deficiency" when prior to heat stress both the functional activity of constitutive HSP90 and the expression of additional (inducible) chaperones are blocked in the cells.

[Inhibitors of the heat shock protein 90 activity: a novel class of tumor radiosensitizers].

The 90 kDa heat shock protein (HSP90) is one of major chaperones of eukaryotes which catalyzes maturation and activation of its client proteins. Among the identified client proteins there are oncogene products, hormone or growth factor receptors and key components of signaling pathways responsible for the malignant growth of tumors or their resistance to chemotherapy and radiotherapy. In the case of inhibition of the HSP90 chaperone function, such proteins are inactivated and degraded soon that leads to simultaneous blocking several pathways essential for proliferation and survival of malignant cells; therefore, pharmacological inhibitors of the HSP90 chaperone activity could be used in anticancer therapy. At present, several HSP90 inhibitors are in preclinical testing or I-III Phase clinical trials as mono-agents or in combinations with other anticancer drugs or radiation. In the present review, all the data are summarized which characterize HSP90 inhibitors as effective radiosensitizers of tumor cells. Molecular mechanisms and selectivity of the radiosensitizing action of HSP90 inhibitors are here discussed as well as a possibility of their application to improve the outcome of radiotherapy.

[Hemostatic disorders and their implication in the pathogenesis of Astrakhan rickettsial fever].

AIM: to define a role of hemostatic disorders in the pathogenesis of Astrakhan rickettsial fever (ARL). SUBJECTS AND METHODS: Platelet functional activity and plasma hemostatic parameters were studied in 89 patients of moderate ARL. RESULTS: The clinical manifestations of hemostatic disorders at the height of ARL were characterized by the appearance of typhoid maculopapular rashes in 91.4% of the patients, solitary elements of which were transformed to petechiae in 20% of cases. At convalescence (on day 10.2 +/- 1.3 of the disease), all eruptions regressed via pigmentation. At the peak of the fever, there were nasal hemorrhages and bleedings from the sites of injections; lowered platelet aggregation was detectable in the presence of thrombocytopenia at the height. Coagulation hemostasis changes were characterized only by elevated fibrinogen levels. Increased platelet functional activity and decreased fibrinogen concentrations were observed at convalescence. CONCLUSION: The basis of the clinical manifestations of ARL is hemostatic disorders due to thrombocytopenia and diminished platelet functional activity. In early convalescence, there was improved platelet aggregatability; however, the increasing trend for the rate of aggregation and the radius of aggregates suggests a risk of thrombogenesis in convalescents, which requires their follow-up with obligatory hemostatic monitoring.

[Characteristics of non invasive ultrasound determination of mechanical lens hardness].

Sixty-two patients with varying maturity cataract were examined to assess a new noninvasive method for determining the mechanical hardness of the nucleus of the lens. There was an inverse relation of the hardness of the nucleus to its relative acoustic density (delta). Comparison of the ultrasound density of the lens and the mechanical characteristics of its nucleus has provided the following classification: the lens hardness is high at delta of 0 to 0.2; average at 0.3 to 0.5, and low at 0.6 or higher. Determination of relative acoustic density by measuring the parameter delta substantially enhances the accuracy ofpreoperative prediction of the hardness of the nucleus of the lens when a surgical option for cataract removal is chosen.

[Lytic activity of aerobic sporulating bacteria]. / Liticheskaia aktivnost' aérobnykh sporoobrazuiushchikh bakterii.

The paper deals with lytic activity of 226 strains belonging to 13 species of Bacillus genus, isolated from various econiches, in respect of the cells of Staphylococcus aureus, Escherichia coli and Candida albicans as representatives of Gram-positive, Gram-negative and yeast groups of microorganisms. All the studied strains of bacilli displayed lytic activity of various extent and various spectrum of substrate specificity. Possible dependence of the extent of activity and substrate specificity of bacilli strains on their species belonging and isolation sources did not display distinctly. Two different factors of aggression and antagonism of aerobic bacilli--the capacity to lyse as well as to inhibit growth of test-cultures do not correlate and can either coincide or not coincide. The Bacillus genus and its species are characterized, to a certain extent, as a lytically active group and are estimated as the promising source of obtaining highly active producers of lytic enzymes different as to their properties. Highly active producers of lytic enzymes with the primary degree of lysis from 30% to 60% of the cells of test cultures during 1 hour of the reaction have been obtained.

[Use of PCR and nucleotide sequence analysis of the VP7 gene for detection of bovine rotaviruses and identification of their G serotypes]. / Ispol'zovatie PTSR i sravnitel'nogo analiza nukleotidnoi posledovatel'nosti gena VP7 dlia vyiavlenia rotavirusov krupnogo rogatogo skota i opredeleniia ikh G-serotipov.

A Vp7 gene fragment PCR protocol was developed to detect the bovine rotaviruses and to identify their G serotypes. The most widespread bovine rotaviruses of G serotypes (G6, G8 and G10) can be distinguished on the basis of the PCR fragment size, while other G serotypes can be differentiated through a comparative analysis of the VP7 gene fragment nuclcotide sequence. Twenty-four bovine rotavirus field isolates were detected, and their G stereotypes were determined by using the method in question. Fourteen isolates were shown to be of G6 serotype; four of them were of G8, five--of G10, and one isolate was of G11 serotype. A possibility of detecting more than one isolate by this method was shown. Finally, a feasibility of using the method for searching for rotaviruses of new bovine rotavirus G serotypes and for rotaviruses, which do not belong to a so far described G serotypes, is discussed.

[Effect of microelements on accumulation of biomass and exopolysaccharides in Bacillus subtilis strains]. / Vliianie mikroélementov na nakoplenie biomassy i ékzopolisakharidov shtammami Bacillus subtilis.

Different trace elements in the composition of nine synthesized complex compounds of N-oxides of pyridine derivatives have been studied. It has been shown that the effect of trace elements included in the composition of nutrient media on accumulation of cells and exopolysaccharides (EPS) by the strains of B. subtilis is sufficiently expressed; it is also diverse and depends on the element nature, concentration as well as on peculiarities of the bacillus cultures. The studied trace elements are separated into the conditional groups: the elements intensifying the above processes (manganese and lithium); elements repressing the processes (copper and cadmium); those retarding the accumulation of biomass but activating the secretion of EPS (nickel, boron, cobalt, zinc, iron). Concentrations of trace elements optimal for the studied productivity indices of strains have been established. Peculiarities of the trace element effect in paired and ternary combinations on the growth and biosynthesis of EPS during cultivation of strains in liquid media have been revealed.

[The effect of the nutritional sources on the synthesis of exopolysaccharides and amino acids by Bacillus subtilis strains]. / Vliianie istochnikov pitaniia na sintez ékzopolisakharidov i aminokislot shtammami Bacillus subtilis.

Dynamics of cell biomass accumulation and secretion to the medium extracellular polysaccharides and amino acids has been studied in Bacillus subtilis cultures No No 39 and 51 used to produce healing biopreparations--probiotics. The investigation data indicate to certain relation between these processes. EPS secretion in the studied cultures proceeded parallel with their growth and started in the logarithmic phase. Maximum EPS yield was observed by the beginning of the stationary phase after 10-12 h of growth. A successible change in the amino acid content in the medium was observed in the growth process of the studied bacteria: the bacteria first consumed amino acids of the initial medium and then excreted amino acids synthesized into the medium. Under the active production of EPS the content of extracellular amino acids in the medium was inconsiderable. The content of EPS was lower during accumulations of high concentrations of extracellular amino acids. Role of the medium components in regulation of the studied processes has been shown. The ratio C:N in the medium was of essential significance. The C:N ratio 2.0-3.0:1.0 was optimal both for the growth and secretion of EPS by the studied cultures while that of 1.0-1.5:1.0 was optimal for production of the extracellular amino acids. The increase of C:N ratio resulted in the decrease of metabolites secretion by the cultures.

[The effect of the acidity of the medium and of the temperature on the growth and polysaccharide excretion of Bacillus subtilis under submerged cultivation]. / Vliianie kislotnosti sredy i temperatury na rost i ékskretsiiu polisakharidov Bacillus subtilis pri glubinnom kul'tivirovanii.

Growth of Bacillus subtilis strains and excretion of polysaccharides have been studied under different environmental condition. These strains were studied as active antagonists of pathogenic microflora and were used for development of the biopreparation (probiotic), being efficient in treatment and prophylaxis of a number of bacterial infections of farm animals; the drug soon will be put into series production. The indices of accumulation of biomass of viable cells and exopolysaccharides (EPS) were used to study productivity of the strains. A possibility to increase yield in the production process was shown using the temperature factor and optimal pH of the culture medium under batch cultivation. The processes of the biomass and EPS accumulation were noncompetitive, almost coincided in time and were independent of the initial level of the pH of medium.

[Stimulation of growth and spore formation of Bacillus subtilis by optimization of carbohydrate nutrition during submerged cultivation]. / Stimulatsiia rosta i sporoobrazovaniia Bacillus Subtilis s optimizatsiei uglevodnogo pitaniia pri glubinnom kyl'tivirovanii.

A wide range of carbohydrate sources for the growth of industrial Bacillus subtilis strains were studied. The growth was mainly accelerated 2.7 and 2.5-fold by adding green syrup and glucose in a combination with xylose, respectively, in comparison to the traditional glucose. The revealed nutritional demands of these bacteria allowed us to select an optimal composition of the medium using a method of experimental design. Conditions essential for the maximum production of cells and spores were determined, and certain similarities and strain-specific features of this process were found for the strains studied. Mathematical models describing growth and spore formation during submerged cultivation of Bacillus subtilis were also proposed in this work. The antagonistic activity possessed by the strains studied was shown to be unchanged through batch cultivation. We demonstrated the possibility of direct regulation of Bacillus subtilis growth and spore formation by maintaining the concentration of carbon and nitrogen sources and other components of growth media at a certain level.

[The effect of live cultures of Bacillus subtilis on body nonspecific resistance]. / Vliianie zhivykh kul'tur Bacillus subtilis na nespetsificheskuiu rezistentnost' organizma.

Alive cultures of Bacillus subtilis, antagonists of pyogenic microflora, have been studied for their effect on functional activity of macrophagal cells and induction of endogenic serum alpha-interferon in the female mice of the SBA line. It is established that even a single intravaginal and intraperitoneal introduction of bacilli in a dose of 1 milliard cells in 0.1 ml of cultural fluid stimulates migration, absorption and especially bactericidal activity of macrophages of peritoneal exudate. Introduction of alive cultures of aerobic bacilli essentially stimulates the in vivo production of the serum and alpha-interferon induced in vitro by the Newcastle disease virus. The highest immune-stimulating effect under a single introduction of B. subtilis is achieved 66 h later and has a tendency to gradual decrease.

[The role of amino acids in intensification of Bacillus subtilis exopolysaccharide biosynthesis in deep growth conditions]. / Rol' aminokislot v intensifikatsii biosinteza ékzopolisakharidov Bacillus subtilis v glubinnykh usloviiakh rosta.

The influence of various combinations of amino acids on the Bacillus subtilis growth activity and exopolysaccharide biosynthesis has been studied. The effects of amino acid additions on these two processes have appeared to be different. The strains reveal the specificity in amino acids requirements while cultivation in liquid media. To judge by the average indirect contributions, the growth as well as the exopolysaccharide biosynthesis of B. subtilis strain 39 have been significantly affected by introduction of alanine, valine, phenylalanine, tryptophane and glycine in triple combinations into the culture medium. The exopolysaccharide production and growth of B. subtilis strain 51 have been influenced markedly by methionine, leucine, isoleucine, cystine, glycine, tryptophane and alanine. Addition of different combinations of these amino acids makes it possible to obtain the 20.2-56.3% higher biomass amounts with the simultaneous 26.8-89.6% increase of exopolysaccharide yield.

[The growth and sporulation of Bacillus subtilis under different aeration conditions]. / Rost i sporoobrazovanie Bacillus subtilis v razlichnykh usloviiakh aératsii.

The dynamics of accumulation of Bacillus subtilis 39 cells and spores under different conditions of aeration has been studied. It is shown possible to intensify the process of growth, biomass accumulation and sporulation of bacteria using aeration conditions. The growth of cells of the studied bacilli proceeds under conditions of providing the culture with oxygen according to its real demands for the whole period of fermentation. The mathematical models have been obtained which connect the aeration conditions with the rate of biomass accumulation and sporulation of the studied bacteria. It has been stated that with an increase of aeration the yield of microbic cells, degree of using the substrate by them as well as the specific growth rate become higher. Intensive aeration 4.4 = 5.8 g O2/l.h promotes the creation of the most optimal conditions for the growth of bacteria. While deteriorating the aeration to 3.2 g O2/l.h and over, the specific growth rate and yield of viable cells decrease. The optimal conditions of air feed and medium agitation rate decrease the duration of fermentation exerting no effect on the final yield of biomass. It is shown that yield of biomass and spores depends on the growth phase.

[The antagonistic action of aerobic sporulating bacteria on the causative agents of endometritis in cattle]. / Antagonisticheskoe deistvie aérobnykh sporoobrazuiushchikh bakterii na vozbuditelei éndometritov krupnogo rogatogo skota.

247 strains of 14 species of bacteria (genus Bacillus) have been studied for the antagonistic properties relative to a wide range of the cattle endometritis agents which are both museum and isolated by authors under the above pathology. Many species of genus Bacillus showed a pronounced antagonism. Active antagonists can occur in different species. Bacillus subtilis strains are the most active. Multiple antibiotic resistance of endometritis agents has no effect on the antagonistic activity of the spore-forming aerobe strains. It is shown that practically the whole range of the possible bacterial endometritis agents is sensitive to the antagonistic effect of the most active strains of aerobic bacilli. The antagonistic strains promising for further studies have been obtained.

[Extracellular amino acids of aerobic spore-forming bacteria]. / Vnekletochnye aminokisloty aérobnykh sporoobrazuiushchikh bakterii.

An ability to synthesize and accumulate in the growth medium for extracellular amino acids of 108 strains of 13 aerobic sporeforming bacteria species at deep cultivation on the simple synthetic glucose-mineral liquid nutrient medium optimized for the representatives of this genus. Has been studied the bacillus strains studied in the present conditions synthesize 19 amino acids and 2 amino carbons in different quantities and combinations. Amino acids being synthesized the most and least by these microorganisms have been determined in quantitative expression. Extracellular amino acid accumulation was proved to be a strain property not of genus. In general, the genus studied was evaluated as active one in amino acid production. The most active strains may present an interest as initials for further selective obtaining of producers of separate amino acids. The outlook for aerobic bacilli strains having marked amino-synthesizing properties and complex of other biological activities is being discussed for construction of medicinal and prophylactic of live microbial culture preparations.

[The characteristics of the microflora isolated in bovine endometritis]. / Kharakteristika mikroflory, vydelennoi pri éndometritakh krupnogo rogatogo skota.

117 pure bacterial cultures of numerous species isolated by the authors at acute postnatal pyo-catarrhal cattle endometritis have been studied for their virulence and resistance to 18 antibiotics with the aim to determine their etiologic role in the development of the given pathology and to make a prediction estimation of possible therapeutic and preventive efficiency of the antibiotics. The experiments on white mice have revealed pathogenic properties and virulence in most studied strains of bacteria of different species. The studied microflora is, mainly, resistant practically to all 18 tested antibiotics. A direct dependence is established between virulence of the microflora, isolated in case of endometritis, and its resistance to antibiotics.

[Orthopedic devices for footwear in flatness of the anterior part of the foot]. / Ortopedicheskie prisposobleniia dlia obuvi pri rasplastannosti perednego otdela stopy.

The work is dedicated to the problem of orthopedic supply of patients with flatness of the anterior part of foot. There is proposed a number of footwear orthopedic devices made of foam rubber. There is described the technology of their production with application of modern polymeric materials. There has been studied prosthetics of 50 patients with flatness of foot and revealed the positive result of application of footwear orthopedic devices made of foam rubber.

[The selection of the composition of the medium for optimizing the amine-synthetic activity of aerobic bacilli]. / Podbor sostava sredy dlia optimizatsii aminosinteticheskoi aktivnosti aérobnykh batsill.

Nutrient medium chosen as a basic one after preliminary test of several media known from literature has been optimized to intensify biosynthesis and amine nitrogen production by three strains of aerobic sporulating bacteria to culture liquid. The method of mathematical planning used in the experiments has permitted obtaining the components ratio for the medium on which production of amine nitrogen to the environment increased 2.3-3.2 times. The best variants of the optimized medium promoted an increase of the aminosynthetic activity of the studied bacteria by more than 320%. The obtained nutrient medium is appropriate for a wide screening of aerobic bacilli for their ability to synthesize amino compounds.

[The microflora in suppurative-catarrhal endometritis in cattle]. / Mikroflora pri gnoino-kataral'nykh éndometritakh krupnogo rogatogo skota.

Microflora isolated from cattle with acute postnatal pus-catarrhal endometritis has been studied. It is shown that the given pathology is caused by associations of different bacterial species. 284 strains isolated from the uterus content of 63 sick animals have been identified, their properties being studied. Pathogenic gram-negative bacilli and gram-positive pus-producing cocci are responsible for the studied pathology. Considerable inoculation capacity has been established for conditionally pathogenic microorganisms of different species and genera whose role in pathogenesis is to be found out.