PREPARATION OF MONOSPECIFIC BRUCELLA ANTI-ABORTUS SERUM BASED ON THERMOEXTRACT FROM S-FORM BRUCELLA

Objective of the study is to prepare rabbit monospecific anti-abortus serum to a Brucella S-form thermoextract, as well as to evaluate its activity and specificity. Materials and methods. Utilized was polyvalent hyperimmune serum obtained to Brucella thermoextract. It was adsorbed by a strain of heterologous species Brucella melitensis 16 N. Reference, vaccinal and field Brucella strains were used to estimate the activity and specificity of the serum produced. Results and conclusions. This serum possesses high activity (1:160, 1:320 titer) and specificity, and the results completely coincide with the data specified in the table of differential properties presented in МR 3.1.7.1189-03 “Prophylaxis and laboratory diagnostics of human brucellosis». The outlined method for manufacturing monospecific Brucella anti-abortus serum permits for a considerable decrease in biological hazard for personnel involved in the technological operations

PECULIARITIES OF LABORATORY DIAGNOSTICS OF EXPERIMENTAL BRUCELLOSIS CAUSED BY SAND L-FORMS OF CAUSATIVE AGENT

Comparative evaluation, of bacteriological, serological and PCR diagnostic methods was resulted, at experimental brucellosis in guinea pigs infected, with. S- and. L-forms of the causative agent in different periods of the infection. It was shown that Brucella S-forms were isolated, during six months, L-forms - about one month, by a bacteriological method. Positive PCR results were observed about six months after S- and. L-form infection in animals. Positive slide agglutination, of guinea pig blood sera was revealed, till 12 months by a corpuscular diagnosticum with. S- and. L-forms of Brucella. Complex use of bacteriological, PCR and. serological methods for laboratory diagnostics of atypical brucellosis caused, by Brucella L-form allows to obtain more reliable information, about epizootological-epidemiological situation in non-manifested (non-clinical) infection foci

Improvement of a test-system for the botulinus toxin screening in dot-immunoanalysis

We constructed a test system for dot-immunoassay (DIA) to accelerate definition and identification of botulinus toxins and also to refuse from application of laboratory animals for routine screening of clinical samples, foodstuff and environments. This system permitted to detect botulinus toxin during approximately 2 h in the tested samples. Sensitivity of this DIA in some cases exceeded the mice biotest. This improved method has minimum reaction to nonspecific exposures from the investigated biological substrata. It is simple to conduct. It is high efficient and expressive, does not require to use expensive equipment and the reactants, special training for the personnel. Lyophilization conditions for the immune reagents used for the test system preparation for botulinus toxin dot-immunoassay were selected. High sensitivity, specificity of the analysis are remained, stability of the preparations (periods of storage) is increased. This method is convenient to use in field conditions at extreme situations, in particular, in mobile autolaboratories for epidemiological survey

Construction of nutrient media for L-form brucella

Dry nutrient media on the basis of hepatic infusion and Siberian roach hydrolyzate are constructed. Experimental data demonstrate that these media possess high sensitivity, completely inhibit growth of S-form Brucella, do not require рН adjustment, filtration and autoclaving. They can be used for isolation, cultivation and accumulation of L-form Brucella for bacteriological diagnostics of brucellosis. As these media are accessible for transportation, they are applicable for stationary and field conditions

Using dot-immunoassay in decoding the outbreak of pseudotuberculosis in the Tomsk region

Background. Pseudotuberculosis remains a serious healthcare problem, which determines the expediency of developing the express methods for its early diagnosis. To detect the pathogen, we designed test system for dot-immunoassay (DIA) based on antibodies labeled with silver nanoparticles (SNPs) isolated from hyperimmune rabbit serum obtained against killed cells of  Yersinia pseudotuberculosis of O:1b serovariant.The aim. To assess the possibility of using dot-immunoassay for express identification of Y. pseudotuberculosis cultures isolated from clinical material and environmental objects at the initial stage of bacteriological study during laboratory diagnosis of the disease.Methods. We used the materials from the outbreak of pseudotuberculosis in the Krylovskaya Boarding School of the Bakcharsky district of the Tomsk region in 2021. Specific antibodies from hyperimmune rabbit sera obtained against Y. pseudotuberculosis 3704 particulate antigen of O:1b serotype were labeled with SNPs and used in DIA on nitrocellulose membranes with visualization of reaction results with a solution of a physical developer. The presence of the causative agent of pseudotuberculosis in the test material was inferred by the formation of gray spots of different intensity (from 4+ to 1+).Results. All Y.  pseudotuberculosis strains isolated using bacteriological method on  the second day of the study from clinical material obtained from sick people and environmental objects were detected in DIA at concentrations ≥ 3.1 × 104 microbial cells per milliliter (m.c./ml).Conclusion. The designed test system for dot-immunoassay using SNPs as a marker of specific antibodies for the detection of Y.  pseudotuberculosis in cultures isolated from swabs from vegetables and clinical material from patients, including those  with  mixed infection, allows us to  detect a specific corpuscular antigen with a high sensitivity (≥ 3.1 × 104 m.c./ml), providing express identification of isolated cultures at the initial stage of bacteriological study

IMPACT OF LEVOSIMENDAN ON RENAL FUNCTION IN COMPLEX TREATMENT OF ACUTE DECOMPENSATED HEART FAILURE

Background. Levosimendan infusion can be used in the treatment of patients with acute decompensated heart failure (ADHF) with a reduction in cardiac output and signs of severe congestion/pulmonary edema.Aim. To study impact of levosimendan on renal function in patients with ADHF with reduced systolic function.Material and methods. The study was a prospective, randomized trial. We enrolled 30 men (age 62.5 [55.8-69.3] years) hospitalized with ADHF with reduced systolic function (left ventricular ejection fraction <40%), increased level of brain natriuretic peptide (BNP>500 pg/mL) and systolic blood pressure >125 mmHg. All patients were randomized into 2 groups of 15 people each. In the first group, the patients received an intravenous infusion of levosimendan 0.1 μg/kg/min for 24 hour added to standard therapy. The second group received standard therapy.Results. 24-hour levosimendan infusion significantly increased the glomerular filtration rate levels from 65.4 [45.2-99.2] mL/min/1.73m2 at baseline to 79.0 [66.3-93.1] mL/min/1.73m2 at discharge (p= 0.011), greatly decreased serum creatinine from 1.17 [0.90-1.55] mg/dL at baseline to 1.01 [0.89-1.14] mg/dL at discharge (p = 0.009) and blood urea nitrogen and at the same time improved renal blood flow in patients with ADHF while there were no clinically significant changes in the studied parameters in the standard therapy group.Conclusion. Levosimendan had a positive effect on renal function in patients with ADHF with reduced systolic function

ASSESSMENT OF SITAGLIPTIN INFLUENCE ON ARTERIAL WALL STIFFNESS, RENAL FUNCTION AND RENAL CIRCULATION IN CARDIOVSCULAR PATIENTS WITH DECOMPENSATED TYPE 2 DIABETES

Aim. To study dynamics of stiffness parameters in various type arteries, renal function and renal circulation in arterial hypertension (AH) and ischemic heart disease (CHD ) with decompensated 2 type diabetes mellitus (DM2) on the sitagliptin therapy during 24 weeks.Material and methods. Totally 30 patients included, with decompensated DM2 (HbA1c >7%) and AH, most having also CHD. The dynamics of carbohydrate and lipid metabolisms were assessed, arterial wall stiffness in various structural and functional types, renal function and renal circulation at the background of sitagliptine therapy during 24 weeks.Results. There was no dynamics by the parameters of arterial wall stiffness in various types of arteries, as in renal function and renal circulation among the patients in sitagliptin (n=15) and comparison (n=15) groups. However in those achieved compensation of DM2 on sitagliptin (n=8) we found a decrease of stiffnes index β of brachial artery (muscular type) by 37% from baseline (p<0,01). There was no dynamics of stiffness in muscle-elastic or muscular types of arteries.Conclusion. In patients with hipertension disease and ischemic heart disease with DM2 on glucoselowering therapy there was no dynamics by the parameters of aortic, common caritid artery stiffness and renal circulation during 6 months followup. However on the therapy by dipeptidilpeptidase-4 inhibitor (sitagliptin) during 6 months and compensation of DM2 (HbA1c <7%) there was a decrease of brachial artery stiffness (muscular type)