A method of evaluating the toxicity of pertussis vaccine
(57) Abstract:The invention relates to medicine, more specifically to the field of Microbiology and vaccination and for the assessment of the toxicity of pertussis vaccine. The method is performed by immunization of experimental animals pertussis vaccine, pre-treated by low-frequency ultrasound (40-50 kHz), a day later mice decapitat, in the brain tissue to determine the number of histamine and when detecting it from a 22.4 µg/g and above is judged on the severity of the toxic effect of pertussis vaccines. The advantage of the invention is to simplify the method and shortening. table 2. The invention relates to the field of health, but rather to Microbiology and the vaccination and can be used to assess the toxicity of pertussis commercial vaccines manufactured in serial production.Analysis of data on complications in children, caused by a vaccine preparations, showed that in the first place by the number and severity of complications is adsorbed pertussis-diphtheria-tetanus vaccine (DPT), in which the main factor determining its negative characteristic is the pertussis component (Marshall L. csino. /Epidemiol. Rev., 1992, 14:243-287; Prins, G., Spellman, P. Pertussis. / Russ. the honey. Zhur., 1995, T. 1, No. 2, may). Severe complications induced by pertussis corpuscular vaccine, was the basis for the rejection of vaccination against pertussis in countries such as Japan, England, Sweden, and others (Stetler H. E. A. The benefits, risks from vaccination, and the judiciary. /J. Pediatr., 1985, T. 227, No. 692, S. 1289), but it has caused a mass outbreak of pertussis infection in unvaccinated children with a high percentage of severe forms, even fatal outcomes (Death from pertussis. - PPZS - centre for surveillance of infectious diseases.” Brit. Med. J. 1983, T. 286, No. 372, S. 1208, and others).Thus, the use of the pertussis vaccine can lead to serious post-vaccination complications a wide range of up to encephalopathy and encephalomyelitis, and the refusal of immunization - the possibility of the disease pertussis with the defeat of the vital functions of the body until death.To remedy the situation along with the development of vaccines with reduced side effects, the obvious relevance of search informative methods assessment criteria the safety of the pertussis vaccine, manufactured in serial production.One of the most serious proyavlyali you, encephalopathy, encephalomyocarditis, mility and others, which are based on damage to the functional structures of the nervous tissue (violation permeability hematoencephalic barrier, destruction of the myelin fibers of the cerebral cortex) (Cancerin A. H., Gervasia Century B. To the question of the pathogenesis of pertussis vaccine-related complications. In Proc. “Immunology of nervous and mental diseases”. - M., 1975, S. 276-280; Braginsky B. N. Vaccination neurological complications in children. /Nevropatol. and the psychiatrist., 1990; Prins, G., Spellman, P. Pertussis. /Russ. the honey. Zhur., 1995, T. 1, No. 2, may, and others).Conducted research on the medical-scientific and patent literature identified a number of ways to determine the damaging effect of pertussis vaccines on the nervous system of the body. So, A. A. Babchin in the article “Blanc-transformacija of peripheral blood lymphocytes with pertussis experimental allergic encephalitis (AAA)” (IN Coll. “Immunology of nervous and mental diseases”. - M., 1974, S. 276-280) describes a method of assessing toxicity of pertussis drugs on the nervous system by the method of besttransport of peripheral blood lymphocytes of Guinea pigs immunized with pertussis vaccines, where as mitogen IP the VA state method cytotoxic action of sensitized lymphocytes from animals grafted pertussis antigens in cell cultures of nervous tissue. M. M. Zotov proposes to use to assess the toxicity of pertussis drugs cytotoxic effect of serum and gamma-globulin fractions with pertussis, AAA identified through migration of macrophages (In Coll. “Immunology of nervous and mental diseases”. - M., 1974, S. 298-299). But the above methods do not give full information on the nature of the damaging effect of pertussis preparations are characterized by high variability, laborious research and the use of expensive reagents.The prototype of the present invention is the method described in the patent of the Russian Federation No. 2174229 (Moskalenko, E. P., Ilyin S. I., Uranowski C. F., “a Method for assessing the toxicity of pertussis vaccine”, BIPM No. 27, 2001). The method consists in the fact that the mice intraperitoneally injected with pertussis vaccine, pre-treated by low-frequency ultrasound (42-50 kHz) for 5 minutes. After 7 days mice decapitat in accordance with the “Rules of the experimental work on laboratory animals and collect brain tissue. Part of the brain is fixed in a 2% solution of camerahouse osmium (pH of 7.3) for dehydration. Then fabric sign in seonam microscope WANG-100 K. Detection of demyelination of myelinated fibers is one of the leading signs of encephalopathy reactions and indicates the severity of the toxic effect of pertussis vaccines on the nervous tissue of the host.This way peculiar to the following disadvantages: long term research, the complexity associated with the manufacture of sections of brain tissue, the use of expensive devices (electron microscope), is not available for practical health care.The aim of the present invention is to simplify the method of reduction of terms of research and expensive materials.This goal is achieved by immunization of experimental animals pertussis vaccine prepared according to the MRTU 42-263-68 (Technical conditions for the preparation of pertussis vaccine, approved by the Ministry of health of the USSR, 1968), pre-treated by low-frequency ultrasound (42-50 kHz) for 5 minutes at a dose of component 1.5 billion MT, a day later mice decapitat, in the brain tissue to determine the number of histamine and when detecting it from the 22.8 µg/g and above is judged on the severity of the toxic effect of pertussis vaccines on the nervous system of the body.DETAILED OP-263-68, containing 500 billion MT in 1 ml, treated with low-frequency ultrasound (42-50 kHz) with an amplitude of oscillations of the waveguide-disintegrator 10-30 microns at exposure exposure 5 minutes and left for 24 hours at a temperature of 3-6P. Then it is diluted with isotonic saline to a concentration corresponding to 1.5 billion MT in volume of 0.5 ml and subjected to immunization dose of 3-5 mice intraperitoneally. A day later mice decapitat in accordance with the “Rules of the experimental work on laboratory animals, collect brain tissue (cortex and white matter of cerebral hemispheres) and standard methods to determine the number of freely-active and lateral-associated histamine. The change in the amount of histamine in the brain tissue, as compared with the start of the pathological process, selected, guided by the data set out in the review article by Horunza T. A., Bardakjian E. A., Saakov B. A. “Mechanisms of experimental demyelination (In Coll. “Topical issues of immunology and immunopathology”, Rostov-on-don, 1975, S. 110-116), which showed that the accumulation of free-active and lateral-associated histamine has a damaging effect on the myelin causes significant activation of lysosomal apparire, what is the trigger mechanism in the induction of experimental allergic encephalomyelitis (EAE). Due to the fact that the degree of disturbance of vascular permeability increases, demyelination takes on a larger scale, which in turn exacerbates proteolysis, leading to the critical point of vascular permeability and, consequently, to massive cellular infiltration over time, coincident with the clinical manifestation encephalomyelitis.Treatment of pertussis vaccines to low-frequency ultrasound was used to provide the output of the substances responsible for encephalitogenic activity of bacterial cells, for the most complete assessment of the toxic properties of the tested drugs. Used low frequency ultrasound (42-50 kHz) with an amplitude of oscillations of the waveguide-desintegrator 10-30 μm, when exposure exposure to 5 minutes. The choice of this mode is due to the fact that low frequency ultrasound has significant advantages compared with high-frequency ultrasound, as it allows to reduce the acoustic power of impact, increasing the efficiency of research. In addition, low-frequency transformation of the energy of the acoustic heat allows p and lipoproteins, during ultrasonic impact (Braginsky F. I., “Quantitative regularities of ultrasound action on biomolecules and cells, and ultrasonic spectroscopy of biological structures”. Abstract. Prof. Diss., 1981, S. 39).For testing parameters of the proposed method were carried out relevant studies. In these experiments used the pertussis vaccine, prepared in accordance with MRTU 42-263-68 (Technical conditions for the preparation of pertussis vaccine, approved by the Ministry of health of the USSR, 1968) from industrial strains, obtained from the collection of the State Institute of standardization and control of medical and biological preparations. L. A. Tarasevich (,Moscow). Highly toxic pertussis vaccines were prepared from strains 688 and 703, the average toxicity of strains 345 and 222 and slightly toxic - from strain 187.The results of the study of the accumulation of histamine in the cortex and white matter of cerebral hemispheres experimental animals after 24 hours (1 day) after the introduction of subjects pertussis preparations showed that pertussis vaccines from strains 703 688, and induced the accumulation of histamine in the brain tissue of mice that were recorded from 31,8 to 34.4 µg/g, pertussis vaccine with the fry vaccine characterized by low toxicity, did not possess such properties (table.1). The amount of histamine in the brain tissue of animals immunizedthis vaccine differed slightly from the results obtained in the groups of animals inoculated with 0.15 M solution of sodium chloride (11.8 and 8.9 mg/g, respectively).It is obvious that the pertussis vaccine, causing the accumulation of histamine in the brain tissue, which is the starting mechanism in the induction of encephalitides disorders up to encephalomyelitis and encephalopathy, in the practice of commercial production of mono - and associated pertussis vaccine are to be excluded.On the basis of the conducted research revealed a number of distinctive features of the proposed method in comparison with the prototype, which is presented in table 2.Thus, as can be seen from table 2, the proposed method is compared with the prototype reduces to 7 times the period of analysis, in 42,6 times the cost analysis, but also greatly simplifies the complexity of the process. A method of evaluating the toxicity of pertussis vaccine by immunization of experimental animals, now ultrasound (42-50 kHz) for 5 min, after 24 h mice decapitat, in the brain tissue to determine the number of histamine and when detecting it from a 22.4 µg/g and above is judged on the severity of the toxic effect of pertussis vaccines on the nervous system of the body.
FIELD: medicine, psychiatry.
SUBSTANCE: one should isolate DNA out of lymphocytes of peripheral venous blood, then due to the method of polymerase chain reaction of DNA synthesis one should amplify the fragments of hSERT locus of serotonin carrier gene and at detecting genotype 12/10 one should predict the risk for the development of hallucino-delirious forms of psychoses of cerebro-atherosclerotic genesis.
EFFECT: more objective prediction of disease development.
FIELD: medicine, urology.
SUBSTANCE: one should conduct subcutaneous prevocational tuberculin test and, additionally, both before the test and 48 h later it is necessary to perform the mapping of prostatic vessels and at decreased values of hemodynamics one should diagnose tuberculosis. The information obtained should be documented due to printing dopplerograms.
EFFECT: more reliable and objective information.
1 ex, 1 tbl
FIELD: molecular biology.
SUBSTANCE: the suggested innovation deals with the fact that nucleic acids should be isolated directly out of the sample without pipetting stage but with the help of interconnected reservoirs being prepared beforehand. The above-mentioned vessels should be applied either separately or being interconnected according to standard microtitrating format. The sample should be mixed with a lyzing buffer and nucleic acids are bound with matrix in closed system including, at least, two interconnected reservoirs. Forced movement of sample's mixture and buffer back and forth from one reservoir into another one for several times through narrow passage provides their thorough intermixing. The method provides quick and safe isolation of nucleic acids.
EFFECT: higher efficiency.
44 cl, 4 dwg, 1 ex
FIELD: medicine, phthisiology, microbiology.
SUBSTANCE: diagnostic material is poured preliminary with chlorohexidine bigluconium solution, homogenized, kept at room temperature for 10-12 h and centrifuged. Precipitate is poured with Shkolnikova's liquid medium, incubated at 37oC for 3 days, supernatant part of Shkolnokova's medium is removed, fresh Shkolnikova's medium is added, and precipitate is stirred and inoculated on the dense cellular egg media. Sensitivity of the strain is determined in 3 weeks by the presence of growth in the control tube only. Invention provides enhancing precision and reducing time for assay. Invention can be used in assay for medicinal sensitivity of tuberculosis mycobacterium.
EFFECT: improved assay method.
FIELD: medicine, biotechnology, pharmacy.
SUBSTANCE: invention relates to agents used for treatment of pathological states associated with disorder of synthesis of neuromediating substances. Method involves the development of pharmaceutical composition and a method for it preparing. Pharmaceutical composition represents subcellular synaptosomal fractions: synaptic membranes, "light" synaptosomes and "heavy" synaptosomes prepared from gray matter of cerebral hemispheres from experimental animals based on the goal-seeking modification of humoral mediators of nerve endings transformed to synaptosomes in development and regression of malignant processes. The composition provides inhibiting the growth of tumor cells, to elevate span-life of patients with ascite Ehrlich's sarcoma, breast adenocarcinoma Ca-755, Wolker's carcinosarcoma-256.
EFFECT: valuable medicinal and anti-tumor properties of composition.
12 cl, 3 tbl, 3 ex
SUBSTANCE: method involves carrying out microscopic examination of blood serum samples taken from femoral vein and cubital vein. Femoral vein sample is taken on injured side. The examination is carried out before and after treatment. The blood serum samples are placed on fat-free glass slide in the amount of 0.01-0.02 ml as drops, dried at 18-30°C for 18-24 h. The set of pathological symptoms becoming larger or not changed after the treatment in comparison to sample taken before treatment, and morphological picture of samples under comparison taken from the cubital vein showing no changes or being changed to worse, the treatment is considered to be effective.
EFFECT: enabled medicamentous treatment evaluation in course of treatment to allow the treatment mode to be changed in due time; avoided surgical intervention (amputation); retained active life-style of aged patients.
FIELD: medicine, clinical toxicology.
SUBSTANCE: at patient's hospitalization one should gather the data of clinical and laboratory values: on the type of chemical substance, patient's age, data of clinical survey and laboratory values: body temperature, the presence or absence of dysphonia, oliguria being below 30 ml/h, hemoglobinuria, erythrocytic hemolysis, exotoxic shock, glucose level in blood, fibrinogen and creatinine concentration in blood serum, general bilirubin, prothrombin index (PTI), Ph-plasma, the state of blood clotting system. The state of every sign should be evaluated in points to be then summed up and at exceeding the sum of points being above "+20" one should predict unfavorable result. At the sum of "-13" prediction should be stated upon as favorable and at "-13" up to "+20" - prediction is considered to be doubtful.
EFFECT: higher accuracy of prediction.
2 ex, 3 tbl
FIELD: medicine, juvenile clinical nephrology.
SUBSTANCE: disease duration in case of obstructive pyelonephritis should be detected by two ways: either by detecting the value of NADPH-diaphorase activity, as the marker of nitroxide synthase activity in different renal department and comparing it to established norm, or by detecting clinico-laboratory values, such as: hemoglobin, leukocytes, eosinophils, urea, beta-lipoproteides, lymphocytes, neutrophils, the level of glomerular filtration, that of canalicular reabsorption, urinary specific weight, daily excretion of oxalates, arterial pressure, and estimating their deviation against average statistical values by taking into account a child's age.
EFFECT: higher efficiency of detection.
7 dwg, 1 ex, 6 tbl
FIELD: medicine, urology.
SUBSTANCE: the present innovation deals with differential diagnostics of prostatic cancer and other prostatic diseases at the stage of primary inspection. The method includes the detection of PCA and calculation of probability coefficient for prostatic cancer (PCC) by the following formula: where e - the foundation of natural logarithm (e=2.718…), PCA - the level of total blood PCA in ng/ml, V - patient's age in years. At PCC value being above 0.2 one should diagnose prostatic cancer and to establish final diagnosis one should perform polyfocal prostatic biopsy. The method enables to increase accuracy of diagnostics at decreased number of unjustified prostatic biopsies.
EFFECT: higher efficiency of diagnostics.
FIELD: medicine, biology.
SUBSTANCE: invention relates to nutrient medium used for accumulation of cells for the following cytological and/or immunocytochemical analysis carrying out. Invention relates to medium containing salts NaCl, KCl, anhydrous CaCl2, MgSO4 x 6 H2O, MgCl2 x 6 H2O, Na2HPO4 x 2 H2O, KHPO4, NaHCO3, and also glucose and Henx's solution, 10% albumin solution and polyglucin taken in the ratio 1:1:1. Invention provides enhancing the preservation of cells.
EFFECT: improved an valuable properties of nutrient medium.