Method for identifying forms of urogenital trichomonads
SUBSTANCE: method involves sampling a biological tissue, introducing a biologically active substance therein, keeping the prepared mixture in a thermostat at 37°C and incubating, performing microscopic examination for identifying the forms of trichomonads. The sampled biological tissue represents the urogenital mucosa that is incubated for 24 h in a nutrient medium. The prepared sediment is separated and added with the biologically active substance nitrosoguanidine in an amount of 500 mcg/ml. That is followed by incubating for 15 min at +37°C; a phosphate buffer with pH 5.6 heated to +37°C is added and centrifuged twice at 1000 rpm for 10 min. The material is placed into the nutrient medium for trichomonad growing, incubated for at least 24 hours, and microscopic examination is used to identify atypical nonflagellate and typical round and amoeboid forms of trichomonads.
EFFECT: using the declared invention enables the effective, fast and reliable identification of typical changed and atypical forms of trichomonads.
3 dwg, 3 ex
The invention relates to medicine, namely venereology, urology, obstetrics and gynecology, Parasitology, medical Microbiology.
The known method of identification of typical forms of Trichomonas, which after the receipt of pathological material from mucous membranes of the urogenital organs carry out sowing his method of inoculation in liquid nutrient medium, followed by incubation at +37°C for 3-17 days (Appendix №2 to the order of the Ministry of health of the USSR from July 12, 1985, No. 936, "guidelines on application of standardized clinical diagnostic and bacteriological studies in the diagnosis of gonorrhea and trichomoniasis", p.2.6).
The disadvantage of this method is that it is not possible to identify atypical forms of Trichomonas that, showing low metabolic activity, hardly propagated in nutrient media. Study long, not giving the possibility of rapid diagnosis.
A method for detecting Trichomonas forms, including blood cultures in a nutrient medium Terrace with the addition of inactivated for 5-10 minutes at a temperature of 56°C human serum or horse and antibiotics in the amount of 500 IU/ml Carried out the incubation of the culture for 2 days at 37°C. Then twice repeat these cycles with various�CNAME nutrient environments and exposure modes, then identify forms of trichomonads (Patent RF 2289813, IPC G01N 33/48, publ. 2006).
The disadvantages of this method are its complexity, length of performance, there is a risk for questionable or negative results due to the study of pathological material - blood, which is an unusual habitat for Trichomonas. Mode centrifugation 3000 rpm for 20-30 minutes is a gross stress factor leading to the destruction of the protozoa. With rounded and amoeboid forms of Trichomonas, and also other forms are probably not atypical cells, and variants simplest adaptation to changing environmental conditions.
The closest is a method for detecting Trichomonas forms (Patent RF 2262104, IPC G01N 33/48, publ. 2005), including the capture of biological material, the addition of biologically active substances, exposure of the mixture in a thermostat at 37°C, followed by incubation for 24-48 h, the conducting microscopic examination for the detection of Trichomonas forms. As a result of repeated treatment every 12 hours a new dose of a biologically active substance (enzyme) mixture is the destruction of red blood cells, microscopically manifested in the form of fragments of cellular elements, among which a notable modified and atypical forms of Trichomonas.
The object of the invention is to eliminate these drawbacks, increasing the reliability of detection in pathological material Trichomonas forms, reducing the time needed to obtain result, detection of the modified typical and atypical forms of Trichomonas by adding nitrosoguanidine to the precipitate overnight culture of protozoa with subsequent laundering and the addition of nutrient medium, which causes changes in the nucleus (polymorphism: rounded, elongated - 2/3 of the length of a simple, curved, reduced in size of the nucleus); a supporting unit (axostyle) - shortening, thickening, curved axostyle, no spinules; the loss of the organs of movement - flagella; changing the metabolism of the simplest - damage hydrogenosome (analogues of mitochondria), activation Pino - and phagocytosis, the formation of giant vacuoles.
To do this, in the method for detecting Trichomonas forms, including z�their biological tissue, introduction of biologically active substances, exposure of the mixture in a thermostat at 37°C, followed by incubation, conducting microscopic examination for the detection of atypical forms of Trichomonas proposed as a biological material to use the scrapings from the mucous membranes of the urogenital organs. This biological tissue was incubated in a day in a nutrient medium, separating the resulting precipitate and add to it as biologically active substances nitrosoguanidine in the amount of 500 μg/ml, incubated for 15 min at +37°C, add preheated to a temperature of +37°C phosphate buffer with a pH of 5.6 followed by a double by centrifugation at 1000 rpm for 10 min, then placed in a nutrient medium for the cultivation of Trichomonas incubated for at least a day and microscopy reveal atypical besthomemovies and typical rounded and amoeboid forms of trichomonads.
Fig.1 shows basiguttata form Trichomonas (example 1); Fig.2 - rounded Trichomonas (example 2); Fig.3 - amoeboid form of Trichomonas (example 3).
The proposed method provides culture diagnosis typical modified and atypical forms of Trichomonas. This occurs under the action of nitrosoguanidine, which is a chemical mutagen (stress factor) that causes the activation of Pinot and Fugazi�oz, active fixation simplest to the surface of epithelial cells and to each other, the formation of colonies of protozoa for the purpose of self-preservation.
The method is as follows.
Material taken from the vaults of the vagina (in women) or urethra (men) sterile cotton micronova swab was immersed in 5 ml of nutrient medium for the cultivation of Trichomonas, the swab was rinsed in the environment. Were incubated for 24 h at +37°C. When grown on liquid nutrient media urogenital Trichomonas give demersal growth in the form of dense whitish precipitate.
For the preparation of nutrient media used "the Basis of the nutrient medium for the isolation of vaginal Trichomonas (dry) (production FSUE "NPO "Microgen" MOH", cat. No. C01). The composition of the basis in g/l: extract of fodder yeast, D-maltose, sodium chloride, potassium chloride, sodium carbonate (pH 6,3±0,3). Method of preparation: 28.5 g of the preparation is stirred in 1 l of distilled water, boiled for 1-2 minutes and poured into vials. The medium was sterilized by autoclaving at t +112°C for 20 min To a cooled to t +45-50°C culture medium was added 20% bovine serum, thoroughly stirred and poured into 5 ml sterile tubes. The surface of the medium in test tubes filled with sterile paraffin oil layer height of 3-5 mm. Tubes with nutrients such a�filled, the medium was incubated at t +36±1.0°C for 22-24 h to control the sterility of the environment.
To obtain atypical forms of urogenital Trichomonas precipitate overnight culture was adjusted with physiological saline to a volume of 1.0 ml and was added nitrosoguanidine in the amount of 500 μg/ml. According to the literature on the effects of the mutagen on the simplest indicates of their 50% death in culture (Pimenov M. N., Grechushkina N. N., Azov L. G., Netrusov A. I. a Guide to practical exercises in Microbiology/ edited by Egorova N. With. - 3rd ed.; revised and enlarged extra - M.: Publishing house of IPA, 1995. - Pp. 173-179).
After the end of exposure time (15 min) at a temperature of +37°C was added 3 ml preheated to a temperature of +37°C phosphate buffer with a pH of 5.6 followed by a double by centrifugation at 1000 rpm for 10 min.
After washing the culture of protozoa was added 5.0 ml of the nutrient medium for the cultivation of Trichomonas and were incubated for 24 h at +37°C. the cultivation mutated culture Trichomonas vaginalis allows to identify various typical modified and atypical (besthomemovies) shape in the form of near-bottom dense whitish precipitate.
The presence of the above forms of Trichomonas allows to confirm the diagnosis of trichomoniasis, and in their absence to exclude this diagnosis.
Patient M., aged 45, was admitted with a diagnosis of trichomoniasis.
Investigated the pathological material from the posterior and lateral vaginal fornix. Conducted researched�e of the proposed method.
After being exposed nitrosoguanidine to precipitate overnight culture of Trichomonas detected besthomemovies forms of trichomonads.
The diagnosis was confirmed - urogenital trichomoniasis.
Treatment: metronidazole well-known scheme. After treatment is complete recovery.
Patient O., aged 27, was admitted with a diagnosis of trichomoniasis.
Investigated the pathological material from the posterior and lateral vaginal fornix. After being exposed nitrosoguanidine to precipitate overnight culture of Trichomonas detected rounded Trichomonas.
The diagnosis of urogenital trichomoniasis.
The patient was treated with metronidazole to cure.
Patient F., 30 years.
Investigated the pathological material from the posterior and lateral vaginal fornix.
By the above procedure is carried out the impact nitrosoguanidine to precipitate overnight culture of trichomonads found amoeboid forms of trichomonads.
The diagnosis of urogenital trichomoniasis.
The patient was treated with metronidazole.
The proposed method was applied in 2245 patients, the identification of the forms allowed to pick up adequate drug therapy and to achieve a complete etiological cure patients.
This method is of great importance for the diagnosis of urogenital trichomoniasis when infection occurs asymp�UMNO and latent, while the pathogen has amastigote (basiguttata) form, which is difficult to detect with conventional culture methods of diagnosis. With the help of this technique confirmed the existence of atypical (besthomemovi) forms of protozoa identified the phenomenon of dropping flagella and transformation of Trichomonas in besthomemovies form (amastigote).
A method of detecting Trichomonas forms, including the taking of biological tissue, the introduction of biologically active substances, exposure of the mixture in a thermostat at 37°C, followed by incubation, conducting microscopic examination for the detection of Trichomonas forms, characterized in that as a biological tissue carry out the extraction of the material from the mucous membranes of the urogenital organs, which were incubated in a day in a nutrient medium, separating the resulting precipitate and add to it as biologically active substances nitrosoguanidine in the amount of 500 μg/ml, incubated for 15 min at +37°C, add preheated to a temperature of +37°C phosphate buffer with a pH of 5.6 followed by a double by centrifugation at 1000 rpm for 10 min, then placed in a nutrient medium for the cultivation of Trichomonas incubated for at least a day and microscopy reveal atypical besthomemovies and typical rounded and amoeboid forms of trichomonads.
SUBSTANCE: method involves measuring pregnant women's blood anti-cytomegalovirus antibody titre and glutathione reductase activity in erythrocytes of the obstetric patient suffering the cytomegaloviral infection. If the anti-cytomegalovirus antibody titre is 1:1600, whereas the measured glutathione reductase activity is below 4.48±0.22 units/gHb, the aggravated cytomegaloviral infection is diagnosed.
EFFECT: higher diagnostic technique.
SUBSTANCE: group of inventions relates to medicine, cosmetology, production of food products, vitamins, food supplements, drugs and describes versions of device for realisation of non-invasive potentiometric determination of oxidant/antioxidant activity of biological tissues, which includes device for measuring potentials and double-sided electrode, made in form of plate with similar working surface, covered with electricity-conducting gel, containing mediator system. Electrodes are fixed on biological tissue in such a way that one working surface, playing role of measuring electrode, is in direct contact with biological tissue via gel, second working surface pale role of comparison electrode. Electrodes contact with each other via gel, with oxidant/antioxidant activity being determined by formulae with application of difference between final and initial potentials.
EFFECT: simplification, as well as increase of accuracy and reliability of determination, is achieved.
14 cl, 3 tbl, 4 dwg
SUBSTANCE: invention represents a method for the prediction of preeclampsia in the second trimester of pregnancy by blood examination, differing by the fact that the activity of acid and neutral proteinases is measured in blood serum of the women 7-8 weeks pregnant; if the activity of acid proteinases is more than 5.6 mcmole/l, while the activity of neutral proteinases is more than 3.9 mcmole/l, the preeclampsia progression in the second trimester of pregnancy is predicted.
EFFECT: higher accuracy and specificity of the method for the prediction of gestational toxicosis.
SUBSTANCE: invention relates to medicine, namely to a method of automated morphometric myelofibrosis diagnostics. The method essence consists in the fact that overview images of zones with different optical properties with determinable fibrous and heamopoietic properties of a biological tissue are performed. Ratios of areas of the said zones of at least three paraffin cuts of trepanobiopsy samples are calculated. The coefficient (Cop) is calculated as the ratio of the fibrous tissue area to the area of the heamopoietic tissue by formula. If the value Cop ≥14.5%, myelofibrosis is diagnosed.
EFFECT: application of the claimed method makes it possible to increase the accuracy and improve the efficiency of myelofibrosis diagnostics.
7 cl, 1 tbl, 4 dwg, 1 ex
SUBSTANCE: testicular germ cells are measured quantitatively. That is ensured by 50-day oral administration of selexen and ascorbic acid into male white rats in doses 1.5 and 500 mg/kg of animal's body weight respectively once a day. 14 days later, administering the selenium-containing biocomplex is accompanied by the 30-minute daily exposure to microwave radiation at 42 GHz (λ=7.1 mm) for 30 days. Once the experimental exposures are completed, the corrective properties of the biocomplex as having an effect on the morphofunctional state of epididymal sperm cells are assessing by formula: MFSI=A+B, wherein MFSI is a morphofunctional state index, A is a portion of normal sperm cells in relation to the reference, and B is a portion of moving sperm cells in relation to the reference. If the MFSI value is 1.3 or more, the spermatogenesis correction is considered to be ineffective, while the MFSI value being more than 1.3 shows the effective spermatogenesis correction if exposed to microwave radiation.
EFFECT: invention enables assessing the spermatogenesis correction efficacy with underlying administration of the biocorrector.
2 tbl, 3 dwg, 3 ex
SUBSTANCE: invention relates to field of medicine, in particular hepatology. Method of quantitative estimation of histological activity in liver biopsy materials in case of chronic diffuse liver diseases results in protocol of counting in absolute numbers, percent and points of pathological signs, identified in the course of morphological analysis of liver biopsy materials from patients with chronic diffuse liver diseases, by sections: necrosis of hepatocytes, globular and atomised fatty hydropic degeneration of hepatocytes, small cell dysplasia of hepatocytes, portal, periportal, intralobular and central infiltration. After that conversion of absolute values into point system is performed by means of said protocol, and histological activity is determined by number of points.
EFFECT: method makes it possible to create quantitative unification of obtained results in case of said diseases.
FIELD: veterinary science.
SUBSTANCE: method includes detection of leukocytes, protein, haemoglobin in faeces, and PH-reaction of faeces. A sample of faeces is dissolved in distilled water and applied drop by drop onto appropriate test fields of a test strip designed for urine testing. By variation of colour within 1 minute the reaction is assumed as positive. If pH is less than 7.0 or more than 7.5, and faeces contain soluble protein, haemoglobin and leukocytes at the same time, availability of the inflammatory process in the intestines is confirmed.
EFFECT: invention makes it possible to quickly and accurately diagnose inflammatory process in intestines.
2 cl, 2 ex
SUBSTANCE: invention relates to medicine and is intended for the treatment of patients with the perforation of gastric and duodenal ulcer. A laparotomy is performed, operative aid concerning the ulcer perforation is realised, during the operation an exudate is sampled from the abdominal cavity, pH of the exudate is determined. If Ph of the said material is 6.6 and lower, a conclusion about the presence in it of anaerobic flora is made and serves the basis for the administration of an antibacterial preparation, influencing the anaerobic flora, into the scheme of initial antibacterial therapy.
EFFECT: method makes it possible to administer specific antibacterial therapy without waiting for culture results.
SUBSTANCE: method involves the integrated clinical-laboratory and microbiological studies of patient's urogenital discharge to determine an agent's cytomorphotype as shown by culture analysis and evaluating a degree of manifestation of the patient's urogenital inflammation or leucocytosis.
EFFECT: invention enables improving the quality of laboratory diagnostics, provides establishing the accurate diagnosis that improves the clinical effectiveness, and reduces the rate of complication accompanying chronic trychomonad infection.
1 tbl, 2 ex
SUBSTANCE: invention refers to medicine, namely to a method for producing cells and non-cell elements of an atherosclerotic plaque. The substance of the method consists in the fact that a balloon angioplasty is followed by washing the cells and non-cell elements; the produced suspension of the cells and non-cell elements is centrifuged; a supernatant fluid is removed; the cells and non-cell elements are re-suspended; the suspensions of the cells and non-cell elements are placed into charged cytobuckets, which are centrifuged; the produced preparations are used for further analysis.
EFFECT: using the declared method provides producing the patient's cells and non-cell elements of the atherosclerotic plaque after the balloon angioplasty for the main indication.
5 cl, 10 dwg, 2 ex
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.