Method for determining individual sensitivity of human genome to radon exposure
SUBSTANCE: determining individual genome sensitivity to radon exposure is ensured by genetic blood test to identify predisposing and protective genotypes: marker Arg280His of gene XRCC1 - predisposing genotype Arg/Arg, protective genotype Arg/His; marker Argl94Trp of gene XRCC1 - predisposing genotype Arg/Arg, protective genotype Arg/Trp; marker Asnl48Glu of gene APE1 - predisposing genotype Glu/Glu, protective genotypes Asn/Asn, Asn/Glu; marker A2455G of gene CYP1A1 - predisposing genotype A/G, protective genotypes A/A and G/G; a deletion marker in gene GSTM1 - predisposing genotype o/o, protective +. High individual sensitivity to high radon dose is stated by observing the quantitative prevalence of predisposing genotypes or the equal quantity of predisposing and protective genotypes. High individual resistance to high radon dose is determined by the quantitative prevalence of protective genotypes.
EFFECT: use of the method allows estimating genetically determined predisposition to formation of high level of chromosomal aberrations even before exposing to a radiating factor.
5 tbl, 2 ex
The invention relates to the field of medicine, human genetics and can be used to assess individual radiosensitivity of the human genome (where live or work in conditions of exposure to high doses of radon).
There is a method of determining the genotoxic sensitivity to the complex chemical mutagens, manifested in the form of a certain level of chromosomal aberrations (CA), applicable for the assessment of professional suitability of individuals to work in environments with increased hazard for genetic markers (A.S. SU 1621718 A1, CL G01N 33/48, publ. 22.04.1988). The essence of the method lies in the fact that the Respondent take the blood from the cubital vein, cultivate her 48 hours in the presence of phytohemagglutinin, accumulation of chromosomes in metaphase use treatment by colchicine. This is followed by hypotonic treatment and fixation of the cells. The resulting suspension dig on a chilled glass slides and dried in air. With the help of the method of differential staining of chromosomes reveal extreme variants of polymorphism. For this purpose, preparations stained by the dye from the Institute after the preliminary denaturation in a saturated solution of Ba(OH)2and incubation in standard saline solution of 2×SSC at 60°C. the Evaluation segments performed semiquantitative method on a 5-point system. For ekström the global options accept options: 1 or 5 points, percentagesa inversion. The presence of the genotype of these options is associated with a high frequency of chromosomal abnormalities. Thus, in the presence of the genotype extreme variations of heterochromatin judged unsuitable for persons working in enterprises with high toxic hazard.
There is also known a method of determining individual sensitivity and suitability of individuals to work in the conditions of increased coal dust (patent RU 2316764 C1, CL G01N 33/48, publ. 10.02.2008). The essence of the method lies in the fact that the subjects for the study take 0.5-1.0 ml of blood from the cubital vein. Spend the cultivation of lymphocytes and receive chromosome by a standard method. Drugs sostarivayut" in thermostat at 37°C for 24 hours. The staining perform drawing on the preparation of silver nitrate and define the following cytogenetic indicators: total activity Adair (Ag-positive agricoevrazia regions of chromosomes), the number Adair, argentopyrite D-chromosomes, argentopyrite G-chromosomes, associative index, the number of associations per cell with associations, the number of associate of acrocentrics in a cage with associations. Based on the totality of the resulting figures calculate the confidence level high level HA and when it is 50% or less define the procedure the regional suitability of the test to work in the conditions of increased coal dust.
It should be noted that these methods imply different approaches to predict the level of CA when exposed to chemical compounds. Known methods of biological indication of radiation effects, allows to confirm the fact of oblojennosti and its degree of severity, based on the determination in peripheral blood lymphocytes of the subject HA dicentrics type and E-37-rozetka-forming lymphocytes (patent RU 2126156 C1, CL G01N 33/53, publ. 10.02.1999).
The closest way to determine individual radiosensitivity can serve as a method of rapid detection of irradiated patients with elevated frequencies of HA (patent RU 2141 658 C1, CL. G01N 33/48, publ. 20.11.1999). As a marker to identify patients with elevated frequencies of HA after radiation treatment using anomalies of interphase nuclei of lymphocytes type "tails" in the blood smears. With the increased frequency of occurrence of such cells (0.8% and above) identify patients with a high incidence of HA. The disadvantages of this method include the lack of prognostic significance, because in this way it is not about the risk of accumulation of chromosomal abnormalities, not about the definition of individual genome sensitivity to radiation exposure, but only about the display - identifying individuals with elevated levels of chromosomal mutations after radiation the frame exposure. This significantly narrows the scope and informative way.
Due to the successes of molecular medicine and development of an adequate molecular-genetic methods in recent years has made possible the identification of the relationship of genetic polymorphisms of DNA repair enzymes and biotransformation of xenobiotics with the response to the influence of environmental factors. Currently found potential genes-candidates, which may affect the frequency of spontaneous and induced chromosomal damage in humans. It is established that the null genotypes for glutathione-S-transferase (GSTM1 and GSTT1) is associated with higher levels of HA induced by mitomycin C (Grigoriev S.A. Study of genetically determined sensitivity to the effects of environmental mutagens in induced mutagenesis in humans. Abstract. Diss. M, 2007). Shown an increase in the frequency of the ring and dicentrics chromosomes in carriers of XRCC1 280 His/His in conditions of exposure to indoor radon (Kiuru, A., Lindholm C., Heilimo I. et al., 2005. Inflluence of DNA repair gene polymorphisms on the yield of chromosomal aberrations // Environmental and molecular mutagenesis. V.46 1.3. P.198-205). In vitro experiments have shown that individuals with the wild type XRCC1 194 Arg/Arg demonstrate a greater number of chromosome breaks per cell in the processing of cell cultures by bleomycin and tiolovymi the epoxides of benzo[a]pyrene (Wang y, Spitz M.R., Zhu Y. et al., 203. From genotype to phenotype: correlating XRCC1 polymorphisms with mutagen sensitivity // DNA Repair (Amst). 2 (8). P.901-908). However, these studies only recognise the contribution of individual genotypes in the formation of chromosomal aberrations, not offered specific ways of using the results to determine the individual sensitivity of the person to the action of mutagens.
The main objectives of the proposed invention are: to provide a method for determining individual sensitivity of the human genome to the effects of natural radiation (radon) and increase the prognostic value by engaging in the forecast system molecular genetic markers (genes, DNA repair enzymes and biotransformation of xenobiotics).
This is due to the fact that in the proposed method of determining individual sensitivity of the human genome to the effects of radon, including the exploration of human blood and the determination of individual level HA, identify predisposing and protective genotypes of candidate genes: marker Arg280His gene XRCC1 - predisposing genotype Arg/Arg, protective genotype Arg/His; marker Arg194Trp gene XRCC1 - predisposing genotype Arg/Arg, protective genotype Arg/Trp; marker Asn148Glu gene ARE - predisposing genotype Glu/Glu, protective genotypes Asn/Asn, Asn/Glu; marker A2455G gene CYP1A1 - predisposing genotype A/G, protective the genotypes a/a and G/G; marker deletion in the gene GSTM1 - predisposing genotype o/o protective +, and make a conclusion about the high individual sensitivity to the action of high doses of radon in the quantitative prevalence of predisposing genotypes or equal to the number of predisposing and protective genotypes, and the high individual resistance to the effects of radon in the quantitative predominance of protective genotypes.
The novelty of this approach lies in the development of complex molecular-genetic characteristics, assessment of the totality which is necessary and sufficient to determine the individual sensitivity of the human genome to the effects of radon (in doses above 200 Bq/m3).
First established the role of the polymorphism of some genes biotransformation of xenobiotics (GSTM1 and CYP1A1) in the formation of chromosome aberrations in conditions of chronic exposure to high doses of radon. Among a wide range of known polymorphic genes reparations selected markers associated with an increased risk of chromosomal abnormalities in terms of the natural radiation - Arg280His Arg194Trp and XRCC1 gene, Asn148Glu gene ARE.
Assessment of the totality of gene polymorphisms of DNA repair enzymes and biotransformation of xenobiotics used in the proposed method, has a high efficiency and accuracy determined by the ü potential risk of formation of chromosome aberrations in persons, exposed to natural radiation. This can be used to assess individual sensitivity to the action of radon in residential buildings, and industrial conditions, such as miners, working in conditions of exposure to very high doses of radon.
The method is performed in the following sequence:
1. Form groups of observations.
For this form two groups of donors with high (above average) and low levels of HA formed under conditions of chronic high exposure to radon. In our study, were surveyed Teens (211 people)living in the boarding school. It was experimentally found that the concentration of radon in indoor air boarding school exceeds the permissible level for operating buildings (up to 200 Bq/m3and reaches 583 Bq/m3in the winter, and 334 Bq/m3during the spring period.
The choice of teenagers is justified by the fact that in this case, minimizing the influence of factors such as bad habits, chronic diseases and the impact of professional contact with the production of harm. In addition, compact when all of the sample (living together in a boarding school for at least 3 months prior to study) provides maximum commonality in power settings and conditions of accommodation is found. All donors had no chronic diseases, were non-smokers. In the survey did not include children receiving medical treatment, and also held x-ray examination within 3 months. to collect the material. For each subject was decorated Protocol informed consent signed by the parents or persons performing the guardianship of minors. Age and sex characteristics of the groups are presented in table 1.
Preparations preparations of metaphase chromosomes was performed using standard polymicrobial cultivation of lymphocytes [Hungerford R.A., 1965. Leukocytes cultured from small inocula of whole blood and the preparation of metaphase chromosomes by treatment with hypotonic KC1 // Stain Techn. Vol.40. P.333-338]. Cultured whole venous blood, which was taken from the cubital vein. In the culture vial was placed 0.5 ml of blood, 0.1 ml of phytohemagglutinin (Paneco), 6 ml of medium RPMI 1640 (Paneco), 1.5 ml of fetal calf serum. Duration of cultivation was 48 h For 2 h before fixation in culture was added colchicine at a final concentration of 0.5 μg/ml At the end of cultivation, the cells were treated with hypotonic solution of 0.55% KCl for 10-15 min at 37°C. Fixation was performed in 3 shifts chilled ethanol-acetic latch (3:1). The cell suspension was excavated on a clean, chilled, moistened with water slides. PR is parathas encrypted and were stained with 2%dye solution from the Institute (Merk). Accounting HA conducted without karyotyping. The selection of metaphases to be included in the analysis, and criteria for the registration of cytogenetic damage was consistent with generally accepted recommendations [Bochkov N.P., Kuleshov, NP, Zhurkov B.C., 1972. The analysis of spontaneous chromosomal aberrations in cultures of human leukocytes // Cytology. T. No. 10. Pp.1267-1273]. Viewed 100-300 metaphases. Evaluated the frequency of cells with HA.
The average frequency of HA in the examined donors entire sample was and 5.30±0,16%. It significantly significantly exceeds the value of the base background levels for the region (2,86%), which confirms the fact genotoxic effects of radon. It is notable that, despite the similarity factors: the intensity of radon exposure, age, nutrition, health, 43% of children demonstrate particularly high levels of chromosomal aberrations. The 91 persons frequency of cells with HA was above average and was 7,42±0,18%. Obviously, the reasons for this increased sensitivity is genetically determined. Of these donors, and was formed by experienced group. In the comparison group included 120 people. The average frequency of cells with HA in this group was below average and was 3,48±0,12%. The difference between these groups was statistically significant (p<0,01).
2. Form a table of the relative risk.
Conduct frequent comparison the t allele genotypes and combinations of genotypes of polymorphic markers of candidate genes in the experimental and control groups. The comparison of the frequency is performed according to the criterion χ2with the amendment of Yates with SPT Statistica 6.0. The relative risk and confidence interval are calculated according to the formula:
wherea+0.5 to the number of donors with high levels of HA of native speakers of a given gene/genotype; b+0.5 to the number of speakers of a given gene/genotype with low levels of HA; c+0.5 to the number of donors with high levels of HA without this gene/genotype; d+0.5 to the number of donors with low levels of HA without this gene/genotype (adjusted for the small number of observations).
To construct a confidence interval, we take the value L=lnOP, which approximately can be considered normally distributed with a mean of:
and standard deviation:
The lower and upper confidence bounds of the index, between which the rates are with probability 1-a, respectively values:
where- value of student's criterion, the corresponding error probability α; in particular, when constructing a 95% confidence interval, α=0.05 and; when 99% on the credentials of the interval α=0.01 and ; and at the 99,9% confidence interval, α=0.001 and.
As the lower and upper confidence limits take antilogarithm values fromand:
If the confidence interval does not include the value of 1.0, then the feedback effect of the disease is considered statistically significant.
The value of the relative risk for all cause genotypes are shown in table 2. With a relative risk of more than 1.5 conclude about predisposition, with a relative risk of less than 0.7 is about genetic resistance to the formation of high levels of chromosomal disorders.
3. Form the table protective and predisposing genotypes (table 3).
4. Detailed description of the method.
The examined So 15 years in a sterile tube type "vacutainer" (with EDTA) was taken by venous blood in a volume of 1.5 ml DNA was isolated using phenol-chloroform extraction. For genotyping of nucleotide substitutions A2455G gene CYP1A1, Arg280His Arg194Trp and XRCC1 gene, Asn148Glu gene ARE used PCR and a set of reagents developed NPF Liteh" (Moscow). Detection of mutations in the genes CYP1A1 (TS), GSTM1 (deletion) was performed in accordance with the instructions of the reagent kits manufacturer (LLC Siring", Novosibirsk, Russia). Amplification wire is whether using amplifier "Terzic". Electrophoretic separation of the amplified fragments was performed in 3% agarose and subsequent color bromide by ethidium. Obtained values of the genotypes and alleles are presented in table 4. Established 6 predisposing genotypes. Based on the obtained results the conclusion about the existence of high-risk genetically determined chromosomal abnormalities when exposed to radon. This conclusion was confirmed by the fact that when pathogenetic analysis of the subject So the level of CA aberrations was 6.5%.
The examined So 11 years typing of polymorphic markers was performed as described in example 1. The results are presented in table 5. Identified the prevalence of protective genotypes (4 protective and 2 predisposing genotype). The conclusion of genetically determined resistance to the formation of chromosomal aberrations in terms of radon exposure. This conclusion was confirmed by the fact that the cytogenetic analysis of surveyed So the real level of HA was 3.5%.
5. The effectiveness of the method.
Determined polymorphic genes-candidates in the comparison group in adolescents exposed to chronic exposure to radon. Found that out of 120 people with a relatively small frequency HA (3,48±0,12%)) in 22 adolescents had dominated retrypolicy genotypes or equal to the number of predisposing and protective genotypes. This suggests that their individual sensitivity to radon exposure is increased. The average frequency of HA from them (3,84%±0,35%) was higher than the other 98 of 120 adolescents with prevalence of protective genotypes and with individual resistance to radon (3,38±0,78%). It is obvious that with increasing time of exposure to radon frequency HA 98 adolescents with genetically determined individual resistance will increase to a lesser extent than in 22 adolescents with genetically determined individual sensitivity. Thus, the group with a heightened sensitivity to the action of radon (91 people), formed using a known method of analysis HA, increased by 22 people through the use of the proposed method, which includes the determination of polymorphic candidate genes. That is, the efficiency of the method was increased by 24.2%.
The proposed method has greater prognostic significance, because it allows the assessment of a genetically determined predisposition to the formation of elevated levels of chromosomal aberrations before exposure to the radiation factor.
|Age and sex characteristics is istica groups|
|Index||Experimental group (donors with a high level of chromosomal aberrations)||The comparison group (donors with a low level of chromosomal aberrations)|
|The risk of high levels of chromosomal aberrations associated with the genotypes of the major candidate genes|
|Polymorphic marker||Genotype||The relative risk (RR and 95% CI)|
|Arg/Arg||of 11.26 [6,92-18,31]|
|Arg280His gene XRCC1||Arg/His||0, 09 [0,05-0,15]|
|Arg194Trp gene XRCC1||Arg/Trp||0,27 [0,16-0,45]|
|Asn148Glu gene ARE||Asn/Glu||of 0.37 [0.14 to 0,94]|
|A2455G gene CYP1A1||A/G||3,11 [1,16-8,30]|
|TS gene CYP1A1||T/C||0,57 [0,22-1,49]|
|C/C||to 2.29 [1,51-3,49]|
|a deletion in the gene GSTM1||0/0 (deletion)||3,00 [1,56-5,73]|
|0/+ and +/+||0,33 [0,17-0,64]|
|Protective and predisposing genotypes high-risk chromosomal abnormalities from donors exposed to indoor radon|
|Polymorphic markers||Predisposing genotypes||Protective genotypes|
|Arg280His gene XRCC1||Arg/Arg||Arg/His|
|Arg194Trp gene XRCC1||Arg/Arg||Arg/Trp|
|Asn148Glu gene APE1||Glu/Glu||Asn/Asn, Asn/Glu|
|A2455G gene CYP1A1||A/G||A/A, G/G|
|TS gene CYP1A1||T/T, C/C||T/C|
|a deletion in the gene GSTM1||0/0 (deletion)||+|
|Results genotyping 1|
|Polymorphic marker Arg280His gene XRCC1||Arg/Arg||of 11.26|
|Polymorphic marker Arg194Trp gene XRCC1||Arg/Arg||3,75|
|Polymorphic marker Asn148Glu gene ARE||Glu/Glu||2,30|
|Polymorphic marker A2455G gene CYP1A1||A/G||3,11|
|Polymorphic marker TS gene CYP1A1||T/T||1,76|
|Polymorphic marker deletion in the gene GSTM1||0/0||3,00|
|Results genotyping 2|
|Polymorphic marker Arg280His gene XRCC1||Arg/Arg||of 11.26|
|Polymorphic marker Arg194Trp gene XRCC1||Arg/Arg||3,75|
|Polymorphic marker Asn148Glu gene ARE||Asn /Glu||0,37|
|Polymorphic marker A2455G gene CYP1A1||A/A||0,32|
|Polymorphic marker TS gene CYP1A1||T/C||0,57|
|Polymorphic marker deletion in the gene GSTM1||+||0,33|
The method of determining individual sensitivity of the human genome to the effects of radon, including the identification of polymorphic markers of candidate genes by genetic studies of human blood, characterized in that determine predisposing and FR is active genotypes of candidate genes: marker Arg280His gene XRCC1 - predisposing genotype Arg/Arg, protective genotype Arg/His; marker Arg194Trp gene XRCC1 - predisposing genotype Arg/Arg, protective genotype Arg/Trp; marker Asnl48Glu gene ARE-predisposing genotype Glu/Glu, protective genotypes Asn/Asn, Asn/Glu; marker A2455G gene CYP1A1 - predisposing genotype A/G, the protective genotypes a/a and G/G; marker deletion in the gene GSTM1 - predisposing genotype o/o protective +, and make a conclusion about the high individual sensitivity to the action of high doses of radon in the quantitative the prevalence of predisposing genotypes or equal to the number of predisposing and protective genotypes, and the high individual resistance to the effects of high doses of radon - in the quantitative predominance of protective genotypes.
SUBSTANCE: patient examination involves visual evaluation of a periodontal pocket depth, a tooth mobility degree, gum tissue state with using periodontal indexes. X-ray examination aims at evaluating an alveolar bone resorption degree. It is followed with biochemical blood analysis for the parathormone and calcitonin concentrations. If simultaneously observing the blood calcitonin concentration less than a lower physiological norm, while the parathormone concentration exceeding an upper physiological norm, the presence of chronic aggressive generalised periodontitis is concluded.
EFFECT: use of the technique allows advanced detection of chronic aggressive generalised periodontitis, early identification of said disease among the other inflammatory diseases of periodontium at initial morbidities.
3 dwg, 2 tbl, 2 ex
SUBSTANCE: biological tissue is crushed, processed twice for 30 minutes with portions of ethyl acetate, weight of each twice exceeding weight of a biological object; prepared extractions are combined, filtered through anhydrous sodium sulphate; a solvent is evaporated from the filtrate; the residue is dissolved in acetonitrile; the prepared solution is watered down in the volume ratio 1:4, extracted twice in portions of chloroform, volume of each being equal to volume of a hydrophilic layer; the chloroform extracts are combined, steamed to a dry residue; the residue is dissolved in mixed solvents hexane-dioxane-propanol-2, cleaned in a silica gel column L 40/100µ with using a mobile phase hexane-dioxane-propanol-2; eluate fractions containing an analysed substance are combined; the eluent is evaporated; the residue is dissolved in mixed solvents hexane-dioxane-propanol-2 and analysed by a HELC method in a column of dimensions 64×2 mm filled with the sorbent Silasorb 600 with using a mobile phase hexane-dioxane-propanol-2 and a UV detector.
EFFECT: invention allows higher selectivity, sensitivity and accuracy of biological material analysis for tetramethylthiuramdisulfide.
4 ex, 5 tbl
SUBSTANCE: conjunctival cell sample is collected by pressing a bulbar conjunctiva of an examined eyeball at 2-3 mm above a limb in the meridian 12 hours with a soft contact lens (SCL) placed with its concave side on a tonometre. Then, the SCL is removed from the tonometre and turned out so that the collected sample is at the bottom of the SCL which is fixed in 95% alcohol by single dip. Then, the SCL is washed with distilled water, then coloured with hematoxylin for 10-15 seconds, wash with distilled water and air-dried. Thereafter, the SCL is placed on a slide surface for cytological analysis.
EFFECT: invention allows simplifying the method for conjunctival cell preparation for cytological analysis, reduced analysis time and cost.
SUBSTANCE: heparinised blood, isotonic solution containing 0.1 % nitroblue tetrazolium are introduced in plate wells. The mixture is incubated for 30 minutes at 37°C, added with 3% acetic acids, heparinised blood preliminary dissolved in isotonic solution of BCG vaccine containing 0.1 % nitroblue tetrazolium. Further, the prepared mixture is incubated for 30 minutes at 37°C, added with 3 % acetic acids. It is followed with counting neutrophils with observed cytoplasm violet-blue formazan granule depositions in a Gorjaev's chamber with taking into account 100-200 neutrophils, and the percentage of positive NBT-cells is assessed.
EFFECT: use of the method enables higher accuracy and reliability of determining neutrophil capacity and reduced acquisition time.
3 tbl, 3 ex
FIELD: veterinary science.
SUBSTANCE: artificial gastric acid is composed according to prescription: distilled water of temperature 41-42°C - 1000.0 ml, concentrated hydrochloric acid with specific weight of 1.2 - 12.0-15.0 ml, porcine food pepsin 20-30 g. Incubation is carried out in "Gastros" device at the temperature of 41-42°C. Broth is settled, afterwards the residue is centrifuged. Then 8.0 ml of upper fluid level is aspirated from test tube, a drop is taken from lower level, and a smear is made on slide plate. Remaining part is applied with a dropper onto slide plates and dried, then made preparations are stained according to Romanovskiy-Giemsa for 10 minutes and are investigated under microscope with increase of ×400 and ×1250 with immersion.
EFFECT: method is convenient and simple to use, makes it possible to validly detect sarcocyst trophozoites.
SUBSTANCE: for an assay, 5-7 cm3 of blood is taken, and before extraction the sample is pre-treated with 2 cm3 of 60% sulphuric acid; the extraction process is executed with 30 cm3 of n-hexane once, and gas chromatography is preceded with single treatment of n-hexane extract with 10 cm3 of concentrated sulphuric acid.
EFFECT: invention provides higher reliability of α-HCCH, γ-HCCH test results and twofold reduced time of sample preparation.
1 ex, 1 tbl
SUBSTANCE: cystic bile recovered by fractional duodenal intubation is analysed for the concentration of cholic acid, phospholipids, cholesterol, bilirubin with calculating cholate- cholesterol (CCC) and phospholipid- cholesterol (FCC) coefficients. If observing decreasing cholic acid to 14.13±2.54 mmol/l, phospholipids to 1.62±0.27 mmol/l, bilirubin to 1.94±0.24 mmol/l, and increasing cholesterol to 9.04+1.35 mmol/l, decreasing the CCC to 1.59±0.22 and the FCC to 0.18+0.027, more severe clinical form of psoriasis, mainly erythrodermatitis developing is predicted.
EFFECT: invention provides exact prediction of transition of plaque psoriasis accompanied with chronic acalculous cholecystitis to erythrodermatitis.
4 tbl, 3 ex
SUBSTANCE: diagnostic technique for latent radiation sickness consisting in biopsy procedure of externally unaffected leg skin followed with histological study of a biopsy material, and if certain morphological signs of inflammatory changes of microvasculature vessels presented, latent radiation sickness is diagnosed.
EFFECT: technique allows diagnosing radiation sickness at early stages of development.
1 tbl, 1 ex
SUBSTANCE: method of early prediction of septic complications in newborns with a respiratory pathology, by examining venous blood by direct immunofluorescence on a flow cytometre, a relative monocyte Annexine V concentration is evaluated, and if observing the value equal to 17.4 % and more, septic complications are diagnosed.
EFFECT: method exhibits high sensitivity and specificity and refers to near-patient testing.
SUBSTANCE: before and after noon, urine samples are collected for 5-6 hours. Thereafter, two native urine samples are collected, the second sample is mixed with 10% albumin in the ratio 4:1 respectively. The mixture is dropped on a transparent surface, dried to carry out microscopic analysis, and if observing in the middle of the native urine sample isotropic salt X-crystals and isotropic salt crystals in the mixture sample, chronic renal insufficiency is diagnosed.
EFFECT: use of the technique allows fast and demonstrative detection of insufficient renal function in screening examination of patients.
2 ex, 5 dwg
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.