Laboratory diagnostic technique for sepsis
SUBSTANCE: substance of the laboratory diagnostic technique for sepsis consists in a quantitative analysis of blood serum for nonprotein microorganism metabolism products, particularly n-hydroxyphenyllactic (HPLA), phenyllactic (PLA), n-hydroxyphenylacetic (HPAA) and homovanillic (HVA) acids. If the contents of said four acids listed above synchronously excess the a healthy level by 5 times or more, sepsis is diagnosed.
EFFECT: use of said technique allows higher reliability of diagnosing sepsis in the patients with hyperthermia of an uncertain aetiology, in impaired state of the patients in hopital, in the intensive care patients, and in surgical postoperative complications.
4 tbl, 3 ex
The method can be used in clinical practice for diagnosis of sepsis (generalized bacterial infection) in patients with pyrexia of unknown etiology, when the deterioration of the patients in the hospital, patients in the intensive care unit and postoperative complications in surgery for the differential diagnosis of sepsis from other diseases and conditions.
The invention relates to the field of clinical laboratory diagnostics and can be used in clinical practice, the analysis of whole blood for the diagnosis and differential diagnosis of generalized bacterial infection (sepsis).
Today the problem for the laboratory diagnosis of sepsis is not solved. In accordance with international norms, the diagnosis of sepsis is common to put together clinical and laboratory parameters, including the presence of purulent or bacteremia in combination with signs of the syndrome of systemic inflammatory response, namely with tachycardia, tachypnea, hyperthermia, leukocytosis and shift leukocyte formula to the immature forms of neutrophils (American College of Chest Physicians/Society of Critical Care Medicine Consensus Conference. Definitions for sepsis and organ failure and guidelines for the use of innovative therapies in sepsis Chest 1992; 101: 1644-55). Diagnosis of sepsis represents a major challenge for clinicians, taktak the above symptoms are not specific and are found in a variety of diseases and conditions.
Search laboratory markers of sepsis is mainly in the study of macromolecular structures of polypeptide nature (Pro - and anti-inflammatory interleukins, tumour necrotic factor and others). Recently, as a laboratory criterion most often recommended test for procalcitonin - Oligopeptide, a precursor of the hormone calcitonin, increasing the content of which in the blood serum, 2 ng/ml correlates with bacterial load characteristic of septic conditions (Assicot M, Gendrel d, Carsin h, Raymond J., Guilbad j, Bohuon C. High serum procalcitonin concentrations in patienys with sepsis and infection. Lancet 1993 Vol.341 P.515-518). But still the nature of the increase in the content of procalcitonin in the blood of patients is not clear, moreover, elevated levels of procalcitonin observed in patients with severe multi-organ failure, out of touch with sepsis. For the diagnosis of anaerobic purulent-inflammatory processes use the definition of low molecular weight compounds representing metabolites of anaerobic bacteria, such as volatile fatty acids (AGV-propionic, butyric, valeric, Caproic and their isomers), however, the test AGV is not included in the set of techniques for the diagnosis of sepsis, as the latter is often associated with aerobic microflora.
One of the laboratory criteria are generalized and the burden, caused by gram-negative bacteria, is the high level of lipopolysaccharide (LPS) in the serum. This test is used in the scientific-experimental studies, because of its diagnostic significance in the clinic is not confirmed. Sepsis in most cases is a mixed infection caused not only gram-negative and gram-positive microorganisms.
The proposed method provides as markers of sepsis to use low molecular weight chemical compounds, vinylcarbazole acid, which is directly by the metabolic products of bacteria, both gram-negative and gram-positive. (1. ..Jenner, J.Rafter, .Halliwell Human fecal water content of phenolics: The extent of colonic exposure to aromatic compounds Free Radical Biology & Medicine 2005, V.38, P.763-772; 2. A.R.Rechner, .A.Smith, G.Kuhnle, G.R.Gibson, E.S.Debnam, S.K.S.Srai, K.P.Moore, C.A.Rice-Evans Colonic metabolism of dietary polyphenols: Influence of structure on microbial fermentation products Free Radical Biology & Medicine 2004, V.36, No.2, P.212-225; 3. H.Ch.Lee, A..Jenner, Ch.S.Lowa, Y.K.Lee, Effect of tea phenolics and their aromatic fecal bacterial metabolites on intestinal microbiota Research in Microbiology 2006, V.157, P.876-884).
Closest to the proposed method is the determination of microbial markers in a sterile biological environments, based on gas chromatography-mass spectrometric detection of chemical substances which are structural components of microbial cells (Patent RF №2146368 Georgy Osipov on the Nikolaevich, Beloborodov Natalia Vladimirovna. The method of identifying the causative agent of an infectious process in a sterile biological environments of the microorganism, application No. 97117426 date 21.10.1997, priority 21.10.1997, registered in the State register 10.03.2000).
The disadvantage of the described method is that the estimation of microbial load on the basis of quantitative determination of a large list of substances contained in the cell walls of microorganisms (branched fatty acids, isocyclic, gidrokshikislota, alcohols, phenols, etc). Summing not only chemical compounds of living organisms directly involved in the septic process, but also structural components phagocytosing, lost activity, the dead and even completely destroyed bacteria.
The inventive method is characterized by the presence of features that distinguish it from known method of determining microbial markers in blood that is measured levels of metabolites, i.e. waste products of living organisms.
The method can be implemented using standard laboratory equipment with personnel qualification which does not exceed the level of a specialist in the field of chemical, biochemical, enzyme immunoassay.
For the diagnosis of sepsis is produced in the usual quantitative determination phenylcarbinol acids, namely, n-hydroxyphenylglycol (GFMC), phenylmalonic (PMK), n-hydroxyphenylglycol (GFWC) HVA (GVK) acids in human blood serum samples. Sepsis concentrations of these compounds are significantly higher than the background concentration (in healthy people) and the concentration of other diseases and conditions.
In contrast to classical methods of microbiological diagnostics (blood cultures), the proposed method does not require culturing bacteria on nutrient media, which significantly reduces the time of diagnosis. Raw material analysis is the serum of venous blood, which undergoes the process of sample preparation (dilution, acidification, the introduction of an internal standard, extraction in diethyl ether, evaporation, sicilianu) followed by gas chromatography-mass spectral analysis trimethylsilyl derivatives phenylcarbinol acids.
Method for quantitative determination of the content of phenylcarbinol acid serum
a) preparation of the internal standard solution.
16 mg of anhydrous D5benzoic acid is dissolved in 1 ml of 96% ethyl alcohol. 10 μl of the obtained solution was diluted with 96% ethanol to a volume of 4 ml of the contents of the D5benzoic acid in the final solution is 40 ng/ml or 40 μg/ml In the sample preparation process in the sample injected 10 m is l final solution (the absolute content of the standard 400 ng).
b) sample preparation
2 ml of whole venous blood centrifuged for 15 minutes at a rotor speed centrifuge 3000 rpm Select two parallel sample of 0.2 ml of the thus obtained serum, which are then processed independently and simultaneously. The serum is placed in a test tube with a capacity of 10 ml, diluted with 0.8 ml of water for injection, add 20 ál of 50% sulphuric acid (to pH 2) and 10 μl of an alcohol solution containing 400 ng D5-benzoic acid (internal standard). Then the resulting mixture was extracted twice with portions of 1 ml of diethyl ether. Received the ether extracts are combined and evaporated at 40°C to dryness. The dry residue is treated with 20 µl of N,O-bis(trimethylsilyl)trifurcated (BSTFA) for 15 minutes at 80°C in a confined space (in a tightly closed container). Received sililirovany product (TMS-derived) was diluted with 80 μl of n-hexane (qualification "OSC"), is transferred into the insert in the vial for holding chromatography-mass spektralnogo analysis (GC-MS).
C) GC-MS analysis. Gas chromatography-mass spectral determination forsteriana carboxylic acids is performed using a gas chromatograph Agilent Technologies 6890, equipped with mass spectral detector Agilent Technologies 5973. Khromatograficheskoe the separation of components prodat quartz HP5MS capillary column diametral,2 mm, with a length of 25 m, a thickness of 0.33 μm.
Chromatograph conditions were as follows:
the carrier gas helium, flow rate of 24 ml/min, the flow rate through the column - 1.2 ml/min; evaporator temperature of 280°C; the operating mode of thermostat - programmable: the initial temperature of the column thermostat to 80°C., the holding time is 4 min, followed by heating to 240°C at 7°/min to 320°C at 15°/min, then the temperature at 320°C until the end of the analysis; size of the sample - 2 ál, analysis time - 35 min, time delay detector 4 minutes operation mode of the detector - the full scan mode.
Identification forsteriana acids produced in accordance with the retention times of the primary and confirmation ion (m/z), are given in table 1.
|The retention time, intensity of the primary and confirmation ion (m/z) in the chromatograms and mass spectra forsteriana carboxylic acids|
|No.||Substance||The hold time. TMS-derivative, min||Est. ion, m/z (int. Rel. unit)||ACK. ion, m/z|
|1||D5Benzoic Ki the lot (standard)||10.45||184 (999)||110|
|2||n-Hydroxyphenylglycine acid||22.02||179 (999)||147|
|3||Phenylmalonate acid||17.12||193 (999)||147|
|4||n-Hydroxyphenylarsonic acid||18.04||179 (256)||296|
|5||HVA acid||20.05||209 (338)||326|
g) processing results
For the calculations using the chromatogram obtained in the current mode selective ions (SIM).
Spend the identification of peaks corresponding to the detectable substances in the retention times (table 1). The presence of a substance is proved in the case of presence in the chromatogram peaks for the primary and confirmation ion (m/z) with relative intensities close to him in the mass spectrum from the database NIST-02. Integrating (definition of area) peaks conduct psignal primary ion (m/z).
Mass concentration (ng/ml) forsteriana acids calculated by the method of internal standard according to the formulawhere cithe concentration of the i-th component (ng/ml), Sithe peak area of the i-th component defined for the primary ion (m/z), Mrimolecular weight of the i-th component of mstweight input standard (400 ng), Imax- height of the peak is the most intense ion (m/z) in the mass spectrum of TMS-derivative of the i-th component (999), Sstthe peak area of the standard, Mrstmolecular weight standard (127), Vs- sample volume (0.2 ml), Ii- - relative intensity (height) of the peak of the primary ion (m/z) in the mass spectrum of TMS-derivative of the i-th component. Mass spectral data (m/z) and relative intensity of peaks) taken from the international database of mass spectra of NIST-02.
The detailed survey of the contents in the serum GMPC, PMK, GFWC, GVK sepsis and control groups: in healthy individuals, in patients with heart disease non-infectious nature, with a local infectious-inflammatory processes (pneumonia). It is established that the levels phenylcarbinol acids in the serum of sepsis are many times greater than levels in all comparison groups. So, the content of GMPC, PMK, GFWC, GVK in the serum of patients with sepsis was not less than 5 times higher than the healthy, who's people.
Example 1. Investigated the content of microbial metabolites in the serum of 76 patients belonging to the 4 clinical groups: healthy donors (16), patients with cardiovascular disease (36 patients), patients with a diagnosis of pneumonia (10 people) and patients with a diagnosis of sepsis (14 people). Conducted full cycle of sample preparation and quantification phenylcarbinol acids. The obtained data statistically processed and presented in table 2.
|Medians and interquartile swings content phenylcarbinol acids (ng/ml) in the serum of sepsis and comparison groups|
|Donors||Patients with cardiovascular diseases||Patients with a diagnosis of pneumonia||Patients with a diagnosis of sepsis|
|The interquartile swing 75%||371||373||836||2505|
|The interquartile scale 25%||37||37||89||206|
|The interquartile swing 75%||64||93||177||849|
|The interquartile scale 25%||62||53||344||541|
|The interquartile swing 75%||220||667||2694|
|The interquartile scale 25%||3||13||357||356|
|The interquartile swing 75%||10||118||3493||2749|
The data obtained confirm the existence of significant differences in the levels of phenylcarbinol acids (GFMC, PMK, GFWC, GVK) in patients with sepsis compared with healthy people and patients with other diseases.
Example 2. Patient K., 42, East. bol. No. 54314/08, was admitted to the hospital 27.08.2008. Main diagnosis: rheumatic fever, mitral stenosis and insufficiency, comorbidities - diabetes, atrophic gastritis, the operation 27.08.2008, in the postoperative period developed hyperthermia, leukocytosis, bacteremia, pneumonia, sepsis, multiple organ failure. This patient with documented sepsis developed in the postoperative period, the administration is prohibited research content phenylcarbinol acids in the serum over time. The data are given in table 3.
|The dynamics of content phenylcarbinol acids (ng/ml) in the serum of the patient K. with documented sepsis|
Monitoring phenylcarbinol acids demonstrated a strong increase of their content in all samples of blood serum within one month the patient with sepsis. Was relatively small fluctuations in the background changing the overall condition and treatment, and higher concentrations were recorded at the time of the rise of clinical and laboratory signs of sepsis. The highest concentration GFMC, PMK and GFWC, GVK marked 13.10.08 when the patient was already in a terminal condition.
Example 3. To compare the diagnostic value of the proposed method and the method-prototype study the content of the microbial metabolites (GFMC, PMK, GFWC, GVK) and components of cell walls of bacteria (3h14, i14, A19) in patients with sepsis (n=10) and healthy donors (n=11). The research results are statistically processed and presented in table 4.
|Medians and interquartile swings content phenylcarbinol acids and components of cell walls of bacteria (ng/ml) patients belonging to different groups|
|Donors||Patients with a diagnosis of sepsis|
|The interquartile scale 25%||182||840|
|The interquartile swing 75%||505||4370|
|The interquartile scale 25%||42||325|
|The interquartile swing 75%||107||1182|
|The interquartile scale 25%||6||187|
|The interquartile swing 75%||120||2119|
|The interquartile scale 25%||2||853|
|The interquartile swing 75%||26||1932|
|The interquartile scale 25%||8||15|
|The interquartile swing 75%||15||33|
|The interquartile scale 25%||44||73|
|The interquartile swing 75%||107||126|
|The interquartile scale 25%||223||215|
|The interquartile swing 75%||320||575|
|* 3h14 - 3-hydroxydecanoate acid|
|** i14 -every-tetradecanoate acid|
|*** A19 - anties-nonadecanoic acid|
Comparative analysis of statistical data in the group of donors and in patients with a diagnosis of sepsis, obtained in the study of blood serum of the same patients by two methods shows that differences in the level phenylcarbinol acids (GFMC, PMK, GFWC, GVK) between groups is significantly higher than the level of components of cell walls of bacteria (3h14, i14, A19). So the m way the proposed method significantly improves the accuracy of diagnosis of sepsis in comparison with the prototype.
Method for the laboratory diagnosis of sepsis, including chemical analysis of the blood serum, the measurement of the concentration of low molecular weight compounds nonprotein nature as markers of sepsis, characterized in that as markers of sepsis use vinylcarbazole acid, namely n-hydroxyphenylglycol (GFMC), phenylalanyl (PMK), n-hydroxyphenylglycol (GFWC), HVA (GVK) acid levels in sepsis is not less than 5 times higher than the levels in healthy people.
SUBSTANCE: initial (3 to 12 day) assessment of clinical symptoms of disease (fever and exanthema) is combined with measuring blood serum C-reactive protein (CRP), erythrocyte sedimentation rate (ESR) and specific antibodies (AT) and if observing: - fever response for less than 2 days, exanthema duration for less than 1.5 days, the CRP value less than 7.5 mg/l, ESR less than 15 mm/hour; the IFN-γ level less than 75 pg/ml, circulating immune complex 0.115 optical density units and lower, and specific antibody production more than 8.32 log2, a chronic clinical course of yersiniosis is predicted; - fever response for more than 12 days, exanthema duration for more than 3 days, the CRP value more than 15 mg/l, ESR more than 30 mm/hour; the IFN-γ level more than 150 pg/ml, circulating immune complex 0.130 optical density units and higher, specific antibody production less than 7.32 log2, a severe yersiniosis with associating organ lesions is predicted; - fever response for more than 12 days, exanthema duration for more than 9 days, the CRP value more than 5 mg/l, ESR more than 50 mm/hour; the IFN-γ level less than 150 pg/ml, circulating immune complex 0.250 optical density units and higher, and specific antibody production more than 9.32 log2, clinical outcome of yersiniosis in a systemic disease is predicted.
EFFECT: prevention of a chronic and prolonged clinical course of the disease.
3 tbl, 3 ex
SUBSTANCE: invention relates to medicine, namely to oncology and can be used for predicting efficiency of local chemical therapy of brain in patients with malignant tumours of central nervous system. For this purpose during surgical intervention for ablation of brain tumour from perifocal zone of tumour taken is brain tissue, which is divided into samples, 100 mg each. Number of samples is limited by number of studied chemical preparations. Chemical preparationbs are added to samples in dose 0.2 mg. Incubation at temperature 37°C for 30 minutes is performed. Activity of alpha-2-macroglobulin is determined before and after incubation. If index increases by 40% and more, in comparison with the value before incubation, efficiency of chemical preparations is predicted.
EFFECT: method ensures high specificity, possibility of objective evaluation of efficiency of chemical preparation impact in performing analysis on operation day, which makes it possible to start adequate therapeutic measures, including cancelling or replacement of chemical preparation in each specific case.
SUBSTANCE: diagnosing of early congenital latent syphilis is enabled by determining the values of acute cell-mediated, humoral components of the immune system and phagocytosis. If the CD19 +-lymphocyte percentage values exceed 12.1 as related to the reference group, the absolute and relative HLA-DR-antigen number values: more than 80.6×103/mcl and 1.5 % respectively, than in the reference group, and also a phagocytic neutrophil index is more than 48.2 %, latent ECS is diagnosed.
EFFECT: use of the method faster and more accurate diagnosing of latent ECS.
2 tbl, 2 ex
SUBSTANCE: test device has two surface impermeable layers, a layer of absorbing matrix material on which the reagent is applied and which is placed between the impermeable layers. One of the impermeable layers has multiple openings superimposed with the matrix material and capable of absorbing the sample into the matrix material from the surface. The device is laminated and is fitted with apparatus for separating the sample from the surface when rubbed. Disclosed is an alternative version of the test device, versions of the method of extracting the sample from the surface of test devices and a method of making the test device. The invention increases adhesion strength between layers of the test devices and ensures separation of the sample from the surface of the devices when rubbed.
EFFECT: high-speed and large-scale manufacturing of test devices.
58 cl, 6 dwg
SUBSTANCE: reagent for testing total α-amylase activity in serum, blood plasma and urine contains as a substratum 2-chlorine-4-nitrophenl-4-O -β-D-galactopyranosylmaltoside (GalG2CNP), sodium chloride, potassium sulphocyanate, sodium azide, EDTA, calcium acetate, Triton X-100, 2-morpholinoethanesulfonic acid (MES) and water. The pH value of said reagent is 5.5-6.5.
EFFECT: use of the reagent enables high precision and results reproducibility α-amylase test.
1 tbl, 4 ex
SUBSTANCE: invention concerns a kit of reagents for pancreatic α-amylase activity test including a reagent 1 containing sodium chloride, potassium sulphocyanate, sodium azide, water-soluble calcium salt, monoclonal salivary α-amylase antibodies (MAB), 2-morpholinoethanesulfonic acid (MES) and water, and a reagent 2 containing 2-chloring-4-nitrophenyl-4-O-β-D-galactopyranosylmaltoside (GalG2CNP), MES and water, differing by the fact that the reagent 1 in addition contains EDTA and bovine serum albumin (BSA), and as calcium salt, it contains calcium acetate, and the reagent 2 in addition contains EDTA and sodium azide in the proportions specified in the patent claim.
EFFECT: enhanced stability of the set of reagents with maintaining high accuracy and result reproducibility of the analysis.
1 tbl, 4 ex
SUBSTANCE: invention relates to field of medicine, namely to methods of predicting post-operational complications, namely to methods of predicting development of scars after previous acne. In order to predict scar development content of receptor antagonist of interleukin-1 (RAIL) is determined during 15 days after resolution of inflammatory process. Scarless development of process is diagnosed at level RAIL in peripheral blood serum after disease being within physiological norm (300-800 pg/ml). If level of RAIL is lower than said norm prediction of acne complication in form of skin scars is diagnosed. Possibility of development of hypertrophic scars is predicted if RAIL concentration is lower than 200 pg/ml.
EFFECT: method makes it possible to predict type of complications after previous acne, therefore correcting therapy carried out in due time can prevent risk of skin scar formation and improve patient's life quality.
2 cl, 1 tbl, 4 ex
SUBSTANCE: developing hypoxia in a pregnant woman of the third trimester of gestation is predicted by evaluating peripheral blood oxyhemoglobin (HbO2) and 2,3 diphosphoglycerate phosphatase (2,3DPG) concentrations. A discriminator (D) is calculated by formula D=+17.072·2.3DPG + (-0.041·HbO2), 2,3DPG - concentration, mol/l, HbO2 - amount, %. The D value within 97.09-112.37 enables to predict threatened hypoxia in a pregnant woman had an acute condition of herpes virus infection.
EFFECT: use of the method allows well-timed detection of a risk group and correct prediction of developing hypoxia.
SUBSTANCE: threatened reduced erythrocyte oxygenation in a pregnant woman suffering an acute attack of bronchial asthma in the first trimester of gestation is predicted by evaluating oxyhemoglobin (HbO2) and 2,3 DPG (2,3 diphosphoglycerate phosphatase). This is followed by calculating a D discriminator value by formula D=18.05·2,3DPG + (-0.075·HbO2), and observing the D value within 88.66 to 102.42, threatened reduced erythrocyte oxygenation that leads to hypoxia is predicted.
EFFECT: use of the method enables predicting threatened reduced erythrocyte oxygenation in a pregnant woman with the acute attack of bronchial asthma.
SUBSTANCE: differentiated detection large and small circulating immune complexes in blood serum is ensured by a follow-on examination of blood serum clarified by short centrifugation with a method of circulating immune complexes precipitation PEG -6000 of the end concentration 4 % and 6 %. Thereafter, the results are recorded by a turbidimetric method with using a microplate spectrophotometer in a two-wave mode: basic - 340 nm, reference filter - 620 nm. Duration of an incubation step at temperature +18-25°C is 15 minutes. Using the method enables higher effectiveness and reliability of determining the level of large circulating immune complexes, results reproducibility, as well as differential measurement of the level of small CIC which are a diagnostically significant indicator of human body immune responsiveness in many types of a pathology, decreased volume of analysed serums to 0.06 ml, and incubation duration to 15 minutes.
EFFECT: method is suitable for clinical screenings.
1 dwg, 1 tbl, 1 ex
SUBSTANCE: invention describes a method of quantitative evaluation of blood acetic, propionic, isobutyric, butyric, valeric, isocapronic and capronic acids by gas chromatography analysis wherein a blood sample is acidified with 1 % sulphuric acid to pH 2-3, evaluated acids are extracted with isobutyl alcohol volume of which is related to the blood sample volume as 1:1. The protein separation is enabled by centrifugation. 2-3 drops of 0.4 % alkali is added, and the extract is evaporated dry, further the solid residue is added consistently with 1 % sulphuric acid and isobutyl alcohol that is followed with gas chromatography separation of the mixed acids in a capillary column with a flame ionisation detector, and the amount of each acid is evaluated by a calibration diagram.
EFFECT: higher sensitivity and accuracy of the method of quantitative evaluation of acetic, propionic, isobutyric, butyric, valeric, isocapronic and capronic acids if found in blood together.
5 cl, 1 ex, 4 tbl
SUBSTANCE: invention relates to field of medicine, namely, to gastroenterology. For non-invasive diagnostics of fibrosis and cirrhosis in case of HBV and HCV infections complex ultrasonic examination of liver and spleen tissue is carried out. Additionally duplex scanning with colour Doppler mapping of porto-hepatic region vessels is performed and quantitative indices of hemodynamics of rate of blood flow in vessels, including splenic vein, are determined. Blood test is analysed and used to determine number of platelets, biochemical blood tests are performed, most significant for determination of disease degree indices of coagulogram are taken. After that, discriminant analysis of obtained characteristics and indices is carried out, and taking into account age and experimentally obtained coefficients, total value of two canonical discriminant functions F1 and F2 for HCV and HBV is calculated. Further, by obtained in empiric way territory map position of point for calculated by patient's concrete indices values F1 and F2 for cases of HCV-infection and HBV-infection is determined. Depending on point location on territory map case of mild fibrosis, severe fibrosis or liver cirrhosis is diagnosed.
EFFECT: method increases reliability of fibrosis and cirrhosis diagnostics in case of HBV and HCV infections.
10 dwg, 2 ex
SUBSTANCE: peripheral blood thrombocytes of women suffering gestosis of various severity levels on their 32-38 weeks of pregnancy are analysed for the activity of glutathione reductase (GY), NADF-dependent glutamate dehydrogenase (NADFGDG) and NADF-dependent isocitrate dehydrogenase (NADFICDG). A nicotine amide adenine dinucleotide phosphate transfer coefficient (NTC) represented by the relation of the GY activity to a product of the NADFGDG and NADFICDG activities is calculated. At the NTC value is equal to 1.3 and lower, the newborn's Apgar score is predicted to be equal to 6 and less, and the NTC value exceeding 1.3 provides the Apgar score being 7-10 points.
EFFECT: more accurate prediction of the newborn's state.
2 tbl, 5 ex
SUBSTANCE: areas highly exposed to harmful chemical agents are chosen. A random group of children without clinical signs of living in this territory is tested using chemical laboratory tests of blood to identify the content of chemical compounds, which are priority chemical environmental factors on the selected area of residence and clinical and laboratory studies are conducted to determine a set of laboratory indicators of adaptation system. Then using the results of the study the average values of chemical compounds in the blood are fixed and then they are compared to the background and average values for each of the above laboratory parameters are fixed and compared to the physiological norm; deviation of this value from the normal rate reveals children bodies response to chemical exposure. Next, a causal relationship is established between the level of content of chemical compound in the blood and the response of the child body through deviation of laboratory parametres from the norm using a logistic regression model. Using method based on analysis of odds ratios the maximally inactive level of marker of exposure and corresponding response marker based on the conditions are determined under which the odds ratio that characterises the degree of the connection between exposure to a chemical compound and the body's response will be greater than or equal to one; for this a model of dependence between the level of a marker of exposure and the specified index odds ratio is designed, the parametres of the model are determined and they reflect the change in the probability using which the value of the maximally inactive level of marker of exposure is calculated, i.e. maximum ineffective concentrations of chemical compounds. From the entire spectrum of defined concentrations of certain chemical compounds for each laboratory parametre of adaptation systems choose the smallest value that is accepted as the maximally inactive concentrations on a child adaptation system for a given chemical compound i. In future diagnosis of violation of adaptation of children living in the selected area is performed by comparing the content Ci of certain chemicals in their blood with previously established value of inactive concentration for this chemical compound; and if a ratio is a violation of adaptation is diagnosed.
EFFECT: method allows diagnosing violation of children adaptation under chemical hazards of environmental factors with high precision at an early preclinical stage, with simultaneous simplicity and accessibility for a wide practical application.
5 tbl, 2 dwg
SUBSTANCE: zeolite antioxidant activity test is enabled by introducing a substance being tested into bodies of experimental animals. Biological products of tissues and organs of the experimental animals and control sets are prepared. Metabolic process indicator substances are evaluated. The pulmonary tissue, blood plasma, erythrocytes and thrombocytes are analysed for the content of lipid peroxidation products and natural antioxidants which are scored and summed up. The zeolite antioxidant activity is tested relatively to the normal values of the content of lipid peroxidation products and natural antioxidants which are defined as an arithmetical mean of the relevant values received in the animals of a control set.
EFFECT: enabled reliable zeolite antioxidant activity test at the enabled comparative evaluation of substances by this parametre.
2 tbl, 1 ex
SUBSTANCE: vaginal fluid is analysed. The vaginal secretion is collected by means of a common tampon placed in a vagina for 8-9 hours, further weighted; microbial metabolites are extracted in equiponderate amount of distilled water, and the extract is analysed by gas-liquid chromatography. If the vaginal discharge contain acetic acid more than 0.315 mg/g and total propionic and butyric acids ≤0.200 mg/g in an age group of 17 to 34 years, and acetic acid more than 0.210 mg/g with total propionic and butyric acids ≤0.120 mg/g in an age group of 35 to 48 years, nonspecific aerobic vaginitis is diagnosed.
EFFECT: more accurate diagnosis of nonspecific aerobic vaginitis.
SUBSTANCE: method of evaluating immunogenicity of brucella strains includes enzyme-linked analysis of culture supernatant of peripheral blood cells for content of cytokines - tumour necrosis factor (TNF-α), interleukin-1β (IL-1β) and colony-stimulating factor (CSF), synthesized by mononuclear cells of peripheral blood in vitro without impact (spontaneous production) and under the impact of antigens of evaluated brucella strains (induced production) and determination of their immunogenicity by ratio of spontaneous and induced production of said cytokines, brucella strains are considered immunogenic if their antigens cause enhancing of TNF-α by 1-1.25, IL-1β by 2-2.50 and CSF by 3-3.70.
EFFECT: method improvement.
SUBSTANCE: sampling of patient's lacrimal fluid (LF) is performed, analysed reaction mixture (ARM), consisting of substrate and analysed lacrimal fluid (ALF) is prepared, where as substrate, collagen gel is used. In course of ARM preparation, ALF is mixed with collagen gel in ratio 1-1.5:1 and keep at room temperature until homogeneous mixture (said ARM) is obtained. After that said ARM is applied on a microscope slide, kept until complete drying up of the entire microscope slide surface, measuring the time of complete ARM drying up. Then, quantitative determination of ALF CA is carried out by means of preliminarily built calibration curve of dependence of time of complete drying up of standardised reaction mixture samples, each of which consists of substrate and collalisin solution with specified collagenolytic activity, on collagenolytic activity. Obtained earlier ALF CA value is compared with normal values and if value of collagenolytic activity is lower than 231.8 kU/ml, lower ALF CA is determined, if the value of said activity is 231.8-297.8 kU/ml, normal ALF CA is determined, and if the value of said activity is higher than 297.8 kU/ml, higher ALF CA is determined.
EFFECT: application of the method makes it possible to increase accuracy of LF CA determination, reduce duration of determination procedure, eliminate possibility of infecting people who are carrying out the analysis.
1 dwg, 3 tbl
SUBSTANCE: invention refers to medicine, namely to oncology, and can be used for clinical effectiveness control in children with neuroblastomas. That is ensured by neo-adjuvant combination cytostatic therapy with peripheral vein blood sampling prior to and after each course of chemotherapy. Blood is examined for plasminogen and plasmin activity to calculate the relation of the first to the second. If the value increased after chemotherapy, a therapeutic clinical effect is predicted. If the value decreased or remained unchanged throughout two courses of chemotherapy, the absence of effect is predicted that is a basis for changing the cytostatics to provide an adequate treatment.
EFFECT: method provides assessing cancer invasiveness, its invasive and metastatic potential, detecting the patients with an expected therapeutic effect, good prognosis and the patients with no effect who require timely correction of anticancer therapy to provide prolonged and improved quality of life of the patients.
2 ex, 1 tbl
SUBSTANCE: invention concerns a method for prediction of the efficacy of Infliximab inclusion in a conventional therapy of the patients with rheumatoid arthritis (RA). A patient's peripheral blood is examined for total lymphocyte content (TLC) expressing TNF-α-CD120a receptors, for levels of spontaneous and stimulated production of tumour necrosis factor alpha (TNF-α) and interleukin-6 (IL-6). It is followed by calculating indexes of PHA action on PBMC (peripheral blood mononuclear cells) TNF-α and IL-6 production by formulae respectively: IA1=Ast/Asp, IA2=Bst/Bsp, where IA1 is an index of PHA action on PBMC TNF-α production; Ast is a level of stimulated TNF-α production, pg/ml; Asp is a level of spontaneous TNF-α production, pg/ml; IA2 is an index of PHA action on PBMC IL-6 production; Bst is a level of stimulated IL-6 production, pg/ml; Bsp is a level of spontaneous IL-6 production, pg/ml. Provided IA1≤2.4 and IA2≤1.6 and TLC≥6% simultaneously, the high efficacy of Infliximab inclusion in the therapy of the RA patients is predicted.
EFFECT: invention allows reliable prediction of the efficacy of Infliximab inclusion in the conventional therapy of the RA patients.
FIELD: medicine, hepatology.
SUBSTANCE: one should detect the level of hepato-specific enzymes (HSE) in blood plasma, such as: urokinase (UK), histidase (HIS), fructose-1-phosphataldolase (F-1-P), serine dehydratase (L-SD), threonine dehydratase (L-TD) and products of lipid peroxidation (LP), such as: dienic conjugates (DC), malonic dialdehyde (MDA). Moreover, one should detect the state of inspecific immunity parameters, such as: immunoregulatory index (IRI) as the ratio of T-helpers and T-suppressors, circulating immune complexes (CIC). Additionally, one should evaluate the state of regional circulation by applying rheohepatography (RHG), the system of microhemocirculation with the help of conjunctival biomicroscopy (CB) to detect intravascular index (II). In case of increased UK, HIS levels up to 0.5 mcM/ml/h, F-1-P, L-SD, L-Td, LP products, CIC by 1.5 times, higher IRI up to 2 at the norm being 1.0-1.5, altered values of regional circulation, increased II up to 2 points at the norm being 1 point, not more one should diagnose light degree of process flow. At increased level of UK, HIS up to 0.75 mcM/ml/h, F-1-P, L-SD, L-TD, LP products, CIC by 1.5-2 times, increased IRI up to 2.5, altered values of regional circulation, increased II up to 3-4 points one should diagnose average degree of process flow. At increased level of UK, HIS being above 0.75 mcM/ml/h, F-1-P, L-SD, L-TD, LP products, CIC by 2 and more times, increased IRI being above 2.5, altered values of regional circulation, increased II up to 5 points and more one should diagnose severe degree of process flow.
EFFECT: higher accuracy of diagnostics.