Method of antioxidant activity assessment

FIELD: medicine.

SUBSTANCE: invention belongs to experimental and clinic medicine, notably to medical biophysics, clinic pharmacology, biochemistry, oncology and immunology. For antioxidant activity assessment, test specimen of an oxidant is injected into biological target, and then in target free-radical oxidation is induced. Antioxidant activity is assessed by optical density of the patterns within photometry. As biological target whole blood with addition of citrate or heparin is used. Test specimen of an oxidant is injected in 5% suspension made of whole blood, after incubation 10 mcl of 1 mM clotrimazole suspension is added. To induce oxidation 20 mcl of 100 mM tert-butyl hydroperoxide solution is added. Mix is once again incubated, centrifuged, and then supernatant's optical density is measured.

EFFECT: method allows increasing of accuracy and effectiveness of antioxidant activity assessment in natural and chemical compounds and biological liquids.

6 cl, 2 dwg

 

The invention relates to experimental and clinical medicine, in particular to medical Biophysics, clinical pharmacology, biochemistry, Oncology and immunology, and can be used for biomedical research with the aim of determining the antioxidant activity of different natural and chemical compounds, and biological fluids.

As is well known, the mechanisms of action of reactive oxygen species (ROS) are quite varied, complex, and to date not fully understood. Given the efficacy of antioxidant drugs in the correction of conditions associated with the action of oxidative stress, we have urgent task of assessing antioxidant activity (AOA) of such drugs.

To date, to assess the antioxidant activity of individual compounds, mixtures or total antioxidant activity of biological objects developed many methods. These methods differ in the type of oxidant and oxidizable compounds (targets), as well as the way of measuring the oxidized product. They provide a wide range of results that are not absolute values, and should be interpreted in light of the results obtained by other methods. Come to this conclusion the authors of numerous publications, both domestic and foreign. In particular, in various ways is usually used for testing are given in the section "review of the Methods for testing antioxidant activity" (Michael Antolovich et al., Journal of the Royal society of chemistry ANALYST, August 2002, .183-198).

As a prototype we have considered the method of evaluation of antioxidant activity of the sample (US 7132296, 2006), which describes the method for determining the index of ORAC (oxygen radical absorbance capacity), which today is a standard indicator of antioxidant activity in some countries, for example USA. The method consists in the fact that the antioxidant activity evaluated by introducing investigated the antioxidant to the mixture of sodium salt of fluorescein (F1) and 2,2'-Azobis(2-amidinopropane) dihydrochloride (AAAS). When incubation temperature of 37°C. carry out the measurement of the fluorescence intensity of the mixture. The intensity decreases as the oxidation F1 peroxyl radicals generated by thermal degradation of AAAS. The presence of antioxidants increases the period of time until a decrease in fluorescence intensity. The method requires fairly sophisticated equipment and costly in terms of reagents. In addition, conditions such analyses are far from physiological.

The technical result of the invention aims to remedy these shortcomings lies in the physiology of the measurement conditions, the ease of use and minimal consumption of reagents.

The essence of the proposed method lies in the fact that in the method of assessment of the key antioxidant activity by introduction of a test of an antioxidant in a biological object, induction of free radical oxidation and evaluation of antioxidant activity by the optical density of samples at their photometry as a biological object using whole blood, taken on the citrate or heparin, preincubated obtained from whole blood 5% suspension of erythrocytes with the investigated antioxidant, add a suspension of clotrimazole (final concentration 10 μm), after which induce oxidation, contributing to the solution of tributyl gidroperekisi (t-BHP) at a final concentration of 2 mm and incubated samples, then the samples are centrifuged and measure the optical density of nadeshiko.

The incubation is conducted for 3 hours at 37°C.

Preferably, the optical density of samples was measured at a wavelength of 490-540 nm.

Preferably the introduction of the investigated antioxidant in the suspension of erythrocytes in different concentrations, while the incubated samples 20 minutes After which the optical density of the samples measure the degree of hemolysis, build its dependence on the concentration of the investigated antioxidant and as an indicator of antioxidant activity using the antioxidant, leading to a 50% reduction in the degree of hemolysis.

Also as an indicator of antioxidant activity can be used the ratio of concentrations studied antioxi the Anta and the standard antioxidant trolox.

Previously proposed methods of estimating the AOA, which used the erythrocytes subjected to the action of azoinitiator (AAAS or AMVN) as sources peroxyl radicals (Kitagawa S, Sakamoto N, Tano N. Inhibitory effects of flavonoids on free radical-induced hemolysis and their oxidative effects on hemoglobin // Chem Pharm Bull (Tokyo). - 2004. - V.52, no. 8. - Pp.999-1001; Dai F, Miao Q, Zhou B, Yang L, Liu ZL. Protective effects of flavonols and their glycosides against free radical-induced oxidative hemolysis of red blood cells // Life Sci. - 2006. - V.78, No. 21. - Pp.2488-493; Abella A, Messaoudi C, Laurent D, Marot D, Chalas J, Breux J, Claise C, Lindenbaum A. A method for simultaneous determination of plasma and erythrocyte antioxidant status. Evaluation of the antioxidant activity of vitamin E in healthy volunteers // Br J Clin Pharmacol. - 1996. - V.42, №6. - Pp.737-741 and others).

It is shown that the presence of azoinitiators does not cause intracellular oxidation of hemoglobin and, accordingly, the subsequent release of hemin (Lenfant F, Bureau A, Lahet JJ, Bouyer F, Chaillot B, Freysz M. Effects of an oxidative stress on human hemoglobin: a multiwavelength visible spectrometry study.// Biomed Pharmacother.- 2005.- V.59, No. 5, Pp.230-232).

Meanwhile, the oxidation of hemoglobin and its subsequent degradation often occur in vivo in physiological (aging of erythrocytes)and pathological (malaria, ischemia - reperfusion injury, sickle cell anemia and other conditions. Thus, the red blood cells modified with oxidizing agents cause the formation of methemoglobin, including tributyl hydropredict (t-BHP), appear to be more adequate and biologically significant of cleoc the second model for the assessment of antioxidant activity in comparison with existing models, based on the use of azo-compounds as exogenous sources of free radicals.

It was shown that clotrimazole forms with exogenous gem (Eminem) complex, which has much greater political impact than the free heme (Huy NT, Kamei K, Yamamoto T, Kondo Y, Kanaori K, Takano R, Tajima K, Hara S. Clotrimazole binds to heme and enhances heme-dependent hemolysis: proposed antimalarial mechanism of clotrimazole // J Biol Chem. - 2002. -V.277, No. 6, Pp.4152-4158).

In the course of our work for the first time it was shown that clotrimazole at a concentration of 10 µm significantly enhances both the swelling and hemolysis of red blood cells under the action of t-BHP, which indicates a significant increase in damage to membranes in the presence of this compound. It is obvious that peroxide-induced hemolysis is, at least partly, the result of the lytic effect of hemin emitted from hemichrome formed in the cell during the oxidation of hemoglobin. The degree of hemolysis increases significantly if the suspension is added clotrimazole, which forms with Eminem complex, with more in comparison with the free Eminem lytic activity.

Thus, it is possible to consider clotrimazole as an agent enhancing oxidative hemolysis of erythrocytes under the action of t-BHP.

It is known that antioxidants reduce the degree of induced Eminem and t-BHP damage to the membrane and subsequent hemolysis./p>

Inhibitory effect of antioxidants undertaken at the initial stages of the process, further subjected to amplification in the presence of clotrimazole that probably explains the exceptionally high sensitivity of the proposed cell model to different antioxidants.

Figure 1 shows the scheme of carrying out of the method of estimating the AOA.

Figure 2 shows the dependence of the degree of hemolysis induced by t-BHP in the presence of clotrimazole, concentration of antioxidants.

The proposed method is as follows.

Whole blood taken on citrate or heparin, washed with saline (figure 1) and resuspending the washed red blood cells in HEPES-buffer containing 10 mm HEPES, 5 KCL, 0.8 MgSO4, 1.5 CaCl2, 5 glucose at pH 7.4, at a hematocrit of 5%.

70% aqueous solution of t-BHP was diluted with saline to a concentration of 100 mm.

Alcohol solution of clotrimazole 100 mm store in the refrigerator for 1-2 months and just before use, dilute it with saline to a concentration of 1 mm.

To 1 ml of 5% suspension of erythrocytes add studied the antioxidant in different concentrations, preincubated 20 minutes, then make 10 l suspension of clotrimazole, the final concentration of clotrimazole 10 μm, and then immediately make a 20 l 100 mm solution of tributyl gidroperekisi (BHP), the final concentration of 2 mm.

Prepared samples incubated for 3 hours in a thermostat at 37°C and continuous stirring. Then the samples are centrifuged in the Eppendorf centrifuge at 7000 rpm for 5 minutes

100 μl nadeshiko contribute in flat-bottomed wells containing 200 μl of a solution Drabkina.

Then measure the degree of hemolysis on the optical density of samples at λ=490 nm on a tablet spectrophotometric plate reader (spectrophotometer) and build curves degree of hemolysis on the concentration of the studied antioxidant. The antioxidant activity is expressed as the concentration of antioxidant corresponding to 50% inhibition of hemolysis (IC50). As a relative indicator of antioxidant activity using the ratio of the concentrations of the investigated antioxidant and standard antioxidant trolox (IC50/ICthe trolox50).

Figure 2 presents the dependence of the degree of hemolysis, expressed in % of the maximum (i.e. hemolysis observed without the presence of antioxidants). Suspension of erythrocytes was preincubated for 10 min with different concentrations of antioxidants, then added clotrimazole, 10 µm and tributyl Gidropress, 2 mm, and incubated for 3 h, n=5-20.

As the AOA indicator is used, the concentration of antioxidant, resulting in 50% snizeni the degree of hemolysis. Antioxidant activity can also be expressed as the ratio of the concentrations of the investigated antioxidant and standard antioxidant trolox.

Using the proposed method it is possible to determine the antioxidant activity of various compounds as hydrophilic and hydrophobic. We studied the effect of known flavonoids, as well as some tetracycline antibiotics, anti-oxidant which in recent years has been the subject of study of different groups of researchers.

The proposed method is highly sensitive, simple and easy to use, requires minimal amount of whole blood (~ 100 l) and used reagents.

Furthermore, the method allows the simultaneous screening of several compounds with the proposed antioxidant properties.

Also the proposed method is appropriate to use in the clinic, since the degree of hemolysis of erythrocytes under the influence of tributyl gidroperekisi from healthy donors may differ from patients suffering from certain diseases associated with the activation of free radical processes in the body. Thus, the degree of hemolysis of erythrocytes treated with tributyl hydropredict, is diagnostically or prognostically significant criterion or method of assessing the effectiveness of drug therapy.

1. Pic is b evaluation of antioxidant activity by introduction of a test of an antioxidant in a biological object, induction of free radical oxidation and evaluation of antioxidant activity by the optical density of samples at their photometry, characterized in that as a biological object using whole blood, taken on the citrate or heparin, administered analyzed antioxidant obtained from whole blood 5% suspension of red blood cells in HEPES-buffer, incubated contribute 10 l suspension of clotrimazole at a concentration of 10 mm and induce oxidation by adding 20 l solution of tributyl gidroperekisi at a concentration of 100 mm, re-incubated samples, then the samples are centrifuged and measure the optical density of nadeshiko.

2. The method according to claim 1, characterized in that the re-incubation carried out for 3 h at 37°C.

3. The method according to claim 1, characterized in that the optical density of nadeshiko measured at a wavelength of 490-540 nm.

4. The method according to any one of claims 1 to 3, characterized in that the antioxidant is introduced into a suspension of erythrocytes in different concentrations and incubated the samples for 20 min at room temperature.

5. The method according to claim 4, characterized in that the optical density of nadeshiko determine the degree of hemolysis, build its dependence on the concentration of the investigated antioxidant and as an indicator of antioxidant activity using the antioxidant, causing 50% reduced the th degree of hemolysis (IC 50).

6. The method according to any one of claims 1 to 4, characterized in that as a relative indicator of antioxidant activity using the ratio of the concentrations of the investigated antioxidant and standard antioxidant trolox (IC50/ICthe trolox50).



 

Same patents:

FIELD: medicine.

SUBSTANCE: invention belongs to medicine, notably to morphology and hystochemistry. In this method of histamine rate assessment in epithelial cells of rats endometrium within sex cycle, sample acquisition and following fluorescent-histochemical analysis with orthophtalic aldehide is made. In each phase of sex cycle histamine rate assessment in vaginal content is made. Assessment's result is compared with rate of histamine which is 0.50±0.07 at early estrus stage, 0.34±0.04 at late estrus stage, 0.68±0.08 at metaestrus stage, 0.72±0.12 at early diestrus stage, 0.75±0.13 at late diestrus stage, 0.87±0.11 at proestrus stage. Measurement unit is conventional unit of measuring device. When rate of histamine in vaginal content is below control value the rate of histamine in epithelial cells of endometrium is assessed as reduced, if it exceeds control value same does the histamine rate in endometrial cells.

EFFECT: improvement of evaluation accuracy.

2 tbl, 2 ex

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely to forensic medicine, and can be used for late postmortal detection of prescription of death coming. The optical density of vitreous body is measured. The prescription of death coming is calculated provided a corpse kept at an ambient temperature +10°C to +20°C by formula: PDC=1.975+5.641×OD335+13.924×OD405-16.023×OD475+6.938×OD545, where PDC is the prescription of death coming, (days); OD335 is the optical density of vitreous body at wavelength 335 nm; OD405 is the optical density of vitreous body at wavelength 405 nm; OD475 is the optical density of vitreous body at wavelength 475 nm; OD545 is the optical density of vitreous body at wavelength 545 nm, and at ambient temperature +21°C to +30°C by formula: PDC=1.129+136.486×OD365-134.091×OD375+10.269×OD405, where PDC is the prescription of death coming, (days); OD365 is the optical density of vitreous body at wavelength 365 nm; OD375 is the optical density of vitreous body at wavelength 375 nm; OD405 is the optical density of vitreous body at wavelength 405 nm.

EFFECT: method is accessible, simple and characterised by high accuracy.

2 ex

FIELD: medicine.

SUBSTANCE: tumour region and surrounding intact tissue receives the diagnostic exposure prior to the series low-intensity lasing source exposure that is carried out at average radiation power 1-10 mWt, wavelength 370 nm with recording the tissue-escaping intensity of back-scattered fluorescence at wavelengths 420, 460 and 520 nm. Thereafter, the tumour region and the surrounding intact tissue are exposed to the low-intensity lasing fluorescence at wavelength 532 nm with recording the tissue-escaping intensity of back-scattered fluorescence at wavelengths 560 and 608 nm. Then the immunohistochemical analysis additionally precedes the treatment with recording the Ki-67 antigen expression level of that tumour region exposed to the low-intensity lasing to determine a prediction criterion P1 by the initial Ki-67 antigen expression level; there is evaluated the relation of fluorescence intensities at wavelengths 420/460 nm, 460/520 nm, 560/608 nm derived from the tumour surface Ii:K2=I420/I460, K3=I460/I520, K4=I560/I608 and in the intact region I'i:K'2=I'420/I'520, K'3=I'460/I'520, K'4=I'560/I'608 to calculate the relation of the tumour/intact tissue fluorescence intensity at wavelengths 420/460 nm, 460/520 nm, 560/608 nm: K2/K'2, K3/K'3, K4/K'4; if the Ki-67 antigen expression > 65%, the prediction criteria P1 are considered to be equal to +1, the expression 45-65% shows P1=0, the expression <45% requires P1=-1; the prediction criteria P2, P3, P4 are derived from the results: P2, P3, P4=+1 respectively at K2/K'2<1, K3/K'3<1, K4/K'4<0.5, P2, P3, P4=0 respectively at K2/K'2=1-4, K3/K'3=1-1.4 K4/K'4=0.5-1.5, P2, P3, P4=-1 respectively at K2/K'2>4, K3/K'3>1.4, K4/K'4>1.5, while the radiation sensitivity is determined by total prediction criteria, and P1+P2+P3+P4>0 indicate high radiation sensitivity, P1+P2+P3+P4=0 ensures moderate radiation sensitivity, while in P1+P2+P3+P4<0, low radiation sensitivity is observed.

EFFECT: method simplifies prediction of the results prior to the therapeutic course by tumour sensitivity to ionising radiation, improves its reliability that allows correcting the management program.

1 tbl, 3 ex

FIELD: medicine.

SUBSTANCE: patient's blood serum is tested by infrared spectroscopy. It involves recording peak heights of absorption bands in the range 1170-1025 sm-1, evaluating absorption spectrum, expressing the related values as the regions of "disease patterns" in a coordinate system (x; y) with the coordinates: (x // y): -1.8325/2.2800; -1.3175/2.8650; -1.1468/1.7903; -1.1887/1.3774; -1.0694/0.4032; -1.8500/0.4000; -1.8242/1.6452 - showing clinical effectiveness, and the coordinates: (x // y): 1.1575/2.1000; -1.1274/1.7774; -1.1726/1.3710; -1.0575/0.4200; -0.3775/0.8900 indicating lack of effect.

EFFECT: more accurate and accelerated discovery of estimation of clinical effectiveness.

1 dwg, 2 ex

FIELD: medicine.

SUBSTANCE: blood plasma is analysed for cytochrome oxidase activity; concentrations of 2,3-diphosphoglycerate and lactic acid in erythrocytes are evaluated. Then oxygenation coefficient K is calculated by formula: K = (C1+C2):A, where A is plasma cytochrome oxidase activity, mol/l; C1 - concentration of 2,3-diphosphoglycerate in erythrocytes, mol/l; C2 - concentration of lactic acid in erythrocytes, mol/l. If said oxygenation coefficient is more than 1.0, hypoxia is diagnosed.

EFFECT: technique allows improving reliability of hypoxia diagnostics and reducing acquisition time to 4 hours.

1 ex

FIELD: medicine.

SUBSTANCE: inventions are related to medical diagnostics and may be used to detect concentration of analysed substance, such as glucose, cholesterol, free fatty acids, triglycerides, proteins, ketones, phenylalanine or ferments, in physiological liquid, such as blood, plasm, saliva, urine, interstitial and/or intracellular liquid. Test -element includes the first and second surfaces, which are located at a distance from each other. Surfaces have substantially two identical and mainly congruently matched patterns, which create sections with high and low surface energy on the first and second surfaces, which form system of sample distribution at least with two sensitive sections for measurements. Physiological liquid is held by sections with high surface energy. Device versions are disclosed for performance of analysises, which comprises multiple test-elements, as well as method for making of test-elements, system of analysis for detection of analysed substance concentration in physiological liquid and method for detection of concentration of at least one analysed substance in sample.

EFFECT: provision of simple means and methods for analysis with application of small volume amounts of physiological liquid.

43 cl, 19 dwg

FIELD: medicine.

SUBSTANCE: invention concerns medicine and can be used in dermatology for diagnostics of psoriasis. To implement the diagnostic technique, clinical signs of psoriasis are determined, and as an additional laboratory criterion, spend an chemiluminescence (CL) of low-density and very low-density lipoproteid suspension recovered from the patient's blood serum. It is ensured by measuring sum of light - S, fast flash intensity - I and calculating the coefficient H equal to S to I relation. If H is more than 4.8, psoriasis is diagnosed for the examined patient.

EFFECT: application of the invention allows for objective diagnostics of psoriasis.

2 tbl, 1 ex

FIELD: medicine.

SUBSTANCE: invention concerns medicine and can be used in dermatology for diagnostics of psoriasis. To implement the diagnostic technique, clinical signs of psoriasis are determined, and as an additional laboratory criterion, chemiluminescence (CL) of high-density lipoproteid suspension (HDLS) recovered from the patient's blood serum is evaluated. It is ensured by measuring sum of light (S) and fast flash intensity (I) and calculating the coefficient H equal to S to I relation. If H is more than 5.2, psoriasis is diagnosed for the examined patient.

EFFECT: application of the invention allows for objective diagnostics of psoriasis.

2 tbl, 1 ex

FIELD: medicine.

SUBSTANCE: method consists in estimate of allergic status by infrared spectroscopy of serum sample of venous blood. The blood sample is dried up, powdered to prepare suspension in liquid petrolatum with infrared spectroscopy within 1200-1000 cm-1 to determine peak heights of absorption band at maxima 1170 cm-1, 1165 cm-1, 1160 cm-1, 1150 cm-1 and 1140 cm-1. The coefficients x, y and z are calculated at rotation of axis x by 29°, axis y by 55° and axis z by -1°. The obtained three-dimensional distribution is projected on frontal plane to receive two-dimensional coordinates as diagnostic polygons, and the values (x, y) -0.8431; 1.2793 // -0.1328; 0.7552 // 0.1017; 0.3493 // 0.0397; 0.1414 // -0.9259; 0.8172 allows diagnosing bronchial norm.

EFFECT: higher accuracy and diagnostic objectivity by eliminating subjective interpretation of disease signs.

1 dwg, 5 ex

FIELD: medicine.

SUBSTANCE: early diagnostic technique for cerebral ischemia of various severities in newborns is ensured by determining concentration of average molecular peptides in supernatant fraction of biological fluid of nasopharyngeal aspirate. If the concentration of average molecular peptides is 0.386-0.714 optical density units, mild cerebral ischemia is diagnosed. The concentration 0.715-0,785 optical density unit indicates moderate cerebral ischemia. While at the concentration 0.786-0.853 optical density unit, cerebral ischemia is considered to be heavy.

EFFECT: application of the invention allows detecting cerebral ischemia of various severities at the early stage of disease development in newborns that allows for well-timed treatment-and-prophylactic actions.

4 ex

FIELD: medicine, analytical biochemistry.

SUBSTANCE: invention relates to laboratory methods of investigations. Method involves sampling specimen from patient to be inspected, extraction of serotonin and histamine from a specimen, chromatography of extract and determination of concentration of serotonin and histamine by the fluorescence intensity value. Saliva is used as biological fluid. Saliva by volume 1 ml is extracted with 4 ml of 1 N hydrochloric acid solution, 2 g of anhydrous potassium carbonate and 5 ml of mixture of butanol and chloroform in the ratio 3:2 are added, extract is shaken up and centrifuged. Organic phase (4 ml) is sucked off from extract and passed through chromatography column (diameter is 3 mm, height is 16 mm) filled with ion-exchange resin KB-4 or KB-4P-2 or Bio Rex-70 in H+-form, size of granules is 0.1 ± 0.02 mm. Histamine is eluted with 4 ml of 0.1 N hydrochloric acid at the rate of eluting solution 0.4 ml/min. Histamine concentration is determined by reaction with ortho-phthalic aldehyde dissolved in ethanol. Serotonin concentration is determined by reaction with ninhydrin in organic passed through column. Method provides assaying the saliva concentration of serotonin and histamine with high precision.

EFFECT: improved assay method.

FIELD: medicine.

SUBSTANCE: method involves studying blood samples with venous blood mixed with vital stain like methylene blue. Degree of vital stain absorption by erythrocytes is determined by applying photocolorimetry. The value drop being more than 25%, extracorporal detoxication is to be predicted as ineffective.

EFFECT: simplified method.

6 tbl

FIELD: medicine, infectology.

SUBSTANCE: one should detect the level of terminal stable metabolites of nitrogen oxide (NOx) in whole blood. At its value ranged 39.6-86.0 mcM/l one should evaluate hemorrhagic fever accompanied with renal syndrome (HFRS) of average severity, at NOx value ranged 86.7-141.5 mcM/l - severe form of HFRS and at its values ranged 88.2-128.6 mcM/l at the background of pronounced clinical picture - as complicated disease flow. The method enables to shorten the terms for carrying out the assays.

EFFECT: higher accuracy of evaluation.

3 ex, 1 tbl

FIELD: medicine, biochemistry.

SUBSTANCE: in blood serum one should detect the level of lactoferrin and biliary acids. At their ratio being equal to 5-17 it is necessary to detect chronic hepatitis of high activity.

EFFECT: higher accuracy of detection.

3 ex

FIELD: medicine, dermatology, clinical laboratory diagnostics.

SUBSTANCE: the present method deals with detecting the focus of neutrophilic phagocytic activity lesion in capillary blood. At the values of cells' capacity to phagocytosis in percentage and phagocytic number on the 10th d of therapy being below 20% and 3.3, correspondingly one should evaluate therapeutic efficiency to be low, if it is above 40% and 4.0, correspondingly - as high.

EFFECT: higher accuracy of evaluation.

2 ex, 3 tbl

FIELD: medicine.

SUBSTANCE: method involves studying blood serum, processing obtained data and setting disease diagnosis. The study is carried out by preparing dried blood serum sample as suspension in Vaseline oil and doing the infrared spectroscopy analysis in the bandwidth of 120-1000 cm-1 and determining absorption strip peak heights having maximum at 1180; 1165; 1160; 1150; 1130; 1070; 1025 cm-1 and then calculating the following two ratio groups, the first of which is ratio of peak height with maximum at 1165 cm-1 to 1150 cm-1; 1160 cm-1 to 1130 cm-1; 1070 cm-1; 1025 cm-1. The second group has ratio peak having maximum at 1165 cm-1 to 1160 cm-1; 1180 cm-1 to 1130 cm-1; 1065 cm-1; 1070 cm-1. The obtained three-dimensional distribution of the first group is projected to frontal plane for calculating two-dimensional coordinates and comparing to flat reference diagnostic images of hepatic pathologies and to a normal reference diagnostic image represented as flat polygons which boundaries are given by the following values. The norm is represented by X(-2.3;2.0;4.0;4.0) and Y(1.6;0.8;0.8;1.6), respectively. Oncology is represented by X(1.7;1.7;0.0;0.0) and Y(1.9;1.25; 1.25;1.9). Hepatites are represented by X(1.9;2.2;1.8;1.4 and 1.9;1.8;4.0) and Y(1.9;1.9; 0.5;0.5 and 08;0.5;0.8). Cirrhosis is represented by X(1.9;2.6;1.4) and Y(1.6;0.8;0.4). Diseases are differentiated by interpreting point position within particular area. Three-dimensional distribution of the second group is projected to frontal plane and compared to diagnosis images of pathology and norm. Coordinate values of the second group are as follows: norm - X(1.8;2.9;2.5;1.5), Y(2.7;2.0;1.2;1.6); oncological cases - X(0.27;0.67;0.63), Y(0.27;0.67;0.3); hepatitis - X(1.5;2.5;2.4;1.2), Y(1.6;1.2;0.2;0.9); cirrhosis - X(1.1;0.9;0.9). Final diagnosis of pathology is set when particular data values belong to the corresponding pathology zone in both cases.

EFFECT: high accuracy of diagnosis.

FIELD: medicine.

SUBSTANCE: method involves studying biological material by applying infrared spectroscopy techniques. The obtained data are processed and diagnosis is set. Blood serum is used as the biological material. The study is carried out by preparing dried blood serum sample as suspension in Vaseline oil and doing the infrared spectroscopy analysis in the bandwidth of 120-1000 cm-1 and determining absorption strip peak heights having maximum at 1170; 1165; 1160; 1150; 1140; 1060; 1050; 1040; 1025 and then calculating the following ratio values like peak height with maximum at 1160 cm-1 to 1140 cm-1; 1165 cm-1 to 1150 cm-1; 1040 cm-1 to 1025 cm-1. The obtained distribution of this group is projected to frontal plane for calculating two-dimensional coordinates and comparing to flat reference diagnostic images of prostate pathologies and to a normal reference diagnostic image represented as flat polygons which boundaries are given by the following values. The norm is represented by X(-1.15;-0.9;0.45;0.0;-0.65) and Y(0.99;4.2;0.9;0.46), respectively. Pathology by X(-1.15;-1.15;0.35;0.0;0.65) and Y(0.99;-0.03; 0.48;0.09;0.46). The norm and pathology are differentiated. Additional mathematical processing is carried out on infrared spectra of blood serum samples of patients belonging to pathology image according to parameter values. First of all, three-dimensional distribution is calculated as peak having maximum at 1160 cm-1 to one having maximum at 1150 cm-1; 1170; 1160 cm-1; 1160 cm-1 to 1025 cm-1. It is projected then to frontal plane and compared to diagnosis images of prostate adenoma and images of prostate carcinoma. The second group relationships the following values are used: oncological cases - X(0.28;0.77;1.24;0.96), Y(0.75;0.46;-0.13;-0.02); adenoma - X(0.28;1.24;2.21;1.24;0.77), Y(0.75;1.24;-0.12;-0.13;0.46). Differential diagnosis of pathologies is set by interpreting point position within particular pathology image.

EFFECT: high accuracy of differential diagnosis.

FIELD: medicine.

SUBSTANCE: method involves determining mean cytochemical coefficient of lipid accumulation in peripheral blood leukocytes in conditional units before beginning therapy application (MCC1) and in 2-3 or 5-6, or 10-12, or 20-24 months of therapy application (MCC2). Therapy effectiveness coefficient is calculated in conditional units from formula K= MCC2/MCC1. The value being equal to or greater than 1, leprosy therapy is predicted to be effective.

EFFECT: simplified prognosis method.

1 dwg, 1 tbl

FIELD: medicine.

SUBSTANCE: method involves determining infrared radiation absorption coefficient in blood plasma in bandwidth of 1543-1396 cm-1. The infrared radiation absorption coefficient is determined in %. The value being equal to 29.7±1.1%, catarrhal cholecystitis is diagnosed. The value being 26.4±1.4%, phlegmonous cholecystitis is diagnosed. The value being 21.2±1.8%, gangrenous cholecystitis is diagnosed. The value being equal to 18.6±0.5%, gangrenous perforated cholecystitis case is diagnosed. The value in norm is equal to 32.4±0.8%.

EFFECT: high accuracy and specificity of diagnosis.

FIELD: medicine.

SUBSTANCE: method involves pouring venous blood treated with heparin into five conic test-tubes in the amount of 0.1 ml. The first three of them contain 0.1 ml of non-colored latex suspension with particle size of 1.5 mcm, the fourth one contains 0.1 ml of medium 199 and 0.1 ml of 0.1% aqueous solution of tetrazole nitro blue, the fifth one contains .1 ml of latex suspension and 0.1 ml of 0.1% aqueous solution of tetrazole nitro blue. The first test-tube is incubated in thermostat for 5 min at37°C, the second one for 30 min, the third one for 1 h, the fourth and the fifth one for 40 min. Smears are prepared from 0.2 ml of incubation mixture on glasses and dried at 37°C, fixed in burner flame, stained with 0.1% aqueous solution of tetrazole nitro blue, repeatedly dried and studied with microscope under immersion with magnification of 90x10. Test results are evaluated from absorption activity in phagocytosis reactions in determining the number of phagocytes, phagocytic number, phagocytic integral index and phagocytosis rate values. Tetrazole nitro blue test response is determined by counting formazan-positive cell number, calculating cytochemical activity index and tetrazole nitro blue test stimulation index.

EFFECT: accelerated test; high accuracy and low cost of examination.

1 dwg, 3 tbl

Up!