Method of predicting risk of glomerular filtration rate reduction after aortocoronary bypass surgery on working heart
SUBSTANCE: invention relates to medicine, namely to a method of predicting a probability of reduction of the glomerular filtration rate (GFR) after 3 months of observation after aortocoronary bypass surgery without artificial blood circulation (ACBS without ABC). The essence of the method consists in the fact that the concentration of a kidney injury molecule of type 1 (KIM-1) is determined in blood serum, the ratio of the biomarker KIM-1 concentrations in two time points after 48 hours and 7 days after the operation is calculated and if its value is higher than 1.5, a conclusion about the probability of (GFR) reduction in the remote period after ACBS without ABC is made.
EFFECT: application of the claimed method makes it possible to predict the probability of the glomerular filtration rate (GFR) reduction after 3 months of observation after aortocoronary bypass surgery without artificial blood circulation in an efficient and accurate way.
1 tbl, 1 dwg, 1 ex
The invention relates to medicine, namely to surgery and Nephrology, and can be used in the departments of cardiac surgery, therapy, as well as in outpatient clinics (outpatient medical care).
Coronary artery bypass grafting (CABG) has established itself as an effective method of treatment of coronary heart disease (CHD). Procedure on a beating heart without cardiopulmonary bypass, without IR) is considered preferable because of the smaller risk of developing acute kidney injury (AKI), systemic inflammatory response and other adverse effects of IR [1-3]. The search for early predictors of AKI after cardiac surgery and to assess their informativeness in comparison with the classical marker of renal function - serum creatinine (TFR) [4-6]. Based on the values of the TFR one of the estimated means estimated glomerular filtration rate (GFR). Alternatively, the TFR and GFR are studied biomarkers: cystatin C, interleukin-18, kidney injury molecule type 1 (kidney injury molecule-1 KIM-1) and others [7-9]. It should be noted that in most studies the observation period is limited to 48 to 72 hours of the postoperative period [11, 12], i.e. as the time point chosen point of diagnosis of AKI. Information about the dynamics functions almost�to the long period of observation (months - years) in patients undergoing CABG without IR, almost never occur, despite the fact that the presence of chronic kidney disease associated with increased cardiovascular risk and reduces the lifespan. KIM-1 is a surface protein of the immunoglobulin superfamily, is localized predominantly in the apical membrane of proximal epithelial cells is recognized as one of the most sensitive markers of AKI [5, 12]. After damage to renal tubular KIM-1 begins to be excreted with the urine. Investigated the concentration of KIM-1 in urine and serum . High levels suggest an adverse outcome of AKI . In addition, KM-1 impact on preserving the viability of epithelial cells and turns them into phagocytes, able to absorb dead cells, which greatly speeds up the process of recovery of renal function . Thus, KIM-1 is not only a diagnostic marker, but also increases the intensity of the processes of recovery of the structure and function of the renal tissue. Data on the dynamics of the levels of KIM-1 and their prognostic value in long-term monitoring of renal function after specific techniques CABG without IR is very limited.
Objective: to evaluate the possibility of using early postoperative level of KIM-1 as biomarker for prediction of GFR decline che�ez 3 months of follow-up after CABG without IR.
Based on the research model developed a method of predicting deterioration in renal function (decrease GFR) in the remote period after CABG without PC. The analogue of the developed model is the work of , which the authors suggest the use of early postoperative levels of KIM-1 and hepatic protein associated with fatty acids (liver fatty acid-binding protein, L-fabp gene) to predict AKI after cardiac surgery (not only after isolated CABG techniques without IR). The analogue model is based on the levels of 2 biomarkers (KIM-1 and L-fabp gene) measured in the first 6-12 hours after surgery, that is more time-consuming. Dunn model involves the measurement of a single biomarker KIM-1 in two points. The area under the ROC curve of the model-equivalent to 0.78, while for the author of the model area is 0,818, which means that the latter has a greater predictive power. Besides the authors of the analog propose a method for predicting AKI (develops within 48-72 hours after surgery), not considering the long-term dynamics of renal function. The proposed prediction method is aimed at assessing the risk of deterioration in renal function (decrease GFR) in the long period of follow-up after CABG without IR.
Description of the method: to determine the risk of an impaired GFR at 3 months after CABG without IR 48 hours � 7 days after surgery to determine the serum level of KIM-1 and the ratio of the levels of KIM-1 in the above points and if you value this relationship more than 1.5 make a conclusion about the risk of decline in GFR after 3 months, that is about the risk of distant worsening of renal function. Prediction probability (sensitivity 82%, specificity of the method was 81%. The predictability of a positive test result 58%. The predictability of a negative test result 80%. Likelihood ratio for a positive result is 4.3.
Positive effect: prediction of GFR decline allows in the early postoperative period to allocate the risk group for targeted monitoring of threatened patients after CABG without IR, regular assessment of renal function is available for outpatient methods (levels of TFR and GFR) and when indicated in a timely manner to refer patients for a consultation with a nephrologist. When using the proposed method of risk assessment in the specified group may be prescribe drugs with nephroprotective properties and to avoid unreasonable prescription of nephrotoxic funds. For example, after cardiac surgery are often prescribed an antibacterial agent that can reduce GFR, especially in patients with compromised renal function (aminoglycosides, and some cephalosporins). In this case, it is possible to avoid unreasonable prescription of medicines or the use of less nephrotoxic funds. When disturbed function almost�to undesirable assignment potassium-sparing diuretics (spironolactone). When the need for diuretics is at risk, you should use a loop or thiazide diuretics. An indication of the risk of an impaired GFR at discharge from the Department of cardiac surgery may be useful for alertness outpatient clinics in terms of a possible deterioration in renal function remote period after CABG surgery without IR. In addition, information about possible reductions in GFR after CABG without ROS will be useful for secondary prevention of coronary heart disease, since the development of CKD and the growing severity indicates an increase in cardiovascular risk.
Material and methods. The study included 30 patients with ischemic heart disease, including 23 men (76.7 percent) and 7 women (23.3 percent) who had undergone isolated CABG without IR, mean age 57.9±4.7 years.
After 48 hours and 7 days after CABG without IR were measured the levels of TFR and GFR according to standard procedures. At the same time points by ELISA using commercial reagents were determined serum levels of KIM-1. On the basis of the TFR 48 hours after CABG without IR AKI was diagnosed in 3 cases (10%). The decrease in GFR at 3 months after surgery occurred in 11 patients (36,7%), i.e. every 3-4th. The initial level of KIM-1 was 30.8 (11,0-65,6) ng/ml.
It was calculated the ratio of the concentrations of KIM-1 after 48 hours and 7 days after CABG without IR: KIM-148 hours/KIM-17 �it - Method of constructing ROC curves is determined that the value of this ratio of 1.5 and above with a sensitivity of 82% and specificity of 81% to predict the decrease in GFR at 3 months after CABG without IR (Fig.1). The decrease in GFR - 17.3 ml/min/1.73 m2(17%) (95% CI 8-26), t (3, 27)=4,22, p=0.001.
The data obtained confirms the method of logistic regression: the attitude of KIM-148 hours/KIM-17 dayswas a predictor of outcome (GFR decrease/increase) after 3 months of observation after surgery (table 1).
|Predictors of GFR change (decrease/increase) 3 months after CABG without IR (n=27)|
|B(SE)||95% CI for the odds ratio (OR)||P|
|KIM-148 hours/KIM-17 days||-0,940 (0,428)||0,169||0,390||0,903||0,028|
|Note. To reduce GFR: R2=0,285 (Cox and Snell); R2=0,385 (Nagelkerke). Model χ2=9,062, p=0.003|
Clinical example of the use of the method
Patient M., 54 years old, suffering from coronary heart disease, surgery performed isolated CABG without IR in the cardiac surgery Department of pathology Department of the Arkhangelsk region "the First city clinical hospital named. E. E. Volosevich" in 2012.
Baseline renal function was normal with the levels of TFR and GFR 82,0 µmol/l and 90,4 ml/min/1.73 m2respectively. The level of KIM-1 was determined in the serum during the day before surgery (to 57.2 ng/ml) and 48 hours (101,9 ng/ml) and 7 days (24,0 ng/ml) after CABG without IR. The attitude of KIM-148 hours/KIM-17 gnawpatient M. was 4.25, we have exceeded the proposed diagnostic value (1,5), on what basis it was concluded that the risk of an impaired GFR at 3 months after surgery.
This forecast was confirmed. After 3 months of observation in the clinic has seen a worsening of renal function: GFR decline by 16.7 ml/min/1.73 m2(18.5%), - actual GFR amounted to 73.7 ml/min/1.73 m2.
This clinical case confirms the positive effect when using the proposed method.
Relative�Linux serum concentrations of KIM-1, defined in 48 hours and 7 days after surgical treatment, can be used as a marker for determining the risk of an impaired GFR in the remote period after CABG without IR.
The list of sources
1. Nigwekar SU, Kandula P, Hix JK et al. Off-pump coronary artery bypass surgery and acute kidney injury: a meta-analysis of randomized and observational studies. American Journal of Kidney Diseases 2009; 54(3): 413-423.
2. Massoudy P, Wagner S, Thielmann M, et al. Coronary artery bypass surgery and acute kidney injury - impact of the off-pump technique. Nephrology, Dialysis and Transplantation 2008; 23 (9): 2853-2860.
3. Nair S, Iqbal K, Phadke M et al. Effect ofcardiopulmonary bypass on tissue injury markers and endothelial activation during coronary artery bypass graft surgery. Journal Postgraduate Medicine 2012; 58 (1): 8-13.
4. Eren Z, Ozveren O, Buyukoner E et al. A Single-centre study of acute cardiorenal syndrome: incidence, risk factors and consequences. Cardiorenal Medicine 2012; 2: 168-176.
5. Huo W, Zhang K, Nie Z et al. Kidney injury molecule-1 (KIM-1): a novel kidney-specific injury molecule playing potential double-edged functions in kidney injury. Transplantation Reviews 2010; 24: 143-146.
6. Sprenkle P, Russo P. Molecular markers for ischemia, do we have something better then creatinine and glomerular filtration rate? Arch Esp Urol 2013; 66(1): 99-114.
7. Che M, Xie B, Xue S et al. Clinical usefulness of novel biomarkers for the detection of acute kidney injury following elective cardiac surgery. Nephron Clinical Practice 2010; 115: 66-72.
8. Tsigou E, Psallida V, Demponeras C et al. Role of new biomarkers: functional and structural damage. Crit Care Res Pract 2013 February 5: [Epub ahead of print].
9. Parikh CR, Thiessen-Philbrook H, Garg AX et al. Performance of kidney injury molecule-1 and liver fatty acid-binding protein and combined biomarkers of AKI after cardiac surgery. Clin J Am Soc Nephrol 2013 Apr 18. [Epub ahead of print].
10. Sidebotham D. Novel biomarkers for cardiac surgery-associated acute kidney injury: a skeptical assessment of their role. J Extra Corpor Technol 2012; 44 (4): 235-240.11. Han WK, Wagener G, Zhu Y et al. Urinary biomarkers in the early detection of acute kidney injury after cardiac surgery. Clin J Am Soc Nephrol 2009; 4 (5): 873-882.
12. Huo W, Zhang K, Nie Z et al. Kidney injury molecule-1 (KIM-1): a novel kidney-specific injury molecule playing potential double-edged functions in kidney injury. Transplantation Reviews 2010; 24: 143-146.
13. Han WK, Bailly V, Abichandani R, et al. Kidney Injury Molecule-1 (KIM-1): a novel biomarker for human renal proximal tubule injury. Kidney Int 2002; 62(1): 237-44.
14. Liangos O, Perianayagam MC, Vaidya VS, et al. Urinary N-acetyl-beta-(D)-glucosaminidase activity and kidney injury molecule-1 level are associated with adverse outcomes in acute renal failure. J Am Soc Nephrol 2007; 18 (3): 904-912.
15. Ichimura T, Asseldonk EJ, Humphreys BD et al. Kidney injury molecule-1 is a phosphatidylserine receptor that confers a phagocytic phenotype on epithelial cells. J Clin Invest 2008; 118 (5): 1657-1668.
A method of predicting the likelihood of a reduction in glomerular filtration rate (GFR) after 3 months of follow-up after coronary artery bypass grafting without extracorporeal circulation (CABG without IR), including the determination of the concentration of biomarker of kidney injury molecule type 1 (KIM-1), characterized in that in the serum to determine the concentration of KIM-1, calculate the ratio of concentrations of the biomarker KIM-1 in two time points after 48 h and 7 days after surgery and when the value of more than 1.5 predict the likelihood of decreased GFR in the remote period after CABG without IR.
SUBSTANCE: early diagnostic technique for pulmonary thromboembolism consists in the fact that if the patient shows any clinical signs indicative of the probability of pulmonary thromboembolism, blood serum fibrin D-dimer is supposed to be measured; if the fibrin D-dimer is less than 0.5 mg/l, a suspected case of pulmonary thromboembolism is cleared of, whereat blood serum fibrin D-dimer of 0.5 mg/l and more requires measuring blood serum interleukin-8 additionally by enzyme immunoassay, and if the measured concentration is 21.3 pg/ml and more, pulmonary thromboembolism is diagnosed.
EFFECT: more accurate and informative diagnostics.
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention relates to the field of biotechnology, namely to obtaining oligopeptide compounds, containing a motive, interacting with a proliferating cell nuclear antigen (PCNA) and can be used in medicine. The oligopeptide compound consists of 14-70 amino acids and contains. a PCNA-interacting motive, representing [K/R]-[F/Y/W]-[L/I/V/A]-[L/I/V/A]-[K/R], at least one signal sequence of nuclear localisation and at least one signal sequence of penetration into a cell, with the PCNA-interacting motive being located towards an N-end relative to the signal sequence.
EFFECT: invention makes it possible to carry out the efficient treatment of hyperproliferative disorders by the application of the oligopeptide compound in cyctostatic therapy or in radiotherapy as a sensitising substance.
34 cl, 6 dwg, 4 tbl, 8 ex
SUBSTANCE: on paramagnetic particles bearing the immobilised bacterial protein G of the family Streptococcus, with the blocked solution Denhardt-DNA the protein BoNT/A is adsorbed using specific highly affinity polyclonal antibodies. The formation of the protein complex BoNT/A is detected with a biotin-conjugated antibody by a noncovalent conjugate of DNA fragments with neutravidin. PCR amplification of the DNA matrix is carried out with the fluorescence detection of the signal in real time. Registration of the presence of BoNT/A in the samples under study is carried out by the change in the level of fluorescence compared to the control ones.
EFFECT: effective method of determining the presence of the compound.
5 dwg, 1 tbl, 5 ex
SUBSTANCE: invention relates to the field of medicine, namely to a method of predicting the development of severe sepsis. The essence of the method consists in the following: on the first day after acute poisoning with substances of narcotic action the content of procalcitonin, interleukin-6 and interleukin-8 is determined in blood serum. If the value of procalcitonin is from 0.5 ng/ml to 5 ng/ml, interleukin-6 from 20 pg/ml to 150 pg/ml, interleukin-8 from 50 pg/ml to 300 pg/ml, the development of severe sepsis is predicted.
EFFECT: application of the claimed method ensures the high reliability of prediction of severe sepsis development.
1 tbl, 3 ex
SUBSTANCE: invention relates to the field of molecular genetics, genosystematics and pharmacognosy and is intended for the identification of the species affinity of common heather (Calluna vulgaris (L.) Hull.). Claimed is a set of synthetic oligonucleotides for PCR with the fragment ITS2 of nuclear DNA, including forward and reverse primers and a degradable probe.
EFFECT: set possesses high sensitivity and specificity and makes it possible to carry out the identification of a medicinal plant in a fast and reliable way.
1 dwg, 1 ex
SUBSTANCE: invention aims at detecting benign and malignant new growths in human thyroid. Involved thyroid and reference adjacent intact tissues are sampled; micro-RNA is recovered from the samples; that is followed by conducting a reverse transcription reaction, measuring an expression level of microRNA-21, -221, -222, -155, -205 by real-time RNA followed by a comparative analysis of the microRNA expression according to the norm and thyroid tumour involvement, and stating the presence and type of the new growth. If the above microRNA expression varies by no more than 4 times to the higher and lower figures of expression in relation to the reference, the benign new growth is stated. The malignant new growth is shown by the measured microRNA expression by more than 4 times.
EFFECT: effective detection of the benign and malignant thyroid new growths that promotes improving the further therapeutic approach.
5 dwg, 1 tbl, 4 ex
SUBSTANCE: present group of inventions refers to medicine, namely to cardiology, and concerns diagnosing various forms of the left ventricular hypertrophy. That is ensured by determining an amount of a necrosis marker - cardiac troponin T, cardiac function marker - NT-proBNP and one of an inflammation marker - GDF-15. Comparing their amounts to the reference values enables diagnosing a physiological or pathological left ventricular hypertrophy. Besides, the presence of the pathological hypertrophy requires determining the relation between the cardiac function marker and inflammatory marker, as well as between the necrosis marker and inflammatory marker, and comparing these relations to the reference enables stating if the individual suffers from hypertrophic nonobstructive cardiomyopathy, or hypertrophic obstructive cardiomyopathy, or hypertrophy accompanying pressure overload.
EFFECT: improving the diagnostic procedure.
7 cl, 3 tbl, 3 ex
SUBSTANCE: method of detection of intracellular proteins using fluorescein-5-isothiocyanate (FITC) in various types of mammalian cells is provided, which are characterised by different levels of the synthetic process, using the method of confocal laser microscopy. The mammalian cells are fixed by paraformaldehyde (PFA) preventing the extraction of proteins in subsequent stages of staining, permeabilised with detergent, then stained for 2 h with FITC in a concentration of 1 mcg/ml. The cells are enclosed in Mowiol 4-88 adding DABCO (1,4-diazabicyclo[2.2.2] octane). The resulting preparations are used to analyse the localisation and the content of the protein component of the cells by confocal microscopy.
EFFECT: invention enables to obtain information and to investigate the location of proteins in the cells and to carry out a comparative analysis of protein content in cells with different levels of metabolism.
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SUBSTANCE: if observing clinical risk factors of thromboembolia of the pulmonary artery, venous blood 3 ml is taken from the patient to analyse polymorphisms -174 G>C of IL-6 gene and -308 G>A of TNFalpha gene by polymerase chain reaction. The genetic typing results are determined by assigning certain points: if observing GG genotype of IL-6 gene, 3 points are assigned, GC - 1 point, CC - 0 points; the GG genotype of TNFalpha gene, 3 points are assigned, GA - 1 point, AA - 0 points, the total score of certain patient is derived. If the total score is 4 and more, the presence of individual predisposition to thromboembolia of the pulmonary artery is stated.
EFFECT: invention provides the effective assessment of genetic predisposition to thromboembolia of the pulmonary artery.
2 tbl, 2 ex
SUBSTANCE: observing no development of ventriculomegalia on the 5th day is combined with the postnatal seromucoid concentration of 0.136-0.166 absorbance units measured in supernatant of biological nasopharyngeal aspirate, whereas ventriculomegalia developing on the 5th day is shown by the postnatal seromucoid concentration of seromucoid concentration of 0.167-0.196 absorbance units measured in supernatant of biological nasopharyngeal aspirate.
EFFECT: early diagnosis of ventriculomegalia accompanying moderate cerebral ischemia caused by the intrauterine cytomegaloviral-herpetic infection in newborns.
2 tbl, 3 ex
SUBSTANCE: method includes determination of a titre of antibodies to cytomegalovirus, content of 2,3 DPG (2,3-diphosphoglycerate), and oxyhaemoglobin in erythrocytes. If the titre of the antibodies to cytomegalovirus increases to 1:1600, DPG increases to 6.7±0.3 mcmole/ml, content of HbO2 is 95.0±1.7%, with the reduction of the specific optical density of haemoglobin to 0.70±0.01, it is concluded that a threat of anaemia development is formed.
EFFECT: method makes it possible to study the character of haemoglobin oxygenation impairment by means of determination of the specific optical density.
SUBSTANCE: neurovisualisation examination of brain is carried out, Cirs comorbidity index and Kaplan-Feinstein comorbidity index are determined, cochleovestibular syndrome, eye-moving impairments, type of diabetes mellitus are identified. Value of discriminate function (D) is calculated. If D value is higher than zero, diagnosed are consequences of ischemic brain stroke (IBS) with hyperhomocysteinemia (HH), if D is lower than zero, consequences of IBS without HH are diagnosed.
EFFECT: method makes it possible to increase reliability of diagnostics of IBS consequences, which is achieved due to complex analysis of said parameters.
FIELD: veterinary science.
SUBSTANCE: method includes detection of leukocytes, protein, haemoglobin in faeces, and PH-reaction of faeces. A sample of faeces is dissolved in distilled water and applied drop by drop onto appropriate test fields of a test strip designed for urine testing. By variation of colour within 1 minute the reaction is assumed as positive. If pH is less than 7.0 or more than 7.5, and faeces contain soluble protein, haemoglobin and leukocytes at the same time, availability of the inflammatory process in the intestines is confirmed.
EFFECT: invention makes it possible to quickly and accurately diagnose inflammatory process in intestines.
2 cl, 2 ex
SUBSTANCE: invention refers to medicine and aims at diagnosing vaginitis in females of a reproductive age. Expression levels of mRNA of TNF, IL18, GATA3 and TLR4 genes are measured in vaginal smears. Expression levels of mRNA of TLR4/GATA3 and TNF/IL18 genes are related, and the derived threshold cycles are used to predict a probability of vaginitis by formula. If P≤57% for nonpregnant females or P≤68.5% for pregnant females, the absence of vaginitis is stated. If P≥57% for nonpregnant females or P≥68.5% for pregnant females, vaginitis is diagnosed.
EFFECT: invention provides the effective diagnosis of vaginitis in the females of a reproductive age.
3 tbl, 5 ex
SUBSTANCE: group of inventions refers to medicine, namely to a method of analysing ex-vivo if a patient suffering a cancer is expected to respond therapeutically to a method of treating. To this purpose, a biological sample specified in a group consisting of total blood, plasma or serum samples is taken from the patient. The levels IL10 and IFNγ are measured in the above biological sample; the levels are determined with using the respective IL10 and INFγ specific antibodies. The IL10/IFNγ ratio is derived and compared to a threshold. If the IL10/IFNγ ratio of less than 4 shows that the patient is expected to develop the preventive or therapeutic response to the immunogenic composition. The group of inventions also refers to using IL10 and INFγ as biomarkers and a kit for analysing the above method.
EFFECT: using the given method enables predicting the sensitivity of the patient suffering cancer to the therapeutic treatment, as well as obtaining an algorithm for treatment modification for improving the patient's response to the treatment.
12 cl, 2 dwg, 1 ex
SUBSTANCE: invention deals with early diagnostics of infectious complications in patients with acute lymphoblastic leukaemia (ALL), obtaining chemotherapy (CT). Claimed method consists in the following: concentration of fibrinogen, time of XIIa-dependent euglobulin lysis (XIIa-DEL), level of soluble fibrin-monomer complexes (SFMC), activity of protein C (prC) are analysed in ALL patients during CT 2-3 times per week. If at least 2 of 4 criteria are detected, namely: ≥41% increase of fibrinogen level, ≥76% increase of SFMC, ≥11% prolongation of XIIa-DEL time and ≥12% reduction of prC activity in comparison with previous analysis, development of infectious complications is predicted.
EFFECT: method provides early prediction of infection development in patients with acute lymphoblastic leukaemia during chemotherapy.
7 tbl, 4 ex
SUBSTANCE: level of anti-flu antibodies in blood serum from umbilical vein (A), concentration of middle molecular peptides in blood plasma from umbilical vein (units of optical density) (B), total number of T-lymphocytes (CD3+) in points: 1 point - CD3+ higher than 48%; 2 points - CD3+ 47%-41%; 3 points - CD3+ 40% and lower in venous umbilical blood are determined in term new-born babies at birth. After that, prediction of frequent development of acute respiratory viral infections is realised by means of discriminant equation: D=+0.008×A-111.694×B-1.537×C, where D is discriminant function with boundary value, equal - 34.16. If D is equal or is larger than boundary value, absence of development of frequent respiratory viral infections during the first year of life in term new-born babies with intrauterine flu A(H3N2) is predicted. If D is lower than boundary value, frequent development of acute respiratory viral infections during the first year of life in term new-born babies is predicted.
EFFECT: increased probability of correct prediction of frequent development of acute respiratory viral infections during the first year of life in term new-born babies.
SUBSTANCE: biological object, which contains mixture of hydroxybenzene and its monomethyl-substituted are crashed, processed twice with ethylacetate for 30 minutes with ethylacetate, ethylacetate extracts are combined, treated with ethanol solution of calcium hydroxide, solvent from combined ethylacetate extract is evaporated at 16-20°C, residue is repeatedly treated with acetone, containing hydrochloric acid in excess with respect to potassium hydroxide, present of residue, acidified acetone extracts are combined, treated with water solution of sodium hydroxide to neutralise residues of hydrochloric acid in acetone and create excess of alkaline medium, acetone is evaporated from combined extract, water-alkali residue is diluted with water, formed solution is acidified to pH 2-3, saturated with sodium sulphate, extracted with diethyl ether, extract is dehydrated, evaporated, chromatographed in column with silica gel with application of mobile phase hexane-diethyl ether with ratio 6:4 in volume, eluate fractions, containing hydroxybenzene and its monomethyl substituted, are combined, extracted with buffer solution with pH 12-13, water-alkali extract is acidified with 24% solution of hydrochloric acid to pH 2-3, saturated with sodium sulphate, extracted with dichloromethane, dichloromethane extract is dehydrated, analysed substances, contained in dichloromethane extract, are transferred into corresponding trimethylsilyl derivatives, for which purpose dichloromethane extract is treated for 20 minutes with N-methyl-N-trimethylsilyltrifluoracetamide under conditions of heating at temperature 60°C, qualitative and quantitative determination by physical-chemical method, such as chromate-mass-spectrometry, is performed in capillary column, 25 m long with internal diameter 0.2 mm with immobile phase (5%-phenyl)-methylpolysiloxane, with application of helium carrier gas, supplied at rate 0.6 ml/min, and mass-selective detector, working in electron impact mode, initial temperature of column thermostat constitutes 70°C, said temperature is maintained for 3 minutes, and then temperature is programmed from 70°C to 290°C at rate 20°C, final temperature of column is maintained for 10 minutes, injector temperature constitutes 250°C, quadrupole temperature - 150°C, temperature of ion source - 230°C, temperature of detector interface - 300°C, intensity of signal, conditioned by charged particles, formed in bombardment of analysed substance, leaving capillary column and getting into source of ions, with ionising beam of electrons with energy 70 eV, is registered, mass-spectrum is registered by full ion flow, qualitative determination of analysed substance is realised by time of exposure, set and intensity of signals of characteristic charged particles in mass-spectrum of its trimethylsilyl derivative, with quantity of determined compound being calculated by area of chromatographic peak of its trimethylsilyl derivative.
EFFECT: increasing sensitivity and reduction of determination duration.
6 tbl, 1 dwg, 5 ex
SUBSTANCE: invention relates to medicine, namely to method of predicting development of terminal stages in patients with chronic myeloleukaemia. Essence of method consists in the fact that activity of two degydrogenases - glycerol-3-phosphatedehydrogenase (G3PDH) and lactatedehydrogenases (LDG) is determined in lymphocytes of peripheral blood of chronic leukaemia patients in open stage. In case of combination of G3PDH in the range from 0 to 0.02 mcE and activity of LDG in the range from 0.04 to 5.02 mcE development of terminal stage is predicted.
EFFECT: application of claimed invention makes it possible to diagnose progress of disease, correct plan and tactics of treating given category of patients and improve results of their rehabilitation in due time.
SUBSTANCE: invention refers to microbiology, namely to a method for the microbiological examination of a native smear from a tongue root mucosa. The substance of the method consists in the fact that a wooden spatula is used to sample biomaterial 0.5 ml by deep scraping from the mucous membrane at the boundary of the posterior one-third of the tongue dorsum and root; two smears are prepared on two slides, 1.5-2.0 cm2 in area; the smears are dried out for 20 minutes and fixed by flame heat of an alcohol lamp continuously for 5-6 seconds, Gram-stained by a complicated differential method, microscoped with immersion with the use of a ×100 objective lens; at least 10 visual fields are inspected on each slide taking into account quantitative and qualitative characteristics of microorganisms, cells and nuclei of the epithelium.
EFFECT: invention enables the more precise examination of the native smear of the tongue.
4 dwg, 1 tbl, 2 ex
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.