Method for preclinical study of cardiotropic antiarrhythmic drug
SUBSTANCE: invention represents a method for preclinical study of cardiotropic antiarrhythmic drugs, involving determining the bioelectric parameters in isolated multicellular perfused preparations and measuring an action potential duration, differing by the fact that the isolated multicellular perfused preparations are presented by rat's pulmonary vein myocardium; the parameters are measured in three operation modes of the multicellular preparations; a resting potential is additionally measured; varying APD 90%, related APD 50%/APD 90%, a spontaneous shear velocity of the resting potential, the most positive membrane potential in the resting preparation, a spontaneous activity train repetition rate, spontaneous action potential train repetition and variability frequency, post-depolarisation number and intensity, as well as a shear membrane potential corresponding to the beginning of train activity are used to evaluate the signs of antiarrhythmic and arrhythmogenic action.
EFFECT: more reliable prediction of the antiarrhythmic action of the potential pharmacological agents and reduction of experimental phase time.
The present invention relates to experimental biology, in particular for experimental pharmacology and cardiology, concerns preclinical studies cardiotropic antiarrhythmic drugs and other cardiotropic funds, security definitions, i.e., the absence of arrhythmogenic activity of drugs.
At the present time to treat rhythm disorders of various types are widely used antiarrhythmic drugs. Continuously being developed to a considerable number of compounds, claiming new antiaritmikov. The effects of most anti-arrhythmic drugs is multifactorial, implemented at the molecular, cellular and tissue levels: it is due to the influence of molecular membrane and intracellular complexes, the bioelectric activity of individual cardiomyocytes, the electrical properties of cardiac tissue. Antiarrhythmic agent have an impact on all parts of the heart, including pacemaker, work infarction, vascular system, but unequally.
Antiarrhythmic agent, as well as other cardiotropic means cause side effects that may have undesirable properties, for example, under certain conditions, can lead to arrhythmias, i.e., to have arrhythmogenic effect.
In connection with slozhnosti.imenno steps and a plurality of target detection and predicting antiarrhythmic actions as well as side, arrhythmogenic properties of antiarrhythmic (and cardiotropic drugs is an extremely difficult task.
There are factors and scope, critical for the implementation of various types of anti-arrhythmic activity. The most common forms of arrhythmia is atrial flutter and flicker (atrial fibrillation) atrial. According to the most common at the present time the concept of ectopic activity in the field of myocardial plates ("arms") of the pulmonary veins, which interacts with the electrical activity in the Atria, is the cause of atrial fibrillation (AF). Myocardial tissue in the pulmonary veins differs from atrial, has a number of morphological features and characteristics of bioelectric activity. Myocardial sleeves of the pulmonary veins in the experimental conditions, and in vivo are foci triggering activity, abnormal automatie, circulation of excitation, i.e., the factors leading to the different types of arrhythmias, in particular atrial fibrillation.
Changes of bioelectric activity in the myocardium pulmonary veins under the action of pharmacological agents is critical, the Central element of the mechanism of initiation of atrial fibrillation, and prevention.
There are several in vivo and in vitro the ways of identifying, predicting antiarrhythmic activity. This model chloralkali arrhythmias, model gloriamarie arrhythmias, adrenaline, continua model arrhythmias. The essence of these methods is the introduction of high doses of calcium, barium, aconitine, oubain or strofantina anesthetized or awake animals.
(Gabriel R. U. Guidance on experimental (preclinical) study of new pharmacological substances, M.: Medicine, 2005.)
Infusion of calcium chloride to simulate ventricular and supraventricular arrhythmias proposed in the 1950's. Model chloralkali arrhythmias based on a generalized activation of the sympathetic division of the autonomic nervous system and subsequent release of norepinephrine from sympathetic terminala that, in turn, leads to the development of arrhythmia and abnormal automatie. Similar in nature is "adrenaline model arrhythmia", differing from the previous approach only the immediate injection of adrenaline.
(Role of the sympathetic-adrenal system in the mechanisms of genesis of cardiac fibrillation induced by calcium chloride. Kardiologiia. 1974 Jun; 14 (6): 78-82.)
"Hloridnaja model of arrhythmia based on the ability of barium disrupt the functioning of mitochondria, the conductivity of several groups of ion channels.
(Delfino G, Amerini S, Mugelli A. Barium cardiotoxicity: Relationship between ultrastructural damage and mechanica effects. Toxicol In Vitro. 1988; 2 (1): 49-55.)
"Continua model" and "Stroganina model" is based on the systemic administration of specific toxins: aconitine, suppressing the inactivation of sodium potentialcustomers channels of cardiomyocytes, oubain, suppressing the activity of Na/K-ATPase cardiomyocytes, respectively.
(Chan TY. Aconite poisoning. Clin Toxicol (Phila). 2009 Apr; 47 (4): 279-85.)
All of the above approaches practically have nothing to do with the physiological mechanisms leading to the occurrence of arrhythmias in vivo, and induced rhythm disturbances characteristic of the "agonistic" or acute intoxication. Using "aconitine" and "strofantino" models is justified only in the case of the development of products to treat rare genetic conditions. When using the above approaches it is impossible to establish the mechanisms and potential antiarrhythmic actions of pharmacological tools, safety. At the present time in the world practice the methods described above are practically not used.
Known methods based on surgical and/or mechanical disruption of the blood supply of the myocardium, aimed at identifying the antiarrhythmic activity. Small mammals, such as rats, verification of results using these approaches is extremely difficult, sleds is of a well-developed collateral circulation. When using large mammals experiments become extremely expensive, technical support experiments overly difficult.
Known methods of identifying pharmacological antiarrhythmic activity funds, which are based on electrical stimulation of the heart and enforce rhythm, alone or in combination with parasympathetic stimulation.
(Bhatt L. K., Nandakumar, K., S. L. Bodhankar Experimental animal models to induce cardiac arrhythmias. Ind. J. Pharm. 2005. 37. 348-357.)
These methods are effective to measure the ability to suppress persistent arrhythmia, but uninformative in terms of forecasting preventing ability and safety of antiarrhythmic drugs.
The closest way of the same purposes of the claimed invention is a method for predicting the efficacy and safety of antiarrhythmic drugs, which carry out the determination of the bioelectric parameters in isolated multicellular perfuziruemah preparations in vitro, representing sections of the working myocardium of the heart - the Atria, papillary muscles. This assess changes in the duration of action potentials (APS), refractoriness after application of the investigated compounds. Changing the duration of PD in the working myocardium is an element of regulation of cardiac activity in and of itself to them is no small predictive value: can talk about reducing, and increase the likelihood of arrhythmias.
(L. M. Hondeghem, L. Carlsson, G. Duker, Instability and triangulation of the action potential predict serious proarrhythmia, but action potential duration prolongation is antiarrhythmic. Circulation, 2001; 103, 2004-2013.)
The disadvantages of the above method are the low reliability and relevance with the actual pathophysiological processes that cause arrhythmias. You should also specify that this is a normal working myocardium of the heart and does not assess the effect of antiarrhythmic drugs on "natural arrhythmogenic substrate", which is the infarction pulmonary veins.
The present invention is to create an efficient method of pre-clinical studies cardiotropic antiarrhythmic drugs, increase reliability of predicting antiarrhythmic action, the security of potential pharmacological agents for this purpose, reducing the duration of the experimental phase and accelerates the transition to clinical research.
The technical result of the invention is to improve the reliability of predicting antiarrhythmic action potential pharmacological agents and the reduction in the duration of the experimental phase.
This is achieved by the fact that in the present method preclinical studies cardiotropic antiarrhythmic drugs, on the expectation definition bioelectric parameters in isolated multicellular perfuziruemah drugs and assessment of the change in the duration of action potentials according to the invention, as isolated multicellular perfuziruemah drugs used infarction pulmonary veins of the rats, and the changes of parameters are available in three modes multicellular preparations, additionally evaluate the resting potential and to increase DPD 90%, increase the ratio DRD 50%/DPD 90%, the decrease in the rate of spontaneous shift of the resting potential, reducing the most positive values of the membrane potential at rest the drug, to reduce repetition frequency packs spontaneous activity, reduce the frequency and variability of following the spontaneous PD in the stack, reducing the number and intensity reduction of postdeparture, the displacement of the membrane potential corresponding to the beginning of the burst of activity in the negative direction, assess signs of anti-arrhythmic action, and to reduce DPD 90% reduction relations DRD 50%/DPD 90%, increase in the rate of spontaneous shift of the resting potential, the increase in the most positive values of the membrane potential at rest the drug, increasing the repetition frequency packs spontaneous activity, increased frequency and variability of following the spontaneous PD in the stack, to increase the number and increase the intensity of postdeparture, the displacement of the membrane potential corresponding to the beginning of the burst activity in a positive way, appreciate the arrhythmogenic activity.
The technical result is achieved due to the use of patterns, with increased arrhythmogenic activity, namely infarction pulmonary veins of the rats.
Myocardial lining the pulmonary veins of the rats has a sufficient length, up to the bifurcations of the second order and intra-lungs plots, myocardial lining the pulmonary veins of the rats has properties characteristic of arrhythmogenic foci.
Myocardial lining the pulmonary veins Guinea pigs functionally indistinguishable from the atrial myocardium. Pulmonary vein of a rabbit do not have extensive myocardial plates, respectively, isolated preparations obtained from data mammal species, have low aritmogennosti and may not be used to evaluate the antiarrhythmic activity.
Use multicellular preparations, pulmonary veins, so as of great importance for the implementation of anti-arrhythmic action has a tissue level, factor interconnectedness of cardiomyocytes in tissue.
The implementation of the method
Multicellular preparations of pulmonary veins of the rats was obtained as follows: necrotizing animal, open chest, get the heart along with the lungs and placed in preparevalue bath (100 ml), filled with the perfusion solution at room temperature. Heart washed perfusion is astora (intraorale) containing heparin (or 0.2 units/ml). Ofpreparation the left atrium and pulmonary veins of the left lobe of the lung.
The pulmonary vein is separated from the proximal side in the area of the mouth, with the distal side in the area of the first bifurcation, cut along the longitudinal axis and deploy in a perfusion chamber, receiving the flat product (the area of drug ≈0.6-0.8 mm2). The drug deploy and fixed in the perfusion chamber endocardial side up. The procedure of preparation 5 minutes.
In Fig.1 shows a schematic representation of the preparation of the left atrium, pulmonary veins and the lobes of my lungs in rats, where SFM - the left atrial appendage, PL - free wall of the left atrium, LV - pulmonary vein, LDL - left lobe of the lung, DDL - incremental share of the lung. The dashed circled the area being examined.
After selection of a multicellular preparations are perfusion. Perfusion is performed at a temperature of 37°C and the velocity of the flow 15 ml/min (volume 3 cameras in 1 minute, which is necessary for adequate oxygenation of drugs). Perfusion solution continuously Oxygenium with Carbogen (95% O2and 5% CO2). Use of perfusion solution of standard composition: (mm): NaCl 133.47, KCl 4.69, NaH2PO4·2H2O 1.35, NaHCO316.31, MgSO4·7H2O 1.18, CaCl2·2H2O 2.5, glucose 7.77, pH of 7.2 to 7.4. Water mediterranei up to 1 litre.
Register potential generation is, action potentials. Action potentials away from "endocardial" side of the pulmonary veins. To register PD using glass microelectrodes filled with 3M KCl (resistance 1-30 Mω) connected to the amplifier. The amplified signal is fed to analog-to-digital Converter and a computer.
To predict the effectiveness and safety of potential antiarrhythmic drugs used perfusion of drugs pulmonary veins of the rats in three modes:
1. "Rhythmically excited medication"
The drug continuously stimulate rectangular pulses of 2 MS interval S=300 MS and an amplitude equal to two thresholds (1-3 B), which leads to occurrence of action potentials in the drugs. If necessary use other intervals of stimulation. Stimulating silver electrodes (D=0.5 mm) is placed in the proximal region of the drug.
Calculate the duration of PD at 50% and 90% repolarization (DRD 50%, DPD 90%) when the stimulation interval S=300 MS. Determine the increase in DPD 90% in the experimental groups compared with the DPD 90% in the control group.
2. "Resting drug"
After a 10-minute period of continuous stimulation and rhythmic activity cease stimulation. Termination of stimulation in the infarction pulmonary veins rats leads to spontaneous shift p of the building of peace in the region more positive values (Fig.2). Assess the degree and rate of spontaneous shift of the resting potential in the control groups, as well as after the action of the investigational pharmacological compounds.
In Fig.2 shows examples of action potentials and spontaneous shift of the resting potential in the myocardium of the pulmonary veins of the rats when the drug is in a resting state. 1, 2, 3 - ways to amend paragraphs at the termination of stimulation in the pulmonary veins (LV), 4 - atrial part of the drug (PL). The arrow indicates the moment of cessation of the stimulation.
3. "Spontaneously active drug"
The drug during the experiment perfusion 1 µm phenylephrine, resulting in infarction pulmonary vein of the rat develops a periodic train of spontaneous activity; spontaneous action potentials in packs related to early postdeposition.
In Fig.3 shows the cyclical nature of spontaneous burst activity and potential changes in the myocardium of the pulmonary veins of the rats under the action of phenylephrine: periods of spontaneous activity are accompanied by hyperpolarization, the rest period is accompanied by depolarization, where the bottom depicts a single PD preceding burst activity (0), the first DD in the stack, followed by early postdeposition (1 and 2), a typical train-PD (3), last DD tutu (67).
Assess the impact of the investigational pharmacological compounds on cha is Tautou repetition packs spontaneous activity, the most negative potential in the rest period, frequency, and variability of following the spontaneous PD in the stack, the intensity of postdeparture, the value of the membrane potential corresponding to the beginning of the burst activity.
Criteria for predicting the efficacy and safety of potential anti-arrhythmic and other pharmacological tools evaluation can serve the following bioelectric parameters that is possible only in the pulmonary veins of rats.
Namely, as signs of antiarrhythmic efficacy and safety are increasing DPD 90%, the increase of the ratio DRD 50%/DPD 90%, the decrease in the rate of spontaneous shift of the resting potential, the reduction in the most positive values of the membrane potential at rest the drug, reducing repetition frequency packs spontaneous activity, reduce the frequency and variability of following the spontaneous PD in the stack, reducing the number and a decrease in the intensity of postdeparture, the displacement of the membrane potential corresponding to the beginning of the burst of activity in the negative.
As features of arrhythmogenic activity are reduced DPD 90%, the decrease of the ratio of DPD 50%/DPD 90% of the increase in the rate of spontaneous shift of the resting potential, the increase in the most positive values of the membrane potential in oedema the drug, the increase in the repetition frequency packs spontaneous activity, an increase in the frequency and variability of following the spontaneous PD in the stack, increasing the number and the intensity of postdeparture, the displacement of the membrane potential corresponding to the beginning of the burst activity in a positive way.
Thus, the inventive method is effective, increases the reliability of predicting antiarrhythmic action, the security of potential pharmacological agents for this purpose reduces the duration of the experimental phase and accelerates the transition to clinical research.
The way preclinical studies cardiotropic antiarrhythmic drugs, including the definition of bioelectric parameters in isolated multicellular perfuziruemah drugs and assessment of the change in the duration of action potentials, characterized in that the isolated multicellular perfuziruemah drugs used infarction pulmonary veins rats, additionally evaluate the resting potential, and the changes of parameters are available in three modes multicellular preparations:
1) the drug is continuously stimulated by rectangular pulses of 2 MS interval S=300 MS and an amplitude equal to two thresholds (1-3), calculate the duration sweat the capacity of action at the level of 50% and 90% repolarization (DRD 50%, DPD 90%) when the stimulation interval S=300 MS, determine the increase in DPD 90% in the experimental groups compared with the DPD 90% in the control group;
2) after a 10-minute period of continuous stimulation and rhythmic activity cease stimulation, assess the degree and rate of spontaneous shift of the resting potential in the control groups, as well as after the action of the investigational pharmacological compounds;
3) the drug perfusion 1 µm phenylephrine for development infarction pulmonary veins rats periodic train of spontaneous activity, and assess the impact of the investigational pharmacological compounds on the repetition rate of the packets of spontaneous activity, the most negative potential in the rest period, frequency, and sequence variability of spontaneous action potentials (APS) in the stack, the intensity of postdeparture, the value of the membrane potential corresponding to the beginning of the burst activity;
to increase DPD%, increase the ratio DPD%/DPD%, the decrease in the rate of spontaneous shift of the resting potential, reducing the most positive values of the membrane potential at rest the drug, to reduce repetition frequency packs spontaneous activity, reduce the frequency and variability of following the spontaneous PD in the stack, reducing the number and intensity reduction of postdeparture, offset memb the data capacity, corresponding to the beginning of the burst activity in a negative way, appreciate the signs of anti-arrhythmic action, and to reduce DPD% reduction relations DPD%/DPD%, increase in the rate of spontaneous shift of the resting potential, the increase in the most positive values of the membrane potential at rest the drug, increasing the repetition frequency packs spontaneous activity, increased frequency and variability of following the spontaneous PD in the stack, to increase the number and increase the intensity of postdeparture, the displacement of the membrane potential corresponding to the beginning of the burst activity in a positive way, appreciate the arrhythmogenic activity.
FIELD: measurement equipment.
SUBSTANCE: invention comprises a method of determination of nano trace contaminants, supposing the use of emulsion from liquid crystal drops, dispersed in water and capable to change a configuration of liquid crystal drops in presence in the emulsion composition of foreign admixtures, measurement of change of intensity of light scattered by the emulsion, according to which it is possible to evaluate the content and density of required admixtures differing by that for a liquid crystal the compounds capable to trans-cis-transition under the effect of actinic light are selected and before measurement of change of light intensity the emulsion is illuminated additionally by actinic light, thus ensuring change of configuration in liquid crystal drops due to trans-cis-transition in liquid crystal molecules.
EFFECT: increase of sensitivity of method of determination of nano-trace contaminants.
SUBSTANCE: invention refers to a method for identifying living and dead mesozooplankton in seawater samples, which involves taking samples, staining the organisms with suitable colouring material, giving a visual estimation of the colour intensity of the units under the microscope, which is combined with microphotographying the units with an adjustable camera without changing the settings keeping throughout a photographic session of at least one sample; thereafter colour and brightness specifications average for each unit are measured in the formed images with using a painting program, e.g. Adobe Photoshop package, and the units are referred to living or dead by a discriminative analysis of the varied digital values.
EFFECT: improving the method.
SUBSTANCE: identification method of blue pus bacillus Pseudomonas aeruginosa involves inoculation of the test material on a hard substrate, incubation of the inoculum under anaerobic conditions at the temperature of 37°C during 16-18 hours. The obtained bacterial mass in the amount of one bacteriological loop is placed in 300 mcl of a physiological solution and warmed-up at the temperature of 98-99°C during 20-30 minutes and centrifuged at 12000 revolutions per minute during 30 seconds. To a supernatant there added is colouring material for electrophoretic detection in the quantity of 0.5 mcl and 20 mcl is added to a well with a size of 4×1 mm 1.2% agarose gel on TAE-buffer with 10 mcl of 1% ethidium bromide. Besides, the amount of 3 mcl of the solution of standard DNA-marker 1 kb containing DNA fragments in the range of 250-10000 bp is added to the test well. Horizontal electrophoresis is performed during 15-20 minutes, and at detection on the obtained electrophoregramme of the test bacterial mass of three strips radiating in ultraviolet light, one of which corresponds to fragments of standard DNA-marker with the size of 10000 bp, the second one corresponds to fragments of standard DNA-marker with the size of 6000-8000 bp and the third strip at the end of the track in the form of a whisk, which corresponds to fragments of standard DNA-marker with the size of less than 750 bp, there identified is Pseudomonas aeruginosa in the test bacterial mass.
EFFECT: method allows quick and complete identification of pigment-shaping and non-pigment strains Pseudomonas aeruginosa in a test bacterial mass.
5 dwg, 2 ex
SUBSTANCE: for analysis of removal with meadow grass of biochemicals the yield fluctuations are accounted depending on the structure of phytocenosis in the form of a ground cover. Conducting statistical data processing of testing the samples of grass from the test plots on the meander, central and terrace near flood plains, and also on the meadows with uneven and tiling location of grass species. And prior to establishment of sample plots the terrain reconnaissance is carried out with the grass cover selected for the measurements, an outline map of location of the components of the grass cover is made, then on each component of the grass cover in the form of a plot at least one temporary test plot is established, the wet and air-dry weight of the sample is determined by weighing on the cut grass sample. At that the tests on biochemical analysis are additionally carried out on the dried samples of grass to determine the concentration of at least three chemical nutrients in the form of mobile nitrogen, potassium and phosphorus oxides, and then by summing the concentrations of these three substances the total summarised removal of substances from the aerial part of grass on all test sites is calculated.
EFFECT: invention enables to determine the productivity of meadows.
9 cl, 12 dwg, 4 tbl, 1 ex
SUBSTANCE: for the purpose of the confirmation of a patient's death caused by ventricular fibrillation accompanying myocardial infarction in the patients died from myocardial infarction in the course of autopsy on the basis of a macroscopic pattern, the pathological process age is determined. This enables selecting examination areas in the heart at the level of necrosis. Absolute values (AV) of the following cell populations are counted in the x600-magnified visual field with using haematoxylin and eosin: necrosis-borderline lymphocytes (Lfb) in myocardial infarction of the age of 1-2 days, necrosis neutrophilic granulocytes (NGn) and fibroblasts (Fbn) in myocardial infarction of the age of 3-5 days. The derived absolute values are compared to threshold values (TV) with the threshold values of Lfb=2, NGn=38, Fbn=1. Each visual field is assigned with a diagnostic coefficient (DC), which is equal to 1.9 at the age of myocardial infarction of 1-2 days, if the absolute values Lfb<2 or -3.9, Lfb>2; if the age of myocardial infarction is 3-5 days, the diagnostic coefficient is -2.1, if AFbn<1, or the diagnostic coefficient is 7.2, if the absolute value of Fbn>1; the diagnostic value of 1.36 is shown by the absolute value of NGn<38; the diagnostic value is -11, if the absolute value is NGn>38. The total value (ΣDC) is calculated by moving between the visual fields within the examined area, and if ΣDC>12.78, the patient's death is stated to have come from ventricular fibrillation; if ΣDC<-12.78, the death is stated to be caused otherwise.
EFFECT: information value and reliability of the examination accompanying forensic medical examination.
1 tbl, 3 ex
SUBSTANCE: invention refers to laboratory diagnostics and can be used for testing biological samples for pathogenic microorganisms. The device for microbiological analysis of the body fluid samples comprises a compartment for incubating sample containers, an analyser for studying the internal atmosphere of the above containers and a sorting system for classifying the above containers in accordance with the carbon dioxide content detected by the above analyser. The sorting system classifies the containers delivered from the above incubation compartment to divide the above containers into the positive containers to be analysed for identifying microorganisms found in the respective samples, and the negative containers wherein the analysis is not required for identifying the microorganisms, and also the unspecified containers to be incubated once again.
EFFECT: invention provides simplifying and accelerating the analysis of the biological samples for the pathogenic microorganisms.
10 cl, 8 dwg
SUBSTANCE: method involves a post-therapeutic cytological examination of smears sampled from septic wounds, a cell composition analysis, including neutrophilic granulocytes and tissue elements - fibroblasts and fibrocytes; the material sampled from the septic wound on the 3rd day from the beginning of therapy is used for the cytological examination to determine accessory signs of therapeutic pathomorphism, namely the morphology of neutrophilic granulocytes, fibroblasts and fibrocytes and the presence of oxyphilic ground substance; observing the nuclear fragmentation and cytoplasm vacuolation in the neutrophilic granulocytes, multinuclearity, nuclear lobulation, karyonuclei in nuclei, basophilia, cytoplasm grain and vacuolation in fibroblasts and fibrocytes, as well as the presence of the oxyphilic ground substance, the treatment is stated to be effective.
EFFECT: invention provides objective clinical findings of the wound process even on the third post-therapeutic day and can be used for the reliable determination of the clinical effectiveness in septic wounds.
1 ex, 5 dwg
SUBSTANCE: invention refers to medicine. Substance of the early diagnostic technique for chronic renal disease consists in using a dosage range of dopamine of 1 to 3 mcg/kg of body weight and a standard water load of 200 ml. No glomerular filtration rate increase testifies to the presence of early signs of chronic renal disease.
EFFECT: using the declared technique enables standardising the examination as high as possible by precise dosage measurement of the preparation.
2 tbl, 2 ex
SUBSTANCE: carbon nanostructure surface is modified with (2,4,5-triiodophenyl)-methanol; the modified carbon nanostructures are analysed to measure a iodine amount; the prepared modified carbon nanostructures are administered into an experimental animal's body to take organs and tissues thereafter, to homogenise them in a 0.5-2 M NaOH solution, to sample a homogenate, to dilute with water, to expose the diluted sample to ultrasound to a temperature of 40-70°C, to determine a iodine amount in the prepared sample by mass spectrometry with inductively bound plasma and to calculate the carbon nanostructure content in the sample as shown by a difference of the iodine amount in the sample prior to administration of the modified carbon nanostructures and after administration thereof into the body and to re-calculate this iodine amount into the carbon nanostructure content in the sample with the use of the initial iodine content in the modified carbon nanostructure.
EFFECT: method provides monitoring in vivo of the carbon nanostructure distribution in the body.
2 cl, 6 dwg, 3 ex, 2 tbl
SUBSTANCE: invention refers to medicine, namely to oncohaematology, and can be used for prediction of the clinical effectiveness in the patients with non-Hodgkin lymphomas with bone marrow involvement. That is ensured by determining a lymphocyte volume and electric conductivity in patient's peripheral blood before the treatment and every next course of the chemotherapy with calculating a relation of the values. If observing a decrease of the relations as compared to the initial value, the positive clinical effectiveness and the absence of generalisation symptoms are predicted. The absence of the decrease or increase of the relations shows the absence or low clinical effectiveness and the presence of the symptoms of a continued tumour growth.
EFFECT: using of given method enables determining the sensitivity of the patients suffering non-Hodgkin lymphomas to the chemotherapy and performing the immediate and remote prediction of the clinical course of the disease.
1 tbl, 3 ex
SUBSTANCE: invention relates to method of determining platelet resistance to acetylsalicylic acid (ASA) by impedance analysis of aggregation function of platelets in vivo, in which aggregation activity is analyses after incubation of biological material sample with ASA with application of aggregation inducer, and aggregation of platelets is induced by collagen in optimal concentration 2 mg/ml, and simultaneously with measurement of impedance determination of dynamics of platelet granules release is performed by luminescent method, where before carrying out aggregation samples are calibrated by means of adenosine triphosphate (ATP) standard, value of aggregation amplitude are determined in Ohms by obtained aggregatograms and obtained values are given points: values ≤6 correspond to 0 points, values 7-9 correspond to 1 point, values 10-12 correspond to 2 points, values >12 correspond to 3 points; after that, intensity of ATP release from platelet granules is determined in nmoles and obtained values are given points: values <0.5 correspond to 0 points, values 0.5-1.0 correspond to 1 point, values 1.0-1.5 correspond to 2 points, values 1.5 correspond to 3 points, and then resistance index (RI) is calculated by formula, value of IR parameter points to presence of aspirin-resistance of platelets.
EFFECT: carrying out fast complex diagnostics of platelet resistance to ASA with estimation of functional state of platelet granules before beginning treatment and possibility of preventing development of undesirable thrombotic or hemorrhagic complications.
2 dwg, 1 tbl, 2 ex
SUBSTANCE: claimed is application of fat emulsion for parenteral feeding as solvent for compounds which are poorly soluble in water. Fat emulsion contains in 1 l of solution: 30 g of refined soybean oil, 30 g of triglycerides with the average chain length, 25 g of olive refined oil, 15 g of purified fish oil.
EFFECT: obtaining solvent for compounds, poorly soluble in water, which makes it possible to determine parameters and spectrum of biological activity of novel compounds of chemical nature at the stages of pre-clinical and clinical tests, which does not change basic biological constants and possesses biological inertness.
2 tbl, 2 ex
SUBSTANCE: invention relates to biology, forensic and analytical chemistry and specifically to methods of determining procaine in blood plasma. The method comprises adding sodium fluoride to blood plasma containing procaine to achieve concentration of 10 mg/ml; treating the obtained mixture with acetone; separating the extract from the precipitate by filtering; evaporating acetone from the filtrate in an air current at room temperature; diluting the aqueous residue by adding water; saturating the obtained solution with ammonium sulphate; alkalising with an ammonium buffer solution to pH 9.0-9.5; extracting twice with portions of an organic extraction agent in the form of 30% camphor solution in methyl acetate, with ratio of the aqueous to the organic phase of 1:1 by volume; separating the organic extracts; combining; evaporating the solvent from the combined extract in an air current at room temperature; chromatographing the residue in a thin layer of silica gel STKH-1A on Sorbfil PTSKH-AF-A-UF plates, using a dichloromethane-ethanol mobile phase in ratio of 6:4 by volume; developing the chromatogram in UV light; eluting the analysed substance from the sorbent with a mixture of acetonitrile-methanol-0.025 M potassium dihydrogen phosphate solution with pH 3.0 in ratio of 10:10:90 by volume; chromatographing by HPLC method using a reversed-phase sorbent Nucleosil C18, a polar mobile phase acetonitrile-methanol-0.025 M potassium dihydrogen phosphate solution with pH 3.0 in ratio of 10:10:90 by volume and a UV detector; measuring optical density at wavelength 298 nm and calculating the amount of the analysed compound from the area of the chromatographic peak.
EFFECT: method improves sensitivity of determination.
3 tbl, 2 ex
SUBSTANCE: invention relates to analytical chemistry. The method is characterised by electrochemically concentrating benzoic acid on the surface of a graphite electrode for 90 s at electrolysis potential of (-0.500) V on a background of 0.1 mol/l sodium hydrogen phosphate, recording polarisation curves with linear potential sweep rate of 25 mV/s and determining concentration of benzoic acid from the peak height in potential range of 0.5-1.6 V relative to a silver chloride electrode.
EFFECT: method provides highly sensitive and rapid determination of benzoic acid in medicinal drugs.
3 ex, 6 tbl
SUBSTANCE: sample is applied on a paper filter, and standard calibration solutions of metronidazole in the concentration range of 10-100 mcl are radially applied on the same filter. The filter is processed with a developing agent containing 5% potassium hydroxide and acetone taken in ratio 2:1 and thermostated at 100°C until coloured spots are visible. A colour intensity of the sample spot is compared to the colour of the spot colour of the reference calibration solutions to determine the metronidazole concentration in the analysed sample.
EFFECT: method enables the fast and reliable monitoring of the metronidazole content and the timely correction of the therapeutic process.
SUBSTANCE: invention refers to medicine, namely to studying and analysing medical preparations, and can be used for standardising herbal raw materials. A method for identification and qualitative measurement of chlorophyll, carotinoids and hydroxycinnamic acids in a combination in great nettle leaves involves a 1-hour fractional extraction, 30 min each of the ground raw material having a particle size of 1.0 mm on a water bath at a temperature of 100°C with 70% ethanol in a ratio of the herbal raw material to the extractant of 1:100, the combination of the extracts and reduction to 100 ml with a solvent, dilution of the prepared solution in a ratio of 2:25 in 96% ethanol, measuring an optical density of the solution in relation to 96% ethanol at maximum absorption 328±1 nm, 442±1 nm and 667±1 nm, calculation of the total content of hydroxycinnamic acids equivalent to chlorogenic acid, carotinoids equivalent to violaxanthin and chlorophyll in the percentage equivalent to an absolute dry mass of the raw material by formulas.
EFFECT: method provides availability, simplicity, efficiency and low error of measurement.
3 dwg, 1 ex
SUBSTANCE: method of determining fat-soluble vitamins A, D2, E and β-carotene which are present at the same time includes separating the fat-soluble vitamins from a substance by extraction with 96% ethanol, separating the alcohol extract of vitamins using a separating funnel, successive chromatography using Sorbfil PTSKH-P-A silica gel plates on a polymer substrate using two eluents with a different range, time of saturating the chamber with eluent vapour of 20 minutes and elution time of 55 min; drying the plates at temperature not lower than 80°C in a temperature-controlled chamber for 3-5 min, treating the plates with a developer - 5% alcohol solution of phosphatomolybdic acid; according to the invention, the eluents used are hexane:chloroform (19:1) and hexane:chloroform (3:1), and detection of the chromatographic zone of β-carotene is carried out before treating the plates with a developer in day light.
EFFECT: simpler and faster process of determining fat-soluble vitamins.
10 dwg, 3 ex
SUBSTANCE: invention relates to medicinal drug quality control and provides a method of authenticating and determining the quantitative content of benzethonium chloride in medicinal drugs, which includes separating components of the drug via high-performance liquid chromatography, where the eluent used is a mixture of acetonitrile, tetrabutylammonium hydrophosphate, disubstituted potassium phosphate and water, with content of tetrabutylammonium hydrophosphate of 2.5 mM, content of disubstituted potassium phosphate of 5 mM, wherein separation of the components of the drug is carried out with column length of 125 mm.
EFFECT: high accuracy and faster analysis since there is no need for special sample preparation.
SUBSTANCE: invention relates to a method of screening a non-teratogenic substance comprising bringing a test substance into contact with cereblon or a fragment of cereblon, evaluating the capacity of the test substance to bind to cereblon or the fragment of cereblon, and selecting a test substance that does not bind to cereblon or the fragment of cereblon or a test substance exhibiting lower capacity to bind to cereblon or the fragment of cereblon compared with thalidomide.
EFFECT: improved method.
8 cl, 19 dwg, 8 ex
SUBSTANCE: method involves taking blood at 1 ml per a sample into the Vakutainer evacuated test tube with sprayed ethylene diaminetetraacetic acid 2.2 mg; a reference sample 1.0 ml and a sample 1.0 ml with weighted xenobiotics are taken from the total volume into the Eppendorf tubes; 30-minute incubation is followed by using a uniform procedure to prepare blood smears by applying on a dry slide to prepare a smear to be stained according to Leishman; blood corpuscles are analysed by the Zeiss light microscope at a magnification 1500. A microscope field showing echinocytes with styloid processes of the membrane not found in the reference sample is a sign of the xenobiotic membrane toxicity.
EFFECT: invention enables an instant assessment of the membrane toxicity of the substance.
SUBSTANCE: polysomnography is conducted. Slow sleep phases (SSP) and fast sleep phases (FSP) are determined. A maturity index of integrative sleeping apparatuses (MIS) is calculated by formula MIS = SSP/FSP. If the MIS is less than 1.5, a physiologically optimum structure of the nocturnal sleep is stated in a healthy child.
EFFECT: method enables assessing the quality of the nocturnal sleep in the children.
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