Method of determining biological inequivalence of nanodiamonds
SUBSTANCE: incubation medium, oxidation substrate and tetraphenylphpsphonium chloride as indicator are placed into respective measuring cell of installation for measuring mitochondria potential, provided with tetraphenylphosphonium-selective electrode, change of tetraphenylphosphonium concentration is registered and when constant tetraphenylphosphonium concentration is achieved, mitochondria, isolated from animal organism, are added. Change of membrane potential of mitochondria is registered by change of electrode signal, when constant potential is achieved, respective water suspensions of analysed samples of nanodiamonds with pH 7.2-7.4 are added, and value of rate of change of mitochondria membrane potential is measured. Presence of statistically reliable difference of rates of mitochondria membrane potential change testifies to biological inequivalence of compared samples of nanodiamonds.
EFFECT: expressive and available method of determining biological inequivalence of nanodiamonds.
2 cl, 1 tbl, 1 dwg, 1 ex
The present invention relates to the field of pharmacology, biopharmaceutics and pharmaceutical industry and relates to a method of determining the biological neweventnotify samples of nanodiamonds, which can find application in the quality control of industrially produced and modified nanodiamonds, as well as in the production of pharmaceuticals and drug delivery based on the nd.
Nanodiamonds are a distinct group of carbon nanoparticles. A special place among them is occupied by detonation nanodiamonds, which are obtained by detonation of explosives or their mixtures in a special chamber. As a result of detonation is formed mixture, the composition of which comprises particles of nd. To highlight nd the mixture is subjected to oxidation treatment in harsh environments .
Received detonation nanodiamond has a number of distinctive physical and chemical characteristics: ultra-small size of the primary particles (4-6 nm), highly developed specific surface (up to 400 m2/g), high concentration of functional groups therein, and chemically inert core particles .
There are several technology options for isolation and purification of detonation nanodiamond, each of which affects the physico-chemical characteristics of the resulting particles and can affect their biological properties. Cu is IU, it is known that further chemical modification of nanodiamond also affect its biological properties.
So, in  studied the toxicity of samples of nanodiamonds brand "ultraFine Diamond" (Russia) and manufactured by General Electric (USA) on-line lung epithelium adenocarcinoma human A. Nano-diamond mark "ultraFine Diamond" (Russia) obtained by the method of detonation synthesis, and the final stage of cleaning can vary from treatment with perchloric acid, potassium permanganate prior to cleaning with nitric acid . Therefore, the final properties of the brand of nd brand "ultraFine Diamond", depending on the stage of its final treatment, can be different . Nanodiamond manufactured by General Electric (USA) obtained by the method of static synthesis with subsequent acid treatment . If nd made in the USA of cell survival was higher than that of nd, produced in Russia. In turn, the modification (oxidation of the mixture of acids) samples nd these two brands leads to a decrease of cell survival .
In  was used nanodiamonds obtained in the NanoCarbon Research Institute Co., Ltd (Japan). Technology data of nanodiamonds includes oxidation process, and then grind nanodiamond powder in a ball mill using Zirconia balls . It is shown that the survival of the notches in the case of processing the nanodiamond is 80-90% compared with control. Subsequent surface modification of nanodiamond polyethylene glycol reduces the survival rate of cells to 70-80% and its amination of up to 55% .
Mandatory application of nanodiamond in medicine as carrier systems for drug delivery is the identity of its chemical and physico-chemical and pharmacotoxicological characteristics. For the nd in biomedical applications of the above works [2-6] clearly implies the need to find and develop ways of controlling non-bioequivalent nanodiamonds obtained by different technologies and undergo different chemical treatment, including chemical modification.
Currently in the patent and scientific literature does not describe how to identify differences in biological effects and biological neweventnotify samples of carbon nanoparticles, including nanodiamonds obtained using various techniques and/or different ways of chemically modified.
The aim of the invention is to develop a rapid and affordable method for determining the biological neweventnotify samples nd.
The inventive method consists in the comparative determination of the effect of samples nd on the membrane potential of mitochondria of rat liver. The essence of what the procedure is the following. In the measuring cell make a mixture of freshly isolated rat liver mitochondria, substrate oxidation and the indicator are added, and the suspension of nd and record the rate of change of the concentration of free indicator, which determines the change in membrane potential of mitochondria. On the basis of comparison of velocity changes of membrane potentials, obtained by the action of the analyzed and reference materials, to judge the presence or absence of biological neweventnotify these samples. As a standard sample (reference sample) can be selected by any of the studied sample nd depending on the purpose of its further use.
Mitochondria are the Central link in the chain of energy for cellular processes, which in the presence of substrate oxidation (e.g., succinate sodium/potassium pyruvate sodium/potassium, and others) actively generate the membrane potential inside the membrane . To determine the membrane potential use indicators that respond to changes in its size. As these indicators can serve a variety of hydrophobic ions (tetraphenylphosphonium, tetraphenylboron) and various probes that are sensitive to surface and transmembrane potential (for example, rhodamine-123, oxonol-6, N,N-dimethyl-N-nonyl-N-temperaturebased and others) . Known is about, that link membrane potential of mitochondria (Δφ) with the concentration of free (recordable) form of the indicator (Coutand concentration of the indicator inside the mitochondria (Cin) is described by the Nernst equation :
where R is the universal gas constant, equal 8,31 j/(mol·K);
T is the absolute temperature (K);
z is the charge of an ion (for tetraphenylporphine z=1);
F - Faraday constant, equal 96485,35 KL/mol.
From the Nernst equation, it follows that with increasing membrane potential indicator, for example tetraphenylphosphonium, begins to penetrate into the mitochondria, the concentration of the free form in the cell is reduced. With decreasing membrane potential, on the contrary, there is a release indicator from the internal space of the mitochondria, thereby increasing the amount of free indicator.
To assess the impact of nd on the membrane potential of mitochondria we propose to use the value not the membrane potential, and the speed of its change, which is measured by changing the value of the concentration of free indicator in the cell per unit of time (min.)
The inventive method are described in more detail in the following. To compare samples of nd, taken in an amount of 0.1-0.25 mg add 2 ml of distilled water. Adding the Astor potassium hydroxide bring the pH of the suspension to a value of 7.2 to 7.4. The measurements were carried out using potentiometric computerized installation tetraphenylphosphonium-selective electrode . In the measuring cell is placed 20 μl of mitochondria isolated from liver homogenate of rats according to standard methods , which is that of mitochondria isolated from the liver of adult male Wistar rats in accordance with standard procedure, including differential centrifugation and storage of selected mitochondrial ice .
Incubation of mitochondria in all samples is carried out in a standard environment: 125 mm potassium chloride, 15 mm 4-(2-oxyethyl)1-piperazineethanesulfonic acid, 1,5 mm phosphate at pH of 7.25 in the presence of 4 mm potassium succinate. Add indicator (up to a concentration of 1 μm). Then add an aliquot (10-50 µl) of the suspension sample nd and record the rate of change of the concentration of free indicator. According to the measurement results build graphs of the effect of the compared samples of nd on the membrane potential in the coordinate concentration of free indicator - time” (Fig.1).
To add to the cell mitochondria suspensions of the samples of the inherent nature of all curves practically do not differ. It is significant curves after adding to the mitochondria of samples nd (in Esenia which is marked in Fig.1 by arrows). Choose the linear portion of the curve after addition of the sample and calculate the change in the concentration tetraphenylporphine in time, expressing the result as the rate of change in concentration of the indicator in a minute.
On the basis of statistically significant comparisons (±0,025 - the measurement of these quantities make a conclusion about the presence or absence of biological neweventnotify compared samples of nd.
A brief description of graphic materials.
Fig.1. Decline curves membrane potential of mitochondria under the influence of industrial samples and modified samples nd (time additive samples of nd to the mitochondria marked by arrows). 1 - control; 2 - hydrogenated nanodiamond brand "UDA-TAN; 3 - nano-diamond mark "UDA-TAN; 4 - nano-diamond mark "Standard ND"; 5 - chlorinated nano-diamond mark "UDA-TAN".
The invention is illustrated by the following example.
According to the claimed method was analyzed 4 sample nanodiamonds, including two samples of industrial nd: nd brand "UDA-TAN" (SKTB "Technologist", Russia) and nano-diamond mark "ND Standart" ("Adamas Nanotechnologies, USA) (samples 3 and 4, respectively). Nano-diamond mark “UDA-TAN obtained by detonation synthesis with stage cleaning 50-60% nitric acid at a temperature of 230-240°C and a pressure of 80-90 ATM and followed what ammonolysis under pressure at 200°C (pH=10) [1, page 76-108].
Details of the production technology nd brand "ND Standart" is unknown.
Nanodiamonds with gidrirovannoe (sample 2) and chlorinated (sample 5) surfaces were obtained by chemical modification of the sample 3 in accordance with the methods of .
Mitochondria isolated from the liver of adult male Wistar rats in accordance with the standard procedure by differential centrifugation . All requirements for the care and working with animals were carefully observed. Liver was homogenized in ice buffer containing 70 mm sucrose, 10 mm 2-amino-2-hydroxymethyl-propane-1,3-diol and 1 mm ethylene glycol-bis(2-aminoethylamide ether)-N,N,N,N-tetraoxane acid (pH of 7.4). The homogenate was centrifuged at 600 g for 7 minutes at 4°C, were selected fraction of the supernatant and centrifuged at 9000 g for 10 min to sediment the mitochondria. Mitochondria were twice washed in the above medium without ethylene glycol-bis(2-aminoethylamide ether)-N,N,N,N-tetraoxane acid and centrifuged. Next, the precipitate of mitochondria containing 60 mg of protein, suspended in wash medium and kept on ice.
As a control, we measured the membrane potential of mitochondria without adding nanodiamonds (curve 1 in Fig.1). To compare samples of nd, each of which took in an amount of 0.1 mg, EXT is ulali 2 ml of distilled water, thoroughly mixed, the mixture and adding a solution of potassium hydroxide brought the pH of the resulting suspension to a value of 7.4.
For analysis of each sample in the cell was placed 20 ml of mitochondria, isolated by the method of . An indicator used chloride solution tetraphenylporphine, which added to the value of its concentration in the mixture equal to 1 μm. Further, as the substrate oxidation used succinate potassium and record the concentration of free tetraphenylporphine in the measuring cell. Then for each sample inherent in the measuring cell was added 30 μl of the previously prepared suspension and recorded the change in the concentration tetraphenylporphine on the linear parts of the curves after addition of the samples of nd (Fig.1). Next chose the linear region of the curve after addition of the sample, and calculated the change in the concentration tetraphenylporphine in time, expressing the result as the rate of change in concentration of the indicator in a minute. The measurement was repeated in strictly identical conditions not less than three times. Based on the obtained values of the rate of change of concentration tetraphenylporphine for each sample to calculate the average value and the magnitude of the statistical error. On the basis of statistically significant differences in the velocities is modify the concentration of the indicator for different samples nd did the conclusion of their biological neweventnotify. The results of the measurements are shown in the Table.
As the comparison sample used sample nd Russian production (sample 3).
|The results of the experiment to determine bioequivalence samples nd () (relative to the sample 3) of the claimed method.|
|No. sample||Sample||The rate of change of concentration of the indicator, µm/min||Bioequivalents|
|2||Hydrogenated nanodiamond brand "UDA-TANG"||0,025±0,025||No|
|3||Nano-diamond mark "UDA-TAN" (reference sample)||0,035±0,025||-|
|4||Nano-diamond mark "Standard ND"||0,377±0,025|
|5||Chlorinated nano-diamond mark "UDA-TANG"||0,284±0,025|
From the Table it follows that the speed of decrease of the membrane potential in the event of exposure of samples of industrial nd Russian production and nd with gidrirovannoe surface is 0,035±0.025 and 0,025±0,025 µm/min, respectively. As the speed difference not statistically significant, it was concluded on biological equivalence of samples 2 and 3 (i.e., about the lack of bioequivalence). At the same time, the speed of decrease of the membrane potential for samples 4 and 5 is 0,377±0.025 and 0,284±0,025 µm/min, respectively, which, given the statistically significant differences in the velocities, allows to make a conclusion about their biological neweventnotify not only among themselves but also with respect to other samples of nanodiamonds.
1. C. Y. Dolmatov. Detonation nanodiamonds. Production, properties, application. / St. Petersburg: Izd. NGO "Professional", 2011. 536 S.
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3. F. Huang, Yi Tong, Sh. Yun. Synthesis Mechanism and Technology of Ultrafine Diamond from Detonation // Physics of the solid body. 2004. So 46. No. 4. S. 601-604.
4. C. B. Muratov, A. A. Vasiliev, V. C. Matias, I. I. Duda. The influence of gas-forming impurities on the heat capacity of Nanoka the metallic diamond detonation synthesis // Nanostructured materials. 2011. No. 1. S. 23-31.
5. X.-Q. Zhang, M. Chen, R. Lam, X. Xu, E. Osawa, D. Ho. Polymer-Functionalized Nanodiamond Platforms as Vehicles for Gene Delivery // ACS Nano. 2009. V. 3. N. 9. P. 2609-2616.
6. E. Osawa. Remarks on the handling of colloidal solutions of 5-nm diamond particles / NCRI Technical Bulletin, 2009. No..3. P. 1-8.
7. D. Nelson, M. Cox. Fundamentals of biochemistry Lehninger: 3 so So 2: Bioenergetics and metabolism. / Lane. from English. - M.: BINOM. Knowledge laboratory. 2014. S. 306-342.
8. N. Hennis. Biomembranes. Molecular structure and function. / M.: Mir. 1997. S. 319-321.
9. N. Kamo, M. Muratsugu, R. Hongoh, and Y. Kobatake. Membrane potential of mitochondria measured with an electrode sensitive to tetraphenyl phosphonium and relationship between proton electrochemical potential and phosphorylation potential in steady state // J. Membrane Biol. 1979. V. 49. P. 105-121.
10. N. And. Paducheva In. A. Teplova, N. In. Beloborodov. Role of thiol antioxidants in the restoration of mitochondrial function, the modified microbial metabolites. // Biophysics. 2012. So 57. No. 5. S. 820-826.
11. N. I. Fedotcheva, R. E. Kazakova, M. N. Kondrashova, N. V. Beloborodova. Toxic effects of microbial phenolic acids on the functions of the mitochondria // Toxicology Letters. 2008. V. 180. P. 182-188.
12. G. V. Lisichkin, I. I. Kulakova, A. Yu. Gerasimov, A. V. Karpukhin, R. Yakovlev Yu. Halogenation of detonation-synthesis nanodiamond surface // Mendeleev Communication. 2009. No. 19. P. 309-310.
1. The method of determining the biological neweventnotify of nanodiamonds by comparative determine the effect of sample nd on the membrane potential of the mitochondria of animals, namely, that corresponding to the measuring cell unit for measuring the capacity of the mitochondria, is equipped with tetrafen lovoni-selective electrode, put the incubation medium containing an aqueous solution of potassium chloride, 4-(2-oxyethyl)1-piperazineethanesulfonic acid, phosphate buffer, substrate oxidation and as an indicator of chloride tetraphenylporphine, register the change of concentration tetraphenylporphine and when constant concentration tetraphenylporphine add isolated from animals mitochondria change signal electrode register the change of membrane potential of mitochondria, while achieving constant potential add appropriate water suspensions of the samples of nanodiamonds with a pH of 7.2-7.4 and measure the magnitude of the rate of change of membrane potential of mitochondria, the presence of statistically significant differences in the speed change of the membrane potential of mitochondria indicates biological neweventnotify compared samples of nanodiamonds.
2. The method according to p. 1, where the substrate oxidation using potassium succinate.
SUBSTANCE: hose of an artificial air feed apparatus is inserted into a lobar bronchus of the estimated injured lobe of the lung. The hose is fixed by ligaturing the bronchus. The air is pumped into the examined lobe of the collapsed lung until it extends completely, and a microinjury is visualised.
EFFECT: technique can provides the more reliable diagnosis of the lung microinjury improve in dead bodies, which is achieved by filling the collapsed lung with air providing opening the microinjury.
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 refers to medicine, namely to hepatology, and describes a method for examining the liver detoxification function state. The method involves measuring fasting blood AWM (average weigh molecule), ALT (alanine aminotransferase) and AST (aspartate aminotransferase), measuring the same values the night before at bedtime, as well as measuring a circulating blood volume and a packed cell volume in the morning and in the evening followed by determining a coefficient of the liver detoxification function state; and if observing the derived coefficient K≤0.6, the liver detoxification function state is considered to be satisfactory, while the coefficient K>0.6 shows the depressed liver detoxification function.
EFFECT: method provides the more reliable assessment of the liver detoxification function state.
SUBSTANCE: method involves the preliminary sorption of a specific ligand in polystyrene tray wells; the above ligand is specified in: haemoglobin, myoglobin, collagen, fibrinogen, fibronectin, immunoglobulin A; that is followed by adding a microorganism cell suspension into the tray wells with the sorbed ligand, incubating the cell suspension in the tray wells for 15 minutes, sampling a suspension aliquot from the well and adding it to wells of another or the same tray containing 0.5% sodium chloride or 0.2 M sodium phosphate; the bacterial cell adhesion is assessed by measuring a decrease of the optical density of the prepared diluted suspensions at wave length 600 nm as compared to the well references free from ligands.
EFFECT: invention is characterised by a high test rate of the microorganism adhesion to ligands of various nature, high reproducibility, using a minimum amount of the microorganism biomass, with no need for hazardous chemicals to be used.
7 tbl, 7 ex
SUBSTANCE: invention relates to biology and toxicological chemistry and can be used in practice of sanitary and epidemiological stations, chemical-toxicological, forensic and veterinary laboratories. A biological material, containing substituted 2-methoxyhydroxybenzene, is two times (each time for 30 minutes) infused with ethylacetate with mixing, separate extracts are separated from solid particles of the biological material, combined, ethylacetate is evaporated in an air flow at 18-22°C, residue is repeatedly processed with acetone, acetone extracts are separated, combined, dehydrated, evaporated in an air flow at 18-22°C, and then in a nitrogen flow until a solvent is completely removed, residue is dissolved in hexane, extracted with a buffer solution with pH 12-13, a water-alkaline extraction is separated, acidified to pH 2-3, saturated with sodium sulphate, extracted with diethyl ether, the ether extract is separated, dehydrated, evaporated in an air flow at 18-22°C, and then - in a nitrogen flow until the solvent is completely removed, residue is dissolved in a mixture of solvents hexane-dioxane-propanol-2, taken in a ratio of 20:5:1 by volume, chromatographed in macrocolumn with silicagel KSS No 3 80/120 mcm with the application of a mobile phase hexane-dioxane-propanol-2 in a ratio of 20:5:1 by volume, eluate fractions, containing the analysed substance, are combined, the eluent is evaporated first in an air flow at a temperature of 18-22°C, and then in a nitrogen flow until the solvent is completely removed, residue is dissolved in dichloromethane, processed for 20 minutes with N-tert-butyl-dimethylsilyl-N-methyltrifluoroacetamide under conditions of heating at a temperature of 60°C with carrying out determination by a chromatography-mass spectrometry method with the application of a capillary column 25 m long with an internal diameter of 0.2 mm with an immobile phase (5%-phenyl)-methylpolysiloxane, with the application of a mass-selective detector, working in an electron impact mode, an initial temperature of the column thermostat constitutes 70°C, the said temperature is kept for 3 minutes, further the temperature is programmed from 70°C to 290°C at a rate of 20°C per minute, the final column temperature is kept for 10 minutes, the injector temperature constitutes 250°C, the quadrupole temperature is 150°C, the temperature of a ion source is 230°C, the temperature of the detector interface is 300°C, intensity of a signal, conditioned by charged particles, formed in the bombardment of the analysed substance, leaving the capillary column and getting into the ion source, with a ionising beam of electrons with the energy of 70 eV, is registered, the mass-spectrum by the complete ion flow is registered and an amount of substituted 2-methoxyhydroxybenzene is calculated by the area of a chromatographic peak of its trimethylsilyl derivative.
EFFECT: achievement of an increased analysis sensitivity.
4 tbl, 3 ex
SUBSTANCE: invention relates to biology and toxicological chemistry and can be used in chemical-toxicology, expert forensic and clinical laboratories. The method includes: crushing a biological object containing N-(4-nitro-2-phenoxyphenyl)-methanesulphonamide, settling twice for 45 minutes with portions of an organic isolating agent which is methyl acetate, combining the obtained extracts, evaporating the solvent from the resultant extract, treating the residue with acetone, separating the acetone extract, evaporating the solvent from resultant extract, dissolving the residue in diethyl ether, extracting the ether solution with a buffer solution at pH 9-10, acidifying the aqueous alkaline extract with 24% hydrochloric acid to pH 2-3, saturating the obtained solution with sodium bromide, extracting with ethyl acetate, evaporating the obtained extract in an air current at 20-22°C until a dry residue is obtained, dissolving the residue in a mixture of hexane and acetone taken in volume ratio of 8:2, performing chromatography on a macrocolumn with silica gel L 40/100 mcm using a hexane-acetone mobile phase in volume ratio of 8:2, combining eluate fractions containing the analysed substance, evaporating the eluent in an air current at 20-22°C until complete removal of the solvent, dissolving the residue in methanol and performing determination via a combined physical-chemical method in the form of chromatography-mass spectrometry, using a DB-5 MS EVIDEX capillary column with a mobile phase which is 5% phenyl-95% methylpolysiloxane, using a mass-selective detector operating in electron impact mode, the initial thermostat temperature of the column is 70°C, maintaining said temperature for 3 minutes, further raising the temperature from 70°C to 290°C at a rate of 20°C per minute, maintaining the final temperature of the column for 16 minutes, the temperature of the injector is 250°C, the temperature of the quadrupole is 150°C, the temperature of the detector interface is 300°C, detecting strength of the signal resulting from charged particles formed when bombarding the analysed substance coming from the capillary column and falling into an ion source with an ionising electron beam with energy of 70 eV, recording the mass spectrum on the full ion current, while calculating the amount of N-(4-nitro-2-phenoxyphenyl)-methanesulphonamide from the area of the chromatographic peak.
EFFECT: high sensitivity of analysis.
2 ex, 3 tbl
SUBSTANCE: invention relates to medicine, namely to clinical, laboratory diagnostics, microbiological methods of research, and is aimed at standardisation of saliva analysis by method of wedge dehydration/crystallography. Claimed is method of obtaining standard, quality sample of mixed saliva facia for crystallography with application of portable laboratory device with inbuilt levels on axes X and Y, legs, regulated by height and isolating cover. From 20 to 40 microscope slides are simultaneously placed on the surface of portable laboratory device after obtaining smooth horizontal without inclination angle surface. After that, one sample of mixed saliva in amount 0.02 ml, preliminarily centrifuged for 20 min at 3000 rev/min, is applied on each microscope slide by means of micropipette. As a result a round 1.0 mm high drop of mixed saliva with diameter from 4.0 to 5.0 mm, corresponding to standard parameters, is obtained. Then, standard in dimensions drop on microscope slide placed on the surface of portable laboratory device, adjusted by levels, is dried at room temperature +18…+25°C under isolating cover for 5 hours, with further performance of microscopy of standard quality sample of saliva facia.
EFFECT: obtained sample makes it possible to interpret indices of crystallography without distortions, as standard in volume, dimensions and shape drop of mixed saliva is obtained, and there is no displacement of crystallisation centre and impairment of figures of saliva facia (crystallography pattern) in drop, dried on ideal horizontal surface of the device, ratio of central and peripheral zones of facia exactly correspond to organism's condition, which reduces percentage of false results of crystallography.
3 ex, 1 tbl, 3 dwg
SUBSTANCE: invention aims at asserting the maximum allowable blood concentrations (MAC) of heavy metals in the children living in the dirty environment as shown by health risk criteria after the chronic integrated exposure. An environmentally neglected zone is selected; a representative sampling of the children for the examination is drawn that is a basic group with using biological, social and hygienic criteria; the same criteria are used to draw a representative sampling of the children to a reference group living in the environmentally friendly zone. In the territory of the above zones, the chronic exposure of the analysed heavy metal is qualitatively assessed by establishing its average daily concentration in the ambient environment; the derived value is used to calculate a total average daily doses of a heavy metal supplied from various sources into a child's body averaged over the annual exposure for the children of both groups. Blood is sampled from the children every three months for one year to determine the content of the analysed heavy metal and also to measure the biochemical values of blood plasma and serum characterizing body responses presented by actual or potential health problems that are response markers. That is followed by calculating the average blood concentration of the analysed heavy metal and comparing it to the reference for the same heavy metal with using a Student two-sample test, thereby stating whether the children were sampled from the main and reference groups adequately. A mathematical modelling procedure is used to establish a relation between the exposure that is the total average daily doses of the analysed metal, and the exposure marker that is the average blood metal concentration. A sliding window technology is used to assert the response markers selected. The maximum allowable concentration of the exposure marker and respective marker is determined by a technique based on ratio analysis.
EFFECT: enabled measurement of the blood MAC of the heavy metals in the children after the integrated exposure with using sparing techniques making it possible to avoid a health risk.
4 tbl, 2 dwg, 1 ex
SUBSTANCE: invention represents an instant diagnostic technique for acute intestinal infections (AIIs), involving detecting indication markers of the AII aetiology with the use of laboratory immunology tests, differing by the fact that the AII aetiology is stated in children of an early age category, preferentially in the newborn children; that is accompanied by measuring the concentration of cytokine, interleukin IL-10 in coprofiltrate and diagnosing chronic placental insufficiency (CPI); a probability (P) of the bacterial AII aetiology is calculated; the value P of more than 50% testifies to the bacterial AII aetiology, while the value P being less than 50% shows the absence of the bacterial AII aetiology, and enables considering the diagnostics second stage to be necessary, which implies measuring the concentration of cytokine, interleukin IL-4 in coprofiltrate; the time of latching the newborn child to the breast is established with considering the type of feeding; that is combined with calculating a probability (P) of the viral or viral-bacterial AII aetiology, with the value P of more than 50% testifying to the viral AII aetiology, while the value being less than 50% makes it possible to state the viral-bacterial AII aetiology.
EFFECT: more accurate diagnosing of the aetiology of acute intestinal infection and simplifying the diagnostic procedure.
SUBSTANCE: polymerase chain reaction method is used to recognise polymorphous variants of IL6 and TGFb1 genes. Recognising the homozygous genotype CC in -174 position of IL6 gene in males and females, as well as the heterozygous genotype GC in -915 position of TGFb1 gene in females enables predicting the high risk of the complicated clinical course of the urogenital Chlamydial infection.
EFFECT: invention enables deciding on reasonable grounds on selecting a therapeutic approach to a specific patient suffering from urogenital Chlamydial infection in order to prevent complications of the urogenital Chlamydial infection and reproductive dysfunctions.
4 tbl, 5 ex
SUBSTANCE: inventions can be used in chemical and jewellery industry. Nitrogen-doped diamond material, obtained in accordance with CVD technology, or representing monocrystal or precious stone, demonstrates difference of absorptive characteristics after exposure to radiation with energy of at least 5.5 eV, in particular UV radiation, and thermal processing at temperature 798 K. Defects into diamond material are introduced by its irradiation by electrons, neutrons or gamma-photons. After irradiation, difference in absorptive characteristics decreases.
EFFECT: irradiated diamond material has absorption coefficient lower than 0,01 cm-1 at 570 nm and is capable of changing its colour.
18 cl, 7 dwg, 11 tbl, 15 ex
SUBSTANCE: ions of carbon with opposite charges interact with each other for 20-30 hours at a temperature of 850-950 °C in a high frequency electro-field in the range of frequencies of 40-80 kHz in the presence of iron as a catalyst. The process is carried out in a melt of salts, containing, wt %: SiC - 7.5-11.0; Na2CO3 or K2CO3 - 89.0-92.5. Applied is granulated iron, which has a size of granules 1-3 mm, in a quantity of 5-10% from the melt weight.
EFFECT: invention makes it possible to simplify the process of the diamond synthesis and its instrumentation, eliminate harmful and dangerous conditions.
1 tbl, 1 ex
SUBSTANCE: initial composition, consisting of the following components, wt %, is prepared: C-60 or C-70 fullerenes - 30-50; heat-conducting component - 10-60; binding agent - the remaining part. The heat-conducting component is selected from the group: wurtzide boron nitride, cubic boron nitride, diamond or their mixtures. The binding agent is selected from elements of the group IVa of the Periodic system or their alloy with copper. The heat-conducting element can be preliminarily covered with the binding agent. The obtained composition is subjected to impact of static pressure from 8 to 13 GPa with heating to 900-2000°C for not less than 20 seconds. A superhard composite material with heat-conductivity to 330 W/m·K, a ratio of microhardness to an elasticity coefficient 0.12 is obtained.
EFFECT: high wear resistance of the material.
2 cl, 3 ex
SUBSTANCE: invention can be used in the field of elaboration of diamond-based materials for magnetic therapy, quantum optics and medicine. A method of determining an angle of misorientation of diamond crystallites in a diamond composite includes placement of the diamond composite into a resonator of an electronic paramagnetic resonance (EPR) spectrometer, measurement of EPR spectrums of nitrogen-vacancy NV-defect in the diamond composite with different orientations of the diamond composite relative to the external magnetic field, comparison of the obtained dependences of EPR lines with the calculated positions of EPR lines of NV-defect in the diamond monocrystal in the magnetic field, determined by the calculation. After that, the angle of misorientation of the diamond crystallites is determined by an increase of width of EPR line in the diamond composite in comparison with the width of EPR line in the diamond monocrystal.
EFFECT: invention ensures higher accuracy of measurements.
3 cl, 6 dwg
FIELD: process engineering.
SUBSTANCE: invention relates to PCD diamond to be used in production of water-jet ejectors, engraving cutters for intaglio, scribers, diamond cutters and scribing rollers. PCD diamond is produced by conversion and sintering of carbon material of graphite-like laminar structure at superhigh pressure of up to 12-25 GPa and 1800-2600°C without addition of sintering additive of catalyst. Note here that sintered diamond grains that make this PCD diamond feature size over 50 nm and less than 2500 nm and purity of 99% or higher. Diamond features grain diameter D90 making (grain mean size plus grain mean size × 0.9) or less and hardness of 100 GPa or higher.
EFFECT: diamond features laminar or fine-layer structure, ruled out uneven wear, decreased abrasion.
15 cl, 5 tbl, 5 ex
SUBSTANCE: invention may be used in medicine in producing preparations for a postoperative supporting therapy. What is involved is the high-temperature decomposition of methane on silicone or nickel substrate under pressure of 10-30 tor and a temperature of 1050-1150°C. The heating is conducted by passing the electric current through a carbon foil, cloth, felt or a structural graphite plate whereon the substrates are arranged. An analogous plate whereon a displacement potential from an external source is sent is placed above the specified plate. Nanodiamonds of 4 nm to 10 nm in size are deposited on the substrates.
EFFECT: higher effectiveness of the method.
1 dwg, 6 ex
FIELD: process engineering.
SUBSTANCE: invention relates to blast processes of the synthesis of materials, in particular, diamonds. Proposed device comprises flow vessel 1 with tight cover 3, mix of explosive arranged inside said vessel that features a high specific energy and graphite or carbon-bearing explosive with negative oxygen balance, initiator 5, indestructible cylindrical barrier 6 composed by pipe arranged aligned with vessel 1 there inside. Note here that said mix of graphite and explosive and initiator 5 are placed at barrier 6 centre.
EFFECT: protection of device wall against maximum loads, increased bulk of explosive without increase in device volume and weight.
FIELD: medicine, pharmaceutics.
SUBSTANCE: present invention refers to pharmacology, nanomaterials and nanotechnology, and concerns a method for selective final purification of nanodiamonds to remove foreign nitrate ions and sulphur compounds to be used in pharmaceutics; the method implies that charge-free nanodiamond powder is treated with alkaline water of the concentration of 0.01-1 mole/l at 20-100°C; the prepared suspension is then decanted and centrifuged; the precipitation is washed with water using ultrasound, separated and dried.
EFFECT: higher effectiveness of the method.
5 cl, 1 dwg, 7 ex
FIELD: process engineering.
SUBSTANCE: invention relates to diamond grinding in making diamond rock cutting tool. Proposed method comprises processing the diamonds in velocity layer of magnetic fields together with ferromagnetic particles. Mix composed of ferromagnetic particles and diamond grains fills the cylindrical case by 0.25-0.35 of its volume. Diamond magnetic susceptibility is defined by the relationship:
EFFECT: higher efficiency of grinding and quality of finished diamonds.
1 cl, 2 tbl, 1 ex
FIELD: process engineering.
SUBSTANCE: invention relates to production of carbon-based superhard composite to be used for making tools for mining, stone-working and metal working. Proposed method comprises applying high pressure and temperature to initial carbon component, a diamond, and binder. Note here that said carbon component comprises additionally fullerene and/or nanodiamond while said binder represents one or several components selected from the family including silicon bronze alloy, Monel metal, solid alloy. Superhard material is produced in two steps. At first step, the mix of initial components is subjected to dynamic pressure of 10-50 GPa at 900-2000°C. At second step, obtained material is placed in high-pressure vessel and subjected to static pressure of 5-15 GPa and heated to 700-1700°C for at least 20 seconds.
EFFECT: superhard micro hardness, high modulus of elasticity and higher wear resistance.
4 cl, 1 tbl, 2 ex
SUBSTANCE: invention relates to colloidal chemistry and can be used in luminescent labels, as well as in production of materials for lasers, light diodes, solar panels, and photocatalysts. First, sodium sulfide and silver nitrate are prepared separately. For this purpose 0.01-0.5 g of sodium sulfide and 0.01-0.5 silver nitrate are dissolved in 40-200 ml of cold distilled water. 0.5-20 g of gelatin swell in reactor for 30 min in 100-500 ml of distilled water with temperature from 20-30°C. Obtained gelatin solution is heated to 40-90°C with mixing, 5 ml of 96% ethanol are poured into it. After that, double-stream pouring of prepared solutions of sodium sulfide and silver nitrate is realised, with further heating for 10-20 min with obtaining sol of colloidal silver sulfide quantum dots and cooling to 4-10°C for 10 hours. Obtained jelly is crushed to size of granules 5-10 mm, washed with distilled water at temperature 7-13°C, excess of water is decanted and granules are heated to temperature higher than 40°C.
EFFECT: invention makes it possible to obtain silver sulfide quantum dots with size 1-5 nm in gelatin matrix, luminescent in the range 800-1100 nm.
2 cl, 4 dwg, 2 ex