Method for measuring maximum allowable blood concentration of heavy metals in children after integrated exposure

FIELD: medicine.

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

 

The invention relates to the field of medicine and is intended to substantiate the maximum allowable concentrations (MAC) of heavy metals in the blood of children living in conditions of polluted habitats, according to the criteria of risk to health in chronic mnogozernovoy exposure. The obtained results can be used in order to increase the effectiveness of planning control and surveillance activities for the objects of the external environment and the management decision-making on provision of sanitary-epidemiological security of the population and protection of the rights of consumers.

There is a method of determining the maximum permissible concentration of pollutants in water bodies (RF Patent No. 2480747), which includes the analysis of water samples taken at sites located in areas with proven environmental well-being, the selective determination of the average concentration of pollutants, determination of maximum permissible concentrations of contaminants carried out taking into account a minimum of contamination by the formula:

where Cn- maximum allowable concentration of pollutants;

Cf- background concentrations of chemicals in waterways;

With awith af - the average concentration of the substance;

SSF- standard deviation of concentration;

tst- ratio t-test at P=0.95;

n is the number of samples taken at the site.

Specified the known method provides a more accurate definition.

However, this method is not intended to define the MPC in the child's blood.

Also, the number of known methods for determining the MPC of heavy metals in the air, namely, the Method of determining the maximum permissible concentration of compounds of metals in the atmospheric air of populated areas (Ed. St. USSR №1660681) and the Method of determining the maximum permissible concentration of aerosols of vanadium pentoxide in the air (Ed. St. The USSR №1140789).

According to the first method the experiment is carried out on two groups of pregnant rats, one of which serves as a control and the other exposed to a toxic agent in a concentration which is much higher estimated safe level of exposure, determine the biochemical parameters in the serum of albino rats, expect steam entropy averages of the indicators in the experimental and control groups, find the average value of the pair entropy metrics and determine MPC mathematical expression.

According to the second method are inhalants effects on laboratory animals and investigate biochemical parameters in biology is Eskom material, moreover, the animal was exposed 3-5 concentrations of vanadium pentoxide, additionally determine in cadaveric organs simultaneously on all groups of animals lipids, proteins and enzymes and on the magnitude of changes define the MAC.

However, the drawback of both these methods is the inability of their application to determine the MAC of heavy metals in the blood of children, because it provides the effect on the body known to be a high concentration of such metal that can cause irreparable harm to the health of the child.

In the prior art have not been identified sources of information about inventions, fully relevant to the claimed technical object, there is no prototype.

The technical result achieved by the invention, is to provide the possibility of determining the maximum permissible concentrations of heavy metals in the blood of children in mnogozernovoy exposure using gentle methods, eliminating harm to the health of the child.

This technical result is achieved by the proposed method of determining the maximum permissible concentrations of heavy metals in the blood of children in mnogozernovoy exposure, according to which choose environmentally disadvantaged area with high load of heavy metals in the environment with the criterion of the hazard index d is I the health of the population is greater than 1; with the specified area produce a representative sample of children for research - a core group, with the use of biological, social and hygienic criteria; using the same criteria produce a representative sample of children in a control group of prosperous in the environmental plan of the territory in these areas using an annual exposition carry out a quantitative assessment of chronic exposure to the studied heavy metal to establish the average daily concentration in environmental objects, and then using these values of average daily concentration of heavy metal in the external environment of hope for the children of both groups on average annual exposure total average daily dose of heavy metal, coming from various sources in the child's body, defining at the same time while leading the way revenues and priority environment, then the children once in three months for one year to produce a sample of blood to establish the following parameters: for the determination of the studied heavy metals and to determine the level of biochemical parameters of blood plasma and blood serum, which characterize the response in the form of actual or potential violations of the health markers is than, under the influence of the heavy metal from the priority environment impact; then obtained from the four results during the year, calculate the average concentrations of the studied heavy metals in the blood and compare it with the reference level for this heavy metal, using a two-sample student test with significance level of 0.05, while setting the adequacy of the choice of the children in the study and control groups; however, the specified criterion of adequacy for the control group of children is the lack of significant differences in the average concentrations of the studied metal in the blood with a reference level, and for the main group of children - the presence of significant differences in the average concentrations of the studied metal in the blood, with an average concentration of this metal in the blood of children in the control group; then use the method of mathematical modeling for children in each group establish the relationship between exposure is the total average daily dose of the investigated metal entering the body from the external environment, and a marker of exposure - the average concentration of heavy metal in the blood, determining the presence of a reliable connection with the exposition of the validity of the selected marker of exposure; then, using the technology of "sliding window" carry out the justification of the selected markers is than through the establishment and evaluation according to the probability of previously established biochemical parameters of plasma and blood serum in children of the main group relative to similar indicators in children control group, and from physiological norms; next, a method based on the analysis of odds ratios, determine the maximum allowable concentration of a marker of exposure and corresponding token response based on the conditions under which the ratio of the odds that characterize the strength of the link between exposure to heavy metal and the response body will be greater than or equal to one for each observation form data table: average concentration of metal in the blood and the values of the markers of response for each observation and produce the conventional division of the data into two parts: below the current observation is the average concentration of heavy metal in the blood and above the current observation; next to both parts of the count value, characterizing the probability of the token is rejected response of the main group from the marker of response in the control group, as the ratio of the number of observations that differ from the control, the total number of observations, and the odds ratio OR for each daily average concentrations of the studied metal in the blood was determined from the relationship:

where i is the index reflecting the number of observations;

pi-- the probability of the token is rejected response core group is s from the marker of response in the control group in the table in the area below the current observation;

pi+- the probability of the token is rejected response of the main group from the marker of response in the control group in the table in the field above the current observation;

moreover, the reliability of the calculated indicator of the odds ratio estimate 95% confidence interval, within which is the true value of the odds ratio, and the relationship recognize reliably set at the lower limit of the confidence interval is greater than or equal to 1; then build a model of dependence between the level of a marker of exposure and the specified ratio of chances, and model building exercise based on the average concentrations of the studied metal in the blood and the data on the value of the odds ratio deviation marker of response in the primary group relative to the value of the marker of response in the control group for each observation; and to establish the adequacy of the model, and therefore reliability of the obtained data, use the procedure dispersion analysis based on the calculation of the Fisher criterion and the coefficient of determination, taking into account that differences considered statistically significant when p≤0,05; next, calculate the maximum reference inoperative the concentrations of heavy metal in the blood, x 0for each token response by the formula:

wherea1,a0the parameters of the model

they also provide a definition when building the model 95-percentage confidence limits point estimates of the fixed concentration of metal in the blood, and as reference concentrations of the studied heavy metals in the blood is the upper 95% confidence limits for each token; and as the maximum permissible concentrations of the studied heavy metals in the blood of children receiving the lowest concentration of the available range of 95%-s upper confidence limits of the fixed metal concentrations in the blood for each token response.

The technical result is achieved due to the following.

For an unambiguous understanding of the used terms and definitions of the present invention are given in the following point:

Marker of exposure is an exogenous substance or its metabolite, the amount of which is determined in biological environments of the body.

The token response is an indicator of quantitatively characterize the biochemical, physiological, behavioral, or other change in the organism, the severity of which is determined by the actual or potential violation of the health or development of the disease./p>

The reference level of daily exposure to chemicals throughout their life, which is installed with all available current scientific evidence and probably does not result in unacceptable health risks to sensitive groups of the population.

External human environment is a set of objects, phenomena and environmental factors, which determines the conditions of human life.

Daily dose/concentration - potential daily dose/concentration, averaged for the whole period of exposure to chemical substances (Sobranie annual exposure).

Exposure time (exposure) - contact body (receptor) with chemical, physical or biological agent.

Maximum permissible concentration (MPC) of heavy metals in the blood of children - maximum allowable concentration of heavy metals per unit volume of blood children, due to mnogozernovoy chronic exposure, which is the daily exposure for indefinitely long time does not cause significant (with a probability of p≤0,05) changes in responses of the body that are installed on the criteria of risk to health.

Due to the fact that the implementation of the proposed method selects the region with ecological problems with high load chemical fact the ditch habitat with the criterion of the hazard index HI for the health of the population is greater than 1 (criterion index HI danger to the health of the population is determined by the "risk assessment Guidance for public health when exposed to chemicals polluting the environment" P 2.1.10.1920-04), provides the definition of residential areas, which in children population in the developing human health from exposure to heavy metals is the most likely.

Use when selecting children in the main and control group multiple criteria: biological, hygienic, social, allows you to generate the most representative sample, the main factor is the difference of which is the presence of mnogozernovoy exposure of heavy metals.

The implementation of a quantitative assessment of chronic exposure to the studied heavy metal, which set quantitative flow of the metal in the organism of children of different ways: oral, inhalation, transdermal, by contact with various environmental objects (water, air, soil, food), it is necessary to correctly calculate the average daily dose of the investigated metal.

Calculating the total average daily dose averaged on an annual exposition, in various ways receipts: inhalation, oral, drinking water, food, studied heavy metals in the body using standard values of exposure factors and specific mass values is eating and age of children included in the main and control group allows you to set the level of impact mnogozernovoy exposure of the investigated metal under chronic exposure with respect to time of exposure, individual characteristics of children and priority entry paths, which increases the information content of the proposed method. The specified calculation is performed according to the formulas presented in Appendix 3 guidelines for the assessment of risk to human health when exposed to chemicals that pollute the environment (R. 2.1.10.1920-04).

The comparative analysis of average daily doses of the investigated metal for the children in the study and control groups, establishing the contribution of each route of entry of the studied metal in the total average daily dose, it is necessary to determine the host path receipts and priority environmental exposure, to determine the affected organs and systems at each intake path in the complex are received simultaneously in several ways. This ensures almost complete priority chemical risk factors for health problems of the child, taking into account their negative impact and increase the reliability of definition.

Due to the quality of the investigated specimens of venous blood is easy and nainstalovani, and obtain the necessary information. The establishment of contents chemical contaminant - heavy metal it is in the blood because the blood is the most homeostatic environment (controllability and adjustability of the concentrations of its constituent components) and only having reviewed constants (reference level) in relation to man-made chemicals. And execution of sampling blood samples in children of the main and control groups at 1 year with a periodicity of 1 times in 3 months and determination of the content of the investigated metal allows us to quantitatively characterize and calculate the average concentrations of the studied heavy metals in the blood, which makes the method more accurate and reliable.

The comparison of obtained values of average daily concentration of the studied metal in the blood sample for each observation (each child in the sample) in the experimental and control group with the reference level (source of information about the values of the reference level is the "risk assessment Guidance for public health when exposed to chemicals, polluting the environment" R. 2.1.10.1920-04), using a two-sample student test (t) with significance level of 0.05, provides for the establishment of the adequacy of the choice of the children in the study and control groups. When atomoxatine criterion of adequacy for the control group of children is the lack of significant differences in the average concentrations of the studied metal in the blood with a reference level, and for the main group of children - the presence of significant differences in the average concentrations of the studied metal in the blood, with an average concentration of this metal in the blood of children in the control group,

Communication between exposure - the total average daily dose of the investigated metal from external factors and the average concentration of heavy metal in the blood, by the method of mathematical modeling, due to the need of calculating the marker of exposure, indicating the presence of a reliable relation witnessed the average concentration of metal in the blood with an exposure that increases the reliability of this method.

Rationale markers of response is necessary in order to determine the extent of the negative impact of the investigated metal to critical organs and systems. And execution of this study by establishing and evaluating the dependence of the probability of these investigational laboratory (biochemical) performance responses in children of the main group compared to the same indicator in children of the control group from the average concentrations of the studied metal in the blood (marker of exposure) using the technology of "sliding window" simplifies the way. To do this, for each value of the concentration of the studied metal in the blood (xi) calculates the probability of QCD is onine laboratory (biochemical) metric from the values in the control (p icalculated to range ("sliding window"):

xi-δ<x≤xi+δ,

where δ is the width of the moving window, which is determined from the relationship:

where N is the total number of studies for the whole population;

xmaxthe maximum concentrations of the studied metal in the blood;

xmin- the minimum concentration of the studied metal in the blood.

Estimation of probability pideviations laboratory (biochemical) index of the core group from the control and from physiological norms produced by the classical formula of probability:

where mia number of studies on the i-th laboratory indicator, deviating from the values in the control range of the xi-δ<x≤xi+δ;

ni- the total number of studies for the i-th laboratory indicator for the range xi-δ<x≤xi+δ.

Graphical illustration of the process of evaluating the probability laboratory (biochemical) indicator from physiological norms using sliding window" is presented in figure 1.

Estimation of parameters according to the probability of laboratory parameter, in relation to the physiological norm, from a daily average concentrations of heavy metal in the blood is by building a logistic regression model:

where p is the probability of deviations of laboratory indicator from the physiological norm;

x - the concentration of a chemical in blood, mg/DM3;

b0b1the parameters of the mathematical model.

Due to the fact that in the blood of every child from both groups define a set of laboratory (biochemical) indicators of specific and nonspecific response of the body, the risk of disease critical organs and systems when possible ways receipt of the studied metal in the body, substantiates the negative impact of the investigated metal on critical organs.

Determination of maximum allowable concentrations of a marker of exposure is based on the analysis procedures of calculation of odds ratios, based on the conditions under which the ratio of the odds OR characterizing the strength of connections between exposure to chemicals and the body's response will be greater than or equal to one. For calculation of odds ratios for each observation form the first data table: average concentration of metal in the blood and the values of the markers of response for each observation (1...n), and in the specified data table to produce the conventional division of the data into two parts: below the current observation - level average concentric and studied metal in the blood and higher current observations - the average concentrations of the studied metal in the blood, for both parts count value characterizing the probability of rejection of the marker response values from similar laboratory indicator in the control group (pi-andpi+respectively) as the ratio of the number of observations that differ from the control, the total number of observations.

And the odds ratios for each average concentrations of the studied metal in the blood was determined from the relationship:

where i is the index reflecting the number of observations.

Moreover, the reliability of the calculated indicator of the odds ratio estimate 95% confidence interval, within which is the true value of the odds ratio, and the relationship recognize reliably set at the lower limit of the confidence interval is greater than or equal to 1.

Due to the fact that next a model of the dependence between the level of a marker of exposure and the specified ratio of chances, and model building exercise based on the average concentrations of the studied metal in the blood and data about the corresponding value and the ratio of the odds of a deviation of the marker response values of the indicator in the control group for each observation, ensures the reliability of the received communication.

Moreover, to establish the adequacy of the proposed model, and therefore the reliability of data obtained using the procedure of dispersion analysis based on the calculation of the Fisher test (F) and determination coefficient (R2taking into account that differences considered statistically significant at p≤0,05.

Next, calculate the reference maximum inactive concentrations of the studied metal in the blood (x0for each token response by the formula:

wherea1,a0the parameters of the model

they also provide a definition when building the model 95-percentage confidence limits point estimates of the fixed concentration of metal in the blood, and as reference concentrations of the studied metal in the blood is the upper 95% confidence limits for each token response, and as a maximum allowable concentration of the studied metal in the blood of children receiving the lowest concentration of the available range of 95%-s upper confidence limits of the fixed metal concentrations in the blood for each token response.

Thus, the achievement of the technical result is achieved by a combination of all features offered from the Britania and excluding any will make it impossible to implement the purpose of the method.

The proposed method is as follows, in the specific example:

1) Choose environmentally disadvantaged areas based on high airborne load of heavy metals. As this site was selected, the Chusovaya river, characterized by the presence of the priority components of emissions of industrial enterprises compounds of manganese in ambient air of residential areas. According to the results of previously conducted risk-assessment procedures for the health of the population in a given area exposure of metals in airborne exposure creates an unacceptable risk of the respiratory system, Central and autonomic nervous system (the index of health risk HI=8-44 at an acceptable level of ≤1,0).

2) Form a comparable sample of children on the basis of biological criteria: I-II health group, which is established on the basis of the results of the analysis of medical cards (form 26) and clinical examination; physiological course of pregnancy and childbirth from the mother; the absence of pathology perinatal period; no burdened hereditary history; vasoreactive indicators Quetelet index, is not beyond ±15%; no acute infectious diseases not less than 3 weeks before the start of the study; the infectivity index of 0.2-0.5; based on the and social criteria: income, family life, quality of living conditions; the parents of the children included in the sample must have secondary and higher education that determines the family life as comparable, the average level of material wealth, housing conditions, meet hygienic standards; on the basis of hygienic criteria: samples are characterized by the exposure of manganese in ambient air of the territory of the children of the sample studied and the lack in the areas of residence sample of the control group; in the presence of common compounds territory comparable. The study group was selected 80 children from organized team, living on the territory, the Chusovaya river. As control group we selected 50 children from organized team, living on the territory of the village Ilyinsky.

3) In the selected areas carry out a quantitative assessment of chronic exposure to the studied heavy metal with the use of annual exposure for the establishment of the average daily concentration in ambient air and drinking water based materials monitoring according to field studies. The average daily concentration of manganese in ambient air ecologically unfavorable territory was 0,0011±0.0003 mg/m31.1 share MAC), on-site comparison - 0,00003±0,00001 mg/m3(0.03 share of MPC). The average daily concentration of manganese in drinking water environmentally damaged site was 0.02±0.003 mg/m3(0.2 share MAC), on-site comparison of 0.01±0.002 mg/m3(0.1 share MAC).

4) using the formulae given in Appendix 3 guidelines for the assessment of risk to human health when exposed to chemicals that pollute the environment (R. 2.1.10.1920-04), calculates the total average daily dose of heavy metal coming from different sources in the body of the child population. Ecological problems of the territory of the average daily intake of manganese in the intake atmospheric air amounted to 9.0·10-4mg/(kg·day), drinking water - 1,1·10-4mg/(kg·day). The total average daily dose was 0.001 mg/(kg·day), and priority admission is atmospheric air. On the territory of comparison, the average daily intake of manganese in the flow of atmospheric air was 2.9·10-5mg/(kg·day), drinking water - 5,5·10-5mg/(kg·day). The total average daily intake of 8.4·10-5mg/(kg·day).

5) children of the specified groups shall conduct sampling of venous whole blood in one tube 1 every 3 months for 1 year (i.e. four times per year) to determine stereoimages in whole blood. When making the last selection of blood samples using three tubes: the first is to determine the metal content, the second is to determine the level of biochemical parameters of blood serum, and the third to determine the level of biochemical parameters of blood plasma, the list of which is determined taking into account the identified affected organs and systems, characteristic of the negative impact of the studied heavy metal manganese and priority of its receipt.

6) In whole blood to determine the level of manganese content on the atomic absorption spectrophotometer Perkin Elmer 3110 using as oxidant acetylene-air mixture with detection mode flame atomization.

In serum with regard to critical organs during chronic inhalation of manganese determine the level of lipid hydroperoxides, the level of immunoglobulin E (IgE) overall, IgE specific for manganese, the level of activity of cytoplasmic superoxide dismutase Cu/Zn-SOD), immunoglobulin G (IgG), A (IgA), cyclic amp C-AMP, cyclic guanidinoacetate C-GMP. In plasma determine the level of malondialdehyde (MDA), an indicator of antioxidant activity (AOA).

7) Calculate an average concentration of manganese in the blood of children during the year (i.e., the amount h of the four indicators, received over the years, divided into four). The data obtained is compared with a reference level. The results obtained are presented in table 1.

The data in table 1 indicate the validity of the sample of children in the study and control groups, because the control group no significant differences in the concentration of manganese in the blood with a reference level (p=0,065, i.e. more than 0.05), and in the main group compared with the control (p=0.001, i.e. less than 0.05.

8) Next, establish a reliable relationship between exposure is the total average daily dose of the investigated metal entering the body from the external environment, and a marker of exposure - the average concentration of heavy metal in the blood, determining the presence of a reliable connection with the exposition of the validity of the selected marker of exposure, method of mathematical modeling. Identification and estimation of the parameters of this dependence allowed us to obtain an adequate model (F≥3,96, p≤0,05) dependence between the average manganese concentration in the blood of the total average daily dose of inhaled and ingested into the body (in the range of investigated concentrations for the same period of observations) (F=2055,12, R2=0.64, p=0,0001). The dependence of the average manganese concentration in blood of children from the total average daily dose of manganese in chronic the integral exposure on-site placement of metallurgical production is presented in figure 2. Based on our data, the average concentration of manganese in the blood of the children of the main group is accepted as a marker of exposure chronic exposure to manganese.

9) To study markers of response perform laboratory examination of children in the study and control groups (through the determination of biochemical parameters plasma and serum). The results are presented in table 2.

The analysis of the obtained results are shown in table 2, revealed the enhancement of nonspecific sensitivity of the organism (sensitization) in a significant increase of 1.3 times the average level of total IgE in serum (103,2±15,6 IU/cm3) relative to the index in children in the control group (79,4±of 4.38 IU/cm3, p=0.001), in 94% of cases exceeding the limit of the physiological norm. Set the initial activation of metabolic process oxidation level of lipid hydroperoxides in serum (362,11±8.6 µmol/DM3), which is 1.2 times higher than children in the control group (301,7±5,23, p=0.02) and in 93% of cases exceeded the limit of the physiological norm. These deviation parameters characterize the development of negative effects - the initial sensitization and activation of metabolic oxidation. The changes in activity of intracellular regulatory intermediaries to increase C-AMP in the serum (,2±0.24 µmol/DM 3) 1.2 times relative to this indicator in the control group (7,73 µmol/DM3, p=0.005) and lower C-GMP in blood serum (2,8±0.11 µmol/DM3) 1.2 times compared to the index of the control group (3,35±0.14 µmol/DM3p=0.005). This indicates activation of the sympathetic elements of regulation at the molecular level. Registered significant excess of 1.4 times the average level of Cu/Zn SOD (102,4±2,42 ng/cm3in serum relative to the index of the control group (72,00±2,43 ng/cm3). In 21% of cases, the figure exceeded the value of the physiological norm, indicating that activation of the antioxidant element.

10) Perform the assay procedure calculation of odds ratios, the indicator of the strength of connections marker of exposure is a marker of response adverse effects". The results are presented in table 3.

11) Build a model of dependence between the level of a marker of exposure and set the ratio of the odds of the procedure of calculation of the maximum inactive average concentration of metal in the blood for each token response. The results are presented in table 4.

From the obtained number of permissible concentrations of manganese in blood for every possible negative response the lowest concentration (lower 95% confidence limit) is 0.01 mg/DM 3(the rate-limiting rate - the probability of increased total immunoglobulin E in serum). This concentration may be recommended as a daily maximum concentration limit of manganese in the blood of children for conditions mnogorazovogo chronic exposure.

Thus, the proposed method provides accurate determination of the maximum permissible concentrations of heavy metals in the blood of children. Knowledge of this information will improve the efficiency of complex sanitary-hygienic measures on the territories with the placement of the objects of metallurgical and machine-building industries and will allow you to assess the real exposure of the population.

Table 1
The average concentration of manganese in the blood of children during the year
MetalGroupManganese concentration (M±M), mg/DM3Reference level (RL), mg/DM3Share RLThe significance of differences (p≤0,05)
with the control groupwith the reference level
control0,010±0,0020,9-0,065
Manganesemain0,042±0,0060,011±0,002the 3.80,0010,0001

Table 2
Laboratory (biochemical) indicators in children with high content of manganese in blood, p≤0,05
Structural levelIndexThe control groupThe main groupThe significance of differences (p)
The mean value and error of the mean (M±M)The recording frequency of samples with deviation from the physiological norm, %The mean value and error of the mean (M±M)The recording frequency of samples that deviate from the physiologically is some norm, %
belowabovebelowabove
MolecularC-AMP in the serum, pmol/cm39.28 are±0,240,08,07,73±0,736,89,00,01
C-GMP in blood serum, pmol/cm32,82±0,110,00,03,35±0,146,82,30,05
IgE total serum, ME/cm3103,2±15,60,094,079,38±of 4.380,033,50,001
IgE special. the manganese in serum, ME/cm31,61±0,030,014,01,61±0,050,016 >0,05
The antioxidant activity of blood plasma, %37,11±1,034,014,642,02±1,9515,045,0>0,05
Cu/Zn superoxide dismutase in serum, ng/cm3102,36±2,420,021,072,0±2,430,012,30,001
Malonic dialdehyde plasma, umol/cm31,84±0,150,014,02,59±0,1125,00,0>0,05
Gidropress lipids in serum, mmol/DM3362,11±8,610,093,0301,7±5,2319,319,30,02
IgG in the serum, g/DM3of 12.53±0,213,010,22±0,2513,118,5>0,05
IgA in serum, g/DM31,45±0,15,07,01,34±0,0718,519,2>0,05

Table 3
The ratio of the odds that characterize the relationship deviates token response with an average manganese concentration in the blood of children
The indicator marker responseGroupThe average concentration of manganese in blood, mg/DM3Figure odds ratios (OR)95% confidence interval (CI)
The decrease of C-AMP in serumcontrol0,010±0,0020,170.104 g÷0,34
main0,042±0,006 1,431,24÷1,90
The increase of C-GMP in blood serumcontrol0,010±0,0020,100,094÷0,11
main0,042±0,0060,520,494÷0,56
The increase in total IgE in serumcontrol0,010±0,0020,570,554÷0,60
main0,042±0,0062,172,044÷2,31
Increased IgE special. the manganese in serumcontrol0,010±0,0020,270,154÷0,35
main0,042±0,0061,821,564÷2,45
the decrease of antioxidant activity of blood plasmacontrol0,010±0,0020,27main0,042±0,0061,560,934÷1,83
The decrease in Cu/Zn superoxide dismutase in serumcontrol0,010±0,0020,300,284÷0,35
main0,042±0,0060,660,624÷0,69
The increase in malondialdehyde plasmacontrol0,010±0,0020,360,054÷0,40
main0,042±0,0061,451,244÷1,90
The increase of lipid hydroperoxides in serumcontrol0,010±0,0020,820,754÷0,95
main0,042±0,0061,101,054÷1,15

Table 4
The model parameters based odds ratios deviations markers of response from the average concentration of manganese in the blood of the children of the main group (p≤0,05)
The token responseThe direction of change of the indicatorThe model parametersThe Fisher test (F)The determination coefficient (R2)The concentration of manganese in blood, mg/DM3
a0a1reference level (upper 95% confidence boundary model)the lower 95% confidence boundary model
Biochemical and immunological parameters
IgE total serumincrease-0,9257,508956,50,630,0150,017
The gidroperekisi lipids in blood serumincrease -0,157,891789,10,350,0170,021
IgE special. the manganese in serumincrease-0,226,474094,30,63to 0.032being 0.036
C-AMP in serumreduction-1,5141,941437,770,680,0340,038
MDA in plasmaincrease-0,041,033876,40,71being 0.0360,042
Antioxidant activity in the blood plasmareduction-0,37at 7.55652,50,660,0500,051

The method of determining the maximum allowable concentric and heavy metals in the blood of children in mnogozernovoy exposure, characterized in that choose environmentally disadvantaged area with high load of heavy metals in the environment with the criterion of the index of risk to human health greater than 1; a specified area produce a representative sample of children for research - a core group, with the use of biological, social and hygienic criteria; using the same criteria produce a representative sample of children in a control group of prosperous in the environmental plan of the territory in these areas using an annual exposition carry out a quantitative assessment of chronic exposure to the studied heavy metal to establish the average daily concentration in the external environment, then using these values of average daily concentration of heavy metal in the external environment of hope for the children of both groups on average annual exposure total average daily dose of heavy metal coming from different sources in the body of the child, while at the same time defining the lead path of receipt and priority environment, then the children once in three months for one year to produce a sample of blood to establish the following parameters: for the determination of the studied heavy IU is Alla and to determine the level of biochemical parameters of blood plasma and blood serum, which characterize the response in the form of actual or potential violations of the health - markers of response, under the influence of the heavy metal from the priority environment impact; then obtained from the four results during the year, calculate the average concentrations of the studied heavy metals in the blood and compare it with the reference level for this heavy metal, using a two-sample criterion Student with significance level of 0.05, while setting the adequacy of the choice of the children in the study and control groups; however, the specified criterion of adequacy for the control group of children is the lack of significant differences in the average concentrations of the studied metal in the blood with a reference level, and for the main group of children - the presence of significant differences in the average concentrations of the studied metal in the blood, with an average concentration of this metal in the blood of children in the control group; then use the method of mathematical modeling for children in each group establish the relationship between exposure is the total average daily dose of the investigated metal entering the body from the external environment, and a marker of exposure - the average concentration of heavy metal in the blood, determining the presence of a reliable connection with the exposition of the validity of the selected Mar the EPA exposure; then, using the technology of "sliding window" provide justification for selected markers of response by establishing and evaluating the dependence of the probability of previously established biochemical parameters of plasma and blood serum in children of the main group as compared to the same indicators in children of the control group and from the physiological norm; then use the method based on the analysis of odds ratios, determine the maximum allowable concentration of a marker of exposure and corresponding token response based on the conditions under which the ratio of the odds that characterize the strength of the link between exposure to heavy metal and the response body will be greater than or equal to one, to do this, for each observation form data table: average concentration of metal in the blood and the values of the markers of response for each observation and produce the conventional division of the data into two parts: below the current observation is the average concentration of heavy metal in the blood and above the current observation; next to both parts of the count value, characterizing the probability of the token is rejected response of the main group from the marker of response in the control group, as the ratio of the number of observations that differ from the control, the total number of observations, and the odds ratios OR for the each daily average concentrations of the studied metal in the blood was determined from the relation:
ORi=pi+1-pi+/pi-1-pi-
where i is the index reflecting the number of observations;
pi-- the probability of the token is rejected response of the main group from the marker of response in the control group in the table in the area below the current observation;
pi+- the probability of the token is rejected response of the main group from the marker of response in the control group in the table in the field above the current observation;
moreover, the reliability of the calculated indicator of the odds ratio estimate 95% confidence interval, within which is the true value of the odds ratio, and the relationship recognize reliably set at the lower limit of the confidence interval is greater than or equal to 1; then build a model of dependence between the level of a marker of exposure and the specified ratio of chances, and model building assests which are based on the average concentrations of the studied metal in the blood and the data on the value of the odds ratio deviation marker of response in the primary group relative to the value of the token response in the control group for each observation; and to establish the adequacy of the model, and therefore, reliability of the obtained data, use the procedure dispersion analysis based on the calculation of the Fisher criterion and the coefficient of determination, taking into account that differences considered statistically significant when p≤0,05; then calculate the reference maximum inactive concentrations of heavy metal in the blood, x0for each token response by the formula:
x0=a0a1(mg/DM3
where a1,a0the parameters of the model
they also provide a definition when building the model 95-percentage confidence limits point estimates of the fixed concentration of metal in the blood, and as reference concentrations of the studied heavy metals in the blood is the upper 95% confidence limits for each token; and as the maximum permissible concentrations of the studied heavy metals in the blood of children receiving the lowest concentration of the available range of 95%-s upper confidence limits of the fixed metal concentrations in the blood for each token response.



 

Same patents:

FIELD: medicine.

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.

2 tbl

FIELD: medicine.

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

FIELD: medicine.

SUBSTANCE: invention concerns diagnosing undifferentiated connective tissue dysplasia (UCTD) in females with a personal history of miscarriage. The technique involves determining the fibrinolytic activity in an endometrial biopsy sample. If the value is less than 23.55 mm2, undifferentiated connective tissue dysplasia is diagnosed.

EFFECT: invention provides the high diagnostic accuracy and enables diagnosing UCTD in the females with the personal history of early miscarriage.

2 tbl, 2 ex

FIELD: medicine.

SUBSTANCE: instrument comprises an oral sample collection vessel, a detector able to detect a marker in this sample, an indicator actuated by a detector signal. The above vessel is detachably connected to an oral cavity instrument. The vessel comprises a sample collection element, a sample storage container, and a passage connecting the collection element and the container to supply the sample to the container by capillary action. The indicator is integrated into the container. The declared instrument is used to diagnose oral diseases by collecting the oral sample, detecting one or more markers in this sample and indicating the presence of one of the disease markers.

EFFECT: inventions enables establishing an accurate and fast diagnosis of the oral pathologies accompanying the daily oral care by placing the detector inside the container able to accumulate a required amount of the sample to be diagnosed.

25 cl, 1 dwg

FIELD: medicine.

SUBSTANCE: technique involves the clinical-laboratory examination of a sportsman who completed heavy physical activity 12-16 hours ago. The examination extent is determined taking into account the organs and systems most vulnerable to the physical activity while deriving the prognostically significant criteria of the morphofunctional body state. The examination involves measuring and analyzing the biochemical, haematological, immunological and functional values, as well as vitamin-mineral saturation. And if the above values are stably unchanged, reliably different from the norm, nonspecific changes of the sportsman's organs and systems are diagnosed.

EFFECT: technique provides the early diagnosis of the significant changes of the organs and systems during trainings and competitions that enables taking further timely measures to prevent the further progression of pathological conditions and maintaining thereby occupational performance and achieving stable high sport results.

FIELD: biotechnology.

SUBSTANCE: efficiency of treatment of patients with high grade non-Hodgkin malignant lymphoma is determined by the likelihood of achieving remission, and 5-year total and relapse-free survival. The method comprises the study of polymorphism G13494A 6th intron of gene TP53 of the patient. In case of revealing in the patient of homozygous genotype G/G in a given locus the low efficiency of treatment is predicted, namely, low likelihood of 5-year survival of the patient and low likelihood of absence of relapse. In the case of revealing in the patient of the genotype A/A or G/A in a given locus, the high efficiency of treatment is predicted, namely the high likelihood of remission and 5-year survival of patient.

EFFECT: invention enables to assess the efficiency of treatment of patients with high grade non-Hodgkin malignant lymphoma on the degree of polymorphism G13494A of 6th intron of gene TP53.

5 dwg, 10 tbl, 2 ex

FIELD: medicine.

SUBSTANCE: invention represents a diagnostic technique for the disturbed thrombocyte aggregation accompanying mucoviscidosis in children involving a thrombocyte aggregation test using the Multiplate aggregometer inducers. Trays with a magnetic mixer and electrodes are added with NaCl 400 mcl at 37°C and immediately added with whole blood 400 mcl from a hirudin test tube, incubated in the chamber for two minutes; the tray is added with 30 mcl of an aggregation inducer specified in a group: soluble thrombin receptor - peptid-6, adenosine diphosphate, arachidonic acid. The thrombocyte aggregation rate is displayed on the screen in the form of a curve, and the sub-curve area U is automatically calculated; the sub-curve area U shows the thrombocyte aggregation state as compared to reference values in the group of healthy children; if the threshold area U has appeared to exceed the reference, the thrombocyte hyperaggregation, while the threshold area U being less than the reference, the thrombocyte hypoaggregation is stated.

EFFECT: invention provides the timely diagnosis of microcirculatory disorders accompanying mucoviscidosis.

2 ex, 1 dwg

FIELD: chemistry.

SUBSTANCE: invention relates to the field of microbiology, namely to a method of microorganism characteristic. The essence of the method consists in the following: (a) obtaining a sample to be tested, about which it is known that it contains or can contain microorganisms; (b) layering the tested sample on a density buffer in a container, where the said density buffer possesses a uniform density from approximately 1.025 to approximately 1.120 g/ml; (c) addition of an identifier into the said tested sample and/or into the said density buffer; (d) centrifugation of the said container in order to separate microorganisms from other components of the said tested sample and to form a deposit of microorganisms; (e) spectroscopic analysis of the deposit and/or the said one or more than one identifier with obtaining measurements, which characterise the microorganisms, where the said spectroscopic analysis is carried out when the said deposit is located in the said container; and (f) characteristic of the microorganisms in the deposit on the basis of the obtained measurements and/or the presence or absence of the said identifier or a metabolised form of the said identifier in the deposit, where the said microorganisms are characterised by one or more classification models, selected from the group, consisting of Gram groups, clinical Gram groups, therapeutic and functional groups.

EFFECT: application of the claimed invention makes it possible to increase the accuracy of the microorganism characteristic.

15 cl, 5 dwg, 1 tbl, 4 ex

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely to qualitative differential instant diagnostic technique for benign and malignant periglottis new growths as shown by oral fluid biomarkers. Substance of the method consists in measuring a quantity of matrix metalloproteinase 2 (MMP 2) in patient's oral fluid; the clinical reference is the level of 1.7-2.9 ng/ml; if the MMP 2 content is 14.4-24.3 ng/ml, patient's periglottis papilloma is diagnosed; if the patient's oral fluid MMP 2 content is 4.1-6.8 ng/ml, periglottis cancer is diagnosed. A biomarker for the qualitative differential instant diagnosis of the periglottis new growths is a tissue inhibitor of metalloproteinase 2 (TIMP 2); the clinical reference is a level of 6.44-11.23 ng/ml; if the TIMP 2 content 29.25-48.75 ng/ml, patient's periglottis papilloma is diagnosed; the TIMP 2 content being 57.23-95.03 ng/ml, periglottis cancer is diagnosed.

EFFECT: using the declared technique enables providing more accurate differential diagnosis of the benign and malignant periglottis new growths.

2 cl, 6 ex

FIELD: medicine.

SUBSTANCE: method is implemented by preparing an incubation solution No.1 containing sulphanilic acid 500 mg in 1 M HCl 50 ml, and solution No.2 consisting of NaNO2 125 ml in distilled water 2.5 ml. Each solution is taken in an amount of 1 ml, mixed in a test tube and added with whole blood with a coagulate 200 mcl. The reaction is carried out for 10 min at a room temperature, and a drop of the suspension is used to produce a multilayer smear on a slide, dried and studied by computed cytophotometry.

EFFECT: more accurate determination.

2 dwg

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.

3 ex

FIELD: medicine, infectology, hepatology.

SUBSTANCE: in hepatic bioptate one should detect products of lipid peroxidation (LP), such as: dienic conjugates (DC), activity of antioxidant enzymes, such as: catalase (CAT)and superoxide dismutase (SOD). One should calculate by the following formula: C = DC/(SOD x CAT)x100, where DC - the content of dienic conjugates, SOD - activity of superoxide dismutase, CAT - activity of catalase. At coefficient (C) values being above 65 one should predict high possibility for appearance of cirrhosis, at 46-645 - moderate possibility and at 14-45 -low possibility for appearance of cirrhosis.

EFFECT: higher accuracy of prediction.

3 ex

FIELD: medicine, clinical toxicology.

SUBSTANCE: at patient's hospitalization one should gather the data of clinical and laboratory values: on the type of chemical substance, patient's age, data of clinical survey and laboratory values: body temperature, the presence or absence of dysphonia, oliguria being below 30 ml/h, hemoglobinuria, erythrocytic hemolysis, exotoxic shock, glucose level in blood, fibrinogen and creatinine concentration in blood serum, general bilirubin, prothrombin index (PTI), Ph-plasma, the state of blood clotting system. The state of every sign should be evaluated in points to be then summed up and at exceeding the sum of points being above "+20" one should predict unfavorable result. At the sum of "-13" prediction should be stated upon as favorable and at "-13" up to "+20" - prediction is considered to be doubtful.

EFFECT: higher accuracy of prediction.

2 ex, 3 tbl

FIELD: medicine, juvenile clinical nephrology.

SUBSTANCE: disease duration in case of obstructive pyelonephritis should be detected by two ways: either by detecting the value of NADPH-diaphorase activity, as the marker of nitroxide synthase activity in different renal department and comparing it to established norm, or by detecting clinico-laboratory values, such as: hemoglobin, leukocytes, eosinophils, urea, beta-lipoproteides, lymphocytes, neutrophils, the level of glomerular filtration, that of canalicular reabsorption, urinary specific weight, daily excretion of oxalates, arterial pressure, and estimating their deviation against average statistical values by taking into account a child's age.

EFFECT: higher efficiency of detection.

7 dwg, 1 ex, 6 tbl

FIELD: clinical medicine, pulmonology.

SUBSTANCE: one should carry out complex estimation of interleukin-1β) concentration in blood, saliva, bronchoalveolar liquid. Moreover, one should detect distribution coefficient (DC) for IL-1β as the ratio of IL-1β blood content to IL-1β salivary content. At increased IL-1β blood content by 10 times and more, by 2 times in saliva, unchanged level of bronchoalveolar IL-1β, at DC for IL-1β being above 1.0 one should predict bronchial obstruction. The method enables to conduct diagnostics of the above-mentioned disease at its earlier stages.

EFFECT: higher efficiency of prediction.

2 tbl

FIELD: medicine, diagnostics.

SUBSTANCE: the present innovation deals with genetic trials, with diagnostic field of oncological diseases due to analyzing DNA by altered status of gene methylation that take part in intracellular regulation of division, differentiating, apoptosis and detoxication processes. One should measure the status of methylation in three genes: p16, E-cadherine and GSTP1 in any human biological samples taken out of blood plasma, urine, lymph nodes, tumor tissue, inter-tissue liquid, ascitic liquid, blood cells and buccal epithelium and other; one should analyze DNA in which modified genes of tumor origin or their components are present that contain defective genes, moreover, analysis should be performed due to extracting and purifying DNA out of biological samples followed by bisulfite treatment of this DNA for modifying unprotected cytosine foundations at keeping 5-methyl cytosine being a protected cytosine foundation followed by PCR assay of bisulfite-treated and bisulfite-untreated genes under investigation and at detecting alterations obtained according to electrophoretic result of PCR amplificates, due to detecting the difference in the number and electrophoretic mobility of corresponding fractions at comparing with control methylated and unmethylated samples containing normal and hypermethylated forms of genes one should diagnose oncological diseases. The method provides higher reliability in detecting tumors, detection of remained tumor cells after operation.

EFFECT: higher efficiency of therapy.

1 cl, 3 dwg, 4 ex

FIELD: medicine, gastroenterology.

SUBSTANCE: one should carry out diagnostic studying, moreover, on the 5th -6th d against the onset of exacerbation in case of gastric and duodenal ulcerous disease one should detect the content serotonin, histamine and acetylcholine in blood, then during 2-3 wk one should conduct medicinal therapy to detect serotonin, histamine and acetylcholine level in blood again and at serotonin content being by 2-3 times above the norm, histamine - by 1.15-1.4 times above the norm and acetylcholine - by 20-45% being below the norm one should predict the flow of gastric and duodenal ulcerous disease as a non-scarring ulcer.

EFFECT: higher accuracy of prediction.

3 ex

FIELD: medicine.

SUBSTANCE: method involves taking blood from ulnar vein (systemic blood circulation) and from large vein of the injured extremity proximal with respect to lesion focus (regional blood circulation). Spontaneous NST-test value is determined and difference is calculated in systemic and regional blood circulation as regional-to-systemic difference. The difference value is used for predicting clinical course of pyo-inflammatory disease in extremities.

EFFECT: high accuracy of diagnosis.

4 cl, 2 tbl

FIELD: medicine, gastroenterology.

SUBSTANCE: one should introduce biologically active substance, moreover, in patient's blood serum one should detect the content of acetyl choline and choline esterase activity followed by 2-h-long intragastric pH-metry at loading with biologically active substance as warm 40-45%-honey water solution at 35-40 C, and at increased content of acetyl choline being above 1.0 mM/l, choline esterase being above 0.5 mM/l/30 min and pH level being 6.0-6.9 it is possible to consider apitherapy to be useful for treating ulcerous duodenal disease.

EFFECT: higher efficiency and accuracy of detection.

3 ex

FIELD: medicine, gastroenterology.

SUBSTANCE: it has been suggested a new method to detect pharmacological sensitivity to preparations as acidosuppressors. After the intake of the preparation a patient should undergo fibrogastroduodenoscopy 3 h later, then, through endoscopic catheter one should introduce 0.3%-Congo red solution intragastrically and the test is considered to be positive at keeping red color that indicates good sensitivity to the given preparation, and in case of dark-blue or black color the test is considered to be negative that indicates resistance to this preparation. The suggested innovation widens the number of diagnostic techniques of mentioned indication.

EFFECT: higher efficiency of diagnostics.

2 ex

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