Method of predicting level of arterial pressure in women in late pregnancy

FIELD: medicine.

SUBSTANCE: invention deals with method of predicting level of arterial pressure in women of Russian nationality, born in Central Black Earth region of Russia. Method includes separation of DNA from lymphocytes of peripheral venous blood and analysis of genetic polymorphisms. +46G/A ADRB2 and 4a/4b eNOS by method of polymerase chain reaction Level of systolic arterial pressure in women in late pregnancy is predicted by results of multiple regression equation of the following type: Y1=15,455+2,544x1+9,946x2+0,736x3+4,716x4+0,185x5, where x1 is genetic variant in locus - 4a/4b eNOS, namely 4b4b=1; 4a4b=2; 4a4a=3; x2 is presence of preeclampsia in relatives: yes=0, no=1; x3 is level of systolic arterial pressure before pregnancy, mm Hg; x4 is presence of cardiovascular system pathology: yes=0, no=1; x5 is woman's weight before pregnancy, kg Level of diastolic arterial pressure in women in late pregnancy is predicted, for which purpose multiple regression equation of the following type is used: Y2=14,200+7,768x1-2,877x2+7,500x3+0,414x4+3,668x5, where x1 is genetic variant in locus - 4a4b eNOS, namely 4b4b+4a4b=1, 4a4a=0; x2 is genetic variant in locus +46G/A ADRB2, namely GG+GA=1, AA=0; x3 is presence of preeclapsia in relatives:yes=0, no=1; x4 is systolic arterial pressure before pregnancy, mm Hg; x5 is presence of cardiovascular system pathology: yes=0, no=1.

EFFECT: invention makes it possible to realise early prediction of increase of arterial pressure level in women in late pregnancy, will make it possible to form of women at the stage of pregravidal preparation and at early terms of pregnancy groups of high risk of developing hypertension in late pregnancy, as well as realise required therapeutic-preventive measures aimed at prevention of development of said pregnancy complication in due time.

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The invention relates to the field of medical diagnostics, can be used to predict the level of blood pressure in women in late pregnancy.

Arterial hypertension (AH) is one of the most common forms of pathology. The frequency of hypertensive States in pregnancy ranges from 15 to 20% and in some regions of Russia up to 29% [vikhlyaeva, E. M. Preclinical manifestations of systemic disorders, clinical outcomes and long-term effects of preeclampsia [Text] /E. M. vikhlyaeva //Obstetrics and gynecology. - 2009. - No. 1. - P. 3-6]. Hypertensive disorders in pregnancy is one of the major public health problems worldwide. The clinical significance of elevated blood pressure in pregnancy due to the high frequency of pathological conditions that develop in pregnant women with high blood pressure [Vertkin A. L., Tkacheva O. N., Murashko L. E. et al. hypertension during pregnancy: diagnosis, management tactics and approaches to treatment [Text]//Attending physician. - 2006. - No. 3. - Pp. 22-25.; Gmurman E. probability Theory and mathematical statistics. - M.: Higher school, 2006. - 284 S.; Makarov O. V., Nikolaev N. N., Volkova E. V. hypertension in pregnancy. Only if preeclampsia? [Text] //M.: GEOTAR-Media, 2006. - 176 p.]. The most dangerous are preeclampsia (5,5%), premature atslog�and normally located placenta (5-10%), intrauterine fetal death associated with placental insufficiency (24,5%). Maternal and perinatal mortality due to complications caused by high blood pressure in pregnant women, is according to who from 20 to 40% [Vertkin A. L., Tkacheva O. N., Murashko L. E. et al. hypertension during pregnancy: diagnosis, management tactics and approaches to treatment [Text] //Attending physician. - 2006. - No. 3. - Pp. 22-25.; Gmurman E. probability Theory and mathematical statistics. - M.: Higher school, 2006. - 284 p.].

Blood pressure is regulated by different mechanisms. To hypertension, along with other factors, may result in the violation of his vascular regulation, associated with reduced nitric oxide in the blood.

Early predictors of these disorders is endothelial dysfunction, the most important markers which include levels of nitric oxide and endothelin-1 [vikhlyaeva, E. M. Preclinical manifestations of systemic disorders, clinical outcomes and long-term effects of preeclampsia [Text] /E. M. vikhlyaeva //Obstetrics and gynecology. - 2009. - No. 1. - P. 3-6]. When "normal" endothelial function balance is always shifted towards the maintenance of vasodilation, which provided, inter alia, due to the stable, sufficient expression of endothelial NO-synthase. This enzyme participates in the synthesis of NO and,consequently, in the regulation of vascular tone and blood pressure. Changes in the expression of NO-synthase are associated with several cardiovascular diseases, including arterial hypertension [Ailamazyan Emergency care in extreme conditions in obstetric practice. [Text] //the 3rd ed. SPb. 2002, pp. 426-432.; H. Critchley, A. Poston, J. Walker, Pre-eclampsia, [Text] //RCOG Press, London (2003), PP. 189-207].

The gene for eNOS localized to chromosome 7q35-36. In the exons and introns of the gene for eNOS found several polymorphic sites, among which the most studied are two, namely minisatellite repeat in intron 4 (eNOS 4a/4b polymorphism) and the mutation at position 298 of the protein sequence, leading to the replacement of the glutamic acid residue to aspartic acid (Glu298Asp). In the case of polymorphic marker 4a/4b eNOS observed correlation between genotypes and the level of nitrates and nitrites in the blood, directly associated with the speed of NO production by the endothelium. The carriers of a genotype 4b/4b have the level of nitrate and nitrite in blood is 25% higher than the carriers of genotype 4A/4A. Thus, it is possible to talk about a potential genetic role of genotype 4A/4A as a risk factor for development of atherosclerosis and diseases, resulting in a loss of normal NO production [Endothelial nitric oxide synthase Glu298Asp gene polymorphism, blood pressure and hypertension in a general population sample [Text] /B. Wolff, H. J. Grabe, C. Schlüter [et al.] //J. Hypertens. - 2005. - Vol.23, No. 7. - R. 1361-1366].

Among the f�Ktorov, influence on blood pressure levels, an important role for β2-adrenergic receptors [Widmaier, E. P. Vander's Human Physiology: the mechanisms of body function [Text] /E. P. Widmaier, H. Raff, and K. T. Strang. - 11th ed. - Boston, MA: McGraw-Hill Higher Education, 2008. - xxviii, 770 p.]. When excited by catecholamines β2-adrenergic receptor encoded by the gene ADRB2, is provided "braking" effect, expressed in the expansion of the blood vessels (coronary, skeletal muscle), smooth muscle relaxation, respiratory tract. Some studies have established the correlation of allele+46G c polymorphism elevated levels of blood pressure and risk of hypertension [Beta(2)-adrenergic receptor gene variation and hypertension in subjects with type 2 diabetes [Text] /K. Bengtsson, M. Orho-Melander, O. Melander [et al.] //Hypertension. - 2001. - Vol.37, No. 5. - P. 1303-1308], other authors found no such Association [N. Kato et al., 2001; S. Herrmann et al., 2002]. Some data are available showing that the polymorphism+46G/A ADRB2 reduces the sensitivity of the receptor to the action of ligands and may be associated with the development of arterial hypertension [Brodde, O. E. Beta1 - and beta2-adrenoceptor polymorphisms and cardiovascular diseases [Text] /O. E. Brodde //Fundam. Clin. Pharmacol. - 2008. - Vol.22, No. 2. - P. 107-125].

To assess the current patent situation was searched by the security documents for the period from 1990 to 2013, the Analysis of documents was made on a method for predicting the level of blood pressure on the basis of molecular genetic data coils� from polymorphic markers of genes.

In examined medical research and the available patent literature, the authors found no way of predicting blood pressure in women in late pregnancy on the basis of data on genetic polymorphisms 4A/4b eNOS and+46G/A ADRB2 in combination with other factors that influence the level of blood pressure.

A prototype of the selected RF patent №2473912 at the request of the Russian Federation No. 2011139871/15, 30.09.2011 "a method for predicting the level of blood pressure in pregnant women depending on the genetic polymorphism of endothelin - 1 (Kurnosov M. I., Reshetnikov, E. A., L. Y. Akulova; Kolesnikov Y., Polnikov A.V.). The claimed method includes extraction of DNA from lymphocytes of peripheral venous blood and the analysis of the polymorphism K198N gene endothelin 1 (ET-1) by polymerase chain reaction. In case of detection of allele C polymorphism K198N ET-1 predict the risk of high blood pressure in pregnant women at the time of 37-40 weeks, is providing an effective criterion for evaluating changes in blood pressure in pregnant women, allowing to pregnancy to determine the probability of changes in blood pressure in the direction of its increase and apply certain tactics of women during the entire gestation period.

The disadvantage of this method is that this analysis is only one genetic factor genetic �olymorphism endothelin -1, associated with blood pressure in pregnant women and does not address other predictors predict blood pressure in women in late pregnancy.

The object of the invention is to expand the Arsenal of methods for predicting blood pressure in women in late pregnancy.

The technical result - obtaining of the evaluation criteria to predict blood pressure in women of Russian nationality, which urozhencami Central Chernozem region of Russia, in late pregnancy, allowing, including before pregnancy, to determine the probability of changes in blood pressure in women when a pregnancy occurs in the direction of its increase and therefore implement certain tactics of women at risk during the entire gestation period.

In accordance with the assigned task has been developed a method for predicting the level of blood pressure in women of Russian nationality, which urozhencami Central Chernozem region of Russia in late pregnancy, including:

- extraction of DNA from peripheral venous blood;

analysis of polymorphisms by polymerase chain reaction;

- forecasting the level of blood pressure in women in late pregnancy, depending on the identified genetic�fir variants at loci 4A/4b eNOS and+46G/A ADRB2 and the following predictors:

for the GARDEN - a woman's weight before pregnancy, the presence of PE with relatives, the presence of pathology of the cardiovascular system, level GARDEN to the pregnancy, a genetic variant at the locus 4A/4b eNOS;

for dad - a woman's weight before pregnancy, the presence of PE with relatives, the presence of pathology of the cardiovascular system, level GARDEN to the pregnancy, a genetic variant at the locus 4A/4b eNOS and genetic variant at locus+46G/A ADRB2.

- prediction of systolic blood pressure in women in late pregnancy on the results of the multiple regression equation of the following form:

Y1=15,455+2,544 x1+9,946 x2+0,736 x3+4,716 x4+0,185 x5,where x1genetic variant at the locus - 4A/4b eNOS (4b4b-1; 4a4b-2; 4a4a-3), x2- the presence of preeclampsia in family (Yes-0; no-1), x3- systolic blood pressure before pregnancy, mm Hg.CT., x4- the presence of pathology of the cardiovascular system (Yes-0; no-1), x5- a woman's weight before pregnancy, kg.

- forecasting the level of diastolic blood pressure according to the results of a multiple regression equation of the following form:

Y2=14,200+7,768 x1-2,877 x2+7,500 x3+0,414 x4+3,668 x5,

where x1genetic variant at the locus 44b eNOS ((4b4b+4a4b)-1; AA-0), x2genetic variant at locus+46G/A ADRB2 ((GG+GA)-1 AA-0), x3- the presence of preeclampsia in family (Yes-0; no-1), x4- systolic blood pressure before pregnancy, mm Hg.CT., x5- the presence of pathology of the cardiovascular system (Yes-0; no-1).

Novelty and inventive step lies in the fact that the prior art not known to predict the possibility of blood pressure in women of Russian nationality, which urozhencami Central Chernozem region of Russia in late pregnancy by the presence of genetic variants 4A/4b eNOS and+46G/A ADRB2 in combination with other predictors.

The method is as follows:

DNA extracted from samples of peripheral venous blood of the patient in 2 stages. In the first stage to 4 ml of blood add 25 ml of lyse buffer containing 320 mm sucrose, 1% Triton X-100, 5 mm MgCl2, 10 mm Tris-HCl (pH=7,6). The resulting mixture was stirred and centrifuged at 4 º C, 4000 rpm for 20 minutes. After centrifugation the supernatant decanted, to the residue add 4 ml of a solution containing 25 mm EDTA (pH=8.0) and 75 mm NaCl, resuspension. Then add 0.4 ml 10% SDS, 35 ál proteinase K (10 mg/ml) and incubated the sample at 37º for 16 hours.

In the second phase from the resulting lysate sequentially perform the extraction of DNA equal volumes of phenol, phenol-chloroform (1:1) and chloroform by centrifugation at 4000 rpm in t�within 10 minutes. After each centrifugation produce a selection of the aqueous phase. DNA is precipitated from the solution with two volumes of chilled 96% ethanol. Formed DNA is dissolved in twice-distilled, deionized water and stored at -200S.

The selected DNA is then subjected to polymerase chain reaction using standard oligonucleotide primers (table 1).

The structure of the primers and probes used for genotyping of the studied DNA markers

Table 1

Name5'-3' sequence of the primers
and probes
Literary source
VNTR 4a/4b eNOS (rs699)F: 5-agg ccc tat ggt agt gcc ttt-3
R: 5-tct ctt agt gct gtg gtc ac-3
Spiridonova, M. G., et al., 2002, 2006
+46G/A ADRB2
(rs1042713)
F: 5'-cgg cag cgc ctt ctt g-3'
R: 5'-tgc gtg acg tcg tgg tc-3'
5'- ROX-cac atg cca gaa gcc - BHQ2-3'
5'- FAM-cac cca ata gaa gcc - RTQ1-3'
D. E. Lanfear et al., 2005

The invention is characterized by:

Fig.1. Discrimination of alleles by locus+46G/ADRB2 A method detection TaqMan probes according to the values of the UOF (relative fluorescence) of each probe on the amplifier IQ5 c detection system in real �belts, wherehomozygotes for an allele+46A,homozygotes for an allele+46G,heterozygotes+46GA,- negative control

Fig.2. Electrophoretic separation of the amplification products VNTR polymorphism 4a/4b eNOS., where 3, 5, 6 homozygotes 4b4b; 2 homozygotes 4a4a; 1, 4 heterozygotes 4a4b.

The possibility of using the proposed method to assess the level of blood pressure in women in late pregnancy confirms the analysis of the survey of 452 women: 249 pregnant women who have observed the increasing pressure of the end of the pregnancy, and 203 women in the control group with normal pregnancy. In the studied sample included individuals of Russian nationality, which urozhencami Central black earth region of Russia and have no relationship among themselves. The average age of the core group 27,11±is 6.42 years (ranged from 18 to 44 years), in the control of 26.5±6,36 years (ranged from 18 to 42 years) (p>0,05). Thus, the population control group did not differ from the main group by age, sex, nationality and place of birth. All clinical and clinical-laboratory research was conducted on the basis of the Perinatal center of the Belgorod regional clinical hospital of Saint Joasaph, with informed consent of the patient� on the use of materials and treatment measures undertaken, during the period of hospitalization and after for research purposes and was recorded by the ethical standards Committee of the Russian Federation.

Using the methods of mathematical modeling multiple regression found a significant influence of genetic, biomedical, and other factors on blood pressure in pregnant women (table.2).

The multiple regression coefficients and level of significance of indicators used to predict blood pressure in women in late pregnancy.

Table 2

Independent (explanatory) signs (Xi) and their gradationsThe regression coefficients for the GARDEN, andi(R)Regression coefficients for DBP, andi(R)
Constant15,455 (0,05)14,200 (0,05)
A genetic variant at the locus 4A/4b eNOS
(4b4b-1; 4a4b-2; 4a4a-3)
2,544 (0,04)-
A genetic variant at the locus 4A/4b eNOS
((4b4b+4a4b)-1; AA-0),
-7,768 (0,001)
Genetic vari�HT by locus+46G/A ADRB2
((GG+GA)-1; AA-0),
--2,877 (0,03)
The presence of pre-eclampsia with relatives (Yes-0;no-1)9,946 (0,00000)7,500 (0,00000)
Systolic blood pressure before pregnancy, mm Hg. article0,736 (0,0000)0,414 (0,0000)
The presence of pathology of the cardiovascular system
(Yes-0;no-1)
4,716 (0,01)3,668 (0,003)
A woman's weight before pregnancy, kg0,185 (0,002)-

To statistically significant (p<0,05) factors that influence the level of blood pressure in women in late pregnancy are: for the GARDEN - a woman's weight before pregnancy, the presence of PE with relatives, the presence of pathology of the cardiovascular system, level GARDEN to the pregnancy, a genetic variant at the locus 4A/4b eNOS; for DBP - a woman's weight before pregnancy, the presence of PE with relatives, the presence of pathology of the cardiovascular system, level GARDEN to the pregnancy, a genetic variant at the locus 4A/4b eNOS and genetic variant at locus+46G/A ADRB2. The result of multiple regression analysis is the calculation of estimates �agressivnyh coefficients a 1and2...andiequation [Rebrova O. Yu., 2006]:

Y=C+a1x1+a2x2+a3x3+...+anxnwhere xi- informative signs, the values of aithe coefficients for these signs, is a constant.

Regression model for predicting systolic blood pressure includes the following predictors: a woman's weight before pregnancy (t=3,03; p=0.002), the presence of PE with relatives (t=6,72; p<0,0001), the presence of pathology of the cardiovascular system (t=2,50; p=0.012), level GARDEN before pregnancy (t=10,16; p<0,0001), a genetic variant at the locus 4A/4b eNOS (t=2.02; p=0.04). These factors determine the=40,30% of the variance of systolic blood pressure in women in late pregnancy, F(5,340)=45,90; p<0,0001. It should be noted that when manafactory models, the contribution of these predictors of variability in systolic blood pressure in pregnant women at the time of 37-40 weeks, estimated using simple regression is significantly lower and amounts to h2=10,27% for weight women before pregnancy (t=6,29; p<0,0001), h2=14,02% for the presence of PE with relatives(t=7,52, p<0,0001), h2=26,61% for systolic blood pressure before pregnancy (t=11,22; p<0,0001).

The multiple regression equation for predicting systolic blood pressure in women in late pregnancy they�et following form:

Y1=15,455+2,544 x1+9,946 x2+0,736 x3+4,716 x4+0,185 x5,

where x1genetic variant at the locus - 4A/4b eNOS (4b4b-1; 4a4b-2; 4a4a-3), x2- the presence of preeclampsia in family (Yes-0; no-1), x3- systolic blood pressure before pregnancy, mm Hg.CT., x4- the presence of pathology of the cardiovascular system (Yes-0; no-1), x5- a woman's weight before pregnancy, kg.

The obtained regression model predicting diastolic blood pressure includes five predictors: the systolic blood pressure before pregnancy (t=8,63; (p < 0,00001), the presence of PE with relatives (t=7,47; p < 0,00001), the presence of the pathology of the cardiovascular system (t=2,94; p=0.003), a genetic variant at the locus 4A/4b eNOS (t=3,31; p=0.001), a genetic variant at locus+46G/A ADRB2 (t=2.06; p=0.03), which determine 34,68% of the variance in the level of diastolic blood pressure in women in late pregnancy (F(5,335)=35,57; p<0,0001. Monofactorial the effects of these predictors account for h2=14,02% for the presence of PE with relatives (t=7,94; p=0,0000), h2=19,79% for systolic blood pressure before pregnancy (t=9,25; p<0,0001).

A multiple regression equation to predict the level of diastolic blood pressure in women in late pregnancy has the following form:

Y2=14,2007,768 x 1-2,877 x2+7,500 x3+0,414 x4+3,668 x5,

where x1genetic variant at the locus 44b eNOS ((4b4b+4a4b)-1; AA-0), x2genetic variant at locus+46G/A ADRB2 ((GG+GA)-1; AA-0), x3- the presence of preeclampsia in family (Yes-0; no-1), x4- systolic blood pressure before pregnancy, mm Hg.CT., x5- the presence of pathology of the cardiovascular system (Yes-0; no-1).

To assess the health of specific regression models we examined the patient in relation to the factors that influence the level of blood pressure.

So, a pregnant E. Russian nationality, which is a native of Central Chernozem region of Russia, the following indicators: the presence of pathology of the cardiovascular system (1), the presence of PE with relatives - (1), GARDEN before pregnancy to 90 mm Hg.article, a woman's weight before pregnancy - 71 kg., a genetic variant at the locus - 4A/4b eNOS (4b4b-1), a genetic variant at locus+46G/A ADRB2 (AA-1). Substituting these characteristic values in the above two equations and find in each equation the value y:

For the GARDEN:

Y1=15,455+2,544*1+9,946*1+0,736*90,0+4,716*1+0,185*71=112,036.

For DBP:

Y2=14,200+7,768*1 - 2,877*0+7,500*1+0,414*90+3,668*1=70,396.

Further monitoring of this patient showed that the level of SBP and DBP at the end of pregnancy was 110,00 mm Hg.PT. and 70,00 mm Hg.PT. soo�respectively.

Therefore, using the proposed method allows to predict the increase in blood pressure in women in late pregnancy.

Thus, the developed models give us the ability to predict elevated levels of blood pressure in women of Russian nationality, which urozhencami Central Chernozem region of Russia in late pregnancy depending on genetic variants loci 4A/4b eNOS and+46G/A ADRB2 in combination with other predictors. Early prediction of increasing blood pressure in women in late pregnancy will enable you to generate among women with pregravid preparation and early pregnancy at high risk of development of hypertension in late pregnancy and deliver in these groups the necessary treatment and preventive measures for the prevention of the development of pregnancy complications.

A method for predicting the level of blood pressure in women of Russian nationality, which urozhencami Central Chernozem region of Russia in late pregnancy, including extraction of DNA from peripheral venous blood, the analysis of polymorphisms by polymerase chain reaction, characterized in that carry out the analysis of genetic polymorphisms +46G/A ADRB2 and 4a/4b eNOS in combination with other pre�cerami and predict systolic and diastolic blood pressure in women in late pregnancy, thus to predict the systolic blood pressure in women in late pregnancy using a multiple regression equation of the following form:
Y1=15,455+2,544 x1+9,946 x2+0,736 x3+4,716 x4+0,185 x5,
where x1genetic variant at the locus - 4A/4b eNOS, namely 4b4b=1; 4a4b=2; 4a4a=3; x2- the presence of preeclampsia in family: Yes=0, no=1; x3- systolic blood pressure before pregnancy, mm Hg.St.; x4- the presence of pathology of the cardiovascular system: Yes=0, no=1; x5- a woman's weight before pregnancy, kg;
- to predict the level of diastolic blood pressure in women in late pregnancy using a multiple regression equation of the following form:
Y2=14,200+7,768 x1-2,877 x2+7,500 x3+0,414 x4+3,668 x5,
where x1genetic variant at the locus 44b eNOS, namely 4b4b+4a4b=1, AA=0; x2genetic variant at locus+46G/A ADRB2, namely GG+GA=1, AA=0; x3- the presence of preeclampsia in family: Yes=0, no=1; x4- systolic blood pressure before pregnancy, mm Hg.St.; x5- the presence of pathology of the cardiovascular system: Yes=0, no=1.



 

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2 ex

FIELD: medicine.

SUBSTANCE: group of inventions relate to medicine and deals with method of diagnosing neurodegenerative disease in individual, including the following stages (i) determination of one or several parameters, selected from group, consisting of 3ab40 or value of calculated parameter, selected from group, consisting of 2ab40+3ab40, 2ab40+3ab40+2ab42+3ab42 and 1ab40+2ab40+1ab42+2ab42; (ii) comparison of parameter value with standard value, corresponding to value of said parameter in standard sample; and (iii) diagnostics of neurodegenerative disease, in case if increase of parameter value in comparison with standard value is observed. Group of inventions also deals with method of detecting stage, preceding neurodegenerative disease, method of differentiating neurodegenerative disease from stage, preceding said neurodegenerative disease.

EFFECT: group of inventions provide high sensitivity and specificity of detection methods.

13 cl, 12 ex, 14 dwg, 12 tbl

FIELD: medicine, ophthalmology.

SUBSTANCE: in lacrimal liquid one should detect the content of interleukin 8 (IL-8) and that of interleukin 1 beta (IL-1β) to calculate prognostic coefficient (PC) due to dividing the first value by the second one by the following formula: At PC value being below 10.0 one should predict favorable disease flow, and at PC value being above 10.0 - unfavorable flow.

EFFECT: higher accuracy of prediction.

2 ex

FIELD: medicine, medicinal microbiology.

SUBSTANCE: method involves growing microorganism culture to be studied in solid nutrient medium followed by preparing microbial suspension and its incubation in the presence of lactoferrin. Control sample is prepared in parallel series. Control and experimental samples are incubated, supernatant is removed from bacterial cells and lactoferrin concentration is determined in supernatant of experimental and control sample by immunoenzyme analysis. Then anti-lactoferrin activity is calculated by difference of concentrations of residual lactoferrin in experimental and control samples. This method provides enhancing the sensitivity and precision in carrying out the quantitative evaluation of anti-lactoferrin activity in broad spectrum of microorganisms that is urgent in diagnosis and prognosis of diseases with bacterial etiology. Invention can be used in determination of persistent indices of microorganisms for assay of their etiological significance in pathological processes.

EFFECT: improved assay method.

3 tbl, 3 ex

FIELD: medicine, biology.

SUBSTANCE: invention relates to nutrient medium used for accumulation of cells for the following cytological and/or immunocytochemical analysis carrying out. Invention relates to medium containing salts NaCl, KCl, anhydrous CaCl2, MgSO4 x 6 H2O, MgCl2 x 6 H2O, Na2HPO4 x 2 H2O, KHPO4, NaHCO3, and also glucose and Henx's solution, 10% albumin solution and polyglucin taken in the ratio 1:1:1. Invention provides enhancing the preservation of cells.

EFFECT: improved an valuable properties of nutrient medium.

3 ex

FIELD: medicine, cardiology.

SUBSTANCE: in peripheral blood one should detect the level of CD95(+) and CD16(+) neutrophilic granulocytes and at combination of increased level of CD95(+) neutrophilic granulocytes by 4 times and more and CD16(+) neutrophilic granulocytes by 0.6 times against the norm with ECG signs of myocardial infarction one should predict lethal result of large-focal myocardial infarction.

EFFECT: higher accuracy of prediction.

FIELD: medicine, parasitology.

SUBSTANCE: one should carry out immunoenzymatic assay to detect diagnostic optic density and that of labeled immune complex in a plot's hole with tested serum measured in conventional units at wave length being 492 nm. One should calculate coefficient of antibodies concentration measured in conventional units by the following formula: CAC = (Odtsh - Odd) x 100, where CAC - coefficient of antibodies concentration, Odtsh - optic density of the hole with tested serum, Odd - diagnostic value of optic density, 100 - coefficient of serumal dilution. By CAC value one should detect the titer of antibodies to Lamblia intestinalis antigens to interpret results of the trial. The method enables to study the dynamics of disease flow.

EFFECT: higher efficiency and accuracy of diagnostics.

1 ex, 1 tbl

FIELD: medicine.

SUBSTANCE: the present innovation deals with studying and treating diseases of inflammatory, autoimmune and degenerative genesis. One should perform sampling of heparinized blood followed by its sedimentation to obtain blood plasma with leukocytes and centrifuging to isolate the latter which are washed against erythrocytic and serumal admixtures, and, also, it deals with calculating the number of cells in samples out of leukocytic suspension after incubation (B) for 1.5 h at 37 C in holes of plastic microplotting board, out of leukocytic suspension one should additionally prepare two samples, one should be applied to calculate total number of leukocytes before incubation (A), the second sample undergoes incubation at the same mode at addition of autoserum to calculate the number of cells remained after incubation (C). One should state upon adhesive properties of leukocytes by the index of spontaneous adhesion (D), where D=(A-B)/B.100%, and effect for enhanced cellular adhesion under the impact of autoserum should be detected by the value of K=(B-C)/C.100% at K ≥ 30%, where B - C - the number of cells undergone additional adhesion after addition of autoserum. The present innovation widens functional possibilities of the suggested method due to obtaining additional values depicting adhesive properties of blood leukocytes.

EFFECT: higher accuracy of detection.

FIELD: medicine, immunology.

SUBSTANCE: one should carry out reaction of blast-transformation, detect proliferation of T-lymphocytes activated with antibodies to CD3 in the presence of interleukin-7 (ACT IL-7) and in the presence of interleukin-7 and dexametazone (ACT IL-7 D), calculate the index for dexametazone action as the ratio of ACT IL-7 to ACT IL-7 D, moreover, the value of dexametazone action index being above 1.2 indicates increased production of cytokins that suppress T-lymphocytes in neonatals. The method enables to detect functional defect of immune system that characterizes neonatal period.

EFFECT: higher efficiency of detection.

2 ex

FIELD: medicine.

SUBSTANCE: method involves measuring forced exhalation volume per 1 s (FEV1) in l, full right ventricle evacuation time (RVE) in ms and angiotensin II value (AII) in ng/l. Discriminant relationship is built as D=0.504·RVE+3.038·FEV1 - 2.0·AII. D being less than 83.88, pulmonary hypertension occurrence is predicted within 1 year. D being equal to or greater than 83.88, no pulmonary hypertension is predicted to occur.

EFFECT: enhanced accuracy of prediction.

FIELD: medicine, medicinal immunology.

SUBSTANCE: method involves determination of heterophilic antibodies in human serum blood by the Paul-Bunnel's method relatively the level of circulating immune complexes, complement-activating properties of heterophilic antibodies by incubation of standardized ram erythrocytes with 0.8% serum for 30 ± 5 min and the following measurement of the erythrocytes lysis degree. The measurement of the effector function coefficient of heterophilic antibodies is carried out by the complement system Keff.f.h.a.-c.s. by the formula: Keff.f.h.a.-c.s. = Y/Tg.a. wherein Y means a lysis degree, %; Tg.a. means a reverse titer of heterophilic antibodies to ram erythrocytes. The damage assay is carried out by comparison of the immune status with the relative level of circulating immune complexes in serum. Method provides detection of preclinic from of immunodeficiency and autoimmune diseases that opens the possibility for their prophylaxis at most early stages of development. Invention can be used for assay of damage in the immune status in human serum blood.

EFFECT: improved method for assay.

5 tbl, 1 ex

FIELD: medicine.

SUBSTANCE: method involves concurrently examining anti-inflammatory IL-4 level in blood serum and lacrimal fluid. The value being within the limits of 60-70 pg/l in blood serum and 5-15 pg/l in lacrimal fluid, disease prognosis is considered to be unfavorable. The IL-4 concentration being within the limits of 90-100 pg/l in blood serum and 20-30 pg/l in lacrimal fluid, disease prognosis is considered to be favorable.

EFFECT: high accuracy of diagnosis.

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