Method for prediction of reduced transport function of band-3 protein of red-cell membranes of newborns delivered by mothers suffered aggravated herpes viral infection in third trimester of pregnancy
SUBSTANCE: invention describes a method for prediction of reduced transport function of band-3 protein of red-cell membranes of newborns delivered by mothers suffered aggravated herpes viral infection in the third trimester of pregnancy wherein newborn's peripheral blood is examined for the TNFa content; band-3 protein is detected in the newborn's red-cell membranes by disk electrophoresis in polyacryl amide gel (10%) with sodium dodecyl sulphate; a discriminant function is calculated by formula Df=(5.167×band-3)+(2.196×TNFα), wherein Df is the discriminant function, band-3 is red-cell membrane stromal protein, TNFα is newborn's blood interleukine, and if Df is more than 218.67, reduced transport anion function by protein band-3 through newborn's red-cell membranes is predicted.
EFFECT: invention provides detecting the effect of herpes viral infection on the content of protein band-3 in newborn's red-cell membranes.
The purpose of the invention is to forecast the sustainability of transfer of anions through the membrane of the red blood cells of newborns during exacerbation of the mother in the third trimester of pregnancy herpes viral infection, increase in the blood content of TNFα, which leads to decrease in the content in the erythrocyte membrane protein band-3.
Protein bands-3 - transport glycoprotein of the erythrocyte membrane (Vorob'ev A.I., 2007 ; Braton D., Cohec S., 1981 ). This is the largest fraction of proteins in erythrocyte membrane was 27% of the total number of proteins in membranes. Protein bands-3 is actively involved in the transport of chloride and bicarbonate across the membrane of erythrocytes (Vorob'ev A.I., 2007 ; Gallagher P.O., Forget C., 2001 ). Protein bands-3 interacts with ancyranum located on the inner surface of the membrane, providing a durable connection of the lipid layer with peripheral proteins (Marchesi V.F., 1979 ; Prankerd T.A., 1961 ; Walensky L., Nariam, Luxs, 2003 ).
The prototype we chose studies Metlenko, Iainglasgow "a Method for predicting the stability of erythrocytes in the peripheral blood during exacerbation of herpes-virus infection in pregnant women" (patent 2344423, registered in the State register of inventions of the Russian Federation on January 20, 2009 ). In this paper, the authors showed that the exacerbation of herpes viral infection with an increase in the membranes is of eritrotsitov the effective absorption with added reduces the protein bands-3.
In this application we draw attention, in contrast to the above studies, that:
1. During exacerbation of herpes virus infection in the mother of the titer of antibodies to the herpes virus builds up in the blood of the newborn;
2. In the blood of the newborn dramatically increases the concentration of interleukin TNFα, which increases the damage of erythrocyte membranes;
3. In previous work, attention was drawn to the erythrocytes pregnant. In the present invention it is noted that this affected the structure and the membrane of the red blood cells of newborns, resulting in reduced transport function of anions using a protein band-3 through erythrocyte membranes newborn.
Studying the available literature, we found no information, highlighting that during exacerbation of herpes viral infection during pregnancy, newborn, under the influence of herpes viruses, activates interleukin TNFα, under whose influence decreases in erythrocyte membranes newborn protein bands-3 that creates a risk of exchange between erythrocytes and blood plasma anions CL-and bicarbonate, which leads to disruption buffer properties in the peripheral blood of newborns.
The claimed method has the following methods.
1. The study was conducted on 20 pregnant women in the third trimester of gestation and 15 newborns, radisys is from these mothers, who in the third trimester suffered exacerbation of herpes virus infection. The titer of antibodies to the herpes virus was determined by ELISA on spectrophotometer "Stat Fax-2100" (USA).
2. The content of TNFα in peripheral blood was determined spectrophotometrically by ELISA on device (Stat Fax-2100" (USA).
3. In erythrocyte membranes newborn method disc electrophoresis in a stepwise gradient polyacrylamide gel (10%) sodium dodecyl sulfate revealed a protein band-3 using densitometry apparatus Biometra (Germany).
4. Research was conducted on the basis of hospital obstetric clinic of the far Eastern scientific center of physiology and pathology of respiration SB RAMS. All studies were conducted with regard to the requirements of the Helsinki Declaration of the world Association "Ethical principles for medical research involving human subjects" with amendments of 2000, and the "Rules of clinical practice in the Russian Federation", approved by Order of Ministry of health of the Russian Federation dated 19.06.2003, No. 266.
Studies have shown that in the case of the exacerbation of herpes viral infection in a pregnant woman in the third trimester of gestation high titers of antibodies in the peripheral blood is celebrated both mother and the newborn (1:12800). Against this background, there has been an increasing activity of interleukin TNFα to 95.05 ng/ml (to the Troll - 22.5±0.85 ng/ml; p<0.001). In the study against this background, the method of disc electrophoresis in polyacrylamide gel (10%) with sodium dodecyl sulfate in erythrocyte membranes newborn found a significant reduction of protein bands 3 to 12.58±0.35% (control - 14.1±0.68%, p<0.001).
Conducted discriminant analysis showed that the discriminant function (DF) is greater than the boundary values during exacerbation of herpes infection and activation of TNFα, influenced the protein content of band 3 in erythrocyte membranes. When DF is more 218.67 predict a decrease of the transport function of anions using a protein band 3 erythrocyte membranes newborn.
DF (273.06)> the boundary values (218.67)
Therefore, if DF (273.06)> the boundary values (218.67) develops on the background of decrease protein bands-3 instability of the transport function of the protein band 3 in erythrocyte membranes.
1. Vorob'ev A.I. Guide to Hematology. M.: Izd. "Novamed". 2007. - 1224 S.
2. Lutsenko M.L., Andrievsky I.A. Patent 2344423, registered in the State register of inventions of the Russian Federation on January 20, 2009
3. Braton D., Cohen C.M., Tjler I. Interaction of cytoskeletetal on the human erythrocyte membrane. Cell. 1981, Vol.24. - P.24-32.
4. Gallagher P.O., Forges B.G. The red cell membrane. Williams Hematology. Ed. E.Beutler, M.A.Littman, B.S.Coller. Chapter 49, 6 editions McGrow Hill International Editon. 2001. - P.333-343.
5. Marchesi V.F. Functional proteins of the human red cell membrane. Semin. Hematol. 1979. - Vol.16. - P.3-20.
6. Prankerd T.A. The red cell. An account of its chemical discrimination and pathology. Black well scientific publications. Oxford. - 1961.
7. Walensky L., Naria M., Lux, S. Disorders of the red blood cell membrane. Blood, principles and practice of hematology Ed. R.Mandin. - 2003.
A method for predicting the reduction of the transport function of the protein band 3 erythrocyte membranes newborn from mothers who have had an exacerbation of herpes virus infection in the third trimester of pregnancy, characterized by the fact that in the peripheral blood of a newborn to define the content of TNFα, identify in erythrocyte membranes newborn method disc electrophoresis in polyacrylamide gel (10%) sodium dodecyl sulfate protein strips-3, calculate the value of the discriminant functions by the formula
DF=(5.167 × strip-3)+(2.196×TNFα),
where DF is the discriminant function,
band-3 protein stroma membranes of erythrocytes,
TNFα - interleukin in the blood of newborns,
and when DF more 218.67 predict a decrease of the transport function of anions using a protein band-3 through erythrocyte membranes newborn.
SUBSTANCE: peripheral venous blood of the patients with autoimmune thyroiditis is examined for the absolute transferring receptor (CD71+) T-lymphocyte count and the interleukin-2 (IL-2) concentration. If the CD71+ T-lymphocyte count exceeds 0.3×109 cell/l, and the IL-2 concentration is less than 25 pg/ml, a favourable clinical course of the disease is predicted, while the CD71+ T-lymphocyte count less than 0.3×109 cell/l and the IL-2 concentration exceeding 25 pg/ml shows an unfavourable clinical course of autoimmune thyroiditis.
EFFECT: method provides more accurate and objective prediction with using the material, reagents and research methods widely applied in common clinical-laboratory practise.
SUBSTANCE: patient's peripheral blood is examined for the content of cytotoxic lymphocytes, %: (CD8/CD16), (CP8/GranzymeB), (CP16/GranzymeB), (CDS), (CD16/CD56) and (CD8/HLA DR). It is conmbined with determining cytotoxic activity (CTA), %. It is followed by calculating a RA activity index (AI) by formula: If observing the condition 11.8%<AI≤18.2%, a moderate degree of rheumatoid arthritis activity is diagnosed. If observing the condition AI>18.2%, a high degree of rheumatoid arthritis activity is diagnosed.
EFFECT: using the technique enables high-reliability differential diagnosis of the degree of rheumatoid arthritis activity.
SUBSTANCE: there are involved recording clinical and laboratory manifestations of a CNS injury during the first days of the disease that is followed by the calculation of total diagnostic coefficients related to the detected graduation levels of pathognomonic signs of the disease. Total (+)7 points and more is related to predicting the developing mixed tick-borne encephalitis and borreliosis infection. Total (-)8 points and less shows the developing potential monoinfection of tick-borne encephalitis. If deriving the intermediate values of total diagnostic coefficients when none of said limits is reached, the prognosis is uncertain.
EFFECT: using the method for stating basic tendencies of the developing pathological process at the early stages of the developing disease.
1 tbl, 2 ex
SUBSTANCE: with using indirect immunofluorescence and monoclonal antibodies, the patients suffering gastric ulcer (GU) are examined for an expression level of CD8, CDDR markers by peripheral blood lymphocytes with the CD8 level exceeding 24.5% and the CDDR level exceeding 15% enabling diagnosing a severe clinical course of GU.
EFFECT: use of the offered diagnostic technique provides high information value in diagnosing the severe clinical course of gastric ulcer, well-timed prediction of the severe clinical course of GU, instant diagnosing, provides well-timed and adequate complex of therapeutic measures.
1 tbl, 1 ex
SUBSTANCE: method involves the peripheral blood analysis for T-lymphocyte count with receptor expression in relation to CD3+ and CD25+ cells by cell count in 1 mcl of blood before the beginning of treatment and 10 days after. If observing the count increased 7% and more in comparison with the reference, a therapeutic effect in non-psychotic versions of psycho-organic syndrome is predicted to be positive.
EFFECT: use of the method enables more precise prediction of the clinical effectiveness in the given disease that promotes selecting an adequate therapy and reducing length of treatment.
SUBSTANCE: invention describes a method of detecting the decreased phosphatidyl ethanolamine content in red-cell membranes in a pregnant woman suffered aggravated herpes-virus infection in their third trimester of pregnancy with underlying increasing peripheral blood TNFα, characterised by the fact that peripheral blood of the pregnant woman is analysed for the TNFα content to detect phosphatidyl ethanolamine in red-cell membranes by two-dimensional thin-layer chromatography, and a discriminant equation is presented. Df=+1.743×TNFα+(-1.079×phosphatidyl ethanolamine), and if Df is more than a boundary value 78.86, the decreased phosphatidyl ethanolamine content in red-cell membranes is detected.
EFFECT: higher red-cell membrane stability in peripheral blood of the pregnant woman.
SUBSTANCE: invention describes a method for prediction of developing suppurative complications of acute pancreatitis involving laboratory blood analysis wherein the patients are examined for the kinetics of leukocyte chemiluminescence with using an activated suspension of clinical strains Acinetobacter baumannii or Pseudomonas aeruginosa or methicillin-resistant Staphylococcus aureus - MRSA and combining maximisation time - T max more than 1400.0 sec., maximum intensity - 1 max less than 5131.5 standard units, light sum - S less than 144000.0 standard units for the reaction initiated by Acinetobacter baumannii, and combining the values T max>1435.0 sec., I max<4708.0 standard units, S<118000.0 standard units for the reaction initiated by Pseudomonas aeruginosa, or combining the values T max>3520.0 sec., I max<4257.0 standard units, S<169000.0 standard units for the reaction initiated by MRSA enables predicting developing suppurative complications.
EFFECT: higher sensitivity and reduced time of prediction of suppurative complications of acute pancreatitis ensured by enabled assessment of the specific properties of patient's phagocytic response to etiologically significant activators.
1 ex, 4 tbl
SUBSTANCE: invention describes a method of assessment of the effect of arginine dehydrogenase activity on the blood histamine level in pregnant women suffered aggravated herpes viral infection with herpes antibody titre 1:12800 in the third trimester of gestation wherein syncytiotrophoblast of placental villi are examined with a histochemical reaction; section regions are photographed, entered in the computer and cytophotometried; observing increasing arginine dehydrogenase activity up to 48.7±2.9 standard units enables stating increasing the peripheral blood histamine level of the pregnant woman up to 1.44±0.09 mcmole/l that exacerbates foetoplacental deficiency in pregnant women.
EFFECT: method allows assessing the effect of arginine dehydrogenase activity on the blood histamine level of the pregnant women suffered aggravated herpes viral infection in the third trimester of gestation.
SUBSTANCE: pre-therapy blood plasma is analysed for immunoglobulin E, adrenocorticotrophin and endorphin, the index SCORAD dynamics is determined by formula: Dsk=1.34147-0.000820741*IgE-0.00382642*ACTP+0.0209401*endorphin wherein IgE is the level of acute blood plasma immunoglobulin E mIU/ml; ACTP is the level of acute blood plasma adrenocorticotrophin ng/l. If observing the Dsk value exceeding 1.18, a prognosis of improving clinical dynamics of recovery is favourable; the conventional therapy is considered to be sufficient. The Dsk value being equal to 1.18 or less, the improving clinical dynamics of recovery is unfavourable, the intensified therapeutic scheme for atopic dermatitis is considered to be preferential.
EFFECT: higher prediction accuracy.
SUBSTANCE: what is presented is a method for prediction of a character of an uterine involvement in myomatous nodes in a female patient suffering hysteromyoma involving DNA recovery from peripheral venous blood, analysis of +36 A/G locus genotypes of type 1 tumour necrosis factor receptor TKFR1 and analysis of myomatous nodes, and also measurement of an uterus. If detecting the genotypes +36 GG and +36 AG TNFR1, a lesser degree of the uterine involvement is predicted. The genotype +36 AA TNFR1 enables to predict a greater degree of the uterine involvement.
EFFECT: using the presented method allows predicting the character of the early uterine involvement in myomatous nodes.
4 dwg, 1 tbl, 1 ex
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.
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.
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
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.
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
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.