Method for prediction of erythrocyte membrane glycophorin activity in peripheral blood of pregnant women suffering acute condition of herpesviral infection and fatty acid peroxide concentration in peripheral blood

FIELD: medicine.

SUBSTANCE: patient's blood is analysed for the erythrocyte membrane glycophorin concentration by gel electrophoresis assay and the malonic aldehyde concentration by spectrophotometric analysis. Then a discriminant equation D =-2.719 · glycophorin +{+25.331 · malonic aldehyde} is solved. If D is equal to or exceeds a boundary value +9.52, higher glycophorin activity in erythrocyte membranes is predicted in pregnant women.

EFFECT: higher prediction accuracy.

2 tbl, 3 ex

 

The purpose of the invention consists in predicting the degree of activity of glycophorin membranes of erythrocytes in the peripheral blood of pregnant women during exacerbation of herpes viral infections.

Glycophorin is a major integral membrane protein of the erythrocyte, bearing on its surface a negative charge, which contributes to the repulsion of erythrocytes from each other. In addition, glycophorin is a receptor for a number of antigens and depending on contact with one of them appear certain morphological and functional properties of erythrocytes (Dranton et al, 2001; Gallgher P.G., Forget B.G, 2001; Bemnet V., 1985; Vorob'ev A.I., 2007). In the existing literature have no coverage issues activity glycophorin during pregnancy, complicated by herpes virus infection. To refer to any prototype, we formulate our aspect, it is not possible.

The lack of data allows us to formulate the forecast activity of integral membrane protein of erythrocytes glycophorin during exacerbation of herpes viral infection during pregnancy.

The objective is achieved by the fact that the evaluation activity glycophorin in the membranes of erythrocytes in peripheral blood of pregnant women during exacerbation of herpes viral infection is determined depending on the accumulation of end products of peroxides fatty acid - malondialdehyde.

The claimed method ameeshaaeysha techniques:

a) using disc electrophoresis in a stepwise density gradient polyacrylamide gel (10%) sodium dodecyl sulfate determine the protein fraction of erythrocyte membranes, including glycophorin

b) determination of malondialdehyde (MDA) is performed using thiobarbituric acid on spectrophotometer "Stat-Fax-1900" (USA) [2].

b) the titer of antibodies to herpes simplex determine enzyme-linked immunosorbent assay in microplate analyzer, "Stat Fax-2100" (USA) using sets of "Vector-best" (Novosibirsk).

g) evaluating the relationship between the studied parameters in healthy pregnant women and pregnant women who have had an exacerbation of herpes viral infection, solve using discriminant equation D=-2,719 · glycophorin + {+25,331-malonic dialdehyde} (PL. 2). Malonic dialdehyde selected as the most relevant variable in multiple linear regression (table. 1).

On the basis of discriminant analysis

D=-2,719·8,9+{+25,331·1,65}

D=-21,99+{+41,79}=+19,80

D(+19,80)>the boundary values (+9,52)

and with the increasing content of malondialdehyde in the peripheral blood of pregnant women during exacerbation of herpes-virus infection (antibody titer of 1:12800) predict an increase in the activity of glycophorin erythrocyte membrane.

d) to illustrate the inventive method examples (1-3).

Literature

1. Vorob'ev A.I. the owner on Hematology. - M.: Novamed, 2007. - s.

2. Springing I.D., Garishvili YEAR the Method of determining the dialdehyde with thiobarbiturate acid: Modern methods in biochemistry. - M.: Medicine, 1977. - 390 S.

3. Bemnet V. The membrane skeleton of human erythrocyte implications for more complex cell / Annu. Rev. Biochem. - 1985. - Vol.54. - P.273-304.

4. Branton D., Coher S.M., Tyler J. Interaction of cytoskeletal on the human erythrocyte membrane / Cell. - 1981. - Vol.24. - P.22-32.

5. Gallgher P.G., B.G. Forget The ret cell membrane. Williams Hematology. / Edit. E. Beutler, M. Lichtman, B.S. Cooler et al. - Charter 45, 6th edition. Mc. Grow Hill International Edition, 2001. - C.333-343.

EXAMPLES

Example 1. Pregnant A., 20 years. Moved in the third trimester exacerbation of herpes viral infection with antibody titer of 1:12800. In peripheral blood was determined by a spectrometric method (Stat-Fax-1900; USA) 30,51 µmol/l of malonic dialdehyde. Using disc electrophoresis in a stepwise density gradient polyacrylamide gel (10%) sodium dodecyl sulfate was determined protein fraction of erythrocyte membranes, including glycophorin. The percentage of glycophorin was 12.0 percent.

Discriminant equation in this case shows: D=-2,719·12+{+30,51·1,65}=16,86, that is, it is greater than the boundary value (+9,52). This pregnant predict an increase of glycophorin to 12.0% compared with control (6,65±0,08%) content in peripheral blood malondialdehyde 30,51 µmol/l during exacerbation of herpes viral infections.

Example 2. Pregnant,, 23 years. Moved the third trimester exacerbation of herpes viral infection with antibody levels - 1:12800. In peripheral blood was determined by a spectrometric method (Stat-Fax-1900; USA) 36,51 µmol/l of malonic dialdehyde. Using disc electrophoresis in a stepwise density gradient polyacrylamide gel (10%) sodium dodecyl sulfate was determined protein fraction of erythrocyte membranes, including glycophorin. The percentage of glycophorin was 13,0±1,2%. D=-2,719·13+{+36,51-1,65}=24,90. Boundary value (+9,52).

This pregnant when 36,51 µmol/l of malonic dialdehyde in the peripheral blood during exacerbation of herpes viral infection predict an increase in the percentage content in erythrocyte membranes of glycophorin to 13.0±1,2%) compared with control (6,15±0,08%).

Example 3. Pregnant With, 24 years. In the third trimester have not suffered any diseases. The amount of malondialdehyde and peripheral blood was determined by a spectrophotometric method (Stat-Fax-1900; USA) 11,33 µmol/L. using disc electrophoresis in a stepwise density gradient polyacrylamide gel (10%) sodium dodecyl sulfate was determined protein fraction of erythrocyte membranes, including glycophorin. The percentage of glycophorin was 7.5 percent.

Discriminant equation in this case shows: D=-2,719·7,5+{+11,33-1,65}=2,67. That is, it is less than the boundary value (+9,52).

This pregnant do not predict an increase in glycophorin above the boundary values (6,65±0,08%) for the content of malondialdehyde within 11,33 µmol/l and no exacerbation of herpes-virus infection in the third trimester.

Table 2
Automated screening system.today 07.12.2008 22:53
Statistical processing of resultsdiscriminant analysis
Name sampleSampleThe number of parametersDistance MaharanaThe Fisher Test. Differences sampleThe probability differences of samples %Boundary value of the discriminant functionsThe probability of a wrong class %
4931424,6563,80100,0009,52is 1,809
OOR-control112 1,365
D=-2,719·8 glycophorin + 25,331 malonic dialdehyde (MDA)
Note: 493 - sampling with a herpes viral infection; OOR - control.

D=-2,719·8,9+{+25,331·1,65}

D=-21,99+{+41,79}=+19,80

D(+19,80)>the boundary values (+9,52)

A method for predicting activity of glycophorin membranes of erythrocytes in the peripheral blood of pregnant women during exacerbation of herpes virus infection and the increase of peroxides fatty acids in the blood by determining the content of glycophorin membranes of erythrocytes by the method of gel-eletrophoresis and malondialdehyde spectrophotometric, characterized by the fact that decide discriminant equation D=-2,719 · glycophorin +{+25,331 · malonic aldehyde}, and for D equal to or greater than the boundary values +9,52, predict an increase in the activity of glycophorin in erythrocyte membranes in pregnant women who have had an exacerbation of herpes virus infection.



 

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