Department of Biochemistry, Sri Krishnadevaraya University, Anantapur, India
* Author to whom correspondence should be addressed at: Department of Biochemistry, Sri Krishnadevaraya University, Anantapur 515003, India. E-mail: nchvaradacharyulu{at}yahoo.com
(Received 6 June 2003; first review notified 22 August 2003; in revised form 1 December 2003; accepted 15 December 2003)
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ABSTRACT |
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INTRODUCTION |
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MATERIALS AND METHODS |
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Sampling and assay methods
Freshly drawn heparinized blood samples were used for membrane and plasma analysis. Erythrocyte membranes were prepared as per the method of Dodge et al. (1963); membrane protein was estimated following the method of Lowry et al. (1951)
. Extraction of lipids from erythrocyte membrane and the determination of cholesterol and phospholipids were done as per the method adopted by Jain et al. (1988)
. The plasma cholesterol was assayed according to the enzymatic kit method of Allian et al. (1974)
. Plasma phospholipids were assayed according to the method of Connerty et al. (1961)
.
To determine membrane fluidity, the optical method of Galla and Sackmann (1974), as adopted by Bryszewska et al. (1986)
, was used. This method was based on the dependence of microviscosity the lateral diffusion rate of fluorescent aromatic hydrocarbon pyrene. A measure of lateral diffusion rate is the ratio of fluorescence intensity of dimer and monomer (D/M ratio) in the fluorescence spectrum of pyrene. Pyrene fluorescence was excited at 339 nm and the intensities were measured at 395 nm (monomer) and 469 nm (dimer). All the fluorescence measurements were performed at 37°C with an F-3010 fluorescence spectrophotometer (Hitachi, Japan). Red cell preparation was done according to the method described by Beutler (1975)
and the extent of lipid peroxidation in red cells was measured by assaying the malondialdehyde (MDA) formed (Buege and Aust, 1978
).
All the chemicals were of analytical grade and were from Sigma Chemical (St. Louis, MO), Spectrochem (Mumbai, India) or Qualigens Fine Chemicals (Mumbai, India).
Statistical analysis
The data obtained in the present study were subjected to Student's t-test.
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RESULTS |
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DISCUSSION |
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ACKNOWLEDGEMENTS |
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REFERENCES |
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