thiobarbituric-acid and Myocardial-Ischemia

It was established that viral particles, like low-density lipoproteins (LDLP), when subjected to some modification changes, lost their ability to be internalized by tissue somatic cells and acquired tropism to macrophage cells. The data, obtained by us by using the polymerase chain reaction (PCR) method, made it possible to assert that atherosclerotic plaques, isolated from vessels of patients with ischemic heart disease (IHD) who underwent coronary bypass, contained RNA of the A(HINI) and AH3N3) influenza viruses. Whereas, the vessel portions, undamaged by atherosclerosis, did not contain any genetic substances of influenza viruses. It was for the first time that an experimentally supported understanding was expressed on that the atherosclerotic plaques serve as a "reservoir" for influenza viruses. It is also suggested that the mentioned plaques can be the carriers of influenza viruses for a long time, thus, prolonging the persistent form of influenza infection in the human body.

Topics: Aged; Animals; Arteriosclerosis; Coronary Artery Bypass; Coronary Artery Disease; Coronary Vessels; Humans; Influenza A virus; Influenza, Human; Lipid Peroxidation; Lung; Malondialdehyde; Mice; Middle Aged; Myocardial Ischemia; Polymerase Chain Reaction; RNA, Viral; Spectrophotometry; Thiobarbiturates; Time Factors; Tropism

The involvement of oxidatively modified low density lipoprotein (LDL) in the development of CHD is widely described. We have produced two antibodies, recognizing the lipid oxidation product malondialdehyde (MDA) on whole LDL or ApoB-100. The antibodies were utilized in the development of an ELISA for quantitation of MDA-LDL in human plasma. Intra- and inter-assay coefficients of variation (% CV) were measured as 4.8 and 7.7%, respectively, and sensitivity of the assay as 0.04 micro g/ml MDA-LDL. Recovery of standard MDA-LDL from native LDL was 102%, indicating the ELISA to be specific with no interference from other biomolecules. Further validation of the ELISA was carried out against two established methods for measurement of lipid peroxidation products, MDA by HPLC and F(2)-isoprostanes by GC-MS. Results indicated that MDA-LDL is formed at a later stage of oxidation than either MDA or F(2)-isoprostanes. In vivo analysis demonstrated that the ELISA was able to determine steady-state concentrations of plasma MDA-LDL (an end marker of lipid peroxidation). A reference range of 34.3 +/- 8.8 micro g/ml MDA-LDL was established for healthy individuals. Further, the ELISA was used to show significantly increased plasma MDA-LDL levels in subjects with confirmed ischemic heart disease, and could therefore possibly be of benefit as a diagnostic tool for assessing CHD risk.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Animals; Apolipoproteins B; Case-Control Studies; Copper; Enzyme-Linked Immunosorbent Assay; F2-Isoprostanes; Female; Gas Chromatography-Mass Spectrometry; Humans; Immunoglobulins; Lipid Peroxidation; Lipoproteins, LDL; Male; Malondialdehyde; Mice; Mice, Inbred BALB C; Middle Aged; Myocardial Ischemia; Oxidation-Reduction; Rabbits; Risk Factors; Thiobarbiturates

Estimation of lipid peroxidation (LPO) through malonaldehyde (MDA) formation measured by assaying thiobarbituric acid reactive products remains the method of choice to study the development of oxidative stress to assess myocardial ischemic reperfusion injury. However, MDA estimation by this assay is non-specific and often gives erroneous results. In this report, we describe a method to estimate MDA, formaldehyde (FDA), acetaldehyde (ADA), and acetone, the degradation products of oxygen free radicals (OFR) and polyunsaturated fatty acids (PUFA), as presumptive markers for LPO. Isolated rat hearts were made ischemic for 30 min, followed by 60 min of reperfusion. The perfusates were collected, derivatized with 2,4-dinitrophenylhydrazine, and extracted with pentane. Aliquots of 25 microliters in acetonitrile were injected on a Beckman Ultrasphere C18 (3 microns) column. The products were eluted isocratically with a mobile phase containing acetonitrile-water-acetic acid (40:60:0.1, v/v/v). The peaks were identified by co-chromatography with the hydrazine derivatives of authentic standards. The retention times of MDA, FDA, ADA and acetone were 5.0, 6.3, 9.8 and 15.7 min, respectively. The results of our study indicated progressive increase in all four lipid metabolites with reperfusion time. Thus, our results demonstrate that the release of lipid metabolites from the isolated heart increased in response to oxidative stress. Since MDA, FDA, ADA, and acetone are the products of OFR-PUFA interactions, this method allows proper estimation of LPO to monitor the oxidative stress developed during the reperfusion of ischemic myocardium.

Topics: Animals; Calibration; Chromatography, High Pressure Liquid; In Vitro Techniques; Lipid Metabolism; Lipid Peroxidation; Male; Malondialdehyde; Myocardial Ischemia; Myocardial Reperfusion; Rats; Rats, Sprague-Dawley; Spectrophotometry, Ultraviolet; Thiobarbiturates

Previous studies demonstrated that preconditioning of a heart by repeated stunning can reduce the cellular injury to the heart from subsequent acute ischemic insult. To examine the possible biochemical mechanism for such myocardial preservation afforded by preconditioning, swine heart was subjected to four episodes of 5 min. stunning by occluding the left anterior descending coronary artery (LAD), followed by 10 min. of reperfusion after each stunning. Heart was then made regionally ischemic for 60 min. by LAD occlusion, followed by 6 hrs. reperfusion. Control heart was perfused for 60 min., followed by 60 min. ischemia and 6 hrs. reperfusion. The results of our studies indicated the stimulation of a number of antioxidative enzymes, including Mn-superoxide dismutase (Mn-SOD), catalase, glutathione peroxidase, and glutathione reductase, after repeated stunning and reperfusion. In addition, a number of new proteins were expressed after preconditioning the heart, including some oxidative-stress related proteins and 72 kDa heat-shock protein. These results suggest that preconditioning of a heart by repeated stunning may lead to strengthening of the oxidative defense system of the heart, which is likely to play a role in myocardial preservation during subsequent ischemic and reperfusion injury.

Topics: Adaptation, Physiological; Animals; Antioxidants; Catalase; Electric Stimulation; Electrophoresis, Gel, Two-Dimensional; Glutathione Peroxidase; Glutathione Reductase; Heart; Heat-Shock Proteins; Myocardial Ischemia; Myocardial Reperfusion; Myocardium; Superoxide Dismutase; Swine; Thiobarbiturates