Calpains and cathepsin D of the injured myocardium: Role of iodothyronines

Stroev, E. A.; Ryazanova, E. A.

doi:10.1007/bf00840463

Cited by 1 publication

(1 citation statement)

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“…The supernatant was used to determine the activity of μ-calpain with the release of enzyme using ion exchange chromatography (column 3 × 15; DEAE-cellulose) and eluted by a gradient of 0.1–0.4 M NaCl. Next, the mean calpain activity of the fractions was determined using incubation medium previously described by Sorimachi et al ( 1997 ) (imidasole buffer, 4% casein, 50 mM of CaCl 2 , 50 mM of cysteine) (Stroev et al, 1991 ; Elce John, 2000 ; Sorimachi et al, 2000 ). Then, the general proteolytic activity, in ascending order of the absorption level at a wavelength of 280 nm, was measured following incubation of the haemolysate and protein deposition by 5% trichloroacetic acid (TCA) (Bazarnova et al, 2008 ).…”

Section: Methodsmentioning

confidence: 99%

Physical and Chemical Processes and the Morphofunctional Characteristics of Human Erythrocytes in Hyperglycaemia

Ревин

Klenova²,

Gromova

et al. 2017

Front. Physiol.

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Background: This study examines the effect of graduated hyperglycaemia on the state and oxygen-binding ability of hemoglobin, the correlation of phospholipid fractions and their metabolites in the membrane, the activity of proteolytic enzymes and the morphofunctional state of erythrocytes.Methods: Conformational changes in the molecule of hemoglobin were determined by Raman spectroscopy. The structure of the erythrocytes was analyzed using laser interference microscopy (LIM). To determine the activity of NADN-methemoglobinreductase, we used the P.G. Board method. The degree of glycosylation of the erythrocyte membranes was determined using a method previously described by Felkoren et al. Lipid extraction was performed using the Bligh and Dyer method. Detection of the phospholipids was performed using V. E. Vaskovsky method.Results: Conditions of hyperglycaemia are characterized by a low affinity of hemoglobin to oxygen, which is manifested as a parallel decrease in the content of hemoglobin oxyform and the growth of deoxyform, methemoglobin and membrane-bound hemoglobin. The degree of glycosylation of membrane proteins and hemoglobin is high. For example, in the case of hyperglycaemia, erythrocytic membranes reduce the content of all phospholipid fractions with a simultaneous increase in lysoforms, free fatty acids and the diacylglycerol (DAG). Step wise hyperglycaemia in incubation medium and human erythrocytes results in an increased content of peptide components and general trypsin-like activity in the cytosol, with a simultaneous decreased activity of μ-calpain and caspase 3.Conclusions: Metabolic disorders and damage of cell membranes during hyperglycaemia cause an increase in the population of echinocytes and spherocytes. The resulting disorders are accompanied with a high probability of intravascular haemolysis.

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Section: Methodsmentioning

confidence: 99%