Monoamine oxidase-B (MAO-B) activity of platelets of an age- and sex-matched group of controls was compared with several groups of inpatients having non-familial dementia of Alzheimer type (DAT), Parkinson's disease (PD), multi-infarct dementia (MID), mixed types of these 3 diseases and a group of other central nervous system (CNS) organic disorders. All patients were subjected to several psychometric tests, including the Sandoz Clinical Assessment--Geriatric Scale, Hamilton Rating Scale for Depression, Mini-Mental State Examination and the Organic mental Disorder Scale (OMDS). A statistically significant enhancement of MAO-B activity could be observed in DAT patients and in PD patients, whereas the MID group showed a mean activity similar to that of the control group and the group with other organic CNS disorders. In DAT patients the degree of dementia in the OMDS test and the enhancement of MAO activity were positively correlated, but PD did not show such a correlation. It is concluded that the increase of MAO activity in PD and in DAT might be due to a disease-related enhanced affinity to oxygen and to such oxygen-derived radicals as superoxide or hydroxyl radicals. However, a possible drug-induced enhancement of MAO activity in PD cannot be excluded. Furthermore, the MAO-B activity values in platelets of individual patients or controls are not indicative of diagnosis or prognosis of any of these diseases and are of no disease-related specificity.
Treatment with cholesterol oxidases has shown that cholesterol is heterogeneously distributed in brush border membranes isolated from the apical domain of the renal and intestinal epithelial cells [Bloj, B., & Zilversmit, D. B. (1982) J. Biol. Chem. 257, 7608-7614; El Yandouzi, E. H., & Le Grimellec, C. (1992) Biochemistry 31, 547-551]. Cholesterol distribution between plasma membrane and intracellular membranes of the corresponding cells remains unexplored. The effects of Brevibacterium sp. cholesterol oxidase on the cholesterol content of LLC-PK1 cells, an epithelial cell line with multiple differentiated characteristics of the renal proximal tubule, were investigated. In confluent living cells grown as a monolayer on solid support, a small but significant fraction (13%) of the cholesterol was oxidized during the first hour of the oxidase treatment. Glutaraldehyde fixation prior to treatment resulted in a nearly complete (86.1 +/- 1.8) oxidation of the cellular cholesterol according to first-order kinetics. Filipin labeling and oxidation at 15 degrees C confirmed that cholesterol was essentially confined to the plasma membrane in LLC-PK1 cells. When adding the oxidase either on the apical or on the basolateral side of cells grown on permeant support and fixed with glutaraldehyde, a comparable monophasic oxidation of cholesterol was observed, despite the presence of efficient tight junctions. Adding the oxidase to both sides simultaneously did not increase the rate of oxidation. Finally, fixation of isolated renal brush border membranes with glutaraldehyde rendered undiscernible their cholesterol pools. We conclude that glutaraldehyde fixation, a commonly used process in the analysis of cholesterol distribution in cells, can mask the existence of cholesterol pools in plasma membranes.
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