Studies on the haemolytic action of amphiphilic substances in vitro. Inhibition by O-(beta-hydroxyethyl)-rutosides.
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Chemical Phenomena↗
Biomedical subjects
D B Wildenauer
Publications and source records attributed to D B Wildenauer.
Reactions of the alkylating agent tris(2-chloroethyl)-amine with the erythrocyte membrane. Effects on shape changes of human erythrocytes and ghosts.
The influence of tris(2-chloroethyl)amine on shape changes of human erythrocytes and ghosts was studied in vitro and correlated with alterations in the molecular structure of the membrane. (1) Reaction with 1--2 mM tris(2-chloroethyl)amine, a concentration which caused polymerisation of spectrin as detected by sodium dodecyl sulphate polyacrylamide gel electrophoresis, protected intact erythrocytes against metabolically induced shape changes. (2) When induced by Mg2+-ATP, ghosts porepared from alkylated erythrocytes underwent normal changes in shape. However, when ghosts were treated directly with tris(2-chloroethyl)amine, no Mg2+-ATP-induced shape changes occurred. This fixation in shape appeared to be due to a higher degree of reaction with the alkylating agent. (3) The amount of chlorpromazine necessary for transformation of erythrocytes into stomatocytes was increased for tris(2-chloroethyl)amine-pretreated cells and was dependent on the degree of reaction with tris(2-chloroethyl)amine. (4) Deformability of red cells after reaction with tris(2-chloroethyl)amine was estimated by measuring their rheological behaviour in glass capillary arrays. A slight reduction of the flow rate was observed for cells alkylated with 1--2 mM tris(2-chloroethyl)amine. (5) Extractability of spectrin was diminished and corresponded to the degree of spectrin polymerisation. (6) No difference in the incorporation of 32P by alkylated and untreated cells was found. Sodium dodecyl sulphate polyacrylamide gel electrophoresis and autoradiography of the membrane proteins from 32-P-treated cells showed that the spectrin component 2 and the polymerisation products generated by the reaction with tris(2-chloroethyl)amine were labelled. It is suggested that the observed conservation of cell shape is preferentially caused by the reaction of tris(2-chloroethyl)amine with spectrin.
Alkylating Agents↗
Analysis of cerebrospinal fluid from patients with psychiatric and neurological disorders by two-dimensional electrophoresis: identification of disease-associated polypeptides as fibrin fragments.
Two-dimensional electrophoresis (2-DE) of cerebrospinal fluid (CSF) samples--from 347 patients with various psychiatric and neurological disorders--and subsequent silver staining revealed two additional polypeptides (Mr 40,000) in 49% of 111 schizophrenics, 46% of 43 schizoaffective patients, 36% of 41 patients with affective disorders, 43% of 28 patients with multiple sclerosis, but not in 25 patients without neurological symptomatology, nor in 9 patients with Lues, and in only 2 of 25 patients with AIDS. The two polypeptides, as detected by 2-DE, eluted after size exclusion chromatography in fractions containing proteins with Mr greater than 200,000. After 2-DE of CSF samples, enriched by gel chromatography, the polypeptides were immobilized by blotting onto glass-fiber membranes and subjected to N-terminal sequencing. Polypeptide A was identified as beta-chain remnant (beta 2), derived from plasmin cleavage of fibrin(ogen). After size exclusion chromatography, 2-DE, and Western blotting, polypeptide A and B, as well as several other spots, reacted with fibrinogen antibodies, suggesting that the polypeptides are subunits of a fibrin degradation complex.
Acquired Immunodeficiency Syndrome↗