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Many investigators have suggested that sulfhydryl (SH) groups may be essential for the maintenance of the integrity of mammalian erythrocytes (1-4). This conclusion was drawn after observing the hemolytic effects of such heavy metals as mercurials and such oxidants as oxygen or iodine on erythrocytes. In hemolytic anemia in man, a relationship between hemolysis and a deficiency of reduced glutathione (GSH) in the red cells has been reported (5-7), however, there are discrepancies in the results (8-10). Jacob and Jandl (11) indicated that hemolysis induced by the oxidants and heavy metals resulted from a specific SH blockade, that is membrane, but not intracellular, SH blockade. However, there is apparently no report concerning the actual SH contents of the red cell membrane in relation to hemolysis. In the present work, various SH inhibitors were tested for their effects on SH levels of membrane and on hemolysis. Fresh human blood was centrifuged, the red cells were washed 3 times with phosphate buffered saline (154 mM NaCI, 9.6 mM Na,HPO4, 1.6 mM NaH2PO4, pH 7.4) and were then suspended to make 25 % hematocrit. One ml of buffered saline containing various amounts of SH inhibitor was added to 2 ml of the cell suspensions. The mixture samples were incubated for 1 hr at 37C and centrifuged at 10,000 rpm for 5 min at 4°C in a refrigerated centrifuge. The extent of hemolysis was measured by determining hemoglobin in the supernatant colorimetrically at 540 rim. After complete hemolysis with 30 ml of 15 mM Tris-HCI buffer (pH 7.4), the post hemolytic residue was washed 3 times with the same buffer and a milky membrane preparation was obtained. SH contents in the membrane fraction were estimated by the method described by ElIman (12). Protein content was determined by the method of Lowry et al. (13). Fig. I illustrates the relationship between the percent inhibition of membrane SH and degree of hemolysis induced by several heavy metals. Among the heavy metals tested , only mercurials were shown to have hemolytic activity. The highest activity was observed with HgCI2, followed in descending order by p-chloromercuribenzoate (PCMB) and methyl mercuric chloride (MMC). Regarding HgCI,, the percent hemolysis increased in proportion to the percent inhibition of membrane SH. The cause-effect relationship , however, could not be derived without integrated determinations of the results of other SH inhibitors. CdCl2 and CoCl,, at 1.6 mM inhibited membrane SH by almost 50% …

EFFECTS OF SAMMARIUM (Sm3+) ON THE CONTRACTILITY OF ISOLATED GUINEA PIG VENTRAL TAENIA COLI TO HISTAMINE