Incubation of purified rat kidney mitochondrial fraction with phospholipase-D resulted in the accumulation of phosphatidic acid in the membrane due to the degradation of membrane-bound phosphatidylcholine, -serine and-ethanolamine Simultaneously with the hydrolysis of the phospholipids, cholesterol and protein were released from the mitochondrial membrane into the medium, and binding of Ca²⁺ by mitochondrial membranes increased. Phospholipase Dtreated mitochondrial fraction exhibited increased swellingin vitro in the early stages of incubation (15 min) after which the mitochondria were ruptured. Membrane-bound adenosine triphosphatase was partially inactivated and the enzyme activity was not significantly restored by incubation with sonicated dispersions of phosphatidylcholine,-serine and cholesterol. These results indicate that removal of choline, serine and ethanolamine from membrane-bound phospholipids disrupt phospholipid-cholesterol and phospholipid-protein association and affect functions of the membrane.

Addition of sonicated dispersions of cholesterol to peptone-salt-vitamin medium resulted in the metabolism of the sterol byHartmanella culbertsoni. Trophozoite multiplication was stimulated at 1–5 mg/litre, but retarded at 10–20 mg/litre. When cholesterol was added to the medium, incorporation of [1,2-¹⁴C] -acetate into neutral lipid, phospholipid, non-saponifiable and cholesterol fractions of the amoebae was significantly reduced. Cholesterol ester was detected in the medium but phospholipids were not released. Addition of cholesterol stimulated the activity of lysosomal acid phosphatase, acid deoxyribonuclease and cathepsin B but did not affect 5′-nucleotidase, adenosine triphosphatase, alkaline phosphatase, glucose-6-phosphatase, succinate dehydrogenase and cytochrome C oxidase.

Fatty acids, cholesterol and glucose present in axenic medium are utilized by growingEntamoeba histolytica but the amoeba is unable to synthesize cholesterol from [U^-14 C^- ] glucose although the label is incorporated into the fatty acids and non-saponifiable fractions of the organism. Exogenously-added sonicated dispersions of cholesterol, Β-sitosterol, lanosterol, lecithin and lauric, palmitic, linoleic and stearic acids are ingested by the amoebae with subsequent loss in amoeboid movement. After a few hours the movement is regained. Cholesterol, lecithin and the fatty acids stimulate amoebic multiplication but are unable to replace serum in the medium either singly or in combination.

A rapid method for preparation of plasma membrane fromAcanthamoeba culbertsoni involving toluene treatment followed by lithium bromide extraction is described. In the plasma membrane preparation, 5′-nucleotidase, Na⁺ + K⁺ -ATPase, Mg²⁺ -ATPase and glucose-6-phosphatase activities were enriched. The membrane preparation was free from nucleic acid, cytochrome P-450 and cytochrome b5. Amino acid (¹⁴C-Ieucine) was not incorporated in the plasma membrane in 2 min. Succinic dehydrogenase was not detectable in the plasma membrane preparation. The molar ratio of cholesterol and phospholipids was 0.95 which is characteristics for plasma membranes. Under electronmicroscopy the preparation was homogenous without any other component of the cell. Plasma membrane proteins and glycoproteins were separated on acrylamide gel electrophoresis

Levels of lipid peroxides in rat caecum, blood, liver and kidney and the capacity of tissue homogenates to form lipid peroxidesin vitro was enhanced after caecal amoebiasis in rats produced byEntamoeba histolytica (IB-1). The activity of hepatic drug-metabolizing enzymes in post-mitochondrial fraction and the cytochrome P₄₅₀ contents in microsomal fraction decreased significantly, while lysosomal enzymes such as acid phosphatase, acid ribonuclease and cathepsin B showed an increase in the liver homogenates of infected animals. These changes were reversed following treatment with the antiamoebic drug, metronidazole