Initial extracellular development in vitro of merozoites of Plasmodium falciparum

Late schizonts from continuous cultures of P. falciparum were concentrated over Percoll, inoculated to various experimental media at the rate of about 20 X 10(6) per 0.5 ml of medium, and incubated in a candle jar at 37 degrees for 1 day. Controls in standard culture medium showed a heavy invasion with young rings in the previously uninfected red cells introduced with the inoculum of schizonts. In a medium of high potassium content containing a 33% extract of human erythrocytes, this invasion was inhibited and many free merozoites were present. If, however, this same medium was supplemented with both ATP, as the dipotassium salt at 1.6 mM, and sodium pyruvate at 3.6 mM, there appeared large numbers of extracellular forms resembling young rings. Examination of these by electron microscopy shows that they are indeed merozoites that have begun to differentiate extracellularly. This suggests that the trigger for differentiation of merozoites may not depend on the process of entry into a red cell but rather on specific factors within the red cell.     

NAG infection in Rana temporaria with suppressed normal intestinal microflora]

Frogs of the Rana temporaria species with disturbed biocenosis induced by combined effects of hypothermia (+4 degrees C) and tetracycline were used as an experimental model. The animals were inoculated orally with cultured NAD-vibrios and subjected to clinical, bacteriological, immunomorphological and electron microscopical examinations. The lymph-blood system was shown to be a possible pathway for generalization of the infection. Several causes of long-term persistence of the vibrios in Rana temporaria are discussed.     

A sensitive and specific stable isotope assay for warfarin and its metabolites

A capillary gas chromatographic mass spectrometric method for the quantification of warfarin and its known metabolites from microsomal incubations is described. Deuterium labelled 4', 6-, 7- and 8-hydroxy warfarins are used as internal standards and the method has detection limits of 1 ng ml-1 with 20 ng ml-1 being the lower limit for accurate quantification.     

Plasmodium falciparum in culture: improved continuous flow method.

A new design of flow vessel provides a method for continuous culture of P. falciparum in a settled layer of human erythrocytes with a slow flow of culture medium over them. The parasitemia is kept fluctuating from approximately 1%, just after addition of fresh erythrocytes, to approximately 10%, 2 or 3 days later. Each vessel provides each week 3 harvests, each containing approximately 0.6--1 X 10(9) parasites.