Stage-Dependent Effects of Chloroquine on Plasmodium falciparum In Vitro1

The erythrocytic developmental cycle of Plasmodium falciparum can be conveniently divided into the ring, trophozoite, and schizont stages based on morphology and metabolism. Using highly synchronous cultures of P. falciparum, considerable variation was demonstrated among these stages in sensitivity to chloroquine. The effects of timed, sequential exposure to several clinically relevant concentrations of chloroquine were monitored by three techniques: morphological analysis, changes in the rate of glucose consumption, and changes in the incorporation of ³H-hypoxanthine into parasite nucleic acids. All three techniques gave essentially identical results. The trophozoite and schizont stages were considerably more sensitive to the drug than ring-stage parasites. Chloroquine sensitivity decreased as nuclear division neared completion. The increase in chloroquine sensitivity was coincident with a marked rise in the rate of glucose consumption and nucleic acid synthesis. The rate of nucleic acid synthesis decreased as schizogony progressed while glucose consumption continued at high rates during this process. The degree of chloroquine sensitivity was not highly correlated with either metabolic activity.     

Bean dwarf mosaic virus: a model system for the study of viral movement

Taxonomy: Bean dwarf mosaic virus‐[Colombia:1987] (BDMV‐[CO:87]) is a single‐stranded plant DNA virus, a member of the genus Begomovirus of the family Geminiviridae. Physical properties: BDMV virions are twinned incomplete isosahedra measuring 18 × 30 nm. The viral particle is composed of 110 subunits of coat protein, organized as 22 pentameric capsomers. Each subunit has a molecular mass of ～29 kDa. BDMV possesses two DNA components (designated DNA‐A and DNA‐B), each ～2.6 kb in size. Host range: The natural and most important host of BDMV is the common bean (Phaseolus vulgaris). Nicotiana benthamiana is often used as an experimental host. Common bean germplasm can be divided into two major gene pools: Andean materials, which are mostly susceptible to BDMV, and Middle American materials, which are mostly resistant to BDMV. Disease symptoms: The symptom intensity in common bean plants depends on the stage of infection. Early infection of susceptible bean seedlings will result in severe stunting and dwarfing, leaf distortion and mottling or mosaic, as well as chlorotic or yellow spots or blotches. BDMV‐infected plants usually abort their flowers or produce severely distorted pods. Late infection of susceptible plants or early infection of moderately resistant genotypes may show a mild mosaic, mottle and crumpling or an irregular distribution of variegated patches. Biological properties: As a member of the Begomovirus group, BDMV is transmitted from plant to plant by the whitefly Bemisia tabaci. BDMV is a nonphloem‐limited virus and can replicate and move in the epidermal, cortical and phloem cells. As a nonphloem‐limited virus, it is sap‐transmissible.     

Comparative Effects of Soil Moisture Stress and Restricted Root Zone Volume on Morphogenetic and Physiological Responses of Soybean [Glycine max (L.) Merr.]

Significant differences in response to soil moisture stress(SMS) and restricted root zone volume (RRZV) were found in twocultivars of soybean [Glycine max (L.) Merr.] (‘Forrest’and ‘Williams’) plants grown under controlled-environmentconditions. Leaf water potentials of SMS-treated plants were0·4-0·6 MPa lower than those of controls and stomata1conductances 23-56% lower. In the case of RRZV treatment, however,there were no differences in either parameter. Initiation ofnew leaves as reflected in the plastochron index was stronglyreduced by SMS but was unaffected by RRZV. Photosynthetic rates(CO₂ fixation dm² of leaf) of plants given SMS were reducedby 11-21% while those of RRZV-treated plants were unaffected.SMS caused a strong preferential allocation of dry matter tothe root at the expense of the shoot in both cultivars. RRZV,however, had no effect on assimilate distribution in ‘Forrest’and only slightly favoured root growth in ‘Williams.’Carbohydrate concentrations of both alcohol-soluble and insolublefractions were increased significantly by SMS, especially inthe leaves, but were little affected by RRZV. Nitrogen concentrationin the root fraction was reduced by 22-24% and that in the leafand stem fractions by 7-14% under SMS but was not affected appreciablyby RRZV. Phosphorus concentration in the leaf, stem, and rootfractions was reduced by 45-65% under SMS but was relativelyunaffected by RRZV. These findings suggest that SMS and RRZVare basically different in their mechanism of action and thatthe impairment of growth resulting from these two stresses mayinvolve different physiological processes. Our results alsoindicate that the suppressive effects of small containers onplant growth do not necessarily result from inadvertent SMS. Key words: Drought, Container effects, Glycine max (L.) Merr     

Effects of Chloroquine on the Feeding Mechanism of the Intraerythrocytic Human Malarial Parasite Plasmodium falciparum1

Ultrastructural investigations of P. falciparum cultivated in vitro in human erythrocytes revealed new features of the feeding mechanism of the parasite. Mature trophozoites and schizonts take up a portion of the host cytosol by endocytosis which is restricted to cytostomes and which involves the invagination of both parasitophorous and parasite membranes. The resulting endocytic vesicles, surrounded by two concentric membranes, migrate towards the central food vacuole membrane. The external membrane of the endocytic vesicles apposes that of the food vacuole, leading to the internalization of vesicles bounded by a single membrane into the vacuolar space where they are rapidly degraded. We conclude from this sequence of events that endocytic vesicles fuse with the food vacuole. Treatment of infected cells with therapeutic concentrations of chloroquine inhibited the last step of the feeding process, i.e. vacuolar degradation. This was manifested by the accumulation within the vacuolar space of intact vesicles bounded by single membranes. The implications of these findings for the antimalarial activity of chloroquine are discussed.     

Effects of Mitochondrial Protein Synthesis Inhibitors on the Incorporation of Isoleucine into Plasmodium falciparum In Vitro1

Plasmodium falciparum was grown in human erythrocytes in vitro and the effect of chloramphenicol, erythromycin, and tetracycline on growth and maturation of the parasites and on their ability to incorporate [³H]isoleucine into protein was observed. Exposure of rings to high concentrations of chloramphenicol had little effect on subsequent maturation of the rings whereas brief (4 h) exposure of trophozoites caused a dose-dependent inhibition of subsequent ring formation. Incorporation of [³H]isoleucine into protein was not affected during at least 6 h of exposure to high concentration of the three drugs examined, but appreciable inhibition was observed after 21 h, with chloramphenicol being the least effective inhibitor. These results suggest that there is a stage-specific effect of inhibition of mitochondrial protein synthesis on subsequent development and that the mitochondria are essential for growth and development even though they lack a functional Krebs cycle.