Plasmodium falciparum: Differential Sensitivity In Vitro to E-64 (Cysteine Protease Inhibitor) and Pepstatin A (Aspartyl Protease Inhibitor)

ABSTRACT We investigated the effect of a cysteine proteinase inhibitor (E-64) and an aspartyl proteinase inhibitor (Pepstatin A) on asexual erythrocytic stages of Plasmodium falciparum in culture. These two protease inhibitors showed different patterns of activity. E-64 acted preferentially against trophozoite and schizont stages. After 48 h incubation at high concentrations of E-64 (28, 140, 280 μM), growth was totally abolished and the parasites presented characteristic enlarged food vacuoles. Morphological alterations were also seen after shorter incubation periods (6 h at 28 μM) or 12 h at the inhibitory concentration 50% (12 μM), but an additional culture period (24 h) in inhibitor-free medium allowed normal parasite development, demonstrating a parasitostatic effect. E-64 acts on parasite multiplication; the normal merozoite maturation was altered and the normal reinvasion process partially impaired. Pepstatin A used at the inhibitory concentration 50% (4 μM) killed the parasites before trophozoite development and had a major effect on schizonts maturation. No altered parasite development occurred during an additional culture period without Pepstatin A, demonstrating a parasiticidal effect. E-64 and Pepstatin A used in combination inhibit the parasite growth with a strong synergistic effect.     

Device for the standard measurement of shoot flammability in the field

Fire ecology has been hindered by the lack of comparable, affordable protocols to quantify the flammability of whole plants over large numbers of species. We describe a low‐tech device that can be carried to the field and that allows highly standardized measurement of the flammability of whole individuals or portions up to 70 cm long. We illustrate its potential with results for 34 species belonging to different growth forms from central Argentina. The device consists of an 85 × 60 cm half‐cut metallic barrel placed horizontally and mounted on a removable metallic structure. It contains three parallel burners, a grill with an attached gauging thermometer and a blowtorch. Burners and blowtorch are connected to a propane–butane gas cylinder. Plant samples are placed on the grill and preheated with the burners for 2 min at 150°C. They are then ignited for 10 s with the blowtorch while the burners are kept on. Four parameters are measured for each sample: maximum temperature reached, burning time, burnt length and burnt biomass percentage. These parameters are used to construct a compound index of flammability for each sample that ranges between 0 (no flammability) and around 3 (maximum flammability). We obtained a wide range of values for flammability and all its components. Most of this variability was accounted for by differences between growth forms and species, rather than by differences at the level of replicates. This suggests that the device and protocol are sensitive enough to detect flammability differences among plants with different functional traits, and at the same time robust enough to produce consistent results among samples with similar traits. A major advantage is that plant architecture is kept almost intact, providing a flammability measure much closer to that of whole individuals in the field than those obtained by other standard protocols in use. The device and protocol presented here should facilitate the acquisition of comparable flammability data over large numbers of species from different floras and ecosystems, potentially contributing to several fields of research, such as functional ecology, evolutionary ecology and vegetation‐atmosphere modelling.