Effect of Zinc Deficiency on Proton Fluxes in Plasma Membrane-Enriched Vesicles Isolated from Bean Roots

Plasma membrane-enriched vesicles were isolated by density gradientcentrifugation from roots of zinc-sufficient and zinc-deficientbean (Phaseolus vulgaris L. cv. Prélude) plants. Thetwo populations of vesicles had similar activities of specificmembrane marker enzymes and ATP hydrolysis and were competentfor proton transport. However, vesicles from zinc-deficientroots showed lower rates of ATP-dependent intravesicular acidificationand increases in passive permeability to protons as well asin the rate, of dissipation of a non-metabolic transmembranepH gradient. The decrease of the rate of proton accumulationin isolated vesicles closely paralleled the increase in potassiumleakage from intact roots and the appearance of visual zincdeficiency symptoms in the shoots. Re-supply of zinc to deficientplants for 24 h promoted shoot growth, reduced potassium leakagefrom roots and led to partial recovery of the proton accumulationcapacity and to a decrease in passive permeability to protonsin isolated vesicles. The results obtained with isolated vesiclesconfirm the previously observed ‘in vivo’ effectsof zinc deficiency and are consistent with the idea that analteration of plasma membrane lipids leads to an increase inpermeability and an impairment in trans-plasma membrane protongradient. Key words: Plasma membrane, H⁺ fluxes, ATPase, zinc deficiency     

Release of Membrane-Bound Trails by Trypanosoma cruzi Amastigotes onto Modified Surfaces and Mammalian Cells

ABSTRACT. Upon incubation at 37° C onto glass coverslips coated with Concanavalin A, poly-L-lysine, or a monoclonal antibody (1D9) directed to the parasite major surface glycoprotein Ssp-4, extracellular Trypanosoma cruzi amastigotes release trails of material barely visible by light microscopy. This release is not associated with parasite movements. Immunolabeling studies confirmed that the material is derived from the parasite's membrane since thin section through samples labeled with 1D9 revealed that the trails are membrane-bound structures. Scanning electron microscopy showed that the ∼0.1-μm thick trails of material emerging from the amastigotes can be uniform or beaded, indicating a tendency to vesiculation. The trails are preferentially released from the flagellar pocket region and/or at the opposite posterior end of the parasite body, and seem to be devoid of microtubules. The release is time and temperature-dependent and fixed parasites do not form trails. All attempts to inhibit trail release using drugs (antimycin A, sodium azide, cytochalasin D, nocodazole, genistein, staurosporine, EGTA) failed. The observation of trails associated with intracellular parasites and amastigotes invading Vero cells suggests that this is probably a physiological process.     

Competition and resistance to starvation in larvae of container-inhabiting Aedes mosquitoes

Abstract. 1. Hypotheses about declining populations of container-inhabiting Aedes mosquitoes following the invasion by additional species were tested.
2. The larval competition hypothesis was studied experimentally in pure and mixed cultures of Aedes aegypti (L.), A.albopictus (Skuse) and A.triseriatus (Say). The experiments used decomposing leaf litter in the laboratory, as opposed to most previous research which used non-natural food.
3. Resistance to starvation is introduced as a new measure of larval performance and competitiveness. The hypothesis is that more successful larvae store larger energy reserves and resist the lack of food longer.
4. Contrary to previous research showing better performance of A.aegypti in mixed cultures, A.albopictus developed faster and had greater survival when natural food was used.
5. Resistance to starvation was greater in the better performing species (i.e. A.aegypti with non-natural food and A.albopictus with leaf litter). Oxygen consumption by starved larvae was similar in the three container species, and in the ground-water mosquito, A.taeniorhynchus (Wied.), whose resistance to starvation was comparatively very low.     

Inhibition of Protein Synthesis and Amino Acid Transport by Crystal Violet in Trypanosoma cruzi

ABSTRACT. [³⁵S]methionine incorporation into proteins of either T. cruzi epimastigotes or trypomastigotes was drastically inhibited by low concentrations of crystal violet in a dose-dependent manner. This inhibition was not due to ATP depletion since cellular ATP levels did not change significantly after incubation of epimastigotes with 50 μM crystal violet for similar periods of time, and was unaffected by changes in the extracellular free calcium concentration. Although crystal violet was able to inhibit protein synthesis in a cell-free system from T. cruzi epimastigotes, half maximal inhibition was at 1 mM, a concentration three orders of magnitude higher than those that inhibited protein synthesis in intact cells. On the other hand, crystal violet was able to inhibit total [³⁵S]methionine uptake at similar concentrations to those that inhibited protein synthesis while addition of increasing concentrations of cold methionine to the incubation medium protected the cells against crystal violet inhibition. Crystal violet also inhibited total [³H]proline uptake thus indicating that it has a general inhibitory effect upon the transport of amino acids, and not specifically upon methionine. These results indicate that inhibition of protein synthesis by crystal violet is probably due to inhibition of amino acid uptake.     

The extant Chaoborus assemblage can be assessed using subfossil mandibles

1. Assemblages of Chaoborus were examined in 80 thermally stratified southern central Canadian Shield lakes to explore whether subfossil mandibles could be useful in assemblage‐level studies of Chaoborus. 2. Chaoborus (Sayomyia) (probably Chaoborus punctipennis in this study region) and Chaoborus flavicans were the most common taxa recorded, while Chaoborus trivittatus was rarer. Chaoborus americanus was not recorded in subfossil assemblages, because no fishless lakes were included in this study. Chaoborus flavicans had higher relative abundances (%) in lakes with higher dissolved organic carbon (DOC), probably because of reduced fish predation in less transparent water. 3. Results from logistic regression indicate that patterns of presence/absence for Chaoborus mandibles in the study lakes were influenced primarily by hypolimnetic oxygen concentration, probably because of the presence or extent of a hypolimnetic refugium from fish predation. 4. Chaoborus species richness in lakes, derived from subfossil assemblages, did not differ significantly from species richness estimates derived from plankton sampling with a net. Patterns of dominance and coexistence [e.g. the widespread co‐occurrence of C. flavicans and C. (Sayomyia)] determined from subfossil assemblages agreed with previous studies of the contemporary living assemblage. 5. These results suggest that subfossil assemblages may be used as an alternative to nocturnal plankton sampling to carry out research on the community ecology of Chaoborus. 6. We propose a hierarchical conceptual model of assemblage‐level patterns of Chaoborus in temperate lakes. Chaoborus americanus dominates in fishless lakes, whereas in lakes with fish Chaoborus is typically absent where there is no anoxic hyplimnion. In lakes with anoxic strata, C. trivittatus tends to dominate in lakes with few fish; in the remaining lakes, C. flavicans and C. (Sayomyia) dominate, although C. flavicans is more relatively abundant in lakes with lower water clarity (higher DOC).     

NO signalling in cytokinin-induced programmed cell death

Cell death can be induced by cytokinin 6-benzylaminopurine (BA) at high dosage in suspension-cultured Arabidopsis cells. Herein, we provide evidence that BA induces nitric oxide (NO) synthesis in a dose-dependent manner. A reduction in cell death can be observed when the cytokinin is supplemented with the NO scavenger 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (cPTIO) or the nitric oxide synthase (NOS) inhibitors: 2-aminoethyl-isothiourea (AET) and N^G.-monomethyl- l -arginine ( l -NMMA), which suggests that NO is produced via a NOS and is a signalling component of this form of programmed cell death. In BA-treated cells, mitochondrial functionality is altered via inhibition of respiration. This inhibition can be prevented by addition of either cPTIO or AET implying that NO acts at the mitochondrial level.     

Functional bases of host-acceptance behaviour in the aphid parasitoid Aphidius ervi

Abstract.  The host acceptance behaviour in Aphidius ervi is investigated, assessing the role of both external and internal host-associated cues, offered to the experimental parasitoids with parafilm-made aphid dummies. The reaction to internal cues present in the host haemolymph is clearly evident, and its intensity is enhanced by external cues. Parasitoid females lay few eggs in aphid dummies filled with host haemolymph. A significant increase in the number of both oviposition reactions (host stinging) and egg laying is observed only when these dummies are coated with cornicle secretion. However, this enhancement is not observed when the aphid dummies contain distilled water. Thus, the host acceptance behaviour of A. ervi females appears to be controlled by the integration of both external and internal chemical cues. The physiological basis of this behavioural response is investigated with a detailed study on the anatomy and ultrastructure of A. ervi ovipositor. The detection of chemical cues present in the host haemolymph that act as kairomones is made possible by the presence of gustatory sensilla on the tip of the ovipositor. These sensilla consist of porous areas, reached by unbranched dendrites running inside both the lower valves (i.e. first valvulae) and the unpaired upper valve (i.e. second valvulae). The mechanosensory function during oviposition appears to be provided mostly by the basiconic sensilla found on the tip of external valves (i.e. third valvulae). A tentative functional model accounting for the observed oviposition behaviour of A. ervi is proposed.