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During development in the mosquito midgut, malarial parasites must traverse a chitin-containing peritrophic matrix (PM) that forms around the food bolus. Previously Huber et al. [Huber, M., Cabib, E. & Miller, L. H. (1991) Proc. Natl. Acad. Sci. USA 88, 2807USA 88, -2810 reported that the parasite secretes a protein with chitinase activity, and they suggested that parasite chitinase (EC 3.2.1.14) plays an important role in the parasite's egress from the blood meal. We found that allosamidin, a specific inhibitor of chitinase, completely blocked oocyst development in vivo and thus blocked malaria parasite transmission. Addition of exogenous chitinase to the blood meal prevented the PM from forming and reversed the transmission-blocking activity of allosamidin. Using exogenous chitinase, we also found that the PM does not limit the number of parasites that develop into oocysts, suggesting that the parasite produces sufficient quantities of chitinase to penetrate this potential barrier. In addition, we found that treatment of parasite chitinase with a diisopropyl fluorophosphatesensitive trypsinlike protease from the mosquito midgut or endoproteinase Lys-C increased its enzymatic activity. These results suggest that malaria parasite has evolved an intricate mechanism to adapt to the PM and the protease-rich environment of the mosquito midgut.

The role of the peritrophic matrix (PM) in the development of Leishmania major infections in a natural vector, Phlebotomus papatasi, was investigated by addition of exogenous chitinase to the bloodmeal, which completely blocked PM formation. Surprisingly, the absence of the PM was associated with the loss of midgut infections. The chitinase was not directly toxic to the parasite, nor were midgut infections lost due to premature expulsion of the bloodmeal. Most parasites were killed in chitinase-treated flies within the first 4 h after feeding. Substantial early killing was also observed in control flies, suggesting that the lack of PM exacerbates lethal conditions which normally exist in the blood-fed midgut. Early parasite mortality was reversed by soybean trypsin inhibitor. Allosamadin, a specific inhibitor of chitinase, led to a thickening of the PM, and also prevented the early parasite mortality seen in infected flies. Susceptibility to gut proteases was extremely high in transitional-stage parasites, while amastigotes and fully transformed promastigotes were relatively resistant. A novel role for the PM in promoting parasite survival is suggested, in which the PM creates a barrier to the rapid diffusion of digestive enzymes, and limits the exposure of parasites to these enzymes during the time when they are especially vulnerable to proteolytic damage.

Nanoporous carbon materials are a versatile source of carbons that would be useful in applications ranging from electronics to electrochemical energy storage. Here, we focus on nanoporous carbon materials prepared by direct carbonization of zeolitic imidazolate frameworks (ZIF-8) towards supercapacitor applications. Several types of nanoporous carbons have been prepared by varying the applied carbonization temperature. The symmetric devices assembled using nanoporous carbon electrodes were tested for their optimal performance in the electrolyte of sulfuric acid solution. We demonstrate the effects of various factors (e.g., surface area, nitrogen content, degree of graphitization, and relative percentage of micropores) on the performance.

No abstract

Malaria is a devastating public health menace, killing over one million people every year and infecting about half a billion. Here it is shown that the protozoan Plasmodium gallinaceum, a close relative of the human malaria parasite Plasmodium falciparum, can develop in the fruit fly Drosophila melanogaster. Plasmodium gallinaceum ookinetes injected into the fly developed into sporozoites infectious to the vertebrate host with similar kinetics as seen in the mosquito host Aedes aegypti. In the fly, a component of the insect's innate immune system, the macrophage, can destroy Plasmodia. These experiments suggest that Drosophila can be used as a surrogate mosquito for defining the genetic pathways involved in both vector competence and part of the parasite sexual cycle.

No abstract

Penetration of the mosquito midgut epithelium is obligatory for the further development of Plasmodium parasites. Therefore, blocking the parasite from invading the midgut wall disrupts the transmission of malaria. Despite such a pivotal role in malaria transmission, the cellular and molecular interactions that occur during the invasion are not understood. Here, we demonstrate that the ookinetes of Plasmodium gallinaceum, which is related closely to the human malaria parasite Plasmodium falciparum, selectively invade a cell type in the Aedes aegypti midgut. These cells, unlike the majority of the cells in the midgut, do not stain with a basophilic dye (toluidine blue) and are less osmiophilic. In addition, they contain minimal endoplasmic reticulum, lack secretory granules, and have few microvilli. Instead, these cells are highly vacuolated and express large amounts of vesicular ATPase. The enzyme is associated with the apical plasma membrane, cytoplasmic vesicles, and tubular extensions of the basal membrane of the invaded cells. The high cost of insecticide use in endemic areas and the emergence of drug resistant malaria parasites call for alternative approaches such as modifying the mosquito to block the transmission of malaria. One of the targets for such modification is the parasite receptor on midgut cells. A step toward the identification of this receptor is the realization that malaria parasites invade a special cell type in the mosquito midgut. This is the centennial of the discovery in 1897 by Ronald Ross that mosquitoes transmit malaria (1). In the 1890s, fertilization of malaria parasites to form zygotes, the first step of Plasmodium development in the mosquito, also was discovered. Zygotes develop into ookinetes within the midgut and penetrate the peritrophic matrix and the midgut epithelium. In the hemolymph, parasites develop as oocysts between the midgut epithelium and basal lamina. Sporozoites develop during the hemolymph stage and penetrate the salivary glands to be injected into the human during blood feeding (2, 3). One of the most crucial aspects of the sporogonic development of malaria parasites is the crossing of the mosquito midgut epithelium by the ookinetes. The ookinetes that fail to penetrate the midgut epithelium die in the gut lumen. Blocking ookinete penetration of mosquito midgut epithelium, therefore, is a potential strategy to block malaria transmission. To exploit this invasion process, however, a better understanding of ookinete interaction with the midgut is necessary. The molecular aspects of this penetration process currently are not known.The ultrastructure of ookinetes within the mosquito midgut epithelium has been studied (4-7). In these experiments, only a few parasites were observed because invasion and transit across the epithelial cells are asynchronous. To examine the invasion process more extensively, we have developed an in vitro invasion assay. This assay, along with an in vivo procedure, has allowed us to examine a large number of invaded cells and has pr...