Biochemical Requirements for Intracellular Invasion by Trypanosoma cruzi: Protein Synthesis1

The effects of irreversible inhibition of protein synthesis by pactamycin in either infective forms of Trypanosoma cruzi or mammalian host cells on cellular invasion by this human pathogen were investigated. Treatment of bloodstream forms of T. cruzi with pactamycin markedly reduced their ability to bind either fibroblast-like cells of monkey origin or myoblasts of rat origin. The number of amastigote forms that could be established intracellularly was also significantly decreased with respect to control values obtained when mock-treated (medium alone) trypomastigotes were incubated with the cells. Pactamycin treatment also reduced the infectivity of T. cruzi trypomastigotes for mice as evidenced by both significantly reduced parasitemia levels and mortality rates when compared with those of control mice infected with mock-treated parasites. Inhibition of protein synthesis in the host cells neither prevented cell infection by untreated trypomastigotes nor altered the percentages of infected cells or the magnitude of the infection in vitro. These results indicate that protein synthesis is a requirement for cell invasion by T. cruzi and that the parasite can establish itself and replicate within cells relying on its own protein synthesis ability.     

Ultrastructural Changes in Cotyledons of Pineapple Guava (Myrtaceae) During Somatic Embryogenesis

Ultrastructural studies (SEM and TEM) were performed on cotyledonsof pineapple guava ( Feijoa sellowiana Berg, Myrtaceae) inducedto form embryos on medium containing 1.0 mg l^-1(4.5µM2,4-D) and 0.3M sucrose. At the time of culture, the cells werefilled with protein and lipid bodies. Microbodies and poorlydifferentiated organelles could also be seen. In contrast togerminating cotyledons, where lipid and protein reserves werequickly metabolized, cells of the embryogenically induced cotyledonsshowed evidence of reserve consumption only after 5 d of culture.Subepidermal cells of the upper cotyledonary surface underwentseveral divisions giving rise to a meristematic layer of severalcells thickness from which somatic embryos developed. Embryoscould also be formed directly by successive divisions of epidermalcells. Cells involved in somatic embryo formation containeda large nucleus with a conspicuous nucleolus and dense cytoplasmwhere numerous ribosomes, mitochondria, plastids with starchand short profiles of rough endoplasmic reticulum were present.Plasmodesmata were present both in cell walls of the meristematiccells and in few celled embryos whereas in degenerating embryosor in more advanced stages of somatic embryo development noplasmodesmata could be found. Although oil bodies were not observedin the meristematic cells they were identified in very youngembryos, being the first reserve compounds to appear. Cellsnot involved in somatic embryo differentiation were characterizedby the presence of several microbodies containing a crystalloidinclusion and elongated mitochondria. Feijoa sellowiana ; pineapple guava; somatic embryogenesis; ultrastructural studies     

Assimilatory Sulfate Reduction by the Hemoflagellate Leishmania tarentolae1

SYNOPSIS. Continuous growth of one cell line (UCI variant) of Leishmania tarentolae was achieved in the absence of organic sulfur. These cells were able to use sodium sulfate, and, to a limited extent, sodium sulfite as their sole sulfur source and could utilize methionine sulfoxide in place of L-methionine. A related cell line (RU variant) was unable to grow in organic sulfate-free media nor could these cells utilize methionine sulfoxide. UCI promastigotes incorporated significant amounts of ³⁵S sodium sulfate; killed cells did not take up the label. ³⁵S incorporation was inhibited by sodium molybdate (5 × 10^?4 M), sodium arsenite (5 × 10^?4 M), 2,4-dinitrophenol (1 × 10^?4 M), or KCN (5 × 10^?4 M). RU promastigotes did not incorporate significant amounts of ³⁵S sodium sulfate. Thin layer chromatographs of protein hydrolysates from UCI cells incubated in ³⁵S sodium sulfate revealed several radio opaque spots, one of which had chromatographic properties of cystine. UCI variants of L. tarentolae were therefore capable of assimilatory sulfate reduction whereas RU cells lacked this ability.     

Use of Camera-Trapping to Estimate Puma Density and Influencing Factors in Central Brazil

Abstract: We used remotely triggered cameras to collect data on Puma (Puma concolor) abundance and occupancy in an area of tropical forest in Brazil where the species' status is poorly known. To evaluate factors influencing puma occupancy we used data from 5 sampling campaigns in 3 consecutive years (2005 to 2007) and 2 seasons (wet and dry), at a state park and a private forest reserve. We estimated puma numbers and density for the 2007 sampling data by developing a standardized individual identification method. We based individual identification on 1) time-stable parameters (SP; physical features that do not change over time), and 2) time-variable parameters (VP; marks that could change over time such as scars and botfly marks). Following individual identification we established a capture-recapture history and analyzed it using closed population capture-mark-recapture models. Puma capture probability was influenced by camera placement (roads vs. trails), sampling year, and prey richness. Puma occupancy was positively associated with species richness and there was a correlation between relative puma and jaguar (Panthera onca) abundance. Identifications enabled us to generate 8 VP histories for each photographed flank, corresponding to 8 individuals. We estimated the sampled population at 9 pumas (SE = 1.03, 95% CI = 8–10 individuals) translating to a density of 3.40 pumas/100 km². Information collected using camera-traps can effectively be used to assess puma population size in tropical forests. As habitat progressively disappears and South American felines become more vulnerable, our results support the critical importance of private forest reserves for conservation.