Inhibition of Early Endosome Fusion by Trypansom cruzi-Infected Macrophage Cytosol

ABSTRACT. Trypanosoma cruzi trypomastigotes survive inside macrophages by promoting fusion between the parasitophorous vacuole and mature host lysosomes upon internalization. Since trypomastigotes can evade the lytic pathway, the earliest steps of endocytosis, such as early endosome fusion, may be affected. To test this hypothesis, we used an in vitro early endosome fusion assay. Our results show that trypomastigote-infected macrophage cytosols cannot promote fusion between early endosomes, compared to mock-infected cytosols (heat-killed trypomastigotes were used in the parasite-macrophage interaction assay). GTPγS addition potentiates the fusogenic activity driven by trypomastigote-infected macrophage cytosol-mediated assays, unlike the biphasic fusogenic effect obtained with GTPγS treatment of macrophage cytosol controls. Calcium-stimulated early endosome fusogenic processes are not affected in the assays mediated by infected macrophage cytosol. We conclude that GTP-regulated factors, and not calcium-regulated elements, are involved in the inhibition of the early endosome fusogenic process by the trypomastigote-infected macrophage cytosol. This primary impediment to the progress of a normal endocytosis may be a relevant step required for the lysosomal recruitment-fusion of the host lysosmes upon trypomastigote infection and further survival of the parasite within its host.     

Seasonal dynamics of the brown dog tick,Rhipicephalus sanguineus,on a confined dog population in Italy

This study evaluated the seasonal dynamics of Rhipicephalus sanguineus (Latreille) (Acari: Ixodidae) on naturally infested dogs in a private shelter in southern Italy. From March to May 2008, 39 autochthonous mixed‐breed young dogs and 10 beagles were enrolled in the study. From March 2008 until March 2009, every 21 ± 2 days, 11 body sites of each dog were checked for ticks. At each follow‐up, the number of ticks, their developmental stage, sex and location on the dog's body were recorded. Adult ticks were found throughout the year, but immatures were absent in January and February. The adult tick population increased from July to August, whereas the load of immatures increased in early July and peaked in September, which suggests that R. sanguineus develops one generation per year in this area. The mean number of immature ticks per infested dog was higher than that of adults from March to October 2008. Ears, interdigital areas and armpits were the most frequent attachment sites of adult ticks. At the last follow‐up, a total of 2266 ticks were collected and identified as R. sanguineus. The results suggest that R. sanguineus develops one generation per year in the study area, but that it infests dogs in all seasons. This information should be taken into account when planning control programmes against this tick species and the pathogens it transmits.     

Molecular systematics of the butterfly tribe Preponini (Nymphalidae: Charaxinae)

The nymphalid butterfly tribe Preponini includes some of the Neotropical region's most spectacular and familiar butterflies, but the taxonomy of the group nevertheless remains unstable. Several recent studies of Nymphalidae phylogeny have suggested that both the tribe itself and several genera might not be monophyletic, but to date taxon sampling has not been sufficiently comprehensive to allow informed revision of the group's systematics. We therefore conducted the first complete species‐level phylogenetic study of the tribe to establish a firm higher classification. We used DNA sequence data from three genes, the two mitochondrial genes cytochrome oxidase subunits I and II (COI and COII), and the nuclear gene elongation factor‐1α (EF‐1α), to reconstruct the phylogeny of the tribe using maximum likelihood (ML), maximum parsimony (MP) and Bayesian inference (BI). We included 48 individuals representing the 22 recognised Preponini species, and an additional 25 out‐group taxa to explore taxonomic limits at different levels. Firstly, we found that Anaeomorpha splendida Rothschild never grouped with remaining Preponini, so that maintaining monophyly of the tribe requires the taxon to be excluded, and we thus reinstate the tribe Anaeomorphini stat.rev. Secondly, we investigated generic limits, in particular the relationship of Noreppa Rydon to Archaeoprepona Fruhstorfer, and that of Agrias Doubleday to Prepona Boisduval. The molecular results coupled with previous morphological studies suggest that Noreppa syn.n should be synonymised with Archaeoprepona, and that Agrias syn.n should be synonymised with Prepona. We found Prepona pheridamas (Cramer) to be sister to all other Prepona, and markedly divergent from them in both morphology and DNA sequences, suggesting the possibility that it should be placed in a separate genus. We also found a number of cases of significant DNA sequence divergence and paraphyly or polyphyly within putative species that require further taxonomic attention, including Prepona claudina (Godart) stat.n. and Prepona narcissus (Staudinger) stat.n., Prepona pylene Hewitson and Prepona deiphile (Godart). Future research should focus on a broader population sampling of widespread, polymorphic Preponini species to thoroughly revise the current species‐level taxonomy, thus creating a solid foundation for studies in ecology and conservation.     

Microtubular Systems of Paramecium in Division: Pattern of Cytospindle Assembly

The pattern of cytospindle assembly and the modifications of the microtubular cytoplasmic network during division of Paramecium are studied by means of indirect immunofluorescence. The assembly of cytospindle starts at two independent areas placed respectively around the proter's and opisthe's buccal overture. The moment of the microtubule bundles’appearance depends on their distance from the buccal opening, with those closest appearing 1st. The existence of microtubule organizing centers that act transiently during division of Paramecium is discussed.     

Sudden expansion of a single brown bear maternal lineage across northern continental Eurasia after the last ice age: a general demographic model for mammals?

The brown bear has proved a useful model for studying Late Quaternary mammalian phylogeography. However, information is lacking from northern continental Eurasia, which constitutes a large part of the species' current distribution. We analysed mitochondrial DNA sequences (totalling 1943 bp) from 205 bears from northeast Europe and Russia in order to characterize the maternal phylogeography of bears in this region. We also estimated the formation times of the sampled brown bear lineages and those of its extinct relative, the cave bear.
Four closely related haplogroups belonging to a single mitochondrial subclade were identified in northern continental Eurasia. Several haplotypes were found throughout the whole study area, while one haplogroup was restricted to Kamchatka. The haplotype network, estimated divergence times and various statistical tests indicated that bears in northern continental Eurasia recently underwent a sudden expansion, preceded by a severe bottleneck. This brown bear population was therefore most likely founded by a small number of bears that were restricted to a single refuge area during the last glacial maximum. This pattern has been described previously for other mammal species and as such may represent one general model for the phylogeography of Eurasian mammals. Bayesian divergence time estimates are presented for different brown and cave bear clades. Moreover, our results demonstrate the extent of substitution rate variation occurring throughout the phylogenetic tree, highlighting the need for appropriate calibration when estimating divergence times.     

cpFBPaseII, a novel redox-independent chloroplastic isoform of fructose-1,6-bisphosphatase

A full-length FBPase cDNA has been isolated from Fragaria  ×  ananassa (strawberry) corresponding to a novel putative chloroplastic FBPase but lacking the regulatory redox domain, a characteristic of the plastidial isoenzyme (cpFBPaseI). Another outstanding feature of this novel isoform, called cpFBPaseII, is the absence of the canonical active site. Enzymatic assays with cpFBPaseII evidenced clear Mg²⁺-dependent FBPase activity and a K _m for fructose-1,6-bisphosphate (FBP) of 1.3 m m . Immunolocalization experiments and chloroplast isolation confirmed that the new isoenzyme is located in the stroma. Nevertheless, unlike cpFBPaseI, which is redox activated, cpFBPaseII did not increase its activity in the presence of either DTT or thioredoxin f (TRX f ) and is resistant to H₂O₂ inactivation. Additionally, the novel isoform was able to complement the growth deficiency of the yeast FBP1 deletion fed with a non-fermentable carbon source. Furthermore, orthologues are restricted to land plants, suggesting that cpFBPaseII is a novel and an intriguing chloroplastic FBPase that emerged late in the evolution of photosynthetic organisms, possibly because of a pressing need of land plants.