5,204 research outputs found

    Tracking autophagy during proliferation and differentiation of trypanosoma brucei

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    Autophagy is a lysosome-dependent degradation mechanism that sequesters target cargo into autophagosomal vesicles. The Trypanosoma brucei genome contains apparent orthologues of several autophagy-related proteins including an ATG8 family. These ubiquitin-like proteins are required for autophagosome membrane formation, but our studies show that ATG8.3 is atypical. To investigate the function of other ATG proteins, RNAi compatible T. brucei were modified to function as autophagy reporter lines by expressing only either YFP-ATG8.1 or YFP-ATG8.2. In the insect procyclic lifecycle stage, independent RNAi down-regulation of ATG3 or ATG7 generated autophagy-defective mutants and confirmed a pro-survival role for autophagy in the procyclic form nutrient starvation response. Similarly, RNAi depletion of ATG5 or ATG7 in the bloodstream form disrupted autophagy, but did not impede proliferation. Further characterisation showed bloodstream form autophagy mutants retain the capacity to undergo the complex cellular remodelling that occurs during differentiation to the procyclic form and are equally susceptible to dihydroxyacetone-induced cell death as wild type parasites, not supporting a role for autophagy in this cell death mechanism. The RNAi reporter system developed, which also identified TOR1 as a negative regulator controlling YFP-ATG8.2 but not YFP-ATG8.1 autophagosome formation, will enable further targeted analysis of the mechanisms and function of autophagy in the medically relevant bloodstream form of T. brucei

    SOME CHARACTERISTICS OF CHOICE BEHAVIOR IN RISKY SITUATIONS *

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    Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/73591/1/j.1749-6632.1961.tb20178.x.pd

    The Concepts of Reliability and Homogeneity

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    Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/67956/2/10.1177_001316445001000103.pd

    Fabric anisotropy & DEM informed two-surface hyperplasticity : constitutive formulation, asymptotic states & experimental validation.

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    In geotechnical analysis continuum idealisations of the bulk material still provide the most appropriate approach for engineers designing large-scale structures. In this area, the most successful framework for describing the behaviour of soils is Critical State (CS) soil mechanics. However, the findings from discrete element method (DEM) analysis, such as the uniqueness of the CS, can provide invaluable information in the development such models. This paper details the key concepts behind a two-surface hyperplasticity model (?) whose development was informed by recent DEM findings on the uniqueness of the CS. Asymptotic states of the model will be confirmed and the DEM-continuum-experimental loop will be closed through comparison of the developed model with experimental data on coarse-grained particulate media. This will demonstrate, that providing the previous stress history is accounted for, the proposed model is suitable for a variety of particulate media

    Metabolomic profiling and stable isotope labelling of Trichomonas vaginalis and Tritrichomonas foetus reveal major differences in amino acid metabolism including the production of 2-hydroxyisocaproic acid, cystathionine and S-methylcysteine

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    Trichomonas vaginalis and Tritrichomonas foetus are pathogens that parasitise, respectively, human and bovine urogenital tracts causing disease. Using LC-MS, reference metabolomic profiles were obtained for both species and stable isotope labelling with D-[U-13C6] glucose was used to analyse central carbon metabolism. This facilitated a comparison of the metabolic pathways of T. vaginalis and T. foetus, extending earlier targeted biochemical studies. 43 metabolites, whose identities were confirmed by comparison of their retention times with authentic standards, occurred at more than 3-fold difference in peak intensity between T. vaginalis and T. foetus. 18 metabolites that were removed from or released into the medium during growth also showed more than 3-fold difference between the species. Major differences were observed in cysteine and methionine metabolism in which homocysteine, produced as a bi-product of trans-methylation, is catabolised by methionine γ-lyase in T. vaginalis but converted to cystathionine in T. foetus. Both species synthesise methylthioadenosine by an unusual mechanism, but it is not used as a substrate for methionine recycling. T. vaginalis also produces and exports high levels of S-methylcysteine, whereas only negligible levels were found in T. foetus which maintains significantly higher intracellular levels of cysteine. 13C-labeling confirmed that both cysteine and S-methylcysteine are synthesised by T. vaginalis; S-methylcysteine can be generated by recombinant T. vaginalis cysteine synthase using phosphoserine and methanethiol. T. foetus contained higher levels of ornithine and citrulline than T. vaginalis and exported increased levels of putrescine, suggesting greater flux through the arginine dihydrolase pathway. T. vaginalis produced and exported hydroxy acid derivatives of certain amino acids, particularly 2-hydroxyisocaproic acid derived from leucine, whereas negligible levels of these metabolites occurred in T. foetus

    Preliminary results of spectral reflectance studies of tycho crater

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    The preliminary analysis and interpretation of near infrared spectra obtained for both the interior and exterior deposits associated with the Tycho crater is presented. Specific objectives were: (1) to determine the composition and stratigraphy of the highland crust in the Tycho target site; (2) to determine the likely composition of the primary ejecta which may be present in ray deposits; (3) to investigate the nature of spectral units defined in previous studies; (4) to further investigate the nature and origin of both the bright and dark haloes around the rim crest; and (5) to compare the compositions determined for the Tycho units with those of the Aristarchus crater as well as typical highland deposits. The spectra obtained for the interior areas exhibit similar spectral features. These include relatively strong 1 micron absorption bands whose minima are centered between 0.97 and 0.99 microns and shallow to intermediate continuum slopes. The spectra generally exhibit indications of a 1.3 micron feature consistent with the presence of Fe(2+) bearing plagioclase feldspar. The strong 1 micron absorption features indicate a dominant high Ca clinopyroxene component. Results obtained from the ejecta deposits show that the spectrum of the inner, bright halo is almost identical with those obtained for interior units. The spectrum of the dark halo exhibits a wide, relatively shallow absorption feature centered at 1.01 microns, a 1.3 micron absorption, and a steep continuum slope. This spectrum is interpreted as indicating the presence of pyroxene, Fe-bearing feldspar, and a significant component of Fe-bearing impact melt glass. Finally, the spectra of spots inside Tycho show similarity with certain spectra for Aristarchus. However, the suite of spectra obtained for Tycho exhibits a different trend in terms of band center versus width
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