Tracking autophagy during proliferation and differentiation of trypanosoma brucei

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 *

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

The Concepts of Reliability and Homogeneity

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.

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

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

Microbiological influences on fracture surfaces of intact mudstone and the implications for geological disposal of radioactive waste

The significance of the potential impacts of microbial activity on the transport properties of host rocks for geological repositories is an area of active research. Most recent work has focused on granitic environments. This paper describes pilot studies investigating changes in transport properties that are produced by microbial activity in sedimentary rock environments in northern Japan. For the first time, these short experiments (39 days maximum) have shown that the denitrifying bacteria, Pseudomonas denitrificans, can survive and thrive when injected into flow-through column experiments containing fractured diatomaceous mudstone and synthetic groundwater under pressurized conditions. Although there were few significant changes in the fluid chemistry, changes in the permeability of the biotic column, which can be explained by the observed biofilm formation, were quantitatively monitored. These same methodologies could also be adapted to obtain information from cores originating from a variety of geological environments including oil reservoirs, aquifers and toxic waste disposal sites to provide an understanding of the impact of microbial activity on the transport of a range of solutes, such as groundwater contaminants and gases (e.g. injected carbon dioxide)

Intergenerational equity in municipal accounting: New Zealand 1910s

Accounting for fixed assets by municipalities has been discussed in the accounting history literature previously. This paper addresses two issues related to accounting for fixed assets not previously discussed; the influence of the principle of intergenerational equity on local government accounting, and the influence of users of accounting information in accounting policy making in government accounting. The paper identifies that users of accounting information have had significant influence in a debate on government accounting policy, and that the principle of intergenerational equity was given a position of high importance in the debate, but not an unchallengeable position. The motivation of the users of accounting information to engage in the debates is identified as a form of civic duty, which is consistent with the ethical imperative in ensuring intergenerational equity

Scanning Tunneling Spectroscopy in MgB2

We present scanning tunneling microscopy measurements of the surface of superconducting MgB2 with a critical temperature of 39K. In zero magnetic field the conductance spectra can be analyzed in terms of the standard BCS theory with a smearing parameter Gamma. The value of the superconducting gap is 5.2 meV at 4.2 K, with no experimentally significant variation across the surface of the sample. The temperature dependence of the gap follows the BCS form, fully consistent with phonon-mediated superconductivity in this novel superconductor. The application of a magnetic field induces strong pair-breaking as seen in the conductance spectra in fields up to 6 T.Comment: 4 pages, 4 figure

The Reliability of Test Discriminations

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/66658/2/10.1177_001316445501500404.pd