4,792 research outputs found

    Bloodstream form trypanosoma brucei depend upon multiple metacaspases associated with RAB11-positive endosomes

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    Trypanosoma brucei possesses five metacaspase genes. Of these, MCA2 and MCA3 are expressed only in the mammalian bloodstream form of the parasite, whereas MCA5 is expressed also in the insect procyclic form. Triple RNAi analysis showed MCA2, MCA3 and MCA5 to be essential in the bloodstream form, with parasites accumulating pre-cytokinesis. Nevertheless, triple null mutants (Δmca2/3Δmca5) could be isolated after sequential gene deletion. Thereafter, Δmca2/3Δmca5 mutants were found to grow well both in vitro in culture and in vivo in mice. We hypothesise that metacaspases are essential for bloodstream form parasites, but they have overlapping functions and their progressive loss can be compensated for by activation of alternative biochemical pathways. Analysis of Δmca2/3Δmca5 revealed no greater or lesser susceptibility to stresses reported to initiate programmed cell death, such as treatment with prostaglandin D2. The metacaspases were found to colocalise with RAB11, a marker for recycling endosomes. However, variant surface glycoprotein (VSG) recycling processes and the degradation of internalised anti-VSG antibody were found to occur similarly in wild type, Δmca2/3Δmca5 and triple RNAi induced parasites. Thus, the data provide no support for the direct involvement of T. brucei metacaspases in programmed cell death and suggest that the proteins have a function associated with RAB11 vesicles that is independent of known recycling processes of RAB11-positive endosomes

    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

    Stiffness and energy losses in cylindrically symmetric superconductor levitating systems

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    Stiffness and hysteretic energy losses are calculated for a magnetically levitating system composed of a type-II superconductor and a permanent magnet when a small vibration is produced in the system. We consider a cylindrically symmetric configuration with only vertical movements and calculate the current profiles under the assumption of the critical state model. The calculations, based on magnetic energy minimization, take into account the demagnetization fields inside the superconductor and the actual shape of the applied field. The dependence of stiffness and hysteretic energy losses upon the different important parameters of the system such as the superconductor aspect ratio, the relative size of the superconductor-permanent magnet, and the critical current of the superconductor are all systematically studied. Finally, in view of the results, we provide some trends on how a system such as the one studied here could be designed in order to optimize both the stiffness and the hysteretic losses.Comment: 8 pages; 8 figure

    Intergenerational equity in municipal accounting: New Zealand 1910s

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    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

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

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    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)

    Ecology of common bully (Gobiomorphus cotidianus) in the Tarawera and Rangitaiki rivers: isolation by inland distance or anthropogenic discharge?

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    Previous research has identified distinct genetic, life-history and reproductive differences between populations of common bully (Gobiomorphus cotidianus) upstream and downstream of a pulp and paper mill outfall on the Tarawera River in the Bay of Plenty, New Zealand. This study investigated the distribution of common bully in the Tarawera River by examining fish collected from upstream (37 km inland) and downstream (20 km inland) locations and comparing them to fish from similar inland locations (40 km and 17 km inland, respectively) in the nearby Rangitaiki River. Reproductive divergence was observed between upstream and downstream sites of both rivers by differing annual trends in gonadosomatic index. Stable carbon and nitrogen isotopes confirmed residency at each sampling site and otolith microchemistry demonstrated different life-history strategies between upstream and downstream populations. Diadromous recruits dominated in both downstream river populations, with a general disappearance of diadromy upstream. A mixture of diadromous and non-diadromous fish were found in the upstream Rangitaiki River, whereas diadromous recruits were absent in the upstream Tarawera River. A reduction in oculoscapular canal structures also coincided with loss of diadromy in fish from both rivers. A behavioural study to determine whether pulp and paper mill effluent may deter fish migration within the Tarawera River demonstrated a strong avoidance of effluent, but only at concentrations (>25%) greater than those that naturally occur in the river (<15%). The results of this study suggest that combinations of influences coupled with inland distance are likely to be responsible for the isolation of common bully subpopulations within the Tarawera River

    The influence of defined ante-mortem stressors on the early post-mortem biochemical processes in the abdominal muscle of the Norway lobster, Nephrops norvegicus (Linnaeus, 1758)

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    The effects of four different ante-mortem stressors (exercise, emersion, starvation and a patent infection with the parasite Hematodinium sp.) on post-mortem processes have been investigated in the abdominal muscle of Norway lobster Nephrops norvegicus by measuring changes in the pH, the levels of glycogen, l-lactate, arginine phosphate, ATP, ADP, AMP, IMP, HxR, Hx and the adenylate energy charge (AEC) over a time course of 24 h with samples being taken at 0, 3, 6, 12 and 24 h. The acute stresses of intense exercise and 2 h emersion resulted in a premature onset of anaerobic glycolysis, leading both to an enhanced glycogen depletion rate and an early accumulation of l-lactate. The chronic stressors, starvation and parasite infection, resulted in a complete ante-mortem depletion of muscle glycogen and consequently the failure of post-mortem glycolytic fermentation. Post-mortem pH and ATP inter-conversion were significantly altered in chronically stressed animals. Ante-mortem, a rapid, almost complete depletion of arginine phosphate was observed in all stress groups. The AEC was altered significantly by all stresses, indicating a strong energy demand. The findings suggest that ante-mortem stressors strongly influence the post-mortem biochemical processes. The laboratory-based results are compared to 'field' data and effects on post-harvest product quality are discussed
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