Transport proteins determine drug sensitivity and resistance in a protozoan parasite, Trypanosoma brucei

Drug resistance in pathogenic protozoa is very often caused by changes to the ‘transportome’ of the parasites. In Trypanosoma brucei, several transporters have been implicated in uptake of the main classes of drugs, diamidines and melaminophenyl arsenicals. The resistance mechanism had been thought to be due to loss of a transporter known to carry both types of agents: the aminopurine transporter P2, encoded by the gene TbAT1. However, although loss of P2 activity is well-documented as the cause of resistance to the veterinary diamidine diminazene aceturate (Berenil®), cross-resistance between the human-use arsenical melarsoprol and the diamidine pentamidine (MPXR) is the result of loss of a separate High Affinity Pentamidine Transporter (HAPT1). A genome-wide RNAi library screen for resistance to pentamidine, published in 2012, gave the key to the genetic identity of HAPT1 by linking the phenomenon to a locus that contains the closely related T. brucei aquaglyceroporin genes TbAQP2 and TbAQP3. Further analysis determined that knockdown of only one pore, TbAQP2, produced the MPXR phenotype. TbAQP2 is an unconventional aquaglyceroporin with unique residues in the “selectivity region” of the pore, and it was found that in several MPXR lab strains the WT gene was either absent or replaced by a chimeric protein, recombined with parts of TbAQP3. Importantly, wild-type AQP2 was also absent in field isolates of T. b. gambiense, correlating with the outcome of melarsoprol treatment. Expression of a wild-type copy of TbAQP2 in even the most resistant strain completely reversed MPXR and re-introduced HAPT1 function and transport kinetics. Expression of TbAQP2 in Leishmania mexicana introduced a pentamidine transport activity indistinguishable from HAPT1. Although TbAQP2 has been shown to function as a classical aquaglyceroporin it is now clear that it is also a high affinity drug transporter, HAPT1. We discuss here a possible structural rationale for this remarkable ability

Research into the views of two child reference groups on the arts in research concerning wellbeing

Introduction: This paper explores an aspect of the process and outcomes of two children’s reference groups: the ways the young members of the reference groups consider the role of the arts in research concerning wellbeing. Methods: Two case studies draw on qualitative data from the projects offering insight into child reference group work over the life of a research project: from recommendations to implementation. Results: The findings demonstrate that both reference groups enabled the research to achieve a fuller engagement with meaning making in relation to data collection by consulting with children. Conclusion: The data reveal how the members offered “insider knowledge” to the researchers concerning the role of the arts in research concerning wellbeing. Themes within the analysis about the impact of the reference group’s knowledge and recommendations on the research project’s data collection methods include the role of the arts connected to empathy, safety and confidentiality and in relation to difference and choice

Implementation of a geometrically informed and energetically constrained mesoscale eddy parameterization in an ocean circulation model

The global stratification and circulation of the ocean and their sensitivities to changes in forcing depend crucially on the representation of the mesoscale eddy field. Here, a geometrically informed and energetically constrained parameterization framework for mesoscale eddies --- termed GEOMETRIC --- is proposed and implemented in three-dimensional primitive equation channel and sector models. The GEOMETRIC framework closes mesoscale eddy fluxes according to the standard Gent--McWilliams scheme, but with the eddy transfer coefficient constrained by the depth-integrated eddy energy field, provided through a prognostic eddy energy budget evolving with the mean state. It is found that coarse resolution calculations employing GEOMETRIC broadly reproduce model sensitivities of the eddy permitting reference calculations in the emergent circumpolar transport, meridional overturning circulation profile and the depth-integrated eddy energy signature; in particular, eddy saturation emerges in the sector configuration. Some differences arise, attributed here to the simple prognostic eddy energy budget employed, to be improved upon in future investigations. The GEOMETRIC framework thus proposes a shift in paradigm, from a focus on how to close for eddy fluxes, to focusing on the representation of eddy energetics.Comment: 19 pages, 9 figures, submitted to Journal of Physical Oceanography; comments welcome. (Copyright statement: see section 7a of https://www.ametsoc.org/ams/index.cfm/publications/ethical-guidelines-and-ams-policies/ams-copyright-policy/

Environmental and Social Influences on the Behaviour of Free-Living Mandarin Ducks in Richmond Park

This is the final version. Available on open access from MDPI via the DOI in this recordData Availability Statement: Data is available upon reasonable request from the corresponding author.Many species of birds are housed in zoos globally and are some of the most popular of animals kept under human care. Careful observations of how species live and behave in their natural habitats can provide us with important knowledge about their needs, adaptations, and internal states, allowing identification of those behaviours that are most important to the individual’s physical health and wellbeing. For this study, Mandarin Ducks (Aix galericulata) were chosen as a study species because, like many species of waterfowl, they are widely kept in both private institutions and zoos, yet little research has been conducted on their core needs in captivity. A free-living population of naturalised Mandarin Ducks living in Richmond Park was used for this research. Data on state behaviours (resting, swimming, foraging, perching, preening, and vigilance) were collected five days a week (08:00–18:00) from the 26 March to 26 May 2021. Secondly, temporal, seasonal, environmental, and animal-centric factors (e.g., Sex) were recorded to assess any impact on the Mandarin’s time-activity budget. Lastly, a comparison between free-living anmd captive activity was conducted (via the literature) to evaluate whether captive behaviours differ to how they are expressed in the wild. Results showed that free-living Mandarins predominantly rested (19.88% ± 28.97), swam (19.57% ± 19.43) and foraged (19.47% ± 25.82), with variations in activity related to factors such as vegetation cover and pond size. Results also showed differences between the time-budgets of free-living and captive Mandarins, suggesting that captive birds may not always have the opportunity to express species-typical behaviours. This research indicated that study of natural behaviours performed in the wild may help to evaluate “normal” behaviour patterns of zoo-housed individuals and provide evidence for environmental and husbandry alterations that can promote good welfare. However, any potential impact on the activity patterns of free-living species due to human interactions should be considered when assessing deviations between the behaviour of wild and captive individual

Eddy saturation and frictional control of the Antarctic Circumpolar Current

The Antarctic Circumpolar Current is the strongest current in the ocean and has a pivotal impact on ocean stratification, heat content, and carbon content. The circumpolar volume transport is relatively insensitive to surface wind forcing in models that resolve turbulent ocean eddies, a process termed “eddy saturation.” Here a simple model is presented that explains the physics of eddy saturation with three ingredients: a momentum budget, a relation between the eddy form stress and eddy energy, and an eddy energy budget. The model explains both the insensitivity of circumpolar volume transport to surface wind stress and the increase of eddy energy with wind stress. The model further predicts that circumpolar transport increases with increased bottom friction, a counterintuitive result that is confirmed in eddy-permitting calculations. These results suggest an unexpected and important impact of eddy energy dissipation, through bottom drag or lee wave generation, on ocean stratification, ocean heat content, and potentially atmospheric CO2

Turbulent drag on a low-frequency vibrating grid in superfluid He-4 at very low temperatures

We present measurements of the dissipative turbulent drag on a vibrating grid in superfluid He-4 over a wide range of (low) frequencies. At high velocities, the dissipative drag is independent of frequency and is approximately the same as that measured in normal liquid He-4. We present measurements on a similar grid in superfluid He-3-B at low temperatures which shows an almost identical turbulent drag coefficient at low frequencies. However, the turbulent drag in He-3-B is substantially higher at higher frequencies. We also present measurements of the inertial drag coefficient for grid turbulence in He-4. The inertial drag coefficient is significantly reduced by turbulence in both superfluid and normal liquid He-4

Fatal encephalitis due to the scuticociliate Uronema nigricans in sea-caged, southern bluefin tuna Thunnus maccoyii

A syndrome characterized by atypical swimming behaviour followed by rapid death was first reported in captive southern bluefin tuna Thunnus maccoyii (Castelnau) in the winter of 1993. The cause of this behaviour was found to be a parasitic encephalitis due to the scuticociliate Uronema nigricans (Mueller). Based on parasitological and histological findings, it is proposed that the parasites initially colonise the olfactory rosettes and then ascend the olfactory nerves to eventually invade the brain. Possible epidemiological factors involved in the pathogenesis of the disease include water temperature (>18 degrees C) and the immune status of the fish

Trypanosoma brucei bloodstream forms express highly specific and separate transporters for adenine and hypoxanthine; evidence for a new protozoan purine transporter family?

The transport of nucleobases and nucleosides in protozoan parasites is known to be performed by Equilibrative Nucleoside Transporter (ENT) family members, including the extensively studied P1 and P2 nucleoside transporters of T. brucei bloodstream forms. Studies with P2 knockout parasites suggested the existence of as yet uncharacterised purine transport mechanisms in these cells. Here, we deleted several ENT genes, in addition to P2, including an array comprising three genes encoding for high-affinity broad-selectivity nucleobase transporters - the longest multi-gene locus deletion in T. brucei to date. It was verified that none of them appreciably contributed to the transport of hypoxanthine in bloodstream forms grown axenically in HMI-9 medium, which was mainly performed by a previously not described hypoxanthine-specific transporter (HXT1) with a Km of 22 ± 1.7 μM and Vmax of 0.49 ± 0.06 pmol(107 cells)-1s-1. The uptake of adenine was also assessed in the knockout cells and was performed by a highly specific adenine transporter (ADET1) with a Km of 573 ± 62 nM and Vmax of 0.23 ± 0.06 pmol(107 cells)-1 s-1. Neither HXT1 nor ADET1 displayed any affinity for other natural purines or pyrimidines and could not be completely inhibited by hypoxanthine or adenine analogues. These carriers may be the final pieces in the substantial transporter array trypanosomes can employ to fine-tune the uptake of purines from diverse environments during their life cycles, and may be encoded by genes other than those of the ENT family

Differing Mechanisms Underlie Sexual Size-Dimorphism in Two Populations of a Sex-Changing Fish

Variability in the density of groups within a patchy environment lead to differences in interaction rates, growth dynamics and social organization. In protogynous hermaphrodites there are hypothesised trade-offs among sex-specific growth, reproductive output and mortality. When differences in density lead to changes to social organization the link between growth and the timing of sex-change is predicted to change. The present study explores this prediction by comparing the social organisation and sex-specific growth of two populations of a protogynous tropical wrasse, Halichoeres miniatus, which differ in density. At a low density population a strict harem structure was found, where males maintained a tight monopoly of access and spawning rights to females. In contrast, at a high density population a loosely organised system prevailed, where females could move throughout multiple male territories. Otolith microstructure revealed the species to be annual and deposit an otolith check associated with sex-change. Growth trajectories suggested that individuals that later became males in both populations underwent a growth acceleration at sex-change. Moreover, in the high density population, individuals that later became males were those individuals that had the largest otolith size at hatching and consistently deposited larger increments throughout early larval, juvenile and female life. This study demonstrates that previous growth history and growth rate changes associated with sex change can be responsible for the sexual dimorphism typically found in sex-changing species, and that the relative importance of these may be socially constrained

Gill's model of the Antarctic Circumpolar Current, revisited: the role of latitudinal variations in wind stress

Adrian Gill’s (1968) model of the Antarctic Circumpolar Current (ACC) is reinterpreted for a stratified, reduced-gravity ocean, where the barotropic streamfunction is replaced by the pycnocline depth, and the bottom drag coefficient by the Gent and McWilliams eddy diffusivity. The resultant model gives a simple description of the lateral structure of the ACC that is consistent with contemporary descriptions of ACC dynamics. The model is used to investigate and interpret the sensitivity of the ACC to the latitudinal profile of the surface wind stress. A substantial ACC remains when the wind jet is shifted north of the model Drake Passage, even by several thousand kilometers. The integral of the wind stress over the circumpolar streamlines is found to be a useful predictor of the magnitude of the volume transport through the model Drake Passage, although it is necessary to correct for basin-wide zonal pressure gradients in order to obtain good quantitative agreement