Substrate specificity and the effect of calcium on Trypanosomabrucei metacaspase 2

Metacaspases are cysteine peptidases found only in yeast, plants and lower eukaryotes, including the protozoa. To investigate the extended substrate specificity and effects of Ca<sup>2+</sup> on the activation of these enzymes, detailed kinetic, biochemical and structural analyses were carried out on metacaspase 2 from Trypanosoma brucei (TbMCA2). These results reveal that TbMCA2 has an unambiguous preference for basic amino acids at the P<sub>1</sub> position of peptide substrates and that this is most probably a result of hydrogen bonding from the P<sub>1</sub> residue to Asp95 and Asp211 in TbMCA2. In addition, TbMCA2 also has a preference for charged residues at the P<sub>2</sub> and P<sub>3</sub>positions and for small residues at the prime side of a peptide substrate. Studies into the effects of Ca<sup>2+</sup> on the enzyme revealed the presence of two Ca<sup>2+</sup> binding sites and a reversible structural modification of the enzyme upon Ca<sup>2+</sup> binding. In addition, the concentration of Ca<sup>2+</sup> used for activation of TbMCA2 was found to produce a differential effect on the activity of TbMCA2, but only when a series of peptides that differed in P<sub>2</sub> were examined, suggesting that Ca<sup>2+</sup>activation of TbMCA2 has a structural effect on the enzyme in the vicinity of the S2 binding pocket. Collectively, these data give new insights into the substrate specificity and Ca<sup>2+</sup> activation of TbMCA2. This provides important functional details and leads to a better understanding of metacaspases, which are known to play an important role in trypanosomes and make attractive drug targets due to their absence in humans

SERMeQ Model Produces a Realistic Upper Bound on Calving Retreat for 155 Greenland Outlet Glaciers

The rate of land ice loss due to iceberg calving is a key source of variability among model projections of the 21st century sea level rise. It is especially challenging to account for mass loss due to iceberg calving in Greenland, where ice drains to the ocean through hundreds of outlet glaciers, many smaller than typical model grid scale. Here, we apply a numerically efficient network flowline model (SERMeQ) forced by surface mass balance to simulate an upper bound on decadal calving retreat of 155 grounded outlet glaciers of the Greenland Ice Sheet—resolving five times as many outlets as was previously possible. We show that the upper bound holds for 91% of glaciers examined and that simulated changes in terminus position correlate with observed changes. SERMeQ can provide a physically consistent constraint on forward projections of the dynamic mass loss from the Greenland Ice Sheet associated with different climate projections.Key PointsWe test an upper‐bound model of calving retreat of 155 ocean‐terminating outlet glaciers that drain the Greenland Ice SheetOur physics‐based method produces terminus positions that correlate with observed positions for 103 glaciers without model tuningOur model bounds retreat rates on 91% of glaciers tested, providing a constraint for future sea level projectionsPeer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/163401/3/grl61420-sup-0003-2020GL090213-Text_SI-S01.pdfhttp://deepblue.lib.umich.edu/bitstream/2027.42/163401/2/grl61420_am.pdfhttp://deepblue.lib.umich.edu/bitstream/2027.42/163401/1/grl61420.pd

The effect of consumption volume on profile and liking of oral nutritional supplements of varied sweetness: sequential profiling and boredom tests

Oral nutrition supplements (ONS) are routinely prescribed to those with, or at risk of, malnutrition. Previous research identified poor compliance due to taste and sweetness. This paper investigates taste and hedonic liking of ONS, of varying sweetness and metallic levels, over consumption volume; an important consideration as patients are prescribed large volumes of ONS daily. A sequential descriptive profile was developed to determine the perception of sensory attributes over repeat consumption of ONS. Changes in liking of ONS following repeat consumption were characterised by a boredom test. Certain flavour (metallic taste, soya milk flavour) and mouthfeel (mouthdrying, mouthcoating) attributes built up over increased consumption volume (p 0.002). Hedonic liking data from two cohorts, healthy older volunteers (n = 32, median age 73) and patients (n = 28, median age 85), suggested such build-up was disliked. Efforts made to improve the palatability of ONS must take account of the build up of taste and mouthfeel characteristics over increased consumption volume

Classifying the embedded young stellar population in Perseus and Taurus & the LOMASS database

Context. The classification of young stellar objects (YSOs) is typically done using the infrared spectral slope or bolometric temperature, but either can result in contamination of samples. More accurate methods to determine the evolutionary stage of YSOs will improve the reliability of statistics for the embedded YSO population and provide more robust stage lifetimes. Aims. We aim to separate the truly embedded YSOs from more evolved sources. Methods. Maps of HCO+ J=4-3 and C18O J=3-2 were observed with HARP on the James Clerk Maxwell Telescope (JCMT) for a sample of 56 candidate YSOs in Perseus and Taurus in order to characterize emission from high (column) density gas. These are supplemented with archival dust continuum maps observed with SCUBA on the JCMT and Herschel PACS to compare the morphology of the gas and dust in the protostellar envelopes. The spatial concentration of HCO+ J=4-3 and 850 micron dust emission are used to classify the embedded nature of YSOs. Results. Approximately 30% of Class 0+I sources in Perseus and Taurus are not Stage I, but are likely to be more evolved Stage II pre-main sequence (PMS) stars with disks. An additional 16% are confused sources with an uncertain evolutionary stage. Conclusions. Separating classifications by cloud reveals that a high percentage of the Class 0+I sources in the Perseus star forming region are truly embedded Stage I sources (71%), while the Taurus cloud hosts a majority of evolved PMS stars with disks (68%). The concentration factor method is useful to correct misidentified embedded YSOs, yielding higher accuracy for YSO population statistics and Stage timescales. Current estimates (0.54 Myr) may overpredict the Stage I lifetime on the order of 30%, resulting in timescales of 0.38 Myr for the embedded phase.Comment: 33 pages, 21 figures, 6 tables, Accepted to be published in A&

Sensory and instrumental analysis of medium and long shelf-life Charentais cantaloupe melons (Cucumis melo L.) harvested at different maturities

The flavour profiles of two genotypes of Charentais cantaloupe melons (medium shelf-life and long shelf-life), harvested at two distinct maturities (immature and mature fruit), were investigated. Dynamic headspace extraction (DHE), solid-phase extraction (SPE), gas chromatography–mass spectrometry (GC-MS) and gas chromatography–olfactometry/mass spectrometry (GC-O/MS) were used to determine volatile and semi-volatile compounds. Qualitative descriptive analysis (QDA) was used to assess the organoleptic impact of the different melons and the sensory data were correlated with the chemical analysis. There were significant, consistent and substantial differences between the mature and immature fruit for the medium shelf-life genotype, the less mature giving a green, cucumber character and lacking the sweet, fruity character of the mature fruit. However, maturity at harvest had a much smaller impact on the long shelf-life melons and fewer differences were detected. These long shelf-life melons tasted sweet, but lacked fruity flavours, instead exhibiting a musty, earthy character

The RMS Survey: Ammonia and water maser analysis of massive star forming regions

The Red MSX Source (RMS) survey has identified a sample of ~1200 massive young stellar objects (MYSOs), compact and ultra compact HII regions from a sample of ~2000 MSX and 2MASS colour selected sources. We have used the 100 m Green Bank telescope to search for 22-24 GHz water maser and ammonia (1,1), (2,2) and (3,3) emission towards ~600 RMS sources located within the northern Galactic plane. We have identified 308 H2O masers which corresponds to an overall detection rate of ~50%. Abridged: We detect ammonia emission towards 479 of these massive young stars, which corresponds to ~80%. Ammonia is an excellent probe of high density gas allowing us to measure key parameters such as gas temperatures, opacities, and column densities, as well as providing an insight into the gas kinematics. The average kinetic temperature, FWHM line width and total NH3 column density for the sample are approximately 22 K, 2 km/s and 2x10^{15} cm^{-2}, respectively. We find that the NH3 (1,1) line width and kinetic temperature are correlated with luminosity and finding no underlying dependence of these parameters on the evolutionary phase of the embedded sources, we conclude that the observed trends in the derived parameters are more likely to be due to the energy output of the central source and/or the line width-clump mass relationship. The velocities of the peak H2O masers and the NH3 emission are in excellent agreement with each other, which would strongly suggest an association between the dense gas and the maser emission. Moreover, we find the bolometric luminosity of the embedded source and the isotropic luminosity of the H2O maser are also correlated. We conclude from the correlations of the cloud and water maser velocities and the bolometric and maser luminosity that there is a strong dynamical relationship between the embedded young massive star and the H2O maser.Comment: 17 pages and 17 figures and 8 tables. Tables\,2 and 5 and full versions of Figs. 3 and 7 are only available in electronic form at the CDS via anonymous ftp to cdsarc.u-strasbg.fr (130.79.125.5) or via http://cdsweb.u-strasbg.fr/cgi-bin/qcat?J/A+A

High-throughput screening with the Eimeria tenella CDC2-related kinase2/cyclin complex EtCRK2/EtCYC3a

The poultry disease coccidiosis, caused by infection with Eimeria spp. apicomplexan parasites, is responsible for enormous economic losses to the global poultry industry. The rapid increase of resistance to therapeutic agents, as well as the expense of vaccination with live attenuated vaccines, requires the development of new effective treatments for coccidiosis. Because of their key regulatory function in the eukaryotic cell cycle, cyclin-dependent kinases (CDKs) are prominent drug targets. The Eimeria tenella CDC2-related kinase 2 (EtCRK2) is a validated drug target that can be activated in vitro by the CDK activator XlRINGO (Xenopus laevis rapid inducer of G2/M progression in oocytes). Bioinformatics analyses revealed four putative E. tenella cyclins (EtCYCs) that are closely related to cyclins found in the human apicomplexan parasite Plasmodium falciparum. EtCYC3a was cloned, expressed in Escherichia coli and purified in a complex with EtCRK2. Using the non-radioactive time-resolved fluorescence energy transfer (TR-FRET) assay, we demonstrated the ability of EtCYC3a to activate EtCRK2 as shown previously for XlRINGO. The EtCRK2/EtCYC3a complex was used for a combined in vitro and in silico high-throughput screening approach, which resulted in three lead structures, a naphthoquinone, an 8-hydroxyquinoline and a 2-pyrimidinyl-aminopiperidine-propane-2-ol. This constitutes a promising starting point for the subsequent lead optimization phase and the development of novel anticoccidial drugs

Deformation of a nearly hemispherical conducting drop due to an electric field: theory and experiment

We consider, both theoretically and experimentally, the deformation due to an electric field of a pinned nearly-hemispherical static sessile drop of an ionic fluid with a high conductivity resting on the lower substrate of a parallel plate capacitor. Using both numerical and asymptotic approaches we find solutions to the coupled electrostatic and augmented Young–Laplace equations which agree very well with the experimental results. Our asymptotic solution for the drop interface extends previous work in two ways, namely to drops that have zero-field contact angles that are not exactly π/2 and to higher order in the applied electric field, and provides useful predictive equations for the changes in the height, contact angle and pressure as functions of the zero-field contact angle, drop radius, surface tension and applied electric field. The asymptotic solution requires some numerical computations, and so a surprisingly accurate approximate analytical asymptotic solution is also obtained

Transient flow-driven distortion of a nematic liquid crystal in channel flow with dissipative weak planar anchoring

Motivated by the one-drop-filling (ODF) method for the industrial manufacturing of liquid crystal displays, we analyze the pressure-driven flow of a nematic in a channel with dissipative weak planar anchoring at the boundaries of the channel. We obtain quasisteady asymptotic solutions for the director angle and the velocity in the limit of small Leslie angle, in which case the key parameters are the Ericksen number and the anchoring strength parameter. In the limit of large Ericksen number, the solution for the director angle has narrow reorientational boundary layers and a narrow reorientational internal layer separated by two outer regions in which the director is aligned at the positive Leslie angle in the lower half of the channel and the negative Leslie angle in the upper half of the channel. On the other hand, in the limit of small Ericksen number, the solution for the director angle is dominated by splay elastic effects with viscous effects appearing at first order. As the Ericksen number varies, there is a continuous transition between these asymptotic behaviors, and in fact the two asymptotic solutions capture the behavior rather well for all values of the Ericksen number. The steady-state value of the director angle at the boundaries and the timescale of the evolution toward this steady-state value in the asymptotic limits of large and small Ericksen number are determined. In particular, using estimated parameter values for the ODF method, it is found that the boundary director rotation timescale is substantially shorter than the timescale of the ODF method, suggesting that there is sufficient time for significant transient flow-driven distortion of the nematic molecules at the substrates from their required orientation to occur