The Schistosoma mansoni Cytochrome P450 (CYP3050A1) Is Essential for Worm Survival and Egg Development.

Schistosomiasis affects millions of people in developing countries and is responsible for more than 200,000 deaths annually. Because of toxicity and limited spectrum of activity of alternatives, there is effectively only one drug, praziquantel, available for its treatment. Recent data suggest that drug resistance could soon be a problem. There is therefore the need to identify new drug targets and develop drugs for the treatment of schistosomiasis. Analysis of the Schistosoma mansoni genome sequence for proteins involved in detoxification processes found that it encodes a single cytochrome P450 (CYP450) gene. Here we report that the 1452 bp open reading frame has a characteristic heme-binding region in its catalytic domain with a conserved heme ligating cysteine, a hydrophobic leader sequence present as the membrane interacting region, and overall structural conservation. The highest sequence identity to human CYP450s is 22%. Double stranded RNA (dsRNA) silencing of S. mansoni (Sm)CYP450 in schistosomula results in worm death. Treating larval or adult worms with antifungal azole CYP450 inhibitors results in worm death at low micromolar concentrations. In addition, combinations of SmCYP450-specific dsRNA and miconazole show additive schistosomicidal effects supporting the hypothesis that SmCYP450 is the target of miconazole. Treatment of developing S. mansoni eggs with miconazole results in a dose dependent arrest in embryonic development. Our results indicate that SmCYP450 is essential for worm survival and egg development and validates it as a novel drug target. Preliminary structure-activity relationship suggests that the 1-(2,4-dichlorophenyl)-2-(1H-imidazol-1-yl)ethan-1-ol moiety of miconazole is necessary for activity and that miconazole activity and selectivity could be improved by rational drug design

Investigation of variable aeration of monodisperse mixtures: implications for pyroclastic density currents

© 2018, The Author(s). The high mobility of dense pyroclastic density currents (PDCs) is commonly attributed to high gas pore pressures. However, the influence of spatial and temporal variations in pore pressure within PDCs has yet to be investigated. Theory suggests that variability in the fluidisation and aeration of a current will have a significant control on PDC flow and deposition. In this study, the effect of spatially heterogeneous gas pore pressures in experimental PDCs was investigated. Sustained, unsteady granular currents were released into a flume channel where the injection of gas through the channel base was controlled to create spatial variations in aeration. Maximum current front velocity results from high degrees of aeration proximal to the source, rather than lower sustained aeration along the whole flume channel. However, moderate aeration (i.e. ~ 0.5 minimum static fluidisation velocity (U mf_st )) sustained throughout the propagation length of a current results in greater runout distances than currents which are closer to fluidisation (i.e. 0.9 U mf_st ) near to source, then de-aerating distally. Additionally, although all aerated currents are sensitive to channel base slope angle, the runout distance of those currents where aeration is sustained throughout their lengths increases by up to 54% with an increase of slope from 2° to 4°. Deposit morphologies a primarily controlled by the spatial differences in aeration, where there is a large decrease in aeration the current forms a thick depositional wedge. Sustained gas-aerated granular currents are observed to be spontaneously unsteady, with internal sediment waves travelling at different velocities

Improved climate simulations through a stochastic parameterization of ocean eddies

In climate simulations, the impacts of the sub-grid scales on the resolved scales are conventionally represented using deterministic closure schemes, which assume that the impacts are uniquely determined by the resolved scales. Stochastic parameterization relaxes this assumption, by sampling the sub-grid variability in a computationally inexpensive manner. This study shows that the simulated climatological state of the ocean is improved in many respects by implementing a simple stochastic parameterization of ocean eddies into a coupled atmosphere–ocean general circulation model. Simulations from a high-resolution, eddy-permitting ocean model are used to calculate the eddy statistics needed to inject realistic stochastic noise into a low-resolution, non-eddy-permitting version of the same model. A suite of four stochastic experiments is then run to test the sensitivity of the simulated climate to the noise definition, by varying the noise amplitude and decorrelation time within reasonable limits. The addition of zero-mean noise to the ocean temperature tendency is found to have a non-zero effect on the mean climate. Specifically, in terms of the ocean temperature and salinity fields both at the surface and at depth, the noise reduces many of the biases in the low-resolution model and causes it to more closely resemble the high-resolution model. The variability of the strength of the global ocean thermohaline circulation is also improved. It is concluded that stochastic ocean perturbations can yield reductions in climate model error that are comparable to those obtained by refining the resolution, but without the increased computational cost. Therefore, stochastic parameterizations of ocean eddies have the potential to significantly improve climate simulations

Design & manufacture of a high-performance bicycle crank by additive manufacturing

A new practical workflow for the laser Powder Bed Fusion (PBF) process, incorporating topological design, mechanical simulation, manufacture, and validation by computed tomography is presented, uniquely applied to a consumer product (crank for a high-performance racing bicycle), an approach that is tangible and adoptable by industry. The lightweight crank design was realised using topology optimisation software, developing an optimal design iteratively from a simple primitive within a design space and with the addition of load boundary conditions (obtained from prior biomechanical crank force–angle models) and constraints. Parametric design modification was necessary to meet the Design for Additive Manufacturing (DfAM)considerations for PBF to reduce build time, material usage, and post-processing labour. Static testing proved performance close to current market leaders with the PBF manufactured crank found to be stiffer than the benchmark design (static load deflection of 7.0±0.5 mm c.f. 7.67mm for a Shimano crank at a competitive mass (155g vs. 175g). Dynamic mechanical performance proved inadequate, with failure at 2495±125cycles; the failure mechanism was consistent in both its form and location. This research is valuable and novel as it demonstrates a complete work flow from design, manufacture, post-treatment, and validation of a highly loaded PBF manufactured consumer component, offering practitioners a validated approach to the application of PBF for components with application outside of the accepted sectors (aerospace, biomedical, autosports, space, and power generation)

Rotation of the cluster of galaxies A2107

We present indications of rotation in the galaxy cluster A2107 by a method that searches for the maximum gradient in the velocity field in a flat disk-like model of a cluster. Galaxies from cumulative sub-samples containing more and more distant members from the cluster centre, are projected onto an axis passing through the centre and we apply a linear regression model on the projected distances $x$ and the line-of-sight velocities $V$. The axis with the maximum linear correlation coefficient $r_{max} = \max [ r(V,x) ]$ defines the direction of the maximum velocity gradient, and consequently it presents the major axis of the apparently elliptical cluster. Because the effects of rotation are subtle, we put strong emphasis on the estimation of the uncertainties of the results by implementing different bootstrap techniques. We have found the rotational effects are more strongly expressed from distances $0.26 \div 0.54$ Mpc from the cluster centre. The total virial mass of the cluster is (3.2\pm0.6)\times10^{14} {\cal M}_{\sun}, while the virial mass, corrected for the rotation, is (2.8\pm0.5) \times 10^{14}{\cal M}_{\sun}.Comment: 7 pages, 9 figures gzipped tar file. to be published in MNRA

Experimental protocol for biodiesel production with isolation of alkenones as coproducts from commercial Isochrysis algal biomass

© The Author(s), 2016. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Journal of Visualized Experiments 112 (2016): e54189, doi:10.3791/54189.The need to replace petroleum fuels with alternatives from renewable and more environmentally sustainable sources is of growing importance. Biomass-derived biofuels have gained considerable attention in this regard, however first generation biofuels from edible crops like corn ethanol or soybean biodiesel have generally fallen out of favor. There is thus great interest in the development of methods for the production of liquid fuels from domestic and superior non-edible sources. Here we describe a detailed procedure for the production of a purified biodiesel from the marine microalgae Isochrysis. Additionally, a unique suite of lipids known as polyunsaturated long-chain alkenones are isolated in parallel as potentially valuable coproducts to offset the cost of biodiesel production. Multi-kilogram quantities of Isochrysis are purchased from two commercial sources, one as a wet paste (80% water) that is first dried prior to processing, and the other a dry milled powder (95% dry). Lipids are extracted with hexanes in a Soxhlet apparatus to produce an algal oil ("hexane algal oil") containing both traditional fats (i.e., triglycerides, 46-60% w/w) and alkenones (16-25% w/w). Saponification of the triglycerides in the algal oil allows for separation of the resulting free fatty acids (FFAs) from alkenone-containing neutral lipids. FFAs are then converted to biodiesel (i.e., fatty acid methyl esters, FAMEs) by acid-catalyzed esterification while alkenones are isolated and purified from the neutral lipids by crystallization. We demonstrate that biodiesel from both commercial Isochrysis biomasses have similar but not identical FAME profiles, characterized by elevated polyunsaturated fatty acid contents (approximately 40% w/w). Yields of biodiesel were consistently higher when starting from the Isochrysis wet paste (12% w/w vs. 7% w/w), which can be traced to lower amounts of hexane algal oil obtained from the powdered Isochrysis product.This work was supported by the National Science Foundation (CHE-1151492), the Northwest Advanced Renewables Alliance (fellowship to J. Wilson-Peltier), and through a private donation from friends of WHOI

Decolorization improves the fuel properties of algal biodiesel from Isochrysis sp.

© The Author(s), 2016. This is the author's version of the work and is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Fuel 179 (2016): 229-234, doi:10.1016/j.fuel.2016.03.061.Results from the comprehensive fuel testing according to American Society for Testing and Materials International (ASTM) standards of an alkenone-free and decolorized biodiesel produced from the industrially grown marine microalgae Isochrysis sp. are presented. Fatty acid methyl ester (FAME) profiles of the non-decolorized and subsequently decolorized biodiesel fuels were nearly identical, yet the fuel properties were remarkably different. Significant positive impacts on the cetane number, kinematic viscosity, and lubricity were observed, indicating a potential deleterious effect of pigments like chlorophylls and pheophytins on these fuel properties. The decolorization process using montmorillonite K10 gave on average 90% mass recovery, and allowed for an otherwise unobtainable cloud point determination. Oxidative stability of the decolorized Isochrysis biodiesel remained well below the minimum prescribed in biodiesel standards due to elevated content of highly polyunsaturated fatty acids, however other values were in the range of those prescribed in the ASTM standards. Overall, decolorization improved the fuel properties of biodiesel from Isochrysis and may provide a path toward improved biodiesel fuels from other algal species.This work was supported by the National Science Foundation (CHE-1151492) and through a private donation from friends of WHOI.2017-03-2

Accessing monomers, surfactants, and the queen bee substance by acrylate cross-metathesis of long-chain alkenones

Author Posting. © The Author(s), 2017. This is the author's version of the work. It is posted here under a nonexclusive, irrevocable, paid-up, worldwide license granted to WHOI. It is made available for personal use, not for redistribution. The definitive version was published in Journal of the American Oil Chemists' Society 94 (2017): 831-840, doi: 10.1007/s11746-017-2997-8.Polyunsaturated long-chain alkenones are a unique class of lipids biosynthesized in significant quantities (up to 20% of cell carbon) by several algae including the industrially grown marine microalgae Isochrysis. Alkenone structures are characterized by a long linear carbon-chain (35-40 carbons) with one to four trans-double bonds and terminating in a methyl or ethyl ketone. Alkenones were extracted and isolated from commercially obtained Isochrysis biomass and then subjected to cross-metathesis (CM) with methyl acrylate or acrylic acid using the Hoveyda-Grubbs metathesis initiator. Within 1 h at room temperature alkenones were consumed, however complete fragmentation (i.e. conversion to the smallest subunits by double bond cleavage) required up to 16 h. Analysis of the reaction mixture by gas chromatography and comprehensive two-dimensional gas chromatography revealed a predictable product mixture consisting primarily of long-chain (mostly C17) acids (or methyl esters from CM with methyl acrylate) and diacids (or diesters), along with smaller amounts (~5%) of the honey bee “queen substance” (E)-9-oxo-decenoic acid. Together, these compounds comprise a diverse mixture of valuable chemicals that includes surfactants, monomers, and an agriculturally relevant bee pheromone.This work was supported by the National Science Foundation (CHE- 1151492) and through a private donation from friends of WHOI