Processing polymer photocatalysts for photocatalytic hydrogen evolution.

Conjugated materials have emerged as competitive photocatalysts for the production of sustainable hydrogen from water over the last decade. Interest in these polymer photocatalysts stems from the relative ease to tune their electronic properties through molecular engineering, and their potentially low cost. However, most polymer photocatalysts have only been utilised in rudimentary suspension-based photocatalytic reactors, which are not scalable as these systems can suffer from significant optical losses and often require constant agitation to maintain the suspension. Here, we will explore research performed to utilise polymeric photocatalysts in more sophisticated systems, such as films or as nanoparticulate suspensions, which can enhance photocatalytic performance or act as a demonstration of how the polymer can be scaled for real-world applications. We will also discuss how the systems were prepared and consider both the benefits and drawbacks of each system before concluding with an outlook on the field of processable polymer photocatalysts.

Meteorite evidence for partial differentiation and protracted accretion of planetesimals.

Modern meteorite classification schemes assume that no single planetary body could be source of both unmelted (chondritic) and melted (achondritic) meteorites. This dichotomy is a natural outcome of formation models assuming that planetesimal accretion occurred nearly instantaneously. However, it has recently been proposed that the accretion of many planetesimals lasted over ≳1 million years (Ma). This could have resulted in partially differentiated internal structures, with individual bodies containing iron cores, achondritic silicate mantles, and chondritic crusts. This proposal can be tested by searching for a meteorite group containing evidence for these three layers. We combine synchrotron paleomagnetic analyses with thermal, impact, and collisional evolution models to show that the parent body of the enigmatic IIE iron meteorites was such a partially differentiated planetesimal. This implies that some chondrites and achondrites simultaneously coexisted on the same planetesimal, indicating that accretion was protracted and that apparently undifferentiated asteroids may contain melted interiors.

Comparing Single-Objective Optimization Protocols for Calibrating the Birds Nest Aquifer Model-A Problem Having Multiple Local Optima.

To best represent reality, simulation models of environmental and health-related systems might be very nonlinear. Model calibration ideally identifies globally optimal sets of parameters to use for subsequent prediction. For a nonlinear system having multiple local optima, calibration can be tedious. For such a system, we contrast calibration results from PEST, a commonly used automated parameter estimation program versus several meta-heuristic global optimizers available as external packages for the Python computer language-the Gray Wolf Optimization (GWO) algorithm; the DYCORS optimizer framework with a Radial Basis Function surrogate simulator (DRB); and particle swarm optimization (PSO). We ran each optimizer 15 times, with nearly 10,000 MODFLOW simulations per run for the global optimizers, to calibrate a steady-state, groundwater flow simulation model of the complex Birds Nest aquifer, a three-layer system having 8 horizontal hydraulic conductivity zones and 25 head observation locations. In calibrating the eight hydraulic conductivity values, GWO averaged the best root mean squared error (RMSE) between observed and simulated heads-20 percent better (lower) than the next lowest optimizer, DRB. The best PEST run matched the best GWO RMSE, but both the average PEST RMSE and the range of PEST RMSE results were an order of magnitude larger than any of the global optimizers.

How desirable are your IC50s? A way to enhance screening-based decision making.

Dose-response curves, resulting in estimates of endpoints such as the IC(50), are fundamental to drug discovery. However, some estimates are more reliable than others. It is important to know just how reliable an estimate is if we want to base decisions on it or use it in further modeling. In this study, the authors propose a new measure of endpoint reliability, based on the concept of desirability first introduced by Harrington. The solution is not dependent on the application used to analyze the experimental data, provided a number of parameters to characterize the dose-response curve are available. The authors show how this score can be used as an objective and consistent measure to rank screening results, combine information from groups of experiments, and determine optimal levels of characterization of a compound's biological activity.

An automated, high-throughput, 384 well Cytochrome P450 cocktail IC50 assay using a rapid resolution LC-MS/MS end-point.

The current study focused on the development of an automated IC50 cocktail assay in a miniaturized 384 well assay format. This was developed in combination with a significantly shorter high pressure liquid chromatography (HPLC) separation and liquid chromatography-mass spectrometry (LC-MS/MS) run-time; than those currently reported in the literature. The 384-well assay used human liver microsomes in conjunction with a cocktail of probe substrates metabolized by the five major CYPs (tacrine for CYP1A2, diclofenac for CYP2C9, (S)-mephenytoin for CYP2C19, dextromethorphan for CYP2D6 and midazolam for CYP3A4). To validate the usefulness of the automated and analytical methodologies, IC50 determinations were performed for a series of test compounds known to exhibit inhibition across these five major P450s. Eight compounds (sertraline, disulfuram, ticlopidine fluconazole, fluvoxamine, ketoconazole, miconazole, paroxetine, flunitrazepam) were studied as part of a cocktail assay, and against each CYPs individually. The data showed that the IC50s generated with cocktail incubations did not differ to a great extent from those obtained in the single probe experiments and hence unlikely to significantly influence the predicted clinical DDI risk. In addition the present method offered a significant advantage over some of the existing cocktail analytical methodology in that separation can be achieved with run times as short as 1 min without compromising data integrity. Although numerous studies have been reported to measure CYP inhibition in a cocktail format the need to support growing discovery libraries not only relies on higher throughput assays but quicker analytical run times. The current study reports a miniaturized high-throughput cocktail IC50 assay, in conjunction with a robust, rapid resolution LC-MS/MS end-point offered increased sample throughput without compromising analytical sensitivity or analyte resolution.

A three-stage experimental strategy to evaluate and validate an interplate IC50 format.

The serial dilution of compounds to establish potency against target enzymes or receptors can at times be a rate-limiting step in project progression. We have investigated the possibility of running 50% inhibitory concentration experiments in an interplate format, with dose ranges constructed across plates. The advantages associated with this format include a faster reformatting time for the compounds while also increasing the number of doses that can be potentially generated. These two factors, in particular, would lend themselves to a higher-throughput and more timely testing of compounds, while also maximizing chances to capture fully developed dose-response curves. The key objective from this work was to establish a strategy to assess the feasibility of an interplate format to ensure that the quality of data generated would be equivalent to historical formats used. A three-stage approach was adopted to assess and validate running an assay in an interplate format, compared to an intraplate format. Although the three-stage strategy was tested with two different assay formats, it would be necessary to investigate the feasibility for other assay types. The recommendation is that the three-stage experimental strategy defined here is used to assess feasibility of other assay formats used.

Tetravalent neutralizing antibody response against four dengue serotypes by a single chimeric dengue envelope antigen.

We employed DNA shuffling and screening technologies to develop a single recombinant dengue envelope (E) antigen capable of inducing neutralizing antibodies against all four antigenically distinct dengue serotypes. By DNA shuffling of codon-optimized dengue 1-4 E genes, we created a panel of novel chimeric clones expressing C-terminal truncated E antigens that combined epitopes from all four dengue serotypes. DNA vaccines encoding these novel chimeras induced multivalent T cell and neutralizing antibody responses against all four dengue serotypes in mice. By contrast, a mixture of four unshuffled, parental DNA vaccines failed to produce tetravalent neutralizing antibodies in mice. The neutralizing antibody titers for some of these antigens could be further improved by extending the sequences to express full-length pre-membrane and envelope proteins. The chimeric antigens also protected mice against a lethal dengue-2 virus challenge. These data demonstrate that DNA shuffling and associated screening can lead to the selection of multi-epitope antigens against closely related dengue virus serotypes and suggest a broad utility for these technologies in optimizing vaccine antigens.

Evaluating real-life high-throughput screening data.

High-throughput screening (HTS) is the result of a concerted effort of chemistry, biology, information technology, and engineering. Many factors beyond the biology of the assay influence the quality and outcome of the screening process, yet data analysis and quality control are often focused on the analysis of a limited set of control wells and the calculated values derived from these wells. Taking into account the large number of variables and the amount of data generated, multiple views of the screening data are necessary to guarantee quality and validity of HTS results. This article does not aim to give an exhaustive outlook on HTS data analysis but tries to illustrate the shortfalls of a reductionist approach focused on control wells and give examples for further analysis.

An inducible nitric oxide synthase-luciferase reporter system for in vivo testing of anti-inflammatory compounds in transgenic mice.

The inducible NO synthase gene (iNOS) plays a role in a number of chronic and acute conditions, including septic shock and contact hypersensitivity autoimmune diseases, such as rheumatoid arthritis, gastrointestinal disorders, and myocardial ischemia. The iNOS gene is primarily under transcriptional control and is induced in a variety of conditions. The ability to monitor and quantify iNOS expression in vivo may facilitate a better understanding of the role of iNOS in different diseases. In this study, we describe a transgenic mouse (iNos-luc) in which the luciferase reporter is under control of the murine iNOS promoter. In an acute sepsis model produced by injection of IFN-gamma and LPS, we observed an induction of iNOS-driven luciferase activity in the mouse liver. This transgene induction is dose and time dependent and correlated with an increase of liver iNOS protein and iNOS mRNA levels. With this model, we tested 11 compounds previously shown to inhibit iNOS induction in vitro or in vivo. Administration of dexamethasone, epigallocatechin gallate, alpha-phenyl-N-tert-butyl nitrone, and ebselen significantly suppressed iNOS transgene induction by IFN-gamma and LPS. We further evaluated the use of the iNos-luc transgenic mice in a zymosan-induced arthritis model. Intra-articular injection of zymosan induced iNos-luc expression in the knee joint. The establishment of the iNos-luc transgenic model provides a valuable tool for studying processes in which the iNOS gene is induced and for screening anti-inflammatory compounds in vivo.