Native Mass Spectrometry for the Study of PROTAC GNE-987-Containing Ternary Complexes

PRO teolysis TA rgeting C himeras (PROTACs) promote the degradation, rather than inhibition, of a drug target as a mechanism for therapeutic treatment. Bifunctional PROTAC molecules allow simultaneous binding of both the target protein and an E3-Ubiquitin ligase, bringing the two proteins into close spatial proximity to allow ubiquitinylation and degradation of the target protein via the cell's endogenous protein degradation pathway. We utilized native mass spectrometry (MS) to study the ternary complexes promoted by the previously reported PROTAC GNE-987 between Brd4 bromodomains 1 and 2, and Von Hippel Lindeau E3-Ubiquitin Ligase. Native MS at high resolution allowed us to measure ternary complex formation as a function of PROTAC concentration to provide a measure of complex affinity and stability, whilst simultaneously measuring other intermediate protein species. Native MS provides a high-throughput, low sample consumption, direct screening method to measure ternary complexes for PROTAC development.     

Multipass Scratch Behavior of Borided and Nitrided H13 Steel

AISI H13 steel was subjected to boriding and nitriding; its performance was evaluated by the progressive load scratch test, and then by the multipass scratch test (MPST), to evaluate the accumulated damage. Boriding and nitriding were carried out at a temperature of 800 and 580 °C, respectively, both treatments for 1 and 5 h of exposure time. In scratch test, a load range of 3-90 N was used for borides; a higher range (3-180 N) was chosen for nitrides. Critical loads (L_c) were estimated based on optical microscope observations of the scratch tracks. Failure mechanisms and residual depths were examined by scanning electronic microscope and optical profilometry. Finally, using fractions of L_c, MPST was applied on uncoated and coated samples, for 25, 50, 75 and 100 unidirectional scratch cycles. The coefficient of friction (COF) evolution was recorded and analyzed. A decrease in COF was observed as cycles go through; it was initially higher in uncoated sample; moreover, it was observed that normal load was determinant in its behavior, regardless of the treatment time. Nitrided samples exhibited less catastrophic failures over borided ones, also showed a better volume loss/load ratio performance, from 6 to 8 times better compared to borided samples.     

Potential Applications of Concentrated Solar Thermal Technologies in the Australian Minerals Processing and Extractive Metallurgical Industry

The Australian minerals processing and extractive metallurgy industries are responsible for about 20% of Australia’s total greenhouse gas (GHG) emissions. This article reviews the potential applications of concentrated solar thermal (CST) energy in the Australian minerals processing industry to reduce this impact. Integrating CST energy into these industries would reduce their reliance upon conventional fossil fuels and reduce GHG emissions. As CST technologies become more widely deployed and cheaper, and as fuel prices rise, CST energy will progressively become more competitive with conventional energy sources. Some of the applications identified in this article are expected to become commercially competitive provided the costs for pollution abatement and GHG mitigation are internalized. The areas of potential for CST integration identified in this study can be classed as either medium/low-temperature or high-temperature applications. The most promising medium/low-grade applications are electricity generation and low grade heating of liquids. Electricity generation with CST energy—also known as concentrated solar power—has the greatest potential to reduce GHG emissions out of all the potential applications identified because of the 24/7 dispatchability when integrated with thermal storage. High-temperature applications identified include the thermal decomposition of alumina and the calcination of limestone to lime in solar kilns, as well as the production of syngas from natural gas and carbonaceous materials for various metallurgical processes including nickel and direct reduced iron production. Hybridization and integration with thermal storage could enable CST to sustain these energy-intensive metallurgical processes continuously. High-temperature applications are the focus of this paper.     

Modelling of the disulphide-swapped isomer of human insulin-like growth factor-1: implications for receptor binding

Insulin-like growth factor-1 (IGF-1) is a serum protein whichunexpectedly folds to yield two stable tertiary structures withdifferent disulphide connectivities; native IGF-1 [18–61,6–48,47–52]and IGF-1 swap [18–61,6–47, 48–52]. Here we demonstratein detail the biological properties of recombinant human nativeIGF-1 and IGF-1 swap secreted from Saccharomyces cerevisiae.IGF-1 swap had a ~30 fold loss in affinity for the IGF-1 receptoroverexpressed on BHK cells compared with native IGF-1.The parallelincrease in dose required to induce negative cooperativity togetherwith the parallel loss in mitogenicity in NIH 3T3 cells impliesthat disruption of the IGF-1 receptor binding interaction ratherthan restriction of a post-binding conformational change isresponsible for the reduction in biological activity of IGF-1swap. Interestingly, the affinity of IGF-1 swap for the insulinreceptor was ~200 fold lower than that of native IGF-1 indicatingthat the binding surface complementary to the insulin receptor(or the ability to attain it) is disturbed to a greater extentthan that to the IGF-1 receptor. A 1.0 ns high-temperature moleculardynamics study of the local energy landscape of IGF-1 swap resultedin uncoiling of the first A-region -helix and a rearrangementin the relative orientation of the A- and B-regions. The modelof IGF-1 swap is structurally homologous to the NMR structureof insulin swap and CD spectra consistent with the model arepresented. However, in the model of IGF-1 swap the C-regionhas filled the space where the first A-region -helix has uncoiledand this may be hindering interaction of Val44 with the secondinsulin receptor binding pocket.     

Landfill Risk Assessment on Groundwater Based on Vulnerability and Pollution Index

Many existing risk assessment methodologies heavily rely on complex physical models and probabilistic concepts, which are difficult to use and not suitable for a first-cut analysis. Meanwhile, most groundwater pollution risk assessment indices generally are solely based on groundwater vulnerability and ignore the consideration of the hazard of pollution source. In this paper, we developed a simple groundwater pollution risk index approach, which can be easily utilized in many developing countries. The approach is developed according to vulnerability of the aquifers and pollution source index, which is based on landfill treatment technology. Applications of this risk index approach to 22 formal landfill sites in Beijing show that 1, 2, 4, 10, 5 landfill sites can be classified as of high, medium high, medium, medium low, low risk level respectively. These results demonstrate that vulnerability assessment alone is not sufficient to groundwater pollution risk assessment. This simple risk assessment method is most suitable for a comparative assessment of a large number of pollution sources at a regional scale due to its lower cost and easy operability. It also allows prioritizes landfills for reclamation interventions and recommendation of required actions such that an efficient management of landfills could be accomplished.     

Sulfamethazine water medication pharmacokinetics and contamination in a commercial pig production unit

Sulfamethazine is often used to treat disease in the swine industry. Sulfamethazine is available as water or feed medication and historically (over the past 40 years) has been associated with residue violations in both the United States and Europe. Despite sulfamethazine's approval for use as a water medication, little research on the pharmacokinetics of the water formulation is available. Therefore, a pilot study was performed to determine the plasma levels of an approved sulfamethazine water medication. Plasma levels in pigs treated with an oral bolus (250 mg/kg), which is equivalent to the total drug consumed within a 24-h period, achieved therapeutic concentrations (50 microg/ml). Noncompartmental-based pharmacokinetic model parameters for clearance, half-life, and volume of distribution were consistent with previously published values in swine. However, the above treatment resulted in exposure of pen mates to sulfamethazine at levels currently above tolerance (0.1 ppm). Using a physiologically based pharmacokinetic model, the treatment dose simulation was compared with observed plasma levels of treated pigs. Flexibility of the physiologically based pharmacokinetic model also allowed simulation of control-pig plasma levels to estimate contamination exposure. A simulated exposure to 0.15 mg/kg twice within approximately 8 h resulted in detectable levels of sulfamethazine in the control pigs. After initial exposure, a much lower dose of 0.059 mg/kg maintained the contamination levels above tolerance for at least 3 days. These results are of concern for producers and veterinarians, because in commercial farms, the entire barn is often treated,and environmental contamination could result in residues of an unknown duration.     

Annual redd counts across two decades are indicators of changing migratory bull trout population sizes in two proximate Montana creeks

Redd counts are the most widely used method for bull trout (Salvelinus confluentus Suckley) population monitoring. Redd counts are an assumed indicator of population size (N), particularly for female spawners, but previous studies rarely considered the measurement uncertainty of N. The present study of two proximate, migratory bull trout populations across two decades examined statistical correlations between annual redd counts and concurrent N (predominantly fish ages 3–7) that included measurement uncertainty. Results showed that redd counts explained about one-third of the concurrent N variation and, in one case, portended N 7 years later. But changing spawning distributions across years, as likewise shown here and as a probable effect of ongoing climate change, can confound the use of fixed index reaches for redd counts. Instead, abundance monitoring based on redd counts should be over a bull trout population's or metapopulation's entire spatial extent and have counts in all potential spawning areas. Such total-annual redd counts may be the most cost-effective and precise means of bull trout population monitoring.     

快速煅烧分解石灰石对其吸收CO2和SO2的影响

在加热过程中,应用一种新型的快速加热装置——铂膜反应器,及时准确地获得样品质量和颗粒温度,来研究石灰石的煅烧分解,并对石灰石加热速率高达750℃/s下的分解反应特性,及其对分解后产生的氧化钙吸收CO2和脱硫的化学反应特性进行了研究.实验表明,在初始加热阶段,高升温速率明显阻碍了煅烧分解的进行,这对分解后的CaO吸收CO2的效果影响不大.当加热速率升高时,石灰石分解后产生的CaO吸收CO2和脱硫过程中,钙利用率显著下降.     

In situ metal ion contamination and the effects on proton exchange membrane fuel cell performance

Automotive fuel cell technology has made considerable progress, and hydrogen fuel cell vehicles are regarded as a possible long-term solution to reduce carbon dioxide emissions, reduce fossil fuel dependency and increase energy efficiency. Even though great strides have been made, durability is still an issue. One key challenge is controlling MEA contamination. Metal ion contamination within the membrane and the effects on fuel cell performance were investigated. Given the possible benefits of using stainless steel or aluminum for balance-of-plant components or bipolar plates, cations of Al, Fe, Ni and Cr were studied. Membranes were immersed in metal sulfide solutions of varying concentration and then assembled into fuel cell MEAs tested in situ. The ranking of the four transition metals tested in terms of the greatest reduction in fuel cell performance was: Al³⁺ ? Fe²⁺ > Ni²⁺, Cr³⁺. For iron-contaminated membranes, no change in cell performance was detected until the membrane conductivity loss was greater than approximately 15%.