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Diets high in saturated fat (HFD) disrupt dopamine neurotransmission, whereas fasting alters tonic and phasic dopamine release to drive motivation and food consumption. However, functional compartments in the nucleus accumbens (NAc) influencing these effects are not well characterized, and sex comparisons have not been made. This study sought to determine whether consumption of a HFD, sex, or being fed versus fasted altered baseline dopamine release and reuptake throughout NAc subregions. Male and female C57BL/6 mice were fed a control diet or nutrient matched HFD for six weeks. Ex-vivo fast-scan cyclic voltammetry revealed females had significantly slower dopamine reuptake in the NAc core than males when fed ad lib control diet. Fasting enhanced dopamine release and reuptake in the NAc core but not the medioventral shell. Further, being fasted versus fed significantly increased dopamine release throughout the NAc core in control males but specifically promoted release and reuptake in only the ventrolateral core of HF-fed males, effects which were lacking in females. Finally, fasting promoted dopamine release and reuptake in the rostral NAc core of controls and more caudally in HFD groups. These data support that dopamine neurotransmission is heterogeneous in NAc subregions and suggest the ventrolateral core is responsive to energy state. Furthermore, a rostrocaudal gradient in the NAc core might control valence responses to fasting that could promote overeating after chronic HFD consumption.

Mass spectrometry (MS) is a ubiquitous method in analytical science, with applications ranging from physics to molecular biology. The fundamental nature of mass makes weighing molecules and/or their fragments a reliable way to characterize identity, structure, and function. Conventional MS methods work by measuring the mass to charge ratio (m/z) of an ensemble of ionized analyte molecules. The mass is then determined by multiplying the m/z by the charge (z) of the ion, a quantity that is typically determined either by ionization method or by measuring the spacing between m/z peaks. For smaller molecules, deconvolving the charge from the m/z spectrum is straightforward, but becomes increasingly difficult for large molecules (>1 MDa) or samples that contain heterogeneous mixtures because m/z peaks overlap and are unresolved. Single ion MS techniques circumvent this problem by measuring the mass of individual ions via independent measurements of both the m/z and the charge, but these measurement methods are typically too time-consuming to be practical for acquiring the thousands of ions necessary to produce meaningful mass spectrum. This dissertation focuses on the development and application of multiplexed charge detection mass spectrometry (CDMS), a single ion MS technique that enables much faster individual ion mass measurements. A novel data analysis method that uses the amplitudes of the first and second harmonic frequencies in the Fourier transform of ion signal to dynamically determine ion energies is demonstrated. By monitoring the ion energies of individually trapped ions in CDMS, information about ion cross section and neutral solvent evaporation is obtained. For the first time, multiplexed CDMS measurements are demonstrated and the time necessary to acquire a statistically significant sample of individual ion data is decreased by an order of magnitude compared to other CDMS techniques. Simulations of optimized CDMS data show that multiplexed measurements combined with a novel method for decoupling ion m/z and frequency can further decrease the ion acquisition time to less than one minute. These CDMS techniques are applied to the study of the mechanisms of electrospray ionization for nanometer-sized droplets and macromolecules originating from different aqueous solutions and new insights on these previously difficult to measure systems are gained.

N-terminal glutamate (E) cyclization to form pyroglutamate (pE) generates charge heterogeneities for mAbs and proteins. Thus far, pE formation rate in lyophilized formulation as compared to in liquid formulation has not been reported. Impact of pE on antibody biological activity has only been predicted or assessed using stressed samples that may contain other confounding degradations besides pE. Additionally, application of hydrophobic interaction chromatography (HIC) to separate pE has not been reported. In our study, N-terminal E cyclization was identified as the major degradation pathway in lyophilized formulation at elevated temperature for both monoclonal antibody (mAb-A) and IgG-like bispecific antibody (bsAb-A). pE was enriched in salt-gradient ion exchange chromatography (IEC) as pre-peak and in HIC as post-peak for both mAb-A and bsAb-A. Structure-function studies with pE-enriched IEC and HIC fractions confirmed that pE did not affect binding activities for mAb-A and bsAb-A. In vitro incubation of bsAb-A in serum and PBS revealed that the serum matrix may play a role in pE conversion in human serum, in contrast to the chemical reaction mechanism reported. These techniques can help in characterization of N-terminal E-to-pE cyclization and quality attribute severity assessment during therapeutic protein product development.

High-fat diets are linked with obesity and changes in dopamine neurotransmission. Mounting evidence shows that saturated fat impacts dopamine neurons and their terminal fields, but little is known about the effect a diet high in unsaturated fat has on the dopamine system. This study sought to determine whether fat type, saturated vs. unsaturated, differentially affected body weight, blood glucose regulation, locomotor behavior, and control of dopamine release and uptake at dopamine neuron terminals in the nucleus accumbens (NAc). C57BL/6 mice were fed a control diet or a nutrient-matched diet high in saturated fat (SF), unsaturated flaxseed oil (Flax) or a blend of the two fats. After 6-weeks, mice from each high-fat diet group gained significantly more weight than Controls, but the group fed Flax gained less weight than the SF group and had fasting blood glucose levels similar to Controls. Ex-vivo fast scan cyclic voltammetry revealed the SF group also had significantly slower synaptic dopamine clearance and a reduced capacity for phasic dopamine release in the nucleus accumbens (NAc), but the Flax and Blend groups resembled Controls. These data show that different types of dietary fat have substantially different effects on metabolic phenotype and influence how dopamine terminals in the NAc regulate dopamine neurotransmission. Our data also suggests that a diet high in unsaturated fat may preserve normal metabolic and behavioral parameters as well as dopamine signaling in the NAc.

Medical shortages during the COVID-19 pandemic saw numerous efforts to 3D print personal protective equipment and treatment supplies. There is, however, little research on the potential biocompatibility of 3D-printed parts using typical polymeric resins as pertaining to volatile organic compounds (VOCs), which have specific relevance for respiratory circuit equipment. Here, we measured VOCs emitted from freshly printed stereolithography (SLA) replacement medical parts using proton transfer reaction mass spectrometry and infrared differential absorption spectroscopy, and particulates using a scanning mobility particle sizer. We observed emission factors for individual VOCs ranging from ∼0.001 to ∼10 ng cm–3 min–1. Emissions were heavily dependent on postprint curing and mildly dependent on the type of SLA resin. Curing reduced the emission of all observed chemicals, and no compounds exceeded the recommended dose of 360 μg/d. VOC emissions steadily decreased for all parts over time, with an average e-folding time scale (time to decrease to 1/e of the starting value) of 2.6 ± 0.9 h.

Bivalent proteolysis-targeting chimeras (PROTACs) drive protein degradation by simultaneously binding a target protein and an E3 ligase and forming a productive ternary complex. We hypothesized that increasing binding valency within a PROTAC could enhance degradation. Here, we designed trivalent PROTACs consisting of a bivalent bromo and extra terminal (BET) inhibitor and an E3 ligand tethered via a branched linker. We identified von Hippel-Lindau (VHL)-based SIM1 as a low picomolar BET degrader with preference for bromodomain containing 2 (BRD2). Compared to bivalent PROTACs, SIM1 showed more sustained and higher degradation efficacy, which led to more potent anticancer activity. Mechanistically, SIM1 simultaneously engages with high avidity both BET bromodomains in a cis intramolecular fashion and forms a 1:1:1 ternary complex with VHL, exhibiting positive cooperativity and high cellular stability with prolonged residence time. Collectively, our data along with favorable in vivo pharmacokinetics demonstrate that augmenting the binding valency of proximity-induced modalities can be an enabling strategy for advancing functional outcomes.

Mass filter operation in higher stability zones is known to provide better resolution. Unfortunately, for sine driven instruments, higher stability zone operation reduces the accessible mass range and increases the degenerative effects of fringe fields. Conversely, digitally driven mass filters do not suffer from loss of mass range, and the fringe field effects do not increase significantly by switching stability zones because the AC voltage is always constant and the DC voltage is always zero. This work catalogues 12 stability zones that are accessible with the new digital waveform generation technology. These zones have theoretical baseline resolving powers that range from 22 to 1 300 000 with pseudopotential well depths that range from 3.5 to 43 V. Operation in higher stability zones also has the advantage of aligned axial stability wells. That alignment maximizes the pseudopotential well depth for each higher stability zone, making them more than an order of magnitude greater than the standard ∼0.2 V well of a sine filter operating in the first stability zone at unit resolution. Increased pseudopotential well depth correlates with better ion transmission and sensitivity. Our theoretical examination suggests that the digital mass filter can obtain both high resolution and high sensitivity with essentially unlimited mass range.

Purpose Additive manufacturing (AM) of thermoplastic polymers for powder bed fusion processes typically requires each layer to be fused before the next can be deposited. The purpose of this paper is to present a volumetric AM method in the form of deeply penetrating radio frequency (RF) radiation to improve the speed of the process and the mechanical properties of the polymer parts. Design/methodology/approach The focus of this study was to demonstrate the volumetric fusion of composite mixtures containing polyamide (nylon) 12 and graphite powders using RF radiation as the sole energy source to establish the feasibility of a volumetric AM process for thermoplastic polymers. Impedance spectroscopy was used to measure the dielectric properties of the mixtures as a function of increasing graphite content and identify the percolation limit. The mixtures were then tested in a parallel plate electrode chamber connected to an RF generator to measure the heating effectiveness of different graphite concentrations. During the experiments, the surface temperature of the doped mixtures was monitored. Findings Nylon 12 mixtures containing between 10% and 60% graphite by weight were created, and the loss tangent reached a maximum of 35%. Selective RF heating was shown through the formation of fused composite parts within the powder beds. Originality/value The feasibility of a novel volumetric AM process for thermoplastic polymers was demonstrated in this study, in which RF radiation was used to achieve fusion in graphite-doped nylon powders.

The biology of human granzymes remains enigmatic in part due to our inability to probe their functions outside of in vitro assays or animal models with divergent granzyme species. We hypothesize that the biology of human granzymes could be better elaborated with a translational imaging technology to reveal the contexts in which granzymes are secreted and biochemically active in vivo. Here, we advance toward this goal by engineering a Granzyme targeting Restricted Interaction Peptide specific to family member B (GRIP B) to measure secreted granzyme B (GZMB) biochemistry with positron emission tomography. A proteolytic cleavage of 64Cu-labeled GRIP B liberates a radiolabeled form of Temporin L, which sequesters the radioisotope by binding to adjacent phospholipid bilayers. Thus, at extended time points postinjection (i.e., hours, not seconds), tissue biodistribution of the radioisotope in vivo reflects relative units of the GZMB activity. As a proof of concept, we show in three syngeneic mouse cancer models that 64Cu-GRIP B detects GZMB from T cells activated with immune checkpoint inhibitors (CPI). Remarkably, the radiotracer detects the proteolysis within tumors but also in lymphoid tissue, where immune cells are activated by a systemic CPI. Control experiments with an uncleavable analogue of 64Cu-GRIP B and tumor imaging studies in germline GZMB knockout mice were applied to show that 64Cu-GRIP B is specific for GZMB proteolysis. Furthermore, we explored a potential noncytotoxic function for GZMB by applying 64Cu-GRIP B to a model of pulmonary inflammation. In summary, we demonstrate that granzyme biochemistry can be assessed in vivo using an imaging modality that can be scaled vertically into human subjects., A protease-activated chemosensor for in vivo molecular imaging enables the detection of activated immune cells by specifically harnessing granzyme B catalysis.

The aim of this review is to explore how metabolic changes induced by diets high in saturated fat (HFD) affect nucleus accumbens (NAc) dopamine neurotransmission and food intake, and to explore how stress and inflammation influence this process. Recent evidence linked diet-induced obesity and HFD with reduced dopamine release and reuptake. Altered dopamine neurotransmission could disrupt satiety circuits between NAc dopamine terminals and projections to the hypothalamus. The NAc directs learning and motivated behaviours based on homeostatic needs and psychological states. Therefore, impaired dopaminergic responses to palatable food could contribute to weight gain by disrupting responses to food cues or stress, which impacts type and quantity of food consumed. Specifically, saturated fat promotes neuronal resistance to anorectic hormones and activation of immune cells that release proinflammatory cytokines. Insulin has been shown to regulate dopamine neurotransmission by enhancing satiety, but less is known about effects of diet-induced stress. Therefore, changes to dopamine signalling due to HFD warrant further examination to characterise crosstalk of cytokines with endocrine and neurotransmitter signals. A HFD promotes a proinflammatory environment that may disrupt neuronal endocrine function and dopamine signalling that could be exacerbated by the hypothalamic–pituitary–adrenal and κ-opioid receptor stress systems. Together, these adaptive changes may dysregulate eating by changing NAc dopamine during hedonic versus homeostatic food intake. This could drive palatable food cravings during energy restriction and hinder weight loss. Understanding links between HFD and dopamine neurotransmission will inform treatment strategies for diet-induced obesity and identify molecular candidates for targeted therapeutics.

We present the long-term retention and separation data reproducibility between 2013 and 2020 for analytical scale (4.6 mm internal diameter) superficially porous particle (SPP) columns. The retention factor for a small molecule—naphthalene—separated on 60 randomized manufactured lots, resulted in a 1.04% RSD; for a large molecule—bovine ribonuclease—separated on 31 randomized lots, 1.16% RSD. The peak shapes for three lots within this 7-year period were overlaid to visualize the chromatographic profile reproducibility. Naphthalene’s tailing factor had a % RSD of 4.42 and bovine ribonuclease’s peak width had an % RSD of 2.94; these metrics are sensitive to variability of the total error contribution of the column and system. A small and a large molecule application demonstrated reproducibility using three SPP manufactured lots packed in a narrow bore 2.1 mm i.d. analytical scale format. 15 peaks of the small molecule study resulted with a retention time %RSD reproducibility of ≤ 0.32, and for the large molecule study ≤ 1.12. The information in this study and the detailed discussion of the variability associated to different separation metrics is critical for industries that use HPLC and must adhere to stringent regulatory specifications, e.g., pharmaceutical, food and beverage industry.

The role of small secreted peptides in plant defense responses to viruses has seldom been investigated. Here, we report a role for potato (Solanum tuberosum) PIP1, a gene predicted to encode a member of the pathogen-associated molecular pattern (PAMP)-induced peptide (PIP) family, in the response of potato to Potato virus Y (PVY) infection. We show that exogenous application of synthetic StPIP1 to potato leaves and nodes increased the production of reactive oxygen species and the expression of plant defense-related genes, revealing that StPIP1 triggers early defense responses. In support of this hypothesis, transgenic potato plants that constitutively overexpress StPIP1 had higher levels of leaf callose deposition and, based on measurements of viral RNA titers, were less susceptible to infection by a compatible PVY strain. Interestingly, systemic infection of StPIP1-overexpressing lines with PVY resulted in clear rugose mosaic symptoms that were absent or very mild in infected non-transgenic plants. A transcriptomics analysis revealed that marker genes associated with both pattern-triggered immunity and effector-triggered immunity were induced in infected StPIP1 overexpressors but not in non-transgenic plants. Together, our results reveal a role for StPIP1 in eliciting plant defense responses and in regulating plant antiviral immunity.

In the pursuit to enable the rapid charging of lithium-ion batteries, lithium plating at the anode poses one of the most significant challenges. Additionally, the heat generation that accompanies high rate battery operation in conjunction with non-uniform cooling and localized heating at tabs is known to result in thermal inhomogeneity. Such thermal anomalies in the absence of proper thermal management can instigate accelerated degradation in the cell. In this work, a physics-based interrogation of the link between thermal gradient induced inhomogeneity and lithium plating during charging is presented. The relative importance of in-plane vs. through-plane (inter-electrode) thermal gradients to charging performance and cell degradation is necessary to intelligently design packaging and cooling systems for large-format cells. While in-plane thermal gradients strongly influence active material utilization, the lithium plating severity was found to be very similar to an isothermal case at the same mean temperature. By contrast, interelectrode thermal gradients cause a shifting on the solid phase potential at each electrode during charging, related to the increase or decrease in overpotential due to local temperature variation. When the cathode temperature exceeds the anode temperature, lithium plating is exacerbated, and accelerated degradation occurs.

Aquaporins (AQPs) are a ubiquitous family of transmembrane water channel proteins. A subgroup of AQP water channels also facilitates transmembrane diffusion of small, polar solutes. A constriction within the pore, the aromatic/arginine (ar/R) selectivity filter, is thought to control solute permeability: previous studies on single representative water channel proteins suggest narrow channels conduct water, whilst wider channels permit passage of solutes. To assess this model of selectivity, we used mutagenesis, permeability measurements and in silico comparisons of water-specific as well as glycerol-permeable human AQPs. Our studies show that single amino acid substitutions in the selectivity filters of AQP1, AQP4 and AQP3 differentially affect glycerol and urea permeability in an AQP-specific manner. Comparison between in silico-calculated channel cross-sectional areas and in vitro permeability measurements suggests that selectivity filter cross-sectional area predicts urea but not glycerol permeability. Our data show that substrate discrimination in water channels depends on a complex interplay between the solute, pore size, and polarity, and that using single water channel proteins as representative models has led to an underestimation of this complexity.

In this study, we report on the synthesis and evaluation of a sodium, magnesium, and aluminum (Na-Mg-Al) layered double hydroxide (LDH) based sorbent for hydrogen chloride (HCl) removal at concentrations found in lignocellulosic biomass derived syngas. The LDH was synthesized by a spontaneous self-assembly method and further calcined at 700 °C to produce a mixed metal oxide sorbent that we evaluated in the hot gas cleanup of hydrogen chloride at 100 parts per million. The performance of this sorbent was evaluated in a fixed bed reactor from 400 to 600 °C against that of a commercial magnesium and aluminum LDH (ComLDH) material that does not contain sodium in the matrix as well as two other commercial sorbents, sodium carbonate (Na2CO3) and sodium aluminate (NaAlO2). Our Na-Mg-Al LDH is thermally stable in the hot gas cleanup temperature range. During fixed bed experiments, our calcined LDH mixed metal oxide was effective in reducing hydrogen chloride’s concentration below the breakthrough concentration of 1 ppm from 400 to 600 °C for more than 14 h. The better performance of our calcined LDH compared to calcined commercial LDH supported our hypothesis that sodium incorporation in the LDH matrix enhances HCl sorption. Based on comparison against the commercial Na-based sorbents, the following rankings by temperature observed: LDH > NaAlO2 > Na2CO3 at 400 °C; LDH = NaAlO2 > Na2CO3 at 500 °C; and Na2CO3 = NaAlO2 = LDH at 600 °C.

This review provides an examination of the marsh spot disease in beans and the roles played by its causal factor, manganese (Mn) deficiency. The discovery of the marsh spot disease, its relation with Mn deficiency, and how it can be treated are discussed. Mn serves as a cofactor and a catalyst in various metabolic processes in different cell compartments, such as the oxygen-evolving complex of photosystem II (PSII) or reactive oxygen species scavenging. Some major quantitative trait loci (QTL) and putative candidate genes associated with Mn content in plants, especially in plant seeds, have been identified. Marsh spot disease in cranberry common bean is controlled by several major genes with significant additive and epistatic effects. They provide valuable clues for QTL candidate gene prediction and an improved understanding of the genetic mechanisms responsible for marsh spot resistance in plants.

Metallic lithium deposition on graphite anodes is a critical degradation mode in lithium-ion batteries, which limits safety and fast charge capability. A conclusive strategy to mitigate lithium deposition under fast charging yet remains elusive. In this work, we examine the role of electrode microstructure in mitigating lithium plating behavior under various operating conditions, including fast charging. The multilength scale characteristics of the electrode microstructure lead to a complex interaction of transport and kinetic limitations that significantly governs the cell performance and the occurrence of Li plating. We demonstrate, based on a comprehensive mesoscale analysis, that the performance and degradation can be significantly modulated via systematic design improvements at the hierarchy of length scales. It is found that the improvement in kinetic and transport characteristics achievable at disparate scales can dramatically affect Li plating propensity.

A wide variety of steroid metabolites synthesized by eukaryotes are all ultimately catabolized by bacteria; while generally saprophytic, pathogenic Mycobacteria have repurposed these pathways to utilize host intracellular cholesterol pools. Steroid degradation is complex, but a recurring theme is that cycles of β-oxidation are used to iteratively remove acetyl- or propanoyl-CoA groups. These β-oxidation cycles are initiated by the FAD-dependent oxidation of acyl groups, catalyzed by acyl-CoA dehydrogenases (ACADs). We show here that the

Background/objective The relationship between postprandial glycaemic responses and cognitive performance, mood and satiety are inconsistent. The objective of this study is to compare the effects of different glycaemic responses, induced by beverages with different glycaemic index (GI) (sucrose and isomaltulose), and a non-glycaemic control (sucralose), on cognition, mood and satiety. Subjects/methods In this double-blinded, randomised crossover trial, healthy adults (n = 55) received sucrose (GI 65), isomaltulose (GI 32) and sucralose (non-caloric negative control) drinks on separate occasions. The Complex Figure test, the Word Recall test, Trail Making Test Part B and the Stroop test were administered 60 min after beverages ingestion. Mood and satiety were tested along with cognitive performance. Results Comparing between isomaltulose and sucrose, there were no significant differences in the mean (95% CI) for the following: Complex Figure: immediate recall -0.6 (-1.7, 0.5), delayed recall -0.8 (-1.9, 0.3); Word recall: immediate recall 0.2 (-0.7, 1.1), delayed recall 0.5 (-0.4, 1.4); Trail Making: completing time -2.4 (-7.5, 2.7) s; Stroop: time used for correct congruent responses -9 (-31, 14) ms and correct incongruent responses -18 (-42, 6) ms. No differences among beverages were found in the mood and satiety scores with exception that participants felt more energetic 60 min after isomaltulose ingestion (p = 0.028 for difference with sucrose) and hungrier 30 min after isomaltulose ingestion (p = 0.036 for difference with sucrose; p = 0.022 for difference with sucralose). Conclusion Under these study conditions there is no convincing evidence for an effect of glycaemic response on cognitive performance, mood or satiety.

Hydrogen atom transfer from metal hydrides to alkenes appears to underlie widely used catalytic methods – the mechanistic implications are fascinating., Hydrogen atom transfer from a metal hydride (MHAT) has emerged as a powerful, if puzzling, technique in chemical synthesis. In catalytic MHAT reactions, earth-abundant metal complexes generate stabilized and unstabilized carbon-centered radicals from alkenes of various substitution patterns with robust chemoselectivity. This perspective combines organic and inorganic perspectives to outline challenges and opportunities, and to propose working models to assist further developments. We attempt to demystify the putative intermediates, the basic elementary steps, and the energetic implications, especially for cage pair formation, collapse and separation. Distinctions between catalysts with strong-field (SF) and weak-field (WF) ligand environments may explain some differences in reactivity and selectivity, and provide an organizing principle for kinetics that transcends the typical thermodynamic analysis. This blueprint should aid practitioners who hope to enter and expand this exciting area of chemistry.

Decomposing foliar litter organic matter (LOM) in forested watersheds is an essential terrestrial source of dissolved organic matter (DOM) to aquatic ecosystems. To evaluate the relationship of chemical composition between LOM and DOM, we collected freshly fallen leaves of white oak (Quercus alba) and conducted an 80-week field decomposition experiment along a small elevation gradient in a floodplain within the Congaree National Park, South Carolina. Foliar litters were collected for water extraction and instrumental analyses using pyrolysis GC/MS, UV/VIS and fluorescence spectrophotometry. Factor analyses of pyrolysates showed that fresh LOM was mainly composed of lignin compounds (LgC), phenolic compounds (PhC), and carbohydrate (Carb), and the change in composition was relatively small throughout the decomposition process. In contrast, there were two distinct chemical compositions of DOM in early (between 0–8 weeks) and late phases (between 16–80 weeks). The early phase had a higher percentage of PhC, but the late phase contained higher percentages of Carb, unsaturated hydrocarbon (UnSaH), aromatic hydrocarbon (ArH), and nitrogen-containing compounds (Ntg). The fluorescence emission-excitation matrix showed there was an increasing trend in humic and fulvic-like fractions in DOM over time, matching well with increases of UnSaH and ArH fractions from the results of pyrolysis GC/MS. Our study illustrated that the changes of chemical components in LOM and water extractable DOM were not parallel during the decomposition process and the degradation of lignin and phenolic compounds was one of the controlling factors on the production of DOM.

A hydrophilic ligand that contains only C, H, O, and N substituents and uses a 6,6′-bis(1H-1,2,3-triazol-4-yl)-2,2′-bipyridine (BTzBP) structural core has been synthesized. The effect of adding water-soluble groups onto extractant ligands has been extensively studied to facilitate the efficient partitioning of 4f and transuranic 5f elements for the treatment of spent nuclear fuel. Soft, N-donor ligands exhibit greater binding affinities for the trivalent actinides over the trivalent lanthanides, making BTzBP ligands an ideal candidate in the search for extractants to be used on an industrial scale. To date, hydrophobic BTzBPs have been shown to exhibit physical and chemical properties that might be conducive to nuclear waste processing conditions. However, hydrophilic BTzBPs have yet to be reported. Herein, we show the synthesis of a hydrophilic BTzBP ligand featuring cationic water solubilizing groups attached to the bipyridal rings.

Shipboard measurements of offshore oil and gas facilities were conducted in the Gulf of Mexico in February 2018. Species measured at 1 s include methane, ethane, carbon-13 (13C) and deuterium (D) i...

The Northern Niagaran Pinnacle Reef Trend has more than 800 identified Silurian-aged pinnacle reefs in the Michigan Basin. The reefs have a long history of oil and gas production, gas storage, and more recently successful CO2-enhanced oil recovery (EOR) operations. These fields provide an excellent opportunity to evaluate the geologic variability in complex carbonate reservoirs and its impact on carbon capture, utilization, and storage feasibility. A comprehensive database was built that identifies reefs and links key reservoir attributes to each field. Novel methodologies and techniques were developed to analyze hundreds of fields for CO2 storage and EOR options. This included a regional static earth model to compute volumetric-based resource estimates, fluid substitution to estimate storage resources from oil and gas production, and proximity analysis–based weighting of reefs to predict reservoir performance metrics and estimate EOR. Lastly, an enhanced storage scenario of maximizing a reef’s storage potential at the end of a CO2-EOR life cycle was evaluated.

To fight the SARS-CoV-2 pandemic, much effort has been directed toward drug repurposing, testing investigational and approved drugs against several viral or human proteins in vitro. Here we investigate the impact of colloidal aggregation, a common artifact in early drug discovery, in these repurposing screens. We selected 56 drugs reported to be active in biochemical assays and tested them for aggregation by both dynamic light scattering and by enzyme counter screening with and without detergent; seventeen of these drugs formed colloids at concentrations similar to their literature reported IC50s. To investigate the occurrence of colloidal aggregators more generally in repurposing libraries, we further selected 15 drugs that had physical properties resembling known aggregators from a common repurposing library, and found that 6 of these aggregated at micromolar concentrations. An attraction of repurposing is that drugs active on one target are considered de-risked on another. This study suggests not only that many of the drugs repurposed for SARS-CoV-2 in biochemical assays are artifacts, but that, more generally, when screened at relevant concentrations, drugs can act artifactually via colloidal aggregation. Understanding the role of aggregation, and detecting its effects rapidly, will allow the community to focus on those drugs and leads that genuinely have potential for treating COVID-19.Abstract FigureTable of Contents Graphic

Understanding how oxidatively damaged RNA is handled intracellularly is of relevance due to the link between oxidized RNA and the progression/development of some diseases as well as aging. Among the ribonucleases responsible for the decay of modified (chemically or naturally) RNA is the exonuclease Xrn-1, a processive enzyme that catalyzes the hydrolysis of 5′-phosphorylated RNA in a 5′→3′ direction. We set out to explore the reactivity of this exonuclease towards oligonucleotides (ONs, 20-nt to 30-nt long) of RNA containing 8-oxo-7,8-dihydroguanosine (8-oxoG), obtained via solid-phase synthesis. The results show that Xrn-1 stalled at sites containing 8-oxoG, evidenced by the presence of a slower moving band (via electrophoretic analyses) than that observed for the canonical analogue. The observed fragment(s) were characterized via PAGE and MALDI-TOF to confirm that the oligonucleotide fragment(s) contained a 5′-phosphorylated 8-oxoG. Furthermore, the yields for this stalling varied from app. 5–30% with 8-oxoG located at different positions and in different sequences. To gain a better understanding of the decreased nuclease efficiency, we probed: 1) H-bonding and spatial constraints; 2) anti-syn conformational changes; 3) concentration of divalent cation; and 4) secondary structure. This was carried out by introducing methylated or brominated purines (m1G, m6,6A, or 8-BrG), probing varying [Mg2+], and using circular dichroism (CD) to explore the formation of structured RNA. It was determined that spatial constraints imposed by conformational changes around the glycosidic bond may be partially responsible for stalling, however, the results do not fully explain some of the observed higher stalling yields. We hypothesize that altered π-π stacking along with induced H-bonding interactions between 8-oxoG and residues within the binding site may also play a role in the decreased Xrn-1 efficiency. Overall, these observations suggest that other factors, yet to be discovered/established, are likely to contribute to the decay of oxidized RNA. In addition, Xrn-1 degraded RNA containing m1G, and stalled mildly at sites where it encountered m6,6A, or 8-BrG, which is of particular interest given that the former two are naturally occurring modifications.

Resume— Les effets d'alimentations supplementees avec des ratios croissants decides gras polyinsatures n — 6/n — 3 sur la synthese de leucotrienes B dans la peau du chien et les neutrophiles sont presentes. Trente chien Beagles ont recu pendant 2 mois avec une alimentation ayant un ratio n —6/n —3 de 28:1. Des aliments experimentaux contenant des ratios de 5:1, 10:1, 25:1, 50:1 et 100:1 (six chiens par groupe) ont ete administres ensuite pendant 12 semaines. A la fin des deux mois d'alimentation temoin et au bout de 6 et 12 semaines d'alimentation experimentale, les concentrations de LTB4 et LTB5 dans la peau et les neutrophiles ont ete determinees. Les neutrophiles de chiens ayant mange des aliments de ratio 5:1 et 10:1 ont synthetise 30–33 pourcent moins de LTB4 (P

The CST complex (CTC1-STN1-TEN1) has been shown to inhibit telomerase extension of the G-strand of telomeres and facilitate the switch to C-strand synthesis by DNA polymerase alpha-primase (pol α-primase). Recently the structure of human CST was solved by cryo-EM, allowing the design of mutant proteins defective in telomeric ssDNA binding and prompting the reexamination of CST inhibition of telomerase. The previous proposal that human CST inhibits telomerase by sequestration of the DNA primer was tested with a series of DNA-binding mutants of CST and modeled by a competitive binding simulation. The DNA-binding mutants had substantially reduced ability to inhibit telomerase, as predicted from their reduced affinity for telomeric DNA. These results provide strong support for the previous primer sequestration model. We then tested whether addition of CST to an ongoing processive telomerase reaction would terminate DNA extension. Pulse-chase telomerase reactions with addition of either wild-type CST or DNA-binding mutants showed that CST has no detectable ability to terminate ongoing telomerase extension in vitro. The same lack of inhibition was observed with or without pol α-primase bound to CST. These results suggest how the switch from telomerase extension to C-strand synthesis may occur., Graphical Abstract Graphical AbstractHuman CST inhibits telomerase engagement but not elongation.

Angiogenesis is critical for successful fracture healing. Age-related alterations in endothelial cells (ECs) may cause impaired bone healing. Therefore, examining therapeutic treatments to improve angiogenesis in aging may enhance bone healing. Sirtuin 1 (SIRT1) is highly expressed in ECs and its activation is known to counteract aging. Here, we examined the effects of SRT1720 treatment (SIRT1 activator) on the growth and function of bone marrow and lung ECs (BMECs and LECs, respectively), derived from young (3–4 month) and old (20–24 month) mice. While aging did not alter EC proliferation, treatment with SRT1720 significantly increased proliferation of all LECs. However, SRT1720 only increased proliferation of old female BMECs. Vessel-like tube assays showed similar vessel-like structures between young and old LECs and BMECs from both male and female mice. SRT1720 significantly improved vessel-like structures in all LECs. No age, sex, or treatment differences were found in migration related parameters of LECs. In males, old BMECs had greater migration rates than young BMECs, whereas in females, old BMECs had lower migration rates than young BMECs. Collectively, our data suggest that treatment with SRT1720 appears to enhance the angiogenic potential of LECs irrespective of age or sex. However, its role in BMECs is sex- and age-dependent.

Redox reactions that take place in enzymes and on the surfaces of heterogeneous catalysts often require active sites that contain multiple metals. By contrast, there are very few homogeneous catalysts with multinuclear active sites, and the field of organometallic chemistry continues to be dominated by the study of single metal systems. Multinuclear catalysts have the potential to display unique properties owing to their ability to cooperatively engage substrates. Furthermore, direct metal-to-metal covalent bonding can give rise to new electronic configurations that dramatically impact substrate binding and reactivity. In order to effectively capitalize on these features, it is necessary to consider strategies to avoid the dissociation of fragile metal-metal bonds in the course of a catalytic cycle. This Account describes one approach to accomplishing this goal using binucleating redox-active ligands.In 2006, Chirik showed that pyridine-diimines (PDI) have sufficiently low-lying π* levels that they can be redox-noninnocent in low-valent iron complexes. Extending this concept, we investigated a series of dinickel complexes supported by naphthyridine-diimine (NDI) ligands. These complexes can promote a broad range of two-electron redox processes in which the NDI ligand manages electron equivalents while the metals remain in a Ni(I)-Ni(I) state.Using (NDI)Ni2 catalysts, we have uncovered cases where having two metals in the active site addresses a problem in catalysis that had not been adequately solved using single-metal systems. For example, mononickel complexes are capable of stoichiometrically dimerizing aryl azides to form azoarenes but do not turn over due to strong product inhibition. By contrast, dinickel complexes are effective catalysts for this reaction and avoid this thermodynamic sink by binding to azoarenes in their higher-energy cis form.Dinickel complexes can also activate strong bonds through the cooperative action of both metals. Norbornadiene has a ring-strain energy that is similar to that of cyclopropane but is not prone to undergoing C-C oxidative addition with monometallic complexes. Using an (NDI)Ni2 complex, norbornadiene undergoes rapid ring opening by the oxidative addition of the vinyl and bridgehead carbons. An inspection of the resulting metallacycle reveals that it is stabilized through a network of secondary Ni-π interactions. This reactivity enabled the development of a catalytic carbonylative rearrangement to form fused bicyclic dienones.These vignettes and others described in this Account highlight some of the implications of metal-metal bonding in promoting a challenging step in a catalytic cycle or adjusting the thermodynamic landscape of key intermediates. Given that our studies have focused nearly exclusively on the (NDI)Ni2 system, we anticipate that many more such cases are left to be discovered as other transition-metal combinations and ligand classes are explored.

Additive manufacturing (AM) of high-performance composites has gained increasing interest over the last few years. Commercially available AM technologies often use thermoplastics as they are easy to process, i.e., to melt and re-solidify. However, thermosetting polymers generally achieve superior mechanical properties and thermostability. This study investigates reactive extrusion additive manufacturing (REAM) of a thermosetting polymer reinforced with carbon fibers. The process utilizes highly exothermic and fast curing resin/catalyst systems, eliminating the need for post-curing. The rheological properties of the liquid resin are first tuned for REAM using ~2wt.% fumed silica and ~10vol.% milled carbon fibers. Then, a robotic arm is used to print the composite samples. The coupons’ longitudinal and transverse tensile properties are measured and correlated with the degree of cure, porosity, fiber length distribution, and fiber orientation distribution. The incorporation of milled carbon fibers, 50-200 m long, primarily affects the stiffness. Compared to neat polymer parts, carbon fiber reinforced composites are 51% stiffer and 8% stronger. In addition, polymeric crosslinking between part layers resulted in strong inter-layer bonding. Short fibers were also randomly oriented within parts due to the nozzle size and shape, resulting in nearly isotropic parts. The results presented here pave the road for fast and low-energy AM of high-performance composites.

The objective of the current study was to evaluate the color changes and lipid oxidation of ground chicken patties packaged in polyvinyl chloride (PVC) film, high-oxygen (HiOx)–modified atmospheric packaging (MAP; 80% oxygen + 20% carbon dioxide [CO2]), and carbon monoxide (CO)-MAP (0.4% CO + 19.6% CO2 + 80% nitrogen) and stored at 2°C. Surface color was measured using a HunterLab MiniScan spectrophotometer on days 0, 1, 2, and 4. Lipid oxidation, pH, and aerobic plate count were determined on days 0 and 4 of storage. Fatty acid profiles were determined on day 0 to characterize saturated and unsaturated fatty acids. Patties packaged in PVC had greater (P < 0.05) pH than HiOx-MAP and CO-MAP. Gas chromatography analysis indicated that ground chicken has 72.8% unsaturated fatty acids and 27.2% saturated fatty acids (based on total lipids and fatty acid methyl ester). The formation of carboxymyoglobin on ground chicken patty surface was confirmed by peaks at 420 and 570 nm, whereas oxymyoglobin had peaks at 410 and 580 nm. Instrumental color analysis indicated both HiOx-MAP and CO-MAP had greater (P < 0.05) redness (a* values) than PVC on day 4 of storage. Patties packaged in HiOx-MAP had greater (P < 0.05) chroma values than CO-MAP and PVC on day 4 of storage. Visual panelists noted less (P < 0.05) surface discoloration in CO-MAP than PVC and HiOx-MAP on day 4 of storage. Lipid oxidation was greater (P < 0.05) in PVC and HiOx-MAP than CO-MAP. CO inclusion at 0.4% level effectively inhibited lipid oxidation and stabilized surface redness during refrigerated storage of ground chicken.

In this study, we design, fabricate and characterize a novel unimorph-like structure in which a shape memory polymer (SMP) is actuated using a magneto-active elastomer (MAE), resulting in contactless and large deformation shape change. The constituent materials are: a two-part elastomer, iron oxide microparticles, and a rapid cure epoxy. Combining the elastomer with the iron oxide particles results in an MAE actuator, such that application of an external magnetic field induces dipoles in the iron oxide particles, resulting in the actuation of the MAE. The shape memory epoxy polymers are deformed at an elevated temperature and cooled to lock-in the temporary programmed shape. Subsequent exposure to elevated temperature can then recover a permanent geometry. In a first step, the layered unimorph structure is fabricated using casting techniques in which the SMP serves as the ‘passive’ substrate, and the MAE is cast on top of this substrate or joined post-processing using adhesives. To demonstrate shape change, the unimorph is heated beyond the transition temperature of the epoxy, and a magnetic field is applied to deform the structure. Upon cooling, the shape change is locked into place. This configuration serves as proof-of-concept that the proposed magnetically-responsive polymers are capable of eliciting the shape change of an SMP through bending in response to an external magnetic stimulus. After demonstrating feasibility, the next step is to use additive manufacturing (AM) to produce both the SMP and MAE by developing and tuning MAE and SMP resin formulations for printability. Printability is assessed by characterizing the viscosities, effective yield stress, gelation times, and extent of cure of the material formulations. The bending response of the layered structure is characterized as a function of magnetic field, material composition, geometric parameters, and AM process settings. The outcome of this research is to enable AM of smart materials and devices that can monitor and adapt to their environment. The medical device industry in particular stands to benefit from customizable devices that adapt their shape for particular patients, diseases, and/or stage of healing.

Genetic deletion of Phosphoinositide 3-kinase (PI3Kγ) in mice (PI3Kγ -/- ) results in increased cAMP levels and enhanced ventricular rate/contractility. We investigated whether PI3Kγ plays a role in cardiac contractility by altering intracellular calcium recycling. Caffeine treatment of adult cardiomyocytes from PI3Kγ -/- mice showed significantly reduced calcium reuptake by sarcoendoplasmic reticulum (SR) indicating that PI3Kγ locally regulates SR function. This resulted in elevated levels of intracellular calcium for prolonged period following caffeine. Our findings show that delayed re-uptake of calcium was caused by changes in phosphorylation of phospholamban (PLN), a major regulator of SR calcium reuptake. PI3Kγ -/- cardiomyocytes showed significantly reduced PLN phosphorylation due to increase in PLN-associated protein phosphatase (PP) activity as reflected by decreased demethylated-PP2A. Abrogation of PLN phosphorylation in the PI3Kγ -/- cardiomyocytes shows that the loss in the steady-state phosphorylation of PLN leads to increased inhibition of SERCA. This inhibition is reflected by the slow reuptake of calcium by the SR in the PI3Kγ -/- cardiomyocytes. Concomitantly, significant interaction was observed between SERCA and PLN in the PI3Kγ -/- hearts compared to the controls. Consistently, the altered calcium regulation in the cardiomyocytes of PI3Kγ -/- can be restored by inhibition of PP by okadaic acid. Unexpectedly, cardiomyocyte-specific overexpression of kinase-dead PI3Kγ (PI3Kγ inact ) in the global PI3Kγ -/- cardiomyocytes normalized caffeine induced calcium reuptake, restored PLN phosphorylation, and decreased PLN-associated PP activity reflected by increased demethylated-PP2A. These studies bring-to-fore an unrecognized kinase-independent regulation of PLN by PI3Kγ through PP2A with implications in deleterious cardiac remodeling as PI3Kγ is significantly upregulated following cardiac stress.

Aquaporins and aquaglyceroporins are membrane channels that allow the permeation of water and certain other small solutes across the cell membrane, or in the case of AQP6, AQP11 and AQP12A, intracellular membranes, such as vesicles and the endoplasmic reticulum membrane [16]. Since the isolation and cloning of the first aquaporin (AQP1) [20], 12 additional mammalian members of the family have been identified, although little is known about the functional properties of one of these (AQP12A; Q8IXF9) and it is thus not tabulated. The other 12 aquaporins can be broadly divided into three families: orthodox aquaporins (AQP0,-1,-2,-4,-5, -6 and -8) permeable mainly to water, but for some additional solutes [4]; aquaglyceroporins (AQP3,-7 -9 and -10), additionally permeable to glycerol and for some isoforms urea [14], and superaquaporins (AQP11 and 12) located within cells [12]. Some aquaporins also conduct ammonia and/or H2O2 giving rise to the terms 'ammoniaporins' ('aquaammoniaporins') and 'peroxiporins', respectively. Aquaporins are impermeable to protons and other inorganic and organic cations, with the possible exception of AQP1, although this is controversial [14]. One or more members of this family of proteins have been found to be expressed in almost all tissues of the body [reviewed in Yang (2017) [26]]. AQPs are involved in numerous processes that include systemic water homeostasis, adipocyte metabolism, brain oedema, cell migration and fluid secretion by epithelia. Loss of function mutations of some human AQPs, or their disruption by autoantibodies further underscore their importance [reviewed by Verkman et al. (2014) [23], Kitchen et al. (2105) [14]]. Functional AQPs exist as homotetramers that are the water conducting units wherein individual AQP subunits (each a protomer) have six TM helices and two half helices that constitute a seventh 'pseudotransmembrane domain' that surrounds a narrow water conducting channel [16]. In addition to the four pores contributed by the protomers, an additional hydrophobic pore exists within the center of the complex [16] that may mediate the transport through AQP1. Although numerous small molecule inhibitors of aquaporins, particularly APQ1, have been reported primarily from Xenopus oocyte swelling assays, the activity of most has subsequently been disputed upon retesting using assays of water transport that are less prone to various artifacts [5] and they are therefore excluded from the tables [see Tradtrantip et al. (2017) [22] for a review].