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Elution chromatography can be carried out in either isocratic or gradient modes. In isocratic elution, the mobile-phase composition is held constant during separation of the sample, e.g. 60% v acetonitrile–water. In gradient elution, the mobile-phase composition will be varied during sample separation, e.g. changing from 0 to 100% v acetonitrile–water. Gradient elution requires special chromatographic equipment, as well as somewhat greater care on the part of the operator, but it has important advantages for many separations. Thus, in isocratic elution (Figure 1a), sample peaks tend to “bunch up” at the beginning of the chromatogram (often with decreased resolution) and to broaden at the end of the chromatogram (with reduced detection sensitivity). Gradient elution (Figure 1b), on the other hand, provides a more even spacing of peaks, similar widths throughout the chromatogram, and often a shorter run time. For these and other reasons, gradient elution is preferred for the separation of many samples.

The uptake of calcium into and extrusion of calcium from the mitochondrial matrix is a fundamental biological process that has critical effects on cellular metabolism, signaling, and survival. Disruption of mitochondrial calcium (mCa2+) cycling is implicated in numerous acquired diseases such as heart failure, stroke, neurodegeneration, diabetes, and cancer and is genetically linked to several inherited neuromuscular disorders. Understanding the mechanisms responsible for mCa2+ exchange therefore holds great promise for the treatment of these diseases. The past decade has seen the genetic identification of many of the key proteins that mediate mitochondrial calcium uptake and efflux. Here, we present an overview of the phenomenon of mCa2+ transport and a comprehensive examination of the molecular machinery that mediates calcium flux across the inner mitochondrial membrane: the mitochondrial uniporter complex (consisting of MCU, EMRE, MICU1, MICU2, MICU3, MCUB, and MCUR1), NCLX, LETM1, the mitochondrial ryanodine receptor, and the mitochondrial permeability transition pore. We then consider the physiological implications of mCa2+ flux and evaluate how alterations in mCa2+ homeostasis contribute to human disease. This review concludes by highlighting opportunities and challenges for therapeutic intervention in pathologies characterized by aberrant mCa2+ handling and by summarizing critical unanswered questions regarding the biology of mCa2+ flux.

The study of serpentinites and ophicarbonates from ophiolitic terrains provides a three-dimensional perspective on the hydration and carbonation processes affecting modern oceanic lithosphere. The Chenaillet ophiolite (western Alps) is interpreted as a fragment of an oceanic core complex that resembles a modern slow spreading center, and it was weakly affected by Alpine metamorphism. Ophicarbonates from the Chenaillet ophiolite were targeted in this study for in situ analysis by Secondary Ion Mass Spectrometry (SIMS) of oxygen and carbon isotopes in serpentine, calcite, dolomite and magnetite. The high spatial resolution of SIMS allowed us to target different serpentine, carbonate and magnetite generations intergrown at scales ≤ 50 μm, and reveal systematic zoning in δ18O with a range of 5.8‰ in serpentine (from 3.0 to 8.8‰, V-SMOW), 21.2‰ in carbonate (9.4 to 30.6‰), and 5.6‰ in magnetite (–5.0 to –10.6‰). Coupled analysis of oxygen isotopes in seven different touching-pairs of co-crystallized serpentine+carbonate and serpentine+magnetite provides independent constraints on both the temperatures and δ18O(water) values during serpentinization and carbonation responsible for the formation of the Chenaillet ophicarbonates. The new stable isotope data and thermometric estimates can be directly linked to textural and petrographic observations. These new results identify at least four different stages of hydrothermal alteration in the Chenaillet ophicarbonates: (1) peridotite hydration during seafloor exhumation at temperatures down to 200-130 °C and water δ18O values varying from 5 to 2‰, as documented by serpentine+magnetite in mesh textures; (2) carbonation during exhumation near the seafloor at temperatures as low as 10 °C assuming water δ18O values of –1‰, as documented by the highest oxygen isotope ratios in texturally older calcite; (3) serpentinization and carbonation at temperatures up to 240 °C and water δ18O values of 2-3‰, as documented by serpentine+magnetite in veins crosscutting mesh textures (T = 192±66 °C, δ18O(water) = 2±1‰, 2 standard deviation), serpentine+magnetite (T = 182±32 °C, δ18O(water) = 2±1‰) and serpentine+dolomite (T = 243±79 °C, δ18O(water) = 3±2‰) in recrystallized hourglass domains within serpentinite clasts, serpentine+dolomite (T = 229±50 °C, δ18O(water) = 3±1‰) and serpentine+calcite (T = 208±40 °C, δ18O(water) = 2±1‰) within the fine-grained calcite matrix surrounding serpentinite clasts; (4) late stage carbonation at temperatures down to 70-40 °C assuming water δ18O values of 3 to –1‰, as documented by the highest oxygen isotope ratios in a large calcite vein crosscutting both serpentinite clasts and fine-grained carbonate matrix. We suggest that the textural and isotopic observations are consistent with a protracted serpentinization and carbonation of the lithospheric mantle that started during progressive exhumation to the seafloor and continued due to interaction with hot and isotopically shifted seawater, which circulated at depth in the oceanic crust and was then discharged near the seafloor, similar to modern mid-ocean ridge venting systems.

Neuro-endocrine prostate cancer (NEPC) accounts for about 20% of lethal metastatic castration-resistant prostate cancer (CRPC). NEPC has the most aggressive biologic behavior of all prostate cancers and is associated with poor patient outcome. Effective treatment for NEPC is not available because NEPC exhibit distinct cell-surface expression profiles compared to other types of prostate cancer. Recently, the carcinoembryonic antigen-related cell adhesion molecule 5 (CEACAM5) (known as CEA or CD66e) was suggested to be a specific surface protein marker for NEPC. Therefore, we identified a new, fully-human anti-CEACAM5 monoclonal antibody, 1G9, which bound to the most proximal membrane domains, A3 and B3, of CEACAM5 with high affinity and specificity. It shows no off-target binding to other CEACAM family members, membrane distal domains of CEACAM5, or 5800 human membrane proteins. IgG1 1G9 exhibited CEACAM5-specific ADCC activity toward CEACAM5-positive prostate cancer cells in vitro and in vivo. Chimeric antigen receptor T cells (CAR-T) based on scFv 1G9 induced specific and strong antitumor activity in a mouse model of prostate cancer. Our results suggest that IgG1 and CAR-T cells based on 1G9 are promising candidate therapeutics for CEACAM5-positive NEPC and other cancers.

Central adiposity is associated with greater sympathetic support of blood pressure. β-adrenergic receptors (β-AR) buffer sympathetically mediated vasoconstriction and β-AR-mediated vasodilation is attenuated in preclinical models of obesity. With this information, we hypothesized β-AR vasodilation would be lower in obese compared with normal weight adults. Because β-AR vasodilation in normal weight adults is limited by cyclooxygenase (COX) restraint of nitric oxide synthase (NOS), we further explored the contributions of COX and NOS to β-AR vasodilation in this cohort. Forearm blood flow (FBF, Doppler ultrasound) and mean arterial blood pressure (MAP, brachial arterial catheter) were measured and forearm vascular conductance (FVC) was calculated (FVC = FBF/MAP). The rise in FVC from baseline (ΔFVC) was quantified during graded brachial artery infusion of isoproterenol (Iso, 1–12 ng/100 g/min) in normal weight (n = 36) and adults with obesity (n = 22) (18–40 yr old). In a subset of participants, Iso-mediated vasodilation was examined before and during inhibition of NOS [N(G)-monomethyl-l-arginine (l-NMMA)], COX (ketorolac), and NOS + COX (l-NMMA + ketorolac). Iso-mediated increases in FVC did not differ between groups (P = 0.57). l-NMMA attenuated Iso-mediated ΔFVC in normal weight (P = 0.03) but not adults with obesity (P = 0.27). In normal weight adults, ketorolac increased Iso-mediated ΔFVC (P < 0.01) and this response was lost with concurrent l-NMMA (P = 0.67). In contrast, neither ketorolac (P = 0.81) nor ketorolac + l-NMMA (P = 0.40) altered Iso-mediated ΔFVC in adults with obesity. Despite shifts in COX and NOS, β-AR vasodilation is preserved in young adults with obesity. These data highlight the presence of a compensatory shift in microvascular control mechanisms in younger humans with obesity. NEW & NOTEWORTHY We examined β-adrenergic receptor-mediated vasodilation in skeletal muscle of humans with obesity and normal weight. Results show that despite shifts in the contribution of cyclooxygenase and nitric oxide synthase, β-adrenergic-mediated vasodilation is relatively preserved in young, otherwise healthy adults with obesity. These data highlight the presence of subclinical changes in microvascular control mechanisms early in the obesity process and suggest duration of obesity and/or the addition of primary aging may be necessary for overt dysfunction.

Liquid-solid or normal-phase chromatography is employed for the separation of non-ionic solutes by adsorption onto the surface of an inorganic oxide stationary phase and displacement by competition with the components of the mobile phase. It has particular advantages for compound-class separations and the separation of diasteromers and geometrical isomers soluble in organic solvents. Common stationary phases for high-performance liquid chromatography include silica and chemically-bonded silica phases with 3-cyanopropyl, 3-aminopropyl, and spacer bonded propanediol groups. The use of eluotropic series for mobile phase selection are described as well as general strategies for method development.

Significance Magnetic resonance imaging is hindered by inherently low sensitivity, which limits the method for the most part to observing water molecules in the body. Hyperpolarized molecules exhibit strongly enhanced MRI signals which opens the door for imaging low-concentration species in vivo. Biomolecules can be hyperpolarized and injected into a patient allowing for metabolism to be tracked in real time, greatly expanding the information available to the radiologist. Parahydrogen-induced polarization (PHIP) is a hyperpolarization method renowned for its low cost and accessibility, but is generally limited by low polarization levels, modest molecular concentrations, and contamination by polarization reagents. In this work we overcome these drawbacks in the production of PHIP-polarized [1-13C]fumarate, a biomarker of cell necrosis in metabolic 13C MRI., Hyperpolarized fumarate is a promising biosensor for carbon-13 magnetic resonance metabolic imaging. Such molecular imaging applications require nuclear hyperpolarization to attain sufficient signal strength. Dissolution dynamic nuclear polarization is the current state-of-the-art methodology for hyperpolarizing fumarate, but this is expensive and relatively slow. Alternatively, this important biomolecule can be hyperpolarized in a cheap and convenient manner using parahydrogen-induced polarization. However, this process requires a chemical reaction, and the resulting solutions are contaminated with the catalyst, unreacted reagents, and reaction side-product molecules, and are hence unsuitable for use in vivo. In this work we show that the hyperpolarized fumarate can be purified from these contaminants by acid precipitation as a pure solid, and later redissolved to a desired concentration in a clean aqueous solvent. Significant advances in the reaction conditions and reactor equipment allow for formation of hyperpolarized fumarate at 13C polarization levels of 30–45%.

For many years, lipid nanoparticles (LNPs) have been used as delivery vehicles for various payloads (especially various oligonucleotides and mRNA), finding numerous applications in drug and vaccine development. LNP stability and bilayer fluidity are determined by the identities and the amounts of the various lipids employed in the formulation and LNP efficacy is determined in large part by the lipid composition which usually contains a cationic lipid, a PEG‐lipid conjugate, cholesterol, and a zwitterionic helper phospholipid. Analytical methods developed for LNP characterization must be able to determine not only the identity and content of each individual lipid component (i.e., the parent lipids), but also the associated impurities and degradants. In this work, we describe an efficient and sensitive reversed‐phase chromatographic method with charged aerosol detection (CAD) suitable for this purpose. Sample preparation diluent and mobile phase pH conditions are critical and have been optimized for the lipids of interest. This method was validated for its linearity, accuracy, precision, and specificity for lipid analysis to support process and formulation development for new drugs and vaccines. This article is protected by copyright. All rights reserved

Guiding neuronal cell growth is desirable for neural tissue engineering but is very challenging. In this work, a self-assembling ultra-short surfactant-like peptide I3K which possesses positively charged lysine head groups, and hydrophobic isoleucine tails, was chosen to investigate its potential for guiding neuronal cell growth. The peptides were able to self-assemble into nanofibrous structures and interact strongly with silk fibroin (SF) scaffolds, providing a niche for neural cell attachment and proliferation. SF is an excellent biomaterial for tissue engineering. However neuronal cells, such as rat PC12 cells, showed poor attachment on pure regenerated SF (RSF) scaffold surfaces. Patterning of I3K peptide nanofibers on RSF surfaces significantly improved cellular attachment, cellular density, as well as morphology of PC12 cells. The live / dead assay confirmed that RSF and I3K have negligible cytotoxicity against PC12 cells. Atomic force microscopy (AFM) was used to image the topography and neurite formation of PC12 cells, where results revealed that self-assembled I3K nanofibers can support the formation of PC12 cell neurites. Immunolabelling also demonstrated that coating of I3K nanofibers onto the RSF surfaces not only increased the percentage of cells bearing neurites but also increased the average maximum neurite length. Therefore, the peptide I3K could be used as an alternative to poly-l-lysine for cell culture and tissue engineering applications. As micro-patterning of neural cells to guide neurite growth is important for developing nerve tissue engineering scaffolds, inkjet printing was used to pattern self-assembled I3K peptide nanofibers on RSF surfaces for directional control of PC12 cell growth. The results demonstrated that inkjet-printed peptide micro-patterns can effectively guide the cell alignment and organization on RSF scaffold surfaces, providing great potential for nerve regeneration applications.

Uranium (U) contamination in groundwater from geogenic sources affects water quality globally. Here, we use a multifaceted isotopic and geochemical approach to elucidate U sources and controls on g...

There is evidence that the cystic fibrosis transmembrane conductance regulator (CFTR) anion channel is highly expressed at the apical pole of ciliated cells in human bronchial epithelium (HBE), however recent studies have detected little CFTR mRNA in those cells. To understand this discrepancy we immunostained well differentiated primary HBE cells using CFTR antibodies. We confirmed apical immunofluorescence in ciliated cells and quantified the covariance of the fluorescence signals and that of an antibody against the ciliary marker centrin-2 using image cross-correlation spectroscopy (ICCS). Super-resolution stimulated emission depletion (STED) imaging localized the immunofluorescence in distinct clusters at the bases of the cilia. However, similar apical fluorescence was observed when the monoclonal CFTR antibodies 596, 528 and 769 were used to immunostain ciliated cells expressing F508del-CFTR, or cells lacking CFTR due to a Class I mutation. A BLAST search using the CFTR epitope identified a similar amino acid sequence in the ciliary protein rootletin X1. Its expression level correlated with the intensity of immunostaining by CFTR antibodies and it was detected by 596 antibody after transfection into CFBE cells. These results may explain the high apparent expression of CFTR in ciliated cells and reports of anomalous apical immunofluorescence in well differentiated cells that express F508del-CFTR.

Tellurium-rich (Te) adularia-sericite epithermal Au-Ag deposits are an important current and future source of precious and critical metals. However, the source and evolution of ore-forming fluids in these deposits are masked by traditional bulk analysis of quartz oxygen isotope ratios that homogenize fine-scale textures and growth zones. To advance understanding of the source of Te and precious metals, herein, we use petrographic and cathodoluminescence (CL) images of such textures and growth zones to guide high spatial resolution secondary ion mass spectroscopy (SIMS) oxygen isotope analyses (10 µm spot) and spatially correlated fluid inclusion microthermometric measurements on successive quartz bands in contemporary Te-rich and Te-poor adularia-sericite (-quartz) epithermal Au-Ag vein deposits in northeastern China. The results show that large positive oxygen isotope shifts from –7.1 to +7.7‰ in quartz rims are followed by precipitation of Au-Ag telluride minerals in the Te-rich deposit, whereas small oxygen isotope shifts of only 4‰ (–2.2 to +1.6‰) were detected in quartz associated with Au-Ag minerals in the Te-poor deposits. Moreover, fluid-inclusion homogenization temperatures are higher in comb quartz rims (avg. 266.4 to 277.5 °C) followed by Au-Ag telluride minerals than in previous stages (~250 °C) in the Te-rich deposit. The Te-poor deposit has a consistent temperature (~245 °C) in quartz that pre- and postdates Au-Ag minerals. Together, the coupled increase in oxygen isotope ratios and homogenization temperatures followed by precipitation of Au-Ag tellurides strongly supports that inputs of magmatic fluid containing Au, Ag, and Te into barren meteoric water-dominated flow systems are critical to the formation of Te-rich adularia-sericite epithermal Au-Ag deposits. In contrast, Te-poor adularia-sericite epithermal Au-Ag deposits show little or no oxygen isotope or fluid-inclusion evidence for inputs of magmatic fluid.

Oxygen vacancies and lattice oxygen on the surface of nanoparticles play crucial roles in the magnetism of transition metal oxides. A fundamental understanding of the interactions between the 2p or...

Background Erythrocyte transfusions are independently associated with acute kidney injury. Kidney injury may be consequent to the progressive hematologic changes that develop during storage. This study therefore tested the hypothesis that prolonged erythrocyte storage increases posttransfusion acute kidney injury. Methods The Informing Fresh versus Old Red Cell Management (INFORM) trial randomized 31,497 patients to receive either the freshest or oldest available matching erythrocyte units and showed comparable mortality with both. This a priori substudy compared the incidence of posttransfusion acute kidney injury in the randomized groups. Acute kidney injury was defined by the creatinine component of the Kidney Disease: Improving Global Outcomes criteria. Results The 14,461 patients included in this substudy received 40,077 erythrocyte units. For patients who received more than one unit, the mean age of the blood units was used as the exposure. The median of the mean age of blood units transfused per patient was 11 days [interquartile range, 8, 15] in the freshest available blood group and 23 days [interquartile range, 17, 30] in the oldest available blood group. In the primary analysis, posttransfusion acute kidney injury was observed in 688 of 4,777 (14.4%) patients given the freshest available blood and 1,487 of 9,684 (15.4%) patients given the oldest available blood, with an estimated relative risk (95% CI) of 0.94 (0.86 to 1.02; P = 0.132). The secondary analysis treated blood age as a continuous variable (defined as duration of storage in days), with an estimated relative risk (95% CI) of 1.00 (0.96 to 1.04; P = 0.978) for a 10-day increase in the mean age of erythrocyte units. Conclusions In a population of patients without severely impaired baseline renal function receiving fewer than 10 erythrocyte units, duration of blood storage had no effect on the incidence of posttransfusion acute kidney injury. Editor’s Perspective What We Already Know about This Topic What This Article Tells Us That Is New

Purpose: While there is concern about degenerative tissue effects of exposure to space radiation during deep-space missions, there are no pharmacological countermeasures against these adverse effects. γ-Tocotrienol (GT3) is a natural form of vitamin E that has anti-oxidant properties, modifies cholesterol metabolism, and has anti-inflammatory and endothelial cell protective properties. The purpose of this study was to test whether GT3 could mitigate cardiovascular effects of oxygen ion (16O) irradiation in a mouse model. Materials and methods: Male C57BL/6 J mice were exposed to whole-body 16O (600 MeV/n) irradiation (0.26–0.33 Gy/min) at doses of 0 or 0.25 Gy at 6 months of age and were followed up to 9 months after irradiation. Animals were administered GT3 (50 mg/kg/day s.c.) or vehicle, on Monday – Friday starting on day 3 after irradiation for a total of 16 administrations. Ultrasonography was used to measure in vivo cardiac function and blood flow parameters. Cardiac tissue remodeling and inflammatory infiltration were assessed with histology and immunoblot analysis at 2 weeks, 3 and 9 months after radiation. Results: GT3 mitigated the effects of 16O radiation on cardiac function, the expression of a collagen type III peptide, and markers of mast cells, T-cells and monocytes/macrophages in the left ventricle. Conclusions: GT3 may be a potential countermeasure against late degenerative tissue effects of high-linear energy transfer radiation in the heart.

BACKGROUND Tomatoes contain carotenoids that have the potential to alter the effects of external beam radiation therapy (EBRT). OBJECTIVES We hypothesized that dietary lyophilized tomato paste (TP) would reduce apoptosis within carotenoid-containing nonneoplastic tissues in EBRT-treated TRansgenic Adenocarcinoma of the Mouse Prostate (TRAMP) mice. METHODS Male TRAMP mice (n = 73) were provided an AIN-93G diet or a modified AIN-93G diet containing 10% TP (wt:wt) at 4 wk of age. Prostate tumor growth was monitored by ultrasound. The caudal half of the mouse was irradiated with 7.5 Gy (Rad) or 0 Gy (sham) at 24 wk of age or after the tumor volume exceeded 1000 mm3 with a Cobalt-60 source. Mice were euthanized 24 h postradiation. Carotenoids and α-tocopherol were measured by HPLC and compared by a t test. Tissues were assessed for radiation-induced changes (hematoxylin and eosin) and apoptosis [cleaved caspase-3 (CC3)] and compared by Kruskal-Wallis test or Freedman-Lane's permutation test. RESULTS Serum concentrations of lycopene (52% lower), phytoene (26% lower), and α-tocopherol (22% lower) were decreased in TP-fed irradiated mice (TP-Rad) compared with TP-fed sham mice (P

Powder technology has been used to coat fibers with polymeric particles to manufacture composites. The material parameters such as particle size and fiber diameter and process parameters such as the particle charge and the particle velocity will influence the powder coating process. In this work, we present an experimental setup in which we characterize the coating of micron size particles on a fiber and examine the role of process parameters on the coating density. Monodispersed Polystyrene (PS) particles (2.8 ± 0.4 μm in diameter) were used to coat a carbon fiber (CF) 7 μm in diameter. A model setup was designed and fabricated in which the PS particles were charged with an ozone charging system, while the CF was maintained at the grounded state at all times. The setup consisted of two parts namely, a cyclone and a coating chamber. The distance between the PS particles was measured and the density of coating was examined using a microscope as a function of the processing parameters used in the setup. The measured average distance between the PS particles on the CF was used in our developed numerical model to estimate the friction coefficient between the charged PS particle and the grounded CF and the fiber volume fraction. The proposed methodology can be extended to any particle size, particle charge, and fiber diameter to estimate the friction coefficient and the fiber volume fraction at the microscale level.

This study examined the possible buffering effect of acid-base glass-ionomer cements. Commercial capsulated materials were used, namely: Riva Self Cure, Ketac Molar, Equia Forte and Chemfil Rock. Cements were prepared by vibratory mixing then cylinders (4 mm diameter, 6 mm height) prepared from them using metal moulds. They were cured in the moulds at 37 °C for one hour, then placed in 8 ml deionised water at 22 °C and stored for 24 h or 4 weeks. Then they were crushed to powders with a pestle and mortar. Ten samples per powder (0.05 g) were placed in individual 10 ml volumes of deionised water. For two sets of ten (24 h or 4 weeks old), 1.0 ml of 0.01 M HCl was added, equilibrated and the pH measured by calibrated electrode. For another two sets of ten (24 h or 4 weeks old), 1.0 ml of 0.01 M NaOH was added, equilibrated, and the pH measured. The pH of a control set of solutions (no cement powder) was also measured. Data were tested using 1-way ANOVA and Tukey HSD test. The original acid solution pH was 3.2, which rose to 4.4–5.5 depending on brand and age of cement. The original alkali solution pH was 11.3, which fell to 6.7–8.3 depending on the details of the cement. Differences from initial pH had Tukey HSD p values of 0.001 in all cases. It was concluded that acid-base glass-ionomers can act as solid-state buffers, a finding attributed to the carboxylic acid/carboxylate conjugate pairs within them.

Protein quality is an important component of protein intake to support growth, development, and maintenance of essential body tissues and functions. Therefore, protein quality should be emphasized as a key characteristic during protein food selection within the larger context of healthy dietary patterns, especially when considering the wide variance of protein quality across animal- and plant-based foods. However, the USDA Dietary Guidelines for Americans (DGA) do not address specific protein quality recommendations within their protein foods ounce equivalents guidance or as a component of Healthy U.S. Style, Healthy Vegetarian, and Healthy Mediterranean Style dietary patterns. In addition, the protein foods ounce equivalents within the DGA are not established on any obvious metabolic equivalency characteristic [i.e., energy, protein, or essential amino acid (EAA) content], which creates misleading messaging of equivalent functional and metabolic benefit across protein foods. EAA content is a key characteristic of protein quality and can be a practical focal point for protein intake recommendations and achieving healthy dietary patterns. This review discusses the importance of protein quality, the state of messaging within DGA recommendations, and proposes EAA density (i.e., EAA content relative to total energy) as one practical approach to improve current dietary recommendations. Two recent publications that evaluated the DGA protein foods ounce equivalents based on metabolic effect and their application within DGA recommended dietary patterns are discussed.

The GuLF STUDY is investigating health outcomes associated with oil spill-related chemical exposures among workers involved in the spill response and clean-up following the Deepwater Horizon disaster. Due to the lack of dermal exposure measurements, we estimated dermal exposures using a deterministic model, which we customized from a previously published model. Workers provided information on the frequency of contact with oil, tar, chemical dispersants applied to the oil spill and sea water, as well as the use of protective equipment, by job/activity/task. Professional judgment by industrial hygienists served as a source of information for other model variables. The model estimated dermal exposures to total hydrocarbons (THC), benzene, ethylbenzene, toluene, xylene, n-hexane (BTEX-H), polycyclic aromatic hydrocarbons (PAHs), and dispersants in GuLF DREAM units (GDUs). Arithmetic means (AMs) of THC exposure estimates across study participants ranged from 0.9) for most of the substances in oil but were lower for some of the substances in tar. These data were linked to the study participants to allow investigation of adverse health effects that may be related to dermal exposures.

The neocortex is disproportionately expanded in human compared with mouse1,2, both in its total volume relative to subcortical structures and in the proportion occupied by supragranular layers composed of neurons that selectively make connections within the neocortex and with other telencephalic structures. Single-cell transcriptomic analyses of human and mouse neocortex show an increased diversity of glutamatergic neuron types in supragranular layers in human neocortex and pronounced gradients as a function of cortical depth3. Here, to probe the functional and anatomical correlates of this transcriptomic diversity, we developed a robust platform combining patch clamp recording, biocytin staining and single-cell RNA-sequencing (Patch-seq) to examine neurosurgically resected human tissues. We demonstrate a strong correspondence between morphological, physiological and transcriptomic phenotypes of five human glutamatergic supragranular neuron types. These were enriched in but not restricted to layers, with one type varying continuously in all phenotypes across layers 2 and 3. The deep portion of layer 3 contained highly distinctive cell types, two of which express a neurofilament protein that labels long-range projection neurons in primates that are selectively depleted in Alzheimer’s disease4,5. Together, these results demonstrate the explanatory power of transcriptomic cell-type classification, provide a structural underpinning for increased complexity of cortical function in humans, and implicate discrete transcriptomic neuron types as selectively vulnerable in disease., Combined patch clamp recording, biocytin staining and single-cell RNA-sequencing of human neurocortical neurons shows an expansion of glutamatergic neuron types relative to mouse that characterizes the greater complexity of the human neocortex.

Background: Canine cognitive dysfunction (CCD) is a progressive syndrome recognized in mature to aged dogs with a variety of neuropathological changes similar to human Alzheimer’s disease (AD), for which it is thought to be a good natural model. However, the presence of hyperphosphorylated tau protein (p-Tau) in dogs with CCD has only been demonstrated infrequently. Objective: The aim of the present study was to investigate the presence of p-Tau and amyloid-β oligomer (Aβo) in cerebral cortex and hippocampus of dogs with CCD, with focus on an epitope retrieval protocol to unmask p-Tau. Methods: Immunohistochemical and immunofluorescence analysis of the cortical and hippocampal regions of five CCD-affected and two nondemented aged dogs using 4G8 anti-Aβp, anti-Aβ1 - 42 nanobody (PrioAD13) and AT8 anti-p-Tau (Ser202, Thr205) antibody were used to demonstrate the presence of Aβ plaques (Aβp) and Aβ1 - 42 oligomers and p-Tau deposits, respectively. Results: The extracellular Aβ senile plaques were of the diffuse type which lack the dense core normally seen in human AD. While p-Tau deposits displayed a widespread pattern and closely resembled the typical human neuropathology, they did not co-localize with the Aβp. Of considerable interest, however, widespread intraneuronal deposition of Aβ1 - 42 oligomers were exhibited in the frontal cortex and hippocampal region that co-localized with p-Tau. Conclusion: Taken together, these findings reveal further shared neuropathologic features of AD and CCD, supporting the case that aged dogs afflicted with CCD offer a relevant model for investigating human AD.

In this study a numerical analysis of hydrogen production via an autothermal reforming reactor is presented. The endothermic reaction of steam methane reforming and the exothermic combustion of methane were activated with patterned Ni/Al2O3 catalytic layer and patterned Pt/Al2O3 catalytic layer, respectively. Aiming to achieve a more compacted process, a novel design of a reactor was proposed in which the reforming and the combustion catalysts were modeled as patterned thin layers. This configuration is analyzed and compared with two configurations. In the first configuration, the catalysts are modeled as continuous thin layers in parallel, while, in the second configuration the catalysts are modeled as continuous thin layers in series (conventional catalytic autothermal reactor). The results show that the pattern of the catalyst layers improves slightly the hydrogen yield, i.e. 3.6%. Furthermore, for the same concentration of hydrogen produced, the activated zone length can be decreased by 38% and 15% compared to the conventional catalytic autothermal reforming and the configuration where the catalysts are fitted in parallel, respectively. Besides, the oxygen consumption is lowered by 5%. The decrement of the catalyst amount and the oxygen feedstock in the novel studied design lead to lower costs and compact process. © 2021 Hydrogen Energy Publications LLC

This study describes a novel series of UDP-N-acetylglucosamine acyltransferase (LpxA) inhibitors that was identified through affinity-mediated selection from a DNA-encoded compound library. The original hit was a selective inhibitor of Pseudomonas aeruginosa LpxA with no activity against Escherichia coli LpxA. The biochemical potency of the series was optimized through an X-ray crystallography-supported medicinal chemistry program, resulting in compounds with nanomolar activity against P. aeruginosa LpxA (best half-maximal inhibitory concentration (IC50) 20 μM and MIC > 128 μg/mL). The mode of action of analogues was confirmed through genetic analyses. As expected, compounds were active against multidrug-resistant isolates. Further optimization of pharmacokinetics is needed before efficacy studies in mouse infection models can be attempted. To our knowledge, this is the first reported LpxA inhibitor series with selective activity against P. aeruginosa.

Biotransformation may substantially reduce the extent to which organic environmental contaminants accumulate in fish. Presently, however, relatively little is known regarding the biotransformation of ionized chemicals, including cationic surfactants, in aquatic organisms. To address this deficiency, a rainbow trout liver S9 substrate depletion assay (RT-S9) was used to measure in vitro intrinsic clearance rates (CLint ; ml min-1 g liver-1 ) for 22 cationic surfactants that differ with respect to alkyl chain length and degree of methylation on the charged nitrogen atom. None of the quaternary N,N,N-trimethylalkylammonium compounds exhibited significant clearance. Rapid clearance was observed for N,N-dimethylalkylamines, and slower rates of clearance were measured for N-methylalkylamine analogs. Clearance rates for primary alkylamines were generally close to or below detectable levels. For the N-methylalkylamines and N,N-dimethylalkylamines, the highest CLint values were measured for C10 -C12 homologs; substantially lower clearance rates were observed for homologs containing shorter or longer carbon chains. Based on its cofactor dependency, biotransformation of C12 -N,N-dimethylamine appears to involve one or more cytochrome P450-dependent reaction pathways, and sulfonation. On a molar basis, N-demethylation metabolites accounted for up to 25% of the N,N-dimethylalkylamines removed during the 2-h assay, and up to 55% of the removed N-methylalkylamines. These N-demethylation products possess greater metabolic stability in the RT-S9 assay than the parent structures from which they derive and may contribute to the overall risk of ionizable alkylamines. The results of these studies provide a set of consistently determined CLint values that may be extrapolated to whole trout to inform in silico bioaccumulation assessments. Environ Toxicol Chem 2021;40:3123-3136. © 2021 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Soft-ballistic sub-laminate (SBSL) made from ultra-high molecular weight polyethylene (UHMWPE) fibers in [0/90] stacking sequence are the building block of a multi-layer soft-ballistic armor pack (SBAP, aka Soft Armor). A systematic study of the perforation dynamics of a single layer SBSL and several multi-layer SBAPs (2, 3, 4, 8, 16, 24, 32 layers) is presented for the first time in the literature. A previously validated finite element model of transverse impact on a single layer is used to study the perforation mechanics of multi-layer SBAPs with friction between individual layers. Following the classical definition of ballistic limit velocity, a minimum perforation velocity has been determined for free-standing single layer SBSL and multi-layer SBAPs. For the multi-layer SBAPs, complete perforations have been identified as progressive perforation of individual layers through the thickness. The minimum perforation velocities of multi-layer SBAPS is linear with the areal density for the eight (8) layer target and thicker. Large deformation behavior and perforation mechanics of the SBAPs is discussed in detail.

Natural (RRR-) α-tocopherol (αT) is more bioactive than synthetic (all racemic, all rac-) αT, but not enough is known about the tissue kinetics of the 2 αT sources. We examined the time-course bioaccumulation of natural versus synthetic αT in tissues of young, marginally vitamin E-deficient mice using 13C-RRR-αT or 13C-all rac-αT tracers. In experiment 1, 3-week old male wild-type mice were fed a vitamin E-deficient diet for 0, 1, 2, or 3 weeks (n = 5/time point). Tissue αT levels were analyzed by HPLC-PDA. Feeding a vitamin E-deficient diet for up to 3 weeks decreased total αT concentrations in all analyzed tissues except the brain, which maintained its αT level. In experiment 2, a 2-week αT-depletion period was followed by administration of a single oral dose of 0.5 mg of 13C-RRR-αT or 13C-all rac-αT. At 12 hr, 1, 2, and 4 days post-dose, serum and multiple tissues were collected (n = 3/time point). αT was quantified by HPLC-PDA, and 13C-αT enrichment was determined by LC-MS. Both sources of 13C-αT reached maximum serum levels at 12 hr post-dose. 13C-RRR-αT levels were significantly higher than 13C-all rac-αT in serum at 1 d post-dose, and in heart, lungs, and kidney at 2d post-dose. In brain, 13C-RRR-αT concentrations were significantly higher than 13C-all rac-αT at 2 and 4 d post-dose. At 4 d post-dose, 13C-αT levels were similar between the 2 sources in examined tissues except for brain and adipose tissue where 13C-RRR-αT was higher. In conclusion, αT bioaccumulation over time varied substantially depending on αT source and tissue type.

Summary Communication of information through the global switching of conformation in synthetic molecules has hitherto entailed the inversion of chirality. Here, we report a class of oligomer through which information may be communicated through a global reversal of polarity. Ethylene-bridged oligoureas are constitutionally symmetrical, conformationally flexible molecules organized by a single chain of hydrogen bonds running the full length of the oligomer. NMR reveals that this hydrogen-bonded chain may undergo a coherent reversal of directionality. The directional uniformity of the hydrogen-bond chain allows it to act as a channel for the spatial communication of information on a molecular scale. A binding site at the terminus of an oligomer detects local information about changes in pH or anion concentration and transmits that information—in the form of a directionality switch in the hydrogen-bond chain—to a remote polarity-sensitive fluorophore. This propagation of polarity-encoded information provides a new mechanism for molecular communication., Graphical abstract, Highlights • Simple urea oligomers organize themselves with a single coherent hydrogen-bond chain • Reversing the polarity of the hydrogen bonding sends a message through the molecule • Conformational messages can be transmitted by changes in pH or by anion binding • Receipt of the conformational message is indicated by a remote fluorescent response, The bigger picture Despite the progress made by synthetic chemistry in building the molecular structures of nature, there are still domains of molecular science where biology is immensely more sophisticated than even the most advanced artificial chemical systems. One of these is the ability to use molecular structures to encode and communicate information. Nature stores information in the hydrogen-bond polarity of the base pairs and translates it into molecular function using the polarity matching of these bonds. Here, we outline an early step toward replicating this ability in the design, synthesis, and operation of a series of synthetic molecular devices that employ a structurally simple family of oligomeric molecules that can communicate information in the form of hydrogen-bond polarity. This study marks a significant step in the design of molecular systems that manipulate information, which will form the basis of the many compartmentalization-based nanotechnologies of the future., Information in nature is stored, communicated, and amplified by matching hydrogen-bond polarity in base pairs. Here, we show that structurally simple synthetic molecules can also use hydrogen-bond polarity to communicate information. Molecules with coherent chains of hydrogen bonds respond to changes in their environment by reversing the polarity of the chain, allowing messages to be communicated from one end of the molecule to the other. This work reveals a new mechanism for information handling in synthetic molecular systems.

Powder compaction-induced surface chemistry in metal oxide nanocrystal ensembles is important for very diverse fields such as triboelectrics, tribocatalysts, surface abrasion, and cold sintering of ceramics. Using a range of spectroscopic techniques, we show that MgO nanocube powder compaction with uniaxial pressures that can be achieved by gentle manual rubbing or pressing (p ≥ 5 MPa) excites energetic electron-hole pairs and generates oxygen radicals at interfacial defect structures. While the identification of paramagnetic O- radicals and their adsorption complexes with O2 point to the emergence of hole centers, triboemitted electrons become scavenged by molecular oxygen to convert into adsorbed superoxide anions O2 - as measured by electron paramagnetic resonance (EPR). By means of complementary UV-photoexcitation experiments, we found that photon energies in the range between 3 and 6 eV produce essentially the same EPR spectroscopic fingerprints and optical absorption features. To provide insights into this effect, we performed density functional theory calculations to explore the energetics of charge separation involving the ionization of low-coordinated anions and surface-adsorbed O2 - radicals at points of contact. For all selected configurations, charge transfer is not spontaneous but requires an additional driving force. We propose that a plausible mechanism for oxygen radical formation is the generation of significant surface potential differences at points of contact under loading as a result of the highly inhomogeneous elastic deformations coupled with the flexoelectric effect.

Funder: Gerald Kerkut Charitable Trust; FundRef: http://dx.doi.org/10.13039/100012166, Funder: Rank Prize, Funder: NIHR Clinical Lectureship, Pregnancy 25-hydroxyvitamin D [25(OH)D] concentrations are associated with maternal and fetal health outcomes. Using physiological human placental perfusion and villous explants, we investigate the role of the placenta in regulating the relationships between maternal 25(OH)D and fetal physiology. We demonstrate active placental uptake of 25(OH)D3 by endocytosis, placental metabolism of 25(OH)D3 into 24,25-dihydroxyvitamin D3 and active 1,25-dihydroxyvitamin D [1,25(OH)2D3], with subsequent release of these metabolites into both the maternal and fetal circulations. Active placental transport of 25(OH)D3 and synthesis of 1,25(OH)2D3 demonstrate that fetal supply is dependent on placental function rather than simply the availability of maternal 25(OH)D3. We demonstrate that 25(OH)D3 exposure induces rapid effects on the placental transcriptome and proteome. These map to multiple pathways central to placental function and thereby fetal development, independent of vitamin D transfer. Our data suggest that the underlying epigenetic landscape helps dictate the transcriptional response to vitamin D treatment. This is the first quantitative study demonstrating vitamin D transfer and metabolism by the human placenta, with widespread effects on the placenta itself. These data demonstrate a complex interplay between vitamin D and the placenta and will inform future interventions using vitamin D to support fetal development and maternal adaptations to pregnancy., CS was funded by a Gerald Kerkut Charitable Trust studentship and BA by Rank Prize and University of Southampton Vice Chancellor���s Studentships plus the MRC. KMG was supported by the UK Medical Research Council (MC_UU_12011/4), the National Institute for Health Research (NIHR Senior Investigator [NF-SI-0515-10042], NIHR Southampton 1000DaysPlus Global Nutrition Research Group [17/63/154], and NIHR Southampton Biomedical Research Centre [IS-BRC-1215-20004]), British Heart Foundation (RG/15/17/3174) and the US National Institute on Aging of the National Institutes of Health (Award No. U24AG047867). KSJ was supported by the National Institute for Health Research (NIHR) Cambridge Biomedical Research Centre (ISBRC-1215-20014). The NIHR Cambridge Biomedical Research Centre is a partnership between Cambridge University Hospitals NHS Foundation Trust and the University of Cambridge, funded by the NIHR. The views expressed are those of the authors and not necessarily those of the NHS, the NIHR, or the Department of Health and Social Care. Experimental work performed by KSJ and FH at MRC. EWL was supported by Dr Ann Prentice (UK Medical Research Council U105960371). The SWS has been supported by grants from Medical Research Council (MRC) (4050502589 [MRC LEU]), Dunhill Medical Trust, British Heart Foundation, Food Standards Agency, National Institute for Health Research (NIHR) Southampton Biomedical Research Centre, University of Southampton and University Hospital Southampton NHS Foundation Trust, NIHR Oxford Biomedical Research Centre, University of Oxford, and the European Union���s Seventh Framework Programme (FP7/2007-2013), project EarlyNutrition, under grant agreement 289346 and the European Union���s Horizon 2020 research and innovation program (LIFECYCLE, grant agreement no. 733206). EC has been supported by the Wellcome Trust (201268/Z/16/Z) and an NIHR Clinical Lectureship. Work leading to these results was supported by the BBSRC (HDHL-Biomarkers, BB/P028179/1), as part of the ALPHABET project, supported by an award made through the ERA-Net on Biomarkers for Nutrition and Health (ERA HDHL), Horizon 2020 grant agreement number 696295. The proteomic analyses (SDG and AM) were financially supported by the National Institutes of Health (R21AI122389) and the Beckman Institute at the California Institute of Technology. This project has received funding from the European Union���s Horizon 2020 research and innovation program under the Marie Sk��odowska-Curie grant agreement InvADeRS no. 841172 to JMF. The electron microscopy image in Figure 2 was produced with help of the Biomedical imaging unit, Faculty of Medicine, University of Southampton.

Background: Colchicine reduces risk of cardiovascular events in patients post–myocardial infarction and in patients with chronic coronary disease. It remains unclear whether this effect is related to the time of onset of treatment following an acute coronary syndrome (ACS). Objectives: This study investigates risk for major adverse cardiovascular events in relation to history and timing of prior ACS, to determine whether the benefits of colchicine are consistent independent of prior ACS status. Methods: The LoDoCo2 (Low-Dose Colchicine 2) trial randomly allocated patients with chronic coronary disease to colchicine 0.5 mg once daily or placebo. The rate of the composite of cardiovascular death, spontaneous myocardial infarction, ischemic stroke, or ischemia-driven coronary revascularization was compared between patients with no prior, recent (6-24 months), remote (2-7 years), or very remote (>7 years) ACS; interaction between ACS status and colchicine treatment effect was assessed. Results: In 5,522 randomized patients, risk of the primary endpoint was independent of prior ACS status. Colchicine consistently reduced the primary endpoint in patients with no prior ACS (incidence: 2.8 vs 3.4 events per 100 person-years; hazard ratio [HR]: 0.81; 95% confidence interval [CI]: 0.52-1.27), recent ACS (incidence: 2.4 vs 3.3 events per 100 person-years; HR: 0.75; 95% CI: 0.51-1.10), remote ACS (incidence: 1.8 vs 3.2 events per 100 person-years, HR: 0.55; 95% CI: 0.37-0.82), and very remote ACS (incidence: 3.0 vs 4.3 events per 100 person-years, HR: 0.70; 95% CI: 0.51-0.96) (P for interaction = 0.59). Conclusions: The benefits of colchicine are consistent irrespective of history and timing of prior ACS. (The LoDoCo2 Trial: Low Dose Colchicine for secondary prevention of cardiovascular disease [LoDoCo2] ACTRN12614000093684)

Carbon dioxide (CO2) and methane (CH4) are natural and anthropogenic products that play a central role in the global carbon cycle and regulating Earth's climate. Applications utilizing laser absorption spectroscopy, which continuously measure concentrations and stable isotope ratios of these greenhouse gases, are routinely employed to measure the source and magnitude of atmospheric inputs. We developed a discrete sample introduction module (DSIM) to enable measurements of methane and CO2 concentrations and δ13C values from limited volume (5-100 mL) gas samples when interfaced with a commercially available cavity ring-down spectroscopy (CRDS) analyzer. The analysis has a dynamic range that spans six orders of magnitude from 100% analyte to the lower limit of instrument detection (2 ppm). We demonstrate system performance for methane by comparing concentrations and δ13C results from the DSIM-CRDS system and traditional methods for a variety of sample types, including low concentration (nanomolar CH4) seawater and high concentration (>90% CH4) natural gas. The expansive concentration range of the field-portable DSIM-CRDS system can measure enhances analytical performance for investigating methane and CO2 dynamics and, potentially, other gases measured by laser absorption spectroscopy.

Brain α2-containing GABA(A) receptors play a critical role in the modulation of anxiety- and fear-like behavior. However, it is unknown whether these receptors also play a role in modulating resilience to chronic stress, and in which brain areas and cell types such an effect would be mediated. We evaluated the role of α2-containing GABA(A) receptors following chronic social defeat stress using male mice deficient in the α2 subunit globally or conditionally in dopamine D1- or D2-receptor-expressing neurons, e.g., within the nucleus accumbens (NAc). In addition, we examined the effect of the lack of the α2 subunit on intermediates of the glutathione synthesis pathway. We found that α2-containing GABA(A) receptors on D2-receptor-positive but not on D1-receptor-positive neurons promote resiliency to chronic social defeat stress, as reflected in social interaction tests. The pro-resiliency effects of α2-containing GABA(A) receptors on D2-receptor-positive neurons do not appear to be directly related to alterations in anxiety-like behavior, as reflected in the elevated plus-maze, light–dark box, and novel open field tests. Increases in indices of oxidative stress—reflected by increases in cystathionine levels and reductions in GSH/GSSG ratios—were found in the NAc and prefrontal cortex but not in the hippocampus of mice lacking α2-containing GABA(A) receptors. We conclude that α2-containing GABA(A) receptors within specific brain areas and cell populations promote stress resiliency independently of direct effects on anxiety-like behaviors. A potential mechanism contributing to this increased resiliency is the protection that α2-containing GABA(A) receptors provide against oxidative stress in NAc and the prefrontal cortex.

Carbon–carbon composites (CCCs) are a unique form of carbon fiber-reinforced materials that exhibit excellent thermomechanical properties under extreme environmental conditions. Due to the need for the retention of mechanical properties at temperatures exceeding 2000 °C, CCCs have been utilized in heat shields, rocket nozzles, aircraft brakes, and leading edge material in hypersonic vehicles. In order to expand the applicability of CCCs, the fabrication process must be modified such that there is a reduction in cost or processing time. It is hypothesized that maximizing the permeability of the composite, during processing, will grant the largest reduction in the fabrication time as it leads to a larger volume of pores filled during re-densification. This review attempts to capture the various parameters that have led to increased permeability, as well as outlining process modifications that have demonstrated influence over the carbonized microstructure. In addition, this review seeks to differentiate itself by systematically outlining research advances that have been made in each step of the fabrication process. In doing so, scientific gaps that exist can be expounded upon while simultaneously summarizing what is necessary to advance the field.

BACKGROUND: Blood clot contraction, volume shrinkage of the clot, is driven by platelet contraction and accompanied by compaction of the erythrocytes and their gradual shape change from biconcave to polyhedral, with the resulting cells named polyhedrocytes. OBJECTIVES: Here, we examined the role of erythrocyte rigidity on clot contraction and erythrocyte shape transformation. METHODS: We used an optical tracking methodology that allowed us to quantify changes in contracting clot size over time. RESULTS AND CONCLUSIONS: Erythrocyte rigidity has been shown to be increased in sickle cell disease (SCD), and in our experiments erythrocytes from SCD patients were 4-fold stiffer than those from healthy subjects. On average, the final extent of clot contraction was reduced by 53% in the clots from the blood of patients with SCD compared to healthy individuals, and there was significantly less polyhedrocyte formation. To test if this reduction in clot contraction was due to the increase in erythrocyte rigidity, we used stiffening of erythrocytes via chemical cross-linking (glutaraldehyde), rigidifying Wright(b) antibodies (Wr(b)), and naturally more rigid llama ovalocytes. Results revealed that stiffening erythrocytes result in impaired clot contraction and fewer polyhedrocytes. These results demonstrate the role of erythrocyte rigidity in the contraction of blood clots and suggest that the impaired clot contraction/shrinkage in SCD is due to the reduced erythrocyte deformability, which may be an underappreciated mechanism that aggravates obstructiveness of erythrocyte- rich (micro)thrombi in SCD.

Summary In order to better differentiate ante-mortem lesions from post-mortem retinal autolysis, the temporal sequence of post-mortem changes was studied in a well-controlled mouse model. Mice were of the same strain, age and sex, and were held at a constant ambient temperature. Eyes were collected at various times up to 72 h after death and immersion-fixed in either Davidson's fixative or 10% neutral buffered formalin, paraffin-embedded and sections cut and stained with haematoxylin and eosin. The most prominent, and early, autolytic change was retinal detachment, and subsequent folding, which occurred immediately after death in formalin-fixed eyes, but not until 2 h post mortem with Davidson's fixative. Retinal separation was complete at 16 h, or almost complete by 2 h, in formalin, but in Davidson's fixative, was only partial and segmental, the latter not becoming total until much later. Retinal detachment was attended by progressively more severe disruption and dissolution of photoreceptors and, particularly in Davidson's-fixed retinas, the rod outer segment often showed marked homogenization from 30 min to 4 h after death. The other major early change was nuclear pyknosis in the inner nuclear layer. Ganglion cells initially had cytoplasmic swelling, followed by shrinkage and basophilia (at 4 h with formalin and 16 h with Davidson's), with nuclear pyknosis becoming increasingly common over time. While the three retinal neuronal layers eventually became more attenuated and depleted of cells, the thickness of these layers was augmented by severe swelling. These findings show that the post-mortem interval at which histological interpretation of retinal changes becomes potentially compromised is dependent on the duration of this interval and the fixative used.

Heat shock proteins (HSPs) are important mediators of the normal cellular function and the cellular stress response. As such, HSPs are often utilized to measure the effects of stressors on organisms in vivo. However, multiple variables can influence their expression, including time or season, confounding results. To investigate the utility of HSPs in measuring effects of stressors in a top-trophic carnivore, we captured 20 American Alligators (Alligator mississippiensis (Daudin, 1802)), placed them in burlap sacks for 2 h and collected blood samples over four time points (baseline, 1 and 2 h after placement into burlap sacks, and 24 h after initial capture) to measure plasma corticosterone (the main crocodilian glucocorticoid) and levels of HSP60, HSP70, and HSP90. Time point significantly affected plasma corticosterone levels in Alligators (p < 0.0001), with levels significantly elevated at 1, 2, and 24 h, relative to baseline (all p < 0.05). However, capture stress did not affect HSP60, HSP70, or HSP90 in red blood cells (all p > 0.05). Our results suggest HSPs may be important biomarkers for investigating the impacts of stressors on captive and wild crocodilians, as they are not acutely elevated by capture or handling stress.

Gene therapy continues to be a promising contender for the treatment of cystic fibrosis (CF) airway disease. We have previously demonstrated that airway conditioning with lysophosphatidylcholine (L...

This study aimed to investigate the pharmacodynamic effects of indobufen and low-dose aspirin in patients with coronary atherosclerosis. In the first phase, 218 patients with coronary atherosclerosis were randomly assigned to receive aspirin 100 mg once daily (standard dose); 100 mg once every 2 days; 100 mg once every 3 days; 50 mg twice daily; 75 mg once daily; 50 mg once daily; or indobufen 100 mg twice daily for 1 month. In the second phase, 20 healthy subjects were treated with indobufen 100 mg twice daily for 1 week followed after a 2-week washout by aspirin 100 mg once daily for 1 week. The primary outcome was arachidonic acid-induced platelet aggregation (PLAA), and the secondary outcomes included plasma thromboxane B2 (TXB2) and urinary 11-dehydro-TXB2 (11-dh-TXB2) levels at the end of each treatment. In the first phase, compared with aspirin 100 mg once daily: all aspirin groups had similar suppression of PLAA whereas indobufen group had significantly less suppressed PLAA. Aspirin given every second or third day, and indobufen produced less suppression of plasma TXB2. All treatment regimens produced similar inhibition of 11-dh-TXB2. In the second phase, compared with aspirin, indobufen produced less suppression of plasma TXB2 at 8 h and 12 h after the last dose. Aspirin 50 mg twice daily, 75 mg once daily, and aspirin 50 mg once daily produce antiplatelet effects that are similar to aspirin 100 mg once daily. Aspirin given less often than once daily and indobufen 100 mg twice daily do not suppress platelets as effectively as aspirin 100 mg once daily.

ConspectusThe redox reaction pathway is crucial to the sustainable production of the fuels and chemicals required for a carbon-neutral society. Our society is becoming increasingly dependent on devices using batteries and electrolyzers, all of which rely on a series of redox reactions. The overall properties of oxide materials make them very well suited for such electrochemical and catalytic applications due to their associated cationic redox properties and the static site-adsorbate interactions. As these technologies have matured, it has become apparent that defect-driven redox reactions, defect-coupled diffusion, and structural transformations that are both time- and rate-dependent are also critical materials processes. This change in focus, considering not only redox properties but also more complex, dynamic behaviors, represents a new research frontier in the molecular sciences as they are strongly linked to device operation and degradation and lie at the heart of various phenomena that take place at electrochemical interfaces. Fundamental studies of the structural, electronic, and chemical transformation mechanisms are key to the advancement of materials and technological innovations that could be implemented in various electrochemical systems.In this Account, we focus on recent studies and advances in characterizing and understanding the dynamic redox evolution and structural transformations that take place in model perovskites and layered oxides under reactive conditions and correlate them with degradation mechanisms and operations in electrolyzers and batteries. We show that the dynamic evolution of oxygen vacancies and cationic migration in the surface or bulk occurs at the solid-liquid interface, using a combination of different synchrotron-based X-ray spectroscopies and scattering probes. Detailed redox-structure-reactivity correlation studies show how defects and diffusion processes can be tailored to drive various physical and chemical transformations in electrolyzers and batteries. We also highlight a strong correlation between oxygen redox reactivity and structural reorganization in both model thin films and particles, helping to bridge the gap between fundamental studies of the reaction mechanism and device applications. On the basis of these findings, we discuss strategies to probe and tune the redox reactivity and structural stability of the redox-active oxide interphase toward devising efficient pathways for energy and chemical harvesting.

Author(s): Wright, Erik; Hafner, John W.; Podolej, Gregory; Feinstein, Douglas L.; van Breemen, Richard; Rubinstein, Israel; Aks, Steven; Wahl, Michael | Abstract: Introduction: Synthetic cannabinoids are a rapidly expanding subset of designer drugs widely available in the United States since 2008. In Illinois during the spring of 2018, over 160 documented cases of bleeding and prolonged coagulopathy occurred secondary to contaminated synthetic cannabinoids. Methods: We conducted a retrospective cohort study consisting of 38 patients to describe the initial emergency department (ED) presentation, diagnosis, and treatment. Results: Through serum testing we found that three long-acting anticoagulant rodenticides (LAAR) were detected in patients who had inhaled these tainted products: brodifacoum, difenacoum, and bromodialone.n Discussion: This study encompasses the largest ED presentation of LAAR poisoning via the inhalational route known to date.nConclusion: The emergency physician should be aware of the potential for tainted coingestants as the cause of undifferentiated coagulopathy.

Isolated single wood fibres with cellulose fibril angles from 10 to 43° were tested in microtensile tests under controlled temperature and relative humidity of 5, 50, 75, 90% and in the wet state. This systematic study provides experimental stiffness and strength data, calculated on cell wall cross sections. It has been shown that stiffness reduction with increasing moisture content is more pronounced in fibres with large cellulose fibril angles. Interestingly, stiffness reduction in fibres with low cellulose fibril angles has been observed for the fully hydrated state only. The experimental dataset was fed into a model to determine moisture dependent stiffness of the hemicellulose-lignin-matrix and the stresses acting on the fibrils and the matrix.

Advances in multiplexed imaging technologies have drastically improved our ability to characterize healthy and diseased tissues at the single-cell level. Co-detection by indexing (CODEX) relies on DNA-conjugated antibodies and the cyclic addition and removal of complementary fluorescently labeled DNA probes and has been used so far to simultaneously visualize up to 60 markers in situ. CODEX enables a deep view into the single-cell spatial relationships in tissues and is intended to spur discovery in developmental biology, disease and therapeutic design. Herein, we provide optimized protocols for conjugating purified antibodies to DNA oligonucleotides, validating the conjugation by CODEX staining and executing the CODEX multicycle imaging procedure for both formalin-fixed, paraffin-embedded (FFPE) and fresh-frozen tissues. In addition, we describe basic image processing and data analysis procedures. We apply this approach to an FFPE human tonsil multicycle experiment. The hands-on experimental time for antibody conjugation is ~4.5 h, validation of DNA-conjugated antibodies with CODEX staining takes ~6.5 h and preparation for a CODEX multicycle experiment takes ~8 h. The multicycle imaging and data analysis time depends on the tissue size, number of markers in the panel and computational complexity. This protocol describes co-detection by indexing, a highly multiplexed imaging technology that uses DNA-conjugated antibodies to image up to 60 markers in formalin-fixed, paraffin-embedded and fresh-frozen tissues.

Objective The aim of the present study was to determine the chemical species formed inside glass-ionomer cements after fluoride uptake and to investigate the depth of penetration of fluoride ions within the cement matrix. Methods An experimental fluoride-free glass with composition 2SiO2–AlO3–CaO was produced. The glass powder was mixed with aqueous poly(acrylic acid) (PAA), and allowed to set. The resulting specimens were stored in 20 ml KF solution with 1000 ppm fluorine for 24 h and then placed into the same amount of water as for 24 h. A fluoride selective electrode was used to give the F concentration of the respective solutions. 19F MAS-NMR spectra were recorded on powdered cement specimens using a Bruker AVANCE-NEO 600 spectrometer. In addition, SEM observation and EDX chemical analysis were conducted on the cross-section of a carefully fractured specimen. Results Fluoride was shown to be mainly present in the surface layers of the specimen after placement in the KF solution, and only a small fraction was re-released into water. 19F NMR spectroscopy showed that AlF complexes were formed within the cement. Significance The fluoride taken up by a free-fluoride glass ionomer cement mostly occupies surface layers and is retained because it bonds to aluminum within the matrix. This finding explains why the majority of fluoride taken up by conventional glass ionomer cements is retained.

Human cytomegalovirus (HCMV) is currently a major cause of congenital disease in newborns and organ failure in transplant recipients. Despite decades of efforts, an effective vaccine against HCMV has yet to be developed. However, the discovery of pentameric gH complex on viral surface which contains potent neutralizing epitopes may help enable development of an effective vaccine. In our company ongoing Phase II clinical trial of whole-live virus HCMV vaccine (V160), the pentameric gH complex has been restored on the surface of live attenuated AD169 virus strain. The reconstructed HCMV virus contains a variety of surface glycoproteins including pentameric gH/gL/gUL128-131 complex, trimeric gH/gL/gO complex, gB glycoprotein, and gM/gN heterodimer complex. To further characterize this virus and enable the monitoring of multiple viral antigens during vaccine process development an effective and efficient analytical strategy was required to detect and quantify several viral surface proteins. In this paper, we present an innovative approach based on capillary western blot technology that allows fast and accurate quantitation of pentameric gH/gL/gUL128-131 complex, trimeric gH/gL/gO complex, and gB glycoprotein. This method is suitable for analyzing target proteins in multiple sample types including supernatants from infected cell culture, purification intermediates, concentration bulk, and the final vaccine product. In addition, the capillary western blot-based technology identified a previously unknown biochemical profile present in some HCMV viruses: triplet gH peaks of viral surface proteins in non-reducing environment, which could potentially present a new strategy for specificity and identity testing.

Objective Soluble fibrin (SF) is a substantial component of plasma fibrinogen (fg), but its composition, functions, and clinical relevance remain unclear. The study aimed to evaluate the molecular composition and procoagulant function(s) of SF. Materials and Methods Cryoprecipitable, SF-rich (FR) and cryosoluble, SF-depleted (FD) fg isolates were prepared and adsorbed on one hydrophilic and two hydrophobic surfaces and scanned by atomic force microscopy (AFM). Standard procedures were used for fibrin polymerization, crosslinking by factor XIII, electrophoresis, and platelet adhesion. Results Relative to FD fg, thrombin-induced polymerization of FR fg was accelerated and that induced by reptilase was markedly delayed, attributable to its decreased (fibrinopeptide A) FpA. FR fg adsorption to each surface yielded polymeric clusters and co-cryoprecipitable solitary monomers. Cluster components were crosslinked by factor XIII and comprised ≤21% of FR fg. In contrast to FD fg, FR fg adsorption on hydrophobic surfaces resulted in fiber generation enabled by both clusters and solitary monomers. This began with numerous short protofibrils, which following prolonged adsorption increased in number and length and culminated in surface-linked three-dimensional fiber networks that bound platelets. Conclusion The abundance of adsorbed protofibrils resulted from (1) protofibril/fg clusters whose fg was dissociated during adsorption, and (2) adsorbed des-AA monomers that attracted solution counterparts initiating protofibril assembly and elongation by their continued incorporation. The substantial presence of both components in transfused plasma and cryoprecipitate augments hemostasis by accelerating thrombin-induced fibrin polymerization and by tightly anchoring the resulting clot to the underlying wound or to other abnormal vascular surfaces.

The importance of real-time, quantitative toxicology data available for physicians treating poisoned patients was illustrated during the 2018 outbreak in Illinois of severe coagulopathy caused by inhaling illicit synthetic cannabinoids products contaminated with commercially-available brodifacoum, difenacoum, and bromadiolone, three potent, long-acting anticoagulant rodenticides (LAARs). Identification and quantification of these life-threatening toxins in blood samples of hospitalized patients required toxicology testing with liquid chromatography-tandem mass spectrometry (LC-MS/MS) that was not available in clinical laboratories of hospitals at the time of the outbreak. This highly-sensitive, quantitative assay can provide critical information to guide patient care during and after hospitalization, including identification of offending LAARs, estimates of the ingested dose, and dosage and discontinuation of oral vitamin K1 therapy after hospital discharge once plasma LAARs concentrations decreased to a safe level (