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Stellar streams are sensitive probes of the Galactic potential. The likelihood of a stream model given stream data is often assessed using simulations. However, comparing to simulations is challenging when even the stream paths can be hard to quantify. Here we present a novel application of Self-Organizing Maps and first-order Kalman Filters to reconstruct a stream's path, propagating measurement errors and data sparsity into the stream path uncertainty. The technique is Galactic-model independent, non-parametric, and works on phase-wrapped streams. With this technique, we can uniformly analyze and compare data with simulations, enabling both comparison of simulation techniques and ensemble analysis with stream tracks of many stellar streams. Our method is implemented in the public Python package TrackStream, available at https://github.com/nstarman/trackstream., 16 pages, 11 figures, preprint

The cell-penetrating peptide NAF-1 has recently emerged as a promising candidate for selective penetration and destruction of cancer cells. It displays numerous membrane-selective behaviors including cell-specific uptake and organelle-specific degradation. In this work, we explore membrane penetration and translocation of NAF-1 in model lipid bilayer vesicles as a function of lipid identity in zwitterionic phosphatidylcholine lipids mixed with anionic phosphatidylserine, phosphatidylglycerol, and phosphatidic acid lipids. By monitoring the digestion of NAF-1 using the protease trypsin located inside but not outside the vesicles, we determined that the translocation of NAF-1 was significantly enhanced by the presence of phosphatidic acid in the membrane compared to the other three anionic or zwitterionic lipids. These findings were correlated to fluorescence measurements of dansyl-labeled NAF-1, which revealed whether noncovalent interactions between NAF-1 and the bilayer were most stable either at the membrane/solution interface or within the membrane interior. Phosphatidic acid promoted interactions with fatty acid tails, while phosphatidylcholine, phosphatidylserine, and phosphatidylglycerol stabilized interactions with polar lipid headgroups. Interfacial vibrational sum frequency spectroscopy experiments revealed that the phosphate moiety on phosphatidic acid headgroups was better hydrated than on the other three lipids, which helped to shuttle NAF-1 into the hydrophobic region. Our findings demonstrate that permeation does not depend on the net charge on phospholipid lipid headgroups in these model vesicles and suggest a model wherein NAF-1 crosses membranes selectively due to lipid-specific interactions at bilayer surfaces.

By the late 1980s, widespread dramatic declines in amphibian populations were causing alarm. The culprit was identified as Batrachochytrium dendrobatidis (Bd), a chytrid fungus that infects the skin of various amphibian hosts, particularly anurans (frogs), and the first example of a chytridiomycete parasitising vertebrates. The disease, chytridiomycosis, has spread globally and is linked to the decline and extinction of many amphibian species. This review summarises the discovery of Bd, its emergence as a panzootic pathogen, and some current mitigation strategies to conserve amphibians.

Immobilizing enzymes onto abiological surfaces is a key step for developing protein-based technologies that can be useful for applications such as biosensors and biofuel cells. A central impediment for the advancement of this effort is a lack of generalizable strategies for functionalizing surfaces with proteins in ways that prevent unfolding, aggregation, and uncontrolled binding, requiring surface chemistries to be developed for each surface-enzyme pair of interest. In this work, we demonstrate a significant advancement toward addressing this problem using a gold nanoparticle (AuNP) as an initial scaffold for the chemical bonding of the enzyme acetylcholinesterase (AChE), forming the conjugate AuNP-AChE. This can then be placed onto chemically and structurally distinct surfaces (e.g., metals, semiconductors, plastics

Preventing SARS-CoV-2 infection by modulating viral host receptors, such as angiotensin-converting enzyme 2 (ACE2)1, could represent a new chemoprophylactic approach for COVID-19 that complements vaccination2,3. However, the mechanisms that control the expression of ACE2 remain unclear. Here we show that the farnesoid X receptor (FXR) is a direct regulator of ACE2 transcription in several tissues affected by COVID-19, including the gastrointestinal and respiratory systems. We then use the over-the-counter compound z-guggulsterone and the off-patent drug ursodeoxycholic acid (UDCA) to reduce FXR signalling and downregulate ACE2 in human lung, cholangiocyte and intestinal organoids and in the corresponding tissues in mice and hamsters. We show that the UDCA-mediated downregulation of ACE2 reduces susceptibility to SARS-CoV-2 infection in vitro, in vivo and in human lungs and livers perfused ex situ. Furthermore, we reveal that UDCA reduces the expression of ACE2 in the nasal epithelium in humans. Finally, we identify a correlation between UDCA treatment and positive clinical outcomes after SARS-CoV-2 infection using retrospective registry data, and confirm these findings in an independent validation cohort of recipients of liver transplants. In conclusion, we show that FXR has a role in controlling ACE2 expression and provide evidence that modulation of this pathway could be beneficial for reducing SARS-CoV-2 infection, paving the way for future clinical trials.

Three-body interactions can eject stars from the core of a globular cluster, causing them to enter the Galactic halo as extra-tidal stars. While finding extra-tidal stars is imperative for understanding cluster evolution, connecting isolated extra-tidal field stars back to their birth cluster is extremely difficult. In this work, we present a new methodology consisting of high-dimensional data analysis and a particle spray code to identify extra-tidal stars of any Galactic globular cluster using M3 as a case study. Using the t-Stochastic Neighbour Embedding (t-SNE) and Uniform Manifold Approximation and Projection (UMAP) machine learning dimensionality reduction algorithms, we first identify a set of 103 extra-tidal candidates in the APOGEE DR17 data catalogue with chemical abundances similar to M3 stars. To confirm each candidate's extra-tidal nature, we introduce corespray - a new Python-based three-body particle spray code that simulates extra-tidal stars for any Galactic globular cluster. Using Gaia EDR3 proper motions and APOGEE DR17 radial velocities, we apply multivariate Gaussian modelling and an extreme deconvolution to identify the extra-tidal candidates that are more likely to be associated with a distribution of corespray-simulated M3 extra-tidal stars than the field. Through these methods, we identify 10 new high-probability extra-tidal stars produced via three-body interactions in M3. We also explore whether any of our extra-tidal candidates are consistent with being ejected from M3 through different dynamical processes. Future applications of corespray will yield better understandings of core dynamics, star formation histories and binary fractions in globular clusters., Comment: 16 pages, 8 figures, 3 tables. Accepted for publication in Monthly Notices of the Royal Astronomical Society

Cardiovascular disease is a complication of systemic inflammatory diseases and anti-neutrophil cytoplasm antibody-associated vasculitis (AAV) is an exemplar. Mechanisms are poorly understood, and risk-reduction strategies are lacking.In a series of double-blind, randomized case-control forearm plethysmography and crossover systemic interventional studies, we examined arterial stiffness and endothelial function in AAV patients in long-term disease remission and in matched healthy volunteers (n=32/group). The primary outcome for the case-control study was the difference in endothelium-dependent vasodilation between health and AAV, and for the crossover study was the difference in pulse wave velocity (PWV) between treatment with placebo and selective endothelin-A receptor antagonism. Parallel in vitro studies of circulating monocytes and platelets explored mechanism.Case-control study: Compared to healthy volunteers, AAV patients had ~30% reduced endothelium-dependent vasodilation (mean difference: -6.1 mL/100mL of tissue/min, pCrossover study: Compared to placebo, both selective endothelin-A and dual endothelin-A/B receptor blockade reduced PWV and increased tPA release in AAV. Mechanistically, patients with AAV had increased platelet activation, more platelet-monocyte aggregates, and altered monocyte endothelin receptor function, reflecting reduced endothelin-1 clearance. AAV patients in long-term remission have elevated cardiovascular risk. Endothelin-1 contributes, and our data support a role for endothelin-blockers to reduce this risk by reducing arterial stiffness and increasing circulating tPA activity.

A physical understanding of membrane permeation and translocation by small, positively charged molecules can illuminate cell penetrating peptide mechanisms of entry and inform drug design. We have previously investigated the permeation of the doubly charged peptide WKW and proposed a defect-assisted permeation mechanism where a small molecule with +2 charge can achieve a metastable state spanning the bilayer by forming a membrane defect with charges stabilized by phospholipid phosphate groups. Here, we investigate the membrane permeation of two doubly charged peptides, WWK and WWWK, with charges separated by different lengths. Through complementary experiments and molecular dynamics simulations, we show that membrane permeation was an order of magnitude more favorable when charges were separated by an ∼2-3 Å greater distance on WWWK compared to WWK. These results agree with the previously proposed defect-assisted permeation mechanism, where a greater distance between positive charges would require a less extreme membrane defect to stabilize the membrane-spanning metastable state. We discuss the implications of these results in understanding the membrane permeation of cell-penetrating peptides and other small, positively charged membrane permeants.

PurposeAMB-FUBINACA is a synthetic cannabinoid receptor agonist (SCRA) which is primarily metabolised by hepatic enzymes producing AMB-FUBINACA carboxylic acid. The metabolising enzymes associated with this biotransformation remain unknown. This study aimed to determine if AMB-FUBINACA metabolism could be reduced in the presence of carboxylesterase (CES) inhibitors and recreational drugs commonly consumed with it. The affinity and activity of the AMB-FUBINACA acid metabolite at the cannabinoid type-1 receptor (CB1) was investigated to determine the activity of the metabolite.MethodsThe effect of CES1 and CES2 inhibitors, and delta-9-tetrahydrocannabinol (Δ9-THC) on AMB-FUBINACA metabolism were determined using both human liver microsomes (HLM) and recombinant carboxylesterases. Radioligand binding and cAMP assays comparing AMB-FUBINACA and AMB-FUBINACA acid were carried out in HEK293 cells expressing human CB1.ResultsAMB-FUBINACA was rapidly metabolised by HLM in the presence and absence of NADPH. Additionally, CES1 and CES2 inhibitors both significantly reduced AMB-FUBINACA metabolism. Furthermore, digitonin (100 µM) significantly inhibited CES1-mediated metabolism of AMB-FUBINACA by ~ 56%, while the effects elicited by Δ9-THC were not statistically significant. AMB-FUBINACA acid produced only 26% radioligand displacement consistent with low affinity binding. In cAMP assays, the potency of AMB-FUBINACA was ~ 3000-fold greater at CB1as compared to the acid metabolite.ConclusionsCES1A1 was identified as the main hepatic enzyme responsible for the metabolism of AMB-FUBINACA to its less potent carboxylic acid metabolite. This biotransformation was significantly inhibited by digitonin. Since other xenobiotics may also inhibit similar SCRA metabolic pathways, understanding these interactions may elucidate why some users experience high levels of harm following SCRA use.

We recently derived, using the density-of-states approximation, analytic distribution functions for the outcomes of direct single-binary scatterings. Using these outcome distribution functions, we present in this paper a self-consistent statistical mechanics-based analytic model obtained using the Fokker–Planck limit of the Boltzmann equation. Our model quantifies the dominant gravitational physics, combining both strong and weak single–binary interactions, which drives the time evolution of binary orbital parameter distributions in dense stellar environments. We focus in particular the distributions of binary orbital energies and eccentricities. We find a novel steady-state distribution of binary eccentricities, featuring strong depletions of both the highest and the lowest eccentricity binaries. In energy space, we compare the predictions of our analytic model to the results of numerical N-body simulations, and find that the agreement is good for the initial conditions considered here. This work is a first step towards the development of a fully self-consistent semi-analytic model for dynamically evolving binary star populations in dense stellar environments due to direct few-body interactions.

Historical in situ ocean temperature profile measurements are important for a wide range of ocean and climate research activities. A large proportion of the profile observations have been recorded using expendable bathythermographs (XBTs), and required bias corrections for use in climate change studies. It is generally accepted that the bias, and therefore bias correction, depends on the type of XBT used. However, poor historical metadata collection practices mean the XBT probe type information is often missing, for 59% of profiles between 1967 and 2000, limiting the development of reliable bias corrections. We develop a process of estimating missing instrument type metadata (the combination of both model and manufacturer) systematically, constructing a machine learning pipeline based on thorough data exploration to inform these choices. The predicted instrument type, where missing, will facilitate improved XBT bias corrections. The new approach improves the accuracy of the XBT type classification compared to previous approaches from a recall value of 0.75–0.94. We also develop an approach to account for the uncertainty associated with metadata assignments using ensembles of decision trees, which could feed into an ensemble approach to creating ocean temperature datasets. We describe the challenges arising from the nature of the dataset in applying standard machine learning techniques to the problem. We have implemented this in a portable, reproducible way using standard data science tools, with a view to these techniques being applied to other similar problems in climate science.

Current Brain Injury Guidelines (BIG) characterize patients with intracranial hemorrhage taking antiplatelet or anticoagulant agents as BIG 3 (the most severe category) regardless of trauma severity. This study assessed the risk of in-hospital mortality or need for neurosurgery in patients taking low-dose aspirin who otherwise would be classified as BIG 1.This was a retrospective study at an academic level 1 trauma center. Patients were included if they were admitted with traumatic intracerebral hemorrhage (tICH) and were evaluated by the BIG criteria. Exclusion criteria included indeterminate BIG status or patients with missing primary outcomes documentation. Patients were categorized as BIG 1, BIG 2, BIG 3, or BIG 1 on aspirin (patients with BIG 1 features taking low-dose aspirin). The primary endpoint was a composite of neurosurgical intervention and all-cause in-hospital mortality. Key secondary endpoints include rate of ICH progression, and ICU and hospital free days.A total of 1,520 patients met inclusion criteria. Median initial GCS was 14 (IQR 12, 15), injury severity scale score was 17 (IQR 10, 25) and abbreviated injury scale subscore head and neck (AISHead) was 3 (IQR 3, 4). The rate of the primary outcome for BIG 1, BIG 1 on aspirin, BIG 2, and BIG 3 was 1%, 2.2%, 1%, and 27%, respectively; the difference between BIG 1 on aspirin and BIG 3 was significant (p0.001).Patients taking low-dose aspirin with otherwise BIG 1-grade injuries experienced mortality and required neurosurgery significantly less often than other patients categorized as BIG 3. Inclusion of low-dose aspirin in the BIG criteria should be re-evaluated.Class III.

When designing and interpreting results from clinical trials evaluating treatments for children on the autism spectrum, a complicating factor is that most children receive a range of concurrent treatments. Thus, it is important to better understand the types and hours of interventions that participants typically receive as part of standard of care, as well as to understand the child, family, and geographic factors that are associated with different patterns of service utilization. In this multi-site study, we interviewed 280 caregivers of 6-to-11-year-old school-aged children on the autism spectrum about the types and amounts of interventions their children received in the prior 6 weeks. Reported interventions were coded as “evidence-based practice” or “other interventions,” reflecting the level of empirical support. Results indicated that children received a variety of interventions with varying levels of empirical evidence and a wide range of hours (0 to 79.3 hours/week). Children with higher autism symptom levels, living in particular states, and who identified as non-Hispanic received more evidence-based intervention hours. Higher parental education level related to more hours of other interventions. Children who were younger, had lower cognitive ability, and with higher autism symptom levels received a greater variety of interventions overall. Thus, based on our findings, it would seem prudent when designing clinical trials to take into consideration a variety of factors including autism symptom levels, age, cognitive ability, ethnicity, parent education and geographic location. Future research should continue to investigate the ethnic, racial, and socioeconomic influences on school-aged intervention services.

Molecular biology achieves control over complex reaction networks by means of molecular systems that translate a chemical input (such as ligand binding) into an orthogonal chemical output (such as acylation or phosphorylation). We present an artificial molecular translation device that converts a chemical input – the presence of chloride ions – into an unrelated chemical output: modulation of the reactivity of an imidazole moiety, both as a Brønsted base and as a nucleophile. The modulation of reactivity operates through the allosteric remote control of imidazole tautomer states. The reversible coordination of chloride to a urea binding site triggers a cascade of conformational changes in a chain of ethylene-bridged hydrogen-bonded ureas, switching the chain’s global polarity, that in turn modulates the tautomeric equilibrium of a distal imidazole, and hence its reactivity. Switching reactivities of active sites by dynamically controlling their tautomer states is an untapped strategy for building functional molecular devices with allosteric enzyme-like properties.

Accurate understanding of x-ray diagnostics is crucial for both interpreting high-energy-density experiments and testing simulations through quantitative comparisons. X-ray diagnostic models are complex. Past treatments of individual x-ray diagnostics on a case-by-case basis have hindered universal diagnostic understanding. Here, we derive a general formula for modeling the absolute response of non-focusing x-ray diagnostics, such as x-ray imagers, one-dimensional space-resolved spectrometers, and x-ray power diagnostics. The present model is useful for both data modeling and data processing. It naturally accounts for the x-ray crystal broadening. The new model verifies that standard approaches for a crystal response can be good approximations, but they can underestimate the total reflectivity and overestimate spectral resolving power by more than a factor of 2 in some cases near reflectivity edge features. We also find that a frequently used, simplified-crystal-response approximation for processing spectral data can introduce an absolute error of more than an order of magnitude and the relative spectral radiance error of a factor of 3. The present model is derived with straightforward geometric arguments. It is more general and is recommended for developing a unified picture and providing consistent treatment over multiple x-ray diagnostics. Such consistency is crucial for reliable multi-objective data analyses.

Extracellular vesicles (EVs) are lipid-membrane nanoparticles that are shed or secreted by many different cell types. The extracellular vesicle (EV) research community has rapidly expanded in recent years and are leading efforts to deepen our understanding of EV biological functions in human physiology and pathology. These insights are also providing a foundation on which future EV-based diagnostics and therapeutics are poised to positively impact human health. However, current limitations in our understanding of EV heterogeneity, cargo loading mechanisms and the nascent development of EV metrology are all areas that have been identified as important scientific challenges. The field of synthetic biology is also contending with the challenge of understanding biological complexity as it seeks to combine multidisciplinary scientific knowledge with engineering principles, to build useful and robust biotechnologies in a responsible manner. Within this context, cell-free systems have emerged as a powerful suite of in vitro biotechnologies that can be employed to interrogate fundamental biological mechanisms, including the study of aspects of EV biogenesis, or to act as a platform technology for medical biosensors and therapeutic biomanufacturing. Cell-free gene expression (CFE) systems also enable in vitro protein production, including membrane proteins, and could conceivably be exploited to rationally engineer, or manufacture, EVs loaded with bespoke molecular cargoes for use in foundational or translational EV research. Our pilot data herein, also demonstrates the feasibility of cell-free EV engineering. In this perspective we discuss the opportunities and challenges for accelerating EV research and healthcare applications with cell-free synthetic biology.

Chronic complications are a significant concern for people living with HIV/AIDS (PLWHA) infection. HIV-associated neurocognitive disorders (HAND) are prevalent in PLWHA. Yet, the efficacy of medications that penetrate the central nervous system (CNS) at preventing or slowing the progression of HAND remains largely unknown. The objective of this study was to determine whether high CNS penetration effectiveness (CPE) regimens improve neurocognitive test scores in PLWHA on combined antiretroviral therapy (cART). Primary literature evaluating cognitive outcomes based on CPE score of cART regimens in PLWHA was assembled from PubMed/Medline and EMBASE. Both randomized controlled trials and observational studies with at least 12 weeks of follow-up were included. A meta-analysis was conducted to calculate the standardized mean difference. Eight trials including a total of 3,303 patients with 13,103 person-years of follow-up were included in the systematic review. Four trials (

Barotropic (i.e., depth-uniform) coastal oceanic Kelvin waves can provide rapid teleconnections from climate and weather events in one location to remote regions of the globe. Studies suggest that barotropic Kelvin waves observed around Antarctica may provide a mechanism for rapidly propagating circulation anomalies around the continent, with implications for continental shelf temperatures along the West Antarctic Peninsula and thus Antarctic ice mass loss rates. However, how the propagation of Kelvin waves around Antarctica is influenced by features such as coastal geometry and variations in bathymetry remains poorly understood. Here we study the propagation of barotropic Antarctic Kelvin waves using a range of idealized model simulations. Using a single-layer linear shallow water model with 1∘ horizontal resolution, we gradually add complexity of continental configuration, realistic bathymetry, variable planetary rotation, and forcing scenarios, to isolate sources and sinks of wave energy and the mechanisms responsible. We find that approximately 75% of sub-inertial barotropic Kelvin wave energy is scattered away from Antarctica as other waves in one circumnavigation of the continent, due mostly to interactions with bathymetry. Super-inertial barotropic Kelvin waves lose nearly 95% of their energy in one circumpolar loop, due to interactions with both coastal geometry and bathymetry. These results help to explain why only sustained signals of low-frequency resonant barotropic Kelvin waves have been observed around Antarctica, and contribute to our understanding of the role of rapid, oceanic teleconnections in climate.

Overweight and obesity increase the risk of development and progression of knee osteoarthritis (OA), with higher levels of fat mass and lower levels of lean mass associated with poorer functional status. The aim was to assess changes in weight, body composition and physical function following weight loss or weight maintenance interventions in knee OA. A comprehensive search of four databases was conducted. The risk of bias was assessed using the Quality Criteria Checklist for primary research. Primary outcomes included weight, body composition and physical function; secondary outcomes were lipids, inflammatory biomarkers and muscle strength. Eleven studies were included utilizing

Insulin receptor (IR) signaling controls multiple facets of animal physiology. Maximally four insulins bind to IR at two distinct sites, termed site-1 and site-2. However, the precise functional roles of each binding event during IR activation remain unresolved. Here, we showed that IR incompletely saturated with insulin predominantly forms an asymmetric conformation and exhibits partial activation. The one insulin bound IR adopts a Γ-shaped conformation. The two insulins bound IR assumes a Ƭ-shaped conformation. One insulin is bound at site-1 and another insulin simultaneously contacts both sites-1 and −2 in the Ƭ-shaped IR dimer. We further demonstrated that the concurrent binding of four insulins to sites-1 and −2 prevents the formation of asymmetric IR and promotes the T-shaped symmetric, fully active state. Collectively, our results demonstrate how the synergistic binding of multiple insulins promotes optimal IR activation.

Enzyme activity is the basis for many biosensors where a catalytic event is used to detect the presence and amount of a biomolecule of interest. To create a practical point-of-care biosensor, these enzymes need to be removed from their native cellular environments and immobilized on an abiological surface to rapidly transduce a biochemical signal into an interpretable readout. This immobilization often leads to loss of activity due to unfolded, aggregated, or improperly oriented enzymes when compared to the native state. In this work, we characterize the formation and surface packing density of a stable monolayer of acetylcholinesterase (AChE) immobilized on a planar gold surface and quantify the extent of activity loss following immobilization. Using spectroscopic ellipsometry, we determined that the surface concentration of AChE on a saturated Au surface in a buffered solution was 2.77 ± 0.21 pmol cm

Contrast-induced nephropathy (CIN) is a major complication of imaging in patients with chronic kidney disease (CKD). The publication of an academic randomized controlled trial (RCT; n = 83) reporting oral (N)-acetylcysteine (NAC) to reduce CIN led to 70 clinical trials, 23 systematic reviews, and 2 large RCTs showing no benefit. However, no mechanistic studies were conducted to determine how NAC might work; proposed mechanisms included renal artery vasodilatation and antioxidant boosting. We evaluated the proposed mechanisms of NAC action in participants with healthy and diseased kidneys. Four substudies were performed. Two randomized, double-blind, placebo-controlled, three-period crossover studies (n = 8) assessed the effect of oral and intravenous (i.v.) NAC in healthy kidneys in the presence/absence of iso-osmolar contrast (iodixanol). A third crossover study in patients with CKD stage III (CKD3) (n = 8) assessed the effect of oral and i.v. NAC without contrast. A three-arm randomized, double-blind, placebo-controlled parallel-group study, recruiting patients with CKD3 (n = 66) undergoing coronary angiography, assessed the effect of oral and i.v. NAC in the presence of contrast. We recorded systemic (blood pressure and heart rate) and renal (renal blood flow (RBF) and glomerular filtration rate (GFR)) hemodynamics, and antioxidant status, plus biomarkers of renal injury in patients with CKD3 undergoing angiography. Primary outcome for all studies was RBF over 8 hours after the start of i.v. NAC/placebo. NAC at doses used in previous trials of renal prophylaxis was essentially undetectable in plasma after oral administration. In healthy volunteers, i.v. NAC, but not oral NAC, increased blood pressure (mean area under the curve (AUC) mean arterial pressure (MAP): mean difference 29 h⋅mmHg, P = 0.019 vs. placebo), heart rate (28 h⋅bpm, P 0.001), and RBF (714 h⋅mL/min, 8.0% increase, P = 0.006). Renal vasodilatation also occurred in the presence of contrast (RBF 917 h⋅mL/min, 12% increase, P = 0.005). In patients with CKD3 without contrast, only a rise in heart rate (34 h⋅bpm, P = 0.010) and RBF (288 h⋅mL/min, 6.0% increase, P = 0.001) occurred with i.v. NAC, with no significant effect on blood pressure (MAP rise 26 h⋅mmHg, P = 0.156). Oral NAC showed no effect. In patients with CKD3 receiving contrast, i.v. NAC increased blood pressure (MAP rise 52 h⋅mmHg, P = 0.008) but had no effect on RBF (151 h⋅mL/min, 3.0% increase, P = 0.470), GFR (29 h⋅mL/min/1.73m², P = 0.122), or markers of renal injury. Neither i.v. nor oral NAC affected plasma antioxidant status. We found oral NAC to be poorly absorbed and have no reno-protective effects. Intravenous, not oral, NAC caused renal artery vasodilatation in healthy volunteers but offered no protection to patients with CKD3 at risk of CIN. These findings emphasize the importance of mechanistic clinical studies before progressing to RCTs for novel interventions. Thousands were recruited to academic clinical trials without the necessary mechanistic studies being performed to confirm the approach had any chance of working.

Purpose Numerous clinical scoring tools exist for a variety of patient populations and disease states, but few tools provide information specifically designed for use by critical care pharmacists. The medication regimen complexity–intensive care unit (MRC-ICU) score was designed to provide high-level information about the complexity of critically ill patients’ medication regimens for use by critical care pharmacists. To date, implementation of this score in the electronic medical record (EMR) has not been reported. Summary Using an agile project management framework, the MRC-ICU score was rapidly implemented into an academic medical center’s EMR. The score is automatically calculated for all critically ill patients and is available for critical care pharmacists to triage patient review in their individual workflow. Reporting capabilities of the score also allow for granular complexity trending over time and between units, supplementing other objective measures of pharmacist workload. Conclusion The MRC-ICU score can be quickly implemented into the EMR for pharmacist use in real time. Future investigations into how pharmacists utilize this information and how to harness reporting capabilities for pharmacist workload assessment are warranted.

The urotensin-II (U-II) receptor (UT, nomenclature as agreed by the NC-IUPHAR Subcommittee on the Urotensin receptor [26, 36, 94]) is activated by the endogenous dodecapeptide urotensin-II, originally isolated from the urophysis, the endocrine organ of the caudal neurosecretory system of teleost fish [7, 93]. Several structural forms of U-II exist in fish and amphibians [94]. The goby orthologue was used to identify U-II as the cognate ligand for the predicted receptor encoded by the rat gene gpr14 [2, 20, 63, 69, 72]. Human urotensin-II, an 11-amino-acid peptide [20], retains the cyclohexapeptide sequence of goby U-II that is thought to be important in ligand binding [61, 53, 10]. This sequence is also conserved in the deduced amino-acid sequence of rat urotensin-II (14 amino-acids) and mouse urotensin-II (14 amino-acids), although the N-terminal is more divergent from the human sequence [19]. A second endogenous ligand for the UT has been discovered in rat [86]. This is the urotensin II-related peptide, an octapeptide that is derived from a different gene, but shares the C-terminal sequence (CFWKYCV) common to U-II from other species. Identical sequences to rat urotensin II-related peptide are predicted for the mature mouse and human peptides [32]. UT exhibits relatively high sequence identity with somatostatin, opioid and galanin receptors [94]. The urotensinergic system displays an unprecedented repertoire of four or five ancient UT in some vertebrate lineages and five U-II family peptides in teleost fish [91].

Cell penetrating peptides (CPPs) facilitate translocation across biological membranes and are of significant biological and medical interest. Several CPPs can permeate into specific cells and organelles. We examine the incorporation and translocation of a novel anti-cancer CPP in a dioleoylphosphatidylcholine (DOPC) lipid bilayer membrane. The peptide, NAF-1(44-67), is a short fragment of a transmembrane protein, consisting of hydrophobic N terminal and charged C terminal segments. Experiments using fluorescently-labeled NAF-1(44-67) in ~100 nm DOPC vesicles and atomically detailed simulations conducted with Milestoning support a model in which a significant barrier for peptide-membrane entry is found at the interface between the aqueous solution and membrane. The initial step is the insertion of the N terminal segment and the hydrophobic helix into the membrane, passing the hydrophilic head groups. Both experiments and simulations suggest that the free energy difference in the first step of the permeation mechanism in which the hydrophobic helix crosses the phospholipid head groups is −0.4 kcal mol(−1) slightly favoring motion into the membrane. Milestoning calculations of the mean first passage time and the committor function underscore the existence of an early polar barrier followed by a diffusive barrierless motion in the lipid tail region. Permeation events are coupled to membrane fluctuations that are examined in detail. Our study opens the way to investigate in atomistic resolution the molecular mechanism, kinetics, and thermodynamics of CPP permeation to diverse membranes.

The liver performs important functions that are essential for life. If the liver fails, patients will die unless they receive a new liver from a donor (transplant). Unfortunately, there are not enough livers for everyone and some patients die while waiting for a suitable organ. This article describes a novel technique that allows resuscitation and testing of a potential donor liver so that more patients can safely receive a transplant.

We investigate positive subtropical low cloud feedback mechanisms in climate models which have performed the CMIP6/CFMIP-3 AMIP and AMIP uniform +4K experiments while saving CFMIP-3 process diagnostics on model levels. Our analysis focuses on the trade cumulus/stratocumulus transition region between California and Hawaii, where positive low cloud feedbacks are present in the JJA season. We introduce a methodology to} test various positive cloud feedback mechanisms proposed in the literature as primary explanations for the low cloud responses in the models. Causal hypotheses are tested by comparing their predictions with the models’ responses of clouds, cloud controlling factors, boundary layer depth and temperature/humidity tendencies to climate warming. Changes in boundary layer depth, relative humidity in the cloud layer and humidity advection at the top of the boundary layer are shown to distinguish among the hypotheses considered. For the cases examined, our approach rules out 4/5 of the mechanisms considered in half of the models and 3/5 in the remainder. We argue that unambiguously identifying the positive feedback mechanisms operating in models will in some cases require intervention experiments designed to test specific hypotheses.

Ovarian cancer associated ascites contains soluble inhibitory factors. The ascites was cleared of cellular debris by centrifugation as previously described (18). (A, B) The ascites was filtered 0.2 micron filter and fractionated- f1: 50. (C) The ascites was treated with 50 μg/ml proteinase K at 37{degree sign}C for 1 hour and then incubated at 95{degree sign}C for 10min to heat inactivate the enzyme. The antigen presenting cells were treated with the clarified supernantants for 4 h at 37{degree sign}C, then washed extensively with PBS, and cocultured with NKT hybridomas for 20-24 h at 37{degree sign}C. Cytokine released (IL-2) was measured as an indication of NKT cell activation and was measured by standard sandwich ELISA.

Methodology Details

Purpose: Metastatic castration-resistant prostate cancer (mCRPC) is a heterogeneous disease for which better prognostic models for survival are needed. We examined the added value of circulating tumor cell (CTC) enumeration relative to common prognostic laboratory measures from patients with CRPC.Methods: Utility of CTC enumeration as a baseline and postbaseline prognostic biomarker was examined using data from two prospective randomized registration-directed trials (COU-AA-301 and ELM-PC4) within statistical models used to estimate risk for survival. Discrimination and calibration were used to measure model predictive accuracy and the added value for CTC enumeration in the context of a Cox model containing albumin, lactate dehydrogenase (LDH), PSA, hemoglobin, and alkaline phosphatase (ALK). Discrimination quantifies how accurately a risk model predicts short-term versus long-term survivors. Calibration measures the closeness of actual survival time to the predicted survival time.Results: Adding CTC enumeration to a model containing albumin, LDH, PSA, hemoglobin, and ALK (“ALPHA”) improved its discriminatory power. The weighted c-index for ALPHA without CTCs was 0.72 (SE, 0.02) versus 0.75 (SE, 0.02) for ALPHA + CTCs. The increase in discrimination was restricted to the lower-risk cohort. In terms of calibration, adding CTCs produced a more accurate model-based prediction of patient survival. The absolute prediction error for ALPHA was 3.95 months (SE, 0.28) versus 3.75 months (SE, 0.22) for ALPHA + CTCs.Conclusions: Addition of CTC enumeration to standard measures provides more accurate assessment of patient risk in terms of baseline and postbaseline prognosis in the mCRPC population. Clin Cancer Res; 23(8); 1967–73. ©2016 AACR.

The source of GD3 impacts CD1d-mediated antigen presentation.(A) LCD1d cells were treated with GD3 from different sources for 4 hours, then washed extensively and cocultured with NKT cell hybridomas. IL-2 was measured, as an indication of NKT cell activation, by standard cytokine ELISA. (B) Pretreatment of a-GalCer loaded aAPC with GD3. a-GalCer loaded aAPC were incubated for 4hr with either medium, GD3, GD2, or ovarian cancer-associated ascites. The aAPC were washed extensively and then cocultured with a Va14+ NKT cell hybridoma, N38-3C3. NKT cell recognition of CD1d was assessed by measuring IL-2 production in the supernatants by ELISA.

Purpose: The androgen receptor pathway remains active in men with prostate cancer whose disease has progressed following surgical or medical castration. Orteronel (TAK-700) is an investigational, oral, nonsteroidal, selective, reversible inhibitor of 17,20-lyase, a key enzyme in the production of androgenic hormones.Experimental Design: We conducted a phase I/II study in men with progressive, chemotherapy-naïve, metastatic castration-resistant prostate cancer, and serum testosterone Results: In phase I (n = 26), no dose-limiting toxicities were observed and 13 of 20 evaluable patients (65%) achieved ≥50% prostate-specific antigen (PSA) decline from baseline at 12 weeks. In phase II (n = 97), 45 of 84 evaluable patients (54%) achieved a ≥50% decline in PSA and at 12 weeks, substantial mean reductions from baseline in testosterone (−7.5 ng/dL) and dehydroepiandrosterone-sulfate (−45.3 μg/dL) were observed. Unconfirmed partial responses were reported in 10 of 51 evaluable phase II patients (20%). Decreases in circulating tumor cells were documented. Fifty-three percent of phase II patients experienced grade ≥3 adverse events irrespective of causality; most common were fatigue, hypokalemia, hyperglycemia, and diarrhea.Conclusions: 17,20-Lyase inhibition by orteronel was tolerable and results in declines in PSA and testosterone, with evidence of radiographic responses. Clin Cancer Res; 20(5); 1335–44. ©2014 AACR.

PDF file - 65K, Schedule of assessments.

PDF file - 225K, Schematic of Ganglioside GD3 and other glycolipids used or referred to in this paper

Low cloud feedback in global warming projections by climate models is characterized by its positive sign, the mechanism of which is not well understood. Here we propose that the positive sign is primarily caused by the increase in upward longwave radiation from the sea surface. We devise numerical experiments that enable separation of the feedback into components coming from physically distinct causes. Results of these experiments with a climate model indicate that increases in upward longwave radiation from the sea surface cause warming and absolute drying in the boundary layer, leading to the positive low cloud feedback. The absolute drying results from decrease in surface evaporation, and also from decrease in inversion strength which enhances vertical mixing of drier free tropospheric air into the boundary layer. This mechanism is different from previously proposed understanding that positive low cloud feedback is caused by increases in surface evaporation or vertical moisture contrast.

We compare the radiative feedbacks resulting from a uniform warming and cooling of sea surface temperatures by 4 K in an ensemble of global climate models. The global-mean net feedback is less stabilising in response to warming in all nine models. This is primarily due to a stronger tropical water vapour feedback, with a smaller contribution from the shortwave cloud feedback. The zonal-mean feedbacks are similarly robust across the ensemble. In the extra-tropics, more positive shortwave cloud feedback under warming is associated with further poleward migration of the mean Southern Hemisphere jet latitude in some models. However, additional experiments with an aquaplanet version of the HadGEM3 model suggest that the asymmetry of the jet shift is not driving that in the cloud feedbacks at these latitudes. In the tropics, stronger water vapour feedback under warming is offset by a weaker shortwave cloud feedback. The result is that the ensemble spread in the differences between the global feedbacks under warming and cooling is mainly determined by their differences in the tropics. The spatial distribution of the feedbacks largely reflects the zonal mean behaviour, although there is considerable intermodel variation in the regional cloud feedbacks, particularly in the tropical shortwave cloud feedback. Comparison with CO2- and solar-forced coupled experiments suggests that the global-mean longwave cloud feedback is nearly invariant to warming and cooling, regardless of the nature of the forcing. The shortwave cloud feedback is generally more positive under warming in the coupled models, consistent with the uniform SST perturbation experiments.

BackgroundGuidelines emphasize rapid antibiotic treatment for sepsis, but infection presence is often uncertain at initial presentation. We investigated the incidence and drivers of false-positive presumptive infection diagnosis among emergency department (ED) patients meeting Sepsis-3 criteria.MethodsFor a retrospective cohort of patients hospitalized after meeting Sepsis-3 criteria (acute organ failure and suspected infection including blood cultures drawn and intravenous antimicrobials administered) in 1 of 4 EDs from 2013 to 2017, trained reviewers first identified the ED-diagnosed source of infection and adjudicated the presence and source of infection on final assessment. Reviewers subsequently adjudicated final infection probability for a randomly selected 10% subset of subjects. Risk factors for false-positive infection diagnosis and its association with 30-day mortality were evaluated using multivariable regression.ResultsOf 8267 patients meeting Sepsis-3 criteria in the ED, 699 (8.5%) did not have an infection on final adjudication and 1488 (18.0%) patients with confirmed infections had a different source of infection diagnosed in the ED versus final adjudication (ie, initial/final source diagnosis discordance). Among the subset of patients whose final infection probability was adjudicated (n = 812), 79 (9.7%) had only “possible” infection and 77 (9.5%) were not infected. Factors associated with false-positive infection diagnosis included hypothermia, altered mental status, comorbidity burden, and an “unknown infection source” diagnosis in the ED (odds ratio: 6.39; 95% confidence interval: 5.14–7.94). False-positive infection diagnosis was not associated with 30-day mortality.ConclusionsIn this large multihospital study

Introduction: Direct-acting oral anticoagulants (DOACs) and warfarin are standard of care for secondary prevention of ischemic stroke (IS) in persons with atrial fibrillation (AF). However, despite adequate anticoagulation (AC), patients may still experience recurrent IS. We aimed to study the characteristics of patients who developed IS despite therapeutic AC, specifically with DOACs. Methods: Between 2012 and 2021, patients admitted at our hospital with IS despite adequate AC for AF were enrolled. Clinical variables collected included National Institute of Health Stroke Scale (NIHSS), CHA 2 DS 2 -VASc and modified Rankin Score (mRS) at admission. Baseline characteristics were compared between patients taking DOACs and warfarin. Logistic regression analyses were performed to identify predictors of IS in patients using DOACs. Results: A total of 595 patients were included. Of these, 291 patients (48.9%) were on DOACs with a mean age of 75.21 and 43.9% were female. The median CHA 2 DS 2 -VASc and NIHSS score in the DOAC group was 4 and 6, respectively. Compared to those on Warfarin, patients on DOACs were younger (75.21 vs. 78.99 p < 0.001), more likely to be men (55.3% vs. 45.2% p = 0.01) and had higher rates of prior TIA/IS (37.5% vs. 29.3% p = 0.04). Both groups had similar rates of hemorrhagic transformation, but more patients in the DOAC group were on antiplatelet agents concomitantly (21% vs. 6.9% p < 0.001). The most prescribed DOAC was apixaban (66.3%) and cardioembolism was the most common stroke etiology (63.6%), followed by cryptogenic, which was significantly higher in the DOAC group (18.2% vs. 8.3% p = 0.02). Notably, DOAC failure in preventing IS was associated with younger age, overweight/obesity, and prior TIA/IS (Table) . Conclusions: IS despite the use of DOACs is more likely to occur in younger people with prior TIA/IS and obesity. Focusing on preventative strategies and risk factor control for competing mechanisms in this subgroup may help prevent IS recurrence.

Mycoviruses may influence the pathogenicity of disease-causing fungi. Although mycoviruses have been found in some chytrid fungi, limited testing has not detected them in Batrachochytrium dendrobatidis (Bd), the cause of the devastating amphibian disease, chytridiomycosis. Here we conducted a survey for mycovirus presence in 38 Bd isolates from Australia (n = 31), Brazil (n = 5) and South Korea (n = 2) with a combination of modern high-throughput sequencing and conventional dsRNA cellulose chromatography. Mycoviruses were not detected in any isolates. This result was unexpected, given the long evolutionary history of Bd, as well as the high prevalence of mycoviruses in related fungal species. Given our widespread sampling in Australia and the limited number of Bd introductions, we suggest that mycoviruses are uncommon or absent from Australian Bd. Testing more isolates from regions where Bd originated, as well as regions with high diversity or low fungal virulence may identify mycoviruses that could aid in disease control.