Increased COVID-19 mortality among immigrants compared with US-born individuals: a cross-sectional analysis of 2020 mortality data.

OBJECTIVE: Multiple studies have shown that racially minoritized groups had disproportionate COVID-19 mortality relative to non-Hispanic White individuals. However, there is little known regarding mortality by immigrant status nationally in the United States, despite being another vulnerable population. STUDY DESIGN: This was an observational cross-sectional study using mortality vital statistics system data to calculate proportionate mortality ratios (PMRs) and mortality rates due to COVID-19 as the underlying cause. METHODS: Rates were compared by decedents' identified race, ethnicity (Hispanic vs non-Hispanic), and immigrant (immigrants vs US born) status. Asian race was further disaggregated into "Asian Indian," "Chinese," "Filipino," "Japanese," "Korean," and "Vietnamese." RESULTS: Of the over 3.4 million people who died in 2020, 10.4% of all deaths were attributed to COVID-19 as the underlying cause (n = 351,530). More than double (18.9%, n = 81,815) the percentage of immigrants who died of COVID-19 compared with US-born decedents (9.1%, n = 269,715). PMRs due to COVID-19 were higher among immigrants compared with US-born individuals for non-Hispanic White, non-Hispanic Black, Hispanic, and most disaggregated Asian groups. Among disaggregated Asian immigrants, age- and sex-adjusted PMR due to COVID-19 ranged from 1.58 times greater mortality among Filipino immigrants (95% confidence interval [CI]: 1.53, 1.64) to 0.77 times greater mortality among Japanese immigrants (95% CI: 0.68, 0.86). Age-adjusted mortality rates were also higher among immigrant individuals compared with US-born people. CONCLUSIONS: Immigrant individuals experienced greater mortality due to COVID-19 compared with their US-born counterparts. As COVID-19 becomes more endemic, greater clinical and public health efforts are needed to reduce disparities in mortality among immigrants compared with their US-born counterparts.

Hand2 represses non-cardiac cell fates through chromatin remodeling at cis- regulatory elements.

Developmental studies have revealed the importance of the transcription factor Hand2 in cardiac development. Hand2 promotes cardiac progenitor differentiation and epithelial maturation, while repressing other tissue types. The mechanisms underlying the promotion of cardiac fates are far better understood than those underlying the repression of alternative fates. Here, we assess Hand2-dependent changes in gene expression and chromatin remodeling in cardiac progenitors of zebrafish embryos. Cell-type specific transcriptome analysis shows a dual function for Hand2 in activation of cardiac differentiation genes and repression of pronephric pathways. We identify functional cis- regulatory elements whose chromatin accessibility are increased in hand2 mutant cells. These regulatory elements associate with non-cardiac gene expression, and drive reporter gene expression in tissues associated with Hand2-repressed genes. We find that functional Hand2 is sufficient to reduce non-cardiac reporter expression in cardiac lineages. Taken together, our data support a model of Hand2-dependent coordination of transcriptional programs, not only through transcriptional activation of cardiac and epithelial maturation genes, but also through repressive chromatin remodeling at the DNA regulatory elements of non-cardiac genes.

The persistence of emamectin benzoate in marine sediments with different organic matter regimes, temperature conditions, and antibiotic presence.

The objectives of this 318-day study are to determine half-lives of the anti-sea lice medication emamectin benzoate (EMB) under conditions present in sediments at aquaculture sites and document the degradation of EMB into its main metabolite desmethyl emamectin benzoate (DES). Tested conditions include different matrix types (sand, mud), two temperatures (4, 10 degrees), organic matter presence (fish feed waste and feces), and the presence of oxytetracycline. We document a transformation ratio of EMB to DES of 0.16 to 4.4 % and show that the co-presence of oxytetracycline increases EMB calculated half-lives to values >6000 days for mud matrices. EMB incubated in high organic enrichment regimes was not observed to degrade at 4 degrees. Multivariate analyses show interactions between sediment conditions (matrix, temperature, organic matter [OM], oxytetracycline) influence EMB persistence and DES:EMB ratios. Ranges of EMB half-lives and information on metabolites can be used to anticipate potential effects on marine communities.

River Basin Export Reduction Optimization Support Tool; a tool to screen options for reducing nutrient loads while minimizing cost.

Excess loading of nitrogen and phosphorus to river networks causes environmental harm, but reducing loads from large river basins is difficult and expensive. We develop a new tool, the River Basin Export Reduction Optimization Support Tool (RBEROST) to identify least-cost combinations of management practices that will reduce nutrient loading to target levels in downstream and mid-network waterbodies. We demonstrate the utility of the tool in a case study in the Upper Connecticut River basin in New England, USA. The total project cost of optimized lowest-cost plans ranged from $18.0 million to $41.0 million per year over 15 years depending on user specifications. Plans include both point source and non-point source management practices, and most costs are associated with urban stormwater practices. Adding a 2% margin of safety to loading targets improved estimated probability of success from 37.5% to 99%. The large spatial scale of RBEROST, and the consideration of both point and non-point source contributions of nutrients, makes it well suited as an initial screening tool in watershed planning.

Effects of 5-HT2C receptor stimulation in male mice on behaviour and Fos expression: Feeding, reward and impulsivity.

Serotonin modulates many motivated behaviours via multiple receptor subtypes. Agonists at 5-HT2C receptors have potential for treating behavioural problems associated with obesity and drug use. In this work we examined the impact of the 5-HT2C receptor agonist lorcaserin on several motivated behaviours related to feeding, reward and waiting impulsivity, and on neuronal activation in key brain areas mediating those behaviours. In male C57BL/6J mice effects of lorcaserin (0.2, 1 and 5 mg/kg) were examined on feeding, and on operant responding for a palatable reward. Feeding was reduced only at 5 mg/kg, whereas operant responding was reduced at 1 mg/kg. At a much lower dose range lorcaserin 0.05-0.2 mg/kg also reduced impulsive behaviour measured as premature responding in the 5-choice serial reaction time (5-CSRT) test, without affecting attention or ability to perform the task. Lorcaserin induced Fos expression in brain regions related to feeding (paraventricular nucleus and arcuate nucleus), reward (ventral tegmental area), and impulsivity (medial prefrontal cortex, VTA) although these effects did not show the same differential sensitivity to lorcaserin as the behavioural measures. These results indicate a broad profile of action of 5-HT2C receptor stimulation on brain circuitry and on motivated behaviours, but with clear evidence of differential sensitivity across behavioural domains. This is exemplified by the fact that impulsive behaviour was reduced at a much lower dose range than was feeding behaviour. Along with previous work, and some clinical observations, this work supports the idea that 5-HT2C agonists may be useful for behavioural problems associated with impulsivity.

HyU: Hybrid Unmixing for longitudinal in vivo imaging of low signal-to-noise fluorescence.

The expansion of fluorescence bioimaging toward more complex systems and geometries requires analytical tools capable of spanning widely varying timescales and length scales, cleanly separating multiple fluorescent labels and distinguishing these labels from background autofluorescence. Here we meet these challenging objectives for multispectral fluorescence microscopy, combining hyperspectral phasors and linear unmixing to create Hybrid Unmixing (HyU). HyU is efficient and robust, capable of quantitative signal separation even at low illumination levels. In dynamic imaging of developing zebrafish embryos and in mouse tissue, HyU was able to cleanly and efficiently unmix multiple fluorescent labels, even in demanding volumetric timelapse imaging settings. HyU permits high dynamic range imaging, allowing simultaneous imaging of bright exogenous labels and dim endogenous labels. This enables coincident studies of tagged components, cellular behaviors and cellular metabolism within the same specimen, providing more accurate insights into the orchestrated complexity of biological systems.

Metabolization of emamectin benzoate into desmethyl emamectin benzoate in spiked marine sediments.

Emamectin benzoate (EMB) (4″deoxy- 4″-epi-methylaminoavermectin) is a pesticide developed to control pests on various crops, and in forestry. It is also used in salmon aquaculture to control sea lice infestations as an in-feed therapeutant. Little is known about EMB metabolites and potential associated toxicities in marine sediments. In this study, we used natural marine sediments (sand and mud) fortified at an EMB concentration of 60 ppb (wet weight). Results show an almost immediate transformation of EMB to Desmethyl EMB (DES) with no increased rates of metabolization when stored sediment samples were incubated for up to 16 h. The transformation ratio of EMB to DES represented between 0.16 and 0.39% of EMB; values are lower than what has been observed in fish tissue. Data suggest that DES is generated through both abiotic (tested after autoclaving marine sediments) and biological processes. Further work on freshly sampled marine sediments with EMB deposits, different organic regimes, and a detailed assessment of active bacterial communities are necessary to better evaluate the EMB to DES rate of transformation around aquaculture sites.

Implications of tropical cyclones on damage and potential recovery and restoration of logged forests in Vietnam.

Many natural forests in Southeast Asia are degraded following decades of logging. Restoration of these forests is delayed by ongoing logging and tropical cyclones, but the implications for recovery are largely uncertain. We analysed meteorological, satellite and forest inventory plot data to assess the effect of Typhoon Doksuri, a major tropical cyclone, on the forest landscapes of central Vietnam consisting of natural forests and plantations. We estimated the return period for a cyclone of this intensity to be 40 years. Plantations were almost twice as likely to suffer cyclone damage compared to natural forests. Logged natural forests (9-12 years after cessation of government-licensed logging) were surveyed before and after the storm with 2 years between measurements and remained a small biomass carbon sink (0.1 ± 0.3 Mg C ha-1 yr-1) over this period. The cyclone reduced the carbon sink of recovering natural forests by an average of 0.85 Mg C ha-1 yr-1, less than the carbon loss due to ongoing unlicensed logging. Restoration of forest landscapes in Southeast Asia requires a reduction in unlicensed logging and prevention of further conversion of degraded natural forests to plantations, particularly in landscapes prone to tropical cyclones where natural forests provide a resilient carbon sink. This article is part of the theme issue 'Understanding forest landscape restoration: reinforcing scientific foundations for the UN Decade on Ecosystem Restoration'.

DPSC-Derived Extracellular Vesicles Promote Rat Jawbone Regeneration.

Repair and functional reconstruction of large jawbone defects remain one of the challenges in the field of head and neck surgery. The recent progress in tissue engineering technologies and stem cell biology has significantly promoted the development of regenerative reconstruction of jawbone defects. The multiple trophic activities of extracellular vesicles (EVs) produced by mesenchymal stem cells (MSCs) may play a critical role in their therapeutic effects. Accumulating evidence has shown the promise of dental pulp stem cells (DPSCs) in bone regeneration, but less is known about the regenerative effects of DPSC-EVs on jawbone defects. The purpose of this study is to explore the osteogenic effects of DPSC-EVs on jawbone marrow-derived MSCs (JB-MSCs) in vitro and their osteoinductive effects in a mandibular bone defect model in rats. Our results showed that JB-MSCs could efficiently uptake DPSC-EVs, which in turn significantly promoted the expression of osteogenic genes, such as runt-related transcription factor 2 (RUNX2), alkaline phosphatase (ALP), and osteocalcin (OCN), as well as the osteogenic differentiation capability of JB-MSCs. Meanwhile, we found that the pro-osteogenic effect in vitro induced by DPSC-EVs was comparable to that induced by BMP-2 (bone morphogenetic protein 2), currently the only Food and Drug Administration-approved osteoinductive growth factor. In vivo, animals that were locally treated with DPSC-EVs laden with a commercially available collagen membrane exhibited a relatively fast wound closure and increased new bone density at the mandible defects. Our results provide evidence for the osteogenic and osteoinductive effects of DPSC-EVs on jawbone regeneration. Due to the accessibility, rapid proliferation, and osteogenic propensity of DPSCs, DPSC-EVs may represent a safe cell-free therapeutic approach for craniofacial bone regeneration.

FRaeppli: a multispectral imaging toolbox for cell tracing and dense tissue analysis in zebrafish.

Visualizing cell shapes and interactions of differentiating cells is instrumental for understanding organ development and repair. Across species, strategies for stochastic multicolour labelling have greatly facilitated in vivo cell tracking and mapping neuronal connectivity. Yet integrating multi-fluorophore information into the context of developing zebrafish tissues is challenging given their cytoplasmic localization and spectral incompatibility with common fluorescent markers. Inspired by Drosophila Raeppli, we developed FRaeppli (Fish-Raeppli) by expressing bright membrane- or nuclear-targeted fluorescent proteins for efficient cell shape analysis and tracking. High spatiotemporal activation flexibility is provided by the Gal4/UAS system together with Cre/lox and/or PhiC31 integrase. The distinct spectra of the FRaeppli fluorescent proteins allow simultaneous imaging with GFP and infrared subcellular reporters or tissue landmarks. We demonstrate the suitability of FRaeppli for live imaging of complex internal organs, such as the liver, and have tailored hyperspectral protocols for time-efficient acquisition. Combining FRaeppli with polarity markers revealed previously unknown canalicular topologies between differentiating hepatocytes, reminiscent of the mammalian liver, suggesting common developmental mechanisms. The multispectral FRaeppli toolbox thus enables the comprehensive analysis of intricate cellular morphologies, topologies and lineages at single-cell resolution in zebrafish.

Novel human-derived EML4-ALK fusion cell lines identify ribonucleotide reductase RRM2 as a target of activated ALK in NSCLC.

INTRODUCTION: Echinoderm microtubule-associated protein-like 4 (EML4)-Anaplastic Lymphoma Kinase (ALK) rearrangements occur in 3% to 7% of lung adenocarcinomas and are targets for treatment with tyrosine kinase inhibitors (TKIs). Here we have developed three novel EML4-ALK-positive patient-derived Non-Small-Cell-Lung-Cancer (NSCLC) cancer cell lines, CUTO8 (variant 1), CUTO9 (variant 1) and CUTO29 (variant 3) and included a fourth ALK-positive cell line YU1077 (variant 3) to study ALK-positive signaling and responses. Variants 1 and 3 are the most common EML4-ALK variants expressed in ALK-positive NSCLC, and currently cell lines representing these EML4-ALK variants are limited. MATERIALS AND METHODS: Resazurin assay was performed to evaluate cell viability. Protein levels were determined using western blotting. RNA sequencing was performed in all four cell lines to identify differentially expressed genes. Whole-genome sequencing was performed to determine the presence of EML4-ALK fusion and ALK tyrosine kinase inhibitor resistance mutations. RESULTS: In this study, we have confirmed expression of the corresponding ALK fusion protein and assessed their sensitivity to a range of ALK tyrosine kinase inhibitors. These patient derived cell lines exhibit differential sensitivity to lorlatinib, brigatinib and alectinib, with EML4-ALK variant 3 containing cell lines exhibiting increased sensitivity to lorlatinib and brigatinib as compared to alectinib. These cell lines were further characterized by whole genome sequencing and RNA-seq analysis that identified the ribonucleotide reductase regulatory subunit 2 (RRM2) as a downstream and potential therapeutic target in ALK-positive NSCLC. CONCLUSION: We provide a characterization of four novel EML4-ALK-positive NSCLC cell lines, highlighting genomic heterogeneity and differential responses to ALK TKI treatment. The RNA-Seq characterization of ALK-positive NSCLC CUTO8, CUTO9, CUTO29 and YU1077 cell lines reported here, has been compiled in an interactive ShinyApp resource for public data exploration (https://ccgg.ugent.be/shiny/nsclc_rrm2_2022/).

Role of alpha-2 adrenergic and kappa opioid receptors in the effects of alcohol gavage-induced dependence on alcohol seeking.

Noradrenaline and alpha-2 receptors are implicated in the neuroadaptive changes in alcohol dependence leading to increased alcohol seeking. Preclinical methods often used to induce dependence, such as alcohol vapor, require long exposure periods. Another method, gavage with alcohol, induces dependence in a shorter time frame (4-6 d), but its effects on relapse are less well understood. We examined the role of alpha-2 receptors in alcohol self-administration (ASA) and relapse in male and female rats made alcohol dependent by gavage. The influence of these variables on the inhibitory effects of the alpha-2 agonist guanfacine on ASA, and on reinstatement induced by the alpha-2 antagonist yohimbine were determined. We also extended this analysis to relapse induced by the kappa opioid receptor agonist U50,488. Male and female Sprague Dawley rats, trained to self-administer alcohol were treated with intragastric vehicle or alcohol (12 g/kg/d for 5 d). In Exp. 1 we examined the effects of alcohol gavage on reinstatement induced by yohimbine (0.625-1.25 mg/kg) and U50,488 (1.25-2.5 mg/kg). In Exp. 2 we determined the effects of a longer period of alcohol gavage on guanfacine (0.25-0.75 mg/kg)-induced reductions in ASA and on yohimbine (0.625-2.5 mg/kg)-induced reinstatement. Our key findings are that alcohol dependence induced by gavage produces sex-specific effects on reinstatement. Non-dependent females had greater reinstatement than males, but dependence reduced reinstatement in females. In males, dependence modestly enhanced yohimbine-induced reinstatement, while U50-induced reinstatement was absent irrespective of dependence condition. Alcohol dependence did not modify the inhibitory effects of guanfacine on ASA in males or females.

Effects of pimavanserin and lorcaserin on alcohol self-administration and reinstatement in male and female rats.

The neurotransmitter serotonin (5-hydroxytryptamine; 5-HT) modulates fundamental motivational processes, and the neurochemical and behavioural effects of drugs of abuse. Recently, attention has focused on the role of 5-HT acting via 5-HT2A and 5-HT2C receptor sub-types in this regard. We examined the impact of manipulating 5-HT2A and 5-HT2C receptor mediated function on several aspects of alcohol self-administration and alcohol-seeking behaviour in male and female rats. Specifically, experiments investigated the effect of the 5-HT2A inverse agonist/antagonist pimavanserin, and the 5-HT2C receptor agonist lorcaserin on these behaviours. In male and female rats trained to respond for alcohol reinforcement on fixed ratio (FR) and progressive ratio (PR) schedules of reinforcement pimavanserin (0.3, 1 and 3 mg/kg) had no consistent effect on responding. Lorcaserin (0.25, 0.5 and 1 mg/kg) reduced these behaviours in both sexes. Following extinction of responding for alcohol, alcohol-seeking was reinstated by cues previously paired with alcohol. Pimavanserin (1 mg/kg) and lorcaserin (0.5 mg/kg) significantly reduced this reinstatement. In a two-bottle 24 h intermittent access procedure pimavanserin had no significant effects, but lorcaserin reduced alcohol consumption in both sexes at 1, 4 and 24 h after access to alcohol was allowed. Finally, as determined using in vivo microdialysis, alcohol increased, and lorcaserin (0.5 mg/kg) reduced, extracellular levels of DA in the NAc in male rats. In rats treated with lorcaserin prior to alcohol injection the net effect was that DA levels were not changed compared to those measured in control rats. These results suggest that blocking 5-HT2A receptor activity has a very limited action to reduce alcohol-seeking. Activating 5-HT2C receptors had a broader behavioural profile to reduce alcohol self-administration, alcohol drinking and alcohol seeking. These effects may partly result from a blunting of the effect of alcohol on mesolimbic DA release.

Cezanne is a critical regulator of pathological arterial remodelling by targeting ß-catenin signalling.

AIMS: Pathological arterial remodelling including neointimal hyperplasia and atherosclerosis is the main underlying cause for occluding arterial diseases. Cezanne is a novel deubiquitinating enzyme, functioning as a NF-кB negative regulator, and plays a key role in renal inflammatory response and kidney injury induced by ischaemia. Here we attempted to examine its pathological role in vascular smooth muscle cell (VSMC) pathology and arterial remodelling. METHODS AND RESULTS: Cezanne expression levels were consistently induced by various atherogenic stimuli in VSMCs, and in remodelled arteries upon injury. Functionally, VSMCs over-expressing wild-type Cezanne, but not the mutated catalytically-inactive Cezanne (C209S), had an increased proliferative ability and mobility, while the opposite was observed in VSMCs with Cezanne knockdown. Surprisingly, we observed no significant effects of Cezanne on VSMC apoptosis, NF-κB signalling, or inflammation. RNA-sequencing and biochemical studies showed that Cezanne drives VSMC proliferation by regulating CCN family member 1 (CCN1) by targeting ß-catenin for deubiquitination. Importantly, local correction of Cezanne expression in the injured arteries greatly decreased VSMC proliferation, and prevented arterial inward remodelling. Interestingly, global Cezanne gene deletion in mice led to smaller atherosclerotic plaques, but with a lower level of plaque stability. Translating, we observed a similar role for Cezanne in human VSMCs, and higher expression levels of Cezanne in human atherosclerotic lesions. CONCLUSION: Cezanne is a key regulator of VSMC proliferation and migration in pathological arterial remodelling. Our findings have important implications for therapeutic targeting Cezanne signalling and VSMC pathology in vascular diseases.

Evolution of MET and NRAS gene amplification as acquired resistance mechanisms in EGFR mutant NSCLC.

EGFR mutant non-small cell lung cancer patients' disease demonstrates remarkable responses to EGFR-targeted therapy, but inevitably they succumb to acquired resistance, which can be complex and difficult to treat. Analyzing acquired resistance through broad molecular testing is crucial to understanding the resistance mechanisms and developing new treatment options. We performed diverse clinical testing on a patient with successive stages of acquired resistance, first to an EGFR inhibitor with MET gene amplification and then subsequently to a combination EGFR and MET targeted therapies. A patient-derived cell line obtained at the time of disease progression was used to identify NRAS gene amplification as an additional driver of drug resistance to combination EGFR/MET therapies. Analysis of downstream signaling revealed extracellular signal-related kinase activation that could only be eliminated by trametinib treatment, while Akt activation could be modulated by various combinations of MET, EGFR, and PI3K inhibitors. The combination of an EGFR inhibitor with a MEK inhibitor was identified as a possible treatment option to overcome drug resistance related to NRAS gene amplification.

Erratum: Publisher's Note: "Laser-induced electron diffraction of the ultrafast umbrella motion in ammonia" [Struct. Dyn. 8, 014301 (2021)].

[This corrects the article DOI: 10.1063/4.0000046.].

Laser-induced electron diffraction of the ultrafast umbrella motion in ammonia.

Visualizing molecular transformations in real-time requires a structural retrieval method with Ångström spatial and femtosecond temporal atomic resolution. Imaging of hydrogen-containing molecules additionally requires an imaging method sensitive to the atomic positions of hydrogen nuclei, with most methods possessing relatively low sensitivity to hydrogen scattering. Laser-induced electron diffraction (LIED) is a table-top technique that can image ultrafast structural changes of gas-phase polyatomic molecules with sub-Ångström and femtosecond spatiotemporal resolution together with relatively high sensitivity to hydrogen scattering. Here, we image the umbrella motion of an isolated ammonia molecule (NH3) following its strong-field ionization. Upon ionization of a neutral ammonia molecule, the ammonia cation (NH3 +) undergoes an ultrafast geometrical transformation from a pyramidal ( Φ HNH = 107 ° ) to planar ( Φ HNH = 120 ° ) structure in approximately 8 femtoseconds. Using LIED, we retrieve a near-planar ( Φ HNH = 117 ± 5 ° ) field-dressed NH3 + molecular structure 7.8 - 9.8 femtoseconds after ionization. Our measured field-dressed NH3 + structure is in excellent agreement with our calculated equilibrium field-dressed structure using quantum chemical ab initio calculations.

Molecular structure retrieval directly from laboratory-frame photoelectron spectra in laser-induced electron diffraction.

Ubiquitous to most molecular scattering methods is the challenge to retrieve bond distance and angle from the scattering signals since this requires convergence of pattern matching algorithms or fitting methods. This problem is typically exacerbated when imaging larger molecules or for dynamic systems with little a priori knowledge. Here, we employ laser-induced electron diffraction (LIED) which is a powerful means to determine the precise atomic configuration of an isolated gas-phase molecule with picometre spatial and attosecond temporal precision. We introduce a simple molecular retrieval method, which is based only on the identification of critical points in the oscillating molecular interference scattering signal that is extracted directly from the laboratory-frame photoelectron spectrum. The method is compared with a Fourier-based retrieval method, and we show that both methods correctly retrieve the asymmetrically stretched and bent field-dressed configuration of the asymmetric top molecule carbonyl sulfide (OCS), which is confirmed by our quantum-classical calculations.