Search

Aims The study of microbial inactivation rates in aquifer systems has most often been determined in aerobic and oxidized systems. This study examined the inactivation (i.e. loss of infectivity) of MS2, poliovirus type 1 (PV1) and Cryptosporidium parvum in an anaerobic and reduced groundwater system that has been identified as storage zones for aquifer storage and recovery (ASR) facilities. Methods and Results Anaerobic and reduced (ORP < −250 mV) groundwater from an artesian well was diverted to an above-ground, flow-through mesocosm that contained diffusion chambers filled with MS2, PV1 or Cryptosporidium parvum. The respective infectivity assays were performed on microorganisms recovered from the diffusion chambers during 30- to 58-day experiments. The net reduction in infectivity was 5.73 log10 over 30 days for MS2, 5.00 log10 over 58 days for PV1 and 4.07 log10 over 37 days for C. parvum. The best fit inactivation model for PV1 was the log-linear model and the Weibull model for MS2 and C. parvum, with respective inactivation rates (95% confidence interval) of 0.19 (0.17–0.21) log10 day−1, 0.31 (0.19–0.89) log10 day−1 and 0.20 (0.14–0.37) log10 day−1. Conclusions The groundwater geochemical conditions in this aquifer enhanced the inactivation of MS2, PV1, and C. parvum at rates approximately 2.0–5.3-fold, 1.2–17.0-fold, and 4.5–5.6-fold greater, respectively, than those from published studies that used diffusion chambers in aerobic-to-anoxic groundwater systems, with positive redox potentials. Significance and Impact of the Study Geochemical conditions like those in the aquifer zone in this study can naturally and significantly reduce concentrations of microbial indicators and pathogens of human health concern in injected surface water. Appropriate storage times for injected surface water could complement above-ground engineered processes for microorganism removal and inactivation (e.g. filtration, disinfection) by naturally increasing overall microorganism log-inactivation rates of ASR facilities.

G-Protein coupled D2 autoreceptors expressed on dopamine neurons (D2Rs) inhibit transmitter release and cell firing at axonal endings and somatodendritic compartments. Mechanistic details of somatodendritic dopamine release remain unresolved, partly due to insufficient information on the subcellular distribution of D2Rs. Previous studies localizing D2Rs have been hindered by a dearth of antibodies validated for specificity in D2R knockout animals and have been limited by the small sampling areas imaged by electron microscopy. This study utilized sub-diffraction fluorescence microscopy and electron microscopy to examine D2 receptors in a superecliptic pHlourin GFP (SEP) epitope tagged D2 receptor knockin mouse. Incubating live slices with an anti-SEP antibody achieved the selective labeling of plasma membrane associated receptors for immunofluorescent imaging over a large area of the substantia nigra pars compacta (SNc). SEP-D2Rs appeared as puncta-like structures along the surface of dendrites and soma of dopamine neurons visualized by antibodies to tyrosine hydroxylase (TH). TH-associated SEP-D2Rs displayed a cell surface density of 0.66 puncta per μm(2), which corresponds to an average frequency of 1 punctum every 1.50 μm. Separate ultrastructural experiments using silver-enhanced immunogold revealed that membrane-bound particles represented 28% of total D2Rs in putative dopamine cells within the SNc. Structures immediately adjacent to dendritic membrane gold particles were unmyelinated axons or axon varicosities (40%), astrocytes (19%), other dendrites (7%), or profiles unidentified (34%) in single sections. Some apposed profiles also expressed D2Rs. Fluorescent and ultrastructural analyses also provided the first visualization of membrane-D2Rs at the axon initial segment, a compartment critical for action potential generation. The punctate appearance of anti-SEP staining indicates there is a population of D2Rs organized in discrete signaling sites along the plasma membrane, and for the first time a quantitative estimate of spatial frequency is provided.

Purpose: We performed a NCI-sponsored, prospective study of neoadjuvant FOLFIRINOX followed by chemoradiation with carboplatin/paclitaxel followed by surgery in patients with locally advanced gastric or gastroesophageal cancer. Patients and Methods: The primary objective was to determine completion rate of neoadjuvant FOLFIRINOX × 8 followed by chemoradiation. Secondary endpoints were toxicity and pathologic complete response (pCR) rate. Exploratory analysis was performed of circulating tumor DNA (ctDNA) to treatment response. Results: From October 2017 to June 2018, 25 patients were enrolled. All patients started FOLFIRINOX, 92% completed all eight planned cycles, and 88% completed chemoradiation. Twenty (80%) patients underwent surgical resection, and 7 had a pCR (35% in resected cohort, 28% intention to treat). Tumor-specific mutations were identified in 21 (84%) patients, of whom 4 and 17 patients had undetectable and detectable ctDNA at baseline, respectively. Presence of detectable post-chemoradiation ctDNA (P = 0.004) and/or postoperative ctDNA (P = 0.045) were associated with disease recurrence. Conclusions: Here we show neoadjuvant FOLFIRINOX followed by chemoradiation for locally advanced gastroesophageal cancer is feasible and yields a high rate of pCR. ctDNA appears to be a promising predictor of postoperative recurrence. See related commentary by Catenacci, p. 6281

Photodeoxygenation of dibenzothiophene S-oxide and its derivatives have been used to generate atomic oxygen [O(3P)] to examine its effect on proteins, nucleic acids, and lipids. The unique reactivity and selectivity of O(3P) have shown distinct oxidation products and outcomes in biomolecules and cell-based studies. To understand the scope of its global impact on the cell, we treated MDA-MB-231 cells with 2,8-diacetoxymethyldibenzothiophene S-oxide and UV-A light to produce O(3P) without targeting a specific cell organelle. Cellular responses to O(3P)-release were analyzed using cell viability and cell cycle phase determination assays. Cell death was observed when cells were treated with higher concentrations of sulfoxides and UV-A light. However, significant differences in cell cycle phases due to UV-A irradiation of the sulfoxide were not observed. We further performed RNA-Seq analysis to study the underlying biological processes at play, and while UV-irradiation itself influenced gene expression, there were 9 upregulated and 8 downregulated genes that could be attributed to photodeoxygenation.

Quantitative measurement of process-specific biomarker genes of Dehalococcoides mccartyi (Dhc) supports monitoring at chlorinated ethene contaminated sites. In this study, we varied Dhc cell abunda...

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

A modular synthesis of mycobactin T and its N-acetyl analogue is reported in a route that facilitates permutation of the lipid tails. A key feature is the generation of N(α)-Cbz-N(e)-benzyloxy-N(e)-Boc-lysine (A4) with methyl(trifluoromethyl)dioxirane in 59% yield. Selective hydroxamate N-acylation was achieved with acyl fluorides, enabling installation of lipids tails in the final step. O-Benzyl-dehydrocobactin T (B4) was prepared by modifying a known five-step sequence with an overall yield of 49%. 2-Hydroxyphenyl-4-carboxyloxazoline (C3) was prepared from 2-hydroxybenzoic acid and l-serine methyl ester in three steps with an overall yield of 55%. Ester coupling of A4 and B4 with EDCI afforded MbI-1 in 73% yield. Catalytic hydrogenation with Pd/BaSO4 and 50 psi of H2 simultaneously effected alkene reduction and debenzylation to afford MbI-2 in 96% yield. Fragment C3 was converted into acyl fluoride C4, which coupled with MbI-2 to afford MbI-3 in 51% yield. Finally, Boc-removal with HCl/EtOAc and treatment of the resultant hydroxylamine with stearyl fluoride furnished mycobactin T in 65% yield. Overall, the yield is 4% over 14 steps. The gallium mycobactin T-N-acetyl derivative (GaMbT-NAc) structure was determined by 1H NMR. The structure shows an octahedral Ga and two internal hydrogen bonds between peptidic N-Hs and two of the oxygen atoms coordinating Ga.

Chemical ionization mass spectrometry (CIMS) using iodide as a reagent ion has been widely used to classify organic compounds in the atmosphere by their elemental formula. Unfortunately, calibration of these instruments is challenging due to a lack of commercially available standards for many compounds, which has led to the development of methods for estimating CIMS sensitivity. By coupling a thermal desorption aerosol gas chromatograph (TAG) simultaneously to a flame ionization detector (FID) and an iodide CIMS, we use the individual particle-phase analytes, quantified by the FID, to examine the sensitivity of the CIMS and its variability between isomers of the same elemental formula. Iodide CIMS sensitivities of isomers within a formula are found to generally vary by 1 order of magnitude with a maximum deviation of 2 orders of magnitude. Furthermore, we compare directly measured sensitivity to a method of estimating sensitivity based on declustering voltage (i.e., "voltage scanning"). This approach is found to carry high uncertainties for individual analytes (0.5 to 1 order of magnitude) but represents a central tendency that can be used to estimate the sum of analytes with reasonable error (similar to 30% differences between predicted and measured moles). Finally, gas chromatography (GC) retention time, which is associated with vapor pressure and chemical functionality of an analyte, is found to qualitatively correlate with iodide CIMS sensitivity, but the relationship is not close enough to be quantitatively useful and could be explored further in the future as a potential calibration approach. Alfred P. Sloan FoundationAlfred P. Sloan Foundation [P-2018-11129] Published version This research has been supported by the Alfred P. Sloan Foundation (grant no. P-2018-11129).

Specialized diterpenoid metabolites are important mediators of plant-environment interactions in monocot crops. To understand metabolite functions in plant environmental adaptation that ultimately can enable crop improvement strategies, a deeper knowledge of the underlying species-specific biosynthetic pathways is required. Here, we report the genomics-enabled discovery of five cytochrome P450 monooxygenases (CYP71Z25-CYP71Z29) that form previously unknown furanoditerpenoids in the monocot bioenergy crop Panicum virgatum (switchgrass). Combinatorial pathway reconstruction showed that CYP71Z25-CYP71Z29 catalyze furan ring addition directly to primary diterpene alcohol intermediates derived from distinct class II diterpene synthase products. Transcriptional co-expression patterns and the presence of select diterpenoids in switchgrass roots support the occurrence of P450-derived furanoditerpenoids in planta. Integrating molecular dynamics, structural analysis and targeted mutagenesis identified active site determinants that contribute to the distinct catalytic specificities underlying the broad substrate promiscuity of CYP71Z25-CYP71Z29 for native and non-native diterpenoids.

One-carbon units, critical intermediates for cell growth, may be produced by a variety of means, one of which is via the production of formate. Excessive formate accumulation, known as formate overflow and a characteristic of oxidative cancer, has been observed in cancer cells. However, the basis for this high rate of formate production is unknown. We examined the effect of elevated expression of oncogenic Ras (RasV12), on formate production in NIH-3T3 cells (mouse fibroblasts) cultured with either labelled 13C-serine or 13C-glycine. Formate accumulation by the fibroblasts transformed by RasV12 was increased two-threefold over those by vector control (Babe) cells. The production of formate exceeded the rate of utilization in both cell types. 13C-formate was produced almost exclusively from the #3 carbon of 13C-serine. Virtually no labelled formate was produced from either the #2 carbon of serine or the #2 carbon of glycine. The increased formate production by RasV12 cells was associated with increased mRNA abundances for enzymes of formate production in both the mitochondria and the cytosol. Thus, we find the oncogenic RasV12 significantly increases formate overflow and may be one way for tumor cells to produce one-carbon units required for enhanced proliferation of these cells and/or for other processes which have not been identified.

Myzus persicae has evolved resistance to various insecticides in Greece. Here we examine the effectiveness of the insecticide flupyradifurone against aphid clones collected from tobacco and peach in Greece during 2017-2020. Furthermore, we monitored the frequency of the neonicotinoid resistance mutation R81T in the sampled clones, and the association between the responses to flupyradifurone and acetamiprid.Of 43 clones tested with flupyradifurone, 6.977%, 60.465% and 32.558% showed low (10-14), moderate (19-89) and high (104-1914) resistance factor (RF) values, respectively. Resistance was higher in clones from peach than from tobacco with 42.308% and 17.647% of clones (respectively) failing into the high RF category (median RF values 67.5 and 36.4 for clones from peach and tobacco, respectively). Acetamiprid resistance was detected in clones collected in 2019-2020, in line with our previous study in Greece. The analysis of the whole dataset (54 clones collected during 2017-2020) revealed that all tobacco clones had RF 7.5, whereas 55.263%, 18.421% and 26.316% of the peach clones exhibited low (12), moderate (20-48) and high (100-145) RF values, respectively. A significant but moderate association between flupyradifurone and acetamiprid responses was detected (r = 0.513, P 0.001). The R81T mutation was detected in aphids from peach (5.6% and 32.6% as homozygotes and heterozygotes, respectively) and in one aphid specimen (heterozygote) from tobacco. R81T was partially associated with the resistance to both insecticides, but many highly resistant clones did not possess the mutation, indicating the possible operation of one or more alternative underlying resistance mechanisms.The use of flupyradifurone and acetamiprid in IPM/IRM should be based on further ongoing susceptibility monitoring. © 2021 Society of Chemical Industry.

High-energy batteries for automotive applications require cells to endure well over a decade of constant use, making their long-term stability paramount. This is particularly challenging for emerging cell chemistries containing silicon, for which extended testing information is scarce. While much of the research on silicon anodes has focused on mitigating the consequences of volume changes during cycling, comparatively little is known about the time-dependent degradation of silicon-containing batteries. Here we discuss a series of studies on the reactivity of silicon that, collectively, paint a picture of how the chemistry of silicon exacerbates the calendar aging of lithium-ion cells. Assessing and mitigating this shortcoming should be the focus of future research to fully realize the benefits of this battery technology. Silicon-containing batteries are increasingly becoming a reality in the mass market, but their calendar aging behaviours have received comparatively little attention. Researchers from the Silicon Consortium Project discuss the issues surrounding the calendar lifetime of silicon anodes for lithium-ion batteries.

The global COVID-19 pandemic has created an urgent demand for large numbers of inexpensive, accurate, rapid, point-of-care diagnostic tests. Analyte-based assays are suitably rapid and inexpensive and can be rapidly mass-produced, but for sufficiently accurate performance, they require highly optimized antibodies and assay conditions. We used an automated liquid handling system, customized to handle arrays of lateral flow (immuno)assays (LFAs) in a high-throughput screen, to identify anti-nucleocapsid antibodies that will perform optimally in an LFA. We tested 1021 anti-nucleocapsid antibody pairs as LFA capture and detection reagents with the goal of highlighting pairs that have the greatest affinity for the nucleocapsid protein of SARS-CoV-2 within the LFA format. In contrast to traditional antibody screening methods (e.g., ELISA, bio-layer interferometry), the method described here integrates real-time reaction kinetics with transport in, and immobilization directly onto, nitrocellulose. We have identified several candidate antibody pairs that are suitable for further development of an LFA for SARS-CoV-2.

Inhibition of the bromodomain and extra-terminal (BET) family of adaptor proteins is an attractive strategy for targeting transcriptional regulation of key oncogenes, such as c-MYC. Starting with the screening hit 1, a combination of structure-activity relationship and protein structure-guided drug design led to the discovery of a differently oriented carbazole 9 with favorable binding to the tryptophan, proline, and phenylalanine (WPF) shelf conserved in the BET family. Identification of an additional lipophilic pocket and functional group optimization to optimize pharmacokinetic (PK) properties culminated in the discovery of 18 (BMS-986158) with excellent potency in binding and functional assays. On the basis of its favorable PK profile and robust in vivo activity in a panel of hematologic and solid tumor models, BMS-986158 was selected as a candidate for clinical evaluation.

The Mg0 promoted reduction of a sterically hindered 1,6‐dibromohexasilane yielded a cyclopentasilane as the main product. The disilane t‐BuPhHSi‐SiHPht‐Bu was identified as a minor product, which suggests a possible mechanism involving elimination of a transient silylene SiPht‐Bu.

Heat treatment is one of the most widely used processing technologies in the dairy industry. Its primary purpose is to destroy microorganisms, both pathogenic and spoilage, to ensure the product is safe and has a reasonable shelf life. In this study microwave volumetric heating (MVH) was compared with a conventional tubular heat exchanger (THE), in terms of the effects of each at a range of temperatures (75°C, 85°C, 95°C, 105°C, 115°C, and 125°C) on indigenous microflora viability and the germination of inoculated Bacillus licheniformis endospores in reconstituted skim milk. To assess the heat treatment-related effects on microbial viability, classical agar-based tests were applied to obtain the counts of 4 various microbiological groups including total bacterial, thermophilic bacterial, mesophilic aerobic bacterial endospore, and thermophilic aerobic bacterial endospore counts, and additional novel insights into cell permeability and spore germination profiles post-heat treatment were obtained using real-time flow cytometry (FC) methods. No significant differences in the plate counts of the indigenous microorganisms tested, the plate counts of the inoculated B. licheniformis, or the relative percentage of germinating endospores were observed between MVH- and THE-treated samples, at equal temperatures in the range specified above, indicating that both methods inactivated inoculated endospores to a similar degree (up to 70% as measured by FC and 5 log reduction as measured by plate counting for some treatments of inoculated endospores). Furthermore, increased cell permeability of indigenous microflora was observed by FC after MVH compared with THE treatment of uninoculated skim milk, which was reflected in lower total bacterial count at a treatment temperature of 105°C. This work demonstrates the utility of FC as a rapid method for assessing cell viability and spore inactivation for postthermal processing in dairy products and overall provides evidence that MVH is at least as effective at eliminating native microflora and inoculated B. licheniformis endospores as THE.

CaFe2O4-type sodium postspinels (Na-CFs), with Na+ occupying tunnel sites, are of interest as prospective battery electrodes. While many compounds of this structure type require high-pressure synthesis, several compounds are known to form at ambient pressure. Here we report a large expansion of the known Na-CF phase space at ambient pressure, having successfully synthesized NaCrTiO4, NaRhTiO4, NaCrSnO4, NaInSnO4, NaMg0.5Ti1.5O4, NaFe0.5Ti1.5O4, NaMg0.5Sn1.5O4, NaMn0.5Sn1.5O4, NaFe0.5Sn1.5O4, NaCo0.5Sn1.5O4, NaNi0.5Sn1.5O4, NaCu0.5Sn1.5O4, NaZn0.5Sn1.5O4, NaCd0.5Sn1.5O4, NaSc1.5Sb0.5O4, Na1.16In1.18Sb0.66O4, and several solid solutions. In contrast to earlier reports, even cations that are strongly Jahn-Teller active (e.g., Mn3+ and Cu2+) can form Na-CFs at ambient pressure when combined with Sn4+ rather than with the smaller Ti4+. Order and disorder are probed at the average and local length-scales with synchrotron powder X-ray diffraction and solid-state NMR spectroscopy. Strong ordering of framework cations between the two framework sites is not observed, except in the case of Na1.16In1.18Sb0.66O4. This compound is the first example of an Na-CF that contains Na+ in both the tunnel and framework sites, reminiscent of Li-rich spinels. Trends in the thermodynamic stability of the new compounds are explained on the basis of crystal-chemistry and density functional theory (DFT). Further DFT calculations examine the relative stability of the CF versus spinel structures at various degrees of sodium extraction in the context of electrochemical battery reactions.

The availability of nitrogen (N) in ocean surface waters affects rates of photosynthesis and marine ecosystem structure. In spite of low dissolved inorganic N concentrations, export production in oligotrophic waters is comparable to more nutrient replete regions. Prior observations raise the possibility that di-nitrogen (N2) fixation supplies a significant fraction of N supporting export production in the Gulf of Mexico. In this study, geochemical tools were used to quantify the relative and absolute importance of both subsurface nitrate and N2 fixation as sources of new N fueling export production in the oligotrophic Gulf of Mexico in May 2017 and May 2018. Comparing the isotopic composition (“δ15N”) of nitrate with the δ15N of sinking particulate N collected during five sediment trap deployments each lasting two to four days indicates that N2 fixation is typically not detected and that the majority (≥80%) of export production is supported by subsurface nitrate. Moreover, no gradients in upper ocean dissolved organic N and suspended particulate N concentration and/or δ15N were found that would indicate significant N2 fixation fluxes accumulated in these pools, consistent with low Trichodesmium spp. abundance. Finally, comparing the δ15N of sinking particulate N captured within vs. below the euphotic zone indicates that during late spring regenerated N is low in δ15N compared to sinking N.

Glyphosate is the active ingredient in Roundup® brand nonselective herbicides, and residue testing for food has been conducted as part of the normal regulatory processes. Additional testing has been conducted by university researchers and nongovernmental agencies. Presence of residues needs to be put into the context of safety standards. Furthermore, to appropriately interpret residue data, analytical assays must be validated for each food sample matrix. Regulatory agency surveys indicate that 99% of glyphosate residues in food are below the European maximum residue limits (MRLs) or U.S. Environmental Protection Agency tolerances. These data support the conclusion that overall residues are not elevated above MRLs/tolerances due to agricultural practices or usage on genetically modified (GM) crops. However, it is important to understand that MRLs and tolerances are limits for legal pesticide usage. MRLs only provide health information when the sum of MRLs of all foods is compared to limits established by toxicology studies, such as the acceptable daily intake (ADI). Conclusions from dietary modeling that use actual food residues, or MRLs themselves, combined with consumption data indicate that dietary exposures to glyphosate are within established safe limits. Measurements of glyphosate in urine can also be used to estimate ingested glyphosate exposure, and studies indicate that exposure is

Objective Our objective was to evaluate subsequent feedlot performance and carcass characteristics of steers that previously grazed endophyte-infected fescue and received either a mineral only (MIN) supplement or a cumulative management (CM) strategy. Materials and Methods Grazing occurred in autumn and winter of 2018 and 2019 with feedlot finishing after grazing. Within year, 60 fall-weaned steers (282 ± 7.7 kg) were randomly assigned to either MIN (n = 3) or CM (n = 3) pastures. The CM strategy include a steroidal growth implant, monensin, and 1% BW (as-fed basis) of 1:1 corn gluten feed:soybean hull mix. During feedlot finishing, steers were penned to maintain pasture groups. Results and Discussion Steers receiving CM were heavier (P = 0.002) entering the feedlot and finished 40 d earlier than MIN (161 vs. 201 ± 5.7 d, P Implications and Applications A CM strategy increases weight of steers grazing toxic fescue. This additional weight can reduce the days required for feedlot finishing without influencing carcass characteristics. Improved grazing performance and fewer days for feedlot finishing may reduce environmental impact.

SARS-CoV-2 virus has recently given rise to the current COVID-19 pandemic where infected individuals can range from being asymptomatic, yet highly contagious, to dying from acute respiratory distress syndrome. Although the world has mobilized to create antiviral vaccines and therapeutics to combat the scourge, their long-term efficacy remains in question especially with the emergence of new variants. In this work, we exploit a class of compounds that has previously shown success against various viruses. A salicylanilide library was first screened in a SARS-CoV-2 activity assay in Vero cells. The most efficacious derivative was further evaluated in a prophylactic mouse model of SARS-CoV-2 infection unveiling a salicylanilide that can reduce viral loads, modulate key cytokines, and mitigate severe weight loss involved in COVID-19 infections. The combination of anti-SARS-CoV-2 activity, cytokine inhibitory activity, and a previously established favorable pharmacokinetic profile for the lead salicylanilide renders salicylanilides in general as promising therapeutics for COVID-19.

Aims Emerging evidence suggests that remnant cholesterol (RC) promotes atherosclerotic cardiovascular disease (ASCVD). We aimed to estimate RC-related risk beyond low-density lipoprotein cholesterol (LDL-C) and apolipoprotein B (apoB) in patients without known ASCVD. Methods and results We pooled data from 17 532 ASCVD-free individuals from the Atherosclerosis Risk in Communities study (n = 9748), the Multi-Ethnic Study of Atherosclerosis (n = 3049), and the Coronary Artery Risk Development in Young Adults (n = 4735). RC was calculated as non-high-density lipoprotein cholesterol (non-HDL-C) minus calculated LDL-C. Adjusted Cox models were used to estimate the risk for incident ASCVD associated with log RC levels. We also performed discordance analyses examining relative ASCVD risk in RC vs. LDL-C discordant/concordant groups using difference in percentile units (>10 units) and clinically relevant LDL-C targets. The mean age of participants was 52.3 ± 17.9 years, 56.7% were women and 34% black. There were 2143 ASCVD events over the median follow-up of 18.7 years. After multivariable adjustment including LDL-C and apoB, log RC was associated with higher ASCVD risk [hazard ratio (HR) 1.65, 95% confidence interval (CI) 1.45–1.89]. Moreover, the discordant high RC/low LDL-C group, but not the low RC/high LDL-C group, was associated with increased ASCVD risk compared to the concordant group (HR 1.21, 95% CI 1.08–1.34). Similar results were shown when examining discordance across clinical cutpoints. Conclusions In ASCVD-free individuals, elevated RC levels were associated with ASCVD independent of traditional risk factors, LDL-C, and apoB levels. The mechanisms of RC association with ASCVD, surprisingly beyond apoB, and the potential value of targeted RC-lowering in primary prevention need to be further investigated.

Control of cellular function is extremely complex, being reliant on a wide range of components. Several of these are small oxygen-based molecules. Although reactive compounds containing oxygen are usually harmful to cells when accumulated to relatively high concentrations, they are also instrumental in the control of the activity of a myriad of proteins, and control both the upregulation and downregulation of gene expression. The formation of one oxygen-based molecule, such as the superoxide anion, can lead to a cascade of downstream generation of others, such as hydrogen peroxide (H2O2) and the hydroxyl radical (∙OH), each with their own reactivity and effect. Nitrogen-based signaling molecules also contain oxygen, and include nitric oxide (NO) and peroxynitrite, both instrumental among the suite of cell signaling components. These molecules do not act alone, but form part of a complex interplay of reactions, including with several sulfur-based compounds, such as glutathione and hydrogen sulfide (H2S). Overaccumulation of oxygen-based reactive compounds may alter the redox status of the cell and lead to programmed cell death, in processes referred to as oxidative stress, or nitrosative stress (for nitrogen-based molecules). Here, an overview of the main oxygen-based molecules involved, and the ramifications of their production, is given.

Summary Background PD-1 and PD-L1 inhibitors are active in metastatic urothelial carcinoma, but positive randomised data supporting their use as a first-line treatment are lacking. In this study we assessed outcomes with first-line pembrolizumab alone or combined with chemotherapy versus chemotherapy for patients with previously untreated advanced urothelial carcinoma. Methods KEYNOTE-361 is a randomised, open-label, phase 3 trial of patients aged at least 18 years, with untreated, locally advanced, unresectable, or metastatic urothelial carcinoma, with an Eastern Cooperative Oncology Group performance status of up to 2. Eligible patients were enrolled from 201 medical centres in 21 countries and randomly allocated (1:1:1) via an interactive voice-web response system to intravenous pembrolizumab 200 mg every 3 weeks for a maximum of 35 cycles plus intravenous chemotherapy (gemcitabine [1000 mg/m2] on days 1 and 8 and investigator's choice of cisplatin [70 mg/m2] or carboplatin [area under the curve 5] on day 1 of every 3-week cycle) for a maximum of six cycles, pembrolizumab alone, or chemotherapy alone, stratified by choice of platinum therapy and PD-L1 combined positive score (CPS). Neither patients nor investigators were masked to the treatment assignment or CPS. At protocol-specified final analysis, sequential hypothesis testing began with superiority of pembrolizumab plus chemotherapy versus chemotherapy alone in the total population (all patients randomly allocated to a treatment) for the dual primary endpoints of progression-free survival (p value boundary 0·0019), assessed by masked, independent central review, and overall survival (p value boundary 0·0142), followed by non-inferiority and superiority of overall survival for pembrolizumab versus chemotherapy in the patient population with CPS of at least 10 and in the total population (also a primary endpoint). Safety was assessed in the as-treated population (all patients who received at least one dose of study treatment). This study is completed and is no longer enrolling patients, and is registered at ClinicalTrials.gov , number NCT02853305 . Findings Between Oct 19, 2016 and June 29, 2018, 1010 patients were enrolled and allocated to receive pembrolizumab plus chemotherapy (n=351), pembrolizumab monotherapy (n=307), or chemotherapy alone (n=352). Median follow-up was 31·7 months (IQR 27·7–36·0). Pembrolizumab plus chemotherapy versus chemotherapy did not significantly improve progression-free survival, with a median progression-free survival of 8·3 months (95% CI 7·5–8·5) in the pembrolizumab plus chemotherapy group versus 7·1 months (6·4–7·9) in the chemotherapy group (hazard ratio [HR] 0·78, 95% CI 0·65–0·93; p=0·0033), or overall survival, with a median overall survival of 17·0 months (14·5–19·5) in the pembrolizumab plus chemotherapy group versus 14·3 months (12·3–16·7) in the chemotherapy group (0·86, 0·72–1·02; p=0·0407). No further formal statistical hypothesis testing was done. In analyses of overall survival with pembrolizumab versus chemotherapy (now exploratory based on hierarchical statistical testing), overall survival was similar between these treatment groups, both in the total population (15·6 months [95% CI 12·1–17·9] with pembrolizumab vs 14·3 months [12·3–16·7] with chemotherapy; HR 0·92, 95% CI 0·77–1·11) and the population with CPS of at least 10 (16·1 months [13·6–19·9] with pembrolizumab vs 15·2 months [11·6–23·3] with chemotherapy; 1·01, 0·77–1·32). The most common grade 3 or 4 adverse event attributed to study treatment was anaemia with pembrolizumab plus chemotherapy (104 [30%] of 349 patients) or chemotherapy alone (112 [33%] of 342 patients), and diarrhoea, fatigue, and hyponatraemia (each affecting four [1%] of 302 patients) with pembrolizumab alone. Six (1%) of 1010 patients died due to an adverse event attributed to study treatment; two patients in each treatment group. One each occurred due to cardiac arrest and device-related sepsis in the pembrolizumab plus chemotherapy group, one each due to cardiac failure and malignant neoplasm progression in the pembrolizumab group, and one each due to myocardial infarction and ischaemic colitis in the chemotherapy group. Interpretation The addition of pembrolizumab to first-line platinum-based chemotherapy did not significantly improve efficacy and should not be widely adopted for treatment of advanced urothelial carcinoma. Funding Merck Sharp and Dohme, a subsidiary of Merck, Kenilworth, NJ, USA.

A dopamine D2 receptor mutation was recently identified in a family with a novel hyperkinetic movement disorder. Compared to the wild type D2 receptor, the novel allelic variant D2-I(212)F activates a Gα(i1)β(1)γ(2) heterotrimer with higher potency and modestly enhanced basal activity in human embryonic kidney (HEK) 293 cells, and has decreased capacity to recruit arrestin3. We now report that omitting overexpressed G protein-coupled receptor kinase-2 (GRK2) decreased the potency and efficacy of quinpirole for arrestin recruitment. The relative efficacy of quinpirole for arrestin recruitment to D2-I(212)F compared to D2-WT was considerably lower without overexpressed GRK2 than with added GRK2. D2-I(212)F exhibited higher basal activation of Gα(oA) than Gα(i1), but little or no increase in the potency of quinpirole relative to D2-WT. Other signs of D2-I(212)F constitutive activity for G protein-mediated signaling, in addition to basal activation of Gα(i/o), were enhanced basal inhibition of forskolin-stimulated cyclic AMP accumulation that was reversed by the inverse agonists sulpiride and spiperone and a ~4-fold increase in the apparent affinity of D2-I(212)F for quinpirole, determined from competition binding assays. In mouse midbrain slices, inhibition of tonic current by the inverse agonist sulpiride in dopamine neurons expressing D2-I(212)F was consistent with our hypothesis of enhanced constitutive activity and sensitivity to dopamine relative to D2-WT. Molecular dynamics simulations with D2 receptor models suggested that an ionic lock between the cytoplasmic ends of the third and sixth α-helices that constrains many G protein-coupled receptors in an inactive conformation spontaneously breaks in D2-I(212)F. Overall, these results confirm that D2-I(212)F is a constitutively active and signaling-biased D2 receptor mutant, and also suggest that the effect of the likely pathogenic variant in a given brain region will depend on the nature of G protein and GRK expression.

Metallic zinc is an ideal anode due to its high theoretical capacity (820 mAh g−1), low redox potential (−0.762 V versus the standard hydrogen electrode), high abundance and low toxicity. When used in aqueous electrolyte, it also brings intrinsic safety, but suffers from severe irreversibility. This is best exemplified by low coulombic efficiency, dendrite growth and water consumption. This is thought to be due to severe hydrogen evolution during zinc plating and stripping, hitherto making the in-situ formation of a solid–electrolyte interphase (SEI) impossible. Here, we report an aqueous zinc battery in which a dilute and acidic aqueous electrolyte with an alkylammonium salt additive assists the formation of a robust, Zn2+-conducting and waterproof SEI. The presence of this SEI enables excellent performance: dendrite-free zinc plating/stripping at 99.9% coulombic efficiency in a Ti||Zn asymmetric cell for 1,000 cycles; steady charge–discharge in a Zn||Zn symmetric cell for 6,000 cycles (6,000 h); and high energy densities (136 Wh kg−1 in a Zn||VOPO4 full battery with 88.7% retention for >6,000 cycles, 325 Wh kg−1 in a Zn||O2 full battery for >300 cycles and 218 Wh kg−1 in a Zn||MnO2 full battery with 88.5% retention for 1,000 cycles) using limited zinc. The SEI-forming electrolyte also allows the reversible operation of an anode-free pouch cell of Ti||ZnxVOPO4 at 100% depth of discharge for 100 cycles, thus establishing aqueous zinc batteries as viable cell systems for practical applications. A solid–electrolyte interphase that is permeable to Zn(ii) ions but waterproof is formed using an aqueous electrolyte composition. Cycling performances in an anode-free aqueous pouch cell show promise for intrinsically safe energy storage applications.

Arginine (Arg) metabolism is associated with many cellular and developmental processes in plants and proline, nitric oxide (NO) and polyamines (PAs) have a wide range of physiological functions in plants, including increased tolerance to environmental stresses. This study aimed to test the hypothesis that Arg spraying would stimulate the synthesis of proline, NO and PAs, reducing the oxidative damage caused by water deficit (WD) and increasing drought tolerance of sugarcane plants. Sugarcane plants were sprayed with water or Arg 1 mM, and subjected to WD by gradual addition of polyethylene glycol (PEG-8000) to the nutrient solution. As references, sugarcane plants were grown in nutrient solution without PEG-8000 and sprayed or not with Arg. Our data indicate that exogenous Arg supply improved leaf gas exchange during water deficit and enhanced the root antioxidative protection of sugarcane plants during the recovery period. Arg supply prevented the proline accumulation induced by water deficit and then the main pathway for proline synthesis is likely through glutamate instead of arginine. Although Arg is a substrate for NO and PAs production, supplying Arg had only slight effects in both NO and PAs levels. The spraying of amino acids capable of reducing the harmful effects of drought, such as Arg, can be an alternative to improve crop growth under field conditions.

The present study demonstrated that chitin-based nanofibers (CNFs) trigger the chitinase genes (PGIP1 and CaChi2), while elevating salicylic acid that can protect plants against pathogens. Cross-talk between this genetic induction and salicylic-acid-mediated immune response was also observed, which may arm a plant against multiple pathovars. Crab and mushroom based CNFs were synthesized by electrospinning and ball milling techniques. Plants (mung bean, Vigna radiata) (pepper, Capsicum annuum) were pre-inoculated with CNFs and treated with the pathogens Scrolotium rolfsii for pepper and Macrophomina phaseolina for mung bean and shrimp-based CNFs were used as a control. Treated plants had elevated levels of chitinase genes in response to CNFs at inoculation concentrations10 mg/mL both in soil and media, to protect them against the pathogenic fungal disease. After 24 h of exposure to the pathogens, qRT-PCR showed genes class II chitinase gene (CaChi2) and polygalacturonase inhibitor protein 1 (PGIP1) to be up-regulated in both root and shoot at 0.1 and 1 mg/mL of inoculation, respectively. The ball milled mushroom CNFs were sufficient to trigger the membrane based enzymes with less diameter (≥15 nm) to be most efficient versus others. In vitro analysis showed IC

CD28 is required for T cell activation as well as the generation of CD4+Foxp3+ Treg. It is unclear, however, how CD28 costimulation affects the development of CD8+ T cell suppressive function. Here, by use of Hepa1.6.gp33 in vitro killing assay and B16.gp33 tumor mouse model we demonstrate that CD28 engagement during TCR ligation prevents CD8+ T cells from becoming suppressive. Interestingly, our results showed that ectonucleotidase CD73 expression on CD8+ T cells is upregulated in the absence of CD28 costimulation. In both murine and human tumor-bearing hosts, CD73 is upregulated on CD28−CD8+ T cells that infiltrate the solid tumor. UPLC-MS/MS analysis revealed that CD8+ T cells activation without CD28 costimulation produces elevated levels of adenosine and that CD73 mediates its production. Adenosine receptor antagonists block CD73-mediated suppression. Our data support the notion that CD28 costimulation inhibits CD73 upregulation and thereby prevents CD8+ T cells from becoming suppressive. This study uncovers a previously unidentified role for CD28 costimulation in CD8+ T cell activation and suggests that the CD28 costimulatory pathway can be a potential target for cancer immunotherapy. Lai et al. report that the immunosuppressive molecule CD73 is negatively regulated by CD28 costimulation during CD8+ T cell activation. Their study implicates a lack of CD28 costimulation renders CD8+ T cells immunosuppressive and less able to eliminate solid tumors.

Collective cell migration is a fundamental biological process in which groups of cells move together in a coordinated manner, and it is essential for tissue development and wound repair. However, the underlying mechanisms that orchestrate directionality in collectively migrating cells remain poorly understood. In this study, we employed dynamically adhesive micropatterned substrates to investigate the role of adhesive cues in directing epithelial migration. Our findings demonstrate that epithelial cells collectively polarize in response to asymmetric patterns of extracellular matrix (ECM), and the degree of polarization depends on the degree of asymmetry and requires calcium-dependent cell-cell adhesion. When released from the micropatterns, epithelial cells collectively migrate according to the direction of pre-established polarity, and cohesive migration specifically requires E-cadherin-containing adherens junctions. Finally, disruption of the microtubule network blocks collective polarization and functionally inhibits directed migration. Together, these results indicate that adhesive cues from the ECM guide collective epithelial polarity and migration, and this response depends on adherens junctions and microtubules. STATEMENT OF SIGNIFICANCE: This study employs a dynamically adhesive micropatterning platform to investigate the role of adhesive cues in directing the polarity and directional migration of epithelial cells. The findings demonstrate how asymmetric tissue geometry influences the collective directionality in simple epithelia and that this response is mediated by adherens junctions and the microtubule network. This work provides new insight into fundamental cellular processes involved in wound healing and has important implications for biomaterial and scaffold design.

High thermal conductivity materials show promise for thermal mitigation and heat removal in devices. However, shrinking the length scales of these materials often leads to significant reductions in thermal conductivities, thus invalidating their applicability to functional devices. In this work, we report on high in-plane thermal conductivities of 3.05, 3.75, and 6 μm thick aluminum nitride (AlN) films measured via steady-state thermoreflectance. At room temperature, the AlN films possess an in-plane thermal conductivity of ∼260 ± 40 W m-1 K-1, one of the highest reported to date for any thin film material of equivalent thickness. At low temperatures, the in-plane thermal conductivities of the AlN films surpass even those of diamond thin films. Phonon-phonon scattering drives the in-plane thermal transport of these AlN thin films, leading to an increase in thermal conductivity as temperature decreases. This is opposite of what is observed in traditional high thermal conductivity thin films, where boundaries and defects that arise from film growth cause a thermal conductivity reduction with decreasing temperature. This study provides insight into the interplay among boundary, defect, and phonon-phonon scattering that drives the high in-plane thermal conductivity of the AlN thin films and demonstrates that these AlN films are promising materials for heat spreaders in electronic devices.

Phase separation of biological molecules, such as nucleic acids and proteins, has garnered widespread attention across many fields in recent years. For instance, liquid-liquid phase separation has been implicated not only in membraneless intracellular organization but also in many biochemical processes, including transcription, translation, and cellular signaling. Here, we present a historical background of biological phase separation and survey current work on nuclear organization and its connection to DNA phase separation from the perspective of DNA sequence, structure, and genomic context.

Delta-like ligand 3 (DLL3) is a therapeutic target for the treatment of small cell lung cancer, neuroendocrine prostate cancer, and isocitrate dehydrogenase mutant glioma. In the clinic, DLL3-targeted 89Zr-immunoPET has the potential to aid in the assessment of disease burden and facilitate the selection of patients suitable for therapies that target the antigen. The overwhelming majority of 89Zr-labeled radioimmunoconjugates are synthesized via the random conjugation of desferrioxamine (DFO) to lysine residues within the immunoglobulin. While this approach is admittedly facile, it can produce heterogeneous constructs with suboptimal in vitro and in vivo behavior. In an effort to circumvent these issues, we report the development and preclinical evaluation of site-specifically labeled radioimmunoconjugates for DLL3-targeted immunoPET. To this end, we modified a cysteine-engineered variant of the DLL3-targeting antibody SC16-MB1 with two thiol-reactive variants of DFO: one bearing a maleimide moiety (Mal-DFO) and the other containing a phenyloxadiazolyl methyl sulfone group (PODS-DFO). In an effort to obtain immunoconjugates with a DFO-to-antibody ratio (DAR) of 2, we explored both the reduction of the antibody with tris(2-carboxyethyl) phosphine (TCEP) as well as the use of a combination of glutathione and arginine as reducing and stabilizing agents, respectively. While exerting control over the DAR of the immunoconjugate proved cumbersome using TCEP, the use of glutathione and arginine enabled the selective reduction of the engineered cysteines and thus the formation of homogeneous immunoconjugates. A head-to-head comparison of the resulting 89Zr-radioimmunoconjugates in mice bearing DLL3-expressing H82 xenografts revealed no significant differences in tumoral uptake and showed comparable radioactivity concentrations in most healthy nontarget organs. However, 89Zr-DFOPODS-DAR2SC16-MB1 produced 30% lower uptake (3.3 ± 0.5 %ID/g) in the kidneys compared to 89Zr-DFOMal-DAR2SC16-MB1 (4.7 ± 0.5 %ID/g). In addition, H82-bearing mice injected with a 89Zr-labeled isotype-control radioimmunoconjugate synthesized using PODS exhibited ∼40% lower radioactivity in the kidneys compared to mice administered its maleimide-based counterpart. Taken together, these results demonstrate the improved in vivo performance of the PODS-based radioimmunoconjugate and suggest that a stable, well-defined DAR2 radiopharmaceutical may be suitable for the clinical immunoPET of DLL3-expressing cancers.

We greatly appreciate the financial support from the National Key Research & Development Project (2020YFB1506304, 2017YFE0129300), National Natural Science Foundation of China (52072135), and China Scholarship Council (201806160178). The oxygen electrode with a fast oxygen reduction reaction (ORR), oxygen evolution reaction (OER), and sufficient durability plays a pivotal role in reversible solid oxide cells (RSOCs). Here, we demonstrate a NdBa0.5Ca0.5Co1.5Fe0.5O5+δ@Gd0.1Ce0.9O2−δ (NBCCF@GDC) composite oxygen electrode via a one-pot method for exhibiting strong coherency, which result in boosting the electrochemical performance of RSOCs. The NBCCF@GDC electrode yields a very low polarization resistance (0.106 Ω-cm2 at 800 °C), high electrolysis current density (1.45 A cm–2 with 70 vol % absolute humidity at 1.3 V), and high power density (∼1.3 W cm–2 at 800 °C) and shows excellent reversibility and stability. Notably, strong coherency in these NBCCF@GDC composite materials was successfully revealed by HT-XRD, XPS, STEM, and EELS. The phase contiguity and interfacial coherence between NBCCF and GDC increase the Co oxidation state and the number of active sites, which enhanced the electrocatalytic activity for perovskites. Overall, this work demonstrates a highly desirable strategy for the production of functionalized electrodes for next-generation reversible solid oxide cells. Postprint

When studying a new electrochemical couple, numerous variables affect how it is evaluated, how successful it is judged to be, and system properties. In the energy storage field, many researchers evaluate materials using performance metrics that might include capacity, rate capability, or cell impedance values to assess various cell chemistries. Behind these numbers are underlying attributes such as particle morphology, tap density, particle size, surface functionality, and even growth facet that are influenced by the method synthesis. These characteristics can carry through to the final product as the material is transformed into an electrode, high-surface-area catalyst support, or a stand-alone sintered ceramic. In this article, the focus will be on, at a high level, how the synthesis procedure chosen and the variables that come with that choice influence the observed properties of the final product.

To compare the prevalence of intravitreal silicone oil microdroplets detected by slit-lamp biomicroscopy in eyes with 6 or more injections of the same anti-vascular endothelial growth factor (VEGF) drug.Prospective, cross-sectional case series.A total of 260 consecutive eyes receiving 1 of 3 intravitreal anti-VEGF drugs for choroidal neovascularization, diabetic macular edema, or venous occlusive disease. The control group included 147 fellow eyes with no prior intravitreal injections.The anterior and mid-vitreous were carefully examined using 12× to 16× magnification through dilated pupils with ocular saccades before an injection. Silicone oil microdroplets were graded on a scale from 0 to 4+ based on the number and size of droplets.Presence and severity of silicone oil microdroplets in the vitreous.Silicone oil microdroplets were observed in 78.3% of eyes receiving bevacizumab in Becton Dickinson (BD, Franklin Lakes, NJ) 0.3-mL polypropylene syringes, 14.4% of eyes receiving ranibizumab in 1.0-mL BD polypropylene syringes or more recently glass prefilled syringes, 48.5% of eyes receiving aflibercept in 1.0-mL BD polycarbonate syringes, and 0% of eyes in controls. The differences among the 4 groups were statistically significant at P0.001. The severity of silicone oil microdroplets was significantly greater in eyes using BD 0.3-mL polypropylene syringes than BD 1.0-mL polypropylene syringes, BD 1.0-mL polycarbonate syringes, or controls (P0.001). The severity of silicone oil microdroplets in eyes using BD 1.0-mL polycarbonate syringes was significantly greater than BD 1.0-mL polypropylene syringes (P = 0.012) and controls (P0.001). There was no significant difference between silicone oil microdroplet severity between BD 1.0-mL polypropylene syringes and controls (P = 1.0).The BD 0.3-mL polypropylene syringes with repackaged bevacizumab and the BD 1.0-mL polycarbonate syringes with aflibercept cause a higher likelihood of silicone oil microdroplets. Intravitreal injections in eyes receiving multiple regular anti-VEGF injections should be supplied in silicone-free syringes.

The authors would like to thank the Petroleum Technology Development Fund (Nigeria) for funding this research and University of St Andrews (Scotland, UK) for the opportunity to carry out the research. Environmental problems associated with the use of fossil fuels and increase in energy demands due to rise in population and rapid industrialisation, are the driving forces for energy. Catalytic conversion of biomass to renewable energies is among the promising approaches to materialize the above. This requires development of robust catalysts to suppress deactivation due to carbon deposition and agglomeration. In this work, surface properties and chemistry such as exsolution of B-site metal catalyst nanoparticles, particle size and distribution, as well as catalyst-support interactions were tailored through the use of alkaline dopants to enhance catalytic behaviour in valorisation of glycerol. The incorporation of alkaline metals into the lattice of an A-site deficient perovskite modified the surface basic properties and morphology with a consequent robust catalyst-support interaction. This resulted in promising catalytic behaviour of the materials where hydrogen selectivity of over 30% and CO selectivity of over 60% were observed. The catalyst ability to reduce fouling of the catalyst surface as a result of carbon deposition during operation was also profound due to the robust catalyst-support interaction occurring at the exsolved nanoparticles due to their socketing and the synergy between the dopant metals in the alloy in perovskite catalyst systems. In particular, one of the designed systems, La0.4Sr0.2Ca0.3Ni0.1Ti0.9O3±δ, displayed almost 100% resistance to carbon deposition. Therefore, lattice rearrangement using exsolution and choice of suitable dopant could be tailored to improve catalytic performance. Publisher PDF

Two proteins within the β-grasp superfamily, the B1-domain of protein G and the small archaeal modifier protein 1, were investigated to elucidate the key determinants of structural stability at the level of individual interactions. These symmetrical proteins both contain two β-hairpins which form a sheet flanked by a central α-helix. They were subjected to high temperature molecular dynamics simulations and the detailed behavior of each long-range interaction was characterized. The results revealed that in GB1 the most stable region was the C-terminal hairpin and in SAMP1 it was the opposite, the N-terminal hairpin. Experimental results for GB1 support this finding. In conclusion, it appears that the difference in the location and number of hydrophobic interactions dictate the differential stability which is accommodated due to structural symmetry of the β-grasp fold. Thus, the hairpins are interchangeable and in nature this lends itself to adaptability and flexibility.

The rapid onset of the global COVID-19 pandemic has led to multiple challenges for accurately diagnosing the infection. One of the main bottlenecks for COVID-19 detection is reagent and material shortages for sample collection, preservation, and purification prior to testing. Currently, most authorized diagnostic tests require RNA extraction from patient samples and detection by reverse transcription polymerase chain reaction (RT-PCR). However, RNA purification is expensive, time consuming, and requires technical expertise to perform. Additionally, there have been reported shortages of the RNA purification kits needed for most tests. With these challenges in mind, we report on extraction-free amplification of SARS-CoV-2 RNA directly from patient samples. In addition, we have developed a multiplex RT-PCR using the CDC singleplex targets. This multiplex has a limit of detection of 2 copies/μL. We have demonstrated these improvements to the current diagnostic workflow, which reduce complexity and cost, minimize reagent usage, expedite time to results, and increase testing capacity.

Background: Surrogate monoclonal antibodies (mAbs) used in preclinical in vivo studies can be challenging to quantify due to lack of suitable immunoaffinity reagents or unavailability of the mAb protein sequence. Generic immunoaffinity reagents were evaluated to develop sensitive LC–MS/MS assays. Peptides of unknown sequence can be used for selective LC–MS quantification. Results: anti-mouse IgG1 was found to be an effective immunoaffinity reagent, enabling quantification of mouse IgG1 mAbs in mouse serum. Selective peptides of unknown sequence were applied for multiplex LC–MS quantification of two rat mAbs co-dosed in mouse. Conclusion: Generic anti-mouse IgG subtype-specific antibodies can be used to improve assay sensitivity and peptides of unknown sequence can be used to quantify surrogate mAbs when the mAb protein sequence in unavailable.

Background Cyclical, high-dose testosterone administration, termed bipolar androgen therapy (BAT), can induce clinical responses and restore sensitivity to androgen signalling inhibition in patients with previously treated castration-resistant prostate cancer (PCa) (CRPC). This trial evaluated whether BAT is a safe and effective first-line hormonal therapy for patients with CRPC. Patients and Methods In cohort C of this single-centre, open-label, phase II, multi-cohort trial (RE-sensitizing with Supraphysiologic Testosterone to Overcome REsistance study), 29 patients with CRPC received first-line hormonal therapy with 400 mg of testosterone cypionate intramuscularly every 28 days concurrent with a luteinising hormone-releasing hormone agonist/antagonist. The primary end-point of the study was the PSA50 response rate to BAT treatment. Results After treatment with BAT, four of 29 patients (14%; 95% confidence interval [CI]: 4–32%) experienced a PSA50 response. The median radiographic progression-free survival to BAT was 8.5 months (95% CI: 6.9–15.1) for patients with metastatic CRPC. After progression on BAT, 17 of 18 patients (94%; 95% CI: 73–100%) achieved a PSA50 response and 15 of 18 patients (83%; 95% CI: 59–96) achieved a PSA90 response on abiraterone or enzalutamide. Twelve of 15 patients (80%; 95% CI: 52–96) with metastatic CRPC remain on abiraterone or enzalutamide with a median duration of follow-up of 11.2 months. Conclusion As first-line hormonal treatment for CRPC, BAT was well tolerated and resulted in prolonged disease stabilisation. After progression on BAT, patients had favourable responses to second-generation androgen receptor–targeted therapy. Trial Registration ClinicalTrials.gov NCT02090114

In recent years, there has been an increasing interest in designing delivery systems to enhance the efficacy of RNA-based therapeutics. Here, we have synthesized copolymers comprised of dimethylaminoethyl methacrylate (DMAEMA) or diethylaminoethyl methacrylate (DEAEMA) copolymerized with alkyl methacrylate monomers ranging from 2 to 12 carbons, and developed a high throughput workflow for rapid investigation of their applicability for mRNA delivery. The structure activity relationship revealed that the mRNA encapsulation efficiency is improved by increasing the cationic density and use of shorter alkyl side chains (2-6 carbons). Minimal cytotoxicity was observed when using DEAEMA

A critical new research direction in solid oxide cells (SOCs) relates to balancing power grid or integrating energy interconnection with heat-electricity-gas simply by operating under different modes including solid oxide fuel cell and electrolyser cell. The rational design of robust and high-performance materials for SOCs is urgent for high conversion/energy efficiencies. Iron is highly earth abundant and offers suitably flexible redox chemistry to address such operational modes. Iron-based oxide materials for SOCs are widely investigated because of the low cost and, but more importantly, the appropriate valence stability of the Fe-O bond, which affords good redox activity across a wide range of electrode functions. This review describes the progress in iron-based materials for SOCs, especially the recent applications in electrode materials or catalysts. The stable structure of the ferrite oxide allows and important platform for improved performance via substitution of Fe to deliver high-performance fuel electrodes of an SOC with H2/H2O or carbonaceous fuel/feedstock. Furthermore, we discuss nano-sized Fe0 metal or alloys on oxide electrode via infiltration and in situ exsolution aiming to fabricate high active electrocatalysts. The advances of ferrite oxide-based oxygen electrode are also discussed in terms of thermal expansion, structural stability and electrocatalysis before the development of symmetrical and reversible SOCs based on ferrite oxides are classified and summarized. Thereby, the challenges and future prospects are discussed.

Objective Normalized creatinine generation rate (CGRn) can be computed for a variety of dialysis schedules using a recently described kinetic modeling program. However, the availability of estimating equations might facilitate broader study of this metric. We developed equations to estimate CGRn based on modeling and then tested them against modeled CGRn values in the Frequent Hemodialysis Network Nocturnal Trial baseline (3/week) dataset. Design and Methods We used a "what-if" derivation of a previously published variable volume 2-pool creatinine kinetic model to generate predicted predialysis values of serum creatinine that would result from creatinine generation rates of 250–2000 mg/day in patients with creatinine distribution volumes of 20 to 50 L, dialyzed from 60 to 480 min per treatment three times a week. Then, in patients with residual kidney function, we calculated an "anuric expected predialysis serum creatinine value" before applying the same equations. We then compared estimated CGRn values as predicted by this approach with modeled values in patient data from the Frequent Hemodialysis Network Nocturnal Trial. Results The estimating equations for CGRn yielded results similar to those obtained with formal modeling, in both anuric patients and those with residual kidney function, with mean percent error of 0.845 ± 6.15 (SD) in anuric patients, and ‒0.29 ± 4.90 in patients with a mean creatinine clearance of 5.44 ± 4.82 mL/min, with R-squared values of 0.96 in both anuric patients and those with residual renal clearance of creatinine. Conclusions In patients dialyzed 3/week, CGRn can be estimated using prediction equations. Use of these equations may facilitate broader investigation of CGRn as a measure of nutritional status and outcome.

HighlightsConcern over nutrient runoff from agriculture has prompted research to limit phosphorus (P) mobility.Mine drainage residuals (MDRs) can reduce phosphorus solubility in soils, sediments and liquid manure slurries.MDR amendment resulted in lower dissolved phosphorus in runoff from sites treated with dairy manure slurry, but not with swine manure slurry.This study underscores the value of testing under field conditions before making recommendations.Abstract. Concern over nutrient runoff from agriculture has prompted considerable research on amendments to limit phosphorus (P) solubility of manure slurries and P mobility following land application of the slurry. The concept of solving one industry’s problem with another industry’s problem is attractive, but successful examples are uncommon. Mine drainage residual (MDR), generated from the process of neutralizing acid discharge from coal mines, has been shown to reduce soluble P in soils, sediments and manure slurries. We therefore sought to test whether amending manure slurries with MDR was effective at reducing P in runoff once that slurry was applied to agricultural soils. A series of simulated rainfall experiments revealed that amending dairy manure slurry with MDR resulted in significantly less flow weighted dissolved P concentrations and loads in runoff. However, the same effect was not observed with runoff from soils amended with swine manure slurry, despite a greater reduction of water extractable P in swine manure slurry with MDR addition than in dairy manure slurry. This study underscores the value of testing amendments under field conditions before making manure management recommendations. Keywords: Manure, Phosphorus, Mine drainage residual, Engineered treatment, Simulated rainfall.

Organic aerosols, a major constituent of fine particulate mass in megacities, can be directly emitted or formed from secondary processing of biogenic and anthropogenic volatile organic compound emissions. The complexity of volatile organic compound emission sources, speciation and oxidation pathways leads to uncertainties in the key sources and chemistry leading to formation of organic aerosol in urban areas. Historically, online measurements of organic aerosol composition have been unable to resolve specific markers of volatile organic compound oxidation, while offline analysis of markers focus on a small proportion of organic aerosol and lack the time resolution to carry out detailed statistical analysis required to study the dynamic changes in aerosol sources and chemistry. Here we use data collected as part of the joint UK-China Air Pollution and Human Health (APHH-Beijing) collaboration during a field campaign in urban Beijing in the summer of 2017 alongside laboratory measurements of secondary organic aerosol from oxidation of key aromatic precursors (1,3,5-Trimethyl benzene, 1,2,4-Trimethyl benzene, propyl benzene, isopropyl benzene and 1-methyl naphthalene) to study the anthropogenic and biogenic contributions to organic aerosol. For the first time in Beijing, this study applies positive matrix factorisation to online measurements of organic aerosol composition from a time-of-flight iodide chemical ionisation mass spectrometer fitted with a filter inlet for gases and aerosols (FIGAERO-ToF-I-CIMS). This approach identifies the real-Time variations in sources and oxidation processes influencing aerosol composition at a near-molecular level. We identify eight factors with distinct temporal variability, highlighting episodic differences in OA composition attributed to regional influences and in situ formation. These have average carbon numbers ranging from C5-C9 and can be associated with oxidation of anthropogenic aromatic hydrocarbons alongside biogenic emissions of isoprene, α-pinene and sesquiterpenes. This journal is