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Simultaneous poisoning of sulfur + phosphorus on a platinum-based diesel oxidation catalyst was studied to gain a deeper understanding on the catalyst deactivation. Compared to a single poisoning (sulfur or phosphorus), the simultaneous poisoning had a severe effect on the catalyst activation: light-off temperature for 90% conversion of propene was not reached, this of carbon monoxide was much higher than expected, and the maximum conversion of nitrogen monoxide collapsed. With very comprehensive structural characterization by various methods (e.g. STEM-EDS, XPS, DRIFTS) used, we achieved to conclude an explanation for this. In the case of the S + P-poisoning of the catalyst, formed aluminum phosphate was found to block adsorption sites for sulfur species on alumina and sulfur adsorbs mainly on cerium oxides. In addition, sulfur species remain with and in the vicinity of the platinum particles blocking active sites. publishedVersion

20 p.-8 fig., Low-density lipoprotein (LDL)-cholesterol delivery from late endosomes to the plasma membrane regulates focal adhesion dynamics and cell migration, but the mechanisms controlling it are poorly characterized. Here, we employed auxin-inducible rapid degradation of oxysterol-binding protein-related protein 2 (ORP2/OSBPL2) to show that endogenous ORP2 mediates the transfer of LDL-derived cholesterol from late endosomes to focal adhesion kinase (FAK)-/integrin-positive recycling endosomes in human cells. In vitro, cholesterol enhances membrane association of FAK to PI(4,5)P2-containing lipid bilayers. In cells, ORP2 stimulates FAK activation and PI(4,5)P2 generation in endomembranes, enhancing cell adhesion. Moreover, ORP2 increases PI(4,5)P2 in NPC1-containing late endosomes in a FAK-dependent manner, controlling their tubulovesicular trafficking. Together, these results provide evidence that ORP2 controls FAK activation and LDL-cholesterol plasma membrane delivery by promoting bidirectional cholesterol/PI(4,5)P2 exchange between late and recycling endosomes., This study was supported by the Academy of Finland (grants 307415 and 324929 to E.I., 322647 to V.M.O.), Sigrid Juselius Foundation (E.I., V.M.O., L. A.-S.), Fondation Leducq (grant 19CVD04) (E.I.), Jane and Aatos Erkko Foundation (E.I.), Magnus Ehrnrooth Foundation (K.T., V.M.O.), and the Finnish Foundation for Cardiovascular Research (V.M.O.). L.-A.S. acknowledges support from Helsinki Institute of Life Science (HiLIFE, start-up grant) and D.L. from the Ministry of Science, Innovation and Universities for the Spanish State Research Agency Retos Grant RTI2018-099318-B-I00, cofounded by the European Regional Development Fund (FEDER).

Oncolytic adenoviruses are promising cancer therapeutic agents. Clinical data have shown adenoviruses’ ability to transduce tumors after systemic delivery in human cancer patients, despite antibodies. In the present work, we have focused on the interaction of a chimeric adenovirus Ad5/3 with human lymphocytes and human erythrocytes. Ad5/3 binding with human lymphocytes and erythrocytes was observed to occur in a reversible manner, which allowed viral transduction of tumors, and oncolytic potency of Ad5/3 in vitro and in vivo, with or without neutralizing antibodies. Immunodeficient mice bearing xenograft tumors showed enhanced tumor transduction following systemic administration, when Ad5/3 virus was bound to lymphocytes or erythrocytes (P

Composite fibers based on a polylactide (PLA) matrix with the addition of dispersed spherical particles, namely chitin nanoparticles modified with polyethylene glygol (PEG) and silver nanoparticles modified with polyvinylpyrrolidone, have been obtained using a twin-screw microextruder. The resulting fibers were subjected to high-temperature orientational drawing four and six times. The features of the mechanical properties and supramolecular structure of oriented composite fibers have been studied. The similar character of the curves of the tensile diagrams of PLA fibers and composite fibers based on it is shown. A gradual increase in the strength of composite fibers elongated by a factor of four with an increase in the concentration of spherical particles has been established. In the case of fiber drawing by a factor of six, the maximum strength is observed by an average of 20% at a concentration of silver nanoparticles of 1 wt %, and chitin nanoparticles 5 wt %. The values of deformation and elastic modulus of composite fibers remain close to those of unfilled PLA fibers. It has been found that composite fibers have a highly oriented fibrillar structure. Good adhesion of the PLA matrix to the surface of PEG-modified chitin particles has been shown

aintenance therapy containing methotrexate and 6-mercapto-purine is essential to cure acute lymphoblastic leukemia (ALL). Cytotoxicity is elicited by incorporation of thioguanine nucleotides into DNA (DNA-TG), and higher leukocyte DNA-TG is associated with increased relapse-free survival. As 6-thioguanine provides 6-fold higher cytosolic levels of thioguanine nucleotides than does 6-mercaptopurine, we added low-dose 6-thioguanine to methotrexate/6mercaptopurine maintenance therapy to explore if this combination results in significantly higher DNA-TG. The target population of the “Thiopurine Enhanced ALL Maintenance therapy” (TEAM) study was 30 patients with non-high-risk ALL, aged 1-45 years on methotrexate/6-mercaptopurine maintenance therapy receiving no other systemic chemotherapy. Incremental doses of 6-thioguanine were added to methotrexate/6-mercaptopurine maintenance therapy (starting 6-thioguanine dose: 2.5 mg/m2/day, maximum: 12.5 mg/m2/day). The primary endpoint was DNA-TG increments. Thirty-four patients were included, and 30 patients completed maintenance therapy according to the TEAM strategy. Of these 30 patients, 26 (87%) tolerated 10.0-12.5 mg/m2/day as the maximum 6-thioguanine dose. TEAM resulted in significantly higher DNA-TG levels compared to those in both TEAM patients before their inclusion in TEAM (on average 251 fmol/mg DNA higher [95% confidence interval: 160-341; P

The intracellular transport of cholesterol is subject to tight regulation. The structure of the lysosomal integral membrane protein type 2 (LIMP-2, also known as SCARB2) reveals a large cavity that traverses the molecule and resembles the cavity in SR-B1 that mediates lipid transfer. The detection of cholesterol within the LIMP-2 structure and the formation of cholesterol−like inclusions in LIMP-2 knockout mice suggested the possibility that LIMP2 transports cholesterol in lysosomes. We present results of molecular modeling, crosslinking studies, microscale thermophoresis and cell-based assays that support a role of LIMP-2 in cholesterol transport. We show that the cavity in the luminal domain of LIMP-2 can bind and deliver exogenous cholesterol to the lysosomal membrane and later to lipid droplets. Depletion of LIMP-2 alters SREBP-2-mediated cholesterol regulation, as well as LDL-receptor levels. Our data indicate that LIMP-2 operates in parallel with Niemann Pick (NPC)-proteins, mediating a slower mode of lysosomal cholesterol export., Cholesterol transport is tightly regulated in the cell and in lysosomes is regulated by NPC1/2. Here, Heybrock et al. use molecular modeling, knockout mice and cell based studies to show that LIMP-2 also mediates lysosomal cholesterol transport.

Metal elements, ubiquitous in the environment, can cause negative effects in long-lived organisms even after low but prolonged exposure. Insectivorous bats living near metal emission sources can be vulnerable to such contaminants. Although it is known that bats can bioaccumulate metals, little information exists on the effects of metal elements on their physiological status. For example, oxidative status markers are known to vary after detoxification processes and immune reactions. Here, for two consecutive summers, we sampled individuals from a natural population of the insectivorous bat, Myotis daubentonii, inhabiting a site close to a metal emission source. We quantified metals and metalloids (As, Ca, Cd, Co, Cu, Mn, Ni, Pb, Se, Zn) from individual fecal pellets. We measured enzymatic antioxidants (GP, CAT, SOD), total glutathione (tGSH) and ratio between reduced and oxidized glutathione (GSH:GSSG) from their red blood cells together with biometrics, hematocrit and parasite prevalence. In general, metal concentrations in feces of M. daubentonii reflected the exposure to ambient contamination. This was especially evident in the higher concentrations of Cd, Co, Cu and Ni close to a smelter compared to a site with less contaminant exposure. Annual differences were also observed for most elements quantified. Sex-specific differences were observed for calcium and zinc excretion. SOD and CAT enzymatic activities were associated with metal levels (principal components of six metal elements), suggesting early signs of chronic stress in bats. The study also shows promise for the use of non-invasive sampling to assess the metal exposure on an individual basis and metal contamination in the environment.

Reduced graphene oxide (rGO) integrated nano-fibrillated cellulose (NFC) grafted poly(ionic liquid) (PIL) composite paper with enhanced hydrophobicity, ductility and toughness was fabricated by vacuum filtration of an aqueous dispersion of rGO/NFC-graft-PIL (NFC-g-PIL). The NFC-g-PIL was synthesized by growing PIL chains from the NFC surface using free radical polymerization of a pyridinium acrylate-based ionic liquid monomer. publishedVersion

Publisher Copyright: © 2021 Lippincott Williams and Wilkins. All rights reserved. Asparaginase-associated pancreatitis (AAP) frequently affects children treated for acute lymphoblastic leukemia (ALL) causing severe acute and persisting complications. Known risk factors such as asparaginase dosing, older age and single nucleotide polymorphisms (SNPs) have insufficient odds ratios to allow personalized asparaginase therapy. In this study, we explored machine learning strategies for prediction of individual AAP risk. We integrated information on age, sex, and SNPs based on Illumina Omni2.5exome-8 arrays of patients with childhood ALL (N=1564, 244 with AAP aged 1.0 to 17.9 y) from 10 international ALL consortia into machine learning models including regression, random forest, AdaBoost and artificial neural networks. A model with only age and sex had area under the receiver operating characteristic curve (ROC-AUC) of 0.62. Inclusion of 6 pancreatitis candidate gene SNPs or 4 validated pancreatitis SNPs boosted ROC-AUC somewhat (0.67) while 30 SNPs, identified through our AAP genome-wide association study cohort, boosted performance (0.80). Most predictive features included rs10273639 (PRSS1-PRSS2), rs10436957 (CTRC), rs13228878 (PRSS1/PRSS2), rs1505495 (GALNTL6), rs4655107 (EPHB2) and age (1 to 7 y). Second AAP following asparaginase re-exposure was predicted with ROC-AUC: 0.65. The machine learning models assist individual-level risk assessment of AAP for future prevention trials, and may legitimize asparaginase re-exposure when AAP risk is predicted to be low.

Aims: The aim was to examine whether the contribution of physical work exposures to the risk of sickness absence (SA) is different between those with and without common mental disorders (CMD). Methods: We used questionnaire data on four work exposures and CMD from 6159 participants of the Helsinki Health Study cohort with 12,458 observations from three surveys (2000–2002, 2007 and 2012). We formed combination exposures for the work exposures (hazardous exposures, physical workload, computer and shift work) with CMD. Associations with SA of different length were examined with negative binomial regression models. Results: We observed stronger associations for CMD with SA than for the individual work exposures. The strength of the associations for hazardous exposures and physical workload increased with length of SA, especially when the participant also had CMD. The strongest associations for the combined exposures were observed for SA ⩾15 days, the rate ratios being 2.63 (95% CI 2.27–3.05) among those with hazardous exposure and CMD, and 3.37 (95% CI 2.93−3.88) among those with heavy physical workload and CMD. Conclusions: Employees with hazardous exposures or physical workload combined with CMD were at the highest risk of SA compared with those without these exposures or with only one exposure.

Despite substantial progress being made relating to 2D-nanofiller-based composite nanolaminates, the fabrication of composite nanolaminates with enhanced ductility and toughness is still challenging. In this study, layered structure graphene oxide (GO)/gelatin powder (GP) composites nanolaminates with enhanced ductility and toughness have been achieved by a simple vacuum filtration of aqueous dispersion of GO/GP composite solution containing 5 wt% of GO. The composite film containing 5 wt% GO shows outstanding improvement of 200% in the stress at break value, with simultaneous enhancement of 52% of the strain at break value compared to GP film. A significant improvement in toughness from 2.2 MJ m-3 to 9.5 MJ m-3 is observed in the composite film containing 5 wt% GO. These significant enhancements of the mechanical properties of the composite film are obtained via the formation of an intercalated nanolaminate structure, H-bonding interactions, and the tailoring of the crystal structure of GP in the composite film, as proved via field-emission scanning electron microscopy, Fourier-transform infrared spectroscopy, and wide-angle X-ray diffraction studies. The growing of fibroblast cells on the composite films signifies that they are not cytotoxic. These GO/GP composites with significant mechanical properties and biocompatibility are very promising for various biomedical applications. This journal is publishedVersion

The fitness and recruitment of fish stocks can be markedly affected by environmental disturbances including global warming, eutrophication and contamination. Understanding the effects of environmental stressors on salmon physiology during marine residence is of a global concern as marine survival has decreased. We present a unique combination of physiological responses - antioxidant defence and oxidative damage biomarkers, stable isotopes and contaminant exposure biomarkers - measured from adult Atlantic salmon (Salmo salar) collected at the Baltic Sea and studied in relation to environmental variables and fitness estimates. The results demonstrate that feeding populations of salmon display marked temporal and spatial variation in oxidative status. Better oxidative status of salmon was characterized by a higher amount of reduced glutathione (GSH) and decreased lipid peroxidation (LPX), when the weight-at-age of 3–4-year old sprats was higher and contaminant exposure biomarker (EROD) was lower. Summer season conditions, which included cooler sea surface temperature (SST), higher bottom O2 and less cyanobacteria also indicated conditions for better oxidative status. Summer SST was additionally shown to affected glutathione metabolism enzyme activities. Oxidative status was associated with stable isotopes δ13C and δ15N indicating indirect effect of abiotic conditions and lower levels of the food web. Differences in condition factor and growth were associated with oxidative status in one and two sea winter salmon, respectively. Wild salmon survival was higher in years when they had higher GSH and catalase activity and lower LPX. Enhanced glutathione metabolism and increased protein carbonyls were associated with higher occurrence of yolk-sac fry mortality (M74). Our results show that oxidative status can provide information on exposure to complex combinations of environmental conditions and stressors in the wild and provide a link of physiological function to individual and population level fitness effects. peerReviewed

The occurrence of triclosan (TCS) in the eggs of wild avian species is an emerging concern. We previously evaluated the effects of in ovo exposure to TCS on the liver transcriptome of chicken embryos and proposed adverse outcome pathways (AOPs). However, the key molecular events identified to be affected need to be verified at the protein level. Herein, we investigated the changes in the spectrum of hepatic proteins in TCS-treated chicken embryos by proteomic analysis to validate the key signaling pathways involved in the AOPs. We identified and quantified 894 unique proteins using matrix-assisted laser desorption/ionization time-of-flight/time-of-flight tandem mass spectrometry. In the 0.1 (low dose), 1 (median dose), and 10 μg triclosan/g egg (high dose) groups, TCS caused significant changes in the levels of 195, 233, and 233 proteins in males and 237, 188, and 156 proteins in females, respectively (fold changes1.3 or0.7). TCS exposure modulated the expression of proteins, predominantly involved in signaling pathways of lipid and energy metabolism in both genders. Among the proteins associated with TCS metabolism in the liver, phase I (e.g., CYP2C23a) and phase II (e.g., UGT1A1) enzymes mediated by chicken xenobiotic receptor, were only induced in males. In consonance with the malondialdehyde levels, which were increased upon TCS exposure in females in a dose-dependent manner, a battery of antioxidant enzymes, notably SOD2, GST, GSTz1, and PRDX1, was decreased and SOD1 and GSTK1 were increased in the embryos. Taken together, this proteome analysis complements the transcriptome profiling reported in our previous study and authenticates the AOPs proposed for chicken embryos in ovo exposed to TCS.

The immobilized enzyme lipase acts as an efficient, selective and durable catalyst in the direct transformation of unsaturated carboxylic acids to epoxides, which are used as chemical intermediates and bio-lubricants. Experimental data obtained from the epoxidation of a model molecule, oleic acid in a laboratory-scale isothermal batch reactor were critically evaluated and mathematically modelled in the most precise way. Several rival surface reaction mechanisms were proposed and rate equations based on these mechanisms were derived. The rate equations were implemented in a multiphase model for the laboratory-scale batch reactor and the kinetic and adsorption parameters included in the rate equations were estimated with non-linear regression analysis. Based on the parameter estimation statistics and chemical knowledge, the most plausible kinetic models for the chemo-enzymatic epoxidation of oleic acid on the immobilized lipase catalyst were selected. The best kinetic models gave a good reproduction of the experimental data. The models can be used to predict the performance of enzymatic epoxidation of unsaturated fatty acids.

Layered structure graphene oxide (GO)/methylcellulose (MC) composite films with excellent mechanical and gas barrier properties were achieved for the first time by a simple solvent evaporation assisted assembly of GO/MC composite solutions. 1, 3 and 5 wt% of GO composites with respect to MC were fabricated and denoted as GO1, GO3 and GO5, respectively. The interactions of the hydrophobic associated domains of MC (formed during solvent evaporation) with the π cloud of GO (hydrophobic interaction) and the oxygen-containing groups of MC with the oxygen-containing functional groups of GO (H-bonding interaction) govern the formation of self-assembled thick MC coated layered structure GO/MC composites. The obtained composite films were highly flexible and characterized by field emission scanning electron microscopy (FESEM), Fourier transform infrared spectroscopy (FTIR), wide angle X-ray scattering (WAXS), Simultaneous Thermal Analysis (STA), and mechanical and oxygen gas permeation studies. The FESEM analysis shows the formation of a layered structure as well as good dispersion of thick MC coated GO sheets in the MC matrix. WAXS studies also illustrate that the GO sheets remain in a highly exfoliated state in the MC matrix. The FTIR results indicate the presence of interfacial interaction between the GO and MC molecules. The GO/MC composite films exhibit a significant enhancement of mechanical properties compared to pure MC films. The stress at break and Young's modulus of GO5 were determined to be 104 ± 3 MPa and 3.8 ± 0.2, respectively. In addition to the mechanical properties, the oxygen gas barrier properties of the composites were also enhanced significantly. The GO1 film shows a 98% decrease of oxygen permeation with respect to pure MC films.

The aim of tissue engineering is the regeneration of damaged tissue or the production of representative tissue organoids in vitro. To achieve this, one approach is to use hydrogels, water-swollen hydrophilic and crosslinked polymer networks, that can accommodate encapsulation of living cells and help the regeneration process. Even though mechanically biomimicking target tissue is important for a favorable cell response, the mechanical characterisation of tissues or hydrogels is not yet a fully defined process with various possible models and methods existing. In this paper, for the first time, a specific procedure and model has been suggested for the discussion of the nonlinear stress-strain relationship in large deformations of hydrogels. Moreover, this approach has comprehensively characterised the compressive material performance of hydrogels in a theoretical framework. To present the performance and utility of the introduced procedure, it is used with two different compositions of bioamine crosslinked gellan gum hydrogel. In addition, a three-dimensional digital image correlation technique has been utilized together with compression testing to measure the actual force and deformation in unconfined compression. The material model parameters were obtained to represent nonlinear stress-strain behaviour and the viscoelastic response (relaxation) of gellan gum hydrogel in compression.

The isolation and physical material properties of suberin fatty acids (SFAs) were investigated with special reference to their potential applications as novel pharmaceutical excipients. SFAs were isolated from outer birch bark (OBB) with a new extractive hydrolysis method. The present simplified isolation process resulted in a moderate batch yield and chemical purity of SFAs, but further development is needed for establishing batch-to-batch variation. Cryogenic milling was the method of choice for the particle size reduction of SFAs powder. The cryogenically milled SFAs powder exhibited a semicrystalline structure with apparent microcrystalline domains within an amorphous fatty acids matrix. The thermogravimetric analysis (TGA) of SFAs samples showed a good thermal stability up to 200 °C, followed by a progressive weight loss, reaching a plateau at about 95% volatilization at about 470 °C. The binary blends of SFAs and microcrystalline cellulose (MCC; Avicel PH 101) in a ratio of 25:75 (w/w) displayed good powder flow and tablet compression properties. The corresponding theophylline-containing tablets showed sustained or prolonged-release characteristics. The physicochemical and bulk powder properties of SFAs isolated from OBB are auspicious in terms of potential pharmaceutical excipient applications.

Summary 3D printers are currently widely available and very popular among the general public. However, the use of these devices may pose health risks to users, attributable to air-quality issues arising from gaseous and particulate emissions in particular. We characterized emissions from a low-end 3D printer based on material extrusion, using the most common polymers: acrylonitrile-butadiene-styrene (ABS) and polylactic acid (PLA). Measurements were carried out in an emission chamber and a conventional room. Particle emission rates were obtained by direct measurement and modeling, whereas the influence of extrusion temperature was also evaluated. ABS was the material with the highest aerosol emission rate. The nanoparticle emission ranged from 3.7·108 to 1.4·109 particles per second (# s−1) in chamber measurements and from 2.0·109 to 4.0·109 # s−1in room measurements, when the recommended extruder temperature was used. Printing with PLA emitted nanoparticles at the rate of 1.0·107 # s−1 inside the chamber and negligible emissions in room experiments. Emission rates were observed to depend strongly on extruder temperature. The particles’ mean size ranged from 7.8 to 10.5 nanometers (nm). We also detected a significant emission rate of particles of 1 to 3 nm in size during all printing events. The amounts of volatile organic and other gaseous compounds were only traceable and are not expected to pose health risks. Our study suggests that measures preventing human exposure to high nanoparticle concentrations should be adopted when using low-end 3D printers.

Despite some promising results, the majority of patients do not benefit from T cell therapies, as tumors prevent T cells from entering the tumor, shut down their activity, or downregulate key antigens. Due to their nature and mechanism of action, oncolytic viruses have features that can help overcome many of the barriers currently facing T cell therapies of solid tumors. This study aims to understand how four different oncolytic viruses (adenovirus, vaccinia virus, herpes simplex virus, and reovirus) perform in that task. For that purpose, an immunocompetent in vivo tumor model featuring adoptive tumor-infiltrating lymphocyte (TIL) therapy was used. Tumor growth control (p < 0.001) and survival analyses suggest that adenovirus was most effective in enabling T cell therapy. The complete response rate was 62% for TILs + adenovirus versus 17.5% for TILs + PBS. Of note, TIL biodistribution did not explain efficacy differences between viruses. Instead, immunostimulatory shifts in the tumor microenvironment mirrored efficacy results. Overall, the use of oncolytic viruses can improve the utility of T cell therapies, and additional virus engineering by arming with transgenes can provide further antitumor effects. This phenomenon was seen when an unarmed oncolytic adenovirus was compared to Ad5/3-E2F-d24-hTNFa-IRES-hIL2 (TILT-123). A clinical trial is ongoing, where patients receiving TIL treatment also receive TILT-123 (ClinicalTrials.gov: NCT04217473)., Graphical Abstract

Clarithromycin (CLA), a macrolide antibiotic, has been frequently detected in the global surface waters. Concerns have been raised over the potential impacts of CLA on the non-target aquatic species, particularly algae acting as the primary producers in the ecosystem. This study therefore evaluated the toxicological effects of CLA at a range of concentration levels (0, 5, 20, 40, 80 μg L−1) on two green algae, Raphidocelis subcapitata (R. subcapitata) and Chlorella vulgaris (C. vulgaris). The algal growth, photosynthetic pigment contents, lipid peroxidation biomarker malondialdehyde (MDA), responses of antioxidants including superoxide dismutase (SOD), catalase (CAT), glutathione (GSH), glutathione peroxidase (GP), and glutathione S-transferase (GST) were measured. After 7 d exposure, the growth of R. subcapitata was inhibited with the CLA exposure levels higher than 20 μg L−1, whereas the inhibition in C. vulgaris was detected at the concentration level of 80 μg L−1. The MDA contents in both species were elevated. To cope with the increased levels of ROS, the activities of enzymatic antioxidants (SOD, CAT, GP, and GST) and the content of non-enzymatic antioxidant (GSH) in R. subcapitata were all enhanced. However, in C. vulgaris, enhancement was detected only in the activities of antioxidant enzymes (SOD, CAT, and GP). In addition, chlorophyll a, b, and carotenoid contents were all significantly increased in R. subcapitata but decreased in C. vulgaris. The results suggested that R. subcapitata is more sensitive to CLA exposure than C. vulgaris. This study provides insights into the CLA - oxidative stress process in two algae.

In this issue of Developmental Cell, Marques et al. (2019) provide evidence that the main receptor for high-density lipoporoteins (HDL), scavenger receptor B1 (SR-B1), forms large and dynamic homo-multimers at the plasma membrane. This helps the receptor to evade endocytosis, bind HDL, and facilitate selective lipid uptake from HDL.

We compare physiological responses of the crustacean copepod Calanus pacificus and pelagic pteropod mollusk Limacina helicina to ocean temperatures and pH by measuring biomarkers of oxidative stress, antioxidant defences, and the activity of the respiratory electron transport system in organisms collected on the 2016 West Coast Ocean Acidification cruise in the California Current System. Copepods and pteropods exhibited strong but divergent responses in the same habitat; copepods had higher oxygen-reactive absorbance capacity, glutathione-S-transferase, and total glutathione content. The ratio between reduced to oxidised glutathione was higher in copepods than in pteropods, indicating lower oxidative stress in copepods. Pteropods showed higher activities of glutathione reductase, catalase, and lipid peroxidation, indicating increased antioxidant defences and oxidative stress. Thus, the antioxidant defence system of the copepods has a greater capacity to respond to oxidative stress, while pteropods already face severe stress and show limited capacity to deal with further changes. The results suggest that copepods have higher adaptive potential, owing to their stronger vertical migration behaviour and efficient glutathione metabolism, whereas pteropods run the risk of oxidative stress and mortality under high CO2 conditions. Our results provide a unique dataset and evidence of stress-inducing mechanisms behind pteropod ocean acidification responses.

In this work, we studied the effects of residual stresses on the testing of a cracked lap shear specimen (CLS). The analysis focused on tungsten-CFRP laminates for radiation protection applications. A 3-D finite element model was generated and the virtual crack closure technique (VCCT) was applied to determine fracture toughness. Crack propagation was analyzed using cohesive zone modelling (CZM) to combine multiple strain gauge data from experiments and the information from the VCCT. Our analysis indicated that residual stresses cannot be ignored in a valid analysis of tungsten-CFRP hybrid laminates—the experimentation with hybrid CLS specimens requires numerical crack simulation.

Nitrogen (N) availability restricts plant carbon assimilation from the atmosphere in N-limited boreal ecosystems. Boreal forest floor is covered with ericoid plants and in soil, ectomycorrhizal and ericoid mycorrhizal plant roots and fungal hyphae are intermixed. How different mycorrhizal plants affect N mobilization from soil organic matter (SOM) has yet to be elucidated. Here we compared the effects of ericoid plants ( Calluna vulgaris , Vaccinium myrtillus , Vaccinium vitis-idaea ), forming ericoid mycorrhiza (ERM), on soil N chemistry, plant N uptake and SOM decomposition and compared them with ectomycorrhizal (ECM) Pinus sylvestris in a microcosm experiment. Pine and ericoid plants affected soil N similarly, accessing both degradable and recalcitrant N pools. Both ericoid plants and pine took up N from organic source. Ericoid plant roots contained more phenolic compounds and condensed tannins than pine roots which together with lower pH under ericoids suggests substantial changes in soil chemistry and biology. This study underlines the plant stimulation of SOM degradation with special emphasis on ericoid plants. We point out that both ericoid and ectomycorrhizal plants and their interactions should be of interest when N uptake and SOM decomposition in boreal forests are studied.

The majority of nitrogen (N) in boreal forest soils is bound to soil organic matter (SOM) in forms not readily available to plants. Northern boreal forest ecosystems are often N limited, despite atmospheric N deposition, and the utilization of organic N from SOM is of crucial importance to the site productivity. The effect of microbial produced oxidative SOM degrading enzymes (laccase and manganese peroxidases) and proteases on soil N forms and availability was studied in a pot experiment with or without a Scots pine (Pinus sylvestris L.) seedling. The combination of SOM degrading enzymes and proteases decreased the total soil N content and increased the N losses significantly in the absence of the Scots pine seedlings. The total soil N content also decreased in the presence of the Scots pine seedlings, irrespective of the enzyme treatment. Most of the other N parameters studied were not sensitive to enzyme additions, and differed only between planted and non-planted treatments. Our results show that the alkylamine content of boreal forest soil are at the same levels as nitrate. We showed that SOM decomposition, stimulated by oxidative enzyme additions, is a key step in soil organic N utilization, while proteases alone do not increase N use from SOM. Plants stimulate N losses from SOM highlighting the importance of rhizosphere processes in soil C and N cycling.

This study describes workers' exposure to fine and ultrafine particles in the production chain of ferrochromium and stainless steel during sintering, ferrochromium smelting, stainless steel melting, and hot and cold rolling operations. Workers' personal exposure to inhalable dust was assessed using IOM sampler with a cellulose acetate filter (AAWP, diameter 25 mm; Millipore, Bedford, MA). Filter sampling methods were used to measure particle mass concentrations in fixed locations. Particle number concentrations and size distributions were examined using an SMPS+C sequential mobile particle sizer and counter (series 5.400, Grimm Aerosol Technik, Ainring, Germany), and a hand-held condensation particle counter (CPC, model 3007, TSI Incorporated, MN). The structure and elemental composition of particles were analyzed using TEM-EDXA (TEM: JEM-1220, JEOL, Tokyo, Japan; EDXA: Noran System Six, Thermo Fisher Scientific Inc., Madison,WI). Workers' personal exposure to inhalable dust averaged 1.87, 1.40, 2.34, 0.30, and 0.17 mg m(-3) in sintering plant, ferrochromium smelter, stainless steel melting shop, hot rolling mill, and the cold rolling mill, respectively. Particle number concentrations measured using SMPS+C varied from 58 × 10(3) to 662 × 10(3) cm(-3) in the production areas, whereas concentrations measured using SMPS+C and CPC3007 in control rooms ranged from 24 × 10(3) to 243 × 10(3) cm(-3) and 5.1 × 10(3) to 97 × 10(3) cm(-3), respectively. The elemental composition and the structure of particles in different production phases varied. In the cold-rolling mill non-process particles were abundant. In other sites, chromium and iron originating from ore and recycled steel scrap were the most common elements in the particles studied. Particle mass concentrations were at the same level as that reported earlier. However, particle number measurements showed a high amount of ultrafine particles, especially in sintering, alloy smelting and melting, and tapping operations. Particle number concentration and size distribution measurements provide important information regarding exposure to ultrafine particles, which cannot be seen in particle mass measurements.

C-hordein is a monomeric prolamin protein in barley. The unusual primary structure of C-hordein has highly repetitive sequences and forms a secondary structure of beta-turns. C-hordein structure is similar to that of collagen protein, whose degradation by metal-catalysed oxidation has been intensively studied. No information exists on the metal catalysed oxidation of C-hordein, however. In this study, copper-catalysed hydrogen peroxide induced oxidation of C-hordein caused substantial degradation and formed some insoluble compounds. The use of a gliadin standard in R5 ELISA determinations causes an overestimation of hordeins in a sample. A C-hordein standard was therefore directly used as a standard, thus allowing the C-hordein to be analysed as its oxidised prolamin product. After 48 h of oxidation, the prolamin concentration of oxidised C-hordein decreased to 20% of its original amount for competitive ELISA, and to 3% for sandwich ELISA methods. Carbonyl groups were formed during the oxidation. Backbone fragmentation and side-chain modification suggested structural changes of R5 epitopes in C-hordein. Oxidation is an alternative to enzymatic hydrolysis when degrading and modifying C-hordein.

This work aimed at predicting the toxic effects of phenolic compounds in Ba River on the health of female sharpbelly (Hemiculter lucidus) by the de novo transcriptomic analysis of the liver. Sharpbelly, a native fish living in freshwater ecosystem of East Asia, were sampled upstream, near, and downstream of a wastewater discharge to the Ba river. Based on the occurrence of bisphenol A (BPA), nonylphenol (NP), and 4-tert-octylphenol (4-t-OP) in the water and fish sampled from each site, up-, mid-, and down-stream were interpreted as control, high, and low treatment groups, respectively. In the mid-stream group the Fulton's condition factor (CF) and body weight were remarkably increased by approximate 20%; the gonado-somatic index (GSI) and hepatosomatic index (HSI) in mid-stream fish showed a similar increasing trend but lacking of statistical difference. Exposure to wastewater effluent caused 160 and 162 differentially expressed genes (DEGs) in up-mid and down-mid stream groups, respectively. Two sets of DEGs were primarily enriched in the signaling pathways of drug metabolism, endocrine system, cellular process, and lipid metabolism in the mid-stream sharpbelly, which may alter the fish behavior, disrupt the reproductive function, and lead to hypothyroidism, hepatic steatosis, etc. Taken together, our results linked the disrupted signaling pathways with activities of phenolic compounds to predict the potential effects of wastewater effluent on the health of wild fish.

Methotrexate polyglutamates (MTXpg) facilitate incorporation of thioguanine nucleotides into DNA (DNA-TG, the primary cytotoxic thiopurine metabolite and outcome determinant in MTX/6-mercaptopurine treatment of childhood ALL). We hypothesized that mapping erythrocyte levels of MTXpg with 1–6 glutamates and their associations with DNA-TG formation would facilitate future guidelines for maintenance therapy dosing. Summed MTX with 1–6 glutamates resolved by LCMS [median (interquartile): 5.47 (3.58–7.69) nmol/mmol hemoglobin] was in agreement with total MTX by radio ligand assay. In 16,389 blood samples from 1426 ALL maintenance therapy patients, MTXpg3 21.0 (15.2–27.4)% was the predominant metabolite, and MTXpg1 (the maternal drug) constituted 38.6 (27.2–50.2)% of MTXpg1–6. All subsets correlated; the strongest associations were between metabolites with similar polyglutamate lengths. Correlations of MTXpg1 with MTXpg2 and MTXpg3,4,5,6 were rs = 0.68 and rs = 0.25–0.42, respectively. Intercorrelations of MTXpg3,4,5,6 were all rs ≥ 0.51. MTXpg4 accounted for 29.8 (24.7–33.3)% of MTXpg3–6, yet explained 96% of the summed MTXpg3–6 variation. MTXpg1–4, MTXpg1–6, MTXpg2–6 and MTXpg3 were all associated with DNA-TG levels (p

In this article, we introduce for the first time, a method to manufacture cellulose based electrically conductive non-woven three-dimensional (3D) structures using the foam forming technology. The manufacturing is carried out using a minimum amount of processing steps, materials, and hazardous chemicals. The optimized solution applies a single surfactant type and a single predefined portion for the two main processing steps: (1) the dispersing of nanocellulose (NC) and carbon nanotubes (CNT) and (2) the foam forming process. The final material system has a concentration of the used surfactant that is not only sufficient to form a stable and homogeneous nanoparticle dispersion, but it also results in stable foam in foam forming. In this way, the advantages of the foam forming process can be maximized for this application. The cellulose based composite material has a highly even distribution of CNTs over the NC network, resulting a conductivity level of 7.7 S/m, which increased to the value 8.0 S/m after surfactant removal by acetone washing. Also, the applicability and a design product case &lsquo, Salmiakki&rsquo, were studied where the advantages of the material system were validated for a heating element application.

Understanding the interactive effects of multiple stressors on pelagic mollusks associated with global climate change is especially important in highly productive coastal ecosystems of the upwelling regime, such as the California Current System (CCS). Due to temporal overlap between a marine heatwave, an El Niño event, and springtime intensification of the upwelling, pteropods of the CCS were exposed to co-occurring increased temperature, low Ωar and pH, and deoxygenation. The variability in the natural gradients during NOAA’s WCOA 2016 cruise provided a unique opportunity for synoptic study of chemical and biological interactions. We investigated the effects of in situ multiple drivers and their interactions across cellular, physiological, and population levels. Oxidative stress biomarkers were used to assess pteropods’ cellular status and antioxidant defenses. Low aragonite saturation state (Ωar) is associated with significant activation of oxidative stress biomarkers, as indicated by increased levels of lipid peroxidation (LPX), but the antioxidative activity defense might be insufficient against cellular stress. Thermal stress in combination with low Ωar additively increases the level of LPX toxicity, while food availability can mediate the negative effect. On the physiological level, we found synergistic interaction between low Ωar and deoxygenation and thermal stress (Ωar:T, O2:T). On the population level, temperature was the main driver of abundance distribution, with low Ωar being a strong driver of secondary importance. The additive effects of thermal stress and low Ωar on abundance suggest a negative effect of El Niño at the population level. Our study clearly demonstrates Ωar and temperature are master variables in explaining biological responses, cautioning the use of a single parameter in the statistical analyses. High quantities of polyunsaturated fatty acids are susceptible to oxidative stress because of LPX, resulting in the loss of lipid reserves and structural damage to cell membranes, a potential mechanism explaining extreme pteropod sensitivity to low Ωar. Accumulation of oxidative damage requires metabolic compensation, implying energetic trade-offs under combined thermal and low Ωar and pH stress. Oxidative stress biomarkers can be used as early-warning signal of multiple stressors on the cellular level, thereby providing important new insights into factors that set limits to species’ tolerance to in situ multiple drivers.

Calcium has been proposed to diminish metal toxicity by the modulation of the oxidative stress. This study explores the effects of Ca availability and metal exposure on oxidative stress biomarkers in great tit (Parus major) nestlings. Nests were supplemented with Ca (Ca-supplemented group) or not supplemented (Control group) in a metal-polluted and a background zone in SW Finland. Metal concentrations were analyzed from feces. We analyzed antioxidants (tGSH, GSH:GSSG ratio, CAT, GST, GPx, SOD), protein carbonylation and lipid peroxidation in red cells of nestlings. Ca-supplemented and fast-growing nestlings showed higher CAT activity to cope with reactive oxygen species (ROS) generated during intensive growth and metabolism. SOD and GPx (the latter not statistically significant) were more active in the polluted area, possibly reflecting higher ROS production in nestlings from this zone due to the enhanced metal exposure and smaller size. Antioxidant levels changed over the range of metal concentrations depending on the Ca levels in plasma, suggesting that higher Ca levels stimulate antioxidants and mitigate the impacts of metals. Ca supplementation may improve nestling traits and reproductive output when antioxidants are enhanced in a situation of oxidative challenge. Therefore, Ca should be considered in future studies assessing metal exposure and effects on wild birds.

N-Acylation by lipase A from Candida antarctica (CAL-A) in ethyl butanoate was applied to the kinetic resolution of tert-butyl esters of 3-amino-3-phenylpropanoic acid (E>100), 3-amino-4-methylpentanoic acid (E>100) and 3-aminobutanoic acid (E=60) on 1.0–2.0 m scale. With the N-acylated resolution products, the exceptional ability of CAL-A to hydrolyse amides and bulky tert-butyl esters was then studied. In all N-acylated tert-butyl esters, chemoselectivity favoured the amide bond cleavage. The tert-butyl ester bond was left intact with 3-amino-3-phenylpropanoate, whereas with 3-amino-4-methylpentanoate and 3-aminobutanoate the CAL-A-catalysed hydrolysis of tert-butyl ester followed the amide hydrolysis.

Aquatic organisms face multiple stressors in natural ecosystems. More and more often painkillers are detected in surface waters since their prescription has increased worldwide within the last years. Here we examined the effects of the non-steroidal anti-inflammatory drug (NSAID) diclofenac and hypoxia on three-spined sticklebacks (Gasterosteus aculeatus). We exposed sticklebacks to an environmentally relevant concentration of diclofenac (1μg/L) for 14days, to 24h of hypoxia (2.0mg O2/L), and a combination of both. Hypoxia and diclofenac both can be associated with oxidative stress in fish, but it is unclear whether they would act synergistically. Expression analysis of genes related to antioxidant response, hypoxia response, and chemical metabolism in gills showed that diclofenac alone had little effect, while the combination of hypoxia and diclofenac affected transcript levels most, indicating synergistic effects of these stressors. Of the antioxidant enzymes, only superoxide dismutase activity remained unchanged by treatments, while glutathione peroxidase (GPx) was the most affected antioxidant response on both the transcript and activity levels. Our results suggest that diclofenac may lead to suppressed catalase (CAT) activity but increased GPx activity, probably as compensatory mechanism to remove increasing H2O2 in the gills, and that this response is not affected by hypoxia. The activities of lactate dehydrogenase, CAT, and GPx also showed temporal variability during treatments, which can be attributable to tissue-specific circadian rhythms. Our study shows how responses to NSAIDs and hypoxia can interact in fish, suggesting that getting more insight into temporal variation and about the different levels of regulation of environmental responses is necessary in future studies.

The progress of the conversion, the yield, the structure and the morphology of the produced carbonaceous materials as a function of time were systematically studied with pyrolysis-GC/FID and FESEM microscope. The conversion of galactoglucomannan, bleached kraft pulp and TEMPO oxidized cellulose nanofibrils followed the reaction route of glucose being slower though with fibrous material, higher molar mass and viscosity. The conversion of kraft lignin was minor following completely different reaction route. Carbonaceous particles of different shape and size were produced with yields between 23% and 73% after 4 h with being higher for lignin than carbohydrates. According to the results, potential pulp mill streams represent lignocellulosic resources for generation of carbonaceous materials.

Thiol-ene polymer formulations are raising growing interest as new low-cost fabrication materials for microfluidic devices. This study addresses their feasibility for microchip electrophoresis (MCE) via characterization of the effects of UV curing conditions and aging on the surface charge and wetting properties. A detailed comparison is made between stoichiometric thiol-ene (1:1) and thiol-ene formulations bearing 50% molar excess of allyls ("enes"), both prepared without photoinitiator or other polymer modifiers. Our results show that the surface charge of thiol-ene 1:1 increases along with increasing UV exposure dose until a threshold (here, about 200J/cm(2)), whereas the surface charge of thiol-ene 2:3 decreases as a function of increasing UV dose. However, no significant change in the surface charge upon storage in ambient air was observed over a period of 14 days (independent of the curing conditions). The water contact angles of thiol-ene 2:3 (typically 70-75°) were found to be less dependent on the UV dose and storing time. Instead, water contact angles of thiol-ene 1:1 slightly decrease (from initial 90 to 95° to about 70°) as a function of UV increasing exposure dose and storing time. Most importantly, both thiol-ene formulations remain relatively hydrophilic over extended periods of time, which favors their use in MCE applications. Here, MCE separation of biologically active peptides and selected fluorescent dyes is demonstrated in combination with laser-induced fluorescence detection showing high separation efficiency (theoretical plates 8200 per 4cm for peptides and 1500-2700 per 4cm for fluorescent dyes) and lower limits of detection in the sub-μM (visible range) or low-μM (near-UV range) level.

The crushing strengths of four different experimental magnesium aluminate spinel (MgAl 2 O 4 ) granule beds were monitored with the axial die pressing test after heat treatments. Precursor, magnesium hydroxide (Mg(OH) 2 ) and magnesium oxide (MgO) as Mg precursor and aluminium oxide hydroxide Al(O)OH and α-Al 2 O 3 as Al precursor, were used for experimental granules, which were manufactured via a dispersion manufacturing and spray-drying process. After spray-drying, granules were heat treated in air at 1000, 1100, 1200, 1300 and 1400 °C. In order to understand the potential effect of precursor, phase structure, morphology, particle size distribution and density of granules on crushing strength behaviour, scanning X-ray diffraction (XRD) was used together with electron microscopy (SEM) and laser diffraction (LDPA) for characterisation. All precursor mixtures formed spherical granules during the spray-drying process and pure spinel phase structure during heat treatment. The crushing strength test results indicated that the Al precursor clearly affected the crushing strength behaviour of experimental granule beds. The highest strength was observed for granule beds with Al(O)OH) as Al and Mg(OH) 2 as Mg precursor.

Oat protein produced by a dry milling process was enzymatically deamidated by a food-grade protein-glutaminase (PG), and the effects on structure, solubility and emulsifying properties of oat proteins were studied. The reactions were conducted at neutral pH and low salt concentration conditions. Oat proteins were deamidated up to a deamidation degree of 59%. The solubility of proteins doubled. Oil-in-water emulsions prepared with the native and deamidated oat proteins differed as the emulsions prepared from oat proteins with high deamidation degree had a more uniform oil droplet particle size and longer stability. Fourier transform infrared (FT-IR) analysis demonstrated that oat protein secondary structure became more flexible by deamidation. Protein-glutaminase mediated deamidation appeared to be a promising technique to improve oat protein functionality such as emulsifying ability and solubility.

Self-lubricating Al 2 O 3 +1–10 vol% Mo composites were synthesized by pulsed electric current sintering. Molybdenum was found to strongly influence the coefficient of friction against alumina. Lubricious oxide of Mo 4 O 11 formed within the wear track during wear at 400 °C and reduced the coefficient of friction. The composites with Mo had coefficient of friction of 0.35–0.71 at 25 °C and 0.27–0.36 at 400 °C. No measureable wear was observed for composites with 1, 2.5 and 5 vol% Mo. Nanoscratch tests were performed inside the wear tracks to study the friction properties of the transfer film. Alumina molybdenum composites performed as wear resistant materials with very good friction properties at 400 °C owing formation of Mo 4 O 11 during wear.

The effect of N-substitution was examined for the enantioselective lipase-catalyzed ring-opening reaction of racemic 4-phenylazetidin-2-one with methanol in dry organic solvents. Marked differences in the reactivity of various N-protected 4-phenylazetidin-2-ones were observed. Preparative-scale reactions with Candida antarctica lipase B (Novozym 435 preparation) yielded N-acylated methyl (R)-3-amino-3-phenylpropanoates with enantiomeric excess (ee) values >99 % in up to a 49 % isolated yield, whereas Thermomyces lanuginosus lipase (Lipozyme TM IM) gave enantiomerically enriched methyl (S)-3-acetamido-3-phenylpropanoate. Candida antarctica lipase A catalyzed the cleavage of the N-chloroacetyl protective group, whereas all of the other examined lipases underwent the ring-opening reaction.

Hybrid material systems, such as combinations of tungsten foils and carbon fibre-reinforced plastic (CFRP), are replacing metal alloy concepts in spacecraft enclosures. However, a good adhesion between the tungsten oxide scale and the epoxy resin used is required. Here, the effects of a hydrofluoric–nitric–sulphuric-acid (HFNS) treatment on tungsten oxides and subsequent adhesion to CFRP are analysed using atomic force microscopy (AFM), X-ray photoelectron spectroscopy (XPS) and fracture testing. The work shows that HFNS treatment results in decreased oxygen content, over 50% thinner tungsten trioxide (WO 3 ) layer and increased nano-roughness on thin tungsten foils. Fracture testing established a 39% increase in the average critical strain for tungsten–CFRP specimens after HFNS treatment was carried out on tungsten. The effect of the oxide scale modification regarding the critical strain energy release rate was Δ G c ≈ 8.4 J/m 2 .

International audience; Pollution with low concentrations of pharmaceuticals, especially when combined with low-oxygen conditions (hypoxia), is a threat to aquatic ecosystems worldwide. The non-steroidal anti-inflammatory drug diclofenac is commonly detected in wastewater effluents, and has potential to accumulate in the bile of fish. Diclofenac has been shown to activate aryl hydrocarbon receptor (AHR), which induces transcription in the metabolic enzyme cytochrome P450 1a (cyp1a). Previously, crosstalk has been shown to occur between AHR and hypoxia inducible factor 1 (HIF-1). In addition, both of these transcription factors interact with the proteins regulating circadian (24-h) rhythms in vertebrates. Yet little is known about the significance of these interactions during simultaneous exposure to chemicals and hypoxia in fish in vivo. We exposed wild-caught three-spined sticklebacks (Gasterosteus aculeatus) to diclofenac (1 μg/L, 14 days), hypoxia (2.0 mg/L, up to 24 h) and the combination of both. We then analyzed markers of chemical biotransformation (EROD activity, cyp1a and ahr mRNA levels), glycolysis (lactate dehydrogenase (LDH) enzyme activity, ldh and enolase 1a mRNA levels), and the transcription of core circadian clock genes clock and period 1 in liver tissue. Samples were taken at three time points during the light period in order to address disturbances in the circadian variation of metabolic processes. The results show that mRNA levels and LDH activity tended to be lowest before the dark period, but this pattern was disturbed by hypoxia and diclofenac. Diclofenac and hypoxia co-exposure induced EROD activity more strongly than diclofenac exposure alone, while cyp1a mRNA level was increased also by hypoxia and diclofenac alone. LDH activity and mRNA expression showed a clear time-dependent response during hypoxia, which is consistent with the previously suggested decreased accumulation of HIF-1 during the dark period. Furthermore, LDH activity and transcription was disturbed by diclofenac, indicating important effects of environmental pollutants in disturbing natural acclimation. This study demonstrates the need for more studies to understand the potential disturbances in endogenous rhythms caused by environmental pollution in natural populations.

The concentration of residual barley prolamin (hordein) in gluten-free products is overestimated by the R5 ELISA method when calibrated against the wheat gliadin standard. The reason for this may be that the composition of the gliadin standard is different from the composition of hordeins. This study showed that the recognition of whole hordein by R5 antibody mainly came from C-hordein, which is more reactive than the other hordeins. The proportion of C-hordein in total hordein ranged from 16 to 33% of common Finnish barley cultivars used in this study and was always higher than that of omega-gliadin, the homologous protein class in the gliadin standard, which may account for the overestimation. Thus, a hordein standard is needed for barley prolamin quantification instead of the gliadin standard. When gluten-free oat flour was spiked with barley flour, the prolamin concentration was overestimated 1.8-2.5 times with the gliadin standard, whereas estimates in the correct range were obtained when the standard was 40% C-hordein mixed with an inert protein. A preparative-scale method was developed to isolate and purify C-hordein, and C-hordein is proposed as a reference material to calibrate barley prolamin quantification in R5-based assays.

We have developed a workflow to extract, separate, and semi-quantify bioactive oxysterols from mouse colon tissues and fecal matters using solid- and liquid-phase extractions, enzymatic and chemical modifications, and stable-isotope dilution LC/MS/MS. The method was applied to a dextran sodium sulphate (DSS)-induced mouse colitis model, which revealed that one particular dihydroxycholesterol (diOHC), 7α,25-diOHC, was significantly elevated in both colon tissue and fecal matters of mice with colitis compared to that in naive mice. The extent of 7α,25-diOHC elevation was positively correlated with colitis severity.

Cells store excess lipids as two major compounds, triacylglycerols (TAGs) and cholesteryl esters (CEs), inside lipid droplets (LDs). The degree of lipid ordering is considered to play a major role in the mobility and enzymatic processing of lipids in LDs. Here, we provide evidence that polarized third-harmonic generation (THG) microscopy distinguishes between native TAG- and CE-enriched LDs in cells due to the different ordering of the two lipid species. We first demonstrate that the responses from synthetic TAG- and CE-enriched LDs using THG microscopy with linear and circular polarizations differ according to their different intrinsic ordering. We then employ simulations to dissect how polarization effects influence the THG from an isotropic LD. Finally, we induce TAG- and CE-enriched LDs in murine macrophages and demonstrate that polarized THG responses increase in a nonlinear fashion with increasing CE/TAG ratio. This suggests that with an increasing CE content, there is a rather sharp transition toward increased LD ordering. Our results demonstrate that polarized THG microscopy enables label-free quantitative analysis of LD ordering and discriminates between compositionally different LDs in intact mammalian cells.

Cancer is one of the most common reasons for death in dogs. One promising approach is oncolytic virotherapy. We assessed the oncolytic effect of genetically modified vaccinia viruses in canine cancer cells, in freshly excised tumour biopsies, and in mice harbouring canine tumour xenografts. Tumour transduction efficacy was assessed using virus expressing luciferase or fluorescent marker genes and oncolysis was quantified by a colorimetric cell viability assay. Oncolytic efficacy in vivo was evaluated in a nude mouse xenograft model. Vaccinia virus was shown to infect most tested canine cancer cell lines and primary surgical tumour tissues. Virus infection significantly reduced tumour growth in the xenograft model. Oncolytic vaccinia virus has antitumour effects against canine cancer cells and experimental tumours and is able to replicate in freshly excised patient tumour tissue. Our results suggest that oncolytic vaccinia virus may offer an effective treatment option for otherwise incurable canine tumours.

Burkholderia cepacia lipase (lipase PS-D) catalysed acylation with 3,3-difluoro-4-phenyl-, -thiophen-3-yl- and -4-pyridylazetidin-2-ones was examined for the formation of N-Boc-protected 6-O-acylated sugar–β-amino acid conjugates from methyl α-D-galacto-, -gluco- and mannopyranosides and Boc2O. The 6-O-acylated glycopyranoside–β-amino acid conjugates were isolated and characterized. The low solubility of the gluco- and mannopyranosides and the high reactivity of the pyridylazetidinone restricted product formation. Activation of the β-lactam ring by the presence of fluorine substituents was shown to be necessary for the enzymatic acylation reaction. The (S)-enantiomers of the racemic β-lactam substrates reacted with the sugars.

Oncolytic Western Reserve strain vaccinia virus selective for epidermal growth factor receptor pathway mutations and tumor-associated hypermetabolism was armed with human granulocyte-macrophage colony-stimulating factor (GMCSF) and a tdTomato fluorophore. As the assessment of immunological responses to human transgenes is challenging in the most commonly used animal models, we used immunocompetent Syrian golden hamsters, known to be sensitive to human GMCSF and semipermissive to vaccinia virus. Efficacy was initially tested in vitro on various human and hamster cell lines and oncolytic potency of transgene-carrying viruses was similar to unarmed virus. The hGMCSF-encoding virus was able to completely eradicate subcutaneous pancreatic tumors in hamsters, and to fully protect the animals from subsequent rechallenge with the same tumor. Induction of specific antitumor immunity was also shown by ex vivo co-culture experiments with hamster splenocytes. In addition, histological examination revealed increased infiltration of neutrophils and macrophages in GMCSF-virus-treated tumors. These findings help clarify the mechanism of action of GMCSF-armed vaccinia viruses undergoing clinical trials.