Elevated Concentrations Research Articles

SARS-CoV-2 infection induces hyaluronan production in vitro and hyaluronan levels in COVID-19 patients relate to morbidity and long-term lung impairment: a prospective cohort study

ABSTRACT We previously demonstrated that the lungs of deceased COVID-19 patients were filled with a clear hydrogel consisting of hyaluronan (HA). In this translational study, we investigated the role of HA at all stages of COVID-19 disease to map the consequences of elevated HA on morbidity and identify the mechanism of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-induced HA production. A reduced alveolar surface area was observed in the lungs of deceased COVID-19 patients compared to healthy controls, as visualized by a 3D rendering of lung morphology using light-sheet fluorescence microscopy. We confirmed the presence of HA in lung biopsies and found large quantities of proinflammatory fragmented HA. The association of systemic HA in blood plasma and disease severity was assessed in patients with mild (WHO Clinical Progression Scale, WHO-CPS, 1–5) and severe COVID-19 (WHO-CPS, 6–9) during the acute and convalescent phases and related to lung function. We found that systemic levels of HA were high during acute COVID-19 disease, remained elevated during convalescence, and were associated with a reduced diffusion capacity. In vitro 3D-lung models, differentiated from primary human bronchial epithelial cells, were used to study the effects of SARS-CoV-2 infection on HA metabolism, and transcriptomic analyses revealed a dysregulation of HA synthases and hyaluronidases, both contributing to increased HA in apical secretions. Furthermore, corticosteroid treatment reduced the inflammation and downregulated HA synthases. Our findings demonstrate that HA plays a role in COVID-19 morbidity and that sustained elevated HA concentrations may contribute to long-term respiratory impairment. IMPORTANCE This study provides insights into the role of hyaluronan (HA) in the severity and long-term impact of COVID-19 on lung function. Through extensive morphological examination of lung tissues and a multicenter study, we identified that HA levels are significantly elevated in COVID-19 patients, correlating with a reduced lung diffusion capacity during convalescence. Using a 3D-lung model, we further uncovered how severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2 infection causes a dysregulated HA metabolism, leading to increased HA production. Our findings provide valuable insights into the pathogenesis of SARS-CoV-2 and suggest that targeting HA metabolism could offer new therapeutic avenues for managing COVID-19, particularly to prevent long-term lung impairment. Additionally, HA holds potential as a biomarker for predicting disease severity, which could guide personalized treatment strategies.

Immunoglobulin G4 in primary Sjögren’s syndrome and IgG4-related disease - connections and dissimilarities

Primary Sjögren’s syndrome (pSS) is an autoimmune disease, with B cell hyperactivation and autoantibody production as its immunological hallmarks. Although the distinction between immunoglobulin G4-related disease (IgG4-RD) and pSS, based on the presence or absence of certain autoantibodies, seems easy to make, possibility of elevated serum IgG4 concentration and often similar organ involvement may lead to a misdiagnosis. The increased serum concentration of IgG4 in IgG4-RD is not clearly linked to the pathogenesis of IgG-RD and it has been suggested that it may constitute just an epiphenomenon. The aim of this article is to discuss the presence of IgG4 in pSS and IgG4-RD and its potential significance for these two diseases.

Mycobacterium marinum MMAR_0267-regulated copper utilization facilitates bacterial escape from phagolysosome

The host limits Mycobacterium tuberculosis (Mtb) by enriching copper in high concentrations. This research investigates how Mtb escapes copper stress. The membrane protein encoded by Mtb Rv0102, when its homolog in M. smegmatis (MSMEG_4702) was knocked out, resulted in a fourfold decrease in intracellular copper levels and enhanced tolerance to elevated extracellular copper concentrations. Similarly, knockout mutants of its homolog in M. marinum (MMAR_0267) showed increased virulence in zebrafish and higher bacterial load within macrophages. In THP-1 cells infected with MMAR_0267 deletion mutants, the intracellular survival of these mutants increased, along with reduced THP-1 cell apoptosis. Deficiency in copper down-regulated the transcriptional level of the virulence factor CFP-10 in M. marinum, suppressed cytosolic signaling via the macrophage STING pathway, leading to decreased production of IFN-β and reduced cell apoptosis. In conclusion, these findings highlight the significant impact of copper on the survival and reproduction of mycobacteria, underscoring the importance of studying mycobacterial adaptation mechanisms in copper-rich environments.

Reduction in mucosal-associated invariant T cells (MAIT) in APECED patients is associated with elevated serum IFN-γ concentration.

Mucosal-associated invariant T cells (MAIT) are innate-like lymphocytes enriched in mucosal organs where they contribute to antimicrobial defense. APECED is an inborn error of immunity characterized by immune dysregulation and chronic mucocutaneous candidiasis. Reduction in the frequency of circulating MAITs has been reported in many inborn errors of immunity, but only in a few of them, the functional competence of MAITs has been assessed. Here, we show in a cohort of 24 patients with APECED, that the proportion of circulating MAITs was reduced compared with healthy age and sex-matched controls (1.1% vs. 2.6% of CD3+ T cells; p < 0.001) and the MAIT cell immunophenotype was more activated. Functionally the IFN-γ secretion of patient MAITs after stimulation was comparable to healthy controls. We observed in the patients elevated serum IFN-γ (46.0 vs. 21.1 pg/mL; p = 0.01) and IL-18 (42.6 vs. 13.7 pg/mL; p < 0.001) concentrations. Lower MAIT proportion did not associate with the levels of neutralizing anti-IL-22 or anti-IL-12/23 antibodies but had a clear negative correlation with serum concentrations of IFN-γ, IL-18, and protein C-reactive protein. Our data suggest that reduction of circulating MAITs in patients with APECED correlates with chronic type 1 inflammation but the remaining MAITs are functionally competent.

Comparable glucagon-stimulated amino acid suppression in individuals with and without hepatic steatosis or steatohepatitis.

Hepatic amino acid (AA) metabolism and glucagon secretion are linked in a feedback cycle in which circulating AAs stimulate glucagon secretion, while glucagon stimulates hepatic AA catabolism. It has been proposed that metabolic dysfunction-associated steatotic liver disease (MASLD) leads to hepatic glucagon resistance, which may result in hyperaminoacidemia and hyperglucagonemia. We tested the glucagon effect on AA metabolism in subjects with obesity; 11 with steatohepatitis (MASH), 10 with steatosis (MAS), and seven subjects (CON) without steatosis. We performed a somatostatin clamp with infusions of insulin and low-dose followed by high-dose glucagon. We measured plasma levels of 17 AAs and assessed hepatic fat content (FF%) and body fat distribution (visceral and subcutaneous adipose tissue mass) by MRI. HighGlucagon suppressed plasma total AA equally in all groups; MASH 13% (SD 9%), MAS 14% (7%), CON 11% (5%), respectively. In univariate regression analyses visceral adipose tissue mass (β = 0.471, P = 0.011) and AA concentration at LowGlucagon (β = ─0.524, P = 0.004), but not FF% (β = ─0.243, P = 0.213), were significant predictors of AA reduction. Using a stepwise backward multiple regression approach revealed similar results. Total and specific AA levels (glutamic acid, tyrosine) were higher in both MASLD groups during the study and FF% was positively correlated to a number of individual AAs. Though finding elevated AA concentrations in subjects with MASLD, we conclude that in MASLD patients that do not have elevated glucagon at baseline, glucagon suppresses circulating AA levels equally in subjects with and without MASLD. NCT04042142.

As-deposited and dewetted Cu layers on plasma treated glass: Adhesion study and its effect on biological response

Improving the adhesion of nanosized copper films to a glass substrate is vital for their application in electronics and medicine, as it enhances their overall reliability. For this purpose, we employed Ar plasma etching (240 s) and magnetron sputtering to create copper layers on a glass substrate. Furthermore, we investigated the effect of subsequent solid state dewetting (at 300 °C) of Cu nanolayers on the interface stability. Increasing the sputtering time resulted in elevated copper concentration, UV-Vis absorption, conductivity, and surface roughness. The as-deposited and dewetted samples exhibited very good wettability with water contact angles below 60°. Importantly, plasma treatment improved the adhesion of the Cu layers to the glass. Subsequent dewetting accelerated surface diffusion and the oxidation of Cu atoms, causing structural and morphological changes. The presence of CuO after dewetting caused loss of the surface plasmon resonance (SPR) band in the UV-Vis spectrum and a decrease in sample conductivity due to the transformation of the copper layer from a metal to an oxide. Biological testing revealed a more pronounced bactericidal effect for the as-deposited Cu layer against E. coli and S. epidermidis on contrary to dewetted samples. The similar cytotoxic trend was observed for human dermal fibroblasts and hepatocytes. Nonetheless, biological testing confirmed better cell adhesion on dewetted Cu layers compared to the as-deposited ones. Therefore, our copper nanostructured samples could find application as antibacterial coatings of biomedical devices.

Plasma fatty acid levels and risk of non-small cell lung cancer: a large-scale prospective cohort study

BackgroundNon-small cell lung cancer (NSCLC) ranks among the most prevalent and lethal malignancies globally. Fatty acids (FAs) play a significant role in diverse physiological and pathological mechanisms, yet their precise involvement in NSCLC remains poorly understood.MethodsThis study utilized a large-scale prospective cohort of 249,132 participants, observed over an average of 12 years, to investigate the relationship between different FAs and NSCLC risk. Analytical approaches included Cox proportional hazards regression, Kaplan–Meier survival analysis, accelerated failure time (AFT) modeling, and restricted cubic spline (RCS) analysis.ResultsDuring the follow-up period, 1,460 participants were diagnosed with NSCLC. Cox regression analysis demonstrated that elevated levels of docosahexaenoic acid (DHA), linoleic acid (LA), and omega-3 were inversely associated with NSCLC risk. Kaplan–Meier curves, along with AFT models, corroborated that elevated concentrations of DHA and LA significantly delayed NSCLC onset. Additionally, RCS analysis uncovered nuanced dose–response relationships between these FAs and NSCLC. Stratified analyses highlighted variability based on smoking status, gender, and body mass index subgroups.ConclusionThe concentration of specific FAs exhibits a significant association with NSCLC risk. These results offer a foundation for devising dietary FA composition adjustments aimed at reducing NSCLC risk.

Effects of Gossypetin on Glucose Homeostasis in Diet-Induced Pre-Diabetic Rats

Natural flavonoids exert many potential health benefits, including anti-hyperglycaemic effects. However, the effects of gossypetin (GTIN) on glucose homeostasis in pre-diabetes have not yet been investigated. This study examined the effects of GTIN on key markers of glucose homeostasis in a diet-induced pre-diabetic rat model. Pre-diabetes was induced by allowing the animals to feed on a high-fat high-carbohydrate (HFHC) diet supplemented with 15% fructose water for 20 weeks. Following pre-diabetes induction, the pre-diabetic animals were sub-divided into five groups (n = 6), where they were either orally treated with GTIN (15 mg/kg) or metformin (MET) (500 mg/kg), both with and without dietary intervention, over a 12-week period. The results demonstrated that animals in the untreated pre-diabetic (PD) control group exhibited significantly higher fasting and postprandial blood glucose levels, as well as elevated plasma insulin concentrations and increased homeostatic model assessment for insulin resistance (HOMA2-IR) index, relative to the non-pre-diabetic (NPD) group. Similarly, increased caloric intake, body weight and plasma ghrelin levels were observed in the PD control group. Notably, these parameters were significantly reduced in the PD animals receiving GTIN treatment. Additionally, glycogen levels in the liver and skeletal muscle, which were disturbed in the PD control group, showed significant improvement in both GTIN-treated groups. These findings may suggest that GTIN administration, with or without dietary modifications, may offer therapeutic benefits in ameliorating glucose homeostasis disturbances associated with the PD state.

Recombinant neorudin and its active metabolite hirudin: the fate in vivo of a novel anticoagulant drug

Thrombosis, a prevalent condition, can provoke severe health issues like acute coronary syndrome (ACS), deep vein thrombosis (DVT), and pulmonary embolism (PE). The rising incidence of these diseases annually significantly impacts patient wellbeing and poses a substantial burden on healthcare systems. Recombinant neorudin is a developing anticoagulant drug for thrombotic diseases whose phase I clinical trials has been completed. The distribution pattern of it and its active metabolite, hirudin, in thrombi, blood surrounding the thrombus and peripheral blood remains uncertain. This study explored their distribution using a rat arteriovenous bypass thrombosis model, revealing higher neorudin levels in blood surrounding the thrombus and elevated hirudin concentrations in thrombus. Recombinant neorudin significantly increased Thrombin Time (TT) in both plasma surrounding the thrombus and peripheral blood, and reduced the wet weight of the thrombus. The results above demonstrated the anticoagulant and antithrombotic efficacy of recombinant neorudin in vivo. Give the distribution pattern of neorudin and hirudin, we hypothesized that neorudin was cleaved at the site of thrombus formation to produce hirudin, leading to the rapid accumulation of hirudin within local thrombi and resulting in a higher concentration inside the thrombus. This insight was crucial for understanding the action mechanisms of anticoagulants in thrombosis management and provided a valuable guidance for therapeutic strategies in treating thrombotic diseases.

Prebiotic inulin controls Th17 cells mediated central nervous system autoimmunity through modulating the gut microbiota and short chain fatty acids

ABSTRACT Multiple sclerosis (MS) is an autoimmune disease characterized by inflammatory demyelination occurring in the central nervous system (CNS). Inulin is a common prebiotic that can improve metabolic disorders by modulating the gut microbiota. However, its capacity to affect CNS autoimmunity is poorly recognized. Experimental autoimmune encephalomyelitis (EAE) is a classical mouse model of MS. Herein, we found that oral administration of inulin ameliorated the severity EAE in mice, accompanied by reductions in inflammatory cell infiltration and demyelination in the CNS. These reductions were associated with decreased proportion and numbers of Th17 cells in brain and spleen. Consistent with the findings, the serum concentrations of IL-17, IL-6, and TNF-α were reduced in inulin treated EAE mice. Moreover, the proliferation of auto-reactive lymphocytes, against MOG35-55 antigen, was attenuated ex vivo. Mechanistically, inulin treatment altered the composition of gut microbiota. It increased Lactobacillus and Dubosiella whereas decreased g_Prevotellaceae_NK3B31_group at the genus level, alongside with elevated concentration of butyric acid in fecal content and serum. In vitro, butyrate, but not inulin, could inhibit the activation of MOG35-55 stimulated lymphocytes. Furthermore, fecal microbiota transplantation assay confirmed that fecal contents of inulin-treated normal mice had an ameliorative effect on EAE mice. In contrast, antibiotic cocktail (ABX) treatment diminished the therapeutic effect of inulin in EAE mice as well as the reduction of Th17 cells, while supplementation with Lactobacillus reuteri restored the amelioration effect. These results confirmed that the attenuation of inulin on Th17 cells and inflammatory demyelination in EAE mice was dependent on its modulation on gut microbiota and metabolites. Our findings provide a potential therapeutic regimen for prebiotic inulin supplementation in patients with multiple sclerosis.

Systematic investigation on bastnaesite flotation in the presence of dissolved calcium ions using organic acids as chelating agents

This study systematically investigated the potential of organic acids as Ca2+ ions chelating agents in bastnaesite flotation. Naturally, bastnaesite is associated with calcium-bearing gangue minerals, such as calcite (CaCO3) and fluorite (CaF2). The flotation beneficiation of bastnaesite in the presence of calcium-bearing minerals (e.g., calcite) is exceptionally complex due to the release of Ca2+ ions into the flotation pulp which changes the solution chemistry and bastnaesite surface properties. This study seeks to address the challenge of dissolved Ca2+ ions on bastnaesite flotation using organic acids, namely lactic, succinic, and citric acids. Systematic micro-flotation experiments were followed by electro-kinetic tests, solution chemistry studies, and XPS characterization. The results of this study showed that the elevated concentration of Ca2+ ions dramatically decreased bastnaesite flotation recovery values. However, organic acids formed soluble chelates with Ca2+ ions, effectively eliminating the detrimental impact of dissolved Ca2+ ions on bastnaesite flotation.

Cardiac biomarker concentrations in cats with elevated pancreatic lipase concentrations.

Cardiac biomarker concentrations are elevated in dogs with pancreatitis, but it is unknown if this is also the case for cats. The serum feline pancreatic lipase immunoreactivity (fPLI) of serum samples from 60 cats was quantified using the Spec fPL assay. Serum N-terminal pro-B-type natriuretic peptide (NT-proBNP) and high-sensitivity cardiac troponin I (HST) concentrations were also measured using commercial assays. Five of 20 cats with fPLI concentrations of 8.8mg/L or more had above reference interval NT-proBNP concentrations. Serum NT-proBNP concentrations were not correlated with fPLI concentrations (p = 0.31) and were not different in each diagnostic category for fPLI results (p = 0.24). Serum HST concentrations were positively correlated with fPLI concentrations (r = 0.278, 95% confidence interval: 0.02‒0.50, p = 0.03). However, they were not significantly different in each diagnostic category for fPLI results (p = 0.16). Unidentified co-variates could contribute to the association between fPLI and cardiac biomarker concentrations. Cats with elevated fPLI concentrations may have elevated cardiac biomarker concentrations.

Fibrinolytic Dysfunction and Endotheliopathy After Major Thermal Injury: Considerations Needed for New Approaches to Burn Shock Resuscitation.

In recent years, it has become apparent that fibrinolytic dysfunction and endotheliopathy develop in up to 40% of patients during the first hours following thermal injury and are associated with poor outcomes and increased resuscitation requirements. Rapidly following burn injury, the fibrinolytic system is activated, with activation generally greater with increased severity of injury. Very high plasma concentrations of plasmin-antiplasmin complex (marker of activation), have been associated with mortality. Patients display hyperfibrinolytic, physiologic/normal or hypofibrinolytic/fibrinolytic shutdown phenotypes, as assessed by viscoelastic assay. Phenotypes change in over 50% of patients during the acute burn resuscitation period, with some patterns (maladaptive) associated with increased mortality risk and others (adaptive, trending toward the physiologic phenotype) associated with survival. Endotheliopathy, as reflected in elevated plasma concentrations of syndecan-1 has also been associated with increased mortality. Here we review the incidence and effects of these responses after burn injury and explore mechanisms and potential interactions with the early inflammatory response. Available data from burn and non-burn trauma suggest that the fibrinolytic, endothelial, and inflammatory systems interact extensively and that dysregulation in one may exacerbate dysregulation in the others. This raises the possibility that successful treatment of one may favorably impact the others.

Profiling of metabolic dysregulation in ovarian cancer tissues and biofluids

Ovarian cancer (OC) is the most lethal gynecologic cancer, mainly due to late diagnosis with widespread peritoneal spread at first presentation. We performed metabolomic analyses of ovarian and paired control tissues using capillary electrophoresis-mass spectrometry and liquid chromatography-mass spectrometry to understand its metabolomic dysregulation. Of the 130 quantified metabolites, 96 metabolites of glycometabolism, including glycolysis, tricarboxylic acid cycles, urea cycles, and one-carbon metabolites, showed significant differences between the samples. To evaluate the local and systemic metabolomic differences in OC, we also analyzed low or non-invasively available biofluids, including plasma, urine, and saliva collected from patients with OC and benign gynecological diseases. All biofluids and tissue samples showed consistently elevated concentrations of N1,N12-diacetylspermine compared to controls. Four metabolites, polyamines, and betaine, were significantly and consistently elevated in both plasma and tissue samples. These data indicate that plasma metabolic dysregulation, which the most reflected by those of OC tissues. Our metabolomic profiles contribute to our understanding of metabolomic abnormalities in OC and their effects on biofluids.

Hydrogen-natural gas fuel blending in a “rich burn” engine with 3-way catalyst

Interest in hydrogen (H2) fuels is growing, with industry planning to produce it with stranded or excess energy from renewable sources in the future. Natural gas (NG) utility companies are now taking action to blend H2 into their preexisting pipelines to reduce greenhouse gas (GHG) emissions from burning NG. Stoichiometric (“rich burn”) NG engines that operate on pipeline NG and will receive blended fuel as more gas utilities expand H2 production. These engines are typically chosen for their low emissions owing to the 3-way catalyst control, so the focus of this paper is on the change in emissions like carbon monoxide (CO) and nitrogen oxides (NOx) as the fuel is blended with up to 30% H2 by volume. The Caterpillar CG137-8 natural gas engine used for testing was originally designed for industrial gas compression applications and is a good representative for most “rich burn” engines used across industry for applications such as power generation, gas compression, and water pumping. A significant greenhouse gas (GHG) emissions reduction is observed as more H2 is added to the fuel. Increasing H2 in the fuel changes combustion behavior in the cylinder, resulting in faster ignition and higher cylinder pressures, which increase engine-out NOx emissions. Post-catalyst CO and NOx both decrease slightly with increasing H2 while operating at the optimal “air-fuel” equivalence ratio (λ). A “rich burn” engine with 3-way catalyst can tolerate up to 30% H2 (by vol.) while still meeting NOx and CO emissions limits. However, at elevated levels of H2, increased engine-out NOx emissions narrow the λ range of operation. As H2 is added to NG pipelines, some “rich burn” engine systems may require larger catalysts or more precise λ control to accommodate the increased NOx production associated with a H2-NG blend. Sudden step-increases in H2 cause dramatic changes in λ, resulting in large emissions of post-catalyst NOx during the transition. Comparable changes in H2 at elevated concentrations cause larger spikes in NOx than at lower concentrations. Better tuned engine controllers respond more quickly and produce less NOx during H2 step-transitions.

Uranium Speciation and Mobilization in Thawing Permafrost.

Uranium is a toxic and pervasive geogenic contaminant often associated with organic matter. Its abundance and speciation in organic-rich permafrost soils are unknown, thereby limiting our ability to assess risks associated with uranium mobilization during permafrost thaw. In this study, we assessed uranium speciation in permafrost soil and porewater liberated during thaw using active-layer and permafrost samples from a study area in Yukon, Canada where elevated uranium concentrations occur in bedrock and groundwater. Permafrost contained 1.1-28 wt % organic carbon and elevated uranium (range 7.6-1040 μg g-1, median 25 μg g-1) relative to local bedrock. The highest soil uranium concentrations were encountered in catchments hosting uranium-enriched bedrock and correlated positively with soil organic carbon. X-ray absorption spectroscopy, micro-X-ray fluorescence, and electron microscopy analyses revealed that solid-phase uranium predominantly occurs as uranium(VI) associated with soil organic matter. Extended X-ray absorption fine structure (EXAFS) analyses suggested the presence of uranium(VI) coordinated with carbon, consistent with bidentate-mononuclear uranyl complexation on carboxyl groups. Permafrost thaw produced circumneutral pH porewater (pH 6.2-7.5) with elevated dissolved uranium (0.5-203 μg L-1). Geochemical modeling indicated that calcium-uranyl-carbonate complexes dominated the dissolved uranium speciation. This study highlights that permafrost soil can mobilize uranium upon thaw and that uranium fate is linked to dynamic biogeochemical reactions involving organic carbon and groundwater chemistry.

Decreased dust particles amplify the cloud cooling effect by regulating cloud ice formation over the Tibetan Plateau.

Ice-nucleating particles (INPs) can initiate cloud ice formation, influencing cloud radiative effects (CRE) and climate. However, the knowledge of INP sources, concentrations, and their impact on CRE over the Tibetan Plateau (TP)-a highly climate-sensitive region-remains largely hypothetical. Here, we integrated data from multisource satellite observations and snowpack samples collected from five glaciers to demonstrate that dust particles constitute primary INP sources over the TP. The springtime dust influxes lead to seasonally elevated ice concentrations in mixed-phase clouds. Furthermore, the decadal reduction in dustiness from 2007 to 2019 results in decreased springtime dust INPs, thereby amplifying the cooling effect of clouds over the TP, with a 1.98 ± 0.39-watt per square meter reduction in surface net CRE corresponding to a 0.01 decrease in dust optical depth. Our findings elucidate previously unidentified pathways of climate feedback from an atmospheric INP perspective, especially highlighting the crucial role of dust in aerosol-cloud interactions.

Sensitivity of montane grassland water fluxes to warming and elevated CO2 from local to catchment scale: A case study from the Austrian Alps

Study region: Montane grassland within the Gulling catchment, Austrian Alps. Study focus: A climate-change experiment in a grassland ecosystem used lysimeters and HYDRUS-1D models to quantify changes in evapotranspiration (ET) and groundwater recharge (GWR) due to warming (＋3 °C) and elevated CO2 concentrations (ΔCO2; ＋300 ppm). Findings at the plot-scale were generalized and transferred to the surrounding catchment, half comprised of grassland, using three lumped rainfall–runoff models and two spatially-distributed Community Water Models, differing in soil hydraulic properties.New hydrological insights for the region: Warming increased ET and decreased GWR and river discharge compared to ambient conditions. ΔCO2 increased stomatal resistance, which partially offset warming effects. In scenarios combining warming and ΔCO2, the impact of warming was higher than ΔCO2 effect. Elevation influenced the sensitivity of ET to warming, which was greater at the catchment scale than at the plot scale, while GWR was more sensitive to warming at the plot scale. Under dry conditions, GWR and discharge exhibited increased sensitivity to warming at both scales. HYDRUS-1D successfully reproduced lysimeter experiment results and their sensitivity to warming and ΔCO2. Despite model agreement on water flux sensitivity to climate changes, the varying response magnitudes highlight the need for a multi-model approach in climate impact assessments. This study provides insights into how climate change might impact hydrological dynamics of montane grassland systems across the Central European Alps.

The effect of extracellular polymeric substances on the distribution and transmission of antibiotic resistance genes treating antibiotic wastewater via microbial electrolysis cells

Microbial electrolysis cells (MEC) have emerged as a prominent technology for the treatment of antibiotics-containing wastewater in recent years. However, there remains a dearth of comprehensive exploration regarding the influence of extracellular polymers substances (EPS) on the distribution and transmission of antibiotic resistance genes (ARGs) in MEC. In this study, we quantified the distribution of ARGs in MEC by Fluorescence quantitative polymerase chain reaction and explored with emphasis on impact of EPS component on ARGs transmission at under different concentrations of roxithromycin. Results showed that the absolute abundance of ARGs in the electrode biofilm was 1 - 2 orders of magnitude higher than that in the anolyte. Specifically, EPS-associated ARGs accounted for 2.31% - 11.18% of ARGs in electrode biofilm. The presence of elevated roxithromycin concentration led to electroactive microorganisms (Geobacter and Geothrix) as potential hosts of ARGs. In addition, both protein and polysaccharide content in the electrode biofilm increased with increasing roxithromycin concentration and showed positive correlations with EPS-associated ARGs. Fluorescence quenching experiments further elucidated that tryptophan and tyrosine residues in EPS could bind to ARGs effectively, contributing the hindering the ARGs transmission between hosts. Therefore, increased EPS content within electrode biofilm could reduce the concentration of ARGs present in anolyte while also influencing ARGs distribution throughout MEC. This study provides valuable insights into the distribution of ARGs in MEC systems and the role of EPS in regulating ARGs migration.

Effect of oral or subcutaneous administration of cyanocobalamin in hypocobalaminemic cats with chronic gastrointestinal disease or exocrine pancreatic insufficiency.

No prospective study has evaluated the efficacy of oral supplementation with cobalamin in hypocobalaminemic cats. To investigate the efficacy of oral or SC supplementation with cyanocobalamin in normalizing serum cobalamin and methylmalonic acid (MMA) concentrations in hypocobalaminemic cats with chronic gastrointestinal disease (CGID) or exocrine pancreatic insufficiency (EPI). Forty-eight client-owned hypocobalaminemic (<290 ng/L) cats with normal or abnormally high serum MMA concentrations. This study was conducted based on the prospective randomized clinical trial method. Cats with CGID or EPI were randomly assigned to 2 groups that received either oral or SC supplementation with cobalamin (250 μg/cat) for 12 and 10 weeks, respectively, in addition to other medical and dietary interventions. Each cat was evaluated 3 times (baseline, 6-week postsupplementation, and 1-week postcompletion) by measuring serum cobalamin and MMA concentrations. In cats with CGID or EPI, cobalamin concentrations were normalized in all cats that received either oral or SC supplementation (mean 100% [95% CI: 80.6%-100%] in both groups in cats with CGID and 100% [67.6%-100%] in both groups in cats with EPI). Among 37 cats with elevated MMA concentrations at baseline (21 cats with CGID and 16 cats with EPI), MMA concentrations were normalized in most cats with CGID (70% in oral and 82% in SC group) or EPI (88% in both groups). In hypocobalaminemic cats with CGID or EPI, in conjunction with other medical and dietary interventions, both oral and SC supplementation are effective at normalizing serum cobalamin and MMA concentrations.