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Gene therapy (GT) has rapidly attracted renewed interest as a treatment for otherwise incurable diseases, with several GT products already on the market and many more entering clinical testing for selected indications. Clonal tracking techniques based on vector integration enable monitoring of the fate of engineered cells in the blood of patients receiving GT and allow assessment of the safety and efficacy of these procedures. However, owing to the limited number of cells that can be tested and the impracticality of studying cells residing in peripheral organs without performing invasive biopsies, this approach provides only a partial snapshot of the clonal repertoire and dynamics of genetically modified cells and reduces the predictive power as a safety readout. In this study, we developed liquid biopsy integration site sequencing, or LiBIS-seq, a polymerase chain reaction technique optimized to quantitatively retrieve vector integration sites from cell-free DNA released into the bloodstream by dying cells residing in several tissues. This approach enabled longitudinal monitoring of in vivo liver-directed GT and clonal tracking in patients receiving hematopoietic stem cell GT, improving our understanding of the clonal composition and turnover of genetically modified cells in solid tissues and, in contrast to conventional analyses based only on circulating blood cells, enabling earlier detection of vector-marked clones that are aberrantly expanding in peripheral tissues.

International audience; Two types of bicyclic N-cyclopropyl glycine ester derivatives have been prepared and put under scrutiny as possible precursors of azomethine ylides. The results demonstrate that they can indeed participate in 1,3-dipolar cycloaddition reactions with dipolarophiles, as illustrated in the cases of phenyl vinyl sulfone, N-phenylmaleimide, diethyl fumarate and diethyl maleate. The relative configurations of the major diastereoisomers produced are consistent with the predicted generation of azomethine ylide species, reacting in concerted cycloaddition processes. This unprecedented way of generating such 1,3-dipoles provides access to functionalised pyrrolizidine and pyrrolidine derivatives, that would be difficult to make directly by more classic methods. It was also found that using phenyl vinyl sulfone or N-phenylmaleimide as the dipolarophile reactant, a domino nucleophilic conjugate addition/1,3-dipolar cycloaddition process may operate competitively.

International audience; In tetrahydrofuran, the electrochemical reduction of Cp2TiIVCl2 (2 mM) generated three titanium(III) complexes which were in equilibrium: [Cp2TiCl2]•−, [Cp2TiCl]• and (Cp2TiCl)2. Although the anion radical [Cp2TiCl2]•− was the main species produced under these conditions, cyclic voltammetry investigations clearly showed that the proportion of the three electrogenerated TiIII complexes can be modified as a function of the amounts of chloride ion present in the solution. Accordingly, the presence of Mg2+ ions, which led to the consumption of chloride ions through the formation of MgCl2, favoured the formation of [Cp2TiCl]• and, consequently, of the corresponding dimer (Cp2TiCl)2. The electrochemical behaviours of Cp2TiIVCl2 and of the electrogenerated low-valent Ti complexes were also investigated in the presence of amide and alkyne derivatives. Under these conditions, titanium complexes could not only interact with the amide carbonyl group, but also with the alkyne triple bond, provided the latter was not sterically hindered. Interestingly, the carbonyl group and the triple bond had antagonist effects on redox properties of titanium(III) complexes.

Gram-negative bacteria possess an asymmetric outer membrane (OM) composed primarily of lipopolysaccharides (LPSs) on the outer leaflet and phospholipids (PLs) on the inner leaflet. The loss of this asymmetry due to mutations in the LPS biosynthesis or transport pathways causes the externalization of PLs to the outer leaflet of the OM and leads to OM permeability defects. Here, we used metabolic labeling to detect a compromised OM in intact bacteria. Phosphatidylcholine synthase expression in Escherichia coli allowed for the incorporation of exogenous propargylcholine into phosphatidyl(propargyl)choline and exogenous 1-azidoethyl-choline (AECho) into phosphatidyl(azidoethyl)choline (AEPC), as confirmed by LC/MS analyses. A fluorescent copper-free click reagent poorly labeled AEPC in intact wild-type cells but readily labeled AEPC from lysed cells. Fluorescence microscopy and flow cytometry analyses confirmed the absence of significant AEPC labeling from intact wild-type E. coli strains and revealed significant AEPC labeling in an E. coli LPS transport mutant (lptD4213) and an LPS biosynthesis mutant (E. coli lpxC101). Our results suggest that metabolic PL labeling with AECho is a promising tool for detecting a compromised bacterial OM, revealing aberrant PL externalization, and identifying or characterizing novel cell-active inhibitors of LPS biosynthesis or transport.­

Soil stores over 2500 Pg carbon (C), with the majority of C stored in deep soil layers (> 30 cm). Soil C can be lost to the atmosphere when organic compounds are mineralized to carbon dioxide (CO2, via oxidative decay or respiration) and moved upward through the soil profile (via diffusion). Soil moisture status can influence the balance between respiration and diffusion, thereby altering the soil CO2 concentration and flux. However, it is unclear how wetting and drying influence soil CO2 dynamics in surface vs. deep soil layers. Thus, we irrigated three soil profiles in Mediterranean arable land and continuously monitored soil CO2 concentration at 15, 30, 50, 70 and 90 cm during wetting and drying phases under ambient temperature conditions. We estimated gas diffusivity, CO2 flux, and temperature responses of soil CO2 concentration during the experiment. Decreases in gas diffusivity during the wetting period coincided with increases in soil CO2 concentrations. However, the negative gas diffusivity-soil CO2 concentration relationship did not hold true all the time, implying that CO2 production was the driving factor for the apparent soil CO2 concentration. We observed hysteretic responses of soil CO2 concentration to temperature as soil moisture varied, with deeper soil CO2 concentration being more sensitive to temperature than surface soil CO2 concentration, especially during the drying phase. The movement of CO2 was upward at all depths during the ambient phase, but the direction and the magnitude of CO2 fluxes varied across the depth gradient during the wetting and drying phases. This study highlights that the relative contribution of gas diffusivity vs. CO2 production to soil CO2 concentration changes with wetting and drying, and that the responses of soil CO2 concentration to temperature are dependent on the antecedent environmental conditions. Also, the downward movement of CO2 during the wetting and drying phases suggests that quantifying surface soil CO2 efflux may underestimate dynamic C processes in deeper soils.

The tumor suppressor gene, p53, is inactivated in the human hepatocellular carcinoma cells line, Hep3B. Berberine has been reported to inhibit the proliferation of cancer cells. This study examined whether apoptosis was induced in berberine-treated Hep3B cells and observed the association between apoptosis and the expression of p53 and DNA methyltransferase (DNMT). The cell viability was measured using an MTT assay. Apoptosis of Hep3B was measured using annexin V flow cytometry. Berberine-treated cells were examined for their DNMT enzymatic activity, mRNA expression, and protein synthesis. The p53 levels were examined by Western blot analysis. The berberine treatment resulted in increased Hep3B cell death and apoptosis in a time- and dose-dependent manner. The DNMT3b activity, mRNA expression, and protein levels all decreased after the berberine treatment. In contrast, the p53 protein levels increased with a concomitant decrease in DNMT3b. No change in the expression of ERK was observed, but the P-ERK levels decreased in a dose dependent manner. These results indicate that a treatment of Hep3B cells with berberine can reduce the expression of DNMT3b, leading to an increase in the tumor suppressant gene p53 and an increase in cell apoptosis. This shows that berberine can effectively suppress the proliferation of liver cancer cells.

Germline RUNX1 mutations underlie a syndrome, RUNX1-familial platelet disorder (RUNX1-FPD), characterized by bleeding symptoms that result from quantitative and/or qualitative defect in platelets and a significantly increased risk for developing hematologic malignancies. Myeloid neoplasms are the most commonly diagnosed hematologic malignancies, followed by lymphoid malignancies of T-cell origin. Here, we describe the first 2 cases of B-cell acute lymphoblastic leukemia (B-ALL) in patients with confirmed germline RUNX1 mutations. While 1 of the patients had a known diagnosis of RUNX1-FPD with a RUNX1 p.P240Hfs mutation, the other was the index patient of a kindred with a novel RUNX1 variant, RUNX1 c.587C>T (p.T196I), noted on a targeted genetic testing of the B-ALL diagnostic sample. We discuss the clinical course, treatment approaches, and the outcome for the 2 patients. Additionally, we describe transient resolution of the mild thrombocytopenia and bleeding symptoms during therapy, as well as the finding of clonal hematopoiesis with a TET2 mutant clone in 1 of the patients. It is critical to consider testing for germline RUNX1 mutations in patients presenting with B-ALL who have a personal or family history of thrombocytopenia, bleeding symptoms, or RUNX1 variants identified on genetic testing at diagnosis.

International audience; Herein, we report a multistep synthesis of novel (meth)acrylate monomers based on gallic acid (GA), a biosourced phenolic acid. The objective of this work was to obtain bio-based polymers exhibiting antioxidant properties provided by monomers derived from gallic acid. The phenolic groups of GA, which are responsible for antioxidant properties, need to be protected for two reasons. On the one hand, functionalization to transform GA into polymerizable monomers must not take place at the phenolic groups because they must remain free to maintain the maximum antioxidant activity in the final polymers. On the other hand, their protection is necessary to prevent radical scavenging during the radical polymerization. After synthesis of such monomers, protected GA-based polymers were thus produced through a photo-mediated RAFT polymerization at room temperature by evaluating two trithiocarbonate-type chain transfer agents (CTAs). The kinetics and molecular weight distributions were studied depending on the monomers and the CTAs. Protected polymers were then deprotected to afford polymeric chains carrying one free gallic acid moiety on each monomer unit. The antioxidant activity of these free GA-based polymers was demonstrated either through the DPPH free radical scavenging property or through the inhibition of methyl linoleate oxidation.

This study aimed to extend the shelf-life of coffee-containing milk beverage by adding Theobroma cacao (cacao nibs) extract. To prepare the beverage sample containing cacao nibs extract, 0.8% cacao nibs hydrothermal extract was aseptically injected. Qualitative changes in the beverage samples, including antioxidant effect, peroxide value (POV), caffeine content, and sensory parameters were monitored regularly during storage at 10°C, 20°C, and 30°C for 4 wk. The inclusion of cacao nibs extract produced higher antioxidant activity compared to the control. As the storage temperature increased, the POV of all samples increased. Samples with cacao nibs extract generally displayed lower POV than the control. The caffeine content of all samples tended to decrease during storage, with the decrease accentuated by higher storage temperatures. In the shelf-life prediction using the Arrhenius model, the kinetic regressions of the cacao nibs extract-added sample and control were YPOV=1.2212X–2.1141 (r2=0.9713) and YPOV=1.8075X–2.0189 (r2=0.9883), respectively. Finally, the predicted shelf-life of cacao nibs-added group and control to reach the quality limit (20 meq/kg POV) were approximately 18.11 and 12.18 wk, respectively. The results collectively indicate that the addition of cacao nibs extract extends the shelf-life of the coffee-containing milk beverage and heightened the antioxidant effect.

This study aimed to extend the retention of flavor in coffee-containing milk beverage by microencapsulation. The core material was caramel flavor, and the primary and secondary coating materials were medium-chain triglyceride and maltodextrin, respectively. Polyglycerol polyricinoleate was used as the primary emulsifier, and the secondary emulsifier was polyoxyethylene sorbitan monolaurate. Response surface methodology was employed to determine optimum microencapsulation conditions, and headspace solid-phase microextraction was used to detect the caramel flavor during storage. The microencapsulation yield of the caramel flavor increased as the ratio of primary to secondary coating material increased. The optimum ratio of core to primary coating material for the water-in-oil (W/O) phase was 1:9, and that of the W/O phase to the secondary coating material was also 1:9. Microencapsulation yield was observed to be approximately 93.43%. In case of in vitro release behavior, the release rate of the capsules in the simulated gastric environment was feeble; however, the release rate in the simulated intestinal environment rapidly increased within 30 min, and nearly 70% of the core material was released within 120 min. The caramel flavor-supplemented beverage sample exhibited an exponential degradation in its flavor components. However, microcapsules containing flavor samples showed sustained flavor release compared to caramel flavor-filled samples under higher storage temperatures. In conclusion, the addition of coffee flavor microcapsules to coffee-containing milk beverages effectively extended the retention of the coffee flavor during the storage period.

International audience; Dextran-covered poly(D,L-lactide) (PLA) nano-and microparticles were prepared by emulsion/solvent evaporation (or diffusion) process with the aim of encapsulating alkyl gallates (AGs). In the first step, a solution of PLA and AG in ethyl acetate was emulsified into an aqueous phase containing a hydrophobically modified dextran which acted as stabilizer. The second step consisted in solvent evaporation or diffusion. Emulsification conditions were varied which allowed preparing nano-and microparticle suspensions covering a wide range of surface-average particle diameters between 0.1 µm (in the case of sonication) and 500 µm (in the case of stirring with magnetic barrel) with narrow and reproducible size distributions. Continuous microfluidic emulsification in a flow focusing system led to well-defined microparticles, within the 10-50 µm range. Particles loaded with octyl gallate (OG) and nonyl gallate (NG) were obtained with the three processes and we showed that the encapsulation efficiency of OG and NG varied significantly according to emulsification process. The effect of particle size on the mechanism of in vitro release of encapsulated AGs was investigated. According to the pH of external medium, the kinetics of release was controlled either by Fickian diffusion within solid core or by swelling and hydrolytic degradation of PLA matrix.

BACKGROUND: Pathogen inactivation and cold or cryopreservation of platelets (PLTs) both significantly affect PLT function. It is not known how PLTs function when both are combined. STUDY DESIGN AND METHODS: Standard PLT concentrates (PCs) were compared to pathogen-inactivated PCs treated with amotosalen photochemical treatment (AS-PCT) when stored at room (RT, 22 degrees C), cold (4 degrees C, n = 6), or cryopreservation (-80 degrees C, n = 8) temperatures. The impact of alternative storage methods on both arms was studied in flow cytometry, light transmittance aggregometry, and hemostasis in collagen-coated microfluidic flow chambers. RESULTS Platelet aggregation of cold-stored AS-PCT PLTs was 44% +/- 11% compared to 57% +/- 14% for cold-stored standard PLTs and 58% +/- 21% for RT-stored AS-PCT PLTs. Integrin activation of cold-stored AS-PCT PLTs was 53% +/- 9% compared to 77% +/- 6% for cold-stored standard PLTs and 69% +/- 13% for RT-stored AS-PCT PLTs. Coagulation of cold-stored AS-PCT PLTs started faster under flow (836 +/- 140 sec) compared to cold-stored standard PLTs (960 +/- 192 sec) and RT-stored AS-PCT PLTs (1134 +/- 220 sec). Fibrin formation rate under flow was also highest for cold-stored AS-PCT PLTs. This was in line with thrombin generation in static conditions because cold-stored AS-PCT PLTs generated 297 +/- 47 nmol/L thrombin compared to 159 +/- 33 nmol/L for cold-stored standard PLTs and 83 +/- 25 nmol/L for RT-stored AS-PCT PLTs. So despite decreased PLT activation and aggregation, cold storage of AS-PCT PLTs promoted coagulation. PLT aggregation of cryopreserved AS-PCT PLTs (23% +/- 10%) was not significantly different from cryopreserved standard PLTs (25% +/- 8%). CONCLUSION: This study shows that cold storage of AS-PCT PLTs further affects PLT activation and aggregation but promotes (pro)coagulation. Increased procoagulation was not observed after cryopreservation.

International audience

Iron is an indispensable metabolic cofactor in both pro- and eukaryotes, which engenders a natural competition for the metal between bacterial pathogens and their human or animal hosts. Bacteria secrete siderophores that extract Fe(3+) from tissues, fluids, cells, and proteins; the ligand gated porins of the Gram-negative bacterial outer membrane actively acquire the resulting ferric siderophores, as well as other iron-containing molecules like heme. Conversely, eukaryotic hosts combat bacterial iron scavenging by sequestering Fe(3+) in binding proteins and ferritin. The variety of iron uptake systems in Gram-negative bacterial pathogens illustrates a range of chemical and biochemical mechanisms that facilitate microbial pathogenesis. This document attempts to summarize and understand these processes, to guide discovery of immunological or chemical interventions that may thwart infectious disease.

In the present study, innovative doxorubicin-loaded nanoparticles (NPs) made of a photosensitive poly(o-nitrobenzyl acrylate) (PNBA) hydrophobic matrix and an hydrophilic dextran (Dex) shell were f...

Anthropogenic activities, particularly fertilisation, have resulted in significant increases in reactive nitrogen (rN) in soils globally, leading to eutrophication, acidification, poor air quality, and emissions of the important greenhouse gas N2O. Understanding the partitioning of rN losses into different environmental compartments is critical to mitigate negative impacts, however, loss pathways are poorly quantified, and potential changes driven by climate warming and societal shifts are highly uncertain. We present a coupled soil-atmosphere isotope model (IsoTONE; ISOtopic Tracing Of Nitrogen in the Environment) to partition rN losses into leaching, harvest, NH3 volatilization, and production of NO, N2 and N2O based on a global dataset of soil δ15N, as well as numerous other geoclimatic and experimental datasets. The model was optimized in a Bayesian framework using a time series of N2O mixing ratios and isotopic compositions since the preindustrial era, as well as a global dataset of N2O emission factors (EF). The posterior model results showed that the total anthropogenic flux in 2020 (7.8 Tg N2O-N a-1) was dominated by indirect emissions resulting from N deposition, while the growth rate and trend in anthropogenic N2O was driven by both direct N fertilisation and deposition inputs. In contrast, inputs from fixation N drive natural N2O emissions, and were responsible for subdecadal interannual variability in total emissions.Total N gas (N2O + NO + N2) production and N2O losses were strongly dependent on geoclimate and thus spatially variable, therefore the spatial pattern of N inputs strongly impacted resulting EFs and total N2O emissions. The area-weighted global EF for N2O was 1% of anthropogenic N inputs in 2020, similar to the current IPCC default of 1.4%, however the N input-weighted global EF was 4.3%. Shifts in fertilisation inputs from the temperate Northern hemisphere towards warmer regions with higher EFs such as India and China have led to accelerating N2O emissions (1.02±0.7 Tg N2O-N a-1). In addition, N2O emissions have increased over the past decades due to climate warming (0.76±0.4 Tg N2O-N a-1). Predicted increases in fertilisation in India and Africa in the coming decades could further accelerate N2O-driven climate warming, unless mitigation measures are implemented to increase fertiliser N use efficiency and reduce N2O emission factors.

Simple Summary Odor emission from swine production facilities can irritate the people living in surrounding areas, although the farmers consider odor emission as a part of farming practice. Despite the governmental and institutional efforts, odor-related complaints from the neighborhood communities around the swine farms are rapidly increasing and have been identified as a key concern to sustaining progress of the swine industry globally. Bio-liquor circulation systems (BCSs) in swine farms have become popular among the farmers as an odor reduction technology in Korea. However, due to the lack of appropriate operating strategies, the odor reduction capacity of BCSs is often depleted. In this lab-scale study, a real-time control strategy based on oxidation–reduction potential (ORP) and pH (mV) time profiles was developed and applied for BCS operation. This study shows the potential effectiveness of using ORP and pH (mV) time profiles as operational parameters for the BCS to improve swine manure properties in slurry pits and thus reduce odor emission. Abstract In this study, an attempt was made to develop a real-time control strategy using oxidation–reduction potential (ORP) and pH (mV) time profiles for the efficient operation of bio-liquor circulation system (BCS) in swine farms and its effectiveness in reducing odor emission through improving manure properties in the slurry pit was evaluated. The lab-scale BCS used in this study comprised a bioreactor and a slurry pit. The bioreactor was operated in a sequence of inflow of swine manure → anoxic phase → aerobic phase → circulation to the slurry pit. The improvement in swine manure properties was elucidated by comparing the results of the BCS slurry pit (circulation type, CT) and conventional slurry pit (non-circulation type, NCT). The results revealed that the ORP time profile successfully detected the nitrate knee point (NKP) in the anoxic phase. However, it was less stable in detecting the nitrogen break point (NBP) in the aerobic phase. The pH (mV) time profile showed a more efficient detection of NBP. Compared to the NCT slurry pit, concentrations of ammonium nitrogen (NH4-N) and soluble total organic carbon (STOC) and other analyzed swine manure properties were much lower in the CT slurry pit. In the aspect of odor reduction, around 98.3% of NH3 was removed in the CT slurry pit. The real-time controlled BCS can overcome the drawbacks of fixed time-based BCS operation and therefore can be considered as a useful tool to reduce odor emission from intensive swine farming operations. However, further studies and refinement in control algorithms might be required prior to its large-scale application.

The recycling of organic waste in biogas plants is proposed as a measure to close nutrient cycles and possibly reduce nitrogen losses such as nitrous oxide emissions and nitrate leaching. Ammonia volatilization after fertilizer spreading is yet another nitrogen loss pathway which is often understudied and not yet fully understood but the knowledge is needed in order to optimize fertilizer management. We therefore aimed to quantify the volatilization of ammonia after the trail-hose application of digestates compared to cattle slurry. We hypothesize that digestates have larger and longer lasting nitrogen losses via ammonia volatilization due to higher NH4+ contents and pH values compared to fresh manure. In this project, digested and un-digested organic fertilizers were applied twice per year in a 2.5-years field experiment with three consecutive arable crops (maize, winter wheat and winter barley) under organic farming. We used Automated Low Cost Impinger Systems to measure ammonia emissions after fertilizer application. The emissions were then modeled using the backwards Langrangian stochastic dispersal model with respect to wind conditions. A preliminary presentation of the data indicates that ammonia emissions from the cattle slurry, slurry-based digestate, and industrial digestate are alternately higher or lower. In 2018, emissions from cattle slurry tended to be lower than those from slurry-based digestate and industrial digestate, while in 2019 and 2020 all three liquid organic fertilizers had similar emissions. In the measurement period after the second fertilizer application in 2018, which took place at the end of May, conspicuously high emissions were measured. This can be explained by the high temperatures during this period. Adaptive strategies in fertilizer management should thus consider reduced inputs of organic fertilizers during warm periods.

In this study, the technical, economic and environmental attributes of a full-scale nutrient recovery process connected to the centralized swine wastewater treatment facility (CSWTF) were evaluated. The performance of the process was assessed by introducing influent to the recovery reactor from different components of the CSWTF such as sedimentation tank (swine wastewater) and biological treatment reactor (biologically oxidized material and supernatant of the biologically oxidized material). The results of technical performance assessment revealed that the O–P recovery (87.1–90.7%) and NH4–N removal (66.9–72.1%) efficiencies from the influent of biological treatment reactor were significantly higher than the influent from sedimentation tank (81.7 and 19.8%, respectively, p

The agricultural sector is considered one of the major sources of greenhouse gas (GHG) emissions globally. The livestock industry as a significant contributor, is accounting for about 18% of GHG emissions measured in carbon dioxide (CO2) equivalent from agricultural practices. Depending on farming practices and climatic conditions, GHGs such as methane (CH4) and nitrous oxide (N2O) emissions from livestock agriculture can vary significantly. Country-specific emission factors are, therefore, needed for a precise estimation of GHG emissions and to avoid uncertainties. This study was aimed at estimating the CH4 and N2O emission fluxes from Hanwoo (the most famous and popular Korean native cattle) manure management systems. CH4 and N2O emission fluxes from litter in the Hanwoo cattle barn and composting lot were monitored and calculated for 52 weeks using the dynamic chamber method. The calculated monthly average fluxes of CH4 and N2O from litter in the cattle barn ranged from 0.0 to 30.0 &plusmn, 13.7 and 0.896 &plusmn, 0.557 to 2.925 &plusmn, 2.853 &mu, g/m2 s, respectively during the whole measurement period. While during the composting period, the monthly average of CH4 and N2O emission fluxes were varied from 1.449 &plusmn, 0.783 to 86.930 &plusmn, 19.092 and 0.511 &plusmn, 0.410 to 2.629 &plusmn, 1.105 &mu, g/m2 s, respectively. The calculated emission fluxes of CH4 and N2O from manure management systems in this study were almost 5.4 and 2.1 times, respectively higher than the values reported for the Asian, South and North American countries in the 2006 Intergovernmental Panel on Climate Change (IPCC) Guidelines for National Greenhouse Gas Inventories. Overall, this study initiates the process along with signifies the importance of developing country-specific GHG inventories for the effective reduction of GHG emissions from the livestock sector in Korea.

International audience; Necrotizing pneumonia induced by Panton-Valentine leukocidin–secreting Staphylococcus aureus is a rare but life-threatening infection that has been described in patients after they had influenza. We report a fatal case of this superinfection in a young adult who had coronavirus disease.

International audience; Vascular dysfunction is an essential element found in many cardiovascular pathologies and in pathologies that have a cardiovascular impact such as chronic kidney disease (CKD). Alteration of vasomotricity is due to an imbalance between the production of relaxing and contracting factors. In addition to becoming a determining factor in pathophysiological alterations, vascular dysfunction constitutes the first step in the development of atherosclerosis plaques or vascular calcifications. In patients with CKD, alteration of vasomotricity tends to emerge as being a new, less conventional, risk factor. CKD is characterized by the accumulation of uremic toxins (UTs) such as phosphate, para-cresyl sulfate, indoxyl sulfate, and FGF23 and, consequently, the deleterious role of UTs on vascular dysfunction has been explored. This accumulation of UTs is associated with systemic alterations including inflammation, oxidative stress, and the decrease of nitric oxide production. The present review proposes to summarize our current knowledge of the mechanisms by which UTs induce vascular dysfunction.

Struvite crystallization using fluidized bed reactors (FBRs) is one of the most commonly used methods for nutrient recovery from different waste streams. However, struvite recovery from swine wastewater containing much higher solids using FBR has not been studied extensively. In this study, we therefore designed and optimized the key operating conditions parameters, i.e., pH (9.0, 9.5, and 10.0), circulation rate (CR) (1.5, 3.0, and 4.5 L/Lreactor&middot, h), and hydraulic retention time (HRT) (1, 3, and 5 h) of FBR to ensure efficient nutrient removal and struvite crystallization from swine wastewater using response surface methodology (RSM) with central composite design (CCD) as experimental design. A magnesium/phosphorus (Mg/P) molar ratio of 1.3 was maintained with MgCl2 according to ortho-phosphate (O-P) concentration of influent and an air diffuser was set to supply air with 0.03 L air/Lreactor&middot, min. The O-P recovery efficiency of over 91% was achieved through the entire runs. Among the operational parameters, pH did not show any significant effect on NH4-N recovery, particle size, and struvite production rate (SPR). The optimal CR over 2.94 L/Lreactor&middot, h was found to be appropriate for efficient removal of nutrients and struvite crystallization. While optimizing the HRT, priority of the process operation such as the production of larger struvite particles or increased struvite productivity should be considered. Therefore, the optimal operational parameters of pH 9.0, CR &gt, 2.94 L/Lreactor&middot, h, and HRT of 1 or 5 h were chosen to obtain better responses through RSM analyses. The findings of this study would be useful in designing and operating either pilot- or full-scale FBR for struvite crystallization from swine wastewater.

3-Deoxy-d-manno-oct-2-ulosonic acid (Kdo) is an essential component of lipopolysaccharides (LPS) in the Gram-negative bacterial outer membrane. Metabolic labeling of Escherichia coli LPS with 8-azido-3,8-dideoxy-d-manno-oct-2-ulosonic acid (Kdo-N3 ) has been reported but is inefficient. For optimization, it is important to understand how exogenous Kdo-N3 enters the cytoplasm. Based on similarities between Kdo and sialic acids, we proposed and verified that the sialic acid transporter NanT imports exogenous Kdo-N3 into E. coli. We demonstrated that E. coli ΔnanT were not labeled with Kdo-N3 , while expression of NanT in the ΔnanT mutant restored Kdo-N3 incorporation. Induced NanT expression in a strain lacking Kdo biosynthesis led to higher exogenous Kdo incorporation and restoration of full-length core-LPS, suggesting that NanT also transports Kdo. While Kdo-N3 incorporation was observed in strains having NanT, it was not detected in Pseudomonas aeruginosa and Acinetobacter baumannii, which lack nanT. However, heterologous expression of E. coli NanT in P. aeruginosa enabled Kdo-N3 incorporation and labeling, though this led to abnormal morphology and growth arrest. NanT seems to define which bacteria can be labeled with Kdo-N3 , provides opportunities to enhance Kdo-N3 labeling efficiency and spectrum, and raises the possibility of Kdo biosynthetic bypass where exogenous Kdo is present, perhaps even in vivo.

For the sediments of the central and southern North Sea different sources of alkalinity generation are quantified by a regional modelling system for the period 2000–2014. For this purpose a formerly global ocean sediment model coupled with a pelagic ecosystem model is adapted to shelf sea dynamics, where much larger turnover rates than in the open and deep ocean occur. To track alkalinity changes due to different nitrogen-related processes, the open ocean sediment model was extended by the state variables particulate organic nitrogen (PON) and ammonium. Directly measured alkalinity fluxes and those derived from Ra isotope flux observation from the sediment into the pelagic are reproduced by the model system, but calcite building and calcite dissolution are underestimated. Both fluxes cancel out in terms of alkalinity generation and consumption. Other simulated processes altering alkalinity in the sediment, like net sulfate reduction, denitrification, nitrification, and aerobic degradation, are quantified and compare well with corresponding fluxes derived from observations. Most of these fluxes exhibit a strong positive gradient from the open North Sea to the coast, where large rivers drain nutrients and organic matter. Atmospheric nitrogen deposition also shows a positive gradient from the open sea towards land and supports alkalinity generation in the sediments. An additional source of spatial variability is introduced by the use of a 3-D heterogenous porosity field. Due to realistic porosity variations (0.3–0.5) the alkalinity fluxes vary by about 4 %. The strongest impact on interannual variations of alkalinity fluxes is exhibited by the temporal varying nitrogen inputs from large rivers directly governing the nitrate concentrations in the coastal bottom water, thus providing nitrate necessary for benthic denitrification. Over the time investigated the alkalinity effluxes decrease due to the decrease in the nitrogen supply by the rivers.

Polysaccharide coated nanoparticles represent a promising class of environmentally friendly latex to replace those stabilized by small toxic molecular surfactants. We report here an in situ formulation of free-surfactant core/shell nanoparticles latex consisting of dextran-based diblock amphiphilic copolymers. The synthesis of copolymers and the immediate latex formulation were performed directly in water using a photo-initiated reversible addition fragmentation chain transfer-mediated polymerization induced self-assembly strategy. A hydrophilic macromolecular chain transfer-bearing photosensitive thiocarbonylthio group (eDexCTA) was first prepared by a modification of the reducing chain end of dextran in two steps: (i) reductive amination by ethylenediamine in the presence of sodium cyanoborohydride, (ii) then introduction of CTA by amidation reaction. Latex nanoparticles were then formulated in situ by chain-extending eDexCTA using 2-hydroxypropyl methacrylate (HPMA) under 365 nm irradiation, leading to amphiphilic dextran-b-poly(2-hydroxypropyl methacrylate) diblock copolymers (DHX). Solid concentration (SC) and the average degree of polymerization - Xnˉ- of PHPMA block (X) were varied to investigate their impact on the size and the morphology of latex nanoparticles termed here SCDHX. Light scattering and transmission electron microscopy analysis revealed that SCDHX form exclusively spherical nano-objects. However, the size of nano-objects, ranging from 20 nm to 240 nm, increases according to PHPMA block length.

3-Deoxy-d-manno-oct-2-ulosonic acid (Kdo) is an essential component of LPS in the outer leaflet of the Gram-negative bacterial outer membrane. Although labeling of Escherichia coli with the chemical reporter 8-azido-3,8-dideoxy-d-manno-oct-2-ulosonic acid (Kdo-N3) has been reported, its incorporation into LPS has not been directly shown. We have now verified Kdo-N3 incorporation into E. coli LPS at the molecular level. Using microscopy and PAGE analysis, we show that Kdo-N3 is localized to the outer membrane and specifically incorporates into rough and deep-rough LPS. In an E. coli strain lacking endogenous Kdo biosynthesis, supplementation with exogenous Kdo restored full-length core-LPS, which suggests that the Kdo biosynthetic pathways might not be essential in vivo in the presence of sufficient exogenous Kdo. In contrast, exogenous Kdo-N3 only restored a small fraction of core LPS with the majority incorporated into truncated LPS. The truncated LPS were identified as Kdo-N3–lipid IVA and (Kdo-N3)2–lipid IVA by MS analysis. The low level of Kdo-N3 incorporation could be partly explained by a 6-fold reduction in the specificity constant of the CMP-Kdo synthetase KdsB with Kdo-N3 compared with Kdo. These results indicate that the azido moiety in Kdo-N3 interferes with its utilization and may limit its utility as a tracer of LPS biosynthesis and transport in E. coli. We propose that our findings will be helpful for researchers using Kdo and its chemical derivatives for investigating LPS biosynthesis, transport, and assembly in Gram-negative bacteria.

We estimate the additional transient surface warming ΔTs caused by a potential reduction of marine dimethyl sulfide (DMS) production due to ocean acidification under the high-emission scenario RCP8.5 until the year 2200. Since we use a fully coupled Earth system model, our results include a range of feedbacks, such as the response of marine DMS production to the additional changes in temperature and sea ice cover. Our results are broadly consistent with the findings of a previous study that employed an offline model set-up. Assuming a medium (strong) sensitivity of DMS production to pH, we find an additional transient global warming of 0.30 K (0.47 K) towards the end of the 22nd century when DMS emissions are reduced by 7.3 Tg S yr−1 or 31 % (11.5 Tg S yr−1 or 48 %). The main mechanism behind the additional warming is a reduction of cloud albedo, but a change in shortwave radiative fluxes under clear-sky conditions due to reduced sulfate aerosol load also contributes significantly. We find an approximately linear relationship between reduction of DMS emissions and changes in top of the atmosphere radiative fluxes as well as changes in surface temperature for the range of DMS emissions considered here. For example, global average Ts changes by −0. 041 K per 1 Tg S yr−1 change in sea–air DMS fluxes. The additional warming in our model has a pronounced asymmetry between northern and southern high latitudes. It is largest over the Antarctic continent, where the additional temperature increase of 0.56 K (0.89 K) is almost twice the global average. We find that feedbacks are small on the global scale due to opposing regional contributions. The most pronounced feedback is found for the Southern Ocean, where we estimate that the additional climate change enhances sea–air DMS fluxes by about 9 % (15 %), which counteracts the reduction due to ocean acidification.

International audience; Alginate-based hydrogel scaffolds are widely used in the field of cartilage regeneration and repair. If the effect of autoclaving on the alginate powder is well known, it is not the same for the possible effects of the sterilization UV treatment on the properties of the hydrogel after polymerization. To select an effective sterilization treatment of alginate-based materials, one must find what are inter-relationship between the characteristics (chemical, physical and mechanical) of alginate-based hydrogel during sterilization, and what consequences have affected on cell behavior. In this study, we investigated the influence of UV sterilization treatments (UV-1 and UV-2: 25 and 50min, respectively) and autoclaving to obtain alginate (Alg)/hyaluronic acid (HA) hydrogel, as well as further evaluated the relationship between physicochemical properties and cell behavior of Alg/HA hydrogel after UVs and autoclaving. The physicochemical properties of this mixture at the powder or polymerized states were analyzed using ATR-FTIR, HPLC-SEC, rheological, indentation testing and sterility testing. The cell behaviors of hydrogels were evaluated by cell viability and proliferation, and chondrogenic differentiation. The effects of treatment parameters and their correlation with the others characteristics were determined statistically by Principal Component Analysis (PCA). In this study, we have shown that the cell behavior in alginate-based hydrogels was not only regulated by physicochemical properties (as molar mass or/and viscosity), but also associated with the controlling of sterilization time. It can provide a basis for choosing an effective method of sterilization, which can keep the mechanical or physical-chemical properties of Alg-based hydrogel scaffold and maintain its cytocompatibility and its ability to induce chondrogenesis from mesenchymal stem cells.

Rivers act as a natural source of greenhouse gases (GHGs) that can be released from the metabolisms of aquatic organisms. Anthropogenic activities can largely alter the chemical composition and microbial communities of rivers, consequently affecting their GHG emissions. To investigate these impacts, we assessed the emissions of CO2, CH4, and N2O from Cuenca urban river system (Ecuador). High variation of the emissions was found among river tributaries that mainly depended on water quality and neighboring landscapes. By using Prati and Oregon Indexes, a clear pattern was observed between water quality and GHG emissions in which the more polluted the sites were, the higher were their emissions. When river water quality deteriorated from acceptable to very heavily polluted, their global warming potential (GWP) increased by ten times. Compared to the average estimated emissions from global streams, rivers with polluted water released almost double the estimated GWP while the proportion increased to ten times for very heavily polluted rivers. Conversely, the GWP of good-water-quality rivers was half of the estimated GWP. Furthermore, surrounding land-use types, i.e. urban, roads, and agriculture, significantly affected the river emissions. The GWP of the sites close to urban areas was four time higher than the GWP of the nature sites while this proportion for the sites close to roads or agricultural areas was triple and double, respectively. Lastly, by applying random forests, we identified dissolved oxygen, ammonium, and flow characteristics as the main important factors to the emissions. Conversely, low impact of organic matter and nitrate concentration suggested a higher role of nitrification than denitrification in producing N2O. These results highlighted the impacts of land-use types on the river emissions via water contamination by sewage discharges and surface runoff. Hence, to estimate of the emissions from global streams, both their quantity and water quality should be included.

Whatever the dairy derived fluids that are heat processed, many studies have demonstrated the key role of beta-lactoglobulin (β-lg) denaturation in the initiation of a dairy fouling and mentioned that interactions between β-lg, phosphate and calcium ions are essential in fouling growth. However only few authors have reported the influence of the phosphate/calcium molar ratio on fouling mechanisms. To partly fill this gap, various whey protein solutions (composed mainly of β-lg) having different phosphate:calcium molar ratios were prepared by rehydrating a whey protein isolate powder and by adding calcium chloride and various amounts of soluble inorganic phosphate (noted Pi). Fouling experiments were carried out at pilot scale using a plate heat exchanger (PHE) at a fixed temperature profile and the deposited mass per channel was measured. Finally, the chemical composition and structure of the proteinaceous fouling layer were observed using an Electron Probe Micro Analyser. Results clearly demonstrated that the phosphate:calcium molar ratio modified the deposit structure and influenced substantially the amount of fouling. These results suggest that the relative proportion of calcium phosphate and calcium ions governs the mechanism of fouling build-up and also sets, to a certain extent, the fouling ability of the whey protein solutions.

Fusarium graminearum is a phytopathogenic fungus that causes Fusarium head blight in small-grain cereals, such as wheat, with significant yield reductions. Moreover, it contaminates the cereal grains with health-threatening mycotoxins, such as deoxynivalenol (DON), jeopardizing food and feed safety. Plant-based biopesticides, i.e. botanicals, have recently gained increased interest in crop protection as alternatives to synthetic chemical products. The main objective of this study was to test the control efficacy of botanicals based on white or Indian/Oriental mustard seed flours (Tillecur – Ti, Pure Yellow Mustard – PYM, Pure Oriental Mustard – POM, Oriental Mustard Bran – OMB) on F. graminearum infection and mycotoxin accumulation in wheat grain. Botanicals at 2% concentration showed a higher efficacy in inhibiting mycelium growth in vitro compared with a prothioconazole fungicide (F). In the growth chamber experiment under controlled conditions, the spraying agents reduced DON content in grain in the following order: F = Ti = PYM > POM > OMB. The antifungal activity of the botanicals may be attributed to their bioactive matrices containing isothiocyanates (ITCs) and phenolic acids. Allyl ITC was detected in POM and OMB at 8.38 and 4.48 mg g–1, while p-hydroxybenzyl ITC was found in Ti and PYM at 2.56 and 2.44 mg g–1, respectively. Considerable amounts of various phenolic acids were detected in all botanicals. Under field conditions, only the use of F significantly decreased F. graminearum infection and DON content in grain. An additional important finding of this study is that disease control was more difficult when infection was done with ascospores than conidia, which might have several potential implications considering that ascospores are more important in Fusarium head blight epidemics. Our results suggest that mustard-based botanicals are promising biopesticides for the control of Fusarium head blight in small-grain cereals, but for field applications, an appropriate formulation is necessary to stabilize and prolong the antifungal activity, especially against ascospores. ISSN:1664-302X

Aiming to prepare oily core pH-sensitive nanocapsules (NCs) for anticancer drugs delivery, the use of a dextran-based transurf (DexN3- τ CTA γ ) as both stabilizer and macromolecular chain transfer agent in methyl methacrylate/2-(diethylamino)ethyl methacrylate (MMA/DEAEMA) miniemulsion copolymerization was investigated. NCs of about 195 nm with an oily-core of Miglyol 810 (M810) and a dextran coverage covalently linked to the poly(MMA-co-DEAEMA) intern shell have been obtained. Compared to the non-sensitive PMMA-based NCs (prepared in a similar way), these novel objects were shown to swell in acidic media and to trigger Coumarin 1 release in physiological relevant pH range. As a starting point of NCs biological effects, cytotoxicity and NCs-proteins interactions studies were performed with both PMMA and poly(MMA-co-DEAEMA)-based NCs. Finally, free azide functions from dextran-based coverage were successfully exploited to attach fluorescent model dyes to NCs surface. The overall results suggest that this novel NCs platform could be potentially used as drug nanocarriers for intravenous injection.

We quantified nitrous oxide (N2O) fluxes and total denitrification (N2O + N2) in an acidic (Ferralsol) and a near-neutral pH soil (Cambisol) to determine whether biochar’s alkalinization effect could be the mechanism inducing potential reductions in N2O fluxes. In Ferralsol, decreases in N2O emissions and in the N2O to N2O + N2 ratio were observed in both biochar and lime treatments. In Cambisol, neither biochar nor lime decreased N2O emissions, despite significantly increasing soil pH. The abundance and community structure of nosZ gene-bearing microorganisms indicated that gene abundances did not explain biochar effects, but a higher diversity of nosZ gene-bearing microorganisms correlated to lower total denitrification. Overall, our results suggest that biochar’s potential to decrease N2O emissions, through soil alkalinization, may be more effective in acidic soils, Soil Systems, 3 (4)

International audience; A multi-reactive polysaccharide-based transurf (acting both as macro-Chain Transfer Agent and stabilizer) was used to confine RAFT polymerization of methyl methacrylate (MMA) at the oil/water (o/w) miniemulsion interface. Dithiobenzoate groups and hydrophobic aliphatic side chains were introduced onto dextran, conferring it both transfer agent properties and ability to stabilize direct miniemulsion of MMA in the presence of a biocompatible oil, used as co-stabilizer. Because of their amphiphilic character, transurfs were initially adsorbed at the (o/w) interface and their reactive sites mediated RAFT polymerization via the R-group approach. PMMA-grafted dextran glycopolymers were consequently produced at the o/w interface, thus leading to dextran coverage/PMMA shell/oily core nanocapsules (NCs) as evidenced by Cryo-TEM analyses. The influence of dextran-based transurf chemistry and oil amount on MMA RAFT polymerization control was investigated. Positive preliminary results on NCs cytotoxicity suggest the potential of these objects for biomedical applications.

Apart from using as fertilizer for plants, the application of struvite may be expanded to animal feed industries through proper pre-treatment. This study aimed to investigate the safety and efficacy of using pre-treated struvite (microwave irradiated struvite (MS) and incinerated struvite (IS)) in animal feeds. For safety assessment, an in vivo toxicity experiment using thirty female Sprague Dawley rats (average body weight (BW) of 200 &plusmn, 10 g) was conducted. The rats were randomly divided into five groups, including a control. Based on the BW, MS and IS were applied daily by oral administration with 1 and 10 mg kg&minus, 1-BW (MS1 and MS10, IS1 and IS10) using dimethyl sulfoxide (DMSO) as a vehicle. A series of jar tests were conducted for four hours to check the solubility of the MS and IS at different pH (pH 2, 4, and 5) and compared to a commercial P source (monocalcium phosphate, MCP, control). The toxicity experiment results showed no significant differences among the treatments in BW and organ (liver, kidney, heart, and lung) weight of rats (p &gt, 0.05). There were no adverse effects on blood parameters and the histopathological examination showed no inflammation in the organ tissues in MS and IS treated groups compared to the control. In an in vitro solubility test, no significant difference was observed in ortho-phosphate (O-P) solubility from the MCP and MS at pH 2 and 4 (p &gt, 0.05), while O-P solubility from MS at pH 5 to 7 was higher than MCP and found to be significantly different (p &lt, 0.05). O-P solubility from IS was the lowest among the treatments and significantly different from MCP and MS in all the experiments (p &lt, 0.05). The results of this study not only suggest that the struvite pre-treated as MS could be a potential alternative source of P in animal feed but also motivate further studies with more stringent designs to better examine the potential of struvite application in diverse fields.

Background Dirofilaria immitis is a filarial parasite of dogs that can cause serious or fatal cardiopulmonary disease. Three studies were conducted to evaluate the efficacy and safety of monthly treatment with moxidectin in a chewable tablet product in combination with sarolaner and pyrantel to prevent heartworm disease in dogs after experimental challenge and in a clinical field study in the USA. Methods In two laboratory studies, dogs (8 per group) that had been inoculated 30 days prior with 50 third-stage D. immitis larvae were randomized to treatment on Day 0 with placebo or combination product, at the minimum dose of 24 µg/kg moxidectin, 2 mg/kg sarolaner and 5 mg/kg pyrantel (as pamoate salt). Study 2 also included groups treated with tablets containing moxidectin-alone (24 µg/kg) or sarolaner-alone (2 mg/kg). Efficacy was evaluated ~ 5 months after inoculation by adult heartworm counts at necropsy. In the field study, 410 dogs ≥ 8 weeks-old from 23 USA veterinary clinics were treated for 11 months with either combination product at 24–48 µg/kg moxidectin, 2–4 mg/kg sarolaner and 5–10 mg/kg pyrantel (n = 272) or Heartgard® Plus (ivermectin/pyrantel) at the label recommended dose rate (n = 138). Efficacy was evaluated on Day 330 using antigen and microfilaria testing to assess adult heartworm infection. Results In the laboratory studies, there were no heartworms recovered from any dog treated with the combination product or moxidectin alone and all dogs treated with placebo or sarolaner-alone were infected with 20–44 adult heartworms. In the field study, all dogs treated with the combination product tested negative for heartworm infection on Day 330, whereas two dogs treated with Heartgard® Plus tested positive. The Heartgard® Plus-treated dogs that tested heartworm positive were from the lower Mississippi River Valley region, where heartworm resistance has been confirmed to occur. The combination product was well tolerated in all studies. Conclusions In laboratory studies, no heartworms were recovered from dogs treated with a single dose of the novel combination product containing moxidectin, sarolaner and pyrantel. Additionally, in the field study no dog tested positive for adult heartworm infection when dosed with the combination product monthly for 11 months, while two dogs treated with Heartgard® Plus tested positive.

Background Moxidectin has previously shown limited efficacy (≤ 44.4%) against confirmed macrocyclic lactone (ML)-resistant Dirofilaria immitis strains at 3 µg/kg after single and multiple oral dosages. Three studies were conducted to evaluate higher oral moxidectin doses for efficacy against confirmed ML-resistant D. immitis strains. Methods Dogs were inoculated with 50 D. immitis L3 and randomly allocated to treatments. Study 1: 6 groups of dogs (n = 8) were inoculated with JYD-34 (Day − 30) and treated as follows: T01, negative control; T02–T05, moxidectin at 3, 6, 12 or 24 µg/kg, respectively, on Day 0 only; T06, moxidectin at 3 µg/kg on Days 0, 30 and 60. Study 2: 10 groups of dogs (n = 5) were inoculated (Day − 30) with either JYD-34 (T01, T03–05) or ZoeLA (T02, T06–T10) and treated as follows: T01 and T02, negative controls; T03–T05, moxidectin at 24, 40 or 60 µg/kg, respectively, on Days 0, 28 and 56; T06 and T09, moxidectin at 3 or 60 µg/kg on Day 0 only; T07, T08 and T10, moxidectin at 24, 40 or 60 µg/kg, respectively, on Days 0, 28 and 56. Study 3: 5 groups of dogs (n = 5) were inoculated with ZoeMO (Day − 28) and treated as follows: T01, negative control; T02, moxidectin at 3 µg/kg moxidectin on Day 0 only; T03–T05, moxidectin at 24, 40 or 60 µg/kg, respectively, on Days 0, 28 and 56. All dogs were necropsied for adult heartworm recovery ~ 4–5 months post-inoculation. Results All moxidectin-treated dogs showed significantly lower worm counts than controls. The efficacy of moxidectin administered once at 3 µg/kg was 19% (JYD-34), 44.4% (ZoeLA) and 82.1% (ZoeMO). Increasing both the dose and the number of dosages of moxidectin improved efficacy, with 100% protection obtained using three dosages of moxidectin at either 40 µg/kg (JYD-34, ZoeMO) or 60 µg/kg (ZoeLA). Three dosages of 24 µg/kg were also highly effective, providing ≥ 98.8% efficacy for all three strains. Conclusions Increasing both the dose and number of consecutive monthly dosages of moxidectin improved the efficacy against ML-resistant heartworms. Based on these data and other technical considerations, the 24 µg/kg dose was considered the optimal dose for further commercial development.

Summary Understanding phosphorus (P) dynamics at the soil aggregate level is necessary to increase P-use efficiency and for reducing reliance on external P inputs. Traditional methods of soil fractionation are often unsuitable for the study of P dynamics within soil aggregates of tropical soil because of the large amounts of strongly P-sorbing iron oxide concretions. Here, we present an alternative method of soil fractionation to assess carbon (C), nitrogen (N) and P cycling in a very weathered tropical soil (total P, 883 ± 19 mg P kg−1) rich in iron concretions (total P, 1136 ± 28 mg P kg−1). This wet-sieving method removes large iron concretions (> 2000 µm) without any marked effect on smaller soil aggregates, which reduces the error associated with overestimation of aggregation. It reduces variation in nutrient content within larger soil aggregates, and also enables nutrient cycling within water-stable soil aggregates to be studied. In addition, because it uses field-moist soil it is possible to measure microbial biomass, community structure, or both, within aggregate fractions. Although this method is important for measuring and understanding C, N and P dynamics within soil aggregate fractions, both the nutrient content and biochemical characteristics of the iron concretions need to be considered to obtain a deeper understanding of nutrient dynamics within the whole soil. Highlights An aggregate fractionation method for tropical soil rich in iron concretions. This method can separate large iron concretions from soil aggregates. Error and variation in aggregation measurements and associated nutrient concentrations are reduced. Measurement of P in concretions and aggregates separately provides a better understanding of P dynamics.

Increasing the input and turnover of root tissue is considered to be one method that may increase carbon (C) inputs and storage in soil. The use of herbicide during pasture renewal (periodic re-sowing of pasture) is expected to increase root inputs and turnover as plants die. The objective of this study was to quantify the short-term impact of pasture renewal on root turnover and C input to soil of ryegrass-clover pastures. Pastures were labelled in the field using a 13C isotope pulse labelling method within 1 m2 clear chambers. Five daily labelling events were carried out during one week in paired treatment plots within 3 replicate paddocks. One plot per paddock was sprayed with herbicide and then the pasture was renewed by direct drilling of seed. The 13C of roots and soil (0–100 mm) was measured at regular intervals over an 89-day period. Herbicide application caused an initial rapid turnover time of 17 days followed by a slower turnover time of 524 days, compared to unsprayed pasture which had a root turnover of 585 days. Faster root turnover following herbicide application resulted in greater cumulative C input to soil over 89 days with approximately double the C input in the sprayed treatment (3238 ± 378 kg C ha−1) compared to the unsprayed treatment (1726 ± 540 kg C ha−1). The use of glyphosate during pasture renewal increased root turnover and resulted in a greater short term cumulative C input to soil. This study provides the first values of root turnover and C input to soil during a pasture renewal event in New Zealand pasture systems and contributes to the understanding of how pasture roots may influence the soil C input following plant death in grassland systems.

Two randomised, blinded, multi-centered field studies were conducted in Europe to demonstrate the efficacy and safety of three monthly oral doses of sarolaner (Simparica™, Zoetis) administered at a minimum dosage of 2.0mg/kg (range 2–4mg/kg) against natural flea or tick infestation of dogs presented as veterinary patients. In the flea study, the improvement in clinical signs associated with flea allergy dermatitis (FAD) was also investigated. The palatability of the sarolaner chewable tablet formulation was evaluated in both studies. Spinosad (Comfortis® Chewable Tablets, Elanco) and fipronil (Frontline® Spot on, Merial) were used as positive controls in the flea and tick study, respectively. Treatments were administered on Days 0, 30 and 60. Efficacy was calculated based on the mean percent reduction of live parasite counts on post-treatment days 14, 30, 60 and 90 versus the pre-treatment count on Day 0. Non-inferiority of sarolaner to the control products was assessed at each time-point using a margin of 15% at the one-sided 0.025 significance level.Dogs were enrolled in a 2:1 ratio (sarolaner:comparator); 285 flea- and 181 tick-infested dogs were assessed for efficacy and safety, and 137 and 48 dogs were assessed for safety only, in the flea and tick study, respectively. There were no treatment-related adverse events.Efficacy against fleas was 98.8%, 99.4%, >99.9% and >99.9% in the sarolaner-treated group and 98.9%, 93.7%, 96.8% and 95.1% in the spinosad-treated group on Days 14, 30, 60 and 90, respectively. Sarolaner was non-inferior to spinosad at all time-points and was superior on Day 30. For the 42 dogs identified as having FAD at enrolment, the clinical signs of FAD improved in all dogs and the incidence was markedly reduced by the end of the study.Efficacy against ticks was 97.4%, 97.6%, 99.8% and 100% in the sarolaner-treated group and 94.1%, 88.5%, 89.9% and 98.1% in the fipronil-treated group on Days 14, 30, 60 and 90, respectively. Sarolaner was non-inferior to fipronil at all time-points, and was superior on Days 30 and 60. Sarolaner tablets were voluntarily and fully consumed within one minute in 93% of the 1280 occasions offered.Sarolaner administered orally at monthly intervals at a minimum dosage of 2 mg/kg was safe and highly effective against natural infestations of fleas and ticks on dogs. In addition, clinical signs FAD improved in dogs treated with sarolaner, and the flavored, chewable tablets were highly palatable.

The efficacy of the novel isoxazoline, sarolaner (Simparica™) was investigated in dogs with clinical signs consistent with sarcoptic mange and harbouring natural infestations of Sarcoptes scabiei. One placebo-controlled laboratory study and one multi-centred field study with a commercial comparator containing imidacloprid/moxidectin (Advocate® spot-on) were conducted. Oral or topical treatments were administered on Days 0 and 30. Up to 10 skin scrapings were taken for the assessment of S. scabiei infestations from each dog before treatment and on Days 14, 30, 44 and 60 in the laboratory study, and on Days 30 and 60 in the field study. In the laboratory study, efficacy was calculated based on the percent reduction of mean live mite counts compared to the placebo group. In the field study parasitological cure rate (% dogs free of mites) was determined and non-inferiority of sarolaner to the control product was assessed.In the laboratory study 44 mixed breed dogs were enrolled in four batches. Due to decreasing mite counts in the placebo treated dogs, immunosuppression with dexamethasone (0.4mg/kg three times per week for two weeks) was initiated in all dogs on study at that time (n=6) and those subsequently enrolled (n=14). In the field study, dogs were enrolled in a 2:1 ratio (sarolaner:comparator); 79 dogs were assessed for efficacy and safety, and an additional 45 dogs were assessed for safety only. There were no treatment related adverse events in either study.In the laboratory study, no mites were found on any sarolaner-treated dogs 14 days after the first treatment except for one dog that had a single mite on Day 44. In the field study, the parasitological cure rate was 88.7% and 100% in the sarolaner group and 84.6% and 96.0% in the imidacloprid/moxidectin group, on Days 30 and 60, respectively. Statistical analysis showed that sarolaner was non-inferior to imidacloprid/moxidectin at both time points. The clinical signs of sarcoptic mange, including hair loss, papules, pruritus, erythema, and scaling/crusting improved throughout the study.Sarolaner was safe, achieved 100% reduction in the numbers of S. scabiei detected and resulted in marked improvement of the clinical signs of sarcoptic mange in dogs following two monthly oral administrations.

The efficacy of a single oral dose of a novel isoxazoline, sarolaner (Simparica™, Zoetis), for the treatment and control of flea infestations on dogs was confirmed in five laboratory studies. The studies were conducted using adult purpose-bred Beagles and/or mixed breed dogs. All animals were individually identified and housed, and were allocated randomly to treatment with either placebo or sarolaner (eight to 10 per group) based on pretreatment parasite counts. Three studies used cat flea (Ctenocephalides felis felis) strains recently isolated from the field from the US, EU, or Australia; in the fourth study a laboratory strain (KS1) with documented tolerance to a number of insecticides such as fipronil, imidacloprid, and permethrin was used. In the fifth study, dogs were infested with dog fleas, Ctenocephalides canis. Dogs were treated orally on Day 0 with a placebo or a sarolaner tablet providing a minimum dose of 2mg/kg. Dogs were infested with approximately 100 unfed, adult fleas prior to treatment and at weekly intervals post-treatment. Comb counts were conducted to determine the numbers of viable fleas at 24h after treatment and after each subsequent infestation. Efficacy against C. felis and C. canis was 99.8–100% from treatment through Day 35. In all five studies, elimination of existing infestations was achieved within 24h after dosing, with only a single live C. felis found on one dog on Day 1. Similarly, control of flea challenges was achieved within 24h after infestation throughout the 35day study periods, with only single live C. felis found on two dogs on Day 28 in one study, and on a single dog on Day 35 in another study. There were no adverse reactions to treatment with sarolaner. These studies confirmed that a single oral dose of sarolaner at 2mg/kg provided highly effective treatment of existing C. felis infestations and persistent control of C. felis on dogs for 35days after treatment. Efficacy equivalent to that seen with C. felis was confirmed against C. canis and a known insecticide-tolerant strain of C. felis.

Lipid A on the Gram-negative outer membrane (OM) is synthesized in the cytoplasm by the Lpx pathway and translocated to the OM by the Lpt pathway. Some Acinetobacter baumannii strains can tolerate the complete loss of lipopolysaccharide (LPS) resulting from the inactivation of early LPS pathway genes such as lpxC . Here, we characterized a mutant deleted for lptD , which encodes an OM protein that mediates the final translocation of fully synthesized LPS to the OM. Cells lacking lptD had a growth defect comparable to that of an lpxC deletion mutant under the growth conditions tested but were more sensitive to hydrophobic antibiotics, revealing a more significant impact on cell permeability from impaired LPS translocation than from the loss of LPS synthesis. Consistent with this, ATP leakage and N -phenyl-1-naphthylamine (NPN) fluorescence assays indicated a more severe impact of lptD deletion than of lpxC deletion on inner and outer membrane permeability, respectively. Targeted liquid chromatography-mass spectrometry (LCMS) analysis of LPS intermediates from UDP-3- O-R -3-hydroxylauroyl- N -acetyl-α- d -glucosamine through lipid IV A showed that the loss of LptD caused an accumulation of lipid IV A . This suggested that pathway intermediate accumulation or mislocalization caused by the blockage of later LPS pathway steps impacts envelope integrity. Supporting this notion, chemical inhibition of lipid A precursor enzymes, including LpxC and FabB/F, in the lptD deletion strain partially rescued growth and permeability defects. IMPORTANCE New antibiotics to treat Gram-negative bacterial infections are urgently needed. Inhibition of LPS biosynthesis is attractive because this would impact viability and cell permeability. Therefore, a better understanding of this pathway is important, especially in strains such as A. baumannii ATCC 19606, where LPS biosynthesis is not essential in vitro . We show that ATCC 19606 also survives the loss of the final translocation of LPS into the OM ( lptD deletion). Intriguingly, this impaired cell envelope integrity more than the loss of LPS biosynthesis ( lpxC deletion), presumably due to the accumulation of toxic intermediates. Supporting this, chemical inhibition of LPS biosynthesis partially reversed this permeability defect. This extends our understanding of the LPS machinery and provides insights into potential interrelationships of the target steps along this important pathway.

Purpose Treatments that delay retinal cell death regardless of genetic causation are needed for inherited retinal degeneration (IRD) patients. The ketogenic diet is a high-fat, low-carbohydrate diet, used to treat epilepsy, and has beneficial effects for neurodegenerative diseases. This study aimed to determine whether the ketogenic diet could slow retinal degeneration. Methods Early weaned, rd10 and wild-type (WT) mice were placed on either standard chow, a ketogenic diet, or a ketogenic & low-protein diet. From postnatal day (PD) 23 to PD50, weight and blood β-hydroxybutyrate levels were recorded. Retinal thickness, retinal function, and visual performance were measured via optical coherence tomography, electroretinography (ERG), and optokinetic tracking (OKT). At PD40, serum albumin, rhodopsin protein, and phototransduction gene expression were measured. Results Both ketogenic diets elicited a systemic induction of ketosis. However, rd10 mice on the ketogenic & low-protein diet had significant increases in photoreceptor thickness, ERG amplitudes, and OKT thresholds, whereas rd10 mice on the ketogenic diet showed no photoreceptor preservation. In both rd10 and WT mice, the ketogenic & low-protein diet was associated with abnormal weight gain and decreases in serum albumin levels, 27% and 56%, respectively. In WT mice, the ketogenic & low-protein diet was also associated with an ∼20% to 30% reduction in ERG amplitudes. Conclusions The ketogenic & low-protein diet slowed retinal degeneration in a clinically relevant IRD model. In WT mice, the ketogenic & low-protein diet was associated with a decrease in phototransduction and serum albumin, which could serve as a protective mechanism in the rd10 model. Although ketosis was associated with protection, its role remains unclear. Translational relevance Neuroprotective mechanisms associated with the ketogenic & low-protein diet have potential to slow retinal degeneration.