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Pseudomonas aeruginosa, a major cause of nosocomial infections, is considered a paradigm of antimicrobial resistance, largely due to hyperproduction of chromosomal cephalosporinase AmpC. Here, we explore the ability of 6-pyridylmethylidene penicillin-based sulfones 1-3 to inactivate the AmpC β-lactamase and thus rescue the activity of the antipseudomonal ceftazidime. These compounds increased the susceptibility to ceftazidime in a collection of clinical isolates and PAO1 mutant strains with different ampC expression levels and also improved the inhibition kinetics relative to avibactam, displaying a slow deacylation rate and involving the formation of an indolizine adduct. Bromide 2 was the inhibitor with the lowest KI (15.6 nM) and the highest inhibitory efficiency (kinact/KI). Computational studies using diverse AmpC enzymes revealed that the aromatic moiety in 1-3 targets a tunnel-like site adjacent to the catalytic serine and induces the folding of the H10 helix, indicating the potential value of this not-always-evident pocket in drug design.

Zinc, cadmium, and lead in livers, kidneys, and feathers of 48 young and adult Atlantic puffins found dead or dying off the coast of Galicia (Northwest Spain) were determined. The most abundant bet...

BackgroundPseudomonas aeruginosa may develop resistance to novel cephalosporin/β-lactamase inhibitor combinations during therapy through the acquisition of structural mutations in AmpC.ObjectivesTo describe the molecular and biochemical mechanisms involved in the development of resistance to ceftolozane/tazobactam in vivo through the selection and overproduction of a novel AmpC variant, designated PDC-315.MethodsPaired susceptible/resistant isolates obtained before and during ceftolozane/tazobactam treatment were evaluated. MICs were determined by broth microdilution. Mutational changes were investigated through WGS. Characterization of the novel PDC-315 variant was performed through genotypic and biochemical studies. The effects at the molecular level of the Asp245Asn change were analysed by molecular dynamics simulations using Amber.ResultsWGS identified mutations leading to modification (Asp245Asn) and overproduction of AmpC. Susceptibility testing revealed that PAOΔC producing PDC-315 displayed increased MICs of ceftolozane/tazobactam, decreased MICs of piperacillin/tazobactam and imipenem and similar susceptibility to ceftazidime/avibactam compared with WT PDCs. The catalytic efficiency of PDC-315 for ceftolozane was 10-fold higher in relation to the WT PDCs, but 3.5- and 5-fold lower for piperacillin and imipenem. IC50 values indicated strong inhibition of PDC-315 by avibactam, but resistance to cloxacillin inhibition. Analysis at the atomic level explained that the particular behaviour of PDC-315 is linked to conformational changes in the H10 helix that favour the approximation of key catalytic residues to the active site.ConclusionsWe deciphered the precise mechanisms that led to the in vivo emergence of resistance to ceftolozane/tazobactam in P. aeruginosa through the selection of the novel PDC-315 enzyme. The characterization of this new variant expands our knowledge about AmpC-mediated resistance to cephalosporin/β-lactamase inhibitors in P. aeruginosa.

The global distribution of carbapenemases such as KPC, OXA-48, and metallo-β-lactamases (MBLs) gives cause for concern, as these enzymes are not inhibited by classical β-lactamase inhibitors (BLIs). The current development of new inhibitors is one of the most promising highlights for the treatment of multidrug-resistant bacteria. The activity of cefepime in combination with the novel BLIs zidebactam, taniborbactam, and enmetazobactam was studied in a collection of 400 carbapenemase-producing Enterobacterales (CPE). The genomes were fully sequenced and potential mechanisms of resistance to cefepime/BLI combinations were characterized. Cefepime resistance in the whole set of isolates was 79.5% (MIC50/90 64/≥128mg/L). The cefepime/zidebactam and cefepime/taniborbactam combinations showed the highest activity (MIC(50/90) ≤0.5/1 and ≤0.5/2 mg/L, respectively). Cefepime/zidebactam displayed high activity, regardless of the carbapenemase or extended-spectrum β-lactamase (ESBL) considered (99% of isolates displayed MIC ≤2 mg/L). Cefepime/taniborbactam displayed excellent activity against OXA-48- and KPC-producing Enterobacterales and lower activity against MBL-producing isolates (four strains yielded MICs ≥16 mg/L: 2 NDM producers with an insertion in PBP3, one VIM-1 producer with nonfunctional OmpK35, and one IMP-8 producer). Cefepime/enmetazobactam displayed the lowest activity (MIC(50/90) 1/≥128 mg/L), with MICs ≥16 mg/L for 49 MBL producers, 40 OXA-48 producers (13 with amino acid changes in OmpK35/36, 4 in PBPs and 11 in RamR) and 25 KPC producers (most with an insertion in OmpK36). These results confirm the therapeutic potential of the new β-lactamase inhibitors, shedding light on the activity of cefepime and BLIs against CPE and resistance mechanisms. The cefepime/zidebactam and cefepime/taniborbactam combinations are particularly highlighted as promising alternatives to penicillin-based inhibitors for the treatment of CPE.

The value of birds as bioindicators for monitoring the environmental inorganic elements has been globally recognized. In this context, due to its well-known ecology and population stability, the Northern gannet (Morus bassanus) could be particularly useful. Dead Northern gannets (n = 30) were collected and samples from the liver, kidney, and feathers were taken, dried, mineralized, and finally analyzed via ICP-MS. Metals and metalloids, namely As, Cd, Hg, Pb, and Zn, associated with environmental pollution and toxicity on living organisms, were evaluated. The mean highest concentrations of As, Hg, and Zn were found in the liver (0.916, 7.026, and 89.81 mg/kg dry weight, respectively). For Cd, the kidney showed the highest mean concentration (17.51 mg/kg dry weight), whereas for Pb, this value corresponded to the feathers (0.399 mg/kg dry weight). Significant differences were found between the age classes in terms of contaminant concentrations, with the adults exhibiting higher metal levels. This difference was significantly relevant for Pb and Hg, where the effect of age was observed for all the considered tissues. When considering the effect of gender, no significant differences were observed, in agreement with similar studies performed in other geographical regions. Finally, positive correlations between the concentrations of Hg and Pb in the feathers and in the liver (r = 0.688, p < 0.001 and r = 0.566, p < 0.001, respectively) were observed, as well as between the feather and kidney concentrations (r = 0.685, p < 0.001) indicating the possibility to use feathers, a non-invasive biomonitoring tissue, for better understanding Hg and Pb exposure in seabirds.

[Abstract] A kinetic model is proposed to fit isothermal thermogravimetric data obtained from cellulose in an inert atmosphere at different temperatures. The method used here to evaluate the model involves two steps: (1) fitting of single time-derivative thermogravimetric curves (DTG) obtained at different temperatures versus time, and (2) fitting of the rate parameter values obtained at different temperatures versus temperature. The first step makes use of derivative of logistic functions. For the second step, the dependence of the rate factor on temperature is evaluated. That separation of the curve fitting from the analysis of the rate factor resulted to be very flexible since it proved to work for previous crystallization studies and now for thermal degradation of cellulose Ministerio de Asuntos Económicos y Transformación Digital; MTM2017-82724-R Xunta de Galicia; ED431C-2020-14 Xunta de Galicia; ED431G 2019/01

Carbapenem resistance is a major global health problem that seriously compromises the treatment of infections caused by nosocomial pathogens. Resistance to carbapenems mainly occurs via the production of carbapenemases, such as VIM, IMP, NDM, KPC and OXA, among others. Preclinical and clinical trials are currently underway to test a new generation of promising inhibitors, together with the recently approved avibactam, relebactam and vaborbactam. This review summarizes the main, most promising carbapenemase inhibitors synthesized to date, as well as their spectrum of activity and current stage of development. We particularly focus on β-lactam/β-lactamase inhibitor combinations that could potentially be used to treat infections caused by carbapenemase-producer pathogens of critical priority. The emergence of these new combinations represents a step forward in the fight against antimicrobial resistance, especially in regard to metallo-β-lactamases and carbapenem-hydrolysing class D β-lactamases, not currently inhibited by any clinically approved inhibitor.

The emergence of 16S rRNA methyltransferases (RMTs) in gram-negative pathogens bearing other clinically relevant resistance mechanisms, such as carbapenemase-producing Enterobacterales (CPE), is becoming an alarming concern. We investigated the prevalence, antimicrobial susceptibility, resistance mechanisms, molecular epidemiology, and genetic support of RMTs in CPE isolates from Spain. The study included a collection of 468 CPE isolates recovered during 2018 from 32 participating Spanish hospitals. MICs were determined using the broth microdilution method, the agar dilution method (fosfomycin) or MIC gradient strips (plazomicin). All isolates were subjected to hybrid whole-genome sequencing. Sequence types (STs), core-genome phylogenetic relatedness, horizontally acquired resistance mechanisms, plasmid analysis and genetic environment of RMTs was in silico determined from WGS data in all RMT-positive isolates. Among the 468 CPE isolates evaluated, 24 (5.13%) isolates recovered from 9 different hospitals spanning 5 different Spanish regions showed resistance to all aminoglycosides and were positive for an RMT: 21 RmtF, 2 AmrA and 1 RmtC. All the RMT-producers showed high-level resistance to all aminoglycosides, including plazomicin, and in most of cases exhibited an extensively drug-resistant (XDR) susceptibility profile. The RMT-positive isolates showed low genetic diversity and were global clones of K. pneumoniae (ST147, ST101, ST395) or E. cloacae (ST93) species bearing blaOXA-48, blaNDM-1 or blaVIM-1 carbapenemase genes. RMTs were in 5 different multidrug-resistant plasmids and linked to efficient mobile elements. Our findings highlight that RMTs are emerging among clinical CPE isolates from Spain and their spread should be monitored to preserve the clinical utility of aminoglycosides and plazomicin in the future.

Background Imipenem/relebactam is a novel carbapenem/β-lactamase inhibitor combination, developed to act against carbapenemase-producing Enterobacterales (CPE). Objectives To assess the in vitro activity of imipenem/relebactam against a Spanish nationwide collection of CPE by testing the susceptibility of these isolates to 16 widely used antimicrobials and to determine the underlying β-lactam resistance mechanisms involved and the molecular epidemiology of carbapenemases in Spain. Materials and methods Clinical CPE isolates (n = 401) collected for 2 months from 24 hospitals in Spain were tested. MIC50, MIC90 and susceptibility/resistance rates were interpreted in accordance with the EUCAST guidelines. β-Lactam resistance mechanisms and molecular epidemiology were characterized by WGS. Results For all isolates, high rates of susceptibility to colistin (86.5%; MIC50/90 = 0.12/8 mg/L), imipenem/relebactam (85.8%; MIC50/90 = 0.5/4 mg/L) and ceftazidime/avibactam (83.8%, MIC50/90 = 1/≥256 mg/L) were observed. The subgroups of isolates producing OXA-48-like (n = 305, 75.1%) and KPC-like enzymes (n = 44, 10.8%) were highly susceptible to ceftazidime/avibactam (97.7%, MIC50/90 = 1/2 mg/L) and imipenem/relebactam (100.0%, MIC50/90 = ≤0.25/1 mg/L), respectively. The most widely disseminated high-risk clones of carbapenemase-producing Klebsiella pneumoniae across Spain were found to be ST11, ST147, ST392 and ST15 (mostly associated with OXA-48) and ST258/512 (in all cases producing KPC). Conclusions Imipenem/relebactam, colistin and ceftazidime/avibactam were the most active antimicrobials against all CPEs. Imipenem/relebactam is a valuable addition to the antimicrobial arsenal used in the fight against CPE, particularly against KPC-producing isolates, which in all cases were susceptible to this combination.

The ability of 6-(aryl)methylidene penicillin-based sulfones 1-7 to repurpose β-lactam antibiotics activity with bacterial species that carry carbapenem-hydrolyzing class D β-lactamases (OXA-23, OXA-24/40 and OXA-48), as well as with class A (TEM-1, CTX-M-2) and class C (CMY-2, DHA-1) enzymes, is reported. The combinations imipenem/3 and imipenem/4 restored almost completely the antibiotic efficacy in OXA-23 and OXA-24/40 carbapenemase-producing A. baumannii strains (1 μg mL-1) and also provided good results for OXA-48 carbapenemase-producing K. pneumoniae strains (4 μg mL-1). Compounds 2-6 in combinations with ceftazidime and ampicillin were also efficient in restoring antibiotic efficacy in E. coli strains carrying class C (CMY-2 and DHA-1) and class A (TEM-1 and CTX-M-2) β-lactamase enzymes, respectively. Kinetic and inhibition studies with the OXA-24/40 enzyme, protein mass spectrometry analysis and docking studies allowed us to gain an insight into the inhibition mechanism and the experimentally observed differences between the ligands.

We have prepared two new lead halides with the novel general formula of DMA7Pb4X15 (DMA = [(CH3)2NH2]+ and X = Cl– or Br–) by using an easy route under mild conditions at room temperature. These compounds exhibit an unprecedented crystal structure, are formed by layers of distorted [PbX6] octahedra, which share corners and faces, and contain intercalated DMA cations. Very interestingly, they display dielectric transitions, which are related to a partial order–disorder process of the DMA cations between 160 and 260 K. Additionally, these new layered hybrids exhibit a broadband photoluminiscent emission, which is related to the structural distortions of the [PbX6] octahedra. These findings not only open up large possibilities for future optoelectronic applications of these materials, but they also offer a novel playground for an easy modulation of electrical and optical properties of hybrid organic–inorganic materials. We anticipate that this novel A7Pb4X15 formula can be adequate to tune the family of the ...

This article belongs to the Special Issue Thermal and Rheological Characterization of Polymeric Materials [Abstract] Poly(lactic acid) (PLA) is an attractive candidate for replacing petrochemical polymers because it is fully biodegradable. This study investigated the potential of PLA as a sustainable and environmentally friendly alternative material that can be developed into commercially viable wearable mosquito repellent devices with desirable characteristics. PLA strands containing DEET and IR3535 were prepared by twin screw extrusion compounding and simultaneously functioned as plasticizers for the polymer. The plasticizing effect was investigated by thermal and rheological studies. DSC studies showed that the addition of DEET and IR3535 into PLA strands reduced the glass transition temperature consistent with predictions of the Fox equation, thus proving their efficiency as plasticizers. The rheology of molten samples of neat PLA and PLA/repellents blends, evaluated at 200 °C, was consistent with shear-thinning pseudoplastic behaviour. Raman studies revealed a nonlinear concentration gradient for DEET in the PLA strand, indicating non-Fickian Type II transport controlling the desorption process. Release data obtained at 50 °C showed initial rapid release followed by a slower, near constant rate at longer times. The release rate data were fitted to a novel modification of the Peppas-Sahlin desorption model This work was financially supported from the Deutsche Forschungsgemeinschaft (DFG), Germany [Grant number AN 212/22-2] and University of Pretoria (UP) Postdoctoral Fellowship programme. The research has been also supported by Spanish MINECO grant MTM2017-82724-R through the ERDF Deutsche Forschungsgemeinschaft (DFG); AN 212/22-2 https://www.mdpi.com/journal/molecules/special_issues/Thermal_Rheological_Polymeric_Materials

Background. Data on durability of first-line antiretroviral therapy (ART) in children with human immunodeficiency virus (HIV) are limited. We assessed time to switch to second-line therapy in 16 European countries and Thailand.Methods. Children aged = 2 nucleoside reverse transcriptase inhibitors p[NRTIs] plus nonnucleoside reverse transcriptase inhibitor p[NNRTI] or boosted protease inhibitor p[PI]) were included. Switch to second-line was defined as (i) change across drug class (PI to NNRTI or vice versa) or within PI class plus change of >= 1 NRTI; (ii) change from single to dual PI; or (iii) addition of a new drug class. Cumulative incidence of switch was calculated with death and loss to follow-up as competing risks.Results. Of 3668 children included, median age at ART initiation was 6.1 (interquartile range (IQR), 1.7-10.5) years. Initial regimens were 32% PI based, 34% nevirapine (NVP) based, and 33% efavirenz based. Median duration of follow-up was 5.4 (IQR, 2.9-8.3) years. Cumulative incidence of switch at 5 years was 21% (95% confidence interval, 20%-23%), with significant regional variations. Median time to switch was 30 (IQR, 16-58) months; two-thirds of switches were related to treatment failure. In multivariable analysis, older age, severe immunosuppression and higher viral load (VL) at ART start, and NVP-based initial regimens were associated with increased risk of switch.Conclusions. One in 5 children switched to a second-line regimen by 5 years of ART, with two-thirds failure related. Advanced HIV, older age, and NVP-based regimens were associated with increased risk of switch.

A method based on Matrix Solid Phase Dispersion (MSPD) for the determination of nine triazines in marine sediments was optimized in terms of dispersant agents, co-sorbents and elution solvents. Three dispersing agents: octadecylsilyl bonded silica (C18), graphitized carbon black (GCB) and diatomaceous earth, and four clean-up co-sorbents: Florisil, GCB, primary and secondary amine (PSA)/strong anion exchanger (SAX), and GCB/PSA were assayed. The best results were obtained with GCB as dispersing agent, without co-sorbent, using 20 mL of ethyl acetate as elution solvent. Finally the extract was concentrated to dryness, re-constituted with 1 mL methanol and determined by HPLC-DAD. The analytical recoveries obtained were close to 100% and repeatability and reproducibility were below than 3.5% for all compounds. The linearity of the calibration curves was excellent in matrix matched standards, and yielded the coefficients of determination ≥ 0.9992 for all the target analytes. The LOQ values ranged from 0.022 to 0.037 mg kg − 1 . Finally the method was applied to analyse the target compounds in marine sediment samples from the estuary of A Coruna (Galicia, NW of Spain).

The fast growing family of organic–inorganic hybrid compounds has recently been attracting increased attention owing to the remarkable functional properties (magnetic, multiferroic, optoelectronic, photovoltaic) displayed by some of its members. Here we show that these compounds can also have great potential in the until now unexplored field of solid-state cooling by presenting giant barocaloric effects near room temperature already under easily accessible pressures in the hybrid perovskite [TPrA][Mn(dca)3] (TPrA: tetrapropylammonium, dca: dicyanamide). Moreover, we propose that this will not be an isolated example for such an extraordinary behaviour as many other organic–inorganic hybrids (metal-organic frameworks and coordination polymers) exhibit the basic ingredients to display large caloric effects which can be very sensitive to pressure and other external stimuli. These findings open up new horizons and great opportunities for both organic–inorganic hybrids and for solid-state cooling technologies., Designing mechanically and chemically robust liquid-repellent surfaces remains a long-standing challenge. Here, Wang and colleagues report a microfluidic emulsion templating strategy to fabricate bioinspired omniphobic porous membranes with remarkable durability.

In this work, we focus on [(CH3)2NH2]PbI3, a member of the [AmineH]PbI3 series of hybrid organic–inorganic compounds, reporting a very easy mechanosynthesis route for its preparation at room temperature. We report that this [(CH3)2NH2]PbI3 compound with 2H-perovskite structure experiences a first-order transition at ≈250 K from hexagonal symmetry P63/mmc (HT phase) to monoclinic symmetry P21/c (LT phase), which involves two cooperative processes: an off-center shift of the Pb2+ cations and an order–disorder process of the N atoms of the DMA cations. Very interestingly, this compound shows a dielectric anomaly associated with the structural phase transition. Additionally, this compound displays very large values of the dielectric constant at room temperature because of the appearance of a certain conductivity and the activation of extrinsic contributions, as demonstrated by impedance spectroscopy. The large optical band gap displayed by this material (Eg = 2.59 eV) rules out the possibility that the observ...

The pyrrole-imidazoles, a group of alkaloids commonly found in marine sponges belonging to the genus Agelas, display a wide range of biological activities. Herein, we report the first chemical study of the secondary metabolites of the sponge A. dilatata from the coastal area of the Yucatan Peninsula (Mexico). In this study, we isolated eight known alkaloids from an organic extract of the sponge. We used NMR and MS analysis and comparison with existing databases to characterize the alkaloids: ageliferin (1), bromoageliferin (2), dibromoageliferin (3), sceptrin (4), nakamuric acid (5), 4-bromo-1H-pyrrole-2-carboxylic acid (6), 4,5-dibromopyrrole-2-carboxylic acid (7) and 3,7-dimethylisoguanine (8). We also evaluated, for the first time, the activity of these alkaloids against the most problematic multidrug-resistant (MDR) pathogens, i.e., the Gram-negative bacteria Pseudomonas aeruginosa, Klebsiella pneumoniae and Acinetobacter baumannii. Bromoageliferin (2) displayed significant activity against P. aeruginosa. Comparison of the antibacterial activity of ageliferins 1&ndash, 3 (of similar structure) against P. aeruginosa revealed some relationship between structure and activity. Furthermore, in in vitro assays, 2 inhibited growth and biofilm production in clinical strains of P. aeruginosa. Moreover, 2 increased the survival time in an in vivo Galleria mellonella model of infection. The findings confirm bromoageliferin (2) as a potential lead for designing new antibacterial drugs.

The carbapenem-hydrolyzing class D β-lactamases (CHDLs) are the main mechanism of carbapenem resistance in Acinetobacter baumannii. CHDLs are not effectively inactivated by clinically available β-lactam-type inhibitors. We have previously described the in vitro efficacy of the inhibitor LN-1-255 in combination with carbapenems. The aim of this study was to compare the efficacy of LN-1-255 with that of imipenem in murine pneumonia using A. baumannii strains carrying their most extended carbapenemases, OXA-23 and OXA-24/40. The bla(OXA-23) and bla(OXA-24/40) genes were cloned into the carbapenem-susceptible A. baumannii ATCC 17978 strain. Clinical isolates Ab1 and JC12/04, producing the enzymes OXA-23 and OXA-24/40, respectively, were used in the study. Pharmacokinetic (PK) parameters were determined. An experimental pneumonia model was used to evaluate the efficacy of the combined imipenem–LN-1-255 therapy. MICs of imipenem decreased between 32- and 128-fold in the presence of LN-1-255. Intramuscular treatment with imipenem–LN-1-255 (30/50 mg/kg) decreased the bacterial burden by (i) 4 and 1.7 log(10) CFU/g lung in the infection with the ATCC 17978-OXA-23 and Ab1 strains, respectively, and by (ii) 2.5 and 4.5 log(10) CFU/g lung in the infection produced by the ATCC 17978-OXA-24/40 and the JC12/04 strains, respectively. In all assays, combined therapy offered higher protection against pneumonia than that provided by monotherapy. No toxicity was observed in treated mice. Imipenem treatment combined with LN-1-255 treatment significantly reduced the severity of infection by carbapenem-resistant A. baumannii strains carrying CHDLs. Preclinical assays demonstrated the potential of LN-1-255 and imipenem therapy as a new antibacterial treatment.

Concentrations of 7 different polychlorinated biphenyl (PCB) congeners, and eleven organochlorine pesticides (OCPs) and metabolites, including DDTs (dichlorodiphenyltrichloroethane), HCHs (hexachlorocyclohexane isomers), Endosulfan, Endosulfan sulfate, Endrin, Dieldrin and HCB (hexachlorobenzene), were determined in adipose tissue of 57 yellow-legged gulls collected from NW and N Spain. Furthermore, the possible differences due to two endogenous factors, age and gender, were determined. All the analyzed PCBs were detected in over 66% of the samples, with levels of 291.9 (PCB 180), 34.5 (PCB 118), 0.7 (PCB 28), 432.6 (PCB 153), 225.5 (PCB 138), 1.3 (PCB 101) and 0.4 (PCB 52) µg/kg of adipose tissue. With respect to the OCPs and metabolites, only 4,4′-DDE and HCB were detected in more than 50% of the samples, with means of 360.6 and 2.5 µg/kg of adipose tissue, respectively. From all the considered contaminants, only 4,4′-DDE levels presented significant differences depending on the gender, with females showing higher values than males (p < 0.01). Significant differences (p < 0.001) were also found related to age for the levels of PCBs 180, 138, 101, 28 and 153, as well as 4,4′-DDE, with adult levels being higher than those in young birds. The results of the present study constitute a baseline to better assess the environmental impacts of PCB and OCP contamination at other coastal sites for future biomonitoring studies, with particular emphasis on gender- and age-related differences.

A fast, simple, selective and sensitive method has been developed for the determination of nine triazine herbicides (ametryn, atrazine, cyanazine, prometryn, propazine, simazine, simetryn, terbuthylazine and terbutryn) and eight degradation products (desethyl atrazine, desethyl-desisopropyl atrazine, desethyl 2-hydroxyatrazine, desethyl terbuthylazine, desisopropyl atrazine, desisopropyl 2-hydroxyatrazine, 2-hydroxyatrazine and 2-hidroxyterbuthylazine) in seawater samples. On-line solid-phase extraction coupled with ultra-pressure liquid chromatography-tandem mass spectrometry was employed for simultaneous analysis of all compounds in 11min. Validation parameters were studied through the estimation of the limits of detection and quantification, calibration curves and precision. Limits of quantification ranged from 0.023 to 0.657μgL-1. Good linearity was obtained for all compounds with R2>0.99 in all cases. Furthermore, inter-day precision (0-2.1%) and intra-day precision (0-3.9%) were shown to be satisfactory. On-line solid-phase extraction recoveries in spiked unpolluted seawater sample were evaluated and acceptable values (80.3-99.8%) with adequate RSD (0.1-3.1%) were found. Finally, the proposed method was applied to the analysis of the target compounds in seawater samples collected from seawater nearby a zone of intensive horticulture of Matosinhos (Portugal). The concentrations of the herbicides were below the limit of detection in all cases.

In this work we prepare and study the M(dca) 2 pyz compounds, where dca = polar dicyanamide N(CN) 2 − ligand, pyz = pyrazine (C 4 H 4 N 2 ) ligand and M = Fe 2+ , Co 2+ and Zn 2+ that, according to DSC display a first order phase transition, at T t ∼ 270 K (Fe), ∼250 K (Co) and ∼240 K (Zn). Very interestingly these compounds, which consist of a rather robust framework with incorporated polar dca ligands, and which does not host guest species in the cavities of the structure, display a dielectric anomaly associated to such phase transition. This latter in fact corresponds to the structural transition from the disordered HT-phase (space group Pnma ) to the ordered LT-phase (space group P 2 1 / n ). From a detailed analysis of the dipole distributions in each of these crystal structures we conclude that the polar dca ligands, which bridge two transition metal cations, display a cooperative electrical arrangement, in which the dipoles of each of the two interpenetrated networks show a ferroelectric (FE) arrangement; nevertheless, the relative antiparallel orientation of both networks finally results in net antiferroelectric (AFE) arrangements both in the LT- and HT-polymorphs. Very interestingly, the dynamic disorder of the dca ligands at T t gives rise to the observed dielectric anomaly. These materials constitute, therefore, an example of hybrid organic–inorganic compounds where the dielectric response directly arises from the framework –and not from polar guest species– which is a very interesting and novel approach in the search for materials in which the dielectric and magnetic properties can couple more strongly.

This is a pre-copyedited, author-produced version of an article accepted for publication in Journal of Antimicrobial Chemotherapy following peer review. The version of record Juan A. Vallejo, Marta Martínez-Guitián, Juan C. Vázquez-Ucha, Concepción González-Bello, Margarita Poza, John D. Buynak, Christopher R. Bethel, Robert A. Bonomo, German Bou, Alejandro Beceiro; LN-1-255, a penicillanic acid sulfone able to inhibit the class D carbapenemase OXA-48, Journal of Antimicrobial Chemotherapy, Volume 71, Issue 8, 1 August 2016, Pages 2171–2180 is available online at: https://doi.org/10.1093/jac/dkw105 Objectives Carbapenemases are the most important mechanism responsible for carbapenem resistance in Enterobacteriaceae. Among carbapenemases, OXA-48 presents unique challenges as it is resistant to β-lactam inhibitors. Here, we test the capacity of the compound LN-1-255, a 6-alkylidene-2′-substituted penicillanic acid sulfone, to inhibit the activity of the carbapenemase OXA-48. Methods The OXA-48 gene was cloned and expressed in Klebsiella pneumoniae and Escherichia coli in order to obtain MICs in the presence of inhibitors (clavulanic acid, tazobactam and sulbactam) and LN-1-255. OXA-48 was purified and steady-state kinetics was performed with LN-1-255 and tazobactam. The covalent binding mode of LN-1-255 with OXA-48 was studied by docking assays. Results Both OXA-48-producing clinical and transformant strains displayed increased susceptibility to carbapenem antibiotics in the presence of 4 mg/L LN-1-255 (2–32-fold increased susceptibility) and 16 mg/L LN-1-255 (4–64-fold increased susceptibility). Kinetic assays demonstrated that LN-1-255 is able to inhibit OXA-48 with an acylation efficiency (k2/K) of 10 ± 1 × 104 M−1 s−1 and a slow deacylation rate (koff) of 7 ± 1 × 10−4 s−1. IC50 was 3 nM for LN-1-255 and 1.5 μM for tazobactam. Lastly, kcat/kinact was 500-fold lower for LN-1-255 than for tazobactam. Conclusions In these studies, carbapenem antibiotics used in combination with LN-1-255 are effective against the carbapenemase OXA-48, an important emerging mechanism of antibiotic resistance. This provides an incentive for further investigations to maximize the efficacy of penicillin sulfone inhibition of class D plasmid-carried Enterobacteriaceae carbapenemases. This work was supported by the Spanish National Plans for Scientific Research, Development and Technological Innovation 2013-16 and funded by the ISCIII-General Subdirection of Assessment and Promotion of the Research-European Regional Development Fund (ERDF) ‘A way of making Europe’: PI12/00552 to G. B. and PI14/00059 to M. P. and A. B. Also, research reported in this publication was supported in part by the National Institute of Allergy and Infectious Diseases of the National Institutes of Health (USA) under Award Numbers R01AI100560, R01AI063517 and R01AI072219 to R. A. B. This study was supported in part by funds and/or facilities provided by the Cleveland Department of Veterans Affairs (USA), Award Number 1I01BX001974 to R. A. B. from the Biomedical Laboratory Research & Development Service of the VA Office of Research and Development and the Geriatric Research Education and Clinical Center VISN 10 (USA) to R. A. B. This study was also supported by the Spanish Ministry of Economy and Competiveness (SAF2013-42899-R), Xunta de Galicia (Spain) (GRC2013-041) and the European Regional Development Fund (ERDF) to C. G.-B, and supported by National Institutes of Health (USA) to J. D. B. (1R15AI109624). J. V. A. was financially supported by the Sara Borrell Programme ISCIII-FEDER (CD13/00373). J. V. H. and A. B. were financially supported by the Miguel Servet Programme ISCIII-FEDER (CP13/00226) SI

Acute myeloid leukemia (AML) can evade the mouse and human innate immune system by suppressing natural killer (NK) cell development and NK cell function. This is driven in part by the overexpression of microRNA (miR)-29b in the NK cells of AML patients, but how this occurs is unknown. In the current study, we demonstrate that the transcription factor aryl hydrocarbon receptor (AHR) directly regulates miR-29b expression. We show that human AML blasts activate the AHR pathway and induce miR-29b expression in NK cells, thereby impairing NK cell maturation and NK cell function, which can be reversed by treating NK cells with an AHR antagonist. Finally, we show that inhibition of constitutive AHR activation in AML blasts lowers their threshold for apoptosis and decreases their resistance to NK cell cytotoxicity. Together, these results identify the AHR pathway as a molecular mechanism by which AML impairs NK cell development and function. The results lay the groundwork in establishing AHR antagonists as potential therapeutic agents for clinical development in the treatment of AML.

Once pasteurized donor milk is thawed for its administration to a preterm or sick neonate, and until it is administered, it is kept refrigerated at 4 to 6°C for 24 h. After this time, unconsumed milk is discarded. This time has not been extended, primarily because of the concern of bacterial contamination. The aim of this study was to determine the changes in pH and bacterial count when pasteurized donor milk was kept under refrigeration for a prolonged period (14 d). In this prospective study, 30 samples of pasteurized donor milk from 18 donors were analyzed. Milk was handled following the regular operating protocols established in the neonatal unit and was kept refrigerated after thawing. pH measurements and bacteriology (on blood agar and MacConkey agar plates) were performed on each sample at time 0 (immediately after thawing) and then every day for 14 d. Changes in pH of samples over time were evaluated with linear mixed-effects regression models. A slow but gradual increase in milk pH was observed starting from the first day [mean (±SD) pH of 7.30 (±0.18) at time 0 and 7.69 (±0.2) on d 14]. No bacterial growth was observed in any of the samples throughout the complete trial except in one sample, in which Bacillus flexus was isolated. In conclusion, pasteurized human donor milk maintains its microbiological quality when properly handled and refrigerated (4-6°C). The slight and continuous increase in milk pH after the first day could be due to changes in the solubility of calcium and phosphate during refrigerated storage.

The hybrid inorganic–organic [TPrA][M(dca)3] (M: Fe2+, Co2+ and Ni2+) compounds, where TPrA is the tetrapropylammonium cation and dca is the dicyanamide anion, are unique multi-sensitive compounds that display multiple phases and dielectric transitions. These materials exhibit up to three first-order structural transitions (between the polymorphs I, Ia, Ib and II) associated with the same number of dielectric transitions in the temperature range of 210–360 K. The mechanisms responsible for these dielectric responses are found to be novel within the hybrid perovskites, involving ionic displacements of the A-site cations (TPrA) and order/disorder processes of the X anions (dca). In addition, the phase transitions and dielectric transition temperatures can be tuned by applying external hydrostatic pressure or by inducing internal pressure by modifying the tolerance factor through ionic substitution in the B-sites. This multi-sensitive response towards temperature, external and internal pressure opens up promising technological applications for this family of materials, such as dielectric transductors or multistimuli-sensors, whose response can be modulated in a wide range of temperatures and pressures., The authors are grateful for the financial support from Ministerio de Economía y Competitividad MINECO (MINECO) ENE2014-56237-C4-4-R and Xunta de Galicia under the project GRC2014/042. J. M. B.-G. also wants to thank Barrié Foundation for a predoctoral fellowship and S. Y.-V. to the Xunta de Galicia for a postdoctoral grant (Plan I2C).

Cd, Pb, and Zn were quantified in liver and kidney of red foxes (Vulpes vulpes) which were hunted during the 2003–2011 hunting seasons in Galicia (NW Spain). The effects of age and gender were evaluated to determine whether these variables should be included in future biomonitoring studies. The concentrations of hepatic and renal Cd (average 0.6 and 1.3 µg/g) and Pb (0.8 and 0.06 µg/g, respectively) were similar to background levels, with no known toxicological relevance. Similarly, the average levels of Zn in liver and kidney (77 and 17 µg/g) were in the range of physiological levels for canids. Although no significant gender-dependent variations were observed, the effect of aging was evident: the levels of hepatic Pb and both hepatic and renal Cd were higher in adults than in juveniles. Age should be included as a parameter during future biomonitoring programs focusing on trace metal bioaccumulation in red foxes.

The first example of an ammonium derivative that causes a specific modification of the active site of type I dehydroquinase (DHQ1), a dehydratase enzyme that is a promising target for antivirulence drug discovery, is described. The resolution at 1.35 Å of the crystal structure of DHQ1 from Salmonella typhi chemically modified by this ammonium derivative revealed that the ligand is covalently attached to the essential Lys170 through the formation of an amine. The detection by mass spectroscopy of the reaction intermediates, in conjunction with the results of molecular dynamics simulations, allowed us to explain the inhibition mechanism and the experimentally observed differences between S. typhi and Staphylococcus aureus enzymes. The results presented here reveal that the replacement of Phe225 in St-DHQ1 by Tyr214 in Sa-DHQ1 and its hydrogen bonding interaction with the conserved water molecule observed in several crystal structures protects the amino adduct against further dehydration/aromatization reactions. In contrast, for the St-DHQ1 enzyme, the carboxylate group of Asp114, with the assistance of this water molecule, would trigger the formation of a Schiff base that can undergo further dehydration reactions until full aromatization of the cyclohexane ring is achieved. Moreover, in vitro antivirulence studies showed that the reported compound is able to reduce the ability of Salmonella Enteritidis to kill A459 respiratory cells. These studies have identified a good scaffold for the design of irreversible inhibitors that can be used as drugs and has opened up new opportunities for the development of novel antivirulence agents by targeting the DHQ1 enzyme Financial support from the Spanish Ministry of Science and Innovation (SAF2013-42899-R), Xunta de Galicia (GRC2013-041), and the European Regional Development Fund (ERDF) is gratefully acknowledged. E.L. thanks the Xunta de Galicia for his postdoctoral fellowship. A.B. thanks the Miguel Servet Programme ISCIII-FEDER (CP13/00226) and the ISCIIIGeneral Subdirection of Assesment and Promotion of the Research (PI14/00059) for financial support SI

BACKGROUND Restoration of a balanced innate immune response is paramount to recovery from critical injury. Plasma transfusion may modulate innate immune responses; however, little is known about the immunomodulatory potential of various plasma products. We conducted in vitro experiments to determine the effects of fresh frozen plasma, thawed plasma, solvent/detergent plasma, and an investigational spray-dried solvent/detergent plasma product on monocyte function. METHODS Monocytes were isolated from healthy adult volunteers and cocultured with aliquots of autologous plasma (control), fresh frozen plasma, thawed plasma, solvent/detergent treated plasma, or spray-dried solvent/detergent plasma. Monocyte function was assessed by cytokine production with and without lipopolysaccharide (LPS) stimulation, and flow cytometric assessment of HLA-DR cell surface expression. RESULTS Monocyte cytokine production was not significantly altered after exposure to fresh frozen plasma or thawed plasma. In the absence of LPS, spray-dried solvent/detergent plasma exposure resulted in markedly increased IL-8 production compared to other plasma groups and controls (p = 0.01, analysis of variance [ANOVA]). Likewise, spray-dried SD plasma exposure resulted in higher LPS-induced IL-8, TNFα, and IL-1β production compared with autologous plasma controls (p < 0.0001; p < 0.0001, p = 0.002, respectively; ANOVA). LPS-induced IL-8 and TNFα production was lowest after exposure to solvent/detergent plasma (p < 0.0001, ANOVA). CONCLUSION Exposure to spray-dried solvent/detergent plasma resulted in marked augmentation of monocyte inflammatory cytokine production. Solvent/detergent plasma exposure resulted in the lowest cytokine production, suggesting lower immunomodulatory potential. Further work is needed to determine how these in vitro findings may translate to the bedside.

Quasifree one-proton knockout reactions have been employed in inverse kinematics for a systematic study of the structure of stable and exotic oxygen isotopes at the R3B/LAND setup with incident beam energies in the range of 300-450 MeV/u. The oxygen isotopic chain offers a large variation of separation energies that allows for a quantitative understanding of single-particle strength with changing isospin asymmetry. Quasifree knockout reactions provide a complementary approach to intermediate-energy one-nucleon removal reactions. Inclusive cross sections for quasifree knockout reactions of the type OA(p,2p)NA-1 have been determined and compared to calculations based on the eikonal reaction theory. The reduction factors for the single-particle strength with respect to the independent-particle model were obtained and compared to state-of-the-art ab initio predictions. The results do not show any significant dependence on proton-neutron asymmetry.

In view of a possible application as a charge-particle track readout for an Active-Target Time Projection Chamber (AT-TPC), the operational properties and performances of a hybrid Micro-Pattern Gaseous Detector (MPGD) were investigated in pure low-pressure Hydrogen (H2) and Helium (He). The detector consists of a MICROMEsh GAseous Structure (MICROMEGAS) coupled to a two-cascade THick Gaseous Electron Multiplier (THGEM) as a pre-amplification stage. This study reports the effective gain dependence of the hybrid-MPGD at relevant pressure (in the range of 200-760 torr) for different detector arrangements. The results of this work are relevant in the field of avalanche mechanism in low-pressure, low-mass noble gases, in particularly for applications of MPGD end-cap readout for active-target Time Projection Chambers (TPC) in the field of nuclear physics and nuclear astrophysics.

Acrylic copolymers and acrylates are of high interest in a wide variety of applications including coatings. This interest is increasing due to the possibility of being obtained by environmentally friendly procedures. In this research, the thermal stability in non-oxidizing atmosphere of two copolymers, a styrene/butyl acrylate and a diacetone acrylamide/butyl acrylate, is investigated by thermogravimetry (TG). A model consisting of a mixture of generalized logistic functions, which was used to fit calorimetric curves, was adapted to isothermal and non-isothermal contexts. The model was already applied to different materials and processes, being this time the first one that it is applied to TG degradation studies. In the current form, making use of multiple linear heating rates and isothermal experiments at several temperatures, the model allows for obtaining the true energy barrier and other kinetic parameters. The degradations of these copolymers were successfully fitted by the proposed model, and the main overlapping process was separately studied. The kinetic parameter values obtained from both compounds are compared to each other and to those reported from other cases where the model was applied. An important parameter is the critical temperature, which represents the minimum temperature for a given degradation processes to occur. Values of 495 and 525 K were obtained, respectively, for S/BA and BA/DAAM. True energy barrier values obtained for the degradation of these two polymers are approximately a half of those obtained in polymer crystallizations from the melt, and five times of those obtained in the case of an epoxy curing. The accelerating effect of applying a heating ramp is similar to that observed for polymer crystallization and smaller than that observed in thermoset curing.

The number of infections caused by Gram-negative pathogens carrying carbapenemases is increasing, and the group of carbapenem-hydrolyzing class D β-lactamases (CHDLs) is especially problematic. Several clinically important CHDLs have been identified in A. baumannii, including OXA-23, OXA-24/40, OXA-58, OXA-143, OXA-235, and the chromosomally encoded OXA-51. The selection and dissemination of carbapenem-resistant A. baumannii strains constitutes a serious global threat. Carbapenems have been successfully utilized as last resort antibiotics for the treatment of multi-drug-resistant A. baumannii infections. However, the spread of OXA carbapenemases is compromising the continued use of these antimicrobials. In response to this clinical issue, it is necessary and urgent to design and develop new specific inhibitors with efficacy against these enzymes. The aim of this work is to characterize the inhibitory activity of LN-1-255 (a 6-alkylidene-2-substituted penicillin sulfone) and compare it to that of two established inhibitors (avibactam and tazobactam) against the most relevant enzymes of each group of class D carbapenemases in A. baumannii. The β-lactamase inhibitor LN-1-255 demonstrated excellent microbiological synergy and inhibition kinetics parameters against all tested CHDLs, and a significantly higher activity than tazobactam and avibactam. A combination of carbapenems and LN-1-255 was effective against A. baumannii class D carbapenemases. Docking assays confirmed the affinity of LN-1-255 for the active site of these enzymes. LN-1-255 represents a potential new β-lactamase inhibitor, which may have a significant role in eradicating infections caused by A. baumannii isolates carrying CHDLs This work was supported by the Spanish National Plans for Scientific Research, Development and Technological Innovation 2008-2011 and 2013-2016 and funded by the ISCIII- General Subdirection of Assessment and Promotion of the Research-European Regional Development Fund (ERDF) “A way of making Europe”: PI12/00552 to G.B. and PI14/00059 to M.P. and A.B. Also, this study was supported in part by funds from the National Institute of Allergy and Infectious Diseases of the National Institutes of Health (USA) under award numbers R01AI063517 and R01AI100560, by funds and/or facilities provided by the Cleveland Department of Veterans Affairs, the Veterans Affairs Merit Review Program Award 1I01BX001974 and the Geriatric Research Education and Clinical Center VISN 10 to R.A.B., and by the Spanish Ministry of Economy and Competiveness (SAF2013-42899-R), Xunta de Galicia (GRC2013-041) and the European Regional Development Fund (ERDF) to C.GB. J.V. was financially supported by the Sara Borrell Programme ISCIII-FEDER (CD13/00373). J.V.H. and A.B. were financially supported by the Miguel Servet Programme ISCIII-FEDER (CP13/00226) SI

A fast, simple, sensitive and green chemistry method using dispersive liquid-liquid microextraction (DLLME) for the simultaneous determination of seven triazine herbicides (ametryn, atrazine, cyanazine, propazine, simazine, simetryn and terbuthylazine) in estuarine seawater samples has been developed. DLLME was carried out using a small volume of seawater (25 mL) and 300 μL of 1-octanol. Herbicide concentrations were determined by liquid chromatography-diode array detection, and results were confirmed by liquid chromatography-electrospray ionisation tandem spectrometry analysis. The analytical features of the proposed method were satisfactory with repeatability±5% and intermediate precision±10%, and recoveries ranged from 81-102% for all compounds. All the triazines exhibited linear matrix calibration curves with coefficients of determination0.999 for all the analytes except for simazine (0.9975). Limits of quantification ranged between 0.19 and 1.12 μg L(-1). The method was applied to the analysis of seawater samples from ten points susceptible to contamination by triazines from estuary of A Coruña (Galicia, NW of Spain). The levels of the seven triazines were below the LODs in the analysed samples. Use of proposed method will allow for monitoring of triazines at levels below the regulatory limits set by the European Directive 2008/105/EC of 2 and 4 μg L(-1) for atrazine and simazine, respectively.

A simple system consisting of a suspension of 10 wt % of starch in silicone oil was subjected to electrorheological testing. The system exhibits a complex behaviour depending on the electrical field. On one hand, the application of an external electric field induced the formation of a linear structure, aligned with the electric field. The formation of that structure was studied by several rheological methods. The rate of modifying the electric field intensity resulted to be related to the electric field value at which percolation is observed. On the other hand, master curves for the structure formation and breakdown in squeeze experiments were obtained.

A simple, effective, and economic method for determination of nine triazines (ametryn, atrazine, cyanazine, prometryn, propazine, simazine, simetryn, terbuthylazine, and terbutryn) in drinking water based on solid-phase extraction (SPE) followed by high-performance liquid chromatography-diode array detection (HPLC-DAD) was developed. A specialized solid phase (Oasis HLB) was used, and the parameters that may affect the efficiency of SPE were optimized. The limits of detection (ranged from 0.010 to 0.023 µg L−1) were satisfactory and allow the determination of triazines at the levels required by European Union legislation. Repeatability (2.4–7.6%) and intermediate precision (0.9–11.0%) calculated at 0.1 µg L−1 (legislation level) were adequate. The accuracy calculated as the average recovery of spiked tap and mineral waters was higher than 86% for all compounds. The developed method also could be used for undergraduate laboratory experiments because it acquaints students with solution preparation, solid ph...

Recent evidence suggests antimicrobial peptides protect the urinary tract from infection. Ribonuclease 7 (RNase 7), a member of the RNase A superfamily, is a potent epithelial-derived protein that maintains human urinary tract sterility. RNase 7 expression is restricted to primates, limiting evaluation of its antimicrobial activity in vivo . Here we identified ribonuclease 6 (RNase 6) as the RNase A superfamily member present in humans and mice that is most conserved at the amino acid level relative to RNase 7. Like RNase 7, recombinant human and murine RNase 6 has potent antimicrobial activity against uropathogens. Quantitative real-time PCR and immunoblot analysis indicate that RNase 6 mRNA and protein are upregulated in the human and murine urinary tract during infection. Immunostaining located RNase 6 to resident and infiltrating monocytes, macrophages, and neutrophils. Uropathogenic E. coli induces RNase 6 peptide expression in human CD14 + monocytes and murine bone marrow–derived macrophages. Thus, RNase 6 is an inducible, myeloid-derived protein with markedly different expression from the epithelial-derived RNase 7 but with equally potent antimicrobial activity. Our studies suggest RNase 6 serves as an evolutionarily conserved antimicrobial peptide that participates in the maintenance of urinary tract sterility.

Objectives Two natural variants of ADC-type β-lactamases of Acinetobacter spp., ADC-1 and ADC-5, differ by nine mutations in their protein sequence. ADC-5 hydrolyses cefoxitin better than ADC-1 and the opposite is true for ceftazidime. We produced single and combined mutations in ADC-5 and characterized the variants microbiologically and biochemically to determine which amino acid residues are involved in the hydrolysis of β-lactam antibiotics in this family of β-lactamases. Methods Site-directed mutagenesis, with blaADC-5 as a source of DNA, was used to generate nine single mutated and three combined mutated enzymes. The proteins (wild-type and derivatives) were then expressed in isogenic conditions in Escherichia coli. MICs of β-lactams were determined using Etest strips. ADC-1, ADC-5, ADC-5-P167S and ADC-5-P167S/D242G/Q163K/G342R were also purified and the kinetic parameters determined for ceftazidime, cefoxitin, cefalotin and ampicillin. Results Single mutations did not significantly convert the hydrolysis spectrum of the ADC-5 enzyme into that of the ADC-1 enzyme, although among all studied mutants only the quadruple mutant (ADC-5-P167S/D242G/Q163K/G342R) displayed microbiological and biochemical properties consistent with those of ADC-1. Conclusions Although some single mutations are known to affect cefepime hydrolysis in ADC-type β-lactamases, little is known about ceftazidime and cefoxitin hydrolysis in this family of β-lactamases. Hydrolysis of these antibiotics appears to be positively and negatively affected, respectively, by the Q163K, P167S, D242G and G342R amino acid replacements.

A method using dual process columns of Matrix Solid Phase Dispersion (MSPD) and Solid Phase Extraction (SPE) has been developed for extracting and cleaning-up of nine triazine herbicides (ametryn, atrazine, cyanazine, prometryn, propazine, simazine, simetryn, terbuthylazine and terbutryn) in seaweed samples. Under optimized conditions, samples were blended with 2g of octasilyl-derivatized silica (C8) and transferred into an SPE cartridge containing ENVI-Carb II/PSA (0.5/0.5 g) as a clean up co-sorbent. Then the dispersed sample was washed with 10 mL of n-hexane and triazines were eluted with 20 mL ethyl acetate and 5 mL acetonitrile. Finally the extract was concentrated to dryness, re-constituted with 1 mL methanol:water (1:1) and injected into the HPLC-DAD system. The linearity of the calibration curves was excellent in matrix matched standards, and yielded the coefficients of determination0.995 for all the target analytes. The recoveries ranged from 75% to 100% with relative standard deviations lower than 7%. The achieved LOQs (10 µg kg(-1)) for all triazines under study permits to ensure proper determination at the maximum allowed residue levels set in the European Union Legislation. Samples of three seaweeds were subjected to the procedure proving the suitability of MSPD method for the analysis of triazines in different seaweeds samples.

The use of anticoagulants has increased in recent times as a method for controlling rodent populations. However, this increased use also provokes accidental and intentional ingestion for both animals and humans, triggering poisoning of non-target organisms. In the present report, a clinical case of secondary-poisoning of birds with anticoagulant rodenticides, which took place after a general rodenticide treatment in an Ornithological Zoological Park, is described. Three birds died as a result and samples were submitted to the Veterinary Hospital in Lugo (Galicia, NW Spain). After necropsy, samples of the birds, together with molluscs and faeces, were submitted to the Toxicology Unit of Caceres (Extremadura, W Spain) in order to detect possible chemicals. Results from HPLC analyses revealed the presence of the rodenticides difenacoum and brodifacoum. The present report shows that the risk of secondary exposure resulting from the scavenging of molluscs is likely to be significant. The potential routes of uptake by invertebrates include the consumption of rodent faeces, rodent carcases, the ingestion of soil-bound residues, and the direct consumption of poison baits. Irene de la Casa-Resino was supported by fellowship PRE09001 from Department of Employment, Enterprise and Innovation (Gobierno de Extremadura, Spain) SI

A model describing the low temperature solid state phase transformation kinetics observed in a metal organic framework by differential scanning calorimetry (DSC) at several cooling rates is modified so that the reaction rate is now expressed as a function of time and temperature. Thus, when applied to ramp data, the new model exactly matches the former one but, additionally, it allows to explain isothermal data. The new model is tested for primary crystallizations of two polymers from the molten state, using DSC data, cooling ramp experiments at several cooling rates and isothermal experiments. Good fittings were obtained at all the varied experimental conditions with both polymers. The model makes use of three fitting parameters with physical meaning: an upper critical temperature, T c , an energy barrier, and a reaction-order, n + 1. Additionally, and previously to perform the kinetic fitting, the dependence of the time to the maximum crystallization rate peak on the isothermal temperature was investigated. That dependence was found to follow a simple model which makes use of two parameters related to the limits of the temperature range in which the crystallization may occur. The polymers used in this work were a commercial extruded polyamide and pristine syndiotactic polypropylene.

It has been shown that introducing carbon nanotubes (CNT) into a polymer matrix has a beneficial effect on thermal stability of composites. While the specific effects noted differ depending on many parameters, the general trend is an increase in the degradation temperature and reduction in the degradation rate, quantified as the mass decline over time. The purpose of this study is to evaluate how CNTs influence the main degradation process of composites made with polystyrene containing 2, 3 and 5 % of CNTs. Thermogravimetric experiments are performed, with nitrogen purge, at multiple linear heating ramps. The effects of the nanotubes on the degradation of polystyrene are evaluated. Insightful kinetic parameters were obtained for the main process making use of a recently developed model, which is adapted to the thermogravimetric context. The model allows the means to separate the main process from other processes, which could interfere with the kinetic analysis, and also subtract the residual mass, which could produce an apparent stabilizing effect. The main degradation process is clearly stabilized by the presence of nanotubes, although the stabilization is more pronounced at the lowest of the filler contents considered. Clear effects of nanotubes on kinetic parameters were observed.

The aim of this study was to determine heavy metal reference levels for risk assessment studies. For this purpose, the levels of lead, cadmium, copper and zinc were determined in liver tissues of wild boars sampled in NW Spain. The mean values were 0.383, 0.326, 23.50 and 56.86 mg/kg dried weight, respectively. In general, the levels detected were similar to or lower than the levels reported in literature. This study not only provides a useful baseline for biomonitoring the levels of the analyzed contaminants in wildlife in NW Spain, it also helps to understand the effects of gender on the levels of these elements. Similar to studies performed in other geographical regions, no significant gender-related differences could be detected. Although differences were not significant, the levels of zinc, cadmium and lead were modestly higher in males (55.78, 0.346 and 0.424 mg/kg, respectively) compared to females (45.25, 0.305 and 0.341 mg/kg). Our results indicate that, although gender did not significantly affect heavy metal uptake and toxicokinetics of contaminants in wild boars, these effects could vary between species, populations, organs, and elements. It is therefore essential to investigate gender-related differences for each species. © 2017 Elsevier Inc.

A simple method based on solid-phase extraction combined with liquid chromatography for simultaneous determination of nine triazine herbicides (ametryn, atrazine, cyanazine, prometryn, propazine, simazine, simetryn, terbuthylazine, and terbutryn) in surface water samples was developed and validated. Under optimized conditions, 50 mL of water sample was pumped through the Oasis HLB cartridge, and triazines were eluted with 3 mL acetone. Finally the extract was concentrated to dryness, reconstituted with 1 mL methanol : water (1 : 1) and injected into the HPLC-DAD system. The stability of the herbicides on the cartridges at −18 and 4°C was also evaluated, and the recoveries obtained after three weeks of storage were satisfactory for all compounds. The analytical features of the proposed method were satisfactory: repeatability and intermediate precision were −1) were adequately allowing the determination of these compounds at the levels requested by the 2008/105/EC Directive. Finally, this method was applied to the analysis of 50 seawater samples from Galicia (northwest Spain).

Triazines and their degradation products are transported to the aquatic environment, and once there, the probability to reach the marine environment is very high. In this paper, solid phase extraction (SPE) and extraction by matrix solid phase dispersion (MSPD) to analyse nine triazines (ametryn, atrazine, cyanazine, prometryn, propazine, simazine, simetryn, terbuthylazine and terbutryn) and eight degradation products (desethylatrazine, desethyldesisopropylatrazine, desethyl-2-hydroxyatrazine, desethylterbuthylazine, desisopropylatrazine, desisopropyl-2-hydroxyatrazine, 2-hydroxyatrazine and 2-hidroxyterbuthylazine) in seawater and marine sediments samples were used. The analysis was carried out using liquid chromatography with tandem mass spectrometry (LC-ESI-MS/MS). The methods were optimized and validated to achieve a selective and sensitive determination of the analytes from different sample, regardless of its complexity. Under the optimum conditions, the proposed methods provided adequate limits of quantification (0.05–0.45 μg L−1 and 0.23–4.26 μg kg−1 in seawater and marine sediments, respectively). Intra- and inter-day relative standard deviation were below 1.41% for all compounds. Recoveries were evaluated, and acceptable values that ranged from 87.5–99.4 and 60.9–99.7% for the seawater and sediment samples, respectively, were obtained. The proposed methods were applied to the analysis of the target compounds in seawater samples and marine sediments from a coastal area of Galicia (NW of Spain).

International audience; Neutron-rich light nuclei and their reactions play an important role for the creation of chemical elements. Here, data from a Coulomb dissociation experiment on $^{20,21}$N are reported. Relativistic $^{20,21}$N ions impinged on a lead target and the Coulomb dissociation cross section was determined in a kinematically complete experiment. Using the detailed balance theorem, the $^{19}\mathrm{N}(\mathrm{n},\gamma)^{20}\mathrm{N}$ and $^{20}\mathrm{N}(\mathrm{n},\gamma)^{21}\mathrm{N}$ excitation functions and thermonuclear reaction rates have been determined. The $^{19}\mathrm{N}(\mathrm{n},\gamma)^{20}\mathrm{N}$ rate is up to a factor of 5 higher at $T

Purpose . This work shows an effective methodology to characterize the creep–recovery behavior of silicones before their application in podiatry. The aim is to characterize, model and compare the creep–recovery properties of different types of silicone used in podiatry orthotics. Methods . Creep–recovery phenomena of silicones used in podiatry orthotics is characterized by dynamic mechanical analysis (DMA). Silicones provided by Herbitas are compared by observing their viscoelastic properties by Functional Data Analysis (FDA) and nonlinear regression. The relationship between strain and time is modeled by fixed and mixed effects nonlinear regression to compare easily and intuitively podiatry silicones. Results . Functional ANOVA and Kohlrausch–Willians–Watts (KWW) model with fixed and mixed effects allows us to compare different silicones observing the values of fitting parameters and their physical meaning. The differences between silicones are related to the variations of breadth of creep–recovery time distribution and instantaneous deformation-permanent strain. Nevertheless, the mean creep-relaxation time is the same for all the studied silicones. Silicones used in palliative orthoses have higher instantaneous deformation-permanent strain and narrower creep–recovery distribution. Conclusions . The proposed methodology based on DMA, FDA and nonlinear regression is an useful tool to characterize and choose the proper silicone for each podiatry application according to their viscoelastic properties.

Objectives Biofilm formation and bacterial adherence are important requirements for persistence, multidrug resistance and infection. The d-amino acids play a role as modulators of bacterial growth and persistence, though their ability to inhibit biofilms is much debated. In this study, we analysed the effects of 18 different d-amino acids on the pathogens Acinetobacter baumannii and Pseudomonas aeruginosa. Methods In vitro assays were carried out to analyse the effect of d-amino acids on bacterial growth, biofilm formation/disassembly, capacity to attach to eukaryotic cells and cellular death. In addition, in vivo assays were performed in mice, using experimental models of sepsis and pneumonia. Results Biofilm formation was inhibited in A. baumannii by d-His, d-Cys and d-Trp (35%-86%) at 2 mM and in P. aeruginosa by d-Cys, d-Trp and d-Tyr (10%-30%) at 4 mM. Attachment to the A549 human alveolar cells was reduced in A. baumannii by d-Cys, d-His, d-Met, d-Val and d-Ser, and in P. aeruginosa by d-Arg and d-Trp. Growth was inhibited in A. baumannii by d-Cys and d-Trp, and in P. aeruginosa by d-Trp. In virulence assays, incubation of alveolar cells infected with P. aeruginosa with d-Cys, d-Trp and d-Arg reduced cell death (56%-45%). However, no significant effect of d-amino acids was observed in vivo. Conclusions Some d-amino acids can inhibit bacterial growth, biofilm formation and adherence to eukaryotic cells in A. baumannii and P. aeruginosa, and showed a protective effect against infection of alveolar cells with P. aeruginosa. Despite the fact that some considerable protection was observed in mice, survival differences between treated and control groups were not statistically significant.

Synergy between colistin and the signal peptidase inhibitor MD3 was tested against isogenic mutants and clinical pairs of Acinetobacter baumannii isolates. Checkerboard assays and growth curves showed synergy against both colistin-susceptible strains (fractional inhibitory concentration index [FIC index ] = 0.13 to 0.24) and colistin-resistant strains with mutations in pmrB and phosphoethanolamine modification of lipid A (FIC index = 0.14 to 0.25) but not against colistin-resistant Δ lpx strains with loss of lipopolysaccharide (FIC index = 0.75 to 1). A colistin/MD3 combination would need to be targeted to strains with specific colistin resistance mechanisms.