The essential role of water molecules in the reaction mechanism of protein O-Fucosyltransferase 2

Protein O-fucosyltransferase 2 (PoFUT2) is an inverting glycosyltransferase (GT) that fucosylates thrombospondin repeats (TSRs) from group 1 and 2. PoFUT2 recognizes a large and diverse number of TSRs through a dynamic network of water-mediated interactions. By X-ray structural studies of C. elegans PoFUT2 complexed to a TSR of group 2, we demonstrate that this GT recognizes similarly the 3D structure of TSRs from both groups 1 and 2. Its active site is highly exposed to the solvent, suggesting that water molecules might also play an essential role in the fucosylation mechanism. We applied QM/MM methods using human PoFUT2 as a model, and found that HsPoFUT2 follows a classical S(N)2 reaction mechanism in which water molecules contribute to a great extent in facilitating the release of the leaving pyrophosphate unit, causing the H transfer from the acceptor nucleophile (Thr/Ser) to the catalytic base, which is the last event in the reaction. This demonstrates the importance of water molecules not only in recognition of the ligands but also in catalysis

Biomass-modulated fire dynamics during the last glacial-interglacial transition at the central pyrenees (Spain)

Understanding long-term fire ecology is essential for current day interpretation of ecosystem fire responses. However palaeoecology of fire is still poorly understood, especially at high-altitude mountain environments, despite the fact that these are fire-sensitive ecosystems and their resilience might be affected by changing fire regimes. We reconstruct wildfire occurrence since the Lateglacial (14.7. cal. ka BP) to the Mid-Holocene (6. cal. ka BP) and investigate the climate-fuel-fire relationships in a sedimentary sequence located at the treeline in the Central Spanish Pyrenees. Pollen, macro- and micro-charcoal were analysed for the identification of fire events (FE) in order to detect vegetation post-fire response and to define biomass-fire interactions. mean fire intervals (mfi) reduced since the Lateglacial, peaking at 9-7.7. cal. ka BP while from 7.7 to 6. cal. ka BP no fire is recorded. We hypothesise that Early Holocene maximum summer insolation, as climate forcing, and mesophyte forest expansion, as a fuel-creating factor, were responsible for accelerating fire occurrence in the Central Pyrenees treeline. We also found that fire had long-lasting negative effects on most of the treeline plant communities and that forest contraction from 7.7. cal. ka BP is likely linked to the ecosystem's threshold response to high fire frequencies.This research has been funded by the projects DINAMO (CGL2009-07992) (funding EGPF — grant ref. BES-2010-038593 and MSC), DINAMO2 (CGL2012-33063), ARAFIRE (2012 GA LC 064), GRACCIE-CONSOLIDER (CSD2007-00067). GGR was funded by the Juan de la Cierva Program (grant ref. JCI2009-04345) and JAE-Doc CSIC Program, LLM was supported by a postdoctoral MINT fellowship funded by the Institute for the Environment (Brunel University), AMC is a Ramón y Cajal fellow (ref: RYC-2008-02431), APS holds a grant funded by the Aragon Government (ref. 17030G/5423/480072/14003) and JAE holds a grant funded by the Basque Country Government (BFI-2010-5)

Medical versus surgical approach to initial treatment in septic arthritis: A single spanish center’s 8-year experience

Objective The aim of this study was to compare the functional results of 2 different procedure types, medical or surgical used in treating native joint septic arthritis. Methods In this cohort study, we reviewed the clinical registries of patients admitted to a single third-level hospital with the diagnosis of septic arthritis during the period of January 1, 2008, to January 31, 2016. Results A total of 63 cases of septic arthritis were identified in which the initial approach for 49 patients was medical (arthrocentesis), whereas the initial approach for 14 patients was surgical (arthroscopy or arthrotomy). Of the 49 patients who received initial medical treatment (IMT), 15 patients (30%) later required surgical treatment because of poor progress. The median age of the patients was 60 (SD, 18) years. The group who received IMT were older than those who received initial surgical treatment (median, 64 years [interquartile range {IQR}, 54–76 years], vs. 48 years [IQR, 30–60 years]). There was a larger percentage of male patients in the surgical group (78% vs. 42% [p = 0.018]). Thirty percent of the medical group had been receiving corticosteroid treatment (p = 0.018). Results of complete recovery of joint functionality showed no significant differences after 1 year (68% with MT vs. 67% with ST, p = 0.91). Both groups had similar symptom duration until diagnosis, duration of antibiotic therapy (median, 30 days [IQR, 28–49 days], vs. 29.5 days [IQR, 27–49] days), and mortality rate (3 in the medical group). Conclusions The results of the study show that initial surgical treatment in patients with native joint septic arthritis is not superior to IMT. However, half of the patients with shoulder and hip infections treated with IMT eventually required surgical intervention, suggesting that perhaps this should be the preferred initial approach in these cases

Cambios en el estado nutricional, composición corporal y sintomatología asociada en pacientes hospitalizados sometidos a trasplante de médula ósea: estudio longitudinal prospectivo

Introducción: Los pacientes sometidos a Trasplante de Células Madre Hematopoyéticas o trasplante de médula padecen varias complicaciones nutricionales. El objetivo del estudio fue realizar una descripción prospectiva en estos pacientes. Material y métodos: Se reclutaron 14 pacientes con una edad media de 48,0±9,8 años. Resultados: El 28,6% padecía sobrepeso, el 14,3% obesidad y el 57,1% tenía un peso normal, con una evolución entre -0,3±0,3 kg/m2 (normopeso) hasta -3,1±0,2 kg/m2 (obesidad). Se observó una pérdida de peso variable, de hasta 7,3±0,7% en pacientes con mayor Índice de Masa Corporal (IMC). La composición corporal también empeoró al alta, con una evolución de la circunferencia braquial de -1,7±0,4 cm en trasplantes alogénicos y -2±4,5 cm en trasplantes autólogos. Un 42,9% de hombres y 28,6% de mujeres eran dados de alta con un Índice de Masa Libre de Grasa (IMLG) por debajo de las recomendaciones, incrementándose desde el ingreso en hombres (desde un 14,3%). Se observa una pérdida de fuerza muscular, en trasplantes alogénicos de -5,0±1,5 kg en el caso de hombres, y -3,0±0,5 kg en mujeres; en autólogos, de -5 kg y -4 kg respectivamente. Sobre síntomas, al inicio existía una alta prevalencia de vómitos (71,4%), náuseas (42,9%) o saciedad temprana (57,1%); durante la hospitalización, destaca la saciedad temprana (92,9%), náuseas (71,4%), vómitos (71,4%), disgeusia (57,1%), diarrea (50%) y anorexia (50%). Conclusiones: Los pacientes admitidos para trasplante de médula ósea están aparentemente bien nutridos, y existe un deterioro durante la hospitalización; parece adecuado implementar estrategias dietéticas durante la hospitalización para optimizar la ingesta y prevenir la malnutrición

Endocannabinoid levels in peripheral blood mononuclear cells of multiple sclerosis patients treated with dimethyl fumarate.

The endocannabinoid system (ECS), a signalling network with immunomodulatory properties, is a potential therapeutic target in multiple sclerosis (MS). Dimethyl fumarate (DMF) is an approved drug for MS whose mechanism of action has not been fully elucidated; the possibility exists that its therapeutic effects could imply the ECS. With the aim of studying if DMF can modulate the ECS, the endocannabinoids 2-arachidonoylglycerol (2-AG), anandamide (AEA), oleoylethanolamide (OEA) and palmitoylethanolamide (PEA) were determined by liquid chromatography-mass spectrometry in peripheral blood mononuclear cells from 21 healthy donors (HD) and 32 MS patients at baseline and after 12 and 24 months of DMF treatment. MS patients presented lower levels of 2-AG and PEA compared to HD. 2-AG increased at 24 months, reaching HD levels. AEA and PEA remained stable at 12 and 24 months. OEA increased at 12 months and returned to initial levels at 24 months. Patients who achieved no evidence of disease activity (NEDA3) presented the same modulation over time as EDA3 patients. PEA was modulated differentially between females and males. Our results show that the ECS is dysregulated in MS patients. The increase in 2-AG and OEA during DMF treatment suggests a possible role of DMF in ECS modulation.post-print1392 K

Strategies for GHG mitigation in Mediterranean cropping systems. A review

In this review we aimed to synthetize and analyze the most promising GHGs mitigation strategies for Mediterranean cropping systems. A description of most relevant measures, based on the best crop choice and management by farmers (i.e., agronomical practices), was firstly carried out. Many of these measures can be also efficient in other climatic regions, but here we provide particular results and discussion of their efficiencies for Mediterranean cropping systems. An integrated assessment of management practices on mitigating each component of the global warming potential (N2O and CH4 emissions and C sequestration) of production systems considering potential side-effects of their implementation allowed us to propose the best strategies to abate GHG emissions, while sustaining crop yields and mitigating other sources of environmental pollution (e.g. nitrate leaching and ammonia volatilization)

Strategies for greenhouse gas emissions mitigation in Mediterranean agriculture: A review

[EN] An integrated assessment of the potential of different management practices for mitigating specific components of the total GHG budget (N2O and CH4 emissions and C sequestration) of Mediterranean agrosystems was performed in this study. Their suitability regarding both yield and environmental (e.g. nitrate leaching and ammonia volatilization) sustainability, and regional barriers and opportunities for their implementation were also considered. Based on its results best strategies to abate GHG emissions in Mediterranean agro-systems were proposed. Adjusting N fertilization to crop needs in both irrigated and rain-fed systems could reduce N2O emissions up to 50% compared with a non-adjusted practice. Substitution of N synthetic fertilizers by solid manure can be also implemented in those systems, and may abate N2O emissions by about 20% under Mediterranean conditions, with additional indirect benefits associated to energy savings and positive effects in crop yields. The use of urease and nitrification inhibitors enhances N use efficiency of the cropping systems and may mitigate N2O emissions up to 80% and 50%, respectively. The type of irrigation may also have a great mitigation potential in the Mediterranean region. Drip-irrigated systems have on average 80% lower N2O emissions than sprinkler systems and drip-irrigation combined with optimized fertilization showed a reduction in direct N2O emissions up to 50%. Methane fluxes have a relatively small contribution to the total GHG budget of Mediterranean crops, which can mostly be controlled by careful management of the water table and organic inputs in paddies. Reduced soil tillage, improved management of crop residues and agro-industry by-products, and cover cropping in orchards, are the most suitable interventions to enhance organic C stocks in Mediterranean agricultural soils. The adoption of the proposed agricultural practices will require farmers training. The global analysis of life cycle emissions associated to irrigation type (drip, sprinkle and furrow) and N fertilization rate (100 and 300 kg N ha(-1) yr(-1)) revealed that these factors may outweigh the reduction in GHG emissions beyond the plot scale. The analysis of the impact of some structural changes on top-down mitigation of GHG emissions revealed that 3-15% of N2O emissions could be suppressed by avoiding food waste at the end-consumer level. A 40% reduction in meat and dairy consumption could reduce GHG emissions by 20-30%. Reintroducing the Mediterranean diet (i.e. similar to 35% intake of animal protein) would therefore result in a significant decrease of GHG emissions from agricultural production systems under Mediterranean conditions. (C) 2016 Elsevier B.V. All rights reserved.The authors would like to thank the Spanish National R+D+i Plan (AGL2012-37815-C05-01, AGL2012-37815-C05-04) and very specifically the workshop held in December 2016 in Butron (Bizkaia) to synthesize the most promising measures to reduce N2O emissions from Spanish agricultural soils. BC3 is sponsored by the Basque Government. M. L. Cayuela thanks Fundacion Seneca for financing the project 19281/PI/14.Sanz-Cobeña, A.; Lassaletta, L.; Aguilera, E.; Del Prado, A.; Garnier, J.; Billen, G.; Iglesias, A.... (2017). Strategies for greenhouse gas emissions mitigation in Mediterranean agriculture: A review. Agriculture Ecosystems & Environment. 238:5-24. https://doi.org/10.1016/j.agee.2016.09.038S52423

ATTACK, a novel bispecific T cell-recruiting antibody with trivalent EGFR binding and monovalent CD3 binding for cancer immunotherapy

The redirection of T cell activity using bispecific antibodies is one of the most promising cancer immunotherapy approaches currently in development, but it is limited by cytokine storm-related toxicities, as well as the pharmacokinetics and tumor-penetrating capabilities of current bispecific antibody formats. Here, we have engineered the ATTACK (Asymmetric Tandem Trimerbody for T cell Activation and Cancer Killing), a novel T cell-recruiting bispecific antibody which combines three EGFR-binding single-domain antibodies (VHH; clone EgA1) with a single CD3-binding single-chain variable fragment (scFv; clone OKT3) in an intermediate molecular weight package. The two specificities are oriented in opposite directions in order to simultaneously engage cancer cells and T cell effectors, and thereby promote immunological synapse formation. EgA1 ATTACK was expressed as a homogenous, non-aggregating, soluble protein by mammalian cells and demonstrated an enhanced binding to EGFR, but not CD3, when compared to the previously characterized tandem bispecific antibody which has one EgA1 VHH and one OKT3 scFv per molecule. EgA1 ATTACK induced synapse formation and early signaling pathways downstream of TCR engagement at lower concentrations than the tandem VHH-scFv bispecific antibody. Furthermore, it demonstrated extremely potent, dose-dependent cytotoxicity when retargeting human T cells towards EGFR-expressing cells, with an efficacy over 15-fold higher than that of the tandem VHH-scFv bispecific antibody. These results suggest that the ATTACK is an ideal format for the development of the next-generation of T cell-redirecting bispecific antibodies