Distribution of Complex and Core Lipids within New Hyperthermophilic Members of the Archaea Domain

Core and complex lipids of several new hyperthermophilic archaeal isolates were analyzed. The organisms belong to the Sulfolobales,Archaeoglobus, Pyrobaculum, and Methanococcus. A detailed structural investigation of complex lipids of Pyrobaculum species is reported. The different lipid structures are of help for a rapid and simple phylogenetic classification of the new isolates. They are in agreement with the classification based on other features

Current trends on seaweeds: Looking at chemical composition, phytopharmacology, and cosmetic applications

Seaweeds have received huge interest in recent years given their promising potentialities. Their antioxidant, anti-inflammatory, antitumor, hypolipemic, and anticoagulant effects are among the most renowned and studied bioactivities so far, and these effects have been increasingly associated with their content and richness in both primary and secondary metabolites. Although primary metabolites have a pivotal importance such as their content in polysaccharides (fucoidans, agars, carragenans, ulvans, alginates, and laminarin), recent data have shown that the content in some secondary metabolites largely determines the effective bioactive potential of seaweeds. Among these secondary metabolites, phenolic compounds feature prominently. The present review provides the most remarkable insights into seaweed research, specifically addressing its chemical composition, phytopharmacology, and cosmetic applications.We would like to thank the University of Aveiro and FCT/MCT for their financial support for the QOPNA Research Unit (FCT UID/QUI/00062/2019) and the cE3c Center (UID/BIA/00329/2013) through national founds, and where applicable, co-financed by the FEDER within the PT2020 Partnership Agreement. Martins N. would like to thank the Portuguese Foundation for Science and Technology (FCT-Portugal) for the strategic project ref. UID/BIM/04293/2013 and "NORTE2020-Programa Operacional Regional do Norte" (NORTE-01-0145-FEDER-000012). Acknowledgments: We would like to thank the University of Aveiro and FCT/MCT for their financial support for the QOPNA Research Unit (FCT UID/QUI/00062/2019) and the cE3c Center (UID/BIA/00329/2013) through national founds, and where applicable, co-financed by the FEDER within the PT2020 Partnership Agreement. Martins N. would like to thank the Portuguese Foundation for Science and Technology (FCT–Portugal) for the strategic project ref. UID/BIM/04293/2013 and “NORTE2020—Programa Operacional Regional do Norte” (NORTE-01-0145-FEDER-000012)

Passenger Dimensions in Sustainable Multimodal Mobility Services

Seamless integration of air segment in the overall multimodal mobility chain is a key challenge to provide more efficient and sustainable transport services. Technology advances offer a unique opportunity to build a new generation of transport services able to match the evolving expectations and needs of society as a whole. In this context, the passenger-centric approach represents a method to inform the design of future mobility services, supporting quality of life, security and services to citizens traveling across Europe. Relying on the concepts of inclusive design, context of use and task analysis, in this article, we present a comprehensive methodological framework for the analysis of passenger characteristics to elicit features and requirements for future multimodal mobility services, including air leg, that are relevant from the perspective of passengers. The proposed methodology was applied to a series of specific use cases envisaged for three time horizons, 2025, 2035 and 2050, in the context of a European research project. Then, passenger-focused key performance indicators and related metrics were derived to be included in a validation step, with the aim of assessing the extent of benefit for passengers that can be achieved in the forecasted scenarios. The results of the study demonstrate the relevance of human variability in the design of public services, as well as the feasibility of personalized performance assessment of mobility services

SNi from SN2: a front-face mechanism ‘synthase’ engineered from a retaining hydrolase

SNi or SNi-like mechanisms, in which leaving group departure and nucleophile approach occur on the same ‘front’ face, have been observed previously experimentally and computationally in both the chemical and enzymatic (glycosyltransferase) substitution reactions of α-glycosyl electrophiles. Given the availability of often energetically comparable competing pathways for substitution (SNi vs SN1 vs SN2) the precise modulation of this archetypal reaction type should be feasible. Here, we show that the drastic engineering of a protein that catalyzes substitution, a retaining β-glycosidase (from Sulfolobus solfataricus SSβG), apparently changes the mode of reaction from “SN2” to “SNi”. Destruction of the nucleophilic Glu387 of SSβG-WT through Glu387Tyr mutation (E387Y) created a catalyst (SSβG-E387Y) with lowered but clear transglycosylation substitution activity with activated substrates, altered substrate and reaction preferences and hence useful synthetic (‘synthase’) utility by virtue of its low hydrolytic activity with unactivated substrates. Strikingly, the catalyst still displayed retaining β stereoselectivity, despite lacking a suitable nucleophile; pH-activity profile, mechanism-based inactivators and mutational analyses suggest that SSβG-E387Y operates without either the use of nucleophile or general acid/base residues, consistent with a SNi or SNi-like mechanism. An x-ray structure of SSβG-E387Y and subsequent metadynamics simulation suggest recruitment of substrates aided by a π-sugar interaction with the introduced Tyr387 and reveal a QM/MM free energy landscape for the substitution reaction catalyzed by this unnatural enzyme similar to those of known natural, SNi-like glycosyltransferase (GT) enzymes. Proton flight from the putative hydroxyl nucleophile to the developing p-nitrophenoxide leaving group of the substituted molecule in the reactant complex creates a hydrogen bond that appears to crucially facilitate the mechanism, mimicking the natural mechanism of SNi-GTs. An oxocarbenium ion-pair minimum along the reaction pathway suggests a step-wise SNi-like DN*ANss rather than a concerted SNi DNAN mechanism. This first observation of a front face mechanism in a β-retaining glycosyl transfer enzyme highlights, not only that unusual SNi reaction pathways may be accessed through direct engineering of catalysts with suitable environments, but also suggests that ‘β-SNi’ reactions are also feasible for glycosyl transfer enzymes and the more widespread existence of SNi or SNi-like mechanism in nature

Biochemical and structural characterisation of a haloalkane dehalogenase from a marine Rhodobacteraceae

types: Journal Article; Research Support, Non-U.S. Gov'tCopyright © 2014 Federation of European Biochemical Societies. Published by Elsevier B.V. NOTICE: This is the author’s version of a work accepted for publication by Elsevier. Changes resulting from the publishing process, including peer review, editing, corrections, structural formatting and other quality control mechanisms, may not be reflected in this document. Changes may have been made to this work since it was submitted for publication. A definitive version was subsequently published in FEBS Letters Vol. 588, Issue 9, pp. 1616 – 1622 DOI: 10.1016/j.febslet.2014.02.056A putative haloalkane dehalogenase has been identified in a marine Rhodobacteraceae and subsequently cloned and over-expressed in Escherichia coli. The enzyme has highest activity towards the substrates 1,6-dichlorohexane, 1-bromooctane, 1,3-dibromopropane and 1-bromohexane. The crystal structures of the enzyme in the native and product bound forms reveal a large hydrophobic active site cavity. A deeper substrate binding pocket defines the enzyme preference towards substrates with longer carbon chains. Arg136 at the bottom of the substrate pocket is positioned to bind the distal halogen group of extended di-halogenated substrates.Wellcome TrustEPSRCHRMUniversity of ExeterBBSR

Human milk and mucosal lacto- and galacto-N-biose synthesis by transgalactosylation and their prebiotic potential in Lactobacillus species

Lacto-N-biose (LNB) and galacto-N-biose (GNB) are major building blocks of free oligosaccharides and glycan moieties of glyco-complexes present in human milk and gastrointestinal mucosa. We have previously characterized the phospho-β-galactosidase GnbG from Lactobacillus casei BL23 that is involved in the metabolism of LNB and GNB. GnbG has been used here in transglycosylation reactions, and it showed the production of LNB and GNB with N-acetylglucosamine and N-acetylgalactosamine as acceptors, respectively. The reaction kinetics demonstrated that GnbG can convert 69 ± 4 and 71 ± 1 % of o-nitrophenyl-β-D-galactopyranoside into LNB and GNB, respectively. Those reactions were performed in a semi-preparative scale, and the synthesized disaccharides were purified. The maximum yield obtained for LNB was 10.7 ± 0.2 g/l and for GNB was 10.8 ± 0.3 g/l. NMR spectroscopy confirmed the molecular structures of both carbohydrates and the absence of reaction byproducts, which also supports that GnbG is specific for β1,3-glycosidic linkages. The purified sugars were subsequently tested for their potential prebiotic properties using Lactobacillus species. The results showed that LNB and GNB were fermented by the tested strains of L. casei, Lactobacillus rhamnosus (except L. rhamnosus strain ATCC 53103), Lactobacillus zeae, Lactobacillus gasseri, and Lactobacillus johnsonii. DNA hybridization experiments suggested that the metabolism of those disaccharides in 9 out of 10 L. casei strains, all L. rhamnosus strains and all L. zeae strains tested relies upon a phospho-β-galactosidase homologous to GnbG. The results presented here support the putative role of human milk oligosaccharides for selective enrichment of beneficial intestinal microbiota in breast-fed infants

Effects of airbag deployment - Lesions, epidemiology, and management

Injuries caused by airbag deployment are described. The epidemiology of various lesions caused by air bag is reporte

Air bags and the skin.

Air bags, fitted in the majority of new automobiles, are safety devices activated when a sudden deceleration causes the ignition of a propellant cartridge containing sodium azide. The bag is inflated by nitrogen liberated during the combustion. Deployment releases various high-temperature gases, including nitrogen and carbon dioxide, and produces sodium hydroxide, a highly irritant alkaline substance. In about 7%-8% of cases, air bags cause dermatologic injuries such as traumatic lesions, irritant dermatitis, and chemical and thermal burns. Nondermatologic lesions, such as ocular damage (alkali keratitis, corneal abrasions), ear lesions, bone fractures, and contusive damage can also be caused by air bag deployment