Structural Basis of Enzymatic Activity for the Ferulic Acid Decarboxylase (FADase) from Enterobacter sp. Px6-4

Microbial ferulic acid decarboxylase (FADase) catalyzes the transformation of ferulic acid to 4-hydroxy-3-methoxystyrene (4-vinylguaiacol) via non-oxidative decarboxylation. Here we report the crystal structures of the Enterobacter sp. Px6-4 FADase and the enzyme in complex with substrate analogues. Our analyses revealed that FADase possessed a half-opened bottom β-barrel with the catalytic pocket located between the middle of the core β-barrel and the helical bottom. Its structure shared a high degree of similarity with members of the phenolic acid decarboxylase (PAD) superfamily. Structural analysis revealed that FADase catalyzed reactions by an “open-closed” mechanism involving a pocket of 8×8×15 Å dimension on the surface of the enzyme. The active pocket could directly contact the solvent and allow the substrate to enter when induced by substrate analogues. Site-directed mutagenesis showed that the E134A mutation decreased the enzyme activity by more than 60%, and Y21A and Y27A mutations abolished the enzyme activity completely. The combined structural and mutagenesis results suggest that during decarboxylation of ferulic acid by FADase, Trp25 and Tyr27 are required for the entering and proper orientation of the substrate while Glu134 and Asn23 participate in proton transfer

Left Ventricular Noncompaction:Anatomical Phenotype or Distinct Cardiomyopathy?

The present study was funded by the Souter Charitable Foundation and the Chest, Heart and Stroke Scotland Charity. Dr. Weir-McCall is supported by the Wellcome Trust through the Scottish Translational Medicine and Therapeutics Initiative (Grant no. WT 085664) in the form of a Clinical Research Fellowship.Background:  There is considerable overlap between left ventricular noncompaction (LVNC) and other cardiomyopathies. LVNC has been reported in up to 40% of the general population, raising questions about whether it is a distinct pathological entity, a remodeling epiphenomenon, or merely an anatomical phenotype. Objectives:   The authors determined the prevalence and predictors of LVNC in a healthy population using 4 cardiac magnetic resonance imaging diagnostic criteria. Methods:   Volunteers >40 years of age (N = 1,651) with no history of cardiovascular disease (CVD), a 10-year risk of CVD < 20%, and a B-type natriuretic peptide level greater than their gender-specific median underwent magnetic resonance imaging scan as part of the TASCFORCE (Tayside Screening for Cardiac Events) study. LVNC ratios were measured on the horizontal and vertical long axis cine sequences. All individuals with a noncompaction ratio of ≥2 underwent short axis systolic and diastolic LVNC ratio measurements, and quantification of noncompacted and compacted myocardial mass ratios. Those who met all 4 criteria were considered to have LVNC. Results:  Of 1,480 participants analyzed, 219 (14.8%) met ≥1 diagnostic criterion for LVNC, 117 (7.9%) met 2 criteria, 63 (4.3%) met 3 criteria, and 19 (1.3%) met all 4 diagnostic criteria. There was no difference in demographic or allometric measures between those with and without LVNC. Long axis noncompaction ratios were the least specific, with current diagnostic criteria positive in 219 (14.8%), whereas the noncompacted to compacted myocardial mass ratio was the most specific, only being met in 61 (4.4%). Conclusions:   A significant proportion of an asymptomatic population free from CVD satisfy all currently used cardiac magnetic resonance imaging diagnostic criteria for LVNC, suggesting that those criteria have poor specificity for LVNC, or that LVNC is an anatomical phenotype rather than a distinct cardiomyopathy.Publisher PDFPeer reviewe

Late Cretaceous continental and marine vertebrate assemblages of the Laño Quarry (Basque-Cantabrian Region, Iberian Peninsula): an update

The vertebrate-bearing beds of the Laño quarry (Condado de Treviño) are among the most relevant sites from the Late Cretaceous of Europe. Geologically, Laño and the adjacent region are set on the southern limb of the South-Cantabrian Synclinorium (SE Basque-Cantabrian Region, northern Iberian Peninsula). The Laño sites were discovered in 1984; thousands of bones and teeth, including microfossils, have been collected during the prospection in the field and excavation campaigns. The vertebrate remains occur at two different stratigraphic horizons within a continental to shallow marine succession of Late Campanian-Maastrichtian age. The lower horizon contains the Laño 1 and Laño 2 sites, whereas the upper horizon contains the Albaina site. In the Laño sites, three fossiliferous beds (called L1A, L1B and L2) are known within an alluvial system composed mainly of fluvial sands and silts. The sedimentary structures are consistent with channel areas within an extensive braided river system. Based mainly on stratigraphic correlations, the fluvial beds of Laño are regarded as Late Campanian to Early Maastrichtian in age. These deposits have yielded a very diverse vertebrate assemblage, which consists of nearly 40 species, including actinopterygians, lissamphibians, lepidosaurs, turtles, crocodyliforms, dinosaurs, pterosaurs, and mammals. Seven genera and ten species have been erected to date in Laño. With reference to the marine vertebrate association of Albaina, it consists of at least 37 species, including sharks and rays, actinopterygians, mosasaurids, and plesiosaurs. Two genera and species of rhinobatoids (family indet.) and two new species of rhinobatids have been erected in Albaina. The fossil association indicates a Late (but not latest) Maastrichtian age. Recently, isolated turtle and dinosaur fossils have been discovered in the sublittoral beds of Albaina. The Laño quarry is one of the most noteworthy Campanian-Maastrichtian vertebrate localities of Europe by its taxonomic diversity, and provides useful information about the composition and affinities of both continental and marine vertebrate faunas from the latest Cretaceous of southwestern Europe.Los niveles con fósiles de vertebrados de la cantera de Laño (Condado de Treviño) se cuentan entre los más importantes del Cretácico Superior de Europa. Desde un punto de vista geológico, Laño y la región adyacente forman parte del flanco sur del Sinclinorio Subcantábrico (SE de la Región Vasco-Cantábrica). El descubrimiento de los niveles fosilíferos remonta a 1984; las prospecciones de campo y las campañas de excavación han proporcionado miles de huesos y dientes, incluyendo microfósiles. Los restos de vertebrados aparecen en dos horizontes estratigráficos diferentes formando parte de una sucesión continental a marina litoral de edad Campaniense superior a Maastrichtiense. El horizonte inferior contiene los yacimientos de Laño 1 y Laño 2, mientras que el superior contiene solo uno: Albaina. En los yacimientos de Laño, se reconocen tres niveles fosilíferos (llamados L1A, L1B y L2) formados en el seno de un sistema aluvial compuesto por arenas y limos fluviales. Las estructuras sedimentarias indican áreas de canal dentro de un sistema trenzado muy extendido. Según las correlaciones estratigráficas, los depósitos fluviales de Laño son de edad Campaniense superior a Maastrichtiense inferior. Estos depósitos han proporcionado una asociación muy diversa de vertebrados, que consiste en cerca de 40 especies, incluyendo actinopterigios, lisanfibios, lepidosaurios, tortugas, cocodrilos, dinosaurios, pterosaurios y mamíferos. En Laño se han definido hasta el momento siete géneros y diez especies. Por lo que respecta a la asociación de vertebrados marinos de Albaina, se han reconocido al menos 37 especies, que incluyen tiburones y rayas, actinopterigios, mosasaurios y plesiosaurios. Hasta la fecha se han definido en Albaina cuatro nuevos rinobatoideos: dos especies de rinobátidos y dos géneros y especies cuya familia es indeterminada. La asociación fósil es de edad Maastrichtiense superior no terminal. Recientemente se ha descrito el hallazgo de restos fósiles aislados de tortugas y dinosaurios en los niveles marinos de Albaina. La cantera de Laño es una de las localidades de vertebrados más destacadas del Campaniense-Maastrichtiense por su diversidad taxonómica, y proporciona información relevante sobre la composición y afinidades de las faunas de vertebrados continentales y marinos del Cretácico final del suroeste de Europa

A Phylogenetic Perspective on the Evolution of Mediterranean Teleost Fishes

The Mediterranean Sea is a highly diverse, highly studied, and highly impacted biogeographic region, yet no phylogenetic reconstruction of fish diversity in this area has been published to date. Here, we infer the timing and geographic origins of Mediterranean teleost species diversity using nucleotide sequences collected from GenBank. We assembled a DNA supermatrix composed of four mitochondrial genes (12S ribosomal DNA, 16S ribosomal DNA, cytochrome c oxidase subunit I and cytochrome b) and two nuclear genes (rhodopsin and recombination activating gene I), including 62% of Mediterranean teleost species plus 9 outgroups. Maximum likelihood and Bayesian phylogenetic and dating analyses were calibrated using 20 fossil constraints. An additional 124 species were grafted onto the chronogram according to their taxonomic affinity, checking for the effects of taxonomic coverage in subsequent diversification analyses. We then interpreted the time-line of teleost diversification in light of Mediterranean historical biogeography, distinguishing non-endemic natives, endemics and exotic species. Results show that the major Mediterranean orders are of Cretaceous origin, specifically ∼100–80 Mya, and most Perciformes families originated 80–50 Mya. Two important clade origin events were detected. The first at 100–80 Mya, affected native and exotic species, and reflects a global diversification period at a time when the Mediterranean Sea did not yet exist. The second occurred during the last 50 Mya, and is noticeable among endemic and native species, but not among exotic species. This period corresponds to isolation of the Mediterranean from Indo-Pacific waters before the Messinian salinity crisis. The Mediterranean fish fauna illustrates well the assembly of regional faunas through origination and immigration, where dispersal and isolation have shaped the emergence of a biodiversity hotspot

Phase 1, placebo-controlled, dose escalation trial of chicory root extract in patients with osteoarthritis of the hip or knee

<p>Abstract</p> <p>Background</p> <p>Extracts of chicory root have anti-inflammatory properties <it>in vitro </it>and in animal models of arthritis. The primary objective of this investigator-initiated, Phase 1, placebo-controlled, double blind, dose-escalating trial was to determine the safety and tolerability of a proprietary bioactive extract of chicory root in patients with osteoarthritis (OA). Secondary objectives were to assess effects on the signs and symptoms of this disorder.</p> <p>Methods</p> <p>Individuals greater than 50 years of age with OA of the hip or knee were eligible for trial entry. A total of 40 patients were enrolled in 3 cohorts and were treated with escalating chicory doses of 600 mg/day, 1200 mg/day and 1800 mg/day for 1 month. The ratio of active treatment to placebo was 5:3 in cohorts 1 and 2 (8 patients) each and 16:8 in cohort 3 (24 patients). Safety evaluations included measurement of vital signs and routine lab tests at baseline and the end of the treatment period. Efficacy evaluations at baseline and final visits included self-assessment questionnaires and measurement of the 25-foot walking time.</p> <p>Results</p> <p>In the highest dose cohort, 18 patients who completed treatment per protocol were analyzed for efficacy. In this group, 13 patients showed at least 20% improvement in the defined response domains of pain, stiffness and global assessment: 9 of 10 (90%) patients randomized to active treatment with chicory and 4 of 8 (50%) patients randomized to placebo (P = 0.06). In general, the treatment was well-tolerated. Only one patient who was treated with the highest dose of chicory had to discontinue treatment due to an adverse event.</p> <p>Conclusions</p> <p>The results of this pilot study suggest that a proprietary bioactive extract of chicory root has a potential role in the management of OA and merits further investigation. Clinicaltrials.gov identifier: NCT 01010919.</p

Sensitivity of the IceCube Detector to Astrophysical Sources of High Energy Muon Neutrinos

We present the results of a Monte-Carlo study of the sensitivity of the planned IceCube detector to predicted fluxes of muon neutrinos at TeV to PeV energies. A complete simulation of the detector and data analysis is used to study the detector's capability to search for muon neutrinos from sources such as active galaxies and gamma-ray bursts. We study the effective area and the angular resolution of the detector as a function of muon energy and angle of incidence. We present detailed calculations of the sensitivity of the detector to both diffuse and pointlike neutrino emissions, including an assessment of the sensitivity to neutrinos detected in coincidence with gamma-ray burst observations. After three years of datataking, IceCube will have been able to detect a point source flux of E^2*dN/dE = 7*10^-9 cm^-2s^-1GeV at a 5-sigma significance, or, in the absence of a signal, place a 90% c.l. limit at a level E^2*dN/dE = 2*10^-9 cm^-2s^-1GeV. A diffuse E-2 flux would be detectable at a minimum strength of E^2*dN/dE = 1*10^-8 cm^-2s^-1sr^-1GeV. A gamma-ray burst model following the formulation of Waxman and Bahcall would result in a 5-sigma effect after the observation of 200 bursts in coincidence with satellite observations of the gamma-rays.Comment: 33 pages, 13 figures, 6 table

Late cretaceous continental and marine vertebrate assemblages of the Laño Quarry (Basque-Cantabrian Region, Iberian Peninsula): An update

The vertebrate-bearing beds of the Laño quarry (Condado de Treviño) are among the most relevant sites from the Late Cretaceous of Europe. Geologically, Laño and the adjacent region are set on the southern limb of the South-Cantabrian Synclinorium (SE Basque-Cantabrian Region, northern Iberian Peninsula). The Laño sites were discovered in 1984; thousands of bones and teeth, including microfossils, have been collected during the prospection in the field and excavation campaigns. The vertebrate remains occur at two different stratigraphic horizons within a continental to shallow marine succession of Late Campanian-Maastrichtian age. The lower horizon contains the Laño 1 and Laño 2 sites, whereas the upper horizon contains the Albaina site. In the Laño sites, three fossiliferous beds (called L1A, L1B and L2) are known within an alluvial system composed mainly of fluvial sands and silts. The sedimentary structures are consistent with channel areas within an extensive braided river system. Based mainly on stratigraphic correlations, the fluvial beds of Laño are regarded as Late Campanian to Early Maastrichtian in age. These deposits have yielded a very diverse vertebrate assemblage, which consists of nearly 40 species, including actinopterygians, lissamphibians, lepidosaurs, turtles, crocodyliforms, dinosaurs, pterosaurs, and mammals. Seven genera and ten species have been erected to date in Laño. With reference to the marine vertebrate association of Albaina, it consists of at least 37 species, including sharks and rays, actinopterygians, mosasaurids, and plesiosaurs. Two genera and species of rhinobatoids (family indet.) and two new species of rhinobatids have been erected in Albaina. The fossil association indicates a Late (but not latest) Maastrichtian age. Recently, isolated turtle and dinosaur fossils have been discovered in the sublittoral beds of Albaina. The Laño quarry is one of the most noteworthy Campanian-Maastrichtian vertebrate localities of Europe by its taxonomic diversity, and provides useful information about the composition and affinities of both continental and marine vertebrate faunas from the latest Cretaceous of southwestern EuropeLos niveles con fósiles de vertebrados de la cantera de Laño (Condado de Treviño) se cuentan entre los más importantes del Cretácico Superior de Europa. Desde un punto de vista geológico, Laño y la región adyacente forman parte del flanco sur del Sinclinorio Subcantábrico (SE de la Región Vasco-Cantábrica). El descubrimiento de los niveles fosilíferos remonta a 1984; las prospecciones de campo y las campañas de excavación han proporcionado miles de huesos y dientes, incluyendo microfósiles. Los restos de vertebrados aparecen en dos horizontes estratigráficos diferentes formando parte de una sucesión continental a marina litoral de edad Campaniense superior a Maastrichtiense. El horizonte inferior contiene los yacimientos de Laño 1 y Laño 2, mientras que el superior contiene solo uno: Albaina. En los yacimientos de Laño, se reconocen tres niveles fosilíferos (llamados L1A, L1B y L2) formados en el seno de un sistema aluvial compuesto por arenas y limos fluviales. Las estructuras sedimentarias indican áreas de canal dentro de un sistema trenzado muy extendido. Según las correlaciones estratigráficas, los depósitos fluviales de Laño son de edad Campaniense superior a Maastrichtiense inferior. Estos depósitos han proporcionado una asociación muy diversa de vertebrados, que consiste en cerca de 40 especies, incluyendo actinopterigios, lisanfibios, lepidosaurios, tortugas, cocodrilos, dinosaurios, pterosaurios y mamíferos. En Laño se han definido hasta el momento siete géneros y diez especies. Por lo que respecta a la asociación de vertebrados marinos de Albaina, se han reconocido al menos 37 especies, que incluyen tiburones y rayas, actinopterigios, mosasaurios y plesiosaurios. Hasta la fecha se han definido en Albaina cuatro nuevos rinobatoideos: dos especies de rinobátidos y dos géneros y especies cuya familia es indeterminada. La asociación fósil es de edad Maastrichtiense superior no terminal. Recientemente se ha descrito el hallazgo de restos fósiles aislados de tortugas y dinosaurios en los niveles marinos de Albaina. La cantera de Laño es una de las localidades de vertebrados más destacadas del Campaniense-Maastrichtiense por su diversidad taxonómica, y proporciona información relevante sobre la composición y afinidades de las faunas de vertebrados continentales y marinos del Cretácico final del suroeste de EuropaField research in the Laño quarry has been supported by financial aids of the Ministerio de Economía y Competitividad (MINECO projects CGL2010-18851/BTE and CGL2013-47521-P), Ministerio de Ciencia e Innovación (CGL2007-64061/BTE), Ministerio de Ciencia y Tecnología (CGL2004-02338/BTE, BTE2001-0185-C02-01, BOS2000-1369), Ministerio de Educación y Ciencia (Acción Integrada hispano-francesa 201-B), Diputación Foral de Álava/Arabako Foru Aldundia (93/23, 95/A23), Gobierno Vasco/Eusko Jaurlaritza (IT834-13, IT-320-10, IT-361-07, GV121.310-4/87), Universidad del País Vasco/Euskal Herriko Unibertsitatea (9/ UPV00121.310-15303/2003), Centre National de la Recherche Scientifique (CNRS, France), Institut National des Sciences de l’Univers (INSU), The Dinosaur Society (research grant 1997) and the National Geographic Research (grant #6597-99). This work is part of a palaeontological collaboration (Convenio específico de colaboración) between the Universidad del País Vasco/EHU, the Centre National de la Recherche Scientifique (CNRS, France) and the Muséum National d’Histoire Naturelle (MNHN, Paris). Contribution ISEM n° 2015-04

Loss of Nrf2 abrogates the protective effect of Keap1 down regulation in a preclinical model of cutaneous squamous cell carcinoma

Cutaneous squamous cell carcinomas (cSCC) are the most common and highly mutated human malignancies, challenging identification of driver mutations and targeted therapies. Transcription factor NF-E2 p45-related factor 2 (Nrf2) orchestrates a cytoprotective inducible program, which counteracts the damaging effects of solar UV radiation, the main etiological factor in cSCC development. Downregulation of Kelch-like ECH-associated protein 1 (Keap1), a Cullin-3/Rbx1 ubiquitin ligase substrate adaptor protein, which mediates the ubiquitination and proteasomal degradation of Nrf2, has a strong protective effect in a preclinical model of cSCC. However, in addition to Nrf2, Keap1 affects ubiquitination of other proteins in the carcinogenesis process, including proteins involved in inflammation and DNA damage repair. Here, we generated Keap1(flox/flox) SKH-1 hairless mice in which Nrf2 is disrupted (Keap1(flox/flox)/Nrf2(−/−)) and subjected them chronically to solar-simulated UV radiation. We found that the incidence, multiplicity and burden of cSCC that form in Keap1(flox/flox)/Nrf2(−/−) mice are much greater than in their Keap1(flox/flox)/Nrf2(+/+) counterparts, establishing Nrf2 activation as the protection mediator. Our findings further imply that inhibition of Nrf2 globally, a strategy proposed for cancer treatment, is unlikely to be beneficial