Structural and biochemical characterization of the exopolysaccharide deacetylase Agd3 required for Aspergillus fumigatus biofilm formation

The exopolysaccharide galactosaminogalactan (GAG) is an important virulence factor of the fungal pathogen Aspergillus fumigatus. Deletion of a gene encoding a putative deacetylase, Agd3, leads to defects in GAG deacetylation, biofilm formation, and virulence. Here, we show that Agd3 deacetylates GAG in a metal-dependent manner, and is the founding member of carbohydrate esterase family CE18. The active site is formed by four catalytic motifs that are essential for activity. The structure of Agd3 includes an elongated substrate-binding cleft formed by a carbohydrate binding module (CBM) that is the founding member of CBM family 87. Agd3 homologues are encoded in previously unidentified putative bacterial exopolysaccharide biosynthetic operons and in other fungal genomes. The exopolysaccharide galactosaminogalactan (GAG) is an important virulence factor of the fungal pathogen Aspergillus fumigatus. Here, the authors study an A. fumigatus enzyme that deacetylates GAG in a metal-dependent manner and constitutes a founding member of a new carbohydrate esterase family.Bio-organic Synthesi

The Electronics and Data Acquisition System of the DarkSide Dark Matter Search

It is generally inferred from astronomical measurements that Dark Matter (DM) comprises approximately 27\% of the energy-density of the universe. If DM is a subatomic particle, a possible candidate is a Weakly Interacting Massive Particle (WIMP), and the DarkSide-50 (DS) experiment is a direct search for evidence of WIMP-nuclear collisions. DS is located underground at the Laboratori Nazionali del Gran Sasso (LNGS) in Italy, and consists of three active, embedded components; an outer water veto (CTF), a liquid scintillator veto (LSV), and a liquid argon (LAr) time projection chamber (TPC). This paper describes the data acquisition and electronic systems of the DS detectors, designed to detect the residual ionization from such collisions

Structural and functional insight into the mechanism of an alkaline exonuclease from Laribacter hongkongensis

Alkaline exonuclease and single-strand DNA (ssDNA) annealing proteins (SSAPs) are key components of DNA recombination and repair systems within many prokaryotes, bacteriophages and virus-like genetic elements. The recently sequenced β-proteobacterium Laribacter hongkongensis (strain HLHK9) encodes putative homologs of alkaline exonuclease (LHK-Exo) and SSAP (LHK-Bet) proteins on its 3.17 Mb genome. Here, we report the biophysical, biochemical and structural characterization of recombinant LHK-Exo protein. LHK-Exo digests linear double-stranded DNA molecules from their 5′-termini in a highly processive manner. Exonuclease activities are optimum at pH 8.2 and essentially require Mg2+ or Mn2+ ions. 5′-phosphorylated DNA substrates are preferred over dephosphorylated ones. The crystal structure of LHK-Exo was resolved to 1.9 Å, revealing a ‘doughnut-shaped’ toroidal trimeric arrangement with a central tapered channel, analogous to that of λ-exonuclease (Exo) from bacteriophage-λ. Active sites containing two bound Mg2+ ions on each of the three monomers were located in clefts exposed to this central channel. Crystal structures of LHK-Exo in complex with dAMP and ssDNA were determined to elucidate the structural basis for substrate recognition and binding. Through structure-guided mutational analysis, we discuss the roles played by various active site residues. A conserved two metal ion catalytic mechanism is proposed for this class of alkaline exonucleases

The Familial Intracranial Aneurysm (FIA) study protocol

BACKGROUND: Subarachnoid hemorrhage (SAH) due to ruptured intracranial aneurysms (IAs) occurs in about 20,000 people per year in the U.S. annually and nearly half of the affected persons are dead within the first 30 days. Survivors of ruptured IAs are often left with substantial disability. Thus, primary prevention of aneurysm formation and rupture is of paramount importance. Prior studies indicate that genetic factors are important in the formation and rupture of IAs. The long-term goal of the Familial Intracranial Aneurysm (FIA) Study is to identify genes that underlie the development and rupture of intracranial aneurysms (IA). METHODS/DESIGN: The FIA Study includes 26 clinical centers which have extensive experience in the clinical management and imaging of intracerebral aneurysms. 475 families with affected sib pairs or with multiple affected relatives will be enrolled through retrospective and prospective screening of potential subjects with an IA. After giving informed consent, the proband or their spokesperson invites other family members to participate. Each participant is interviewed using a standardized questionnaire which covers medical history, social history and demographic information. In addition blood is drawn from each participant for DNA isolation and immortalization of lymphocytes. High- risk family members without a previously diagnosed IA undergo magnetic resonance angiography (MRA) to identify asymptomatic unruptured aneurysms. A 10 cM genome screen will be performed to identify FIA susceptibility loci. Due to the significant mortality of affected individuals, novel approaches are employed to reconstruct the genotype of critical deceased individuals. These include the intensive recruitment of the spouse and children of deceased, affected individuals. DISCUSSION: A successful, adequately-powered genetic linkage study of IA is challenging given the very high, early mortality of ruptured IA. Design features in the FIA Study that address this challenge include recruitment at a large number of highly active clinical centers, comprehensive screening and recruitment techniques, non-invasive vascular imaging of high-risk subjects, genome reconstruction of dead affected individuals using marker data from closely related family members, and inclusion of environmental covariates in the statistical analysis

Higher harmonic anisotropic flow measurements of charged particles in Pb-Pb collisions at 2.76 TeV

We report on the first measurement of the triangular $v_3$, quadrangular $v_4$, and pentagonal $v_5$ charged particle flow in Pb-Pb collisions at 2.76 TeV measured with the ALICE detector at the CERN Large Hadron Collider. We show that the triangular flow can be described in terms of the initial spatial anisotropy and its fluctuations, which provides strong constraints on its origin. In the most central events, where the elliptic flow $v_2$ and $v_3$ have similar magnitude, a double peaked structure in the two-particle azimuthal correlations is observed, which is often interpreted as a Mach cone response to fast partons. We show that this structure can be naturally explained from the measured anisotropic flow Fourier coefficients.Comment: 10 pages, 4 figures, published version, figures at http://aliceinfo.cern.ch/ArtSubmission/node/387

Reconstructed May-July mean maximum temperature since 1745 AD based on tree-ring width of Pinus tabulaeformis in Qianshan Mountain, China

A tree-ring-width chronology of Pinus tabulaeformis was developed from the Qianshan Mountain, Liaoning province, northeastern China. Based on the correlation between the ring width and instrumental data, a transfer function was designed and the May-July mean maximum temperature (MMT) from 1745 to 2012 was reconstructed. The reconstruction explains 42.7% of the instrumental variance during the calibration period (41.7% after adjusting for the loss of the degrees of freedom). The reconstructed MMT is similar to several observed MMT series and the temperature index in north-central China, which indicated that the decrease in summer temperatures in the 20th century was a large scale phenomenon. The reconstruction also showed that high MMT values corresponded to historical drought events in Liaoning. In addition, a spatial Correlation analyses revealed that the MMT reconstruction is regionally representative. Significant 128.2-, 64.1-, 18.6-, 3.46-, 3.19-, 2.43-, 2.15- and 2.10-year cycles were detected in the reconstructed MMT series from Qianshan Mountain.</p

Poly aluminum chloride (PAC) enhanced formation of aerobic granules:Coupling process between physicochemical–biochemical effects

Transforming conventional flocculent sludge to aerobic granular sludge is drawing increasing global interest in a quest for an efficient and innovative technology in wastewater treatment. However, long start-up time and low granule stability are the main challenging issues for its application. In this study, long-term and short-term PAC feeding strategies were applied in parallel to enhance the sludge granulation and with the aim to figure out the temporal effect of PAC during the whole process. Nevertheless, both of them identified to allow a rapid start-up formation of aerobic granules with better performances in physicochemical characteristics. More extracellular polymeric substances (EPS) were secreted in all PAC-fed reactors, especially for polysaccharides. By using enzymatic hydrolysis, factors which account for the flocculating ability and stability of granules were identified. Notably, the correlation between them was also revealed. Based on these findings, dosage of PAC played a positive role mainly during the start-up period (first 15&nbsp;days) of the aerobic granules formation, extending its dosing time made no significant sense. The granules formed under this condition were the result of physicochemical&ndash;biochemical effects coupling process.</p

Comparison between luminescence and radiocarbon dating of lateQuaternary loess from the Ili Basin in Central Asia

Dust depositions are critical archives for understanding interior aridification and westerly climatic changes in Central Asia. Accurate and reliable dating of loess is very important for interpreting and correlating environmental records. There remains a disparity between luminescence ages and radiocarbon dating of late Quaternary loess from the Ili Basin in Central Asia. In this study, we establish a closely spaced quartz optically stimulated luminescence (OSL) chronology for the 20.5-m-thick Nilka loess section in the Ili Basin. Based on OSL ages, two intervals of higher mass accumulation rate occurred at 49&ndash;43&nbsp;ka and 24&ndash;14&nbsp;ka. We further compare these OSL ages with 23 accelerator mass spectrometry (AMS) 14C ages of bulk organic matter. The results indicate that the OSL and radiocarbon ages agree well for ages younger than ca. 25 14C cal&nbsp;ka&nbsp;BP. However, beyond 30&nbsp;cal&nbsp;ka&nbsp;BP, there is no consistent increase in AMS 14C age with depth, while the OSL ages continue to increase. These differences confirm the observation that the AMS 14C ages obtained using conventional acid&ndash;base&ndash;acid (ABA) pretreatment are severely underestimated in other terrestrial deposits in Central Asia, which could be due to 2&ndash;4% modern carbon contamination. However, OSL dating is applicable for constructing an accurate chronology beyond 30&nbsp;cal&nbsp;ka&nbsp;BP. We suggest caution when interpreting paleoenvironmental changes based on radiocarbon ages older than 25&nbsp;cal&nbsp;ka&nbsp;BP.</p