Sample records for abortus ure2 region

  1. Analysis of a Soluble (UreD:UreF:UreG)2 Accessory Protein Complex and its Interactions with Klebsiella aerogenes Urease by Mass Spectrometry

    PubMed Central

    Farrugia, Mark A.; Han, Linjie; Zhong, Yueyang; Boer, Jodi L.; Ruotolo, Brandon T.; Hausinger, Robert P.

    2013-01-01

    Maturation of the nickel-containing urease of Klebsiella aerogenes is facilitated by the UreD, UreF, and UreG accessory proteins along with the UreE metallo-chaperone. A fusion of the maltose binding protein and UreD (MBP-UreD) was co-isolated with UreF and UreG in a soluble complex possessing a (MBP-UreD:UreF:UreG)2 quaternary structure. Within this complex a UreF:UreF interaction was identified by chemical cross-linking of the amino termini of its two UreF protomers, as shown by mass spectrometry of tryptic peptides. A pre-activation complex was formed by the interaction of (MBP-UreD:UreF:UreG)2 and urease. Mass spectrometry of intact protein species revealed a pathway for synthesis of the urease pre-activation complex in which individual hetero-trimer units of the (MBP-UreD:UreF:UreG)2 complex bind to urease. Together, these data provide important new insights into the structures of protein complexes associated with urease activation. PMID:23797863

  2. Analysis of a Soluble (UreD:UreF:UreG)2 Accessory Protein Complex and Its Interactions with Klebsiella aerogenes Urease by Mass Spectrometry

    NASA Astrophysics Data System (ADS)

    Farrugia, Mark A.; Han, Linjie; Zhong, Yueyang; Boer, Jodi L.; Ruotolo, Brandon T.; Hausinger, Robert P.

    2013-09-01

    Maturation of the nickel-containing urease of Klebsiella aerogenes is facilitated by the UreD, UreF, and UreG accessory proteins along with the UreE metallo-chaperone. A fusion of the maltose binding protein and UreD (MBP-UreD) was co-isolated with UreF and UreG in a soluble complex possessing a (MBP-UreD:UreF:UreG)2 quaternary structure. Within this complex a UreF:UreF interaction was identified by chemical cross-linking of the amino termini of its two UreF protomers, as shown by mass spectrometry of tryptic peptides. A pre-activation complex was formed by the interaction of (MBP-UreD:UreF:UreG)2 and urease. Mass spectrometry of intact protein species revealed a pathway for synthesis of the urease pre-activation complex in which individual hetero-trimer units of the (MBP-UreD:UreF:UreG)2 complex bind to urease. Together, these data provide important new insights into the structures of protein complexes associated with urease activation.

  3. UreE-UreG Complex Facilitates Nickel Transfer and Preactivates GTPase of UreG in Helicobacter pylori*♦

    PubMed Central

    Yang, Xinming; Li, Hongyan; Lai, Tsz-Pui; Sun, Hongzhe

    2015-01-01

    The pathogenicity of Helicobacter pylori relies heavily on urease, which converts urea to ammonia to neutralize the stomach acid. Incorporation of Ni2+ into the active site of urease requires a battery of chaperones. Both metallochaperones UreE and UreG play important roles in the urease activation. In this study, we demonstrate that, in the presence of GTP and Mg2+, UreG binds Ni2+ with an affinity (Kd) of ∼0.36 μm. The GTPase activity of Ni2+-UreG is stimulated by both K+ (or NH4+) and HCO3− to a biologically relevant level, suggesting that K+/NH4+ and HCO3− might serve as GTPase elements of UreG. We show that complexation of UreE and UreG results in two protein complexes, i.e. 2E-2G and 2E-G, with the former being formed only in the presence of both GTP and Mg2+. Mutagenesis studies reveal that Arg-101 on UreE and Cys-66 on UreG are critical for stabilization of 2E-2G complex. Combined biophysical and bioassay studies show that the formation of 2E-2G complex not only facilitates nickel transfer from UreE to UreG, but also enhances the binding of GTP. This suggests that UreE might also serve as a structural scaffold for recruitment of GTP to UreG. Importantly, we demonstrate for the first time that UreE serves as a bridge to grasp Ni2+ from HypA, subsequently donating it to UreG. The study expands our horizons on the molecular details of nickel translocation among metallochaperones UreE, UreG, and HypA, which further extends our knowledge on the urease maturation process. PMID:25752610

  4. Molecular landscape of the interaction between the urease accessory proteins UreE and UreG.

    PubMed

    Merloni, Anna; Dobrovolska, Olena; Zambelli, Barbara; Agostini, Federico; Bazzani, Micaela; Musiani, Francesco; Ciurli, Stefano

    2014-09-01

    Urease, the most efficient enzyme so far discovered, depends on the presence of nickel ions in the catalytic site for its activity. The transformation of inactive apo-urease into active holo-urease requires the insertion of two Ni(II) ions in the substrate binding site, a process that involves the interaction of four accessory proteins named UreD, UreF, UreG and UreE. This study, carried out using calorimetric and NMR-based structural analysis, is focused on the interaction between UreE and UreG from Sporosarcina pasteurii, a highly ureolytic bacterium. Isothermal calorimetric protein-protein titrations revealed the occurrence of a binding event between SpUreE and SpUreG, entailing two independent steps with positive cooperativity (Kd1=42±9μM; Kd2=1.7±0.3μM). This was interpreted as indicating the formation of the (UreE)2(UreG)2 hetero-oligomer upon binding of two UreG monomers onto the pre-formed UreE dimer. The molecular details of this interaction were elucidated using high-resolution NMR spectroscopy. The occurrence of SpUreE chemical shift perturbations upon addition of SpUreG was investigated and analyzed to establish the protein-protein interaction site. The latter appears to involve the Ni(II) binding site as well as mobile portions on the C-terminal and the N-terminal domains. Docking calculations based on the information obtained from NMR provided a structural basis for the protein-protein contact site. The high sequence and structural similarity within these protein classes suggests a generality of the interaction mode among homologous proteins. The implications of these results on the molecular details of the urease activation process are considered and analyzed. Copyright © 2014 Elsevier B.V. All rights reserved.

  5. Structure of UreG/UreF/UreH Complex Reveals How Urease Accessory Proteins Facilitate Maturation of Helicobacter pylori Urease

    PubMed Central

    Fong, Yu Hang; Wong, Ho Chun; Yuen, Man Hon; Lau, Pak Ho; Chen, Yu Wai; Wong, Kam-Bo

    2013-01-01

    Urease is a metalloenzyme essential for the survival of Helicobacter pylori in acidic gastric environment. Maturation of urease involves carbamylation of Lys219 and insertion of two nickel ions at its active site. This process requires GTP hydrolysis and the formation of a preactivation complex consisting of apo-urease and urease accessory proteins UreF, UreH, and UreG. UreF and UreH form a complex to recruit UreG, which is a SIMIBI class GTPase, to the preactivation complex. We report here the crystal structure of the UreG/UreF/UreH complex, which illustrates how UreF and UreH facilitate dimerization of UreG, and assembles its metal binding site by juxtaposing two invariant Cys66-Pro67-His68 metal binding motif at the interface to form the (UreG/UreF/UreH)2 complex. Interaction studies revealed that addition of nickel and GTP to the UreG/UreF/UreH complex releases a UreG dimer that binds a nickel ion at the dimeric interface. Substitution of Cys66 and His68 with alanine abolishes the formation of the nickel-charged UreG dimer. This nickel-charged UreG dimer can activate urease in vitro in the presence of the UreF/UreH complex. Static light scattering and atomic absorption spectroscopy measurements demonstrated that the nickel-charged UreG dimer, upon GTP hydrolysis, reverts to its monomeric form and releases nickel to urease. Based on our results, we propose a mechanism on how urease accessory proteins facilitate maturation of urease. PMID:24115911

  6. The Function of UreB in Klebsiella aerogenes Urease†

    PubMed Central

    Carter, Eric L.; Boer, Jodi L.; Farrugia, Mark A.; Flugga, Nicholas; Towns, Christopher L.; Hausinger, Robert P.

    2011-01-01

    Urease from Klebsiella aerogenes is composed of three subunits (UreA, UreB, and UreC) which assemble into a (UreABC)3 quaternary structure. UreC harbors the dinuclear nickel active site, whereas the functions of UreA and UreB remain unknown. UreD and UreF accessory proteins previously were suggested to reposition UreB and increase exposure of the nascent urease active site, thus facilitating metallocenter assembly. In this study, cells were engineered to separately produce (UreAC)3 or UreB, and the purified proteins were characterized. Monomeric UreB spontaneously binds to the trimeric heterodimer of UreA plus UreC to form (UreABC*)3 apoprotein, as shown by gel filtration chromatography, integration of electrophoretic gel band intensities, and mass spectrometry. Similar to authentic urease apoprotein, active enzyme is produced by incubation of (UreABC*)3 with Ni2+ and bicarbonate. Conversely, UreBΔ1-19, lacking the 19 residue potential hinge and tether to UreC, does not form a complex with (UreAC)3 and yields negligible levels of active enzyme when incubated under activation conditions with (UreAC)3. Comparison of activities and nickel contents for (UreAC)3, (UreABC*)3, and (UreABC)3 samples treated with Ni2+ and bicarbonate and then desalted indicates that UreB facilitates efficient incorporation of the metal into the active site and protects the bound metal from chelation. Amylose resin pull-down studies reveal that MBP-UreD (a fusion of maltose binding protein with UreD) forms complexes with (UreABC)3, (UreAC)3, and UreB in vivo, but not in vitro. By contrast, MBP-UreD does not form an in vivo complex with UreBΔ1-19. The soluble MBP-UreD:UreF:UreG complex binds in vitro to (UreABC)3, but not to (UreAC)3 or UreB. Together these data demonstrate that UreB facilitates the interaction of urease with accessory proteins during metallocenter assembly, with the N-terminal hinge and tether region being specifically required for this process. In addition to its role in

  7. Characterization of the functional role of nucleotides within the URE2 IRES element and the requirements for eIF2A-mediated repression.

    PubMed

    Reineke, Lucas C; Merrick, William C

    2009-12-01

    Cap-independent initiation of translation is thought to promote protein synthesis on some mRNAs during times when cap-dependent initiation is down-regulated. However, the mechanism of cap-independent initiation is poorly understood. We have previously reported the secondary structure within the yeast minimal URE2 IRES element. In this study, we sought to investigate the mechanism of internal initiation in yeast by assessing the functional role of nucleotides within the minimal URE2 IRES element, and delineating the cis-sequences that modulate levels of internal initiation using a monocistronic reporter vector. Furthermore, we compared the eIF2A sensitivity of the URE2 IRES element with some of the invasive growth IRES elements using DeltaeIF2A yeast. We found that the stability of the stem-loop structure within the minimal URE2 IRES element is not a critical determinant of optimal IRES activity, and the downstream sequences that modulate URE2 IRES-mediated translation can be defined to discrete regions within the URE2 coding region. Repression of internal initiation on the URE2 minimal IRES element by eIF2A is not dependent on the stability of the secondary structure within the URE2 IRES element. Our data also indicate that eIF2A-mediated repression is not specific to the URE2 IRES element, as both the GIC1 and PAB1 IRES elements are repressed by eIF2A. These data provide valuable insights into the mRNA requirements for internal initiation in yeast, and insights into the mechanism of eIF2A-mediated suppression.

  8. Hermes Transposon Mutagenesis Shows [URE3] Prion Pathology Prevented by a Ubiquitin-Targeting Protein: Evidence for Carbon/Nitrogen Assimilation Cross-Talk and a Second Function for Ure2p in Saccharomyces cerevisiae.

    PubMed

    Edskes, Herman K; Mukhamedova, Maryam; Edskes, Bouke K; Wickner, Reed B

    2018-05-16

    [URE3] is an amyloid-based prion of Ure2p, a regulator of nitrogen catabolism. While most variants of the [URE3] prion are toxic, mild variants that only slightly slow growth are more widely studied. The existence of several anti-prion systems suggests that some components may be protecting cells from potential detrimental effects of mild [URE3] variants. Our extensive Hermes transposon mutagenesis showed that disruption of YLR352W dramatically slows growth of [URE3-1] strains. Ylr352wp is an F-box protein, directing selection of substrates for ubiquitination by a cullin-containing E- 3 ligase. For efficient ubiquitylation, cullin-dependent E 3 ubiquitin ligases must be NEDDylated, modified by a ubiquitin-related peptide called NEDD8 (Rub1p in yeast). Indeed, we find that disruption of NEDDylation-related genes, RUB1, ULA1, UBA3 and UBC12 is also counterselected in our screen. We find that like ylr352w Δ [URE3] strains, ylr352w Δ ure2 Δ strains do not grow on non-fermentable carbon sources. Overexpression of Hap4p, a transcription factor stimulating expression of mitochondrial proteins, or mutation of GLN1 , encoding glutamine synthetase, allow growth of ylr352w Δ [URE3] strains on glycerol media. Supplying proline as a nitrogen source shuts off the nitrogen catabolite repression (NCR) function of Ure2p, but does not slow growth of ylr352w Δ strains, suggesting a distinct function of Ure2p in carbon catabolism. Also, gln1 mutations impair NCR, but actually relieve the growth defect of ylr352w Δ [URE3] and ylr352w Δ ure2 Δ strains, again showing that loss of NCR is not producing the growth defect and suggesting Ure2p has another function. YLR352W largely protects cells from the deleterious effects of otherwise mild [URE3] variants, or of a ure2 mutation (the latter a rarer event), and we name it LUG1 (lets [URE3]/ ure2 grow). Copyright © 2018, Genetics.

  9. Klebsiella aerogenes UreF: Identification of the UreG Binding Site and Role in Enhancing the Fidelity of Urease Activation†

    PubMed Central

    Boer, Jodi L.; Hausinger, Robert P.

    2012-01-01

    The Ni-containing active site of Klebsiella aerogenes urease is assembled through the concerted action of the UreD, UreE, UreF, and UreG accessory proteins. UreE functions as a metallochaperone that delivers Ni to a complex of UreD—UreF—UreG bound to urease apoprotein, with UreG serving as a GTPase during enzyme activation. The present study focuses on the role of UreF, previously proposed to act as a GTPase activating protein (GAP) of UreG. Sixteen conserved UreF surface residues that may play roles in protein:protein interactions were independently changed to Ala. When produced in the context of the entire urease gene cluster, cell-free extracts of nine site-directed mutants had less than 10% of the wild-type urease activity. Enrichment of the variant forms of UreF, as the UreE-F fusion proteins, uniformly resulted in co-purification of UreD and urease apoprotein; whereas UreG bound to only a subset of the species. Notably, reduced interaction with UreG correlated with the low activity mutants. The affected residues in UreF map to a distinct surface on the crystal structure, defining the UreG binding site. In contrast to the hypothesis that UreF is a GAP, the UreD—UreF—UreG—urease apoprotein complex containing K165A UreF exhibited significantly greater levels of GTPase activity than that containing the wild-type protein. Additional studies demonstrated the UreG GTPase activity was largely uncoupled from urease activation for the complex containing this UreF variant. Further experiments with these complexes provided evidence that UreF gates the GTPase activity of UreG to enhance the fidelity of urease metallocenter assembly, especially in the presence of the non-cognate metal Zn. PMID:22369361

  10. Mutational and Computational Evidence That a Nickel-Transfer Tunnel in UreD Is Used for Activation of Klebsiella aerogenes Urease.

    PubMed

    Farrugia, Mark A; Wang, Beibei; Feig, Michael; Hausinger, Robert P

    2015-10-20

    Nickel-containing urease from Klebsiella aerogenes requires four accessory proteins for proper active site metalation. The metallochaperone UreE delivers nickel to UreG, a GTPase that forms a UreD/UreF/UreG complex, which binds to urease apoprotein via UreD. Prior in silico analysis of the homologous, structurally characterized UreH/UreF/UreG complex from Helicobacter pylori identified a water tunnel originating at a likely nickel-binding motif in UreG, passing through UreF, and exiting UreH, suggestive of a role for the channel in providing the metal to urease apoprotein for its activation; however, no experimental support was reported for the significance of this tunnel. Here, specific variants were designed to disrupt a comparable 34.6 Å predicted internal tunnel, alternative channels, and surface sites for UreD. Cells producing a set of tunnel-disrupting variants of UreD exhibited greatly reduced urease specific activities, whereas other mutants had no appreciable effect on activity. Affinity pull-down studies of cell-free extracts from tunnel-disrupting mutant cultures showed no loss of UreD interactions with urease or UreF/UreG. The nickel contents of urease samples enriched from activity-deficient cultures were decreased, while zinc and iron incorporation increased. Molecular dynamics simulations revealed size restrictions in the internal channels of the UreD variants. These findings support the role of a molecular tunnel in UreD as a direct facilitator of nickel transfer into urease, illustrating a new paradigm in active site metallocenter assembly.

  11. Mutational analysis of the major soybean UreF paralogue involved in urease activation.

    PubMed

    Polacco, Joe C; Hyten, David L; Medeiros-Silva, Mônica; Sleper, David A; Bilyeu, Kristin D

    2011-06-01

    The soybean genome duplicated ∼14 and 45 million years ago and has many paralogous genes, including those in urease activation (emplacement of Ni and CO(2) in the active site). Activation requires the UreD and UreF proteins, each encoded by two paralogues. UreG, a third essential activation protein, is encoded by the single-copy Eu3, and eu3 mutants lack activity of both urease isozymes. eu2 has the same urease-negative phenotype, consistent with Eu2 being a single-copy gene, possibly encoding a Ni carrier. Unexpectedly, two eu2 alleles co-segregated with missense mutations in the chromosome 2 UreF paralogue (Ch02UreF), suggesting lack of expression/function of Ch14UreF. However, Ch02UreF and Ch14UreF transcripts accumulate at the same level. Further, it had been shown that expression of the Ch14UreF ORF complemented a fungal ureF mutant. A third, nonsense (Q2*) allelic mutant, eu2-c, exhibited 5- to 10-fold more residual urease activity than missense eu2-a or eu2-b, though eu2-c should lack all Ch02UreF protein. It is hypothesized that low-level activation by Ch14UreF is 'spoiled' by the altered missense Ch02UreF proteins ('epistatic dominant-negative'). In agreement with active 'spoiling' by eu2-b-encoded Ch02UreF (G31D), eu2-b/eu2-c heterozygotes had less than half the urease activity of eu2-c/eu2-c siblings. Ch02UreF (G31D) could spoil activation by Chr14UreF because of higher affinity for the activation complex, or because Ch02UreF (G31D) is more abundant than Ch14UreF. Here, the latter is favoured, consistent with a reported in-frame AUG in the 5' leader of Chr14UreF transcript. Translational inhibition could represent a form of 'functional divergence' of duplicated genes.

  12. Crystal Structures of Apo and Metal-Bound Forms of the UreE Protein from Helicobacter pylori: Role of Multiple Metal Binding Sites

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Shi, Rong; Munger, Christine; Asinas, Abdalin

    2010-10-22

    The crystal structure of the urease maturation protein UreE from Helicobacter pylori has been determined in its apo form at 2.1 {angstrom} resolution, bound to Cu{sup 2+} at 2.7 {angstrom} resolution, and bound to Ni{sup 2+} at 3.1 {angstrom} resolution. Apo UreE forms dimers, while the metal-bound enzymes are arranged as tetramers that consist of a dimer of dimers associated around the metal ion through coordination by His102 residues from each subunit of the tetramer. Comparison of independent subunits from different crystal forms indicates changes in the relative arrangement of the N- and C-terminal domains in response to metal binding.more » The improved ability of engineered versions of UreE containing hexahistidine sequences at either the N-terminal or C-terminal end to provide Ni{sup 2+} for the final metal sink (urease) is eliminated in the H102A version. Therefore, the ability of the improved Ni{sup 2+}-binding versions to deliver more nickel is likely an effect of an increased local concentration of metal ions that can rapidly replenish transferred ions bound to His102.« less

  13. Mutational analysis of the major soybean UreF paralogue involved in urease activation

    PubMed Central

    Polacco, Joe C.; Hyten, David L.; Medeiros-Silva, Mônica; Sleper, David A.; Bilyeu, Kristin D.

    2011-01-01

    The soybean genome duplicated ∼14 and 45 million years ago and has many paralogous genes, including those in urease activation (emplacement of Ni and CO2 in the active site). Activation requires the UreD and UreF proteins, each encoded by two paralogues. UreG, a third essential activation protein, is encoded by the single-copy Eu3, and eu3 mutants lack activity of both urease isozymes. eu2 has the same urease-negative phenotype, consistent with Eu2 being a single-copy gene, possibly encoding a Ni carrier. Unexpectedly, two eu2 alleles co-segregated with missense mutations in the chromosome 2 UreF paralogue (Ch02UreF), suggesting lack of expression/function of Ch14UreF. However, Ch02UreF and Ch14UreF transcripts accumulate at the same level. Further, it had been shown that expression of the Ch14UreF ORF complemented a fungal ureF mutant. A third, nonsense (Q2*) allelic mutant, eu2-c, exhibited 5- to 10-fold more residual urease activity than missense eu2-a or eu2-b, though eu2-c should lack all Ch02UreF protein. It is hypothesized that low-level activation by Ch14UreF is ‘spoiled’ by the altered missense Ch02UreF proteins (‘epistatic dominant-negative’). In agreement with active ‘spoiling’ by eu2-b-encoded Ch02UreF (G31D), eu2-b/eu2-c heterozygotes had less than half the urease activity of eu2-c/eu2-c siblings. Ch02UreF (G31D) could spoil activation by Chr14UreF because of higher affinity for the activation complex, or because Ch02UreF (G31D) is more abundant than Ch14UreF. Here, the latter is favoured, consistent with a reported in-frame AUG in the 5' leader of Chr14UreF transcript. Translational inhibition could represent a form of ‘functional divergence’ of duplicated genes. PMID:21430294

  14. Molecular investigation of virulence factors of Brucella melitensis and Brucella abortus strains isolated from clinical and non-clinical samples.

    PubMed

    Mirnejad, Reza; Jazi, Faramarz Masjedian; Mostafaei, Shayan; Sedighi, Mansour

    2017-08-01

    Brucella is zoonotic pathogen that induces abortion and sterility in domestic mammals and chronic infections in humans called Malta fever. It is a facultative intracellular potential pathogen with high infectivity. The virulence of Brucella is dependent upon its potential virulence factors such as enzymes and cell envelope associated virulence genes. The aim of this study was to investigate the Brucella virulence factors among strains isolated from humans and animals in different parts of Iran. Seventy eight strains of Brucella species isolated from suspected human and animal cases from several provinces of Iran during 2015-2016 and identified by phenotypic and molecular methods. The multiplex-PCR (M-PCR) assay was performed in order to detect the ure, wbkA, omp19, mviN, manA and perA genes by using gene specific primers. Out of 78 isolates of Brucella spp., 57 (73%) and 21 (27%) isolates were detected as B. melitensis and B. abortus, respectively, by molecular method. The relative frequency of virulence genes ure, wbkA, omp19, mviN, manA and perA were 74.4%, 89.7%, 93.6%, 94.9%, 100% and 92.3%, respectively. Our results indicate that the most of Brucella strains isolated from this region possess high percent of virulence factor genes (ure, wbkA, omp19, mviN, manA and perA) in their genome. So, each step of infection can be mediated by a number of virulence factors and each strain may have a unique combination of these factors that affected the rate of bacterial pathogenesis. Copyright © 2017 Elsevier Ltd. All rights reserved.

  15. The 26S Proteasome Degrades the Soluble but Not the Fibrillar Form of the Yeast Prion Ure2p In Vitro

    PubMed Central

    Wang, Kai; Redeker, Virginie; Madiona, Karine; Melki, Ronald; Kabani, Mehdi

    2015-01-01

    Yeast prions are self-perpetuating protein aggregates that cause heritable and transmissible phenotypic traits. Among these, [PSI +] and [URE3] stand out as the most studied yeast prions, and result from the self-assembly of the translation terminator Sup35p and the nitrogen catabolism regulator Ure2p, respectively, into insoluble fibrillar aggregates. Protein quality control systems are well known to govern the formation, propagation and transmission of these prions. However, little is known about the implication of the cellular proteolytic machineries in their turnover. We previously showed that the 26S proteasome degrades both the soluble and fibrillar forms of Sup35p and affects [PSI +] propagation. Here, we show that soluble native Ure2p is degraded by the proteasome in an ubiquitin-independent manner. Proteasomal degradation of Ure2p yields amyloidogenic N-terminal peptides and a C-terminal resistant fragment. In contrast to Sup35p, fibrillar Ure2p resists proteasomal degradation. Thus, structural variability within prions may dictate their ability to be degraded by the cellular proteolytic systems. PMID:26115123

  16. The assembly of the plant urease activation complex and the essential role of the urease accessory protein G (UreG) in delivery of nickel to urease.

    PubMed

    Myrach, Till; Zhu, Anting; Witte, Claus-Peter

    2017-09-01

    Urease is a ubiquitous nickel metalloenzyme. In plants, its activation requires three urease accessory proteins (UAPs), UreD, UreF, and UreG. In bacteria, the UAPs interact with urease and facilitate activation, which involves the channeling of two nickel ions into the active site. So far this process has not been investigated in eukaryotes. Using affinity pulldowns of Strep-tagged UAPs from Arabidopsis and rice transiently expressed in planta , we demonstrate that a urease-UreD-UreF-UreG complex exists in plants and show its stepwise assembly. UreG is crucial for nickel delivery because UreG-dependent urease activation in vitro was observed only with UreG obtained from nickel-sufficient plants. This activation competence could not be generated in vitro by incubation of UreG with nickel, bicarbonate, and GTP. Compared with their bacterial orthologs, plant UreGs possess an N-terminal extension containing a His- and Asp/Glu-rich hypervariable region followed by a highly conserved sequence comprising two potential H X H metal-binding sites. Complementing the ureG-1 mutant of Arabidopsis with N-terminal deletion variants of UreG demonstrated that the hypervariable region has a minor impact on activation efficiency, whereas the conserved region up to the first H X H motif is highly beneficial and up to the second H X H motif strictly required for activation. We also show that urease reaches its full activity several days after nickel becomes available in the leaves, indicating that urease activation is limited by nickel accessibility in vivo Our data uncover the crucial role of UreG for nickel delivery during eukaryotic urease activation, inciting further investigations of the details of this process. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

  17. Differential diagnosis between complete mole and hydropic abortus by deoxyribonucleic acid fingerprints.

    PubMed

    Nobunaga, T; Azuma, C; Kimura, T; Tokugawa, Y; Takemura, M; Kamiura, S; Saji, F; Tanizawa, O

    1990-08-01

    We used a new method of deoxyribonucleic acid fingerprint analysis to obtain the differential diagnosis between complete mole and hydropic abortus. This method with a deoxyribonucleic acid minisatellite probe requires only a small amount of tissue sample and peripheral blood, and presents individual specific restriction fragment length polymorphisms (deoxyribonucleic acid "fingerprints") by simultaneous detection of many hypervariable regions (minisatellite regions) widely dispersed in the human genome. Southern blot hybridization showed that in cases of complete mole, all polymorphic fragments were exclusively inherited from the father. Some of the polymorphic bands of paternal deoxyribonucleic acid were not observed in molar deoxyribonucleic acid. However, in the hydropic abortus, the polymorphic fragments could be traced back to its parent. These results indicate that deoxyribonucleic acid fingerprints could distinguish the abnormal fertilization of complete mole (androgenesis) from the normal fertilization of hydropic abortus by identifying the difference in genetic variations between complete mole and hydropic abortus at the deoxyribonucleic acid level.

  18. 9 CFR 113.65 - Brucella Abortus Vaccine.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 9 Animals and Animal Products 1 2012-01-01 2012-01-01 false Brucella Abortus Vaccine. 113.65... Bacterial Vaccines § 113.65 Brucella Abortus Vaccine. Brucella Abortus Vaccine shall be prepared as a desiccated live culture bacterial vaccine from smooth colonial forms of the Brucella abortus organism...

  19. 9 CFR 113.65 - Brucella Abortus Vaccine.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 9 Animals and Animal Products 1 2011-01-01 2011-01-01 false Brucella Abortus Vaccine. 113.65... Bacterial Vaccines § 113.65 Brucella Abortus Vaccine. Brucella Abortus Vaccine shall be prepared as a desiccated live culture bacterial vaccine from smooth colonial forms of the Brucella abortus organism...

  20. 9 CFR 113.65 - Brucella Abortus Vaccine.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 9 Animals and Animal Products 1 2014-01-01 2014-01-01 false Brucella Abortus Vaccine. 113.65... Bacterial Vaccines § 113.65 Brucella Abortus Vaccine. Brucella Abortus Vaccine shall be prepared as a desiccated live culture bacterial vaccine from smooth colonial forms of the Brucella abortus organism...

  1. 9 CFR 113.65 - Brucella Abortus Vaccine.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 9 Animals and Animal Products 1 2013-01-01 2013-01-01 false Brucella Abortus Vaccine. 113.65... Bacterial Vaccines § 113.65 Brucella Abortus Vaccine. Brucella Abortus Vaccine shall be prepared as a desiccated live culture bacterial vaccine from smooth colonial forms of the Brucella abortus organism...

  2. 9 CFR 113.65 - Brucella Abortus Vaccine.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 9 Animals and Animal Products 1 2010-01-01 2010-01-01 false Brucella Abortus Vaccine. 113.65... Bacterial Vaccines § 113.65 Brucella Abortus Vaccine. Brucella Abortus Vaccine shall be prepared as a desiccated live culture bacterial vaccine from smooth colonial forms of the Brucella abortus organism...

  3. An outbreak of Brucella abortus biovar 2 in Canadian cattle

    PubMed Central

    Forbes, Lorry B.; Steele, Thomas B.

    1989-01-01

    An outbreak of brucellosis caused by Brucella abortus biovar 2 was identified in cattle in Alberta in December 1986. This was the only clinical infection discovered since the national cattle herd was declared brucellosisfree in 1985. It was the first report of B. abortus biovar 2 in Canadian cattle. The outbreak, involving three herds containing purebred Hereford cattle, was spread by the private treaty sale of untested cattle, and was identified following investigation of an abortion. The source of infection for the outbreak was not established, but several possibilities were identified including infected herds present in the area during the mid-1970's, latent infection originating in a Saskatchewan herd during the early 1960's, American cattle imported during the early 1970's, and brucellosis-infected bison in Wood Buffalo National Park. The containment and elimination of this nidus of infection appears to have been successful, and the national cattle herd at the time of writing is free of the disease. PMID:17423457

  4. [Detection of a clonal complex with Brucella abortus biovar 2 genotype as founder in B. abortus isolates from Argentina].

    PubMed

    Hollender, Daiana; Conde, Sandra B; Salustio, Eduardo; Samartino, Luis E

    2013-01-01

    Brucella abortus is the causative agent of bovine brucellosis, a worldwide zoonosis. Up to date, eight biovars of B. abortus have been described. In Argentina, biovar 1 is the most frequently isolated. However, biovar 2, which is more pathogenic than biovar 1, is also found. Molecular methods for subtyping isolates are necessary for allowing epidemiological surveillance and control of eradication programs. Due to the genetic homogeneity of the genus Brucella, the development of molecular typing tools has been difficult. The publication of microorganism genomes facilitates the design of this approach. The aim of this work was to employ a Multiple Locus VNTR Analysis (MLVA) scheme for strains from Argentina isolated in our laboratory. From the 56 isolates analyzed, 47 different genotypic profiles were obtained. All the strains typed as biovar 2 showed the same profile. This scheme allowed assigning each isolate to the biovar it belongs to. All the genotypes were related using the goeBURST analysis and biovar 2 was proposed as founder. Copyright © 2013 Asociación Argentina de Microbiología. Publicado por Elsevier España. All rights reserved.

  5. Variables Associated with Infections of Cattle by Brucella abortus., Leptospira spp. and Neospora spp. in Amazon Region in Brazil.

    PubMed

    Chiebao, D P; Valadas, S Y O B; Minervino, A H H; Castro, V; Romaldini, A H C N; Calhau, A S; De Souza, R A B; Gennari, S M; Keid, L B; Soares, R M

    2015-10-01

    The frequency of Neospora spp., Leptospira spp. and Brucella abortus infections in adult cattle was determined in herds of the State of Pará, Brazil, which is an important region for cattle production located in the Amazon region. A total of 3466 adult female cattle from 176 herds were tested, leading to a frequency of seropositive animals of 14.7%, 3.7% and 65.5% and a herd positivity of 87.4%, 41.3% and 98.8% for infections caused by Neospora spp., B. abortus and Leptospira spp., respectively. The five most frequently diagnosed serologic responses to Leptospira spp. were those against serovars hardjo, wolfii, grippotyphosa, hebdomadis and shermani. The following associations were found: practice of artificial insemination, large farm size, large herd size, large number of dogs and high number of total abortions per year with the presence of antibodies against serovar hardjo; positive results to serovar grippotyphosa with the presence of dogs; inappropriate disposal of aborted foetuses with positivity to serovar hebdomadis. Serovar grippotyphosa was also associated with number of episodes of abortions. Neospora spp. positive herds were associated with episodes of abortion and B. abortus infection with the disposal of dead animals and aborted foetuses on pastures and with the use of artificial insemination. In conclusion, the high frequency of brucellosis, leptospirosis and neosporosis in the region may be a consequence of social, natural and raising conditions as: (i) climate conditions that favour the survival and spread of pathogens in the environment; (ii) farms located in regions bordering forest areas; (iii) farms in areas of difficult access to the veterinary service; (iv) extensive beef herds raised at pastures with different age and productive groups inter-mingled; and (v) minimal concerns regarding hygiene practices and disease prevention measures. © 2013 Blackwell Verlag GmbH.

  6. Molecular characterization of Brucella abortus chromosome II recombination.

    PubMed

    Tsoktouridis, Georgios; Merz, Christian A; Manning, Simon P; Giovagnoli-Kurtz, Renée; Williams, Leanne E; Mujer, Cesar V; Hagius, Sue; Elzer, Philip; Redkar, Rajendra J; Patra, Guy; DelVecchio, Vito G

    2003-10-01

    Large-scale genomic rearrangements including inversions, deletions, and duplications are significant in bacterial evolution. The recently completed Brucella melitensis 16M and Brucella suis 1330 genomes have facilitated the investigation of such events in the Brucella spp. Suppressive subtractive hybridization (SSH) was employed in identifying genomic differences between B. melitensis 16M and Brucella abortus 2308. Analysis of 45 SSH clones revealed several deletions on chromosomes of B. abortus and B. melitensis that encoded proteins of various metabolic pathways. A 640-kb inversion on chromosome II of B. abortus has been reported previously (S. Michaux Charachon, G. Bourg, E. Jumas Bilak, P. Guigue Talet, A. Allardet Servent, D. O'Callaghan, and M. Ramuz, J. Bacteriol. 179:3244-3249, 1997) and is further described in this study. One end of the inverted region is located on a deleted TATGC site between open reading frames BMEII0292 and BMEII0293. The other end inserted at a GTGTC site of the cyclic-di-GMP phosphodiesterase A (PDEA) gene (BMEII1009), dividing PDEA into two unequal DNA segments of 160 and 977 bp. As a consequence of inversion, the 160-bp segment that encodes the N-terminal region of PDEA was relocated at the opposite end of the inverted chromosomal region. The splitting of the PDEA gene most likely inactivated the function of this enzyme. A recombination mechanism responsible for this inversion is proposed.

  7. Molecular Characterization of Brucella abortus Chromosome II Recombination

    PubMed Central

    Tsoktouridis, Georgios; Merz, Christian A.; Manning, Simon P.; Giovagnoli-Kurtz, Renée; Williams, Leanne E.; Mujer, Cesar V.; Hagius, Sue; Elzer, Philip; Redkar, Rajendra J.; Patra, Guy; DelVecchio, Vito G.

    2003-01-01

    Large-scale genomic rearrangements including inversions, deletions, and duplications are significant in bacterial evolution. The recently completed Brucella melitensis 16M and Brucella suis 1330 genomes have facilitated the investigation of such events in the Brucella spp. Suppressive subtractive hybridization (SSH) was employed in identifying genomic differences between B. melitensis 16M and Brucella abortus 2308. Analysis of 45 SSH clones revealed several deletions on chromosomes of B. abortus and B. melitensis that encoded proteins of various metabolic pathways. A 640-kb inversion on chromosome II of B. abortus has been reported previously (S. Michaux Charachon, G. Bourg, E. Jumas Bilak, P. Guigue Talet, A. Allardet Servent, D. O'Callaghan, and M. Ramuz, J. Bacteriol. 179:3244-3249, 1997) and is further described in this study. One end of the inverted region is located on a deleted TATGC site between open reading frames BMEII0292 and BMEII0293. The other end inserted at a GTGTC site of the cyclic-di-GMP phosphodiesterase A (PDEA) gene (BMEII1009), dividing PDEA into two unequal DNA segments of 160 and 977 bp. As a consequence of inversion, the 160-bp segment that encodes the N-terminal region of PDEA was relocated at the opposite end of the inverted chromosomal region. The splitting of the PDEA gene most likely inactivated the function of this enzyme. A recombination mechanism responsible for this inversion is proposed. PMID:14526025

  8. Swa2, the yeast homolog of mammalian auxilin, is specifically required for the propagation of the prion variant [URE 3‐1

    PubMed Central

    Troisi, Elizabeth M.; Rockman, Michael E.; Nguyen, Phil P.; Oliver, Emily E.

    2015-01-01

    Summary Yeast prions require a core set of chaperone proteins including Sis1, Hsp70 and Hsp104 to generate new amyloid templates for stable propagation, yet emerging studies indicate that propagation of some prions requires additional chaperone activities, demonstrating chaperone specificity beyond the common amyloid requirements. To comprehensively assess such prion‐specific requirements for the propagation of the [URE 3] prion variant [URE 3‐1], we screened 12 yeast cytosolic J‐proteins, and here we report a novel role for the J‐protein Swa2/Aux1. Swa2 is the sole yeast homolog of the mammalian protein auxilin, which, like Swa2, functions in vesicle‐mediated endocytosis by disassembling the structural lattice formed by the protein clathrin. We found that, in addition to Sis1, [URE 3‐1] is specifically dependent upon Swa2, but not on any of the 11 other J‐proteins. Further, we show that [URE 3‐1] propagation requires both a functional J‐domain and the tetratricopeptide repeat (TPR) domain, but surprisingly does not require Swa2‐clathrin binding. Because the J‐domain of Swa2 can be replaced with the J‐domains of other proteins, our data strongly suggest that prion‐chaperone specificity arises from the Swa2 TPR domain and supports a model where Swa2 acts through Hsp70, most likely to provide additional access points for Hsp104 to promote prion template generation. PMID:26031938

  9. Undergraduate Research Experience in Ocean/Marine Science (URE-OMS)

    DTIC Science & Technology

    2003-09-30

    The URE-Ocean/Marine Science program supports active research participation by undergraduate students in remote sensing and GIS. The program is based on a model for undergraduate research programs supported by the National Science Foundation . URE project features mentors, research projects, and professional development opportunities. It is the long-term goal

  10. Prevalence and molecular identification of Chlamydia abortus in commercial dairy goat farms in a hot region in Mexico.

    PubMed

    Campos-Hernández, Eleuterio; Vázquez-Chagoyán, Juan Carlos; Salem, Abdelfattah Z M; Saltijeral-Oaxaca, Jorge Antonio; Escalante-Ochoa, Cristina; López-Heydeck, Sandra M; de Oca-Jiménez, Roberto Montes

    2014-08-01

    The aim of this study was to determine the seroprevalence and presence of Chlamydia abortus in Saanen breed female goats from commercial dairy goat farms under intensive production in the municipality of Guanajuato, Mexico. Sera were collected to determine the prevalence of anti-C. abortus IgG antibodies using recombinant enzyme-linked immunosorbent assay (rELISA) and cell culture. Polymerase chain reaction (PCR) was used to prove the presence of the pathogen in swab samples collected from the vagina and rectum of selected animals. Additionally, foetal tissue samples from a sudden abortion were collected. C. abortus prevalence in female goats of commercial milking farms sampled in Guanajuato, Mexico, was 4.87% (n = 246). Seropositive animals were found in six out of nine (66.6%) dairy goat farms sampled, and prevalence among animals in individual farms ranged between 3.44 and 13.51%. C. abortus was detected using PCR in spleen tissue from the aborted foetus. PCR-based detection, as well as isolation from vaginal and rectal swabs, was not possible in the present study. Isolation through cell culture was also unsuccessful from aborted foetal tissue samples. In conclusion, the results from rELISA and PCR show that C. abortus is present in dairy goat farms in the state of Guanajuato, Mexico.

  11. Molecular Cloning and Characterization of cgs, the Brucella abortus Cyclic β(1-2) Glucan Synthetase Gene: Genetic Complementation of Rhizobium meliloti ndvB and Agrobacterium tumefaciens chvB Mutants

    PubMed Central

    Iñón de Iannino, Nora; Briones, Gabriel; Tolmasky, Marcelo; Ugalde, Rodolfo A.

    1998-01-01

    The animal pathogen Brucella abortus contains a gene, cgs, that complemented a Rhizobium meliloti nodule development (ndvB) mutant and an Agrobacterium tumefaciens chromosomal virulence (chvB) mutant. The complemented strains recovered the synthesis of cyclic β(1-2) glucan, motility, virulence in A. tumefaciens, and nitrogen fixation in R. meliloti; all traits were strictly associated with the presence of an active cyclic β(1-2) glucan synthetase protein in the membranes. Nucleotide sequencing revealed the presence in B. abortus of an 8.49-kb open reading frame coding for a predicted membrane protein of 2,831 amino acids (316.2 kDa) and with 51% identity to R. meliloti NdvB. Four regions of the B. abortus protein spanning amino acids 520 to 800, 1025 to 1124, 1284 to 1526, and 2400 to 2660 displayed similarities of higher than 80% with R. meliloti NdvB. Tn3-HoHo1 mutagenesis showed that the C-terminal 825 amino acids of the Brucella protein, although highly conserved in Rhizobium, are not necessary for cyclic β(1-2) glucan synthesis. Confirmation of the identity of this protein as B. abortus cyclic β(1-2) glucan synthetase was done by the construction of a B. abortus Tn3-HoHo1 insertion mutant that does not form cyclic β(1-2) glucan and lacks the 316.2-kDa membrane protein. The recovery of this mutant from the spleens of inoculated mice was decreased by 3 orders of magnitude compared with that of the parental strain; this result suggests that cyclic β(1-2) glucan may be a virulence factor in Brucella infection. PMID:9721274

  12. Brucella abortus Cyclic β-1,2-Glucan Mutants Have Reduced Virulence in Mice and Are Defective in Intracellular Replication in HeLa Cells

    PubMed Central

    Briones, Gabriel; Iñón de Iannino, Nora; Roset, Mara; Vigliocco, Ana; Paulo, Patricia Silva; Ugalde, Rodolfo A.

    2001-01-01

    Null cyclic β-1,2-glucan synthetase mutants (cgs mutants) were obtained from Brucella abortus virulent strain 2308 and from B. abortus attenuated vaccinal strain S19. Both mutants show greater sensitivity to surfactants like deoxycholic acid, sodium dodecyl sulfate, and Zwittergent than the parental strains, suggesting cell surface alterations. Although not to the same extent, both mutants display reduced virulence in mice and defective intracellular multiplication in HeLa cells. The B. abortus S19 cgs mutant was completely cleared from the spleens of mice after 4 weeks, while the 2308 mutant showed a 1.5-log reduction of the number of brucellae isolated from the spleens after 12 weeks. These results suggest that cyclic β-1,2-glucan plays an important role in the residual virulence of the attenuated B. abortus S19 strain. Although the cgs mutant was cleared from the spleens earlier than the wild-type parental strain (B. abortus S19) and produced less inflammatory response, its ability to confer protection against the virulent strain B. abortus 2308 was fully retained. Equivalent levels of induction of spleen gamma interferon mRNA and anti-lipopolysaccharide (LPS) of immunoglobulin G2a (IgG2a) subtype antibodies were observed in mice injected with B. abortus S19 or the cgs mutant. However, the titer of anti-LPS antibodies of the IgG1 subtype induced by the cgs mutant was lower than that observed with the parental S19 strain, thus suggesting that the cgs mutant induces a relatively exclusive Th1 response. PMID:11401996

  13. Seroprevalence and Risk Factors of Chlamydia abortus Infection in Tibetan Sheep in Gansu Province, Northwest China

    PubMed Central

    Qin, Si-Yuan; Yin, Ming-Yang; Cong, Wei; Zhou, Dong-Hui; Zhang, Xiao-Xuan; Zhao, Quan; Zhu, Xing-Quan; Zhou, Ji-Zhang; Qian, Ai-Dong

    2014-01-01

    Chlamydia abortus, an important pathogen in a variety of animals, is associated with abortion in sheep. In the present study, 1732 blood samples, collected from Tibetan sheep between June 2013 and April 2014, were examined by the indirect hemagglutination (IHA) test, aiming to evaluate the seroprevalence and risk factors of C. abortus infection in Tibetan sheep. 323 of 1732 (18.65%) samples were seropositive for C. abortus antibodies at the cut-off of 1 : 16. A multivariate logistic regression analysis was used to evaluate the risk factors associated with seroprevalence, which could provide foundation to prevent and control C. abortus infection in Tibetan sheep. Gender of Tibetan sheep was left out of the final model because it is not significant in the logistic regression analysis (P > 0.05). Region, season, and age were considered as major risk factors associated with C. abortus infection in Tibetan sheep. Our study revealed a widespread and high prevalence of C. abortus infection in Tibetan sheep in Gansu province, northwest China, with higher exposure risk in different seasons and ages and distinct geographical distribution. PMID:25401129

  14. The transport kinetics and selectivity of HpUreI, the urea channel from Helicobacter pylori†

    PubMed Central

    Gray, Lawrence R; Gu, Sean X; Quick, Matthias; Khademi, Shahram

    2017-01-01

    Helicobacter pylori’s unique ability to colonize and survive in the acidic environment of the stomach is critically dependent on uptake of urea through the urea channel, HpUreI. Hence, HpUreI may represent a promising target for the development of specific drugs against this human pathogen. To obtain insight into the structure/function relationship of this channel, we have developed conditions for the high-yield expression and purification of stable recombinant HpUreI that allowed its detailed kinetic characterization in solubilized form and reconstituted into liposomes. Detergent-solubilized HpUreI forms homo-trimer, as determined by chemical cross-linking. Urea dissociation kinetics of purified HpUreI were determined by means of the scintillation proximity assay (SPA), whereas urea efflux was measured in HpUreI-containing proteoliposomes using stopped-flow spectrometry to determine the kinetics and selectivity of the urea channel. The kinetic analyses revealed that urea conduction in HpUreI is pH sensitive and saturable with a half-saturation concentration (or K0.5) of ~163 mM. Binding of urea by HpUreI was increased at lower pH; however, the apparent affinity of urea binding (~150 mM) was not significantly pH dependent. The solute selectivity analysis indicated that HpUreI is highly selective for urea and hydroxyurea. Removing either amino group of urea molecules diminishes their permeability through HpUreI. Similar to urea conduction, water diffusion through HpUreI is pH-dependent with low water permeability at neutral pH. PMID:21877689

  15. Detection of antibodies against Chlamydophila abortus in Costa Rican sheep flocks

    PubMed Central

    Villagra-Blanco, R.; Dolz, G.; Montero-Caballero, D.; Romero-Zúñiga, J.J.

    2015-01-01

    A total of 359 sheep samples from 15 flocks were analyzed for the presence of antibodies against Chlamydophila abortus using a commercial Enzyme linked Immunosorbent Assay (ELISA). Antibodies were detected in 19 (5.29%) sheep from 12 (80%) flocks. Seropositive animals were found in all analyzed regions (Central, Chorotega, Atlantic Huetar, North Huetar and Central Pacific) determining prevalence between 0.28% and 4.4%, and intra-flock positivity between 3.7% and 25.0%. The survey revealed two risk factors associated with seropositivity; introducing animals (males and females), embryos, or semen from other farms or from abroad without any sanitary certification, and flocks not having quarantine areas or separated boxes for diseased animals. No clinical signs of disease were observed in positive seroreactors. C. abortus seems to be present in Costa Rica in a very low prevalence in sheep flocks. Further studies, to isolate the bacteria are required. Finally, implementation of control measures to prevent the spread of C. abortus is recommended. PMID:26623377

  16. Non-thiolate ligation of nickel by nucleotide-free UreG of Klebsiella aerogenes

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Martin-Diaconescu, Vlad; Joseph, Crisjoe A.; Boer, Jodi L.

    Nickel-dependent ureases are activated by a multiprotein complex that includes the GTPase UreG. Prior studies showed that nucleotide-free UreG from Klebsiella aerogenes is monomeric and binds one nickel or zinc ion with near-equivalent affinity using an undefined binding site, whereas nucleotide-free UreG from Helicobacter pylori selectively binds one zinc ion per dimer via a universally conserved Cys-Pro-His motif in each protomer. Iodoacetamide-treated K. aerogenes UreG was nearly unaffected in nickel binding compared to non-treated sample, suggesting the absence of thiolate ligands to the metal. X-ray absorption spectroscopy of nickel-bound UreG showed the metal possessed four-coordinate geometry with all O/N donormore » ligands including one imidazole, thus confirming the absence of thiolate ligation. The nickel site in Strep-tag II-modified protein possessed six-coordinate geometry, again with all O/N donor ligands, but now including two or three imidazoles. An identical site was noted for the Strep-tag II-modified H74A variant, substituted in the Cys-Pro-His motif, ruling out coordination by this His residue. These results are consistent with metal binding to both His6 and a His residue of the fusion peptide in Strep-tagged K. aerogenes UreG. We conclude that the nickel- and zinc-binding site in nucleotide-free K. aerogenes UreG is distinct from that of nucleotide-free H. pylori UreG and does not involve the Cys-Pro-His motif. Further, we show the Strep-tag II can perturb metal coordination of this protein.« less

  17. Seroprevalence and risk factors of Chlamydia abortus infection in free-ranging white yaks in China.

    PubMed

    Qin, Si-Yuan; Huang, Si-Yang; Yin, Ming-Yang; Tan, Qi-Dong; Liu, Guang-Xue; Zhou, Dong-Hui; Zhu, Xing-Quan; Zhou, Ji-Zhang; Qian, Ai-Dong

    2015-01-20

    Chlamydia is gram-negative obligate bacteria which causes a wide variety of diseases in humans and animals. To date, there are a few reports about the seroprevalence of Chlamydia and the risk factors associated with Chlamydia infection in yaks in the world. In this study, 974 blood samples were collected from white yaks (Bos grunniens) in Tianzhu Tibetan Autonomous County, Gansu province, northwest China from June 2013 to April 2014. Antibodies against Chlamydia abortus were examined by the indirect hemagglutination (IHA) test, and 158 of 974 (16.22%) white yaks were seropositive for C. abortus antibodies at the cut-off of 1:16. The risk factors associated with seroprevalence were evaluated by a multivariate logistic regression analysis. Region, gender and age of white yak were left out of the final model, due to its insignificance in the logistic regression analysis (P > 0.05). However, season was considered as a major risk factor associated with C. abortus infection in white yaks. To our knowledge, this is the first survey of C. abortus seroprevalence in white yaks in China, which extends the host range for C. abortus and has important implications for public health and the local Tibetan economy.

  18. Archaeal amoA and ureC genes and their transcriptional activity in the Arctic Ocean

    NASA Astrophysics Data System (ADS)

    Pedneault, Estelle; Galand, Pierre E.; Potvin, Marianne; Tremblay, Jean-Éric; Lovejoy, Connie

    2014-04-01

    Thaumarchaeota and the gene encoding for a subunit of ammonia monooxygenase (amoA) are ubiquitous in Polar Seas, and some Thaumarchaeota also have a gene coding for ureC, diagnostic for urease. Using quantitative PCR we investigated the occurrence of genes and transcripts of ureC and amoA in Arctic samples from winter, spring and summer. AmoA genes, ureC genes and amoA transcripts were always present, but ureC transcripts were rarely detected. Over a 48 h light manipulation experiment amoA transcripts persisted under light and dark conditions, but not ureC transcripts. In addition, maxima for amoA transcript were nearer the surface compared to amoA genes. Clone libraries using DNA template recovered shallow and deep amoA clades but only the shallow clade was recovered from cDNA (from RNA). These results imply environmental control of amoA expression with direct or indirect light effects, and rare ureC expression despite its widespread occurrence in the Arctic Ocean.

  19. A Homologue of an Operon Required for DNA Transfer in Agrobacterium Is Required in Brucella abortus for Virulence and Intracellular Multiplication

    PubMed Central

    Sieira, Rodrigo; Comerci, Diego J.; Sánchez, Daniel O.; Ugalde, Rodolfo A.

    2000-01-01

    As part of a Brucella abortus 2308 genome project carried out in our laboratory, we identified, cloned, and sequenced a genomic DNA fragment containing a locus (virB) highly homologous to bacterial type IV secretion systems. The B. abortus virB locus is a collinear arrangement of 13 open reading frames (ORFs). Between virB1 and virB2 and downstream of ORF12, two degenerated, palindromic repeat sequences characteristic of Brucella intergenic regions were found. Gene reporter studies demonstrated that the B. abortus virB locus constitutes an operon transcribed from virB1 which is turned on during the stationary phase of growth. A B. abortus polar virB1 mutant failed to replicate in HeLa cells, indicating that the virB operon plays a critical role in intracellular multiplication. Mutants with polar and nonpolar mutations introduced in virB10 showed different behaviors in mice and in the HeLa cell infection assay, suggesting that virB10 per se is necessary for the correct function of this type IV secretion apparatus. Mouse infection assays demonstrated that the virB operon constitutes a major determinant of B. abortus virulence. It is suggested that putative effector molecules secreted by this type IV secretion system determine routing of B. abortus to an endoplasmic reticulum-related replication compartment. PMID:10940027

  20. Proteomic Profile of Brucella abortus-Infected Bovine Chorioallantoic Membrane Explants

    PubMed Central

    Mol, Juliana P. S.; Pires, Simone F.; Chapeaurouge, Alexander D.; Perales, Jonas; Santos, Renato L.; Andrade, Hélida M.; Lage, Andrey P.

    2016-01-01

    Brucella abortus is the etiological agent of bovine brucellosis, a zoonotic disease that causes significant economic losses worldwide. The differential proteomic profile of bovine chorioallantoic membrane (CAM) explants at early stages of infection with B. abortus (0.5, 2, 4, and 8 h) was determined. Analysis of CAM explants at 0.5 and 4 h showed the highest differences between uninfected and infected CAM explants, and therefore were used for the Differential Gel Electrophoresis (DIGE). A total of 103 spots were present in only one experimental group and were selected for identification by mass spectrometry (MALDI/ToF-ToF). Proteins only identified in extracts of CAM explants infected with B. abortus were related to recognition of PAMPs by TLR, production of reactive oxygen species, intracellular trafficking, and inflammation. PMID:27104343

  1. Humoral immune response against lipopolysaccharide and cytoplasmic proteins of Brucella abortus in cattle vaccinated with B. abortus S19 or experimentally infected with Yersinia enterocolitica serotype 0:9.

    PubMed Central

    Baldi, P C; Giambartolomei, G H; Goldbaum, F A; Abdón, L F; Velikovsky, C A; Kittelberger, R; Fossati, C A

    1996-01-01

    The humoral immune responses against three different antigens of Brucella abortus were monitored by enzyme-linked immunosorbent assay in cattle vaccinated with B. abortus S19 or experimentally infected with Yersinia enterocolitica serotype 0:9. Immunoglobulin G (IgG) and IgM responses against (i) B. abortus lipopolysaccharide (LPS), (ii) total cytoplasmic proteins depleted of LPS (LPS-free CYT), and (iii) B. abortus 18-kDa cytoplasmic protein were measured. Vaccinated animals and Yersinia-infected animals developed high anti-LPS IgM and IgG titers, which overlapped with those obtained with sera from B. abortus 544-infected animals used as positive controls. In contrast, only a slight or negative IgG and IgM response against LPS-free CYT and the 18-kDa protein was detected in vaccinated or Yersinia-infected cattle, although its levels were always significantly lower than those of B. abortus 544-infected animals. These data indicate that cytoplasmic proteins of B. abortus could be useful for the differential diagnosis of bovine brucellosis. PMID:8807216

  2. Immunogenic proteins of Brucella abortus to minimize cross reactions in brucellosis diagnosis.

    PubMed

    Ko, Kyung Yuk; Kim, Jong-Wan; Her, Moon; Kang, Sung-Il; Jung, Suk Chan; Cho, Dong Hee; Kim, Ji-Yeon

    2012-05-04

    To overcome the limitations of serological diagnosis, including false positive reactions caused by other pathogens, specific antigens for diagnosis of brucellosis other than LPS have been required. The present study was conducted to separate and identify immuno-dominant insoluble proteins of Brucella abortus against the antisera of cattle infected with B. abortus, or/and Yersinia enterocolitica, or the sera of non-infected cattle. After separating insoluble proteins of B. abortus by two dimensional electrophoresis (2-DE), their immuno-reactivity was determined by western blotting. A portion of the immunogenic spots against the positive antisera of B. abortus that have the potential for use as specific antigens were identified by MS/MS analysis. Overall, 18 immunogenic insoluble proteins of B. abortus 1119-3 showed immuno-reactivity against only the positive antisera of B. abortus, but failed to have immunogenicity toward both the positive sera of Y. enterocolitica and the negative sera of B. abortus. Identification of these proteins revealed the following: F0F1 ATP synthase subunit β, solute-binding family 5 protein, 28 kDa OMP, Leu/Ile/Val-binding family protein, Histidinol dehyddrogenase, Hypothetical protein, Twin-arginine translocation pathway signal sequence domain-containing protein, Dihydroorotase, Serine protease family protein, β-hydroxyacyl-(acyl-carrier-protein) dehydratase FabA, Short-chain dehydrogenase-/reductase carbonic anhydrase, Orinithine carbamoyltransferase, Leucyl aminopeptidase, Cold shock DNA-binding domain-containing protein, Cu/Zn superoxide dismutase, and Methionine aminopeptidase. The 18 immunogenic proteins separated in the present study can be considered candidate antigens to minimize cross reaction in the diagnosis of brucellosis and useful sources for Brucella vaccine development. Copyright © 2011 Elsevier B.V. All rights reserved.

  3. TLR9 is required for MAPK/NF-κB activation but does not cooperate with TLR2 or TLR6 to induce host resistance to Brucella abortus.

    PubMed

    Gomes, Marco Túlio; Campos, Priscila Carneiro; Pereira, Guilherme de Sousa; Bartholomeu, Daniella Castanheira; Splitter, Gary; Oliveira, Sergio Costa

    2016-05-01

    Brucella abortus is a Gram-negative intracellular bacterial pathogen that causes a zoonosis of worldwide occurrence, leading to undulant fever in humans and abortion in domestic animals. B. abortus is recognized by several pattern-recognition receptors triggering pathways during the host innate immune response. Therefore, here, we determined the cooperative role of TLR9 with TLR2 or TLR6 receptors in sensing Brucella Furthermore, we deciphered the host innate immune response against B. abortus or its DNA, emphasizing the role of TLR9-MAPK/NF-κB signaling pathways in the production of proinflammatory cytokines. TLR9 is required for the initial host control of B. abortus, but this TLR was dispensable after 6 wk of infection. The susceptibility of TLR9(-/-)-infected animals to Brucella paralleled with lower levels of IFN-γ produced by mouse splenocytes stimulated with this pathogen compared with wild-type cells. However, no apparent cooperative interplay was observed between TLR2-TLR9 or TLR6-TLR9 receptors to control infection. Moreover, B. abortus or its DNA induced activation of MAPK/NF-κB pathways and production of IL-12 and TNF-α by macrophages partially dependent on TLR9 but completely dependent on MyD88. In addition, B. abortus-derived CpG oligonucleotides required TLR9 to promote IL-12 and TNF-α production by macrophages. By confocal microscopy, we demonstrated that TLR9 redistributed and colocalized with lysosomal-associated membrane protein-1 upon Brucella infection. Thus, B. abortus induced TLR9 traffic, leading to cell signaling activation and IL-12 and TNF-α production. Although TLR9 recognized Brucella CpG motifs, our results suggest a new pathway of B. abortus DNA-activating macrophages independent of TLR9. © Society for Leukocyte Biology.

  4. Vaccination with Brucella abortus rough mutant RB51 protects BALB/c mice against virulent strains of Brucella abortus, Brucella melitensis, and Brucella ovis.

    PubMed Central

    Jiménez de Bagüés, M P; Elzer, P H; Jones, S M; Blasco, J M; Enright, F M; Schurig, G G; Winter, A J

    1994-01-01

    Vaccination of BALB/c mice with live Brucella abortus RB51, a stable rough mutant, produced protection against challenge with virulent strains of Brucella abortus, Brucella melitensis, and Brucella ovis. Passive-transfer experiments indicated that vaccinated mice were protected against B. abortus 2308 through cell-mediated immunity, against B. ovis PA through humoral immunity, and against B. melitensis 16M through both forms of immunity. Live bacteria were required for the induction of protective cell-mediated immunity; vaccination with whole killed cells of strain RB51 failed to protect mice against B. abortus 2308 despite development of good delayed-type hypersensitivity reactions. Protective antibodies against the heterologous species were generated in vaccinated mice primarily through anamnestic responses following challenge infections. Growth of the antigenically unrelated bacterium Listeria monocytogenes in the spleens of vaccinated mice indicated that nonspecific killing by residual activated macrophages contributed minimally to protection. These results encourage the continued investigation of strain RB51 as an alternative vaccine against heterologous Brucella species. However, its usefulness against B. ovis would be limited if, as suggested here, epitopes critical for protective cell-mediated immunity are not shared between B. abortus and B. ovis. Images PMID:7927779

  5. Intracellular trafficking of Brucella abortus in J774 macrophages.

    PubMed

    Arenas, G N; Staskevich, A S; Aballay, A; Mayorga, L S

    2000-07-01

    Brucella abortus is a facultative intracellular bacterium capable of surviving inside professional and nonprofessional phagocytes. The microorganism remains in membrane-bound compartments that in several cell types resemble modified endoplasmic reticulum structures. To monitor the intracellular transport of B. abortus in macrophages, the kinetics of fusion of phagosomes with preformed lysosomes labeled with colloidal gold particles was observed by electron microscopy. The results indicated that phagosomes containing live B. abortus were reluctant to fuse with lysosomes. Furthermore, newly endocytosed material was not incorporated into these phagosomes. These observations indicate that the bacteria strongly affect the normal maturation process of macrophage phagosomes. However, after overnight incubation, a significant percentage of the microorganisms were found in large phagosomes containing gold particles, resembling phagolysosomes. Most of the Brucella bacteria present in phagolysosomes were not morphologically altered, suggesting that they can also resist the harsh conditions prevalent in this compartment. About 50% colocalization of B. abortus with LysoSensor, a weak base that accumulates in acidic compartments, was observed, indicating that the B. abortus bacteria do not prevent phagosome acidification. In contrast to what has been described for HeLa cells, only a minor percentage of the microorganisms were found in compartments labeled with monodansylcadaverine, a marker for autophagosomes, and with DiOC6 (3,3'-dihexyloxacarbocyanine iodide), a marker for the endoplasmic reticulum. These results indicate that B. abortus bacteria alter phagosome maturation in macrophages. However, acidification does occur in these phagosomes, and some of them can eventually mature to phagolysosomes.

  6. Brucella abortus Inhibits Major Histocompatibility Complex Class II Expression and Antigen Processing through Interleukin-6 Secretion via Toll-Like Receptor 2

    PubMed Central

    Barrionuevo, Paula; Cassataro, Juliana; Delpino, M. Victoria; Zwerdling, Astrid; Pasquevich, Karina A.; Samartino, Clara García; Wallach, Jorge C.; Fossati, Carlos A.; Giambartolomei, Guillermo H.

    2008-01-01

    The strategies that allow Brucella abortus to survive inside macrophages for prolonged periods and to avoid the immunological surveillance of major histocompatibility complex class II (MHC-II)-restricted gamma interferon (IFN-γ)-producing CD4+ T lymphocytes are poorly understood. We report here that infection of THP-1 cells with B. abortus inhibited expression of MHC-II molecules and antigen (Ag) processing. Heat-killed B. abortus (HKBA) also induced both these phenomena, indicating the independence of bacterial viability and involvement of a structural component of the bacterium. Accordingly, outer membrane protein 19 (Omp19), a prototypical B. abortus lipoprotein, inhibited both MHC-II expression and Ag processing to the same extent as HKBA. Moreover, a synthetic lipohexapeptide that mimics the structure of the protein lipid moiety also inhibited MHC-II expression, indicating that any Brucella lipoprotein could down-modulate MHC-II expression and Ag processing. Inhibition of MHC-II expression and Ag processing by either HKBA or lipidated Omp19 (L-Omp19) depended on Toll-like receptor 2 and was mediated by interleukin-6. HKBA or L-Omp19 also inhibited MHC-II expression and Ag processing of human monocytes. In addition, exposure to the synthetic lipohexapeptide inhibited Ag-specific T-cell proliferation and IFN-γ production of peripheral blood mononuclear cells from Brucella-infected patients. Together, these results indicate that there is a mechanism by which B. abortus may prevent recognition by T cells to evade host immunity and establish a chronic infection. PMID:17984211

  7. [Preparation and characterization of follwing the national standard anti-Brucella abortus serum, bovine].

    PubMed

    Li, Cui; Guan, Fushi; Dai, Zhihong; Jiang, Hui; Wen, Fang; Lu, Lianshou; Wang, Zaishi

    2011-05-01

    To prepare anti-Brucella abortus serum used for calibrate the agglutination test follwing the national standard, 4 anti-Brucella abortus sera were obtained from 4 cows infected with Brucella abortus naturally. By potency testing, the third serum was selected. Sterility, vaccum degree, residual moisture, uniformity and stability of this standard material were tested and proved to meet the national standard. Referring to the international standard, RBT (Rose-Bengal plate agglutination test), SAT (standard tube agglutination) and CFT (complement fixation test) titers of this standard material were measured to be 1:160 "+" 1:2 400 "++" and 1:800 "++", which are identical with the collaborative assay results. International unit of the standard material is 4 000 IU/mL.

  8. Effectiveness of Brucella abortus lipopolysaccharide as an adjuvant for tuberculin PPD.

    PubMed

    Jamalan, Mostafa; Ardestani, Susan Kaboudanian; Zeinali, Majid; Mosaveri, Nader; Mohammad Taheri, Mohammad

    2011-01-01

    Bacterial lipopolysaccharide (LPS) has T-helper 1 (Th1) immunostimulatory activities but because of toxicity and pyrogenicity cannot be used as an adjuvant. Brucella abortus LPS has less toxicity and no pyrogenic properties in comparison to other bacterial LPS. In the current study, the immunostimulatory properties of B. abortus LPS were evaluated for its adjuvant activity. Tuberculin purified protein derivative (PPD) from Mycobacterium tuberculosis was extracted and after anion-exchange chromatography on Q-sepharose column, two fractions (17 and 23), which dominantly contained 30- and 70-kDa antigens, were collected for immunological studies. BALB/c mice were immunized with four different antigen preparations (BCG, PPD, 17th and 23rd PPD fractions) along with complete Freund's adjuvant or B. abortus LPS. The T-cell immune response of mice was assessed by measurement of Th1-type cytokine (IFN-γ) and Th2-type cytokines (IL-5 and IL-10) levels. Also, the humoral immunity was evaluated by measuring the specific IgG levels. Our results showed that immunization of mice with 17th PPD fraction along with B. abortus LPS can induce a Th1-type cytokine response characterized with a high IFN-γ/IL-5 ratio, while immunization with PPD or 23rd PPD fraction along with the same adjuvant resulted to a mixed Th1/Th2-type cytokine response. Copyright © 2010 The International Association for Biologicals. Published by Elsevier Ltd. All rights reserved.

  9. Occurrence of oxidative stress in dairy cows seropositives for Brucella abortus.

    PubMed

    Perin, Géssica; Fávero, Juscivete F; Severo, Diego R T; Silva, Anielen D; Machado, Gustavo; Araújo, Hugo L; Lilenbaum, Walter; Morsch, Vera M; Schetinger, Maria Rosa C; Jordão, Ricardo S; Stefani, Lenita M; Bottari, Nathieli B; Da Silva, Aleksandro S

    2017-09-01

    Bovine brucellosis is an important zoonotic disease caused by the bacterium Brucella abortus that leads to economic losses due to animal discard and commercial restrictions. Since positive animals for brucellosis are culled, little is known about the pathogenesis of this disease. Therefore, the aims of this study were to evaluate possible changes in the activity of deaminase adenosine (ADA) and the oxidative stress in cows seropositives for brucellosis (Experiment I), and to evaluate the seroprevalence of B. abortus in dairy cows from the Western state of Santa Catarina, Southern Brazil (Experiment II). The Experiment I evaluated 20 pregnant cows: ten seropositives for B. abortus and ten seronegatives that were used as controls. The ADA activity and markers of oxidative stress (TBARS, catalase (CAT) and superoxide dismutase (SOD)) were evaluated in these animals. A reduction in the activity of ADA and catalase enzymes in seropositive animals was observed (p < 0.001). Conversely, there was an increase in TBARS levels and superoxide dismutase activity in cows infected by B. abortus (p < 0.001). The presence of oxidative stress and a reduction of ADA might be related to the modulation of the inflammatory response. The experiment II was performed due to a high number of herds with restrictions imposed by cases of brucellosis in the state of Santa Catarina in the last two years, and thus, the seroprevalence for B. abortus was evaluated in 1242 serum samples of cows of 69 herds. The serodiagnosis was performed using two tests: buffered acidified antigen and 2-mercaptoethanol. However, none of the serum samples were positive for B. abortus. Although we did not find seropositive animals for brucellosis in our study, the disease still requires continued surveillance, due to its economic impact, and to the oxidative stress caused by it, which may have contributed to cases of abortion in three seropositive cows (Experiment I) in the final third of the gestation

  10. Can Chlamydia abortus be transmitted by embryo transfer in goats?

    PubMed

    Oseikria, M; Pellerin, J L; Rodolakis, A; Vorimore, F; Laroucau, K; Bruyas, J F; Roux, C; Michaud, S; Larrat, M; Fieni, F

    2016-10-01

    The objectives of this study were to determine (i) whether Chlamydia abortus would adhere to or penetrate the intact zona pellucida (ZP-intact) of early in vivo-derived caprine embryos, after in vitro infection; and (ii) the efficacy of the International Embryo Transfer Society (IETS) washing protocol for bovine embryos. Fifty-two ZP-intact embryos (8-16 cells), obtained from 14 donors were used in this experiment. The embryos were randomly divided into 12 batches. Nine batches (ZP-intact) of five embryos were incubated in a medium containing 4 × 10(7)Chlamydia/mL of AB7 strain. After incubation for 18 hours at 37 °C in an atmosphere of 5% CO2, the embryos were washed in batches in 10 successive baths of a phosphate buffer saline and 5% fetal calf serum solution in accordance with IETS guidelines. In parallel, three batches of ZP-intact embryos were used as controls by being subjected to similar procedures but without exposure to C. abortus. The 10 wash baths were collected separately and centrifuged for 1 hour at 13,000 × g. The washed embryos and the pellets of the 10 centrifuged wash baths were frozen at -20 °C before examination for evidence of C. abortus using polymerase chain reaction. C. abortus DNA was found in all of the infected batches of ZP-intact embryos (9/9) after 10 successive washes. It was also detected in the 10th wash fluid for seven batches of embryos, whereas for the two other batches, the last positive wash bath was the eighth and the ninth, respectively. In contrast, none of the embryos or their washing fluids in the control batches were DNA positive. These results report that C. abortus adheres to and/or penetrates the ZP of in vivo caprine embryos after in vitro infection, and that the standard washing protocol recommended by the IETS for bovine embryos, failed to remove it. The persistence of these bacteria after washing makes the embryo a potential means of transmission of the bacterium during embryo transfer from

  11. Identification of the 1B vaccine strain of Chlamydia abortus in aborted placentas during the investigation of toxaemic and systemic disease in sheep.

    PubMed

    Sargison, N D; Truyers, I G R; Howie, F E; Thomson, J R; Cox, A L; Livingstone, M; Longbottom, D

    2015-09-01

    One hundred and forty Cheviot and 100 Suffolk cross Mule primiparous 1-2-year-old ewes, from a flock of about 700 ewes, were vaccinated with an attenuated live 1B strain Chlamydia abortus vaccine about 4 weeks before ram introduction (September 2011). Between 08 March and 01 April 2012, 50 2-year-old ewes aborted and 29 of these died, despite antimicrobial and anti-inflammatory treatment and supportive care. Seven fetuses and three placentae from five 2-year-old ewes were submitted for pathological investigation. The aborted fetuses showed stages of autolysis ranging from being moderately fresh to putrefaction. Unusual, large multifocal regions of thickened membranes, with a dull red granular surface and moderate amounts of grey-white surface exudate were seen on each of the placentae. Intracellular, magenta-staining, acid fast inclusions were identified in Ziehl Neelsen-stained placental smears. Immunohistochemistry for Chlamydia-specific lipopolysaccharide showed extensive positive labelling of the placental epithelia. Molecular analyses of the aborted placentae demonstrated the presence of the 1B vaccine-type strain of C. abortus and absence of any wild-type field strain. The vaccine strain bacterial load of the placental tissue samples was consistent with there being an association between vaccination and abortion. Initial laboratory investigations resulted in a diagnosis of chlamydial abortion. Further investigations led to the identification of the 1B vaccine strain of C. abortus in material from all three of the submitted aborted placentae. Timely knowledge and understanding of any potential problems caused by vaccination against C. abortus are prerequisites for sustainable control of chlamydial abortion. This report describes the investigation of an atypical abortion storm in sheep, and describes the identification of the 1B vaccine strain of C. abortus in products of abortion. The significance of this novel putative association between the vaccine strain

  12. Improved immunogenicity of tetanus toxoid by Brucella abortus S19 LPS adjuvant.

    PubMed

    Mohammadi, Mohsen; Kianmehr, Zahra; Kaboudanian Ardestani, Sussan; Gharegozlou, Behnaz

    2014-09-01

    Adjuvants are used to increase the immunogenicity of new generation vaccines, especially those based on recombinant proteins. Despite immunostimulatory properties, the use of bacterial lipopolysaccharide (LPS) as an adjuvant has been hampered due to its toxicity and pyrogenicity. Brucella abortus LPS is less toxic and has no pyrogenic properties compared to LPS from other gram negative bacteria. To evaluate the adjuvant effect of B. abortus (vaccine strain, S19) LPS for tetanus toxoid antigen (TT) and to investigate the protective effect of different tetanus vaccine preparations. LPS was extracted and purified from B. abortus S19 and KDO, glycan, phosphate content, and protein contamination were measured. Adipic acid dihydrazide (ADH) was used as a linker for conjugation of TT to LPS. Different amounts of B. abortus LPS, TT, TT conjugated with LPS, and TT mixed with LPS or complete Freund's adjuvant (CFA) were injected into mice and antibody production against TT was measured. The protective effect of induced antibodies was determined by LD50. Immunization of mice with TT+LPS produced the highest anti-TT antibody titer in comparison to the group immunized with TT without any adjuvant or the groups immunized with TT-LPS or TT+CFA. Tetanus toxid-S19 LPS also produced a 100% protective effect against TT in immunized mice. These data indicate that B. abortus LPS enhances the immune responses to TT and suggest the possible use of B. abortus LPS as an adjuvant in vaccine preparations.

  13. In Vitro Antibacterial Effects of Five Volatile Oil Extracts Against Intramacrophage Brucella Abortus 544

    PubMed Central

    Al-Mariri, Ayman; Saour, George; Hamou, Razan

    2012-01-01

    Background: Brucella abortus is a gram-negative facultative intracellular bacterium that can cause a highly contagious disease in sheep, goats, cattle and one-humped camels. It is responsible for one of the most important zoonosis in human. The aim of this study was to evaluate the role of Mentha piperita, Origanum majorana, Citrus lemon, Cinnamomum verum and Myristica fragrans essential volatile oil extracts on human macrophages infected by B. abortus 544. Methods: Essential volatile oil extracts from M. piperita, O. majorana, C. lemon, C. verum and M. fragrans were extracted. Human macrophages were cultured at a density of 2×105 cells per well in sterile 96-well microtiter plates, and infected with B. abortus 544 at a ratio of 1:100 bacteria/cell. Then essential volatile oil extracts were added at a concentration of 1%. At specified times; cells were washed, lysed with 0.1% Triton, and plated on 2YT agar to determine the number of intracellular bacteria. Results: Cinnamomum verum volatile oil at a concentration of 1% had the highest antibacterial activity against B. abortus 544 inside human macrophages. Its inhibitory effect observed from 24 h and continued till 144 h after the infection. Moreover, C. verum (0.1%) in combination with 1% concentration of M. piperita, O. majorana, C. lemon or M. fragrans volatile oil extracts produced a synergistic inhibitory effect against B. abortus 544. Conclusion: The results indicate that, among the five selected oil extracts, C. verum volatile oil applied either separately or in combination with other oil extracts had the most effective antimicrobial activity against Brucella. PMID:23115441

  14. Modelling and mutational analysis of Aspergillus nidulans UreA, a member of the subfamily of urea/H+ transporters in fungi and plants

    PubMed Central

    Sanguinetti, Manuel; Amillis, Sotiris; Pantano, Sergio; Scazzocchio, Claudio; Ramón, Ana

    2014-01-01

    We present the first account of the structure–function relationships of a protein of the subfamily of urea/H+ membrane transporters of fungi and plants, using Aspergillus nidulans UreA as a study model. Based on the crystal structures of the Vibrio parahaemolyticus sodium/galactose symporter (vSGLT) and of the Nucleobase-Cation-Symport-1 benzylhydantoin transporter from Microbacterium liquefaciens (Mhp1), we constructed a three-dimensional model of UreA which, combined with site-directed and classical random mutagenesis, led to the identification of amino acids important for UreA function. Our approach allowed us to suggest roles for these residues in the binding, recognition and translocation of urea, and in the sorting of UreA to the membrane. Residues W82, Y106, A110, T133, N275, D286, Y388, Y437 and S446, located in transmembrane helixes 2, 3, 7 and 11, were found to be involved in the binding, recognition and/or translocation of urea and the sorting of UreA to the membrane. Y106, A110, T133 and Y437 seem to play a role in substrate selectivity, while S446 is necessary for proper sorting of UreA to the membrane. Other amino acids identified by random classical mutagenesis (G99, R141, A163, G168 and P639) may be important for the basic transporter's structure, its proper folding or its correct traffic to the membrane. PMID:24966243

  15. Differentiation of the serological response to Yersinia enterocolitica and Brucella abortus in cattle

    PubMed Central

    Corbel, M. J.; Cullen, G. A.

    1970-01-01

    The serological responses of cattle to inoculation with Brucella abortus and Yersinia enterocolitica type IX were compared. Complete cross-reactions were found in serum agglutination, antiglobulin, complement fixation and Rose Bengal plate tests. The cross-reaction between Br. abortus and Y. enterocolitica IX was confirmed by immunodiffusion tests. Although antibodies specific for each organism could also be detected by immunodiffusion tests with high titre rabbit or bovine sera, these tests were insufficiently sensitive for routine diagnostic use. A quantitative Rose Bengal plate test, using Rose Bengal stained Br. abortus and Y. enterocolitica IX, was developed which enabled the antibody responses to the two organisms to be differentiated. The specificity of this test was confirmed by cross-absorption experiments and its sensitivity was sufficient to permit evaluation of all bovine sera giving positive reactions to the serum agglutination test. ImagesFig. 1Fig. 2 PMID:4992575

  16. Deletion of nuoG from the Vaccine Candidate Mycobacterium bovis BCG ΔureC::hly Improves Protection against Tuberculosis

    PubMed Central

    Gengenbacher, Martin; Nieuwenhuizen, Natalie; Vogelzang, Alexis; Liu, Haipeng; Kaiser, Peggy; Schuerer, Stefanie; Lazar, Doris; Wagner, Ina; Mollenkopf, Hans-Joachim

    2016-01-01

    ABSTRACT The current tuberculosis (TB) vaccine, Mycobacterium bovis Bacillus Calmette-Guérin (BCG), provides insufficient protection against pulmonary TB. Previously, we generated a listeriolysin-expressing recombinant BCG strain, which to date has successfully completed phase I and phase IIa clinical trials. In an attempt to further improve efficacy, we deleted the antiapoptotic virulence gene nuoG, encoding NADH dehydrogenase 1 subunit G, from BCG ΔureC::hly. In vitro, deletion of nuoG unexpectedly led to strongly increased recruitment of the autophagosome marker LC3 to the engulfed vaccine, suggesting that nuoG also affects xenophagic pathways. In mice, BCG ΔureC::hly ΔnuoG vaccination was safer than BCG and improved protection over that of parental BCG ΔureC::hly, significantly reducing TB load in murine lungs, ameliorating pulmonary pathology, and enhancing immune responses. Transcriptome analysis of draining lymph nodes after vaccination with either BCG ΔureC::hly or BCG ΔureC::hly ΔnuoG demonstrated earlier and stronger induction of immune responses than that with BCG SSI and suggested upregulation of inflammasome activation and interferon-induced GTPases. In summary, BCG ΔureC::hly ΔnuoG is a promising next-generation TB vaccine candidate with excellent efficacy and safety. PMID:27222470

  17. Seroprevalence and molecular characterization of Chlamydia abortus in frozen fetal and placental tissues of aborting ewes in northeastern Algeria.

    PubMed

    Hireche, Sana; Ababneh, Mustafa Mohammed Kheir; Bouaziz, Omar; Boussena, Sabrina

    2016-02-01

    Enzootic abortion of ewes is one of the most serious health problems in sheep flocks worldwide. It has a significant economic impact because abortion, decrease in milk production and weak lambs. Besides, the bacteria is zoonotic. A cross-sectional study was conducted to determine the seroprevalence and risk factors associated with Chlamydia abortus infection in 552 ewes in Constantine using a C. abortus-specific indirect ELISA kit. Chlamydial DNA was investigated in ten ovine fetuses and eight placentas using PCR- restriction fragment length polymorphism (RFLP) and DNA sequencing. The study concluded that 7.2 % of ewes were seropositive and 33.3 % of sheep flocks had at least one seropositive ewe. Adjacent farmworker visits (OR = 7.667, 95 % CI (OR) = 2.307; 27.203) was defined as a risk factor. Deliveries of weak lambs (OR = 2.920, 95 % CI (OR) = 1.022; 8.342) and septicemia in lambs (OR = 9.971, 95 % CI (OR) = 2.383; 41.713) were significantly associated with chlamydial infection. PCR-RFLP analysis revealed positive signals to C. abortus in six fetuses and four placentas. Sequencing of the omp2 gene revealed that the Algerian strain is 96 % similar with C. abortus FAS strain. C. abortus plays a major role in abortion in northeastern Algeria. Appropriate control measures must be implemented to reduce economic losses and to avoid human contamination.

  18. Genetic stability of Brucella abortus isolates from an outbreak by multiple-locus variable-number tandem repeat analysis (MLVA16)

    PubMed Central

    2014-01-01

    Background Brucellosis caused by Brucella abortus is one of the most important zoonoses in the world. Multiple-locus variable-number tandem repeat analysis (MLVA16) has been shown be a useful tool to epidemiological traceback studies in B. abortus infection. Thus, the present study aimed (i) to evaluate the genetic diversity of B. abortus isolates from a brucellosis outbreak, and (ii) to investigate the in vivo stability of the MLVA16 markers. Results Three-hundred and seventy-five clinical samples, including 275 vaginal swabs and 100 milk samples, were cultured from a brucellosis outbreak in a cattle herd, which adopted RB51 vaccination and test-and-slaughter policies. Thirty-seven B. abortus isolates were obtained, eight from milk and twenty-nine from post-partum/abortion vaginal swabs, which were submitted to biotyping and genotyping by MLVA16. Twelve B. abortus isolates obtained from vaginal swabs were identified as RB51. Twenty four isolates, seven obtained from milk samples and seventeen from vaginal swabs, were identified as B. abortus biovar 3, while one isolate from vaginal swabs was identified as B. abortus biovar 1. Three distinct genotypes were observed during the brucellosis outbreak: RB observed in all isolates identified as RB51; W observed in all B. abortus biovar 3 isolates; and Z observed in the single B. abortus biovar 1 isolate. Epidemiological and molecular data show that the B. abortus biovar 1 genotype Z strain is not related to the B. abortus biovar 3 genotype W isolates, and represents a new introduction B. abortus during the outbreak. Conclusions The results of the present study on typing of multiple clinical B. abortus isolates from the same outbreak over a sixteen month period indicate the in vivo stability of MLVA16 markers, a low genetic diversity among B. abortus isolates and the usefulness of MLVA16 for epidemiological studies of bovine brucellosis. PMID:25015840

  19. Mutational analysis of the major soybean UreF paralogue involved in urease activation

    USDA-ARS?s Scientific Manuscript database

    In soybean, mutation at Eu2 or Eu3 eliminates the urease activities of both the embryo-specific and the tissue-ubiquitous (assimilatory) isozymes, encoded by Eu1 and Eu4, respectively. Eu3 encodes UreG, a GTP’ase necessary for proper emplacement of Ni and carbon dioxide in the urease active site. ...

  20. The Tuberculosis Vaccine Candidate Bacillus Calmette-Guérin ΔureC::hly Coexpressing Human Interleukin-7 or -18 Enhances Antigen-Specific T Cell Responses in Mice

    PubMed Central

    Rao, Martin; Vogelzang, Alexis; Kaiser, Peggy; Schuerer, Stefanie; Kaufmann, Stefan H. E.; Gengenbacher, Martin

    2013-01-01

    Bacillus Calmette–Guérin (BCG), the only approved tuberculosis vaccine, provides only limited protection. Previously, we generated a recombinant derivative (BCG ΔureC::hly), which secretes the pore-forming toxin listeriolysin O (LLO) of Listeria monocytogenes. This vaccine shows superior protection against tuberculosis in preclinical models and is safe in humans. Here we describe two new vaccine strains which express human interleukin-7 (hIL)-7 or hIL-18 in the genetic background of BCG ΔureC::hly to modulate specific T cell immunity. Both strains exhibited an uncompromised in vitro growth pattern, while inducing a proinflammatory cytokine profile in human dendritic cells (DCs). Human DCs harbouring either strain efficiently promoted secretion of IL-2 by autologous T cells in a coculture system, suggesting superior immunogenicity. BALB/c mice vaccinated with BCG ΔureC::hly, BCG ΔureC::hly_hIL7 or BCG ΔureC::hly_hIL18 developed a more robust Th1 response than after vaccination with parental BCG. Both strains provided significantly better protection than BCG in a murine Mycobacterium tuberculosis challenge model but efficacy remained comparable to that afforded by BCG ΔureC::hly. We conclude that expression of hIL-7 or hIL-18 enhanced specific T cell responses but failed to improve protection over BCG ΔureC::hly in mice. PMID:24236077

  1. European Chlamydia abortus livestock isolate genomes reveal unusual stability and limited diversity, reflected in geographical signatures.

    PubMed

    Seth-Smith, H M B; Busó, Leonor Sánchez; Livingstone, M; Sait, M; Harris, S R; Aitchison, K D; Vretou, Evangelia; Siarkou, V I; Laroucau, K; Sachse, K; Longbottom, D; Thomson, N R

    2017-05-04

    Chlamydia abortus (formerly Chlamydophila abortus) is an economically important livestock pathogen, causing ovine enzootic abortion (OEA), and can also cause zoonotic infections in humans affecting pregnancy outcome. Large-scale genomic studies on other chlamydial species are giving insights into the biology of these organisms but have not yet been performed on C. abortus. Our aim was to investigate a broad collection of European isolates of C. abortus, using next generation sequencing methods, looking at diversity, geographic distribution and genome dynamics. Whole genome sequencing was performed on our collection of 57 C. abortus isolates originating primarily from the UK, Germany, France and Greece, but also from Tunisia, Namibia and the USA. Phylogenetic analysis of a total of 64 genomes shows a deep structural division within the C. abortus species with a major clade displaying limited diversity, in addition to a branch carrying two more distantly related Greek isolates, LLG and POS. Within the major clade, seven further phylogenetic groups can be identified, demonstrating geographical associations. The number of variable nucleotide positions across the sampled isolates is significantly lower than those published for C. trachomatis and C. psittaci. No recombination was identified within C. abortus, and no plasmid was found. Analysis of pseudogenes showed lineage specific loss of some functions, notably with several Pmp and TMH/Inc proteins predicted to be inactivated in many of the isolates studied. The diversity within C. abortus appears to be much lower compared to other species within the genus. There are strong geographical signatures within the phylogeny, indicating clonal expansion within areas of limited livestock transport. No recombination has been identified within this species, showing that different species of Chlamydia may demonstrate different evolutionary dynamics, and that the genome of C. abortus is highly stable.

  2. Typing Discrepancy Between Phenotypic and Molecular Characterization Revealing an Emerging Biovar 9 Variant of Smooth Phage-Resistant B. abortus Strain 8416 in China.

    PubMed

    Kang, Yao-Xia; Li, Xu-Ming; Piao, Dong-Ri; Tian, Guo-Zhong; Jiang, Hai; Jia, En-Hou; Lin, Liang; Cui, Bu-Yun; Chang, Yung-Fu; Guo, Xiao-Kui; Zhu, Yong-Zhang

    2015-01-01

    A newly isolated smooth colony morphology phage-resistant strain 8416 isolated from a 45-year-old cattle farm cleaner with clinical features of brucellosis in China was reported. The most unusual phenotype was its resistance to two Brucella phages Tbilisi and Weybridge, but sensitive to Berkeley 2, a pattern similar to that of Brucella melitensis biovar 1. VITEK 2 biochemical identification system found that both strain 8416 and B. melitensis strains shared positive ILATk, but negative in other B. abortus strains. However, routine biochemical and phenotypic characteristics of strain 8416 were most similar to that of B. abortus biovar 9 except CO2 requirement. In addition, multiple PCR molecular typing assays including AMOS-PCR, B. abortus special PCR (B-ab PCR) and a novel sub-biovar typing PCR, indicated that strain 8416 may belong to either biovar 3b or 9 of B. abortus. Surprisingly, further MLVA typing results showed that strain 8416 was most closely related to B. abortus biovar 3 in the Brucella MLVA database, primarily differing in 4 out of 16 screened loci. Therefore, due to the unusual discrepancy between phenotypic (biochemical reactions and particular phage lysis profile) and molecular typing characteristics, strain 8416 could not be exactly classified to any of the existing B. abortus biovars and might be a new variant of B. abortus biovar 9. The present study also indicates that the present phage typing scheme for Brucella sp. is subject to variation and the routine Brucella biovar typing needs further studies.

  3. Structure of Rv1848 (UreA), the Mycobacterium tuberculosis urease γ subunit

    PubMed Central

    Habel, Jeff E.; Bursey, Evan H.; Rho, Beom-Seop; Kim, Chang-Yub; Segelke, Brent W.; Rupp, Bernhard; Park, Min S.; Terwilliger, Thomas C.; Hung, Li-Wei

    2010-01-01

    The crystal structure of the urease γ subunit (UreA) from Mycobacterium tuberculosis, Rv1848, has been determined at 1.8 Å resolution. The asymmetric unit contains three copies of Rv1848 arranged into a homotrimer that is similar to the UreA trimer in the structure of urease from Klebsiella aerogenes. Small-angle X-ray scattering experiments indicate that the Rv1848 protein also forms trimers in solution. The observed homotrimer and the organization of urease genes within the M. tuberculosis genome suggest that M. tuberculosis urease has the (αβγ)3 composition observed for other bacterial ureases. The γ subunit may be of primary importance for the formation of the urease quaternary structure. PMID:20606272

  4. Identification of two novel immunodominant UreB CD4(+) T cell epitopes in Helicobacter pylori infected subjects.

    PubMed

    Yang, Wu-Chen; Chen, Li; Li, Hai-Bo; Li, Bin; Hu, Jian; Zhang, Jin-Yong; Yang, Shi-Ming; Zou, Quan-Ming; Guo, Hong; Wu, Chao

    2013-02-06

    An epitope-based vaccine is a promising option for treating Helicobacter pylori (H. pylori) infection. Epitope mapping is the first step in designing an epitope-based vaccine. A pivotal role of CD4(+) T cells in protection against H. pylori has been accepted, but few Th epitopes have been identified. In this study, two novel UreB CD4(+) T cell epitopes were identified using PBMCs obtained from two H. pylori infected subjects. We determined the restriction molecules by antibody blocking and used various Epstein-Barr virus-transformed B lymphocyte cell lines (BLCLs) with different HLA alleles as APCs to present peptides to CD4(+) T cells. These epitopes were DRB1*1404-restricted UreB(373-385) and DRB1*0803-restricted UreB(438-452). The T cells specific to these epitopes not only recognized autologous DCs loaded with recombinant UreB but also those pulsed with H. pylori whole cell lysates, suggesting that these epitope peptides are naturally processed. These epitopes have important value for designing an effective H. pylori vaccine. Copyright © 2012 Elsevier Ltd. All rights reserved.

  5. The bovine immune response to Brucella abortus I. A water soluble antigen precipitated by sera of some naturally infected cattle.

    PubMed Central

    Stemshorn, B; Nielsen, K

    1977-01-01

    Selected sera from cattle naturally infected with Brucella abortus precipitate water soluble antigens extracted by sonication from B. abortus. One of these antigens resembles antigen E (Baughn and Freeman) as it is excluded from Sephadex G-200 gels, migrates anodally when electrophoresed at pH 8.6, resists heating at 100 degrees C for ten minutes and appears to be susceptible to papain digestion. Precipitins specific for this antigen remained in sera from which all detectable Brucella agglutinating antibody had been removed by adsorption with live or heat killed B. abortus. The antigen has been extracted from smooth and rough strains of B abortus. Precipitins specific for this antigen have been detected in antisera produced against Brucella canis. Images Fig. 1. Fig. 2. Fig. 3. Fig. 4. PMID:405088

  6. Diversification and Distribution of Ruminant Chlamydia abortus Clones Assessed by MLST and MLVA.

    PubMed

    Siarkou, Victoria I; Vorimore, Fabien; Vicari, Nadia; Magnino, Simone; Rodolakis, Annie; Pannekoek, Yvonne; Sachse, Konrad; Longbottom, David; Laroucau, Karine

    2015-01-01

    Chlamydia abortus, an obligate intracellular bacterium, is the most common infectious cause of abortion in small ruminants worldwide and has zoonotic potential. We applied multilocus sequence typing (MLST) together with multiple-locus variable-number tandem repeat analysis (MLVA) to genotype 94 ruminant C. abortus strains, field isolates and samples collected from 1950 to 2011 in diverse geographic locations, with the aim of delineating C. abortus lineages and clones. MLST revealed the previously identified sequence types (STs) ST19, ST25, ST29 and ST30, plus ST86, a recently-assigned type on the Chlamydiales MLST website and ST87, a novel type harbouring the hemN_21 allele, whereas MLVA recognized seven types (MT1 to MT7). Minimum-spanning-tree analysis suggested that all STs but one (ST30) belonged to a single clonal complex, possibly reflecting the short evolutionary timescale over which the predicted ancestor (ST19) has diversified into three single-locus variants (ST86, ST87 and ST29) and further, through ST86 diversification, into one double-locus variant (ST25). ST descendants have probably arisen through a point mutation evolution mode. Interestingly, MLVA showed that in the ST19 population there was a greater genetic diversity than in other STs, most of which exhibited the same MT over time and geographical distribution. However, the evolutionary pathways of C. abortus STs seem to be diverse across geographic distances with individual STs restricted to particular geographic locations. The ST30 singleton clone displaying geographic specificity and represented by the Greek strains LLG and POS was effectively distinguished from the clonal complex lineage, supporting the notion that possibly two separate host adaptations and hence independent bottlenecks of C. abortus have occurred through time. The combination of MLST and MLVA assays provides an additional level of C. abortus discrimination and may prove useful for the investigation and surveillance of

  7. Pheno- and genotyping of Brucella abortus biovar 5 isolated from a water buffalo (Bubalus bubalis) fetus: First case reported in the Americas.

    PubMed

    Martínez, Diana; Thompson, Carolina; Draghi, Graciela; Canavesio, Vilma; Jacobo, Roberto; Zimmer, Patricia; Elena, Sebastián; Nicola, Ana M; de Echaide, Susana Torioni

    2014-09-17

    An isolate of Brucella spp. from an aborted water buffalo (Bubalus bubalis) fetus was characterized based on its pheno- and genotype. The phenotype was defined by carbon dioxide requirement, hydrogen sulfide production, sensitivity to thionin and basic fuchsin and agglutination with Brucella A and M monospecific antisera. The genotype was based on the amplification of the following genes: bcsp31, omp2ab, and eri and the species-specific localization of the insertion sequence IS711 in the Brucella chromosome via B. abortus-B. melitensis-B. ovis-B. suis (AMOS)-PCR. Unexpectedly, the isolate showed a phenotype different from B. abortus bv 1, the most prevalent strain in cattle in Argentina, and from vaccine strain 19, currently used in bovines and water buffaloes. Genotyping supported the phenotypic results, as the analysis of the omp2ab gene sequence showed an identical pattern to either B. abortus bv 5 or B. melitensis. Finally, the AMOS PCR generated a 1700-bp fragment from the isolate, different than those amplified from B. abortus bv 1 (498bp) and B. melitensis (731bp), confirming the presence of B. abortus bv 5. The OIE/FAO Reference Laboratory for Brucellosis confirmed this typing. This is the first report of B. abortus bv 5 from a water buffalo in the Americas. Copyright © 2014 Elsevier B.V. All rights reserved.

  8. Improving the molecular diagnosis of Chlamydia psittaci and Chlamydia abortus infection with a species-specific duplex real-time PCR.

    PubMed

    Opota, Onya; Jaton, Katia; Branley, James; Vanrompay, Daisy; Erard, Veronique; Borel, Nicole; Longbottom, David; Greub, Gilbert

    2015-10-01

    Chlamydia psittaci and Chlamydia abortus are closely related intracellular bacteria exhibiting different tissue tropism that may cause severe but distinct infection in humans. C. psittaci causes psittacosis, a respiratory zoonotic infection transmitted by birds. C. abortus is an abortigenic agent in small ruminants, which can also colonize the human placenta and lead to foetal death and miscarriage. Infections caused by C. psittaci and C. abortus are underestimated mainly due to diagnosis difficulties resulting from their strict intracellular growth. We developed a duplex real-time PCR to detect and distinguish these two bacteria in clinical samples. The first PCR (PCR1) targeted a sequence of the 16S-23S rRNA operon allowing the detection of both C. psittaci and C. abortus. The second PCR (PCR2) targeted the coding DNA sequence CPSIT_0607 unique to C. psittaci. The two PCRs showed 100 % detection for ≥ 10 DNA copies per reaction (1000 copies ml(- 1)). Using a set of 120 samples, including bacterial reference strains, clinical specimens and infected cell culture material, we monitored 100 % sensitivity and 100 % specificity for the detection of C. psittaci and C. abortus for PCR1. When PCR1 was positive, PCR2 could discriminate C. psittaci from C. abortus with a positive predictive value of 100 % and a negative predictive value of 88 %. In conclusion, this new duplex PCR represents a low-cost and time-saving method with high-throughput potential, expected to improve the routine diagnosis of psittacosis and pregnancy complication in large-scale screening programs and also during outbreaks.

  9. In vitro antibacterial effects of five volatile oil extracts against intramacrophage Brucella abortus 544.

    PubMed

    Al-Mariri, Ayman; Saour, George; Hamou, Razan

    2012-06-01

    Brucellaabortus is a gram-negative facultative intracellular bacterium that can cause a highly contagious disease in sheep, goats, cattle and one-humped camels. It is responsible for one of the most important zoonosis in human. The aim of this study was to evaluate the role of Mentha piperita, Origanum majorana, Citrus lemon, Cinnamomum verum and Myristica fragrans essential volatile oil extracts on human macrophages infected by B. abortus 544. Essential volatile oil extracts from M. piperita, O. majorana, C. lemon, C. verum and M. fragrans were extracted. Human macrophages were cultured at a density of 2×10(5) cells per well in sterile 96-well microtiter plates, and infected with B. abortus 544 at a ratio of 1:100 bacteria/cell. Then essential volatile oil extracts were added at a concentration of 1%. At specified times; cells were washed, lysed with 0.1% Triton, and plated on 2YT agar to determine the number of intracellular bacteria. Cinnamomum verum volatile oil at a concentration of 1% had the highest antibacterial activity against B. abortus 544 inside human macrophages. Its inhibitory effect observed from 24 h and continued till 144 h after the infection. Moreover, C. verum (0.1%) in combination with 1% concentration of M. piperita, O. majorana, C. lemon or M. fragrans volatile oil extracts produced a synergistic inhibitory effect against B. abortus 544. The results indicate that, among the five selected oil extracts, C. verum volatile oil applied either separately or in combination with other oil extracts had the most effective antimicrobial activity against Brucella.

  10. GLUTAMATE METABOLISM IN BRUCELLA ABORTUS STRAINS OF LOW AND HIGH VIRULENCE

    PubMed Central

    Dasinger, B. L.; Wilson, J. B.

    1962-01-01

    Dasinger, B. L. (University of Wisconsin, Madison) and J. B. Wilson. Glutamate metabolism in Brucella abortus strains of low and high virulence. J. Bacteriol. 84:911–915. 1962.—Brucella abortus strains of low virulence oxidize glutamate at a high rate, whereas strains of high virulence oxidize glutamate at a relatively low rate. Results indicated that this observation was not related to differences in pathway of glutamate oxidation or to differences in total enzyme activity. Permeability studies showed that the maximal rates of glutamate accumulation were 2.8 μmoles per 2 min per g (wet wt) for a strain of high virulence and 6.2 μmoles per 2 min per g for a strain of low virulence, but equal intracellular steady-state concentrations were attained by both types of strains. Evidence is presented which suggests that the site of glutamate oxidation is separate from the pool of glutamate being measured. Unequal rates of permeability at these sites could be the reason for the differences in rate of glutamate oxidation. PMID:14028057

  11. The EdUReP approach plus manual therapy for the management of insertional Achilles tendinopathy.

    PubMed

    Sartorio, Francesco; Zanetta, Anna; Ferriero, Giorgio; Bravini, Elisabetta; Vercelli, Stefano

    2018-05-01

    Insertional Achilles tendinopathy (IAT) is a challenging overuse disorder. The aim of this case report was to study the feasibility of a comprehensive rehabilitative approach according to the Education, Unloading, Reloading, and Prevention (EdUReP) framework combined with Instrument-Augmented Soft Tissue Mobilization (I-ASTM). An active 51-year-old man patient with chronic IAT was studied. Clinical assessment battery was composed by visual analogue scale for pain during the Achilles tendon palpation test, passive straight leg raise test, single leg hop test, Patient-Specific Functional Scale, and Foot and Ankle Ability Measure. The patient was treated over a 8 weeks period using the EdUReP guidelines plus 8 sessions of I-ASTM, applied with a solid instrument to the Achilles tendon and to the muscle fibrotic areas previously identified during evaluation. Clinically significant improvements were observed in all outcome measures, and a resume of patient's usual sports activities without pain or limitations was possible after treatment. Results lasted over a 6-month follow-up. To the best of our knowledge, this is the first study applying a comprehensive approach based on accurate physical assessment, and using the EdUReP theoretical model. The combination of the EdUReP model and manual therapy was effective in resolving the patient's symptoms and restore his usual sport activities. While these results cannot be generalized, the present findings could provide a valuable foundation for future researches.

  12. Expression and display of UreA of Helicobacter acinonychis on the surface of Bacillus subtilis spores.

    PubMed

    Hinc, Krzysztof; Isticato, Rachele; Dembek, Marcin; Karczewska, Joanna; Iwanicki, Adam; Peszyńska-Sularz, Grazyna; De Felice, Maurilio; Obuchowski, Michał; Ricca, Ezio

    2010-01-18

    The bacterial endospore (spore) has recently been proposed as a new surface display system. Antigens and enzymes have been successfully exposed on the surface layers of the Bacillus subtilis spore, but only in a few cases the efficiency of expression and the effective surface display and have been determined. We used this heterologous expression system to produce the A subunit of the urease of the animal pathogen Helicobater acinonychis. Ureases are multi-subunit enzymes with a central role in the virulence of various bacterial pathogens and necessary for colonization of the gastric mucosa by the human pathogen H. pylori. The urease subunit UreA has been recognized as a major antigen, able to induce high levels of protection against challenge infections. We expressed UreA from H. acinonychis on the B. subtilis spore coat by using three different spore coat proteins as carriers and compared the efficiency of surface expression and surface display obtained with the three carriers. A combination of western-, dot-blot and immunofluorescence microscopy allowed us to conclude that, when fused to CotB, UreA is displayed on the spore surface (ca. 1 x 10(3) recombinant molecules per spore), whereas when fused to CotC, although most efficiently expressed (7-15 x 10(3) recombinant molecules per spore) and located in the coat layer, it is not displayed on the surface. Experiments with CotG gave results similar to those with CotC, but the CotG-UreA recombinant protein appeared to be partially processed. UreA was efficiently expressed on the spore coat of B. subtilis when fused to CotB, CotC or CotG. Of these three coat proteins CotC allows the highest efficiency of expression, whereas CotB is the most appropriate for the display of heterologous proteins on the spore surface.

  13. Brucellosis vaccines based on the open reading frames from genomic island 3 of Brucella abortus.

    PubMed

    Gómez, Leonardo; Alvarez, Francisco; Betancur, Daniel; Oñate, Angel

    2018-05-17

    Brucella abortus is the etiological agent of brucellosis, a zoonotic disease affecting cattle and humans. This disease has been partially controlled in cattle by immunization with live attenuated B. abortus S19 and RB51 strains. However, use of these vaccine strains has been associated with safety issues in animals and humans. New vaccines have since emerged in the prevention of brucellosis, particularly DNA vaccines, which have shown effectiveness and a good safety profile. Their protection efficacy in mice is associated with the induction of Th1 type and cytotoxic T cell mediated immune response against structural antigens and virulence factors expressed during B. abortus infection. Some antigenic candidate for vaccine design against brucellosis (mainly DNA vaccines) have been obtained from genomic island 3 (GI-3) of B. abortus, which encodes several open reading frames (ORFs) involved in the intracellular survival and virulence of this pathogen. The immunogenicity and protection conferred by these DNA vaccines in a murine model is reviewed in this article, suggesting that some of them could be safe and effective vaccine candidates against to prevent B. abortus infection. Copyright © 2018 Elsevier Ltd. All rights reserved.

  14. Brucella abortus is Prevalent in Both Humans and Animals in Bangladesh.

    PubMed

    Rahman, A K M A; Saegerman, C; Berkvens, D; Melzer, F; Neubauer, H; Fretin, D; Abatih, E; Dhand, N; Ward, M P

    2017-08-01

    To determine the role of different Brucella (B.) spp. in Bangladesh, 62 animal samples and 500 human sera were tested. Animal samples from cattle, goats and sheep (including milk, bull semen, vaginal swabs and placentas) were cultured for Brucella spp. Three test-positive human sera and all animal samples were screened by Brucella genus-specific real-time PCR (RT-PCR), and positive samples were then tested by IS711 RT-PCR to detect B. abortus and B. melitensis DNA. Only B. abortus DNA was amplified from 13 human and six animal samples. This is the first report describing B. abortus as the aetiological agent of brucellosis in occupationally exposed humans in Bangladesh. Of note is failure to detect B. melitensis DNA, the species most often associated with human brucellosis worldwide. Further studies are required to explore the occurrence of Brucella melitensis in Bangladesh. © 2017 Blackwell Verlag GmbH.

  15. Effects of gamma radiation and azathioprine on Brucella abortus infection in BALB/c mice

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Elzer, P.H.; Rowe, G.E.; Enright, F.M.

    Sublethal irradiation of BALB/c mice 4 hours prior to inoculation with 5 {times} 10(4) virulent Brucella abortus, caused significant (P less than 0.01) reductions in bacterial numbers in comparison with numbers in unirradiated controls. Numbers of brucellae in the spleen were significantly lower by 5 days after inoculation and decreased thereafter, so that at 2 and 3 weeks after inoculation, there were up to 1,000-fold fewer organisms in the spleen of irradiated mice. The number of brucellae in the spleen increased in irradiated mice thereafter. The course of events in the liver was similar, but developed more slowly, and peakmore » differences in bacterial numbers were about 1 log less. These phenomena were not attributable to differences in implantation of brucellae in the liver or spleen, nor to an abnormal distribution of organisms in other organs of irradiated mice. Irradiation of mice during the plateau phase of infection also resulted in significant (P less than 0.05) reductions in bacterial counts in the spleen during the succeeding 4 weeks. Macrophage activation in the spleen, measured by a Listeria monocytogenes-killing assay, was significantly (P less than 0.01) increased by irradiation alone at 1 week after inoculation and at that time was significantly (P less than 0.01) greater in B abortus-infected, irradiated mice than in B abortus-infected controls. Histologic, cytologic, and immunologic studies revealed that the decrease in numbers of organisms between 1 and 2 weeks after inoculation in irradiated mice occurred at a time when their immune response to B abortus was suppressed and when numbers of neutrophils and monocytes infiltrating the spleen were significantly (P less than 0.01) diminished.« less

  16. Comparative analysis of the early transcriptome of Brucella abortus - infected monocyte-derived macrophages from cattle naturally resistant or susceptible to brucellosis

    PubMed Central

    Rossetti, C.A.; Galindo, C.L.; Everts, R.E.; Lewin, H.A.; Garner, H.R.; Adams, L.G.

    2010-01-01

    Brucellosis is a worldwide zoonotic infectious disease that has a significant economic impact on animal production and human public health. We characterized the gene expression profile of B. abortus-infected monocyte-derived macrophages (MDMs) from naïve cattle naturally resistant (R) or susceptible (S) to brucellosis using a cDNA microarray technology. Our data indicate that 1) B. abortus induced a slightly increased genome activation in R MDMs and a down-regulated transcriptome in S MDMs, during the onset of infection, 2) R MDMs had the ability to mount a type 1 immune response against B. abortus infection which was impaired in S cells, and 3) the host cell activity was not altered after 12h post-B. abortus infection in R MDMs while the cell cycle was largely arrested in infected S MDMs at 12h p.i. These results contribute to understand of how host responses may be manipulated to prevent infection by brucellae. PMID:20932540

  17. Gene Discovery through Genomic Sequencing of Brucella abortus

    PubMed Central

    Sánchez, Daniel O.; Zandomeni, Ruben O.; Cravero, Silvio; Verdún, Ramiro E.; Pierrou, Ester; Faccio, Paula; Diaz, Gabriela; Lanzavecchia, Silvia; Agüero, Fernán; Frasch, Alberto C. C.; Andersson, Siv G. E.; Rossetti, Osvaldo L.; Grau, Oscar; Ugalde, Rodolfo A.

    2001-01-01

    Brucella abortus is the etiological agent of brucellosis, a disease that affects bovines and human. We generated DNA random sequences from the genome of B. abortus strain 2308 in order to characterize molecular targets that might be useful for developing immunological or chemotherapeutic strategies against this pathogen. The partial sequencing of 1,899 clones allowed the identification of 1,199 genomic sequence surveys (GSSs) with high homology (BLAST expect value < 10−5) to sequences deposited in the GenBank databases. Among them, 925 represent putative novel genes for the Brucella genus. Out of 925 nonredundant GSSs, 470 were classified in 15 categories based on cellular function. Seven hundred GSSs showed no significant database matches and remain available for further studies in order to identify their function. A high number of GSSs with homology to Agrobacterium tumefaciens and Rhizobium meliloti proteins were observed, thus confirming their close phylogenetic relationship. Among them, several GSSs showed high similarity with genes related to nodule nitrogen fixation, synthesis of nod factors, nodulation protein symbiotic plasmid, and nodule bacteroid differentiation. We have also identified several B. abortus homologs of virulence and pathogenesis genes from other pathogens, including a homolog to both the Shda gene from Salmonella enterica serovar Typhimurium and the AidA-1 gene from Escherichia coli. Other GSSs displayed significant homologies to genes encoding components of the type III and type IV secretion machineries, suggesting that Brucella might also have an active type III secretion machinery. PMID:11159979

  18. A B lymphocyte mitogen is a Brucella abortus virulence factor required for persistent infection

    PubMed Central

    Spera, Juan Manuel; Ugalde, Juan Esteban; Mucci, Juan; Comerci, Diego J.; Ugalde, Rodolfo Augusto

    2006-01-01

    Microbial pathogens with the ability to establish chronic infections have evolved strategies to actively modulate the host immune response. Brucellosis is a disease caused by a Gram-negative intracellular pathogen that if not treated during the initial phase of the infection becomes chronic as the bacteria persist for the lifespan of the host. How this pathogen and others achieve this action is a largely unanswered question. We report here the identification of a Brucella abortus gene (prpA) directly involved in the immune modulation of the host. PrpA belongs to the proline-racemase family and elicits a B lymphocyte polyclonal activation that depends on the integrity of its proline-racemase catalytic site. Stimulation of splenocytes with PrpA also results in IL-10 secretion. Construction of a B. abortus-prpA mutant allowed us to assess the contribution of PrpA to the infection process. Mice infected with B. abortus induced an early and transient nonresponsive status of splenocytes to both Escherichia coli LPS and ConA. This phenomenon was not observed when mice were infected with a B. abortus-prpA mutant. Moreover, the B. abortus-prpA mutant had a reduced capacity to establish a chronic infection in mice. We propose that an early and transient nonresponsive immune condition of the host mediated by this B cell polyclonal activator is required for establishing a successful chronic infection by Brucella. PMID:17053080

  19. Comparison of Buffered, Acidified Plate Antigen to Standard Serologic Tests for the Detection of Serum Antibodies to Brucella abortus in Elk (Cervus canadensis).

    PubMed

    Clarke, P Ryan; Edwards, William H; Hennager, Steven G; Block, Jean F; Yates, Angela M; Ebel, Eric; Knopp, Douglas J; Fuentes-Sanchez, Antonio; Jennings-Gaines, Jessica; Kientz, Rebecca L; Simunich, Marilyn

    2015-07-01

    Brucellosis (caused by the bacterium Brucella abortus) is a zoonotic disease endemic in wild elk (Cervus canadensis) of the Greater Yellowstone Ecosystem, US. Because livestock and humans working with elk or livestock are at risk, validated tests to detect the B. abortus antibody in elk are needed. Using the κ-statistic, we evaluated the buffered, acidified plate antigen (BAPA) assay for agreement with the results of the four serologic tests (card test [card], complement fixation test [CF], rivanol precipitation plate agglutination test [RIV], standard plate agglutination test [SPT]) that are approved by the US Department of Agriculture for the detection of the B. abortus antibody in elk. From 2006 to 2010, serum samples collected from elk within B. abortus-endemic areas (n = 604) and nonendemic areas (n = 707) and from elk culture-positive for B. abortus (n = 36) were split and blind tested by four elk serum diagnostic laboratories. κ-Values showed a high degree of agreement for the card (0.876), RIV (0.84), and CF (0.774) test pairings and moderate agreement for the SPT (0.578). Sensitivities for the BAPA, card, RIV, CF, and SPT were 0.859, 0.839, 0.899, 1.00, and 0.813, whereas specificities were 0.986, 0.993, 0.986, 0.98, and 0.968, respectively. The positive predictive values and the negative predictive values were calculated for 2.6%, 8.8%, and 16.2% prevalence levels. These findings suggest the BAPA test is a suitable screening test for the B. abortus antibodies in elk.

  20. Proteomic analysis of Brucella abortus cell envelope and identification of immunogenic candidate proteins for vaccine development.

    PubMed

    Connolly, Joseph P; Comerci, Diego; Alefantis, Timothy G; Walz, Alexander; Quan, Marian; Chafin, Ryan; Grewal, Paul; Mujer, Cesar V; Ugalde, Rodolfo A; DelVecchio, Vito G

    2006-07-01

    Brucella abortus is the etiologic agent of bovine brucellosis and causes a chronic disease in humans known as undulant fever. In livestock the disease is characterized by abortion and sterility. Live, attenuated vaccines such as S19 and RB51 have been used to control the spread of the disease in animals; however, they are considered unsafe for human use and they induce abortion in pregnant cattle. For the development of a safer and equally efficacious vaccine, immunoproteomics was utilized to identify novel candidate proteins from B. abortus cell envelope (CE). A total of 163 proteins were identified using 2-DE with MALDI-TOF MS and LC-MS/MS. Some of the major protein components include outer-membrane protein (OMP) 25, OMP31, Omp2b porin, and 60 kDa chaperonin GroEL. 2-DE Western blot analyses probed with antiserum from bovine and a human patient infected with Brucella identified several new immunogenic proteins such as fumarate reductase flavoprotein subunit, F0F1-type ATP synthase alpha subunit, and cysteine synthase A. The elucidation of the immunome of B. abortus CE identified a number of candidate proteins for developing vaccines against Brucella infection in bovine and humans.

  1. Protection of BALB/C mice against Brucella abortus 544 challenge by vaccination with combination of recombinant human serum albumin-l7/l12 (Brucella abortus ribosomal protein) and lipopolysaccharide.

    PubMed

    Pakzad, Iraj; Rezaee, Abbas; Rasaee, Mohammad J; Hossieni, Ahmad Zavaran; Tabbaraee, Bahman; Kazemnejad, Anoshirvan

    2010-01-01

    The immunogenic Brucella abortus ribosomal protein L7/L12 and Lipopolysaccharide (LPS) are promising candidate antigens for the development of subunit vaccines against brucellosis. This study was aimed to evaluate the protection of combination of recombinant HSA-L7/L12 fusion protein with LPS in Balb/c mouse. The recombinant HSA-L7/L12 fusion protein in Saccharomyces cerevisiae was expressed and purified by affinity chromatography column. LPS was extracted by n-butanol, purified by ultracentrifugation. BALB/c mouses were immunized in 9 groups with PBS, HSA, tHSA-L7/L12, L7/L12, LPS, LPS+ HSA, LPS+ tHSA-L7/L12, LPS+ L7/L12, B. abortus S19. ELISA, LTT tests and challenging two weeks after last injection were carried out. Bacterial count of spleen of immunized BALB/c mouse was done four weeks after challenging with virulent strain B. abortus 544. In ELISA test the specific antibodies of tHSA-L7/L12 exhibited a dominance of immunoglobulin IgG1 over IgG2a. LPS-HSA and tHSA-L7/L12 + LPS produced a significantly higher antibody titer than LPS alone and L7/L12+LPS (P < 0.05). The predominant IgG subtype for LPS and L7/L12+LPS were IgG3. However, tHSA-L7/L12+ LPS and LPS+ HAS elicited predominantly IgG1 and IgG3 subtypes. In addition, the tHSA-L7/L12 fusion protein and L7/L12 elicited a strong T-cell proliferative response upon restimulation in vitro with recombinant tHSA-L7/L12 and L7/L12, suggesting the induction of a cellular immunity response in vivo. However, there was no significant difference proliferative response in L7/L12 and tHSA-L7/L12 fusion protein (P > 0.05). The combination of tHSA-L7/L12 fusion protein with LPS and B. abortus S19 induce higher level of protection against challenge with the virulent strain B. abortus 544 in BALB/c mice than other groups (P = 0.005). The combination of tHSA-L7/L12 fusion protein with LPS had higher protective ability than LPS and fusion protein distinctly.

  2. Isolation of brucella abortus from two dogs in contact with bovine brucellosis.

    PubMed Central

    Prior, M G

    1976-01-01

    On a farm where several cattle were serologically positive for bovine brucellosis, three dogs were found to have titres greater than 400 i.u. to Brucella abortus. The titres persisted until the dogs were killed over two months later. Two male dogs were necropsied. B. abortus was isolated from the spleen of both dogs. While farm dogs are not thought to be a major reservoir of bovine brucellosis they may be considered as possible carriers in imfected herds and should be considered during the investigation and eradication of bovine brucellosis. PMID:826308

  3. Diverse Genetic Regulon of the Virulence-Associated Transcriptional Regulator MucR in Brucella abortus 2308

    PubMed Central

    Caswell, Clayton C.; Elhassanny, Ahmed E. M.; Planchin, Emilie E.; Roux, Christelle M.; Weeks-Gorospe, Jenni N.; Ficht, Thomas A.; Dunman, Paul M.

    2013-01-01

    The Ros-type regulator MucR is one of the few transcriptional regulators that have been linked to virulence in Brucella. Here, we show that a Brucella abortus in-frame mucR deletion strain exhibits a pronounced growth defect during in vitro cultivation and, more importantly, that the mucR mutant is attenuated in cultured macrophages and in mice. The genetic basis for the attenuation of Brucella mucR mutants has not been defined previously, but in the present study the genes regulated by MucR in B. abortus have been elucidated using microarray analysis and real-time reverse transcription-PCR (RT-PCR). In B. abortus 2308, MucR regulates a wide variety of genes whose products may function in establishing and maintaining cell envelope integrity, polysaccharide biosynthesis, iron homeostasis, genome plasticity, and transcriptional regulation. Particularly notable among the MucR-regulated genes identified is arsR6 (nolR), which encodes a transcriptional regulator previously linked to virulence in Brucella melitensis 16 M. Importantly, electrophoretic mobility shift assays (EMSAs) determined that a recombinant MucR protein binds directly to the promoter regions of several genes repressed by MucR (including arsR6 [nolR]), and in Brucella, as in other alphaproteobacteria, MucR binds to its own promoter to repress expression of the gene that encodes it. Overall, these studies have uncovered the diverse genetic regulon of MucR in Brucella, and in doing so this work has begun to define the MucR-controlled genetic circuitry whose misregulation contributes to the virulence defect of Brucella mucR mutants. PMID:23319565

  4. The bovine immune response to Brucella abortus. III. Preparation of antisera against a Brucella component precipitated by sera of some infected cattle.

    PubMed Central

    Stemshorn, B; Nielsen, K; Samagh, B

    1981-01-01

    Two methods are described for the partial purification of a high molecular weight, heat-resistant component (CO1) of sonicates of smooth and rough Brucella abortus which is precipitated by sera of some infected cattle. Method 1, a combination of gel filtration chromatography and polyacrylamide gel electrophoresis, was used to prepare CO1 from sonicates of a smooth field strain of B. abortus. Method 2, a combination of gel filtration chromatography and heat treatment, was used to obtain CO1, from sonicates of rough B. abortus strain 45/20. Rabbit antisera produced against CO1 prepared by either method contained only CO1 precipitins but were negative in standard agglutination and complement fixation tests conducted with whole cell antigens. Evidence is presented that CO1 is identical to Brucella antigen A2, and it is proposed that in future the designation A2 be employed. Images Fig. 1. Fig. 2. Fig. 3. Fig. 4. PMID:6791797

  5. Highly Sensitive Bacteriophage-Based Detection of Brucella abortus in Mixed Culture and Spiked Blood

    PubMed Central

    Sergueev, Kirill V.; Filippov, Andrey A.; Nikolich, Mikeljon P.

    2017-01-01

    For decades, bacteriophages (phages) have been used for Brucella species identification in the diagnosis and epidemiology of brucellosis. Traditional Brucella phage typing is a multi-day procedure including the isolation of a pure culture, a step that can take up to three weeks. In this study, we focused on the use of brucellaphages for sensitive detection of the pathogen in clinical and other complex samples, and developed an indirect method of Brucella detection using real-time quantitative PCR monitoring of brucellaphage DNA amplification via replication on live Brucella cells. This assay allowed the detection of single bacteria (down to 1 colony-forming unit per milliliter) within 72 h without DNA extraction and purification steps. The technique was equally efficient with Brucella abortus pure culture and with mixed cultures of B. abortus and α-proteobacterial near neighbors that can be misidentified as Brucella spp., Ochrobactrum anthropi and Afipia felis. The addition of a simple short sample preparation step enabled the indirect phage-based detection of B. abortus in spiked blood, with the same high sensitivity. This indirect phage-based detection assay enables the rapid and sensitive detection of live B. abortus in mixed cultures and in blood samples, and can potentially be applied for detection in other clinical samples and other complex sample types. PMID:28604602

  6. Comparison of Brucella abortus and Brucella melitensis infections of mice and their effect on acquired cellular resistance.

    PubMed Central

    Young, E J; Gomez, C I; Yawn, D H; Musher, D M

    1979-01-01

    By using mice infected with strains of Brucella abortus and Brucella melitensis we examined the histological responses to infection, the relationship of histology to persistence of organisms, and the relation of persistence of organisms to the acquisition of acquired cellular resistance (ACR). Infection with B. abortus resulted in well-formed granulomas in the livers, which persisted for more than 30 days. In contrast, infection with B. melitensis produced microabscesses in the livers which resolved before 30 days. The clearance of organisms from the tissues was also different. A total of 30 days after infection, large numbers of viable bacteria were recovered from the tissues of B. abortus-infected mice whereas bacteria were no longer recoverable from B. melitensis-infected animals. ACR to Listeria monocytogenes, another intracellular pathogen, persisted for more than 30 days in B. abortus-infected mice but waned rapidly in B. melitensis-infected animals. This disappearance of ACR due to B. melitensis paralleled the clearance of bacteria from the tissues. Images PMID:121113

  7. Epidemiology of Brucellosis and Genetic Diversity of Brucella abortus in Kazakhstan

    PubMed Central

    Shevtsova, Elena; Shevtsov, Alexandr; Mukanov, Kasim; Filipenko, Maxim; Kamalova, Dinara; Sytnik, Igor; Syzdykov, Marat; Kuznetsov, Andrey; Akhmetova, Assel; Zharova, Mira; Karibaev, Talgat; Tarlykov, Pavel; Ramanculov, Erlan

    2016-01-01

    Brucellosis is a major zoonotic infection in Kazakhstan. However, there is limited data on its incidence in humans and animals, and the genetic diversity of prevalent strains is virtually unstudied. Additionally, there is no detailed overview of Kazakhstan brucellosis control and eradication programs. Here, we analyzed brucellosis epidemiological data, and assessed the effectiveness of eradication strategies employed over the past 70 years to counteract this infection. We also conducted multiple loci variable-number tandem repeat analysis (MLVA) of Brucella abortus strains found in Kazakhstan. We analyzed official data on the incidence of animal brucellosis in Kazakhstan. The records span more than 70 years of anti-brucellosis campaigns, and contain a brief description of the applied control strategies, their effectiveness, and their impact on the incidence in humans. The MLVA-16 method was used to type 94 strains of B. abortus and serial passages of B. abortus 82, a strain used in vaccines. MLVA-8 and MLVA-11 analyses clustered strains into a total of four and seven genotypes, respectively; it is the first time that four of these genotypes have been described. MLVA-16 analysis divided strains into 28 distinct genotypes having genetic similarity coefficient that varies from 60 to100% and a Hunter & Gaston diversity index of 0.871. MST analysis reconstruction revealed clustering into "Kazakhstani-Chinese (Central Asian)", "European" and "American" lines. Detection of multiple genotypes in a single outbreak confirms that poorly controlled trade of livestock plays a crucial role in the spread of infection. Notably, the MLVA-16 profile of the B. abortus 82 strain was unique and did not change during 33 serial passages. MLVA genotyping may thus be useful for epidemiological monitoring of brucellosis, and for tracking the source(s) of infection. We suggest that countrywide application of MLVA genotyping would improve the control of brucellosis in Kazakhstan. PMID

  8. Epidemiology of Brucellosis and Genetic Diversity of Brucella abortus in Kazakhstan.

    PubMed

    Shevtsova, Elena; Shevtsov, Alexandr; Mukanov, Kasim; Filipenko, Maxim; Kamalova, Dinara; Sytnik, Igor; Syzdykov, Marat; Kuznetsov, Andrey; Akhmetova, Assel; Zharova, Mira; Karibaev, Talgat; Tarlykov, Pavel; Ramanculov, Erlan

    2016-01-01

    Brucellosis is a major zoonotic infection in Kazakhstan. However, there is limited data on its incidence in humans and animals, and the genetic diversity of prevalent strains is virtually unstudied. Additionally, there is no detailed overview of Kazakhstan brucellosis control and eradication programs. Here, we analyzed brucellosis epidemiological data, and assessed the effectiveness of eradication strategies employed over the past 70 years to counteract this infection. We also conducted multiple loci variable-number tandem repeat analysis (MLVA) of Brucella abortus strains found in Kazakhstan. We analyzed official data on the incidence of animal brucellosis in Kazakhstan. The records span more than 70 years of anti-brucellosis campaigns, and contain a brief description of the applied control strategies, their effectiveness, and their impact on the incidence in humans. The MLVA-16 method was used to type 94 strains of B. abortus and serial passages of B. abortus 82, a strain used in vaccines. MLVA-8 and MLVA-11 analyses clustered strains into a total of four and seven genotypes, respectively; it is the first time that four of these genotypes have been described. MLVA-16 analysis divided strains into 28 distinct genotypes having genetic similarity coefficient that varies from 60 to100% and a Hunter & Gaston diversity index of 0.871. MST analysis reconstruction revealed clustering into "Kazakhstani-Chinese (Central Asian)", "European" and "American" lines. Detection of multiple genotypes in a single outbreak confirms that poorly controlled trade of livestock plays a crucial role in the spread of infection. Notably, the MLVA-16 profile of the B. abortus 82 strain was unique and did not change during 33 serial passages. MLVA genotyping may thus be useful for epidemiological monitoring of brucellosis, and for tracking the source(s) of infection. We suggest that countrywide application of MLVA genotyping would improve the control of brucellosis in Kazakhstan.

  9. Variations in periplasmic loop interactions determine the pH-dependent activity of the hexameric urea transporter UreI from Helicobacter pylori: a molecular dynamics study.

    PubMed

    Cáceres-Delpiano, Javier; Teneb, Jaime; Mansilla, Rodrigo; García, Apolinaria; Salas-Burgos, Alexis

    2015-06-26

    Helicobacter pylori is an important factor in the development of diseases such as ulcer and gastric cancer. This bacterium uses a periplasmic transporter, UreI, to deliver urea to the intracelullar space, where later it is transformed into ammonia by the cytoplasmic enzyme urease to survive the acidic condition of the human stomach. The UreI transporter presents a pH-dependent activity, where this pH-dependence remains unknown at a structural level. Althought the existance of several protonable residues in the periplasmic loops are related to the pH-dependent activity, we find interesting to have a clear view of the conformational changes involved in this phenomena through a molecular dynamic study. Molecular dynamic simulations of the UreI transporter at three different pH conditions were performed, revealing two main pH-dependent conformations, which we present as the open and close states. We find that salt bridges between the periplasmic loops are crucial interactions that stabilize these conformations. Besides, a cooperative behaviour exists between the six subunits of the system that is necessary to fulfill the activity of this transporter. We found different pH-dependent conformations of the urea transporter UreI from Helicobacter pylori, which are related to salt-bridge interactions in the periplasmic regions. The behaviour of every channel in the system is not independent, given the existance of a cooperative behaviour through the formation of salt-bridges between the subunits of the hexameric system. We believe that our results will be related to the generation of new eradication therapies using this transporter as an attractive target, denoting that the knowledge of the possible pH-dependent conformations adopted for this transporter are important for the development of rational drug design approximations.

  10. Brucella abortus down-regulates MHC class II by the IL-6-dependent inhibition of CIITA through the downmodulation of IFN regulatory factor-1 (IRF-1).

    PubMed

    Velásquez, Lis N; Milillo, M Ayelén; Delpino, M Victoria; Trotta, Aldana; Fernández, Pablo; Pozner, Roberto G; Lang, Roland; Balboa, Luciana; Giambartolomei, Guillermo H; Barrionuevo, Paula

    2017-03-01

    Brucella abortus is an intracellular pathogen capable of surviving inside of macrophages. The success of B. abortus as a chronic pathogen relies on its ability to orchestrate different strategies to evade the adaptive CD4 + T cell responses that it elicits. Previously, we demonstrated that B. abortus inhibits the IFN-γ-induced surface expression of MHC class II (MHC-II) molecules on human monocytes, and this phenomenon correlated with a reduction in antigen presentation. However, the molecular mechanisms, whereby B. abortus is able to down-regulate the expression of MHC-II, remained to be elucidated. In this study, we demonstrated that B. abortus infection inhibits the IFN-γ-induced transcription of MHC-II, transactivator (CIITA) and MHC-II genes. Accordingly, we observed that the synthesis of MHC-II proteins was also diminished. B. abortus was not only able to reduce the expression of mature MHC-II, but it also inhibited the expression of invariant chain (Ii)-associated immature MHC-II molecules. Outer membrane protein 19 (Omp19), a prototypical B. abortus lipoprotein, diminished the expression of MHC-II and CIITA transcripts to the same extent as B. abortus infection. IL-6 contributes to these down-regulatory phenomena. In addition, B. abortus and its lipoproteins, through IL-6 secretion, induced the transcription of the negative regulators of IFN-γ signaling, suppressor of cytokine signaling (SOCS)-1 and -3, without interfering with STAT1 activation. Yet, B. abortus lipoproteins via IL-6 inhibit the expression of IFN regulatory factor 1 (IRF-1), a critical regulatory transcription factor for CIITA induction. Overall, these results indicate that B. abortus inhibits the expression of MHC-II molecules at very early points in their synthesis and in this way, may prevent recognition by T cells establishing a chronic infection. © Society for Leukocyte Biology.

  11. Comparative proteome analysis of laboratory grown Brucella abortus 2308 and Brucella melitensis 16M.

    PubMed

    Eschenbrenner, Michel; Horn, Troy A; Wagner, Mary Ann; Mujer, Cesar V; Miller-Scandle, Tabbi L; DelVecchio, Vito G

    2006-07-01

    Brucella species are pathogenic agents that cause brucellosis, a debilitating zoonotic disease that affects a large variety of domesticated animals and humans. Brucella melitensis and Brucella abortus are considered major health threats because of their highly infectious nature and worldwide occurrence. The availability of the annotated genomes for these two species has allowed a comparative proteomics study of laboratory grown B. melitensis 16M and B. abortus 2308 by two-dimensional (2-D) gel electrophoresis and peptide mass fingerprinting. Computer-assisted analysis of the different 2-D gel images of strains 16M and 2308 revealed significant quantitative and qualitative differences in their protein expression patterns. Proteins involved in membrane transport, particularly the high affinity amino acids binding proteins, and those involved in Sec-dependent secretion systems related to type IV and type V secretion systems, were differentially expressed. Differential expression of these proteins may be responsible for conferring specific host preference in the two strains 2308 and 16M.

  12. Electron tomography and cryo-SEM characterization reveals novel ultrastructural features of host-parasite interaction during Chlamydia abortus infection.

    PubMed

    Wilkat, M; Herdoiza, E; Forsbach-Birk, V; Walther, P; Essig, A

    2014-08-01

    Chlamydia (C.) abortus is a widely spread pathogen among ruminants that can be transmitted to women during pregnancy leading to severe systemic infection with consecutive abortion. As a member of the Chlamydiaceae, C. abortus shares the characteristic feature of an obligate intracellular biphasic developmental cycle with two morphological forms including elementary bodies (EBs) and reticulate bodies (RBs). In contrast to other chlamydial species, C. abortus ultrastructure has not been investigated yet. To do so, samples were fixed by high-pressure freezing and processed by different electron microscopic methods. Freeze-substituted samples were analysed by transmission electron microscopy, scanning transmission electron microscopical tomography and immuno-electron microscopy, and freeze-fractured samples were analysed by cryo-scanning electron microscopy. Here, we present three ultrastructural features of C. abortus that have not been reported up to now. Firstly, the morphological evidence that C. abortus is equipped with the type three secretion system. Secondly, the accumulation and even coating of whole inclusion bodies by membrane complexes consisting of multiple closely adjacent membranes which seems to be a C. abortus specific feature. Thirdly, the formation of small vesicles in the periplasmic space of RBs in the second half of the developmental cycle. Concerning the time point of their formation and the fact that they harbour chlamydial components, these vesicles might be morphological correlates of an intermediate step during the process of redifferentiation of RBs into EBs. As this feature has also been shown for C. trachomatis and C. pneumoniae, it might be a common characteristic of the family of Chlamydiaceae.

  13. Immunogencity of HSA-L7/L12 (Brucella abortus ribosomal protein) in an animal model.

    PubMed

    Pakzad, Iraj; Rezaee, Abbas; Rasaee, Mohammad Javad; Tabbaraee, Bahman; Delpisheh, Ali

    2009-03-01

    The immunogenic Brucella abortus ribosomal protein L7/L12 is a promising candidate antigen for the development of subunit vaccines against brucellosis. This study was aimed to evaluate the protection of recombinant Human Serum Albumin (HAS)-L7/L12 fusion protein in Balb/c mice. The amplified L7/L12 gene was cloned in pYHSA5 vector, pYHSA5-L7/L12 construct was transformed in Saccharomyces cerevisiae and the expressed protein from supernatant was purified by affinity chromatography. Balb/c mice were immunized in five groups by tHSA-L7/L12 fusion protein (group 1), Brucella abortus S19 (group 2), HSA (group 3), recombinant L7/L12 (group 4), PBS (group 5). ELISA to detect antibody production, LTT test to assess antigen specific lymphocyte response were conducted prior to virulent B. abortus strain 544 challenge two weeks after the last injection. Bacterial counts from spleens of immunized mice were done four weeks after challenge. In ELISA tests, the specific antibodies exhibited a dominance of immunoglobulin IgG1 over IgG2a. In addition, the tHSA-L7/L12 fusion protein and L7/L12 elicited a strong T-cell proliferative response upon restimulation in vitro with recombinant tHSA-L7/L12 and L7/L12, suggesting the induction of a cellular immunity response in vivo. However, there was no significant difference in proliferative response of L7/L12 and tHSA-L7/L12 fusion protein (p>0.05). The L7/L12 and tHSA-L7/L12 fusion protein vaccines could also induce significant protection against challenge with the virulent strain B. abortus 544 in Balb/c mice (p< or =0.05). The tHSA-L7/L12 fusion protein, similar to L7/L12 has the ability to induce antigen specific lymphocyte proliferation, stimulate humoral immunity and engender protection.

  14. [Multiplication of Brucella abortus and production of nitric oxide in two macrophage cell lines of different origin].

    PubMed

    Serafino, J; Conde, S; Zabal, O; Samartino, L

    2007-01-01

    Brucella abortus is a bacterium which causes abortions and infertility in cattle and undulant fever in humans. It multiplies intracellularly, evading the mechanisms of cellular death. Nitric oxide (NO) is important in the regulation of the immune response. In the present work, we studied the ability of three B. abortus strains to survive intracellularly in two macrophage cell lines. The bacterial multiplication in both cell lines was determined at two different times in UFC/ ml units. Moreover the inoculated cells were also observed under light-field and fluorescence microscopy stained with Giemsa and acridine orange, respectively. The stain of both cellular lines showed similar results with respect to the UFC/ml determination. The presence of B. abortus was confirmed by electronic microscopy. In both macrophage cell lines inoculated with the rough strain RB51, the multiplication diminished and the level of NO was higher, compared with cells inoculated with smooth strains (S19 and 2308). These results suggest that the absence of O-chain of LPS probably affects the intracellular growth of B. abortus.

  15. Identification and Characterization of a High-Affinity Choline Uptake System of Brucella abortus

    PubMed Central

    Herrmann, Claudia K.; Bukata, Lucas; Melli, Luciano; Marchesini, M. Ines; Caramelo, Julio J.

    2013-01-01

    Phosphatidylcholine (PC), a common phospholipid of the eukaryotic cell membrane, is present in the cell envelope of the intracellular pathogen Brucella abortus, the etiological agent of bovine brucellosis. In this pathogen, the biosynthesis of PC proceeds mainly through the phosphatidylcholine synthase pathway; hence, it relies on the presence of choline in the milieu. These observations imply that B. abortus encodes an as-yet-unknown choline uptake system. Taking advantage of the requirement of choline uptake for PC synthesis, we devised a method that allowed us to identify a homologue of ChoX, the high-affinity periplasmic binding protein of the ABC transporter ChoXWV. Disruption of the choX gene completely abrogated PC synthesis at low choline concentrations in the medium, thus indicating that it is a high-affinity transporter needed for PC synthesis via the PC synthase (PCS) pathway. However, the synthesis of PC was restored when the mutant was incubated in media with higher choline concentrations, suggesting the presence of an alternative low-affinity choline uptake activity. By means of a fluorescence-based equilibrium-binding assay and using the kinetics of radiolabeled choline uptake, we show that ChoX binds choline with an extremely high affinity, and we also demonstrate that its activity is inhibited by increasing choline concentrations. Cell infection assays indicate that ChoX activity is required during the first phase of B. abortus intracellular traffic, suggesting that choline concentrations in the early and intermediate Brucella-containing vacuoles are limited. Altogether, these results suggest that choline transport and PC synthesis are strictly regulated in B. abortus. PMID:23161032

  16. First isolation, identification, phenotypic and genotypic characterization of Brucella abortus biovar 3 from dairy cattle in Tanzania.

    PubMed

    Mathew, C; Stokstad, M; Johansen, T B; Klevar, S; Mdegela, R H; Mwamengele, G; Michel, P; Escobar, L; Fretin, D; Godfroid, J

    2015-07-21

    Brucellosis is a disease of worldwide public health and economic importance. Successful control is based on knowledge of epidemiology and strains present in an area. In developing countries, most investigations are based on serological assays. This study aimed at investigating a dairy herd experiencing abortions in order to establish within-herd seroprevalence to Brucella spp., identify, characterize Brucella strains by Multiple Loci Variable Number of Tandem Repeats Analysis (MLVA-VNTR) and investigate possible spillover to other species. The within-herd seroprevalence in cattle (n = 200) was 48 % (95 % CI 41-55), using an indirect ELISA, while the Rose Bengal Test (RBT) yielded lower prevalence (21.5 %; 95 % CI 16-27). Two sheep (n = 35) and one goat (n = 50) were seropositive using ELISA while none of the dogs (n = 6) was positive with the RBT. Three Brucella were isolated from an aborted fetus and associated membranes. Real time PCR (IS711), Bruce-ladder and classical biotyping classified the isolates as B. abortus biovar 3. MLVA-VNTR revealed two different but closely related genotypes. The isolates showed unique profiles, providing the first genotypic data from Tanzania. These genotypes were not related to B. abortus biovar 3 reference strain Tulya originally isolated from a human patient in Uganda in 1958, unlike the genotypes isolated and characterized recently in Kenya. High within-herd prevalence, isolation of the pathogen and abortion confirm that B. abortus is circulating in this herd with cattle as reservoir hosts. A low seroprevalence in sheep and goats suggests a spillover of B. abortus from cattle to small ruminants in the herd. This is the first isolation and characterization of B. abortus biovar 3 from a dairy cow with abortion in Tanzania. The origin of the Tanzanian genotypes remain elusive, although they seem to be related to genotypes found in Europe, Turkey and China but not related to B. abortus biovar 3 reference strain or genotypes from

  17. Detection of Leptospira spp. and Brucella abortus antibodies in free-living jaguars (Panthera onca) in two protected areas of northern Pantanal, Brazil.

    PubMed

    Onuma, Selma Samiko Miyazaki; Kantek, Daniel Luis Zanella; Crawshaw Júnior, Peter Gransden; Morato, Ronaldo Gonçalves; May-Júnior, Joares Adenilson; Morais, Zenaide Maria de; Ferreira Neto, José Soares; Aguiar, Daniel Moura de

    2015-01-01

    This study aimed to assess the exposure of free-living jaguars (Panthera onca) to Leptospira spp. and Brucella abortus in two conservation units in the Pantanal of Mato Grosso, Brazil. The presence of antibodies in blood samples of eleven jaguars was investigated using autochthonous antigens isolated in Brazil added to reference antigen collection applied to diagnosis of leptospirosis by Microscopic Agglutination Test (MAT). The Rose Bengal test was applied for B. abortus antibodies. Two (18.2%) jaguars were seroreactive for the Leptospira spp. antigen and the serovar considered as most infective in both animals was a Brazilian isolate of serovar Canicola (L01). All jaguars were seronegative for B. abortus. These data indicate that the inclusion of autochthonous antigens in serological studies can significantly increase the number of reactive animals, as well as modify the epidemiological profile of Leptospira spp. infection.

  18. PPARγ-mediated increase in glucose availability sustains chronic Brucella abortus infection in alternatively activated macrophages

    PubMed Central

    Xavier, Mariana N.; Winter, Maria G.; Spees, Alanna M.; den Hartigh, Andreas B.; Nguyen, Kim; Roux, Christelle M.; Silva, Teane M. A.; Atluri, Vidya L.; Kerrinnes, Tobias; Keestra, A. Marijke; Monack, Denise M.; Luciw, Paul A.; Eigenheer, Richard A.; Bäumler, Andreas J.; Santos, Renato L.; Tsolis, Renée M.

    2013-01-01

    SUMMARY Eradication of persistent intracellular bacterial pathogens with antibiotic therapy is often slow or incomplete. However, strategies to augment antibiotics are hampered by our poor understanding of the nutritional environment that sustains chronic infection. Here we show that the intracellular pathogen Brucella abortus survives and replicates preferentially in alternatively activated macrophages (AAM), which are more abundant during chronic infection. A metabolic shift induced by peroxisome proliferator activated receptor γ (PPARγ), which increases intracellular glucose availability, is identified as a causal mechanism promoting enhanced bacterial survival in AAM. Glucose uptake was crucial for increased replication of B. abortus in AAM, and chronic infection, as inactivation of the bacterial glucose transporter gluP reduced both intracellular survival in AAM and persistence in mice. Thus, a shift in intracellular nutrient availability induced by PPARγ promotes chronic persistence of B. abortus within AAM and targeting this pathway may aid in eradicating chronic infection. PMID:23954155

  19. Molecular typing of isolates obtained from aborted foetuses in Brucella-free Holstein dairy cattle herd after immunisation with Brucella abortus RB51 vaccine in Egypt.

    PubMed

    Wareth, Gamal; Melzer, Falk; Böttcher, Denny; El-Diasty, Mohamed; El-Beskawy, Mohamed; Rasheed, Nesma; Schmoock, Gernot; Roesler, Uwe; Sprague, Lisa D; Neubauer, Heinrich

    2016-12-01

    Bovine brucellosis is endemic in Egypt in spite of application of surveillance and control measures. An increase of abortions was reported in a Holstein dairy cattle herd with 600 animals in Damietta governorate in Egypt after immunisation with Brucella (B.) abortus RB51 vaccine. Twenty one (10.6%) of 197 vaccinated cows aborted after 3 months. All aborted cows had been tested seronegative for brucellosis in the past 3 years. B. abortus was isolated from four foetuses. Conventional biochemical and bacteriological identification and polymerase chain reaction (PCR) confirmed two B. abortus biovar (bv.) 1 smooth and two B. abortus rough strains. None of the B. abortus isolates were identified as RB51. Genotyping analysis by multiple locus of variable number tandem repeats analysis based on 16 markers (MLVA-16) revealed two different profiles with low genetic diversity. B. abortus bv1 was introduced in the herd and caused abortions. Copyright © 2016 Elsevier B.V. All rights reserved.

  20. Metallochaperone UreG serves as a new target for design of urease inhibitor: A novel strategy for development of antimicrobials.

    PubMed

    Yang, Xinming; Koohi-Moghadam, Mohamad; Wang, Runming; Chang, Yuen-Yan; Woo, Patrick C Y; Wang, Junwen; Li, Hongyan; Sun, Hongzhe

    2018-01-01

    Urease as a potential target of antimicrobial drugs has received considerable attention given its versatile roles in microbial infection. Development of effective urease inhibitors, however, is a significant challenge due to the deeply buried active site and highly specific substrate of a bacterial urease. Conventionally, urease inhibitors are designed by either targeting the active site or mimicking substrate of urease, which is not efficient. Up to now, only one effective inhibitor-acetohydroxamic acid (AHA)-is clinically available, but it has adverse side effects. Herein, we demonstrate that a clinically used drug, colloidal bismuth subcitrate, utilizes an unusual way to inhibit urease activity, i.e., disruption of urease maturation process via functional perturbation of a metallochaperone, UreG. Similar phenomena were also observed in various pathogenic bacteria, suggesting that UreG may serve as a general target for design of new types of urease inhibitors. Using Helicobacter pylori UreG as a showcase, by virtual screening combined with experimental validation, we show that two compounds targeting UreG also efficiently inhibited urease activity with inhibitory concentration (IC)50 values of micromolar level, resulting in attenuated virulence of the pathogen. We further demonstrate the efficacy of the compounds in a mammalian cell infection model. This study opens up a new opportunity for the design of more effective urease inhibitors and clearly indicates that metallochaperones involved in the maturation of important microbial metalloenzymes serve as new targets for devising a new type of antimicrobial drugs.

  1. Isolation, purification, and partial characterization of Brucella abortus matrix protein.

    PubMed Central

    Moriyon, I; Berman, D T

    1983-01-01

    Peptidoglycan sacculi with peptidoglycan-associated proteins were prepared from cell envelopes of Brucella abortus by extraction with sodium dodecyl sulfate (SDS) at 50 degrees C. On extraction of these preparations with SDS at 100 degrees C, a protein was obtained whose removal from peptidoglycan was confirmed by electron microscopy. Incubation of the 50 degrees C SDS-extracted cell envelopes with 50 mM MgCl2 in SDS-2-beta-mercaptoethanol at 37 degrees C also extracted the protein, along with lipopolysaccharide. At temperatures below 60 degrees C, the protein did not bind SDS strongly and had an apparent molecular weight greater than 92,000 in SDS-polyacrylamide gel electrophoresis. At higher temperatures, SDS bound strongly, and the apparent molecular weight was 38,000. Urea at 5 M did not alter the electrophoretic mobility of this 38,000-molecular-weight form. Immunoelectrophoresis in detergents with antisera to cell envelopes, carbohydrate staining of SDS-polyacrylamide gels, and production of anti-lipopolysaccharide antibodies by mice immunized with the purified protein indicated that lipopolysaccharide was present in free and protein-bound forms. Sequential gel filtration in SDS-EDTA and SDS-NaCl removed most lipopolysaccharide. After further purification by preparative SDS-polyacrylamide gel electrophoresis, a gas-liquid chromatographic analysis showed residual lipid tightly associated with the protein. The results suggested that the interactions between matrix proteins and other outer membrane components are stronger in B. abortus than in Escherichia coli, which was used as a control throughout. Images PMID:6401696

  2. DETECTION OF Leptospira spp. AND Brucella abortus ANTIBODIES IN FREE-LIVING JAGUARS (Panthera onca) IN TWO PROTECTED AREAS OF NORTHERN PANTANAL, BRAZIL

    PubMed Central

    ONUMA, Selma Samiko Miyazaki; KANTEK, Daniel Luis Zanella; CRAWSHAW, Peter Gransden; MORATO, Ronaldo Gonçalves; MAY-JÚNIOR, Joares Adenilson; de MORAIS, Zenaide Maria; FERREIRA, José Soares; de AGUIAR, Daniel Moura

    2015-01-01

     This study aimed to assess the exposure of free-living jaguars (Panthera onca) to Leptospira spp. and Brucella abortus in two conservation units in the Pantanal of Mato Grosso, Brazil. The presence of antibodies in blood samples of eleven jaguars was investigated using autochthonous antigens isolated in Brazil added to reference antigen collection applied to diagnosis of leptospirosis by Microscopic Agglutination Test (MAT). The Rose Bengal test was applied for B. abortus antibodies. Two (18.2%) jaguars were seroreactive for the Leptospira spp. antigen and the serovar considered as most infective in both animals was a Brazilian isolate of serovar Canicola (L01). All jaguars were seronegative for B. abortus. These data indicate that the inclusion of autochthonous antigens in serological studies can significantly increase the number of reactive animals, as well as modify the epidemiological profile of Leptospira spp. infection. PMID:25923900

  3. Real-time PCR Detection of Brucella Abortus: A Comparative Study of SYBR Green I, 5'-exonuclease, and Hybridization Probe Assays

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Newby, Deborah Trishelle; Hadfield, Ted; Roberto, Francisco Figueroa

    Real-time PCR provides a means of detecting and quantifying DNA targets by monitoring PCR product accumulation during cycling as indicated by increased fluorescence. A number of different approaches can be used to generate the fluorescence signal. Three approaches—SYBR Green I (a double-stranded DNA intercalating dye), 5'-exonuclease (enzymatically released fluors), and hybridization probes (fluorescence resonance energy transfer)—were evaluated for use in a real-time PCR assay to detect Brucella abortus. The three assays utilized the same amplification primers to produce an identical amplicon. This amplicon spans a region of the B. abortus genome that includes portions of the alkB gene and themore » IS711 insertion element. All three assays were of comparable sensitivity, providing a linear assay over 7 orders of magnitude (from 7.5 ng down to 7.5 fg). However, the greatest specificity was achieved with the hybridization probe assay.« less

  4. Dissemination and genetic diversity of chlamydial agents in Polish wildfowl: Isolation and molecular characterisation of avian Chlamydia abortus strains.

    PubMed

    Szymańska-Czerwińska, Monika; Mitura, Agata; Niemczuk, Krzysztof; Zaręba, Kinga; Jodełko, Agnieszka; Pluta, Aneta; Scharf, Sabine; Vitek, Bailey; Aaziz, Rachid; Vorimore, Fabien; Laroucau, Karine; Schnee, Christiane

    2017-01-01

    Wild birds are considered as a reservoir for avian chlamydiosis posing a potential infectious threat to domestic poultry and humans. Analysis of 894 cloacal or fecal swabs from free-living birds in Poland revealed an overall Chlamydiaceae prevalence of 14.8% (n = 132) with the highest prevalence noted in Anatidae (19.7%) and Corvidae (13.4%). Further testing conducted with species-specific real-time PCR showed that 65 samples (49.2%) were positive for C. psittaci whereas only one was positive for C. avium. To classify the non-identified chlamydial agents and to genotype the C. psittaci and C. avium-positive samples, specimens were subjected to ompA-PCR and sequencing (n = 83). The ompA-based NJ dendrogram revealed that only 23 out of 83 sequences were assigned to C. psittaci, in particular to four clades representing the previously described C. psittaci genotypes B, C, Mat116 and 1V. Whereas the 59 remaining sequences were assigned to two new clades named G1 and G2, each one including sequences recently obtained from chlamydiae detected in Swedish wetland birds. G1 (18 samples from Anatidae and Rallidae) grouped closely together with genotype 1V and in relative proximity to several C. abortus isolates, and G2 (41 samples from Anatidae and Corvidae) grouped closely to C. psittaci strains of the classical ABE cluster, Matt116 and M56. Finally, deep molecular analysis of four representative isolates of genotypes 1V, G1 and G2 based on 16S rRNA, IGS and partial 23S rRNA sequences as well as MLST clearly classify these isolates within the C. abortus species. Consequently, we propose an expansion of the C. abortus species to include not only the classical isolates of mammalian origin, but also avian isolates so far referred to as atypical C. psittaci or C. psittaci/C. abortus intermediates.

  5. Dissemination and genetic diversity of chlamydial agents in Polish wildfowl: Isolation and molecular characterisation of avian Chlamydia abortus strains

    PubMed Central

    Szymańska-Czerwińska, Monika; Mitura, Agata; Niemczuk, Krzysztof; Zaręba, Kinga; Jodełko, Agnieszka; Pluta, Aneta; Scharf, Sabine; Vitek, Bailey; Aaziz, Rachid; Vorimore, Fabien; Laroucau, Karine; Schnee, Christiane

    2017-01-01

    Wild birds are considered as a reservoir for avian chlamydiosis posing a potential infectious threat to domestic poultry and humans. Analysis of 894 cloacal or fecal swabs from free-living birds in Poland revealed an overall Chlamydiaceae prevalence of 14.8% (n = 132) with the highest prevalence noted in Anatidae (19.7%) and Corvidae (13.4%). Further testing conducted with species-specific real-time PCR showed that 65 samples (49.2%) were positive for C. psittaci whereas only one was positive for C. avium. To classify the non-identified chlamydial agents and to genotype the C. psittaci and C. avium-positive samples, specimens were subjected to ompA-PCR and sequencing (n = 83). The ompA-based NJ dendrogram revealed that only 23 out of 83 sequences were assigned to C. psittaci, in particular to four clades representing the previously described C. psittaci genotypes B, C, Mat116 and 1V. Whereas the 59 remaining sequences were assigned to two new clades named G1 and G2, each one including sequences recently obtained from chlamydiae detected in Swedish wetland birds. G1 (18 samples from Anatidae and Rallidae) grouped closely together with genotype 1V and in relative proximity to several C. abortus isolates, and G2 (41 samples from Anatidae and Corvidae) grouped closely to C. psittaci strains of the classical ABE cluster, Matt116 and M56. Finally, deep molecular analysis of four representative isolates of genotypes 1V, G1 and G2 based on 16S rRNA, IGS and partial 23S rRNA sequences as well as MLST clearly classify these isolates within the C. abortus species. Consequently, we propose an expansion of the C. abortus species to include not only the classical isolates of mammalian origin, but also avian isolates so far referred to as atypical C. psittaci or C. psittaci/C. abortus intermediates. PMID:28350846

  6. The bovine immune response to Brucella abortus. II. Elimination of some sporadic serological reactions by chelation of divalent cations.

    PubMed Central

    Nielsen, K; Samagh, B S; Speckmann, G; Stemshorn, B

    1979-01-01

    The standard agglutination tests for detecting antibody to Brucella abortus were modified by addition of chelating agents (EDTA and EGTA) to the antigens. Approximately 80% of "singleton" agglutination test reactions, negative on the diagnostic complement fixation test, obtained with cattle sera were eliminated while no decrease in titer was apparent when sera from B. abortus infected or vaccinated cattle were tested. PMID:121242

  7. Brucella abortus Synthesizes Phosphatidylcholine from Choline Provided by the Host

    PubMed Central

    Comerci, Diego J.; Altabe, Silvia; de Mendoza, Diego; Ugalde, Rodolfo A.

    2006-01-01

    The Brucella cell envelope is characterized by the presence of phosphatidylcholine (PC), a common phospholipid in eukaryotes that is rare in prokaryotes. Studies on the composition of Brucella abortus 2308 phospholipids revealed that the synthesis of PC depends on the presence of choline in the culture medium, suggesting that the methylation biosynthetic pathway is not functional. Phospholipid composition of pmtA and pcs mutants indicated that in Brucella, PC synthesis occurs exclusively via the phosphatidylcholine synthase pathway. Transformation of Escherichia coli with an expression vector containing the B. abortus pcs homologue was sufficient for PC synthesis upon induction with IPTG (isopropyl-β-d-thiogalactopyranoside), while no PC formation was detected when bacteria were transformed with a vector containing pmtA. These findings imply that Brucella depends on choline provided by the host cell to form PC. We could not detect any obvious associated phenotype in the PC-deficient strain under vegetative or intracellular growth conditions in macrophages. However, the pcs mutant strain displays a reproducible virulence defect in mice, which suggests that PC is necessary to sustain a chronic infection process. PMID:16484204

  8. Vaccination of elk (Cervus canadensis) with Brucella abortus strain RB51 overexpressing superoxide dismutase and glycosyltransferase genes does not induce adequate protection against experimental brucella abortus challenge

    USDA-ARS?s Scientific Manuscript database

    In recent years, elk (Cervus canadensis) have been implicated as the source of Brucella abortus infection for numerous cattle herds in the Greater Yellowstone Area (GYA). In the face of environmental and ecological changes on the landscape, the range of infected elk is expanding. Consequently, the d...

  9. Epidemiology of brucellosis in domestic animals caused by Brucella melitensis, Brucella suis and Brucella abortus.

    PubMed

    Díaz Aparicio, E

    2013-04-01

    Brucellosis is a disease that causes severe economic losses for livestock farms worldwide. Brucella melitensis, B. abortus and B. suis, which are transmitted between animals both vertically and horizontally, cause abortion and infertility in their primary natural hosts - goats and sheep (B. melitensis), cows (B. abortus) and sows (B. suis). Brucella spp. infect not only their preferred hosts but also other domestic and wild animal species, which in turn can act as reservoirs of the disease for other animal species and humans. Brucellosis is therefore considered to be a major zoonosis transmitted by direct contact with animals and/or their secretions, or by consuming milk and dairy products.

  10. Oral immunization of mice with gamma-irradiated Brucella neotomae induces protection against intraperitoneal and intranasal challenge with virulent B. abortus 2308.

    PubMed

    Dabral, Neha; Martha-Moreno-Lafont; Sriranganathan, Nammalwar; Vemulapalli, Ramesh

    2014-01-01

    Brucella spp. are Gram-negative, facultative intracellular coccobacilli that cause one of the most frequently encountered zoonosis worldwide. Humans naturally acquire infection through consumption of contaminated dairy and meat products and through direct exposure to aborted animal tissues and fluids. No vaccine against brucellosis is available for use in humans. In this study, we tested the ability of orally inoculated gamma-irradiated B. neotomae and B. abortus RB51 in a prime-boost immunization approach to induce antigen-specific humoral and cell mediated immunity and protection against challenge with virulent B. abortus 2308. Heterologous prime-boost vaccination with B. abortus RB51 and B. neotomae and homologous prime-boost vaccination of mice with B. neotomae led to the production of serum and mucosal antibodies specific to the smooth LPS. The elicited serum antibodies included the isotypes of IgM, IgG1, IgG2a, IgG2b and IgG3. All oral vaccination regimens induced antigen-specific CD4(+) and CD8(+) T cells capable of secreting IFN-γ and TNF-α. Upon intra-peritoneal challenge, mice vaccinated with B. neotomae showed the highest level of resistance against virulent B. abortus 2308 colonization in spleen and liver. Experiments with different doses of B. neotomae showed that all tested doses of 10(9), 10(10) and 10(11) CFU-equivalent conferred significant protection against the intra-peritoneal challenge. However, a dose of 10(11) CFU-equivalent of B. neotomae was required for affording protection against intranasal challenge as shown by the reduced bacterial colonization in spleens and lungs. Taken together, these results demonstrate the feasibility of using gamma-irradiated B. neotomae as an effective and safe oral vaccine to induce protection against respiratory and systemic infections with virulent Brucella.

  11. Development of a dual vaccine for prevention of Brucella abortus infection and Escherichia coli O157:H7 intestinal colonization.

    PubMed

    Iannino, Florencia; Herrmann, Claudia K; Roset, Mara S; Briones, Gabriel

    2015-05-05

    Zoonoses that affect human and animal health have an important economic impact. In the study now presented, a bivalent vaccine has been developed that has the potential for preventing the transmission from cattle to humans of two bacterial pathogens: Brucella abortus and Shiga toxin-producing Escherichia coli (STEC). A 66kDa chimeric antigen, composed by EspA, Intimin, Tir, and H7 flagellin (EITH7) from STEC, was constructed and expressed in B. abortus Δpgm vaccine strain (BabΔpgm). Mice orally immunized with BabΔpgm(EITH7) elicited an immune response with the induction of anti-EITH7 antibodies (IgA) that clears an intestinal infection of E. coli O157:H7 three times faster (t=4 days) than mice immunized with BabΔpgm carrier strain (t=12 days). As expected, mice immunized with BabΔpgm(EITH7) strain also elicited a protective immune response against B. abortus infection. A Brucella-based vaccine platform is described capable of eliciting a combined protective immune response against two bacterial pathogens with diverse lifestyles-the intracellular pathogen B. abortus and the intestinal extracellular pathogen STEC. Copyright © 2015 Elsevier Ltd. All rights reserved.

  12. Host-pathogen interactions in specific pathogen-free chickens following aerogenous infection with Chlamydia psittaci and Chlamydia abortus.

    PubMed

    Kalmar, Isabelle; Berndt, Angela; Yin, Lizi; Chiers, Koen; Sachse, Konrad; Vanrompay, Daisy

    2015-03-15

    Although Chlamydia (C.) psittaci infections are recognized as an important factor causing economic losses and impairing animal welfare in poultry production, the specific mechanisms leading to severe clinical outcomes are poorly understood. In the present study, we comparatively investigated pathology and host immune response, as well as systemic dissemination and expression of essential chlamydial genes in the course of experimental aerogeneous infection with C. psittaci and the closely related C. abortus, respectively, in specific pathogen-free chicks. Clinical signs appeared sooner and were more severe in the C. psittaci-infected group. Compared to C. abortus infection, more intense systemic dissemination of C. psittaci correlated with higher and faster infiltration of immune cells, as well as more macroscopic lesions and epithelial pathology, such as hyperplasia and erosion. In thoracic air sac tissue, mRNA expression of immunologically relevant factors, such as IFN-γ, IL-1β, IL-6, IL-17, IL-22, LITAF and iNOS was significantly stronger up-regulated in C. psittaci- than in C. abortus-infected birds between 3 and 14 days post-infection. Likewise, transcription rates of the chlamydial genes groEL, cpaf and ftsW were consistently higher in C. psittaci during the acute phase. These findings illustrate that the stronger replication of C. psittaci in its natural host also evoked a more intense immune response than in the case of C. abortus infection. Copyright © 2015 Elsevier B.V. All rights reserved.

  13. [Detection of Chlamydia abortus in bovine reproductive losses in the province of La Pampa, Argentina].

    PubMed

    Rojas, María Del C; Fort, Marcelo; Bettermann, Simone; Entrocassi, Carolina; Costamagna, Sixto R; Sachse, Konrad; Rodríguez Fermepin, Marcelo

    2018-01-16

    Reproductive losses linked to an infectious etiology in bovine cattle are a major economic concern worldwide. In Argentina, more than 50% of abortion cases have unknown causes. Species belonging to Chlamydiaceae family are frequent etiologic agents of abortion around the world; however, there is yet no information on their prevalence in Argentina. The objective of this work was to identify Chlamydia spp., and particularly C. abortus in reproductive losses from bovine cattle in La Pampa, Argentina. Real time PCR targeting Chlamydiaceae-specific DNA fragments was performed on 251 samples obtained from bovine abortions and stillborns, and ArrayTube was used for species identification on positive samples. Chlamydiaceae DNA was detected in 12 samples of aborted fetuses (4.78%), 83.33% (10/12) accounting for abortions and 16.66% (2/12) for stillborns. C. abortus was detected by ArrayTube in 5 cases (1.99% of all samples, and 41.67% of Chlamydiaceae positive samples). This study shows the first detection of Chlamydiaceae and C. abortus DNA on reproductive losses of bovine cattle in Argentina, and the described prevalence value (4.78%) should be taken as baseline value due to the type of samples analyzed. Detection of genetic material from Chlamydiaceae not matching any of the studied species could be due to intraspecies variants or local species not yet described. Further research on Chlamydia infections in bovine cattle in Argentina is imperative to describe their range, to analyze their economic and zoonotic implications and to make recommendations about prevention and control measures. Copyright © 2017 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.

  14. Dietary Pyridoxine Controls Efficacy of Vitamin B6-Auxotrophic Tuberculosis Vaccine Bacillus Calmette-Guérin ΔureC::hly Δpdx1 in Mice

    PubMed Central

    Vogelzang, Alexis; Schuerer, Stefanie; Lazar, Doris; Kaiser, Peggy

    2014-01-01

    ABSTRACT The only tuberculosis (TB) vaccine in use today, bacillus Calmette-Guérin (BCG), provides insufficient protection and can cause adverse events in immunocompromised individuals, such as BCGosis in HIV+ newborns. We previously reported improved preclinical efficacy and safety of the recombinant vaccine candidate BCG ΔureC::hly, which secretes the pore-forming listeriolysin O of Listeria monocytogenes. Here, we evaluate a second-generation construct, BCG ΔureC::hly Δpdx1, which is deficient in pyridoxine synthase, an enzyme that is required for biosynthesis of the essential cofactor vitamin B6. This candidate was auxotrophic for vitamin B6 in a concentration-dependent manner, as was its survival in vivo. BCG ΔureC::hly Δpdx1 showed markedly restricted dissemination in subcutaneously vaccinated mice, which was ameliorated by dietary supplementation with vitamin B6. The construct was safer in severe combined immunodeficiency mice than the parental BCG ΔureC::hly. A prompt innate immune response to vaccination, measured by secretion of interleukin-6, granulocyte colony-stimulating factor, keratinocyte cytokine, and macrophage inflammatory protein-1α, remained independent of vitamin B6 administration, while acquired immunity, notably stimulation of antigen-specific CD4 T cells, B cells, and memory T cells, was contingent on vitamin B6 administration. The early protection provided by BCG ΔureC::hly Δpdx1 in a murine Mycobacterium tuberculosis aerosol challenge model consistently depended on vitamin B6 supplementation. Prime-boost vaccination increased protection against the canonical M. tuberculosis H37Rv laboratory strain and a clinical isolate of the Beijing/W lineage. We demonstrate that the efficacy of a profoundly attenuated recombinant BCG vaccine construct can be modulated by external administration of a small molecule. This principle fosters the development of safer vaccines required for immunocompromised individuals, notably HIV+ infants. PMID

  15. Intracellularly Induced Cyclophilins Play an Important Role in Stress Adaptation and Virulence of Brucella abortus

    PubMed Central

    García Fernández, Lucía; DelVecchio, Vito G.; Briones, Gabriel

    2013-01-01

    Brucella is an intracellular bacterial pathogen that causes the worldwide zoonotic disease brucellosis. Brucella virulence relies on its ability to transition to an intracellular lifestyle within host cells. Thus, this pathogen must sense its intracellular localization and then reprogram gene expression for survival within the host cell. A comparative proteomic investigation was performed to identify differentially expressed proteins potentially relevant for Brucella intracellular adaptation. Two proteins identified as cyclophilins (CypA and CypB) were overexpressed in the intracellular environment of the host cell in comparison to laboratory-grown Brucella. To define the potential role of cyclophilins in Brucella virulence, a double-deletion mutant was constructed and its resulting phenotype was characterized. The Brucella abortus ΔcypAB mutant displayed increased sensitivity to environmental stressors, such as oxidative stress, pH, and detergents. In addition, the B. abortus ΔcypAB mutant strain had a reduced growth rate at lower temperature, a phenotype associated with defective expression of cyclophilins in other microorganisms. The B. abortus ΔcypAB mutant also displays reduced virulence in BALB/c mice and defective intracellular survival in HeLa cells. These findings suggest that cyclophilins are important for Brucella virulence and survival in the host cells. PMID:23230297

  16. Novel vector vaccine against Brucella abortus based on influenza A viruses expressing Brucella L7/L12 or Omp16 proteins: evaluation of protection in pregnant heifers.

    PubMed

    Tabynov, Kaissar; Yespembetov, Bolat; Sansyzbay, Abylai

    2014-10-14

    The present study provides the first information about the protection of a novel influenza viral vector vaccine expressing the Brucella proteins ribosomal L7/L12 or Omp16 containing the adjuvant Montanide Gel01 in pregnant heifers. Immunization of pregnant heifers was conducted via the conjunctival (n=10) or subcutaneous (n=10) route using cross prime and booster vaccination schedules at an interval of 28 days. The vector vaccine was evaluated in comparison with positive control groups vaccinated with Brucella abortus S19 (n=10) or B. abortus RB51 (n=10) and a negative (PBS+Montanide Gel01; n=10) control group. Via both the conjunctival or subcutaneous route, evaluation of protectiveness against abortion, effectiveness of vaccination and index of infection (in heifers and their fetuses or calves) demonstrated the vector vaccine provided good protection against B. abortus 544 infection compared to the negative control group (PBS+Montanide Gel01) and comparable protection to commercial vaccines B. abortus S19 or B. abortus RB51. Copyright © 2014 Elsevier Ltd. All rights reserved.

  17. Herd-level prevalence and associated risk factors for Toxoplasma gondii, Neospora caninum, Chlamydia abortus and bovine viral diarrhoea virus in commercial dairy and beef cattle in eastern, northern and northeastern China.

    PubMed

    Sun, Wu-Wen; Meng, Qing-Feng; Cong, Wei; Shan, Xiao-Feng; Wang, Chun-Feng; Qian, Ai-Dong

    2015-11-01

    Although the seroprevalence of Toxoplasma gondii, Neospora caninum, Chlamydia abortus and bovine viral diarrhea virus infection in cattle have been reported in some areas in China, most of them were conducted with small number of cattle samples and very limited districts and neglected the assessment of herd management factors associated with herd-level prevalence of these pathogen infections. Thus, from September 2013 to December 2014, a large-scale seroprevalence study was conducted to determine the animal-level and herd-level seroprevalence and identify herd-level risk factors associated with these pathogen infections in 4487 cattle from 134 herds in five provinces (Heilongjiang, Jilin, Liaoning, Shandong, Hebei) and Inner Mongolia Autonomous Region of China. At animal level, the true prevalence of antibodies against T. gondii, N. caninum, C. abortus and bovine viral diarrhoea virus (BVDV) was 10.48, 17.14, 11.92 and 50.10%, respectively. At herd level, the true prevalence of antibodies against T. gondii, N. caninum, C. abortus and BVDV was 27.16, 29.10, 37.31 and 40.30%, respectively. Multivariate analysis of these characteristics showed that source of water and presence of felids were significantly associated with T. gondii infection in the studied cattle herds. Source of water was significantly associated with N. caninum infection in the studied cattle herds. While herd size and management system were significantly associated with BVDV infection in the studied cattle herds, this is the first report of herd-level prevalence and associated risk factors of T. gondii, N. caninum, C. abortus and BVDV infection in cattle in China.

  18. Purification and properties of Cu-Zn superoxide dismutase extracted from Brucella abortus strain 19

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Tabatabai, L.B.

    Recent work showed that a recombinant 20 kDa protein from Brucella abortus expressed in E. coli is a Cu-Zn superoxide dismutase (SOD). Western blot and ELISA results indicated that cattle with brucellosis have antibody to SOD. Here the authors report the purification and properties of the native B. abortus Cu-Zn SOD. SOD was extracted from methanol-killed Brucella abortus strain 19 with 0.1 M sodium citrate-1.0 M sodium chloride solution. The extract was dialyzed and protein precipitated by ammonium sulfate at 70-100% saturation was collected. The SOD was purified by HPLC anion exchange chromatography. SOD activity was assayed with a coupledmore » enzyme assay using xanthine oxidase-cytochrome C reduction assay. The authors determined that the Brucella SOD is present in two molecular forms both inhibitable with KCN with Ki's of 0.32 mM and 4.98 mM, respectively. No other form of SOD was identified in the extract. Polyclonal antibody to SOD and polyclonal antibody to SOD synthetic peptide residues 134-143 inhibited SOD activity by 50% and 13%, respectively. Both SOD and the synthetic peptide inhibited binding of anti-SOD antibody to SOD by 60% and 20%, respectively. Based on these results the SOD and its amphipathic peptide will be considered as candidates for the design of synthetic multiple peptide vaccines and diagnostic reagents for bovine brucellosis.« less

  19. Relationship between ureB Sequence Diversity, Urease Activity and Genotypic Variations of Different Helicobacter pylori Strains in Patients with Gastric Disorders.

    PubMed

    Ghalehnoei, Hossein; Ahmadzadeh, Alireza; Farzi, Nastaran; Alebouyeh, Masoud; Aghdaei, Hamid Asadzadeh; Azimzadeh, Pendram; Molaei, Mahsa; Zali, Mohammad Reza

    2016-01-01

    Association of the severity of Helicobacter pylori induced diseases with virulence entity of the colonized strains was proven in some studies. Urease has been demonstrated as a potent virulence factor for H. pylori. The main aim of this study was investigation of the relationships of ureB sequence diversity, urease activity and virulence genotypes of different H. pylori strains with histopathological changes of gastric tissue in infected patients suffering from different gastric disorders. Analysis of the virulence genotypes in the isolated strains indicated significant associations between the presence of severe active gastritis and cagA+ (P = 0.039) or cagA/iceA1 genotypes (P = 0.026), and intestinal metaplasia and vacA m1 (P = 0.008) or vacA s1/m2 (P = 0.001) genotypes. Our results showed a 2.4-fold increased risk of peptic ulcer (95% CI: 0.483-11.93), compared with gastritis, in the infected patients who had dupA positive strains; however this association was not statistically significant. The results of urease activity showed a significant mean difference between the isolated strains from patients with PUD and NUD (P = 0.034). This activity was relatively higher among patients with intestinal metaplasia. Also a significant association was found between the lack of cagA and increased urease activity among the isolated strains (P = 0.036). While the greatest sequence variation of ureB was detected in a strain from a patient with intestinal metaplasia, the sole determined amino acid change in UreB sequence (Ala201Thr, 30%), showed no influence on urease activity. In conclusion, the supposed role of H. pylori urease to form peptic ulcer and advancing of intestinal metaplasia was postulated in this study. Higher urease activity in the colonizing H. pylori strains that present specific virulence factors was indicated as a risk factor for promotion of histopathological changes of gastric tissue that advance gastric malignancy.

  20. Brucella abortus Induces the Premature Death of Human Neutrophils through the Action of Its Lipopolysaccharide

    PubMed Central

    Barquero-Calvo, Elías; Mora-Cartín, Ricardo; Arce-Gorvel, Vilma; de Diego, Juana L.; Chacón-Díaz, Carlos; Chaves-Olarte, Esteban; Guzmán-Verri, Caterina; Buret, Andre G.; Gorvel, Jean-Pierre; Moreno, Edgardo

    2015-01-01

    Most bacterial infections induce the activation of polymorphonuclear neutrophils (PMNs), enhance their microbicidal function, and promote the survival of these leukocytes for protracted periods of time. Brucella abortus is a stealthy pathogen that evades innate immunity, barely activates PMNs, and resists the killing mechanisms of these phagocytes. Intriguing clinical signs observed during brucellosis are the low numbers of Brucella infected PMNs in the target organs and neutropenia in a proportion of the patients; features that deserve further attention. Here we demonstrate that B. abortus prematurely kills human PMNs in a dose-dependent and cell-specific manner. Death of PMNs is concomitant with the intracellular Brucella lipopolysaccharide (Br-LPS) release within vacuoles. This molecule and its lipid A reproduce the premature cell death of PMNs, a phenomenon associated to the low production of proinflammatory cytokines. Blocking of CD14 but not TLR4 prevents the Br-LPS-induced cell death. The PMNs cell death departs from necrosis, NETosis and classical apoptosis. The mechanism of PMN cell death is linked to the activation of NADPH-oxidase and a modest but steadily increase of ROS mediators. These effectors generate DNA damage, recruitments of check point kinase 1, caspases 5 and to minor extent of caspase 4, RIP1 and Ca++ release. The production of IL-1β by PMNs was barely stimulated by B. abortus infection or Br-LPS treatment. Likewise, inhibition of caspase 1 did not hamper the Br-LPS induced PMN cell death, suggesting that the inflammasome pathway was not involved. Although activation of caspases 8 and 9 was observed, they did not seem to participate in the initial triggering mechanisms, since inhibition of these caspases scarcely blocked PMN cell death. These findings suggest a mechanism for neutropenia in chronic brucellosis and reveal a novel Brucella-host cross-talk through which B. abortus is able to hinder the innate function of PMN. PMID:25946018

  1. Immunotoxic effect of thiamethoxam in immunized mice with Brucella abortus cultural filtrate antigen

    PubMed Central

    Salema, L. H.; Alwan, M. J.; Yousif, Afaf Abdulrahman

    2016-01-01

    Aim: This study was planned for determination the toxic effect of thiamethoxam (TMX) in immunized mice with Brucella abortus culture filtrate antigen (CFBAgs) (as a vaccine) and its role of TMX on decrease activity of B. abortus antigen on eliciting of humoral and cellular immunity. Materials and Methods: To achieve these goals 60 female mice were used, 7-8 weeks age, they were divided equally into three groups (20 in each group) and treated as follows: 1st group: Mice were immunized with CFBAgs intraperitoneally in two doses, 2 weeks intervals with (protein concentration 2 mg\\ml), 2nd group: Mice immunized as in the 1st group and was administrated orally with 1/10 lethal dose 50% of TMX (83.7 mg/kg B.W.) for 4 weeks daily, 3rd group was administrated orally with 0.3 ml normal saline served as a control group. At day 28 post immunization (PI) delayed type hypersensitivity (skin test) was done, and serum samples were collected at day 30 (PI) for detection of passive hemagglutination test (PHA); interferon gamma (IFN-γ) which was done by enzyme-linked immunosorbent assay test in addition to phagocytes assay. Results: The results of skin test post injection with soluble antigen of B. abortus intradermally showed a high significantly mean values at p≤0.05 of footpad skin thickness in the 1st group of mice which recorded (0.51±0.002 mm) as compared with the 2nd group of mice which showed (0.08±0.002 mm) after 24 h; the mean values of skin thickness were declined in the 1st mice (0.46±0.002) and 2nd mice (0.070±0.001) at 48 h; control group showed a negative results. These results were agreed with results of serum levels of IFN-γ (pg/ml) that showed that a significant increase the vaccinated 1st group (406.36±1.52), than those values in the 2nd group (151.61±0.89) and negative result in 3rd group (46.47±0.60), in addition to results of PHA test which showed a significant increase in antibody titer in the 1st group (139±12.16) with low level of serum antibody

  2. Recombinant Brucella abortus gene expressing immunogenic protein

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Mayfield, J.E.; Tabatabai, L.B.

    This patent describes a synthetic recombinant DNA molecule containing a DNA sequence. It comprises a gene of Brucella abortus encoding an immunogenic protein having a molecular weight of approximately 31,000 daltons as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis under denaturing conditions, the protein having an isoelectric point around 4.9, and containing a twenty-five amino acid sequence from its amino terminal end consisting of Gln-Ala-Pro-Thr-Phe-Phe-Arg-Ile-Gly-Thr-Gly-Gly-Thr-Ala-Gly-Thr-Tyr-Tyr-Pro-Ile-Gly-Gly-Leu-Ile-Ala, wherein Gln, Ala, Pro, Thr, Phe, Arg, Ile, Gly, Tyr, and Leu, respectively, represent glutamine, alanine, proline, threonine, phenylalanine, arginine, isolecuine, glycine, tyrosine, and leucine.

  3. Dietary pyridoxine controls efficacy of vitamin B6-auxotrophic tuberculosis vaccine bacillus Calmette-Guérin ΔureC::hly Δpdx1 in mice.

    PubMed

    Gengenbacher, Martin; Vogelzang, Alexis; Schuerer, Stefanie; Lazar, Doris; Kaiser, Peggy; Kaufmann, Stefan H E

    2014-06-03

    The only tuberculosis (TB) vaccine in use today, bacillus Calmette-Guérin (BCG), provides insufficient protection and can cause adverse events in immunocompromised individuals, such as BCGosis in HIV(+) newborns. We previously reported improved preclinical efficacy and safety of the recombinant vaccine candidate BCG ΔureC::hly, which secretes the pore-forming listeriolysin O of Listeria monocytogenes. Here, we evaluate a second-generation construct, BCG ΔureC::hly Δpdx1, which is deficient in pyridoxine synthase, an enzyme that is required for biosynthesis of the essential cofactor vitamin B6. This candidate was auxotrophic for vitamin B6 in a concentration-dependent manner, as was its survival in vivo. BCG ΔureC::hly Δpdx1 showed markedly restricted dissemination in subcutaneously vaccinated mice, which was ameliorated by dietary supplementation with vitamin B6. The construct was safer in severe combined immunodeficiency mice than the parental BCG ΔureC::hly. A prompt innate immune response to vaccination, measured by secretion of interleukin-6, granulocyte colony-stimulating factor, keratinocyte cytokine, and macrophage inflammatory protein-1α, remained independent of vitamin B6 administration, while acquired immunity, notably stimulation of antigen-specific CD4 T cells, B cells, and memory T cells, was contingent on vitamin B6 administration. The early protection provided by BCG ΔureC::hly Δpdx1 in a murine Mycobacterium tuberculosis aerosol challenge model consistently depended on vitamin B6 supplementation. Prime-boost vaccination increased protection against the canonical M. tuberculosis H37Rv laboratory strain and a clinical isolate of the Beijing/W lineage. We demonstrate that the efficacy of a profoundly attenuated recombinant BCG vaccine construct can be modulated by external administration of a small molecule. This principle fosters the development of safer vaccines required for immunocompromised individuals, notably HIV(+) infants

  4. Seroprevalence for Coxiella burnetii, Francisella tularensis, Brucella abortus and Brucella melitensis in Austrian adults: a cross-sectional survey among military personnel and civilians.

    PubMed

    Tobudic, Selma; Nedomansky, Klara; Poeppl, Wolfgang; Müller, Maria; Faas, Angelus; Mooseder, Gerhard; Allerberger, Franz; Stanek, Gerold; Burgmann, Heinz

    2014-04-01

    The prevalence of Coxiella burnetii, Francisella tularensis, Brucella abortus, and Brucella melitensis infections in Austria and the exposure risk of military personnel were assessed in an exploratory nationwide cross-sectional seroprevalence survey in 526 healthy adult individuals, 222 of which were soldiers and 304 were civilians. Screening for IgA/IgG antibodies to C. burnetii (Phase I) and IgG/IgM antibodies to C. burnetii (Phase II), and to F. tularensis was done with commercial enzyme-linked immunosorbent assays. To detect antibodies against B. abortus and B. melitensis, an in-house complement fixation test was used. Overall, 11 individuals (2.0%) showed antibodies to C. burnetii, 3 individuals (0.5%) were seropositive for F. tularensis, and one (0.3%) individual was borderline positive. All individuals positive or borderline for F. tularensis tested negative for antibodies against C. burnetii. All individuals tested negative for antibodies against B. melitensis/B. abortus. There were no significant differences between the seroprevalence of C. burnetii and F. tularensis among military personnel and civilians. Our data demonstrate serological evidence of a low rate of exposure to C. burnetii and F. tularensis among the Austrian adult population and military personnel. Copyright © 2014 Elsevier GmbH. All rights reserved.

  5. TLR2 and TLR4 signaling pathways are required for recombinant Brucella abortus BCSP31-induced cytokine production, functional upregulation of mouse macrophages, and the Th1 immune response in vivo and in vitro.

    PubMed

    Li, Jia-Yun; Liu, Yuan; Gao, Xiao-Xue; Gao, Xiang; Cai, Hong

    2014-09-01

    Brucella abortus is a zoonotic Gram-negative pathogen that causes brucelosis in ruminants and humans. Toll-like receptors (TLRs) recognize Brucella abortus and initiate antigen-presenting cell activities that affect both innate and adaptive immunity. In this study, we focused on recombinant Brucella cell-surface protein 31 (rBCSP31) to determine its effects on mouse macrophages. Our results demonstrated that rBCSP31 induced TNF-α, IL-6 and IL-12p40 production, which depended on the activation of mitogen-activated protein kinases (MAPKs) by stimulating the rapid phosphorylation of p38 and JNK and the activation of transcription factor NF-κB in macrophages. In addition, continuous exposure (>24 h) of RAW264.7 cells to rBCSP31 significantly enhanced IFN-γ-induced expression of MHC-II and the ability to present rBCSP31 peptide to CD4(+) T cells. Furthermore, we found that rBCSP31 could interact with both TLR2 and TLR4. The rBCSP31-induced cytokine production by macrophages from TLR2(-/-) and TLR4(-/-) mice was lower than that from C57BL/6 macrophages, and the activation of NF-κB and MAPKs was attenuated in macrophages from TLR2(-/-) and TLR4(-/-) mice. In addition, CD4(+) T cells from C57BL/6 mice immunized with rBCSP31 produced higher levels of IFN-γ and IL-2 compared with CD4(+) T cells from TLR2(-/-) and TLR4(-/-) mice. Macrophages from immunized C57BL/6 mice produced higher levels of IL-12p40 than those from TLR2(-/-) and TLR4(-/-) mice. Furthermore, immunization with rBCSP31 provided better protection in C57BL/6 mice than in TLR2(-/-) and TLR4(-/-) mice after B. abortus 2308 challenge. These results indicate that rBCSP31 is a TLR2 and TLR4 agonist that induces cytokine production, upregulates macrophage function and induces the Th1 immune response.

  6. Profiling Antibody Responses to Infections by Chlamydia abortus Enables Identification of Potential Virulence Factors and Candidates for Serodiagnosis

    PubMed Central

    Forsbach-Birk, Vera; Foddis, Corinna; Simnacher, Ulrike; Wilkat, Max; Longbottom, David; Walder, Gernot; Benesch, Christiane; Ganter, Martin; Sachse, Konrad; Essig, Andreas

    2013-01-01

    Enzootic abortion of ewes (EAE) due to infection with the obligate intracellular pathogen Chlamydia (C.) abortus is an important zoonosis leading to considerable economic loss to agriculture worldwide. The pathogen can be transmitted to humans and may lead to serious infection in pregnant women. Knowledge about epidemiology, clinical course and transmission to humans is hampered by the lack of reliable diagnostic tools. Immunoreactive proteins, which are expressed in infected animals and humans, may serve as novel candidates for diagnostic marker proteins and represent putative virulence factors. In order to broaden the spectrum of immunogenic C. abortus proteins we applied 2D immunoblot analysis and screening of an expression library using human and animal sera. We have identified 48 immunoreactive proteins representing potential diagnostic markers and also putative virulence factors, such as CAB080 (homologue of the “macrophage infectivity potentiator”, MIP), CAB167 (homologue of the “translocated actin recruitment protein”, TARP), CAB712 (homologue of the “chlamydial protease-like activity factor”, CPAF), CAB776 (homologue of the “Polymorphic membrane protein D”, PmpD), and the “hypothetical proteins” CAB063, CAB408 and CAB821, which are predicted to be type III secreted. We selected two putative virulence factors for further characterization, i.e. CAB080 (cMIP) and CAB063, and studied their expression profiles at transcript and protein levels. Analysis of the subcellular localization of both proteins throughout the developmental cycle revealed CAB063 being the first C. abortus protein shown to be translocated to the host cell nucleus. PMID:24260366

  7. Bactericidal Action of Fresh Rabbit Blood Against Brucella abortus

    PubMed Central

    Joos, Richard W.; Hall, Wendell H.

    1968-01-01

    A photometric method was used to measure the bactericidal kinetics for Brucella abortus of freshly drawn rabbit blood during the time before clotting. This antibrucellar activity varied between rabbits in different immunologic states. Nonimmunized rabbits had moderate bactericidal activity after a lag of about 2 min. The blood of some immunized rabbits gave an immediate and strong kill, but in certain other immunized rabbits, especially when hyperimmunized, the bactericidal activity was inhibited. It appeared that serum bactericidins and complement are sometimes as active in unclotted blood as they are in serum. However, this bactericidal activity can be either increased or neutralized by immunization. The prozone bactericidal inhibition phenomenon (Neisser-Wechsberg) found in immune serum may, in fact, reflect inhibition taking place in vivo. Inhibition of the bactericidal activity in blood can contribute to the persistence of chronic infections and individual variations in resistance. PMID:4971893

  8. The Structure of Urease Activation Complexes Examined by Flexibility Analysis, Mutagenesis, and Small-Angle X-Ray Scattering

    PubMed Central

    Quiroz-Valenzuela, Soledad; Sukuru, Sai Chetan K.; Hausinger, Robert P.; Kuhn, Leslie A.; Heller, William T.

    2008-01-01

    Conformational changes of Klebsiella aerogenes urease apoprotein (UreABC)3 induced upon binding of the UreD and UreF accessory proteins were examined by a combination of flexibility analysis, mutagenesis, and small-angle x-ray scattering (SAXS). ProFlex analysis of urease provided evidence that the major domain of UreB can move in a hinge-like motion to account for prior chemical cross-linking results. Rigidification of the UreB hinge region, accomplished through a G11P mutation, reduced the extent of urease activation, in part by decreasing the nickel content of the mutant enzyme, and by sequestering a portion of the urease apoprotein in a novel activation complex that includes all of the accessory proteins. SAXS analyses of urease, (UreABC-UreD)3, and (UreABC-UreDF)3 confirm that UreD and UreF bind near UreB at the periphery of the (UreAC)3 structure. This study supports an activation model in which a domain-shifted UreB conformation in (UreABC-UreDF)3 allows CO2 and nickel ions to gain access to the nascent active site. PMID:18823937

  9. The Structure of Urease Activiation Complexes Examined by Flexibility Analysis, Mutagenesis, and Small-angle X-ray Scattering Approaches

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Quiroz, Soledad; Sukuru, Sai Chetan K.; Hausinger, Robert P.

    2008-01-01

    Conformational changes of Klebsiella aerogenes urease apoprotein (UreABC){sub 3} induced upon binding of the UreD and UreF accessory proteins were examined by a combination of flexibility analysis, mutagenesis, and small-angle X-ray scattering (SAXS). ProFlex analysis of urease provided evidence that the major domain of UreB can move in a hinge-like motion to account for prior chemical cross-linking results. Rigidification of the UreB hinge region, accomplished through a G11P mutation, reduced the extent of urease activation, in part by decreasing the nickel content of the mutant enzyme, and by sequestering a portion of the urease apoprotein in a novel activation complexmore » that includes all of the accessory proteins. SAXS analyses of urease, (UreABC-UreD){sub 3}, and (UreABC-UreDF){sub 3} confirm that UreD and UreF bind near UreB at the periphery of the (UreAC){sub 3} structure. This study supports an activation model in which a domain-shifted UreB conformation in (UreABC-UreDF){sub 3} allows CO{sub 2} and nickel ions to gain access to the nascent active site.« less

  10. Field study of vaccination of cattle with Brucella abortus strains RB51 and 19 under high and low disease prevalence.

    PubMed

    Lord, V R; Schurig, G G; Cherwonogrodzky, J W; Marcano, M J; Melendez, G E

    1998-08-01

    To assess humoral and protective immunity in cattle vaccinated by 12 months with Brucella abortus vaccine strains RB51 and 19 under field conditions of high and low brucellosis prevalence. 450 seronegative female cattle: 330 three to eight months old (calves), and 120 ten to twelve months old (heifers). Ranch A had high prevalence (39%) of brucellosis, and ranch B had low prevalence (2%), as determined by results of conventional serologic testing: agar gel immunodiffusion and the ring test. Seronegative cattle were vaccinated once or twice with 5 x 10(9) colony-forming units of B abortus strain RB51 or once with strain 19. After vaccinating 285 cattle with strain RB51 and 165 with strain 19, 74 (26%) and 30 (18%), respectively, were bred to seropositive bulls, then were kept within the infected herd of origin. All cattle vaccinated with strain 19 seroconverted 30 days later. All 285 cattle vaccinated with strain RB51 had negative results for all serologic tests, including agar gel immunodiffusion. All RB51-vaccinated cattle that became pregnant had negative results for the ring test and for conventional serologic tests after their first calving. Strain RB51 can be used as a live organism vaccine without inducing antibody titers that interfere with serodiagnosis, and induced 100% protection against field strain B abortus-induced abortion in cattle vaccinated at least 1 year before mating to an infected bull. Vaccination with strain 19 under similar conditions was less effective than vaccination with strain RB51.

  11. Infection of cattle with Brucella abortus biovar 1 isolated from a bison in Wood Buffalo National Park.

    PubMed Central

    Forbes, L B; Tessaro, S V

    1996-01-01

    An experiment was conducted to determine if cattle could be infected with a strain of Brucella abortus biovar 1 isolated from a bison in Wood Buffalo National Park. Three pregnant cows inoculated conjunctivally with 5.7 x 10(8) cfu of the bacterium, and their subsequent calves, showed seroconversion on standard serological tests for bovine brucellosis, and large numbers of the bacterium were isolated from numerous tissues at necropsy. A 4th cow that was moved into the pen that previously contained the inoculated cows subsequently showed seroconversion, and the same strain of B. abortus biovar 1 was isolated from numerous tissues. Although this strain from bison in Wood Buffalo National Park has existed in isolation from cattle for over 60 years, it remains infectious and contagious for cattle. PMID:8809394

  12. Obstetric outcomes of patients with abortus imminens in the first trimester.

    PubMed

    Evrenos, Ayşe Nur; Cakir Gungor, Ayse Nur; Gulerman, Cavidan; Cosar, Emine

    2014-03-01

    We aimed to find out the effect of abortus imminens (AI) on obstetric outcomes of pregnancies which continued beyond the 24th week of gestation. In this prospective study, 309 patients with AI were divided into high-risk group (with a risk factor for spontaneous abortus) (n = 92) and low-risk group (without a risk factor) (n = 217). The control group (n = 308) was chosen randomly. In AI group, preterm delivery, preterm premature rupture of membranes (PPROM), cesarean section (C/S) delivery, postpartum uterine atony and need of a neonatal intensive care unit (NICU) rates were significantly higher than control group. Gestational diabetes mellitus, PPROM, still birth, low APGAR scores were seen more frequently in the high-risk patients than in the control group. Furthermore in the high-risk group, preterm delivery, malpresentation, C/S delivery and need of NICU were increased much more than in the low-risk group. Gestational hypertension/preeclampsia, oligo/polyhydramniosis, intrauterine growth retardation, placenta previa, abruption of placenta, chorioamnionitis, congenital abnormalities, delivery induction, cephalopelvic disproportion, fetal distress and manual removal of placenta were not different among the groups. Patients with AI history, especially with high-risk factors can have adverse obstetric and neonatal results. So their antenatal follow-up has to be done cautiously for the early signs and symptoms of these complications.

  13. Azelainsäure 20 % Creme: Auswirkung auf Lebensqualität und Krankheitsaktivität bei erwachsenen Patientinnen mit Acne vulgaris.

    PubMed

    Kainz, Julius Thomas; Berghammer, Gabriele; Auer-Grumbach, Piet; Lackner, Verena; Perl-Convalexius, Sylvia; Popa, Rodica; Wolfesberger, Barbara

    2016-12-01

    Zur Wirksamkeit von Aknetherapien und deren Auswirkungen auf die Lebensqualität erwachsener Patienten liegen kaum Daten vor. ZIEL: Erhebung der Wirkung von Azelainsäure 20 % Creme (Skinoren ® ) auf Akne-Schweregrad und krankheitsbedingte Lebensqualität. Nichtinterventionelle Studie bei erwachsenen Patientinnen mit leichter bis mittelschwerer Akne. Wirksamkeitsparameter waren DLQI sowie Akne-Schweregrad im Gesicht, am Dekolleté sowie am Rücken im Gesamturteil des Prüfarztes (IGA-Skala: Grad 1 = annähernd reine Haut; 2 = leichte Akne; 3 = mittelschwere Akne). Visiten waren zu Studienbeginn sowie nach 4-8 und zwölf Wochen geplant. Von den 251 eingeschlossenen Patientinnen lag zu Studienbeginn bei 59 %, 31 % bzw. 10 % ein IGA-Grad von 1, 2 bzw. 3 vor; die am häufigsten betroffene Hautpartie war das Gesicht (IGA-Grad 2 oder 3: 79 %). Nach zwölf Behandlungswochen war eine signifikante Besserung der Acne vulgaris im Gesicht (IGA-Grad 0 oder 1: 82 %) sowie auf Dekolleté und Rücken feststellbar. Der mediane DLQI-Wert sank von neun zu Studienbeginn auf fünf nach zwölf Behandlungswochen. Neunzig Prozent der behandelnden Ärzte und Patientinnen beurteilten die Verträglichkeit der Behandlung als sehr gut oder gut. Die Anwendung von 20%iger Azelainsäure-Creme führt bei erwachsenen Frauen zu einer signifikanten Besserung der Acne vulgaris und der krankheitsbedingten Lebensqualität. © 2016 Deutsche Dermatologische Gesellschaft (DDG). Published by John Wiley & Sons Ltd.

  14. Detection of Brucella abortus DNA in aborted goats and sheep in Egypt by real-time PCR.

    PubMed

    Wareth, Gamal; Melzer, Falk; Tomaso, Herbert; Roesler, Uwe; Neubauer, Heinrich

    2015-06-03

    Brucellosis is a major zoonoses affects wide range of domesticated as well as wild animals. Despite the eradication program of brucellosis in Egypt, the disease is still endemic among cattle, buffaloes, sheep, goats, and camels. In the present study, abortion occurred naturally among 25 animals (10 cows, 5 buffaloes, 9 Egyptian Baladi goats and 1 ewe) shared the same pasture were investigated by real-time polymerase chain reaction (RT-PCR). DNA of Brucella (B.) abortus was detected in serum of goats and sheep which has aborted recently by species-specific RT-PCR. The results suggest cross-species infection of B. abortus from cattle to non-preferred hosts raised in close contact. This article will renew our knowledge about the Brucella agent causing abortion in small ruminants in Egypt. Information provided in this study is important for surveillance program, because eradication programs and vaccination strategies may have to be adapted accordingly.

  15. TLR2 and TLR4 signaling pathways are required for recombinant Brucella abortus BCSP31-induced cytokine production, functional upregulation of mouse macrophages, and the Th1 immune response in vivo and in vitro

    PubMed Central

    Li, Jia-Yun; Liu, Yuan; Gao, Xiao-Xue; Gao, Xiang; Cai, Hong

    2014-01-01

    Brucella abortus is a zoonotic Gram-negative pathogen that causes brucelosis in ruminants and humans. Toll-like receptors (TLRs) recognize Brucella abortus and initiate antigen-presenting cell activities that affect both innate and adaptive immunity. In this study, we focused on recombinant Brucella cell-surface protein 31 (rBCSP31) to determine its effects on mouse macrophages. Our results demonstrated that rBCSP31 induced TNF-α, IL-6 and IL-12p40 production, which depended on the activation of mitogen-activated protein kinases (MAPKs) by stimulating the rapid phosphorylation of p38 and JNK and the activation of transcription factor NF-κB in macrophages. In addition, continuous exposure (>24 h) of RAW264.7 cells to rBCSP31 significantly enhanced IFN-γ-induced expression of MHC-II and the ability to present rBCSP31 peptide to CD4+ T cells. Furthermore, we found that rBCSP31 could interact with both TLR2 and TLR4. The rBCSP31-induced cytokine production by macrophages from TLR2−/− and TLR4−/− mice was lower than that from C57BL/6 macrophages, and the activation of NF-κB and MAPKs was attenuated in macrophages from TLR2−/− and TLR4−/− mice. In addition, CD4+ T cells from C57BL/6 mice immunized with rBCSP31 produced higher levels of IFN-γ and IL-2 compared with CD4+ T cells from TLR2−/− and TLR4−/− mice. Macrophages from immunized C57BL/6 mice produced higher levels of IL-12p40 than those from TLR2−/− and TLR4−/− mice. Furthermore, immunization with rBCSP31 provided better protection in C57BL/6 mice than in TLR2−/− and TLR4−/− mice after B. abortus 2308 challenge. These results indicate that rBCSP31 is a TLR2 and TLR4 agonist that induces cytokine production, upregulates macrophage function and induces the Th1 immune response. PMID:24769793

  16. Molecular Epidemiology of Brucella abortus Isolates from Cattle, Elk, and Bison in the United States, 1998 to 2011

    PubMed Central

    Stuber, Tod; Quance, Christine; Edwards, William H.; Tiller, Rebekah V.; Linfield, Tom; Rhyan, Jack; Berte, Angela; Harris, Beth

    2012-01-01

    A variable-number tandem repeat (VNTR) protocol targeting 10 loci in the Brucella abortus genome was used to assess genetic diversity among 366 field isolates recovered from cattle, bison, and elk in the Greater Yellowstone Area (GYA) and Texas during 1998 to 2011. Minimum spanning tree (MST) and unweighted-pair group method with arithmetic mean (UPGMA) analyses of VNTR data identified 237 different VNTR types, among which 14 prominent clusters of isolates could be identified. Cattle isolates from Texas segregated into three clusters: one comprised of field isolates from 1998 to 2005, one comprised of vaccination-associated infections, and one associated with an outbreak in Starr County in January 2011. An isolate obtained from a feral sow trapped on property adjacent to the Starr County herd in May 2011 clustered with the cattle isolates, suggesting a role for feral swine as B. abortus reservoirs in Starr County. Isolates from a 2005 cattle outbreak in Wyoming displayed VNTR-10 profiles matching those of strains recovered from Wyoming and Idaho elk. Additionally, isolates associated with cattle outbreaks in Idaho in 2002, Montana in 2008 and 2011, and Wyoming in 2010 primarily clustered with isolates recovered from GYA elk. This study indicates that elk play a predominant role in the transmission of B. abortus to cattle located in the GYA. PMID:22427502

  17. Vector Development for the Expression of Foreign Proteins in the Vaccine Strain Brucella abortus S19

    PubMed Central

    Comerci, Diego J.; Pollevick, Guido D.; Vigliocco, Ana M.; Frasch, Alberto C. C.; Ugalde, Rodolfo A.

    1998-01-01

    A vector for the expression of foreign antigens in the vaccine strain Brucella abortus S19 was developed by using a DNA fragment containing the regulatory sequences and the signal peptide of the Brucella bcsp31 gene. This fragment was cloned in broad-host-range plasmid pBBR4MCS, resulting in plasmid pBEV. As a reporter protein, a repetitive antigen of Trypanosoma cruzi was used. The recombinant fusion protein is stably expressed and secreted into the Brucella periplasmic space, inducing a good antibody response against the T. cruzi antigen. The expression of the repetitive antigen in Brucella neither altered its growth pattern nor generated a toxic or lethal effect during experimental infection. The application of this strategy for the generation of live recombinant vaccines and the tagging of B. abortus S19 vaccine is discussed. This is the first time that a recombinant protein has been expressed in the periplasm of brucellae. PMID:9673273

  18. Differential composition of culture supernatants from wild-type Brucella abortus and its isogenic virB mutants.

    PubMed

    Delpino, M Victoria; Comerci, Diego J; Wagner, Mary Ann; Eschenbrenner, Michel; Mujer, Cesar V; Ugalde, Rodolfo A; Fossati, Carlos A; Baldi, Pablo C; Delvecchio, Vito G

    2009-07-01

    The virB genes coding type IV secretion system are necessary for the intracellular survival and replication of Brucella spp. In this study, extracellular proteins from B. abortus 2308 (wild type, WT) and its isogenic virB10 polar mutant were compared. Culture supernatants harvested in the early stationary phase were concentrated and subjected to 2D electrophoresis. Spots present in the WT strain but absent in the virB10 mutant (differential spots) were considered extracellular proteins released in a virB-related manner, and were identified by MALDI-TOF analysis and matching with Brucella genomes. Among the 11 differential proteins identified, DnaK chaperone (Hsp70), choloylglycine hydrolase (CGH) and a peptidyl-prolyl cis-trans isomerase (PPIase) were chosen for further investigation because of their homology with extracellular and/or virulence factors from other bacteria. The three proteins were obtained in recombinant form and specific monoclonal antibodies (mAbs) were prepared. By Western blot with these mAbs, the three proteins were detected in supernatants from the WT but not in those from the virB10 polar mutant or from strains carrying non-polar mutations in virB10 or virB11 genes. These results suggest that the expression of virB genes affects the extracellular release of DnaK, PPIase and CGH, and possibly other proteins from B. abortus.

  19. Immune responses of bison and efficacy after booster vaccination with Brucella abortus strain RB51

    USDA-ARS?s Scientific Manuscript database

    Thirty-one bison heifers were randomly assigned to saline (control; n=7) or single vaccination (n=24) with 1010 CFU of B. abortus strain RB51 (RB51). Some vaccinated bison were randomly selected for booster vaccination with 10**10 CFU of RB51 at 11 months after initial vaccination (n=16). When comp...

  20. Efficacy of single calfhood vaccination of elk with Brucella abortus strain 19

    USGS Publications Warehouse

    Roffe, T.J.; Jones, L.C.; Coffin, K.; Drew, M.L.; Sweeney, Steven J.; Hagius, S.D.; Elzer, P.H.; Davis, D.

    2004-01-01

    Brucellosis has been eradicated from cattle in the states of Wyoming, Montana, and Idaho, USA. However, free-ranging elk (Cervus elaphus) that use feedgrounds in the Greater Yellowstone Area (GYA) and bison (Bison bison) in Yellowstone and Grand Teton national parks still have high seroprevalence to the disease and have caused loss of brucellosis-free status in Wyoming. Management tools to control or eliminate the disease are limited; however, wildlife vaccination is among the methods currently used by wildlife managers in Wyoming. We conducted a controlled challenge study of single calfhood vaccination. Elk calves, caught in January and February of 1999 and 2000 and acclimated to captivity for 3 weeks, were randomly assigned to control or vaccinate groups. The vaccinate groups received Brucetta abortus vaccine strain 19 (S19) by hand-delivered intramuscular injection. Calves were raised to adulthood and bred at either 2.5 or 3.5 years of age for 2000 and 1999 captures, respectively. Eighty-nine (44 controls, 45 vaccinates) pregnant elk entered the challenge portion of the study. We challenged elk at mid-gestation with pathogenic B. abortus strain 2308 by intraconjunctival instillation. Abortion occurred in significantly more (P = 0.002) controls (42; 93%) than vaccinates (32; 71%), and vaccine protected 25% of the vaccinate group. We used Brucella culture of fetus/calf tissues to determine the efficacy of vaccination for preventing infection, and we found that the number of infected fetuses/calves did not differ between controls and vaccinates (P = 0.14). Based on these data, single calfhood vaccination with S19 has low efficacy, will likely have only little to moderate effect on Brucella prevalence in elk, and is unlikely to eradicate the disease in wildlife of the GYA.

  1. Inhibitory Effect of the Ethanol Extract of a Rice Bran Mixture Comprising Angelica gigas, Cnidium officinale, Artemisia princeps, and Camellia sinensis on Brucella abortus Uptake by Professional and Nonprofessional Phagocytes.

    PubMed

    Hop, Huynh Tan; Arayan, Lauren Togonon; Reyes, Alisha Wehdnesday Bernardo; Huy, Tran Xuan Ngoc; Baek, Eun Jin; Min, WonGi; Lee, Hu Jang; Lee, Chun Hee; Rhee, Man Hee; Kim, Suk

    2017-10-28

    In this study, we evaluated the inhibitory effect of a rice bran mixture extract (RBE) on Brucella abortus pathogenesis in professional (RAW 264.7) and nonprofessional (HeLa) phagocytes. We fermented the rice bran mixture and then extracted it with 50% ethanol followed by gas chromatography-mass spectrometry to identify the components in RBE. Our results clearly showed that RBE caused a significant reduction in the adherence of B. abortus in both cell lines. Furthermore, analysis of phagocytic signaling proteins by western blot assay revealed that RBE pretreatment resulted in inhibition of phosphorylation of JNK, ERK, and p38, leading to decline of internalization compared with the controls. Additionally, the intensity of F-actin observed by fluorescence microscopy and FACS was remarkably reduced in RBE-pretreated cells compared with control cells. However, the intracellular replication of B. abortus within phagocytes was not affected by RBE. Taken together, these findings suggest that the phagocytic receptor blocking and suppressive effects of RBE on the MAPK-linked phagocytic signaling pathway could negatively affect the invasion of B. abortus into phagocytes.

  2. Evaluation of DNA extraction protocols for Brucella abortus pcr detection in aborted fetuses or calves born from cows experimentally infected with strain 2308

    PubMed Central

    Matrone, M.; Keid, L.B.; Rocha, V.C.M.; Vejarano, M.P.; Ikuta, C.Y.; Rodriguez, C.A.R.; Ferreira, F.; Dias, R.A.; Ferreira Neto, J.S

    2009-01-01

    The objective of the present study was to improve the detection of B. abortus by PCR in organs of aborted fetuses from infected cows, an important mechanism to find infected herds on the eradication phase of the program. So, different DNA extraction protocols were compared, focusing the PCR detection of B. abortus in clinical samples collected from aborted fetuses or calves born from cows challenged with the 2308 B. abortus strain. Therefore, two gold standard groups were built based on classical bacteriology, formed from: 32 lungs (17 positives), 26 spleens (11 positives), 23 livers (8 positives) and 22 bronchial lymph nodes (7 positives). All samples were submitted to three DNA extraction protocols, followed by the same amplification process with the primers B4 and B5. From the accumulated results for organ, the proportion of positives for the lungs was higher than the livers (p=0.04) or bronchial lymph nodes (p=0.004) and equal to the spleens (p=0.18). From the accumulated results for DNA extraction protocol, the proportion of positives for the Boom protocol was bigger than the PK (p< 0.0001) and GT (p=0.0004). There was no difference between the PK and GT protocols (p=0.5). Some positive samples from the classical bacteriology were negative to the PCR and vice-versa. Therefore, the best strategy for B. abortus detection in the organs of aborted fetuses or calves born from infected cows is the use, in parallel, of isolation by classical bacteriology and the PCR, with the DNA extraction performed by the Boom protocol. PMID:24031391

  3. A Live Vaccine from Brucella abortus Strain 82 for Control of Cattle Brucellosis in the Russian Federation

    USDA-ARS?s Scientific Manuscript database

    During the first half of the 20th century, widespread regulatory efforts to control cattle brucellosis (Brucella abortus) in the Union of Soviet Socialist Republics were essentially nonexistent, and control was limited to selective test and slaughter of serologic agglutination reactors. By the 1950...

  4. Identification and characterization of the nickel uptake system for urease biogenesis in Streptococcus salivarius 57.I.

    PubMed

    Chen, Yi-Ywan M; Burne, Robert A

    2003-12-01

    Ureases are multisubunit enzymes requiring Ni(2+) for activity. The low pH-inducible urease gene cluster in Streptococcus salivarius 57.I is organized as an operon, beginning with ureI, followed by ureABC (structural genes), and ureEFGD (accessory genes). Urease biogenesis also requires a high-affinity Ni(2+) uptake system. By searching the partial genome sequence of a closely related organism, Streptococcus thermophilus LMG18311, three open reading frame (ORFs) homologous to those encoding proteins involved in cobalamin biosynthesis and cobalt transport (cbiMQO) were identified immediately 3' to the ure operon. To determine whether these genes were involved in urease biogenesis by catalyzing Ni(2+) uptake in S. salivarius, regions 3' to ureD were amplified by PCRs from S. salivarius by using primers identical to the S. thermophilus sequences. Sequence analysis of the products revealed three ORFs. Reverse transcriptase PCR was used to demonstrate that the ORFs are transcribed as part of the ure operon. Insertional inactivation of ORF1 with a polar kanamycin marker completely abolished urease activity and the ability to accumulate (63)Ni(2+) during growth. Supplementation of the growth medium with NiCl(2) at concentrations as low as 2.5 micro M partially restored urease activity in the mutant. Both wild-type and mutant strains showed enhanced urease activity when exogenous Ni(2+) was provided at neutral pH. Enhancement of urease activity by adding nickel was regulated at the posttranslational level. Thus, ORF1, ORF2, and ORF3 are part of the ure operon, and these genes, designated ureM, ureQ, and ureO, respectively, likely encode a Ni(2+)-specific ATP-binding cassette transporter.

  5. The bhuQ Gene Encodes a Heme Oxygenase That Contributes to the Ability of Brucella abortus 2308 To Use Heme as an Iron Source and Is Regulated by Irr

    PubMed Central

    Ojeda, Jenifer F.; Martinson, David A.; Menscher, Evan A.

    2012-01-01

    The Brucella BhuQ protein is a homolog of the Bradyrhizobium japonicum heme oxygenases HmuD and HmuQ. To determine if this protein plays a role in the ability of Brucella abortus 2308 to use heme as an iron source, an isogenic bhuQ mutant was constructed and its phenotype evaluated. Although the Brucella abortus bhuQ mutant DCO1 did not exhibit a defect in its capacity to use heme as an iron source or evidence of increased heme toxicity in vitro, this mutant produced increased levels of siderophore in response to iron deprivation compared to 2308. Introduction of a bhuQ mutation into the B. abortus dhbC mutant BHB2 (which cannot produce siderophores) resulted in a severe growth defect in the dhbC bhuQ double mutant JFO1 during cultivation under iron-restricted conditions, which could be rescued by the addition of FeCl3, but not heme, to the growth medium. The bhuQ gene is cotranscribed with the gene encoding the iron-responsive regulator RirA, and both of these genes are repressed by the other major iron-responsive regulator in the alphaproteobacteria, Irr. The results of these studies suggest that B. abortus 2308 has at least one other heme oxygenase that works in concert with BhuQ to allow this strain to efficiently use heme as an iron source. The genetic organization of the rirA-bhuQ operon also provides the basis for the proposition that BhuQ may perform a previously unrecognized function by allowing the transcriptional regulator RirA to recognize heme as an iron source. PMID:22636783

  6. Structural, functional and immunogenic insights on Cu,Zn Superoxide Dismutase pathogenic virulence factors from Neisseria meningitidis and Brucella abortus

    USDA-ARS?s Scientific Manuscript database

    Bacterial pathogens Neisseria meningitidis and Brucella abortus pose threats to human and animal health worldwide, causing meningococcal disease and brucellosis, respectively. Mortality from acute N. meningitidis infections remains high despite antibiotics, and brucellosis presents alimentary and he...

  7. An ecological perspective on the changing face of Brucella abortus in the western United States

    USGS Publications Warehouse

    Cross, Paul C.; Maichak, Eric J.; Brennan, Angela; Scurlock, Brandon M.; Henningsen, John C.; Luikart, Gordon

    2013-01-01

    After a hiatus during the 1990s, outbreaks of Brucella abortus in cattle are occurring more frequently in some of the western states of the United States, namely, Montana, Wyoming and Idaho. This increase is coincident with increasing brucellosis seroprevalence in elk (Cervus elaphus), which is correlated with elk density. Vaccines are a seductive solution, but their use in wildlife systems remains limited by logistical, financial, and scientific constraints. Cattle vaccination is ongoing in the region. Livestock regulations, however, tend to be based on serological tests that test for previous exposure and available vaccines do not protect against seroconversion. The authors review recent ecological studies of brucellosis, with particular emphasis on the Greater Yellowstone Area, and highlight the management options and implications of this work, including the potential utility of habitat modifications and targeted hunts, as well as scavengers and predators. Finally, the authors discuss future research directions that will help us to understand and manage brucellosis in wildlife.

  8. Early transcriptional responses of internalization defective Brucella abortus mutants in professional phagocytes, RAW 264.7.

    PubMed

    Cha, Seung Bin; Lee, Won Jung; Shin, Min Kyoung; Jung, Myung Hwan; Shin, Seung Won; Yoo, An Na; Kim, Jong Wan; Yoo, Han Sang

    2013-06-27

    Brucella abortus is an intracellular zoonotic pathogen which causes undulant fever, endocarditis, arthritis and osteomyelitis in human and abortion and infertility in cattle. This bacterium is able to invade and replicate in host macrophage instead of getting removed by this defense mechanism. Therefore, understanding the interaction between virulence of the bacteria and the host cell is important to control brucellosis. Previously, we generated internalization defective mutants and analyzed the envelope proteins. The present study was undertaken to evaluate the changes in early transcriptional responses between wild type and internalization defective mutants infected mouse macrophage, RAW 264.7. Both of the wild type and mutant infected macrophages showed increased expression levels in proinflammatory cytokines, chemokines, apoptosis and G-protein coupled receptors (Gpr84, Gpr109a and Adora2b) while the genes related with small GTPase which mediate intracellular trafficking was decreased. Moreover, cytohesin 1 interacting protein (Cytip) and genes related to ubiquitination (Arrdc3 and Fbxo21) were down-regulated, suggesting the survival strategy of this bacterium. However, we could not detect any significant changes in the mutant infected groups compared to the wild type infected group. In summary, it was very difficult to clarify the alterations in host cellular transcription in response to infection with internalization defective mutants. However, we found several novel gene changes related to the GPCR system, ubiquitin-proteosome system, and growth arrest and DNA damages in response to B. abortus infection. These findings may contribute to a better understanding of the molecular mechanisms underlying host-pathogen interactions and need to be studied further.

  9. Comparison of abortion and infection after experimental challenge of pregnant bison and cattle with Brucella abortus strain 2308

    USDA-ARS?s Scientific Manuscript database

    A comparative study was conducted using data from naive bison (n=45) and cattle (n=46) from 8 and 6 studies, respectively, in which a standardized Brucella abortus strain 2308 experimental challenge was administered. The incidence of abortion, fetal infection, uterine or mammary infection, or infec...

  10. Efficacy of dart or booster vaccination with strain RB51 in protecting bison against experimental Brucella abortus challenge

    USDA-ARS?s Scientific Manuscript database

    Vaccination is an effective tool for reducing the prevalence of brucellosis in natural hosts. In this study, we characterized the efficacy of the Brucella abortus strain RB51 (RB51) vaccine in bison when delivered by single intramuscular vaccination (Hand RB51), single pneumatic dart delivery (Dart ...

  11. The Bactericidal Effect of High Temperature Is an Essential Resistance Mechanism of Chicken Macrophage against Brucella abortus Infection.

    PubMed

    Arayan, Lauren Togonon; Reyes, Alisha Wehdnesday Bernardo; Hop, Huynh Tan; Xuan, Huy Tran; Baek, Eun Jin; Min, Wongi; Kim, Suk

    2017-10-28

    Knowledge of avian host responses to brucellosis is critical to understanding how birds resist this infection; however, this mechanism is not well established. On the other hand, temperature has a major involvement in the physiology of living organisms, and cell death induced by heat is attributed to protein denaturation. This study demonstrates the direct bactericidal effect of a high temperature (41ºC) on Brucella abortus that resulted in the gradual reduction of intracellular bacteria and inhibited bacterial growth within avian macrophage HD11 in an increasing period of time. On the other hand, this study also revealed that high temperature does not affect the rate of bacterial uptake, as confirmed by the bacterial adherence assay. No significant difference was observed in the expression of target genes between infected and uninfected cells for both temperatures. This study suggests the susceptibility of B. abortus to bacterial death under a high temperature with an increased period of incubation, leading to suppression of bacterial growth.

  12. The relationship of initial embryo crown--rump length to pregnancy outcome and abortus karyotype based on new growth curves for the 2-31 mm embryo.

    PubMed

    Dickey, R P; Gasser, R F; Olar, T T; Curole, D N; Taylor, S N; Matulich, E M; West, J D; Tsien, F

    1994-02-01

    The objective of this study was to determine if measurement of initial crown--rump length (CRL) is helpful in predicting low birth weight, newborn length, spontaneous abortions, or abortus karyotype. We measured CRL prospectively in 837 consecutive singleton pregnancies at the time a heart rate was first detectable with transvaginal ultrasonography and compared these measurements to normal values for the 10th through 90th centiles determined from 227 transvaginal ultrasound measurements in in-vitro fertilization and gamete intra-Fallopian transfer pregnancies with known ovulation dates. The relationship of initial CRL to birth weight and length and to abortion and abortus karyotype was analysed after all pregnancies had delivered. Initial CRL measured after the 28th post-ovulation day was predictive of subsequent abortion, but not of low birth weight or length. The abortion rate was 3.3% [95% confidence interval (CI) 1.5%, 5.1%] when initial CRL > or = 50th centile, compared to 19.4% (95% CI 15.4%, 23.4%) when < 50th centile. Initial CRL was < 50th centile in 13 out of 14 trisomic and in eight out of 10 other karyotypically abnormal aborti. These results indicate that initial CRL measured after the 28th post-ovulation day may help to identify pregnancies at increased risk of abortion due to abnormal karyotypes.

  13. Early transcriptional responses of internalization defective Brucella abortus mutants in professional phagocytes, RAW 264.7

    PubMed Central

    2013-01-01

    Background Brucella abortus is an intracellular zoonotic pathogen which causes undulant fever, endocarditis, arthritis and osteomyelitis in human and abortion and infertility in cattle. This bacterium is able to invade and replicate in host macrophage instead of getting removed by this defense mechanism. Therefore, understanding the interaction between virulence of the bacteria and the host cell is important to control brucellosis. Previously, we generated internalization defective mutants and analyzed the envelope proteins. The present study was undertaken to evaluate the changes in early transcriptional responses between wild type and internalization defective mutants infected mouse macrophage, RAW 264.7. Results Both of the wild type and mutant infected macrophages showed increased expression levels in proinflammatory cytokines, chemokines, apoptosis and G-protein coupled receptors (Gpr84, Gpr109a and Adora2b) while the genes related with small GTPase which mediate intracellular trafficking was decreased. Moreover, cytohesin 1 interacting protein (Cytip) and genes related to ubiquitination (Arrdc3 and Fbxo21) were down-regulated, suggesting the survival strategy of this bacterium. However, we could not detect any significant changes in the mutant infected groups compared to the wild type infected group. Conclusions In summary, it was very difficult to clarify the alterations in host cellular transcription in response to infection with internalization defective mutants. However, we found several novel gene changes related to the GPCR system, ubiquitin-proteosome system, and growth arrest and DNA damages in response to B. abortus infection. These findings may contribute to a better understanding of the molecular mechanisms underlying host-pathogen interactions and need to be studied further. PMID:23802650

  14. Comparison of Abortion and Infection after Experimental Challenge of Pregnant Bison and Cattle with Brucella abortus Strain 2308▿

    PubMed Central

    Olsen, S. C.; Johnson, C.

    2011-01-01

    A comparative study was conducted using data from naive bison (n = 45) and cattle (n = 46) from 8 and 6 studies, respectively, in which a standardized Brucella abortus strain 2308 experimental challenge was administered during midgestation. The incidence of abortion, fetal infection, uterine or mammary infection, or infection in maternal tissues after experimental challenge was greater (P < 0.05) in bison than in cattle. In animals that did abort, the time between experimental challenge and abortion was shorter (P < 0.05) for bison than for cattle. Brucella colonization of four target tissues and serologic responses on the standard tube agglutination test at the time of abortion did not differ (P > 0.05) between cattle and bison. The results of our study suggest that naive bison and cattle have similarities and differences after experimental exposure to a virulent B. abortus strain. Although our data suggest that bison may be more susceptible to infection with Brucella, some pathogenic characteristics of brucellosis were similar between bison and cattle. PMID:21976222

  15. Interaction Network and Localization of Brucella abortus Membrane Proteins Involved in the Synthesis, Transport, and Succinylation of Cyclic β-1,2-Glucans

    PubMed Central

    Guidolin, Leticia S.; Morrone Seijo, Susana M.; Guaimas, Francisco F.

    2015-01-01

    ABSTRACT Cyclic β-1,2-glucans (CβG) are periplasmic homopolysaccharides that play an important role in the virulence and interaction of Brucella with the host. Once synthesized in the cytoplasm by the CβG synthase (Cgs), CβG are transported to the periplasm by the CβG transporter (Cgt) and succinylated by the CβG modifier enzyme (Cgm). Here, we used a bacterial two-hybrid system and coimmunoprecipitation techniques to study the interaction network between these three integral inner membrane proteins. Our results indicate that Cgs, Cgt, and Cgm can form both homotypic and heterotypic interactions. Analyses carried out with Cgs mutants revealed that the N-terminal region of the protein (Cgs region 1 to 418) is required to sustain the interactions with Cgt and Cgm as well as with itself. We demonstrated by single-cell fluorescence analysis that in Brucella, Cgs and Cgt are focally distributed in the membrane, particularly at the cell poles, whereas Cgm is mostly distributed throughout the membrane with a slight accumulation at the poles colocalizing with the other partners. In summary, our results demonstrate that Cgs, Cgt, and Cgm form a membrane-associated biosynthetic complex. We propose that the formation of a membrane complex could serve as a mechanism to ensure the fidelity of CβG biosynthesis by coordinating their synthesis with the transport and modification. IMPORTANCE In this study, we analyzed the interaction and localization of the proteins involved in the synthesis, transport, and modification of Brucella abortus cyclic β-1,2-glucans (CβG), which play an important role in the virulence and interaction of Brucella with the host. We demonstrate that these proteins interact, forming a complex located mainly at the cell poles; this is the first experimental evidence of the existence of a multienzymatic complex involved in the metabolism of osmoregulated periplasmic glucans in bacteria and argues for another example of pole differentiation in Brucella

  16. Development of an in vivo model of Chlamydia abortus chronic infection in mice overexpressing IL-10.

    PubMed

    Del Río, Laura; Murcia, Antonio; Buendía, Antonio J; Álvarez, Daniel; Ortega, Nieves; Navarro, José A; Salinas, Jesús; Caro, María Rosa

    2018-01-01

    Chlamydia abortus, like other members of the family Chlamydiaceae, have a unique intracellular developmental cycle that is characterized by its chronic nature. Infection of a flock can remain undetected for months, until abortion occurs the following reproductive season but, to date, neither the location nor the mechanisms that maintain this latent phase are fully understood. Studies have shown that IL-10 produced as a response to certain micro-organisms sustains the intracellular survival of pathogens and increases host susceptibility to chlamydial infections. In order to induce a sustained infection C. abortus, transgenic mice that constitutively express IL-10 were infected and the immunological mechanisms that maintain infection in these mice were compared with the mechanisms of a resistant wild-type mouse strain. Viable bacteria could be detected in different tissues of transgenic mice up to 28 days after infection, as analysed by bacterial isolation and immunohistochemistry. Chronic infection in these mice was associated with an impaired recruitment of macrophages, decreased iNOS activity at the site of infection and a more diffuse distribution of inflammatory cells in the liver. This murine model can be of great help for understanding the immunological and bacterial mechanisms that lead to chronic chlamydial infections. Copyright © 2017 Elsevier B.V. All rights reserved.

  17. Serological profile of buffalo (Bubalus bubalis) female calves vaccinated with standard Brucella abortus strain 19 vaccine using rose bengal, 2-mercaptoethanol and complement fixation tests.

    PubMed

    Nardi, G Júnior; Ribeiro, M G; Jorge, A M; Megid, J; Silva, L M P

    2012-03-01

    The serological profiles of 21 female buffaloes vaccinated between 3 and 8 months of age using Brucella abortus strain 19 (S19) were evaluated by rose bengal (RBT), 2-mercaptoethanol (2ME) and complement fixation (CFT) tests. The serum strains were collected in day zero, 15, 30, 45, 60th days and subsequently to each 30 months, until 720th day after vaccination. No animal showed reaction in day zero. In 15th day above 95% of animals revealed reaction in all tests. All the animals presented absence of reactions in CFT, RBT and 2ME tests at 270, 300 and 360 days after vaccination, respectively. Our finding highlighted early response in CFT compared than other conventional agglutination tests. None of animals presented oscillation of titers or reactions in any test after 360 day of study, which enables the use of these tests after this period without interference of antibodies from S19 vaccine origin between 3 and 8 months in buffalo heifers. Copyright © 2011 The International Alliance for Biological Standardization. Published by Elsevier Ltd. All rights reserved.

  18. Mercaptursäure und Nukleosidaddukt im Harn als Biomarker in 1-Hydroxymethylpyren-exponierten Ratten

    NASA Astrophysics Data System (ADS)

    Ma, Lan

    2002-01-01

    1-Methylpyren (MP) ist hepatokanzerogen in neugeborenen männlichen Mäusen. Durch Hydroxylierung an der benzylischen Stelle und anschließende Sulfonierung wird MP zu DNA-reaktivem 1-Sulfooxymethylpyren (SMP) aktiviert. In der Ratte führt die Exposition des benzylischen Alkohols, 1-Hydroxymethylpyren (HMP), zur DNA-Adduktbildung in verschiedenen Geweben. Eventuelle Konsequenz der Toxifizierung ist die Ausscheidung entsprechender Mercaptursäure und Nukleosidaddukt im Harn, welche aufgrund ihrer Herkunft als Biomarker eignen könnten. In dieser Arbeit wird die Ausscheidung der Mercaptursäure und des N2-Desoxyguanosinadduktes in HMP-exponierten Ratten untersucht. Nach der Applikation von HMP bzw. MP wurden weniger als 1 % der Dosis als MPMA über Urin und Faeces ausgeschieden (0 - 48 h). Die Ausscheidung erfolgt hauptsächlich in den ersten 24 h nach der Applikation. MPdG konnte weder in Urin noch in Faeces der HMP-behandelten Tieren identifiziert werden. Nach direkter SMP-Applikation wurde MPdG nur in sehr geringe Menge (weniger als 0,9 ppm in 12 h) im Urin gefunden. Aufgrund der geringen Menge eignet sich MPdG nicht als Biomarker. MPMA dagegen, lässt sich analytisch gut erfassen. Es sollte daher untersucht werden, ob MPMA die Toxifizierung des HMP wiederspiegelt. Die Voraussetzung dafür ist die Kenntnisse über das Metabolismusmuster von HMP. Es wurde daher umfassende Untersuchungen zum Metabolismus des HMP durchgeführt. Die Ergebnisse zeigten, dass mehr als 80 % der Metaboiten in ihrer oxidierten Form (PCS, deren Glucuronsäure-Konjugate sowie phenolische Sulfatester der PCS) ausgeschieden wurden. Demnach spielt die Oxidation des HMP zu PCS eine sehr wichtige Rolle bei der Detoxifizierung und Ausscheidung von HMP. Ferne konnte nachgewiesen werden, dass die Enzyme Alkohol- und Aldehyd-Dehydrogenase an der Oxidation von HMP beteiligt waren. Die Inhibitoren Disulfiram und Ethanol der o. g. Enzyme wurde daher zur Modulation der Detoxifizierung in vivo eingesetzt

  19. The International Standard for Anti-Brucella abortus Serum

    PubMed Central

    Stableforth, A. W.

    1954-01-01

    In field trials on the eradication of brucellosis from dairy herds in Great Britain, which began in 1933, a serum standard of reference was used for the examination of agglutinating suspensions prepared in different laboratories. In 1937, the Office International des Epizooties (OIE) adopted this standard and made recommendations for its use internationally. These recommendations were revised by OIE in 1948, by the Third Inter-American Congress on Brucellosis and by the Joint FAO/WHO Expert Panel on Brucellosis in 1950, and again by the latter body in 1952. A new batch equivalent in potency to the original standard was established by the WHO Expert Committee on Biological Standardization in 1952 as the International Standard for Anti-Brucella abortus Serum. The International Standard, or a national standard of equivalent potency, ensures comparability of the titres obtained in different countries by different methods, and the results of such comparisons can be expressed in a simple manner by describing the titres in terms of International Units of Brucella antibody. PMID:13199656

  20. Global Vision Impairment and Blindness Due to Uncorrected Refractive Error, 1990-2010.

    PubMed

    Naidoo, Kovin S; Leasher, Janet; Bourne, Rupert R; Flaxman, Seth R; Jonas, Jost B; Keeffe, Jill; Limburg, Hans; Pesudovs, Konrad; Price, Holly; White, Richard A; Wong, Tien Y; Taylor, Hugh R; Resnikoff, Serge

    2016-03-01

    The purpose of this systematic review was to estimate worldwide the number of people with moderate and severe visual impairment (MSVI; presenting visual acuity <6/18, ≥3/60) or blindness (presenting visual acuity <3/60) due to uncorrected refractive error (URE), to estimate trends in prevalence from 1990 to 2010, and to analyze regional differences. The review focuses on uncorrected refractive error which is now the most common cause of avoidable visual impairment globally. : The systematic review of 14,908 relevant manuscripts from 1990 to 2010 using Medline, Embase, and WHOLIS yielded 243 high-quality, population-based cross-sectional studies which informed a meta-analysis of trends by region. The results showed that in 2010, 6.8 million (95% confidence interval [CI]: 4.7-8.8 million) people were blind (7.9% increase from 1990) and 101.2 million (95% CI: 87.88-125.5 million) vision impaired due to URE (15% increase since 1990), while the global population increased by 30% (1990-2010). The all-age age-standardized prevalence of URE blindness decreased 33% from 0.2% (95% CI: 0.1-0.2%) in 1990 to 0.1% (95% CI: 0.1-0.1%) in 2010, whereas the prevalence of URE MSVI decreased 25% from 2.1% (95% CI: 1.6-2.4%) in 1990 to 1.5% (95% CI: 1.3-1.9%) in 2010. In 2010, URE contributed 20.9% (95% CI: 15.2-25.9%) of all blindness and 52.9% (95% CI: 47.2-57.3%) of all MSVI worldwide. The contribution of URE to all MSVI ranged from 44.2 to 48.1% in all regions except in South Asia which was at 65.4% (95% CI: 62-72%). : We conclude that in 2010, uncorrected refractive error continues as the leading cause of vision impairment and the second leading cause of blindness worldwide, affecting a total of 108 million people or 1 in 90 persons.

  1. Immunological response to Brucella abortus strain 19 vaccination of cattle in a communal area in South Africa.

    PubMed

    Simpson, Gregory J G; Marcotty, Tanguy; Rouille, Elodie; Chilundo, Abel; Letteson, Jean-Jacques; Godfroid, Jacques

    2018-03-29

    Brucellosis is of worldwide economic and public health importance. Heifer vaccination with live attenuated Brucella abortus strain 19 (S19) is the cornerstone of control in low- and middle-income countries. Antibody persistence induced by S19 is directly correlated with the number of colony-forming units (CFU) per dose. There are two vaccination methods: a 'high' dose (5-8 × 1010 CFU) subcutaneously injected or one or two 'low' doses (5 × 109 CFU) through the conjunctival route. This study aimed to evaluate serological reactions to the 'high' dose and possible implications of the serological findings on disease control. This study included 58 female cases, vaccinated at Day 0, and 29 male controls. Serum was drawn repeatedly and tested for Brucella antibodies using the Rose Bengal Test (RBT) and an indirect enzyme-linked immunosorbent assay (iELISA). The cases showed a rapid antibody response with peak RBT positivity (98%) at 2 weeks and iELISA (95%) at 8 weeks, then decreased in an inverse logistic curve to 14% RBT and 32% iELISA positive at 59 weeks and at 4.5 years 57% (4/7 cases) demonstrated a persistent immune response (RBT, iELISA or Brucellin skin test) to Brucella spp. Our study is the first of its kind documenting the persistence of antibodies in an African communal farming setting for over a year to years after 'high' dose S19 vaccination, which can be difficult to differentiate from a response to infection with wild-type B. abortus. A recommendation could be using a 'low' dose or different route of vaccination.

  2. Pseudorabies Virus and Brucella abortus from an Expanding Wild Pig ( Sus scrofa ) Population in Southern Oklahoma, USA.

    PubMed

    Gaskamp, Joshua A; Gee, Kenneth L; Campbell, Tyler A; Silvy, Nova J; Webb, Stephen L

    2016-04-28

    Wild pigs ( Sus scrofa ) are causing increasing ecologic and economic damage at a global scale. Because wild pigs can carry ≥65 diseases that affect livestock, their widespread expansion threatens native wildlife and livestock. We screened wild pigs from south-central Oklahoma, US for antibodies against Brucella abortus , pseudorabies virus (PRV), and porcine reproductive and respiratory syndrome virus (PRRS). These pathogens were chosen because they are part of eradication programs in the US and could have large economic impacts on domestic livestock if transmitted from wild animals. We tested 282 serum samples during spring 2010 (n=149) and 2011 (n=133) and found an overall exposure rate to PRV of 24.1% (n=68); PRV was detected at two of three study sites. Two wild pigs had detectable antibody to B. abortus , and one had detectable antibody to PRRS. On average, 27% of wild pigs within a sounder were positive for PRV antibody, with 44% of the sounders (16/36) having at least one positive individual. These data highlight that wild pigs could carry pathogens that affect domestic livestock. Because the US is free of these pathogens in commercial livestock operations, continued surveillance and vaccination of domestic livestock are needed. Commercial livestock producers at the wildlife-livestock interface may benefit from spatial prioritization of risk zones to facilitate strategic control efforts.

  3. Prion-specific Hsp40 function: The role of the auxilin homolog Swa2

    PubMed Central

    Oliver, Emily E.; Troisi, Elizabeth M.; Hines, Justin K.

    2017-01-01

    ABSTRACT Yeast prions are protein-based genetic elements that propagate through cell populations via cytosolic transfer from mother to daughter cell. Molecular chaperone proteins including Hsp70, the Hsp40/J-protein Sis1, and Hsp104 are required for continued prion propagation, however the specific requirements of chaperone proteins differ for various prions. We recently reported that Swa2, the yeast homolog of the mammalian protein auxilin, is specifically required for the propagation of the prion [URE3].1 [URE3] propagation requires both a functional J-domain and the tetratricopeptide repeat (TPR) domain of Swa2, but does not require Swa2 clathrin binding. We concluded that the TPR domain determines the specificity of the genetic interaction between Swa2 and [URE3], and that this domain likely interacts with one or more proteins with a C-terminal EEVD motif. Here we extend that analysis to incorporate additional data that supports this hypothesis. We also present new data eliminating Hsp104 as the relevant Swa2 binding partner and discuss our findings in the context of other recent work involving Hsp90. Based on these findings, we propose a new model for Swa2's involvement in [URE3] propagation in which Swa2 and Hsp90 mediate the formation of a multi-protein complex that increases the number of sites available for Hsp104 disaggregation. PMID:28574745

  4. Seroepidemiology of Brucella abortus infection in bovids in Sri Lanka.

    PubMed

    Silva, I; Dangolla, A; Kulachelvy, K

    2000-07-03

    From 1992 to 1995, 0.15% (n=3916) of the bovids (cattle and buffalo) in Sri Lanka were sampled, using a multi-stage sampling procedure. Serum antibodies for Brucella abortus were detected using the indirect enzyme-linked immunosorbent assay (ELISA). The age, the agroecological zone and the management system practiced in the farms of the sampled bovids were studied as risk factors for seropositivity. The overall seroprevalence of brucellosis in cattle was 4.7% (n=3076) and 4.2% in buffaloes (n=840). Bovids that were over 3 years of age, from the dry zone (annual rainfall 20-35 in.), and reared under an extensive management system had higher odds of being seropositive. Bovids from the dry zone were at approximately six times higher odds of being seropositive even after controlling for the possible effects of age and management system. Approximately 75% of the seropositive males were from the dry zone. Most bovids (84%) from the dry zone in this study were reared under an extensive management system (free grazing) which allows unrestricted contact between animals. These factors may have contributed to the spread of brucellosis in bovids in the dry zone. This infection might be an important cause of abortion in bovids in Sri Lanka. However, there is also evidence of other causes for abortion, repeat breeding and retained placenta.

  5. Subcomponent vaccine based on CTA1-DD adjuvant with incorporated UreB class II peptides stimulates protective Helicobacter pylori immunity.

    PubMed

    Nedrud, John G; Bagheri, Nayer; Schön, Karin; Xin, Wei; Bergroth, Hilda; Eliasson, Dubravka Grdic; Lycke, Nils Y

    2013-01-01

    A mucosal vaccine against Helicobacter pylori infection could help prevent gastric cancers and peptic ulcers. While previous attempts to develop such a vaccine have largely failed because of the requirement for safe and effective adjuvants or large amounts of well defined antigens, we have taken a unique approach to combining our strong mucosal CTA1-DD adjuvant with selected peptides from urease B (UreB). The protective efficacy of the selected peptides together with cholera toxin (CT) was first confirmed. However, CT is a strong adjuvant that unfortunately is precluded from clinical use because of its toxicity. To circumvent this problem we have developed a derivative of CT, the CTA1-DD adjuvant, that has been found safe in non-human primates and equally effective compared to CT when used intranasally. We genetically fused the selected peptides into the CTA1-DD plasmid and found after intranasal immunizations of Balb/c mice using purified CTA1-DD with 3 copies of an H. pylori urease T cell epitope (CTA1-UreB3T-DD) that significant protection was stimulated against a live challenge infection. Protection was, however, weaker than with the gold standard, bacterial lysate+CT, but considering that we only used a single epitope in nanomolar amounts the results convey optimism. Protection was associated with enhanced Th1 and Th17 immunity, but immunizations in IL-17A-deficient mice revealed that IL-17 may not be essential for protection. Taken together, we have provided evidence for the rational design of an effective mucosal subcomponent vaccine against H. pylori infection based on well selected protective epitopes from relevant antigens incorporated into the CTA1-DD adjuvant platform.

  6. Generation of Cre-transgenic mice using Dlx1/Dlx2 enhancers and their characterization in GABAergic interneurons

    PubMed Central

    Potter, Gregory B.; Petryniak, Magdalena A.; Shevchenko, Eugenia; McKinsey, Gabriel L.; Ekker, Marc; Rubenstein, John L.R.

    2009-01-01

    DLX1 and DLX2 transcription factors are necessary for forebrain GABAergic neuron differentiation, migration, and survival. We generated transgenic mice that express Cre-recombinase under the control of two ultra-conserved DNA elements near the Dlx1&2 locus termed I12b and URE2. We show that Cre-recombinase is active in a “Dlx-pattern” in the embryonic forebrain of transgenic mice. I12b-Cre is more active than URE2-Cre in the medial ganglionic eminences and its derivatives. Fate-mapping of EGFP+ cells in adult Cre;Z/EG animals demonstrated that GABAergic neurons, but not glia, are labeled. Most NPY+, nNOS+, parvalbumin+, and somatostatin+ cells are marked by I12b-Cre in the cortex and hippocampus, while 25-40% of these interneuron subtypes are labeled by URE2-Cre. Labeling of neurons generated between E12.5 to E15.5 indicated differences in birth-dates of EGFP+ cells that populate the olfactory bulb, hippocampus, and cortex. Finally, we provide the first in vivo evidence that both I12b and URE2 are direct targets of DLX2 and require Dlx1 and Dlx2 expression for proper activity. PMID:19026749

  7. Crystallographic and kinetic study of riboflavin synthase from Brucella abortus, a chemotherapeutic target with an enhanced intrinsic flexibility

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Serer, María I.; Bonomi, Hernán R.; Guimarães, Beatriz G.

    This work reports crystal structures of trimeric riboflavin synthase from the pathogen B. abortus both as the apo protein and in complex with several ligands of interest. It is shown that ligand binding drives the assembly of the unique active site of the trimer, and these findings are complemented by a detailed kinetic study on this enzyme, in which marked inhibition by substrate and product was observed. Riboflavin synthase (RS) catalyzes the last step of riboflavin biosynthesis in microorganisms and plants, which corresponds to the dismutation of two molecules of 6,7-dimethyl-8-ribityllumazine to yield one molecule of riboflavin and one moleculemore » of 5-amino-6-ribitylamino-2,4(1H,3H)-pyrimidinedione. Owing to the absence of this enzyme in animals and the fact that most pathogenic bacteria show a strict dependence on riboflavin biosynthesis, RS has been proposed as a potential target for antimicrobial drug development. Eubacterial, fungal and plant RSs assemble as homotrimers lacking C{sub 3} symmetry. Each monomer can bind two substrate molecules, yet there is only one active site for the whole enzyme, which is located at the interface between two neighbouring chains. This work reports the crystallographic structure of RS from the pathogenic bacterium Brucella abortus (the aetiological agent of the disease brucellosis) in its apo form, in complex with riboflavin and in complex with two different product analogues, being the first time that the structure of an intact RS trimer with bound ligands has been solved. These crystal models support the hypothesis of enhanced flexibility in the particle and also highlight the role of the ligands in assembling the unique active site. Kinetic and binding studies were also performed to complement these findings. The structural and biochemical information generated may be useful for the rational design of novel RS inhibitors with antimicrobial activity.« less

  8. Evaluation of Brucella abortus Phosphoglucomutase (pgm) Mutant as a New Live Rough-Phenotype Vaccine

    PubMed Central

    Ugalde, Juan Esteban; Comerci, Diego José; Leguizamón, M. Susana; Ugalde, Rodolfo Augusto

    2003-01-01

    Brucella abortus S19 is the vaccine most frequently used against bovine brucellosis. Although it induces good protection levels, it cannot be administered to pregnant cattle, revaccination is not advised due to interference in the discrimination between infected and vaccinated animals during immune-screening procedures, and the vaccine is virulent for humans. Due to these reasons, there is a continuous search for new bovine vaccine candidates that may confer protection levels comparable to those conferred by S19 but without its disadvantages. A previous study characterized the phenotype associated with the phosphoglucomutase (pgm) gene disruption in Brucella abortus S2308, as well as the possible role for the smooth lipopolysaccharide (LPS) in virulence and intracellular multiplication in HeLa cells (J. E. Ugalde, C. Czibener, M. F. Feldman, and R. A. Ugalde, Infect. Immun. 68:5716-5723, 2000). In this report, we analyze the protection, proliferative response, and cytokine production induced in BALB/c mice by a Δpgm deletion strain. We show that this strain synthesizes O antigen with a size of approximately 45 kDa but is rough. This is due to the fact that the Δpgm strain is unable to assemble the O side chain in the complete LPS. Vaccination with the Δpgm strain induced protection levels comparable to those induced by S19 and generated a proliferative splenocyte response and a cytokine profile typical of a Th1 response. On the other hand, we were unable to detect a specific anti-O-antigen antibody response by using the fluorescence polarization assay. In view of these results, the possibility that the Δpgm mutant could be used as a vaccination strain is discussed. PMID:14573645

  9. Transcriptome Analysis of the Brucella abortus BvrR/BvrS Two-Component Regulatory System

    PubMed Central

    Viadas, Cristina; Rodríguez, María C.; Sangari, Felix J.; Gorvel, Jean-Pierre; García-Lobo, Juan M.; López-Goñi, Ignacio

    2010-01-01

    Background The two-component BvrR/BvrS system is essential for Brucella abortus virulence. It was shown previously that its dysfunction alters the expression of some major outer membrane proteins and the pattern of lipid A acylation. To determine the genes regulated by BvrR/BvrS, we performed a whole-genome microarray analysis using B. abortus RNA obtained from wild type and bvrR mutant cells grown in the same conditions. Methodology/Principal Findings A total of 127 differentially expressed genes were found: 83 were over expressed and 44 were less expressed in the bvrR mutant. Two operons, the phosphotransferase system and the maltose transport system, were down-regulated. Several genes involved in cell envelope or outer membrane biogenesis were differentially expressed: genes for outer membrane proteins (omp25a, omp25d), lipoproteins, LPS and fatty acid biosynthesis, stress response proteins, chaperones, flagellar genes, and twelve genes encoding ABC transport systems. Ten genes related with carbon metabolism (pckA and fumB among others) were up-regulated in the bvrR mutant, and denitrification genes (nirK, norC and nosZ) were also regulated. Notably, seven transcriptional regulators were affected, including VjbR, ExoR and OmpR that were less expressed in the bvrR mutant. Finally, the expression of eleven genes which have been previously related with Brucella virulence was also altered. Conclusions/Significance All these data corroborate the impact of BvrR/BvrS on cell envelope modulation, confirm that this system controls the carbon and nitrogen metabolism, and suggest a cross-talk among some regulators to adjust the Brucella physiology to the shift expected to occur during the transit from the extracellular to the intracellular niche. PMID:20422049

  10. Generation and characterization of β1,2-gluco-oligosaccharide probes from Brucella abortus cyclic β-glucan and their recognition by C-type lectins of the immune system

    PubMed Central

    Zhang, Hongtao; Palma, Angelina S; Zhang, Yibing; Childs, Robert A; Liu, Yan; Mitchell, Daniel A; Guidolin, Leticia S; Weigel, Wilfried; Mulloy, Barbara; Ciocchini, Andrés E; Feizi, Ten; Chai, Wengang

    2016-01-01

    The β1,2-glucans produced by bacteria are important in invasion, survival and immunomodulation in infected hosts be they mammals or plants. However, there has been a lack of information on proteins which recognize these molecules. This is partly due to the extremely limited availability of the sequence-defined oligosaccharides and derived probes for use in the study of their interactions. Here we have used the cyclic β1,2-glucan (CβG) of the bacterial pathogen Brucella abortus, after removal of succinyl side chains, to prepare linearized oligosaccharides which were used to generate microarrays. We describe optimized conditions for partial depolymerization of the cyclic glucan by acid hydrolysis and conversion of the β1,2-gluco-oligosaccharides, with degrees of polymerization 2–13, to neoglycolipids for the purpose of generating microarrays. By microarray analyses, we show that the C-type lectin receptor DC-SIGNR, like the closely related DC-SIGN we investigated earlier, binds to the β1,2-gluco-oligosaccharides, as does the soluble immune effector serum mannose-binding protein. Exploratory studies with DC-SIGN are suggestive of the recognition also of the intact CβG by this receptor. These findings open the way to unravelling mechanisms of immunomodulation mediated by β1,2-glucans in mammalian systems. PMID:27053576

  11. Immune responses and protection against experimental challenge after vaccination of bison with Brucella abortus strains RB51 or RB51 overexpressing superoxide dismutase and Glycosyltransferase genes

    USDA-ARS?s Scientific Manuscript database

    Vaccination is a tool that could be beneficial in managing the high prevalence of brucellosis in free-ranging bison in Yellowstone National Park. In this study, we characterized immunologic responses and protection against experimental challenge after vaccination of bison with Brucella abortus stra...

  12. Functional conservation of a forebrain enhancer from the elephant shark (Callorhinchus milii ) in zebrafish and mice.

    PubMed

    MacDonald, Ryan B; Debiais-Thibaud, Mélanie; Martin, Kyle; Poitras, Luc; Tay, Boon-Hui; Venkatesh, Byrappa; Ekker, Marc

    2010-05-26

    The phylogenetic position of the elephant shark (Callorhinchus milii ) is particularly relevant to study the evolution of genes and gene regulation in vertebrates. Here we examine the evolution of Dlx homeobox gene regulation during vertebrate embryonic development with a particular focus on the forebrain. We first identified the elephant shark sequence orthologous to the URE2 cis -regulatory element of the mouse Dlx1/Dlx2 locus (herein named CmURE2). We then conducted a comparative study of the sequence and enhancer activity of CmURE2 with that of orthologous regulatory sequences from zebrafish and mouse. The CmURE2 sequence shows a high percentage of identity with its mouse and zebrafish counterparts but is overall more similar to mouse URE2 (MmURE2) than to zebrafish URE2 (DrURE2). In transgenic zebrafish and mouse embryos, CmURE2 displayed enhancer activity in the forebrain that overlapped with that of DrURE2 and MmURE2. However, we detected notable differences in the activity of the three sequences in the diencephalon. Outside of the forebrain, CmURE2 shows enhancer activity in areas such as the pharyngeal arches and dorsal root ganglia where its' counterparts are also active. Our transgenic assays show that part of the URE2 enhancer activity is conserved throughout jawed vertebrates but also that new characteristics have evolved in the different groups. Our study demonstrates that the elephant shark is a useful outgroup to study the evolution of regulatory mechanisms in vertebrates and to address how changes in the sequence of cis -regulatory elements translate into changes in their regulatory activity.

  13. Subcomponent Vaccine Based on CTA1-DD Adjuvant with Incorporated UreB Class II Peptides Stimulates Protective Helicobacter pylori Immunity

    PubMed Central

    Nedrud, John G.; Bagheri, Nayer; Schön, Karin; Xin, Wei; Bergroth, Hilda; Eliasson, Dubravka Grdic; Lycke, Nils Y.

    2013-01-01

    A mucosal vaccine against Helicobacter pylori infection could help prevent gastric cancers and peptic ulcers. While previous attempts to develop such a vaccine have largely failed because of the requirement for safe and effective adjuvants or large amounts of well defined antigens, we have taken a unique approach to combining our strong mucosal CTA1-DD adjuvant with selected peptides from urease B (UreB). The protective efficacy of the selected peptides together with cholera toxin (CT) was first confirmed. However, CT is a strong adjuvant that unfortunately is precluded from clinical use because of its toxicity. To circumvent this problem we have developed a derivative of CT, the CTA1-DD adjuvant, that has been found safe in non-human primates and equally effective compared to CT when used intranasally. We genetically fused the selected peptides into the CTA1-DD plasmid and found after intranasal immunizations of Balb/c mice using purified CTA1-DD with 3 copies of an H. pylori urease T cell epitope (CTA1-UreB3T-DD) that significant protection was stimulated against a live challenge infection. Protection was, however, weaker than with the gold standard, bacterial lysate+CT, but considering that we only used a single epitope in nanomolar amounts the results convey optimism. Protection was associated with enhanced Th1 and Th17 immunity, but immunizations in IL-17A-deficient mice revealed that IL-17 may not be essential for protection. Taken together, we have provided evidence for the rational design of an effective mucosal subcomponent vaccine against H. pylori infection based on well selected protective epitopes from relevant antigens incorporated into the CTA1-DD adjuvant platform. PMID:24391754

  14. Brucellosis seroprevalence in Bali cattle with reproductive failure in South Sulawesi and Brucella abortus biovar 1 genotypes in the Eastern Indonesian archipelago.

    PubMed

    Muflihanah, Hanah; Hatta, Mochammad; Rood, Ente; Scheelbeek, Pauline; Abdoel, Theresia H; Smits, Henk L

    2013-11-26

    Brucellosis is a major cause of infertility and reproductive failure in livestock. While cattle in the Eastern Indonesian archipelago suffers from reproductive problems information on bovine brucellosis in the region is fragmentary. The control of brucellosis requires a major and prolonged effort and confirmation of the infection by isolation with detailed knowledge of the spread of the infection is essential when planning a control program. Serological investigation of Brucella infection in beef cattle tended under extensive farming conditions revealed a high seroprevalence (19.3%; 95% CI, 17-22) in the compliment fixation tests. The results of a rapid and simple field test correlated well with the Rose Bengal test (kappa, 0.917) and indicated an acceptable sensitivity (87.5%) and specificity (98.1%) compared with the complement fixation test. Reproductive failure was reported for 39.0% of the cows with a loss of calves due to abortion or early death amounting to 19.3%. Past reproductive failure did not, however, correlate with seropositivity in the complement fixation test (RP = 1.21; P = 0.847). B. abortus biovar 1 was freshly isolated from the hygromas of two cows and together with thirty banked isolates collected since 1990 from different parts of Sulawesi and Timor eight related genotypes could be distinguished with one genotype being identical to that of an isolate (BfR91) from Switzerland. The Indonesian genotypes formed together with BfR91 and one African and one North American isolate a distinct branch on the B. abortus biovar 1 dendogram. Bovine brucellosis appears to be widespread in the Eastern Indonesian archipelago and calls for urgent intervention. The fresh isolation of the pathogen together with the observed high seroprevalence demonstrates the presence and frequent exposure of cattle in the area to the pathogen. The application of a rapid and simple field test for brucellosis could be very useful for the quick screening of cattle at the pen side.

  15. Structural, functional and immunogenic insights on Cu,Zn superoxide dismutase pathogenic virulence factors from Neisseria meningitidis and Brucella abortus

    DOE PAGES

    Pratt, Ashley J.; DiDonato, Michael; Shin, David S.; ...

    2015-10-12

    Bacterial pathogens Neisseria meningitidis and Brucella abortus pose threats to human and animal health worldwide, causing meningococcal disease and brucellosis, respectively. Mortality from acute N. meningitidis infections remains high despite antibiotics, and brucellosis presents alimentary and health consequences. Superoxide dismutases are master regulators of reactive oxygen, general pathogenicity factors and therefore therapeutic targets. Cu,Zn superoxide dismutases (SODs) localized to the periplasm promote survival by detoxifying superoxide radicals generated by major host antimicrobial immune responses. We discovered that passive immunization with an antibody directed at N. meningitidis SOD (NmSOD) was protective in a mouse infection model. To define the relevant atomicmore » details and solution assembly states of this important virulence factor, we report high-resolution and X-ray scattering analyses of NmSOD and SOD from B. abortus (BaSOD). The NmSOD structures revealed an auxiliary tetrahedral Cu-binding site bridging the dimer interface; mutational analyses suggested that this metal site contributes to protein stability, with implications for bacterial defense mechanisms. Biochemical and structural analyses informed us about electrostatic substrate guidance, dimer assembly and an exposed C-terminal epitope in the NmSOD dimer. In contrast, the monomeric BaSOD structure provided insights for extending immunogenic peptide epitopes derived from the protein. These collective results reveal unique contributions of SOD to pathogenic virulence, refine predictive motifs for distinguishing SOD classes and suggest general targets for anti-bacterial immune responses. The identified functional contributions, motifs, and targets distinguishing bacterial and eukaryotic SOD assemblies presented here provide a foundation for efforts to develop SOD-specific inhibitors or vaccines against these harmful pathogens. IMPORTANCE By protecting microbes against reactive oxygen

  16. Structural, Functional, and Immunogenic Insights on Cu,Zn Superoxide Dismutase Pathogenic Virulence Factors from Neisseria meningitidis and Brucella abortus

    PubMed Central

    Pratt, Ashley J.; DiDonato, Michael; Shin, David S.; Cabelli, Diane E.; Bruns, Cami K.; Belzer, Carol A.; Gorringe, Andrew R.; Langford, Paul R.; Tabatabai, Louisa B.; Kroll, J. Simon; Tainer, John A.

    2015-01-01

    ABSTRACT Bacterial pathogens Neisseria meningitidis and Brucella abortus pose threats to human and animal health worldwide, causing meningococcal disease and brucellosis, respectively. Mortality from acute N. meningitidis infections remains high despite antibiotics, and brucellosis presents alimentary and health consequences. Superoxide dismutases are master regulators of reactive oxygen and general pathogenicity factors and are therefore therapeutic targets. Cu,Zn superoxide dismutases (SODs) localized to the periplasm promote survival by detoxifying superoxide radicals generated by major host antimicrobial immune responses. We discovered that passive immunization with an antibody directed at N. meningitidis SOD (NmSOD) was protective in a mouse infection model. To define the relevant atomic details and solution assembly states of this important virulence factor, we report high-resolution and X-ray scattering analyses of NmSOD and of SOD from B. abortus (BaSOD). The NmSOD structures revealed an auxiliary tetrahedral Cu-binding site bridging the dimer interface; mutational analyses suggested that this metal site contributes to protein stability, with implications for bacterial defense mechanisms. Biochemical and structural analyses informed us about electrostatic substrate guidance, dimer assembly, and an exposed C-terminal epitope in the NmSOD dimer. In contrast, the monomeric BaSOD structure provided insights for extending immunogenic peptide epitopes derived from the protein. These collective results reveal unique contributions of SOD to pathogenic virulence, refine predictive motifs for distinguishing SOD classes, and suggest general targets for antibacterial immune responses. The identified functional contributions, motifs, and targets distinguishing bacterial and eukaryotic SOD assemblies presented here provide a foundation for efforts to develop SOD-specific inhibitors of or vaccines against these harmful pathogens. IMPORTANCE By protecting microbes against

  17. Structural, Functional, and Immunogenic Insights on Cu,Zn Superoxide Dismutase Pathogenic Virulence Factors from Neisseria meningitidis and Brucella abortus.

    PubMed

    Pratt, Ashley J; DiDonato, Michael; Shin, David S; Cabelli, Diane E; Bruns, Cami K; Belzer, Carol A; Gorringe, Andrew R; Langford, Paul R; Tabatabai, Louisa B; Kroll, J Simon; Tainer, John A; Getzoff, Elizabeth D

    2015-12-01

    Bacterial pathogens Neisseria meningitidis and Brucella abortus pose threats to human and animal health worldwide, causing meningococcal disease and brucellosis, respectively. Mortality from acute N. meningitidis infections remains high despite antibiotics, and brucellosis presents alimentary and health consequences. Superoxide dismutases are master regulators of reactive oxygen and general pathogenicity factors and are therefore therapeutic targets. Cu,Zn superoxide dismutases (SODs) localized to the periplasm promote survival by detoxifying superoxide radicals generated by major host antimicrobial immune responses. We discovered that passive immunization with an antibody directed at N. meningitidis SOD (NmSOD) was protective in a mouse infection model. To define the relevant atomic details and solution assembly states of this important virulence factor, we report high-resolution and X-ray scattering analyses of NmSOD and of SOD from B. abortus (BaSOD). The NmSOD structures revealed an auxiliary tetrahedral Cu-binding site bridging the dimer interface; mutational analyses suggested that this metal site contributes to protein stability, with implications for bacterial defense mechanisms. Biochemical and structural analyses informed us about electrostatic substrate guidance, dimer assembly, and an exposed C-terminal epitope in the NmSOD dimer. In contrast, the monomeric BaSOD structure provided insights for extending immunogenic peptide epitopes derived from the protein. These collective results reveal unique contributions of SOD to pathogenic virulence, refine predictive motifs for distinguishing SOD classes, and suggest general targets for antibacterial immune responses. The identified functional contributions, motifs, and targets distinguishing bacterial and eukaryotic SOD assemblies presented here provide a foundation for efforts to develop SOD-specific inhibitors of or vaccines against these harmful pathogens. By protecting microbes against reactive oxygen

  18. Structural, functional and immunogenic insights on Cu,Zn superoxide dismutase pathogenic virulence factors from Neisseria meningitidis and Brucella abortus

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Pratt, Ashley J.; DiDonato, Michael; Shin, David S.

    Bacterial pathogens Neisseria meningitidis and Brucella abortus pose threats to human and animal health worldwide, causing meningococcal disease and brucellosis, respectively. Mortality from acute N. meningitidis infections remains high despite antibiotics, and brucellosis presents alimentary and health consequences. Superoxide dismutases are master regulators of reactive oxygen, general pathogenicity factors and therefore therapeutic targets. Cu,Zn superoxide dismutases (SODs) localized to the periplasm promote survival by detoxifying superoxide radicals generated by major host antimicrobial immune responses. We discovered that passive immunization with an antibody directed at N. meningitidis SOD (NmSOD) was protective in a mouse infection model. To define the relevant atomicmore » details and solution assembly states of this important virulence factor, we report high-resolution and X-ray scattering analyses of NmSOD and SOD from B. abortus (BaSOD). The NmSOD structures revealed an auxiliary tetrahedral Cu-binding site bridging the dimer interface; mutational analyses suggested that this metal site contributes to protein stability, with implications for bacterial defense mechanisms. Biochemical and structural analyses informed us about electrostatic substrate guidance, dimer assembly and an exposed C-terminal epitope in the NmSOD dimer. In contrast, the monomeric BaSOD structure provided insights for extending immunogenic peptide epitopes derived from the protein. These collective results reveal unique contributions of SOD to pathogenic virulence, refine predictive motifs for distinguishing SOD classes and suggest general targets for anti-bacterial immune responses. The identified functional contributions, motifs, and targets distinguishing bacterial and eukaryotic SOD assemblies presented here provide a foundation for efforts to develop SOD-specific inhibitors or vaccines against these harmful pathogens. IMPORTANCE By protecting microbes against reactive oxygen

  19. First Report of Orchitis in Man Caused by Brucella abortus Biovar 1 in Ecuador

    PubMed Central

    Ron-Román, Jorge; Saegerman, Claude; Minda-Aluisa, Elizabeth; Benítez-Ortíz, Washington; Brandt, Jef; Douce, Richard

    2012-01-01

    We present a 44-year-old man from a rural community in northern Ecuador who worked on a cattle farm where he was involved with primary veterinary care, including assistance during births (or calving) and placenta retention and artificial insemination, with minimal precautions. In September of 2009, quite abruptly, he developed asthenia and hypersomnia without any apparent cause or symptoms like fever, chills, or night sweats. On November 14, 2009, he suffered from pain and edema in the right testicle that coincided with pain in the abdomen. Clinical, serological, and bacteriological investigations confirmed the first case of unilateral orchitis in man in Ecuador caused by Brucella abortus biovar 1. Because brucellosis is a neglected disease, special attention should be given to it in the training of medical and veterinary students. PMID:22826490

  20. Evaluation of Brucella abortus strain RB51 and strain 19 in pronghorn antelope

    USGS Publications Warehouse

    Elzer, P.H.; Smith, J.; Roffe, T.; Kreeger, T.; Edwards, J.; Davis, D.

    2002-01-01

    Free-roaming elk and bison in the Greater Yellowstone Area remain the only wildlife reservoirs for Brucella abortus in the United States, and the large number of animals and a lack of holding facilities make it unreasonable to individually vaccinate each animal. Therefore, oral delivery is being proposed as a possible option to vaccinate these wild ungulates. One of the main problems associated with oral vaccination is the potential exposure of nontarget species to the vaccines. The purpose of this study was to determine the effects of two Brucella vaccines, strain 19 (S19) and the rough strain RB51 (SRB51), in pregnant pronghorn antelope. We conclude that S19 and SRB51 rarely colonize maternal and fetal tissues of pregnant pronghorn and were not associated with fetal death. Oral delivery of either vaccine at this dose appears to be nonhazardous to pregnant pronghorn.

  1. Brucella abortus Strain 2308 Wisconsin Genome: Importance of the Definition of Reference Strains

    PubMed Central

    Suárez-Esquivel, Marcela; Ruiz-Villalobos, Nazareth; Castillo-Zeledón, Amanda; Jiménez-Rojas, César; Roop II, R. Martin; Comerci, Diego J.; Barquero-Calvo, Elías; Chacón-Díaz, Carlos; Caswell, Clayton C.; Baker, Kate S.; Chaves-Olarte, Esteban; Thomson, Nicholas R.; Moreno, Edgardo; Letesson, Jean J.; De Bolle, Xavier; Guzmán-Verri, Caterina

    2016-01-01

    Brucellosis is a bacterial infectious disease affecting a wide range of mammals and a neglected zoonosis caused by species of the genetically homogenous genus Brucella. As in most studies on bacterial diseases, research in brucellosis is carried out by using reference strains as canonical models to understand the mechanisms underlying host pathogen interactions. We performed whole genome sequencing analysis of the reference strain B. abortus 2308 routinely used in our laboratory, including manual curated annotation accessible as an editable version through a link at https://en.wikipedia.org/wiki/Brucella#Genomics. Comparison of this genome with two publically available 2308 genomes showed significant differences, particularly indels related to insertional elements, suggesting variability related to the transposition of these elements within the same strain. Considering the outcome of high resolution genomic techniques in the bacteriology field, the conventional concept of strain definition needs to be revised. PMID:27746773

  2. Immune responses of bison and efficacy after booster vaccination with Brucella abortus strain RB51.

    PubMed

    Olsen, S C; McGill, J L; Sacco, R E; Hennager, S G

    2015-04-01

    Thirty-one bison heifers were randomly assigned to receive saline or a single vaccination with 10(10) CFU of Brucella abortus strain RB51. Some vaccinated bison were randomly selected for booster vaccination with RB51 at 11 months after the initial vaccination. Mean antibody responses to RB51 were greater (P < 0.05) in vaccinated bison after initial and booster vaccination than in nonvaccinated bison. The proliferative responses by peripheral blood mononuclear cells (PBMC) from the vaccinated bison were greater (P < 0.05) than those in the nonvaccinated bison at 16 and 24 weeks after the initial vaccination but not after the booster vaccination. The relative gene expression of gamma interferon (IFN-γ) was increased (P < 0.05) in the RB51-vaccinated bison at 8, 16, and 24 weeks after the initial vaccination and at 8 weeks after the booster vaccination. The vaccinated bison had greater (P < 0.05) in vitro production of IFN-γ at all sampling times, greater interleukin-1β (IL-1β) production in various samplings after the initial and booster vaccinations, and greater IL-6 production at one sampling time after the booster vaccination. Between 170 and 180 days of gestation, the bison were intraconjunctivally challenged with approximately 1 × 10(7) CFU of B. abortus strain 2308. The incidences of abortion and infection were greater (P < 0.05) in the nonvaccinated bison after experimental challenge than in the bison receiving either vaccination treatment. Booster-vaccinated, but not single-vaccinated bison, had a reduced (P < 0.05) incidence of infection in fetal tissues and maternal tissues compared to that in the controls. Compared to the nonvaccinated bison, both vaccination treatments lowered the colonization (measured as the CFU/g of tissue) of Brucella organisms in all tissues, except in retropharyngeal and supramammary lymph nodes. Our study suggests that RB51 booster vaccination is an effective vaccination strategy for enhancing herd immunity against

  3. Immune Responses of Bison and Efficacy after Booster Vaccination with Brucella abortus Strain RB51

    PubMed Central

    McGill, J. L.; Sacco, R. E.; Hennager, S. G.

    2015-01-01

    Thirty-one bison heifers were randomly assigned to receive saline or a single vaccination with 1010 CFU of Brucella abortus strain RB51. Some vaccinated bison were randomly selected for booster vaccination with RB51 at 11 months after the initial vaccination. Mean antibody responses to RB51 were greater (P < 0.05) in vaccinated bison after initial and booster vaccination than in nonvaccinated bison. The proliferative responses by peripheral blood mononuclear cells (PBMC) from the vaccinated bison were greater (P < 0.05) than those in the nonvaccinated bison at 16 and 24 weeks after the initial vaccination but not after the booster vaccination. The relative gene expression of gamma interferon (IFN-γ) was increased (P < 0.05) in the RB51-vaccinated bison at 8, 16, and 24 weeks after the initial vaccination and at 8 weeks after the booster vaccination. The vaccinated bison had greater (P < 0.05) in vitro production of IFN-γ at all sampling times, greater interleukin-1β (IL-1β) production in various samplings after the initial and booster vaccinations, and greater IL-6 production at one sampling time after the booster vaccination. Between 170 and 180 days of gestation, the bison were intraconjunctivally challenged with approximately 1 × 107 CFU of B. abortus strain 2308. The incidences of abortion and infection were greater (P < 0.05) in the nonvaccinated bison after experimental challenge than in the bison receiving either vaccination treatment. Booster-vaccinated, but not single-vaccinated bison, had a reduced (P < 0.05) incidence of infection in fetal tissues and maternal tissues compared to that in the controls. Compared to the nonvaccinated bison, both vaccination treatments lowered the colonization (measured as the CFU/g of tissue) of Brucella organisms in all tissues, except in retropharyngeal and supramammary lymph nodes. Our study suggests that RB51 booster vaccination is an effective vaccination strategy for enhancing herd immunity against brucellosis in

  4. Factors Required for Activation of Urease as a Virulence Determinant in Cryptococcus neoformans

    PubMed Central

    Singh, Arpita; Panting, Robert J.; Varma, Ashok; Saijo, Tomomi; Waldron, Kevin J.; Jong, Ambrose; Ngamskulrungroj, Popchai; Chang, Yun C.; Rutherford, Julian C.; Kwon-Chung, Kyung J.

    2013-01-01

    ABSTRACT Urease in Cryptococcus neoformans plays an important role in fungal dissemination to the brain and causing meningoencephalitis. Although urea is not required for synthesis of apourease encoded by URE1, the available nitrogen source affected the expression of URE1 as well as the level of the enzyme activity. Activation of the apoenzyme requires three accessory proteins, Ure4, Ure6, and Ure7, which are homologs of the bacterial urease accessory proteins UreD, UreF, and UreG, respectively. A yeast two-hybrid assay showed positive interaction of Ure1 with the three accessory proteins encoded by URE4, URE6, and URE7. Metalloproteomic analysis of cryptococcal lysates using inductively coupled plasma mass spectrometry (ICP-MS) and a biochemical assay of urease activity showed that, as in many other organisms, urease is a metallocentric enzyme that requires nickel transported by Nic1 for its catalytic activity. The Ure7 accessory protein (bacterial UreG homolog) binds nickel likely via its conserved histidine-rich domain and appears to be responsible for the incorporation of Ni2+ into the apourease. Although the cryptococcal genome lacks the bacterial UreE homolog, Ure7 appears to combine the functions of bacterial UreE and UreG, thus making this pathogen more similar to that seen with the plant system. Brain invasion by the ure1, ure7, and nic1 mutant strains that lack urease activity was significantly less effective in a mouse model. This indicated that an activated urease and not the Ure1 protein was responsible for enhancement of brain invasion and that the factors required for urease activation in C. neoformans resemble those of plants more than those of bacteria. PMID:23653445

  5. Mass vaccination as a complementary tool in the control of a severe outbreak of bovine brucellosis due to Brucella abortus in Extremadura, Spain.

    PubMed

    Sanz, Cristina; Sáez, José Luis; Alvarez, Julio; Cortés, María; Pereira, Gema; Reyes, Aurelia; Rubio, Félix; Martín, Javier; García, Nerea; Domínguez, Lucas; Hermoso-de-Mendoza, María; Hermoso-de-Mendoza, Javier

    2010-11-01

    We report the evolution of an outbreak of bovine brucellosis (Brucella abortus) in the region of Extremadura (Spain) involving more than 1000 herds and nearly 40,000 animals. S19 vaccination of young cattle combined with a test and slaughter strategy did not result in a rapid decrease in herd prevalence and animal incidence; these parameters showed a constant decreasing trend only when a combination of restriction of cattle movements, increased test frequency, S19 vaccination and mass RB51 vaccination (with yearly revaccinations) were applied to all susceptible populations. These measures were applied for 5 years; abortions following RB51 vaccination of pregnant cows were limited to the first inoculation and the involvement of the vaccine strain could only be demonstrated in 78 out of 897 abortions. Our results demonstrate the usefulness - and lack of significant side effects - of RB51 mass vaccination as a complementary tool to control bovine brucellosis outbreaks in areas where the disease cannot be contained using more conservative approaches. Copyright © 2010 Elsevier B.V. All rights reserved.

  6. An oral vaccine based on U-Omp19 induces protection against B. abortus mucosal challenge by inducing an adaptive IL-17 immune response in mice.

    PubMed

    Pasquevich, Karina A; Ibañez, Andrés E; Coria, Lorena M; García Samartino, Clara; Estein, Silvia M; Zwerdling, Astrid; Barrionuevo, Paula; Oliveira, Fernanda S; Seither, Christine; Warzecha, Heribert; Oliveira, Sergio C; Giambartolomei, Guillermo H; Cassataro, Juliana

    2011-01-14

    As Brucella infections occur mainly through mucosal surfaces, the development of mucosal administered vaccines could be radical for the control of brucellosis. In this work we evaluated the potential of Brucella abortus 19 kDa outer membrane protein (U-Omp19) as an edible subunit vaccine against brucellosis. We investigated the protective immune response elicited against oral B. abortus infection after vaccination of mice with leaves from transgenic plants expressing U-Omp19; or with plant-made or E. coli-made purified U-Omp19. All tested U-Omp19 formulations induced protection against Brucella when orally administered without the need of adjuvants. U-Omp19 also induced protection against a systemic challenge when parenterally administered. This built-in adjuvant ability of U-Omp19 was independent of TLR4 and could be explained at least in part by its capability to activate dendritic cells in vivo. While unadjuvanted U-Omp19 intraperitoneally administered induced a specific Th1 response, following U-Omp19 oral delivery a mixed specific Th1-Th17 response was induced. Depletion of CD4(+) T cells in mice orally vaccinated with U-Omp19 resulted in a loss of the elicited protection, indicating that this cell type mediates immune protection. The role of IL-17 against Brucella infection has never been explored. In this study, we determined that if IL-17A was neutralized in vivo during the challenge period, the mucosal U-Omp19 vaccine did not confer mucosal protection. On the contrary, IL-17A neutralization during the infection did not influence at all the subsistence and growth of this bacterium in PBS-immunized mice. All together, our results indicate that an oral unadjuvanted vaccine based on U-Omp19 induces protection against a mucosal challenge with Brucella abortus by inducing an adaptive IL-17 immune response. They also indicate different and important new aspects i) IL-17 does not contribute to reduce the bacterial burden in non vaccinated mice and ii) IL-17 plays

  7. Brucella abortus Choloylglycine Hydrolase Affects Cell Envelope Composition and Host Cell Internalization

    PubMed Central

    Marchesini, María Inés; Connolly, Joseph; Delpino, María Victoria; Baldi, Pablo C.; Mujer, Cesar V.; DelVecchio, Vito G.; Comerci, Diego J.

    2011-01-01

    Choloylglycine hydrolase (CGH, E.C. 3.5.1.24) is a conjugated bile salt hydrolase that catalyses the hydrolysis of the amide bond in conjugated bile acids. Bile salt hydrolases are expressed by gastrointestinal bacteria, and they presumably decrease the toxicity of host's conjugated bile salts. Brucella species are the causative agents of brucellosis, a disease affecting livestock and humans. CGH confers Brucella the ability to deconjugate and resist the antimicrobial action of bile salts, contributing to the establishment of a successful infection through the oral route in mice. Additionally, cgh-deletion mutant was also attenuated in intraperitoneally inoculated mice, which suggests that CGH may play a role during systemic infection other than hydrolyzing conjugated bile acids. To understand the role CGH plays in B. abortus virulence, we infected phagocytic and epithelial cells with a cgh-deletion mutant (Δcgh) and found that it is defective in the internalization process. This defect along with the increased resistance of Δcgh to the antimicrobial action of polymyxin B, prompted an analysis of the cell envelope of this mutant. Two-dimensional electrophoretic profiles of Δcgh cell envelope-associated proteins showed an altered expression of Omp2b and different members of the Omp25/31 family. These results were confirmed by Western blot analysis with monoclonal antibodies. Altogether, the results indicate that Brucella CGH not only participates in deconjugation of bile salts but also affects overall membrane composition and host cell internalization. PMID:22174816

  8. Brucella abortus choloylglycine hydrolase affects cell envelope composition and host cell internalization.

    PubMed

    Marchesini, María Inés; Connolly, Joseph; Delpino, María Victoria; Baldi, Pablo C; Mujer, Cesar V; DelVecchio, Vito G; Comerci, Diego J

    2011-01-01

    Choloylglycine hydrolase (CGH, E.C. 3.5.1.24) is a conjugated bile salt hydrolase that catalyses the hydrolysis of the amide bond in conjugated bile acids. Bile salt hydrolases are expressed by gastrointestinal bacteria, and they presumably decrease the toxicity of host's conjugated bile salts. Brucella species are the causative agents of brucellosis, a disease affecting livestock and humans. CGH confers Brucella the ability to deconjugate and resist the antimicrobial action of bile salts, contributing to the establishment of a successful infection through the oral route in mice. Additionally, cgh-deletion mutant was also attenuated in intraperitoneally inoculated mice, which suggests that CGH may play a role during systemic infection other than hydrolyzing conjugated bile acids. To understand the role CGH plays in B. abortus virulence, we infected phagocytic and epithelial cells with a cgh-deletion mutant (Δcgh) and found that it is defective in the internalization process. This defect along with the increased resistance of Δcgh to the antimicrobial action of polymyxin B, prompted an analysis of the cell envelope of this mutant. Two-dimensional electrophoretic profiles of Δcgh cell envelope-associated proteins showed an altered expression of Omp2b and different members of the Omp25/31 family. These results were confirmed by Western blot analysis with monoclonal antibodies. Altogether, the results indicate that Brucella CGH not only participates in deconjugation of bile salts but also affects overall membrane composition and host cell internalization.

  9. Methods for Real-Time PCR-Based Diagnosis of Chlamydia pneumoniae, Chlamydia psittaci, and Chlamydia abortus Infections in an Opened Molecular Diagnostic Platform.

    PubMed

    Opota, Onya; Brouillet, René; Greub, Gilbert; Jaton, Katia

    2017-01-01

    The advances in molecular biology of the last decades have dramatically improved the field of diagnostic bacteriology. In particular, PCR-based technologies have impacted the diagnosis of infections caused by obligate intracellular bacteria such as pathogens from the Chlamydiacae family. Here, we describe a real-time PCR-based method using the Taqman technology for the diagnosis of Chlamydia pneumoniae, Chlamydia psittaci, and Chlamydia abortus infection. The method presented here can be applied to various clinical samples and can be adapted on opened molecular diagnostic platforms.

  10. MyD88 and STING Signaling Pathways Are Required for IRF3-Mediated IFN-β Induction in Response to Brucella abortus Infection

    PubMed Central

    de Almeida, Leonardo A.; Carvalho, Natalia B.; Oliveira, Fernanda S.; Lacerda, Thais L. S.; Vasconcelos, Anilton C.; Nogueira, Lucas; Bafica, Andre; Silva, Aristóbolo M.; Oliveira, Sergio C.

    2011-01-01

    Type I interferons (IFNs) are cytokines that orchestrate diverse immune responses to viral and bacterial infections. Although typically considered to be most important molecules in response to viruses, type I IFNs are also induced by most, if not all, bacterial pathogens. In this study, we addressed the role of type I IFN signaling during Brucella abortus infection, a facultative intracellular bacterial pathogen that causes abortion in domestic animals and undulant fever in humans. Herein, we have shown that B. abortus induced IFN-β in macrophages and splenocytes. Further, IFN-β induction by Brucella was mediated by IRF3 signaling pathway and activates IFN-stimulated genes via STAT1 phosphorylation. In addition, IFN-β expression induced by Brucella is independent of TLRs and TRIF signaling but MyD88-dependent, a pathway not yet described for Gram-negative bacteria. Furthermore, we have identified Brucella DNA as the major bacterial component to induce IFN-β and our study revealed that this molecule operates through a mechanism dependent on RNA polymerase III to be sensed probably by an unknown receptor via the adaptor molecule STING. Finally, we have demonstrated that IFN-αβR KO mice are more resistant to infection suggesting that type I IFN signaling is detrimental to host control of Brucella. This resistance phenotype is accompanied by increased IFN-γ and NO production by IFN-αβR KO spleen cells and reduced apoptosis. PMID:21829705

  11. MLVA and MLST typing of Brucella from Qinghai, China.

    PubMed

    Ma, Jun-Ying; Wang, Hu; Zhang, Xue-Fei; Xu, Li-Qing; Hu, Gui-Ying; Jiang, Hai; Zhao, Fang; Zhao, Hong-Yan; Piao, Dong-Ri; Qin, Yu-Min; Cui, Bu-Yun; Lin, Gong-Hua

    2016-04-13

    The Qinghai-Tibet Plateau (QTP) of China is an extensive pastoral and semi-pastoral area, and because of poverty and bad hygiene conditions, Brucella is highly prevalent in this region. In order to adequately prevent this disease in the QTP region it is important to determine the identity of Brucella species that caused the infection. A total of 65 Brucella isolates were obtained from human, livestock and wild animals in Qinghai, a Chinese province in east of the QTP. Two molecular typing methods, MLVA (multi-locus variable-number tandem-repeat analysis) and MLST (multi locus sequence typing) were used to identify the species and genotypes of these isolates. Both MLVA and MLST typing methods classified the 65 isolates into three species, B. melitensis, B. abortus and B. suis, which included 60, 4 and 1 isolates respectively. The MLVA method uniquely detected 34 (Bm01 ~ Bm34), 3 (Ba01 ~ Ba03), and 1 (Bs01) MLVA-16 genotypes for B. melitensis, B. abortus and B. suis, respectively. However, none of these genotypes exactly matched any of the genotypes in the Brucella2012 MLVA database. The MLST method identified five known ST types: ST7 and ST8 (B. melitensis), ST2 and ST5 (B. abortus), and ST14 (B. suis). We also detected a strain with a mutant type (3-2-3-2-?-5-3-8-2) of ST8 (3-2-3-2-1-5-3-8-2). Extensive genotype-sharing events could be observed among isolates from different host species. There were at least three Brucella (B. melitensis, B. abortus and B. suis) species in Qinghai, of which B. melitensis was the predominant species in the area examined. The Brucella population in Qinghai was very different from other regions of the world, possibly owing to the unique geographical characteristics such as extremely high altitude in QTP. There were extensive genotype-sharing events between isolates obtained from humans and other animals. Yaks, sheep and blue sheep were important zoonotic reservoirs of brucellosis causing species found in humans.

  12. The Dutch Brucella abortus monitoring programme for cattle: the impact of false-positive serological reactions and comparison of serological tests.

    PubMed

    Emmerzaal, A; de Wit, J J; Dijkstra, Th; Bakker, D; van Zijderveld, F G

    2002-02-01

    The Dutch national Brucella abortus eradication programme for cattle started in 1959. Sporadic cases occurred yearly until 1995; the last infected herd was culled in 1996. In August 1999 the Netherlands was declared officially free of bovine brucellosis by the European Union. Before 1999, the programme to monitor the official Brucella-free status of bovine herds was primarily based on periodical testing of dairy herds with the milk ring test (MRT) and serological testing of all animals older than 1 year of age from non-dairy herds, using the micro-agglutination test (MAT) as screening test. In addition, serum samples of cattle that aborted were tested with the MAT. The high number of false positive reactions in both tests and the serum agglutination test (SAT) and complement fixation test (CFT) used for confirmation seemed to result in unnecessary blockade of herds, subsequent testing and slaughter of animals. For this reason, a validation study was performed in which three indirect enzyme-linked immunosorbent assays (ELISAs), the CFT and the SAT were compared using a panel of sera from brucellosis-free cattle, sera from experimentally infected cattle, and sera from cattle experimentally infected with bacteria which are known to induce cross-reactive antibodies (Pasteurella, Salmonella, Yersinia, and Escherichia). Moreover, four ELISAs and the MRT were compared using a panel of 1000 bulk milk samples from Brucella-free herds and 12 milk samples from Brucella abortus- infected cattle. It is concluded that the ELISA obtained from ID-Lelystad is the most suitable test to monitor the brucelosis free status of herds because it gives rise to fewer false-positive reactions than the SAT.

  13. New Features in the Lipid A Structure of Brucella suis and Brucella abortus Lipopolysaccharide

    NASA Astrophysics Data System (ADS)

    Casabuono, Adriana C.; Czibener, Cecilia; Del Giudice, Mariela G.; Valguarnera, Ezequiel; Ugalde, Juan E.; Couto, Alicia S.

    2017-12-01

    Brucellaceae are Gram-negative bacteria that cause brucellosis, one of the most distributed worldwide zoonosis, transmitted to humans by contact with either infected animals or their products. The lipopolysaccharide exposed on the cell surface has been intensively studied and is considered a major virulence factor of Brucella. In the last years, structural studies allowed the determination of new structures in the core oligosaccharide and the O-antigen of this lipopolysaccharide. In this work, we have reinvestigated the lipid A structure isolated from B. suis and B. abortus lipopolysaccharides. A detailed study by MALDI-TOF mass spectrometry in the positive and negative ion modes of the lipid A moieties purified from both species was performed. Interestingly, a new feature was detected: the presence of a pyrophosphorylethanolamine residue substituting the backbone. LID-MS/MS analysis of some of the detected ions allowed assurance that the Lipid A structure composed by the diGlcN3N disaccharide, mainly hexa-acylated and penta-acylated, bearing one phosphate and one pyrophosphorylethanolamine residue. [Figure not available: see fulltext.

  14. New Features in the Lipid A Structure of Brucella suis and Brucella abortus Lipopolysaccharide.

    PubMed

    Casabuono, Adriana C; Czibener, Cecilia; Del Giudice, Mariela G; Valguarnera, Ezequiel; Ugalde, Juan E; Couto, Alicia S

    2017-12-01

    Brucellaceae are Gram-negative bacteria that cause brucellosis, one of the most distributed worldwide zoonosis, transmitted to humans by contact with either infected animals or their products. The lipopolysaccharide exposed on the cell surface has been intensively studied and is considered a major virulence factor of Brucella. In the last years, structural studies allowed the determination of new structures in the core oligosaccharide and the O-antigen of this lipopolysaccharide. In this work, we have reinvestigated the lipid A structure isolated from B. suis and B. abortus lipopolysaccharides. A detailed study by MALDI-TOF mass spectrometry in the positive and negative ion modes of the lipid A moieties purified from both species was performed. Interestingly, a new feature was detected: the presence of a pyrophosphorylethanolamine residue substituting the backbone. LID-MS/MS analysis of some of the detected ions allowed assurance that the Lipid A structure composed by the diGlcN3N disaccharide, mainly hexa-acylated and penta-acylated, bearing one phosphate and one pyrophosphorylethanolamine residue. Graphical abstract ᅟ.

  15. Express immunochromatographic detection of antibodies against Brucella abortus in cattle sera based on quantitative photometric registration and modulated cut-off level.

    PubMed

    Sotnikov, Dmitriy V; Byzova, Nadezhda A; Zherdev, Anatoly V; Eskendirova, Saule Z; Baltin, Kairat K; Mukanov, Kasim K; Ramankulov, Erlan M; Sadykhov, Elchin G; Dzantiev, Boris B

    2015-01-01

    An immunochromatographic test system was developed for rapid detection of the levels of specific IgG antibodies to Brucella abortus lipopolysaccharide, as a tool for diagnosis of brucellosis in cattle. The pilot test strips were examined using blood sera from sick (78 samples) and healthy (35 samples) cows. The results obtained by immunochromatographic assay, using a portable optical densitometer for digital video detection, correlate well with the results obtained by immunoenzyme assay and are in agreement with the results of the disease diagnosis. The new test system allows detection of antibodies within 10 min and can be proposed as an alternative to the methods available for serodiagnosis of brucellosis.

  16. Gln3p and Nil1p regulation of invertase activity and SUC2 expression in Saccharomyces cerevisiae.

    PubMed

    Oliveira, Edna Maria Morais; Mansure, José João; Bon, Elba Pinto da Silva

    2005-04-01

    In Saccharomyces cerevisiae, sensing and signalling pathways regulate gene expression in response to quality of carbon and nitrogen sources. One such system, the target of rapamycin (Tor) proteins, senses nutrients and uses the GATA activators Gln3p and Nil1p to regulate translation in response to low-quality carbon and nitrogen. The signal transduction, triggered in response to nitrogen nutrition that is sensed by the Tor proteins, operates via a regulatory pathway involving the cytoplasmic factor Ure2p. When carbon and nitrogen are abundant, the phosphorylated Ure2p anchors the also phosphorylated Gln3p and Nil1p in the cytoplasm. Upon a shift from high- to low-quality nitrogen or treatment with rapamycin all three proteins are dephosphorylated, causing Gln3p and Nil1p to enter the nucleus and promote transcription. The genes that code for yeast periplasmic enzymes with nutritional roles would be obvious targets for regulation by the sensing and signalling pathways that respond to quality of carbon and nitrogen sources. Indeed, previous results from our laboratory had shown that the GATA factors Gln3p, Nil1p, Dal80p, Nil2p and also the protein Ure2 regulate the expression of asparaginase II, coded by ASP3. We also had observed that the activity levels of the also periplasmic invertase, coded by SUC2, were 6-fold lower in ure2 mutant cells in comparison to wild-type cells collected at stationary phase. These results suggested similarities between the signalling pathways regulating the expression of ASP3 and SUC2. In the present work we showed that invertase levels displayed by the single nil1 and gln3 and by the double gln3nil1 mutant cells, cultivated in a sucrose-ammonium medium and collected at the exponential phase, were 6-, 10- and 60-fold higher, respectively, in comparison to their wild-type counterparts. RT-PCR data of SUC2 expression in the double-mutant cells indicated a 10-fold increase in the mRNA(SUC2) levels.

  17. Outer membrane proteins from rough strains of four Brucella species.

    PubMed Central

    Santos, J M; Verstreate, D R; Perera, V Y; Winter, A J

    1984-01-01

    Outer membrane proteins from 15 rough strains of Brucella abortus, B. ovis, B. canis, and B. melitensis were extracted with a dipolar detergent, and outer membrane proteins from selected strains were purified by anion exchange chromatography and gel filtration (Verstreate et al., Infect. Immun. 35:979-989, 1982). Outer membrane proteins produced two types of profiles on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. One type, demonstrated by B. abortus, B. ovis, and B. canis strains, contained the three predominant protein groups present in smooth B. abortus strains (Verstreate et al., Infect. Immun. 35:979-989, 1982): groups 1, 2 (porin [Douglas et al., Infect. Immun. 44:16-21]), and 3. B. melitensis strains demonstrated the second profile type, in which there was an additional band between groups 1 and 2. The relative proportion of porin was considerably lower in B. ovis, B. canis, and B. melitensis than in B. abortus. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis profiles could be used to distinguish B. abortus and B. melitensis from each other and from B. canis and B. ovis. The amino acid compositions of groups 2 and 3 from rough strains of B. abortus, B. canis, and B. melitensis were similar to those of corresponding proteins from smooth B. abortus strains. Zwittergent-soluble fractions from most rough strains contained antigen [b], which cross-reacted with group 2 from smooth B. abortus strains, and antigens [c] and [d], which cross-reacted with group 3 from smooth B. abortus strains. Antigen [a], shared by groups 2 and 3 (D. R. Verstreate and A. J. Winter, Infect. Immun. 46:182-187, 1984), was detected in most rough strains. None of these antigens were related to either rough or smooth lipopolysaccharide. Images PMID:6480106

  18. Cryptococcus gattii urease as a virulence factor and the relevance of enzymatic activity in cryptococcosis pathogenesis.

    PubMed

    Feder, Vanessa; Kmetzsch, Lívia; Staats, Charley Christian; Vidal-Figueiredo, Natalia; Ligabue-Braun, Rodrigo; Carlini, Célia Regina; Vainstein, Marilene Henning

    2015-04-01

    Ureases (EC 3.5.1.5) are Ni(2+) -dependent metalloenzymes produced by plants, fungi and bacteria that hydrolyze urea to produce ammonia and CO2 . The insertion of nickel atoms into the apo-urease is better characterized in bacteria, and requires at least three accessory proteins: UreD, UreF, and UreG. Our group has demonstrated that ureases possess ureolytic activity-independent biological properties that could contribute to the pathogenicity of urease-producing microorganisms. The presence of urease in pathogenic bacteria strongly correlates with pathogenesis in some human diseases. Some medically important fungi also produce urease, including Cryptococcus neoformans and Cryptococcus gattii. C. gattii is an etiological agent of cryptococcosis, most often affecting immunocompetent individuals. The cryptococcal urease might play an important role in pathogenesis. It has been proposed that ammonia produced via urease action might damage the host endothelium, which would enable yeast transmigration towards the central nervous system. To analyze the role of urease as a virulence factor in C. gattii, we constructed knockout mutants for the structural urease-coding gene URE1 and for genes that code the accessory proteins Ure4 and Ure6. All knockout mutants showed reduced multiplication within macrophages. In intranasally infected mice, the ure1Δ (lacking urease protein) and ure4Δ (enzymatically inactive apo-urease) mutants caused reduced blood burdens and a delayed time of death, whereas the ure6Δ (enzymatically inactive apo-urease) mutant showed time and dose dependency with regard to fungal burden. Our results suggest that C. gattii urease plays an important role in virulence, in part possibly through enzyme activity-independent mechanism(s). © 2015 FEBS.

  19. Prepatellar bursitis due to Brucella abortus: case report and analysis of the local immune response.

    PubMed

    Wallach, Jorge C; Delpino, M Victoria; Scian, Romina; Deodato, Bettina; Fossati, Carlos A; Baldi, Pablo C

    2010-12-01

    A case of prepatellar bursitis in a man with chronic brucellosis is presented. Brucella abortus biotype 1 was isolated from the abundant yellowish fluid obtained from the bursa. Clinical and epidemiological data did not suggest a direct inoculation of the agent in the bursa. However, the patient mentioned occasional local trauma due to recreational sports, which may have constituted a predisposing factor. As determined by ELISA, there were higher levels of IgG against Brucella LPS and cytosolic proteins detected in the patient's bursal synovial fluid when compared with serum. Levels of proinflammatory cytokines (tumour necrosis factor alpha, interleukin 1 beta, gamma interferon, interleukin 8 and MCP-1) were higher than in synovial fluids obtained from patients with rheumatoid arthritis and a patient with septic arthritis, and a zymographic analysis revealed a gelatinase of about 92 kDa. These findings indicate that it may be possible to diagnose brucellar bursitis by measuring specific antibodies in the bursal synovial fluid. In addition, our findings suggest a role of increased local levels of proinflammatory cytokines and gelatinases in the inflammatory manifestations of brucellar bursitis.

  20. Epidemiology of bovine brucellosis in Costa Rica: Lessons learned from failures in the control of the disease.

    PubMed

    Hernández-Mora, Gabriela; Ruiz-Villalobos, Nazareth; Bonilla-Montoya, Roberto; Romero-Zúniga, Juan-José; Jiménez-Arias, Julio; González-Barrientos, Rocío; Barquero-Calvo, Elías; Chacón-Díaz, Carlos; Rojas, Norman; Chaves-Olarte, Esteban; Guzmán-Verri, Caterina; Moreno, Edgardo

    2017-01-01

    Brucellosis, caused by Brucella abortus is a major disease of cattle and a zoonosis. In order to estimate the bovine brucellosis prevalence in Costa Rica (CR), a total 765 herds (13078 bovines) from six regions of CR were randomly sampled during 2012-2013. A non-random sample of 7907 herds (532199 bovines) of the six regions, arriving for diagnoses during 2014-2016 to the Costa Rican Animal Health Service was also studied. The prevalence estimated by Rose Bengal test (RBT) ranged from 10.5%-11.4%; alternatively, the prevalence estimated by testing the RBT positives in iELISA, ranged from 4.1%-6.0%, respectively. However, cattle in CR are not vaccinated with B. abortus S19 but with RB51 (vaccination coverage close to 11%), and under these conditions the RBT displays 99% specificity and 99% sensitivity. Therefore, the RBT herd depicted in the random analysis stands as a feasible assessment and then, the recommended value in case of planning an eradication program in CR. Studies of three decades reveled that bovine brucellosis prevalence has increased in CR. B. abortus was identified by biochemical and molecular studies as the etiological agent of bovine brucellosis. Multiple locus variable-number tandem repeat analysis-16 revealed four B. abortus clusters. Cluster one and three are intertwined with isolates from other countries, while clusters two and four have only representatives from CR. Cluster one is widely distributed in all regions of the country and may be the primary B. abortus source. The other clusters seem to be restricted to specific areas in CR. The implications of our findings, in relation to the control of the disease in CR, are critically discussed.

  1. Utility of p57 immunohistochemistry in differentiating between complete mole, partial mole & non-molar or hydropic abortus.

    PubMed

    Samadder, Abhimanyu; Kar, Rakhee

    2017-01-01

    There is considerable inter-observer variability in the diagnosis of molar pregnancies by histomorphological examination of products of conception (POC). The p57KIP2 gene is paternally imprinted and expressed from the maternal allele. On immunohistochemistry (IHC) with p57, complete mole (CM) shows absent staining whereas hydropic abortus (HA) and partial mole (PM) show positive staining. This study was undertaken to evaluate the role of p57 IHC along with histomorphology in differentiating between CM, PM and non-molar or HA. This was a cross-sectional study over a period of three and a half years on archival material. Detailed histomorphological review along with p57 IHC was carried out in 28 diagnosed cases (23 CM, 4 PM and 1 molar pregnancy not categorized) and 25 controls of four normal placentas and 21 POC (8 non-hydropic and 13 HA). In 14.8 per cent (4/27) cases, there was discordance in accurate subtyping of molar pregnancy. One case of CM showed inconsistent IHC pattern. In 15.4 per cent (2/13) HA, molar pregnancy was final diagnosis. After final review, there were 25 CM, five PM, 22 non-molar controls including 10 HA and one not assigned (PM/HA). IHC with p57 was negative in 96 per cent CM and positive in 100 and 95 per cent PM and non-molar controls, respectively. This study showed that negative p57KIP2 immunostaining reliably identified CM and could be used in association with the histological findings to distinguish CM from its mimics.

  2. Apoprotein isolation and activation, and vibrational structure of the Helicobacter mustelae iron urease

    PubMed Central

    Carter, Eric L.; Proshlyakov, Denis A.; Hausinger, Robert P.

    2011-01-01

    The micro-aerophilic pathogen Helicobacter mustelae synthesizes an oxygen-labile, iron-containing urease (UreA2B2) in addition to its standard nickel-containing enzyme (UreAB). An apoprotein form of the iron urease was prepared from ureA2B2-expressing recombinant Escherichia coli cells that were grown in minimal medium. Temperature-dependent circular dichroism measurements of holoprotein and apoprotein demonstrate an enhancement of thermal stability associated with the UreA2B2 metallocenter. In parallel to the situation reported for nickel activation of the standard urease apoprotein, incubation of UreA2B2 apoprotein with ferrous ions and bicarbonate generated urease activity in a portion of the nascent active sites. In addition, ferrous ions were shown to be capable of reductively activating the oxidized metallocenter. Resonance Raman spectra of the inactive, aerobically-purified UreA2B2 holoprotein exhibit vibrations at 495 cm−1 and 784 cm−1, consistent with νs and νas modes of an Fe(III)-O-Fe(III) center; these modes undergo downshifts upon binding of urea and were unaffected by changes in pH. The low-frequency mode also exhibits an isotopic shift from 497 to 476 cm−1 upon 16O/18O bulk water isotope substitution. Expression of subunits of the conventional nickel-containing Klebsiella aerogenes urease in cells grown in rich medium without nickel resulted in iron incorporation into a portion of the protein. The inactive iron-loaded species exhibited a UV-visible spectrum similar to oxidized UreA2B2 and was capable of being reductively activated under anoxic conditions. Results from these studies more clearly define the formation and unique properties of the iron urease metallocenter. PMID:22196017

  3. Biosafety of parenteral Brucella abortus RB51 vaccine in bison calves

    USGS Publications Warehouse

    Roffe, T.J.; Olsen, S.C.; Gidlewski, T.; Jensen, A.E.; Palmer, M.V.; Huber, R.

    1999-01-01

    Vaccination is considered among the primary management tools for reducing brucellosis prevalence in Greater Yellowstone Area (GYA) ungulates. Before their use, however, vaccine safety and efficacy must be demonstrated. Twenty-seven female bison (Bison bison) calves (approx 5 months old) were vaccinated with Brucella abortus Strain RB51 (1.5 x 1010 colony forming units [CFU], subcutaneously) as part of routine management. We assessed the persistence, pathology, shedding, and transmission associated with RB51 by serial necropsy, bacteriology, histopathology, and serology of 20 of these 27 vaccinated calves, and RB51 serology of 10 nonvaccinated, commingling adult females. With the exception of 1 calf, RB51 dot-blot titers at necropsy were <1:80. Strain RB51 was cultured from lymph nodes in 4 of 4 calves at 14 weeks postvaccination (PV), 4 of 4 calves at 18 weeks PV, 1 of 4 calves at 22 weeks PV, 3 of 4 at 26 weeks PV, and 0 of 4 calves at 30 weeks PV. No gross lesions were observed. Mild histologic changes occurred only in a few draining lymph nodes early in sampling. Adverse clinical effects were not observed in vaccinates. Swabs from nasopharynx, conjunctiva, rectum, and vagina were uniformly culture negative for RB51. Strain RB51 dot-blot assays of bison cows were negative at a 1:20 dilution at 26 weeks PV. Our results suggest that RB51 persists longer in bison calves than in domestic cattle and is systemically distributed within lymphatic tissues. However, bison apparently clear the RB51 vaccine strain without shedding, transmission, or significant adverse reactions.

  4. Epidemiology of bovine brucellosis in Costa Rica: Lessons learned from failures in the control of the disease

    PubMed Central

    Hernández-Mora, Gabriela; Ruiz-Villalobos, Nazareth; Bonilla-Montoya, Roberto; Romero-Zúniga, Juan-José; Jiménez-Arias, Julio; González-Barrientos, Rocío; Barquero-Calvo, Elías; Chacón-Díaz, Carlos; Rojas, Norman; Chaves-Olarte, Esteban; Guzmán-Verri, Caterina

    2017-01-01

    Brucellosis, caused by Brucella abortus is a major disease of cattle and a zoonosis. In order to estimate the bovine brucellosis prevalence in Costa Rica (CR), a total 765 herds (13078 bovines) from six regions of CR were randomly sampled during 2012–2013. A non-random sample of 7907 herds (532199 bovines) of the six regions, arriving for diagnoses during 2014–2016 to the Costa Rican Animal Health Service was also studied. The prevalence estimated by Rose Bengal test (RBT) ranged from 10.5%-11.4%; alternatively, the prevalence estimated by testing the RBT positives in iELISA, ranged from 4.1%-6.0%, respectively. However, cattle in CR are not vaccinated with B. abortus S19 but with RB51 (vaccination coverage close to 11%), and under these conditions the RBT displays 99% specificity and 99% sensitivity. Therefore, the RBT herd depicted in the random analysis stands as a feasible assessment and then, the recommended value in case of planning an eradication program in CR. Studies of three decades reveled that bovine brucellosis prevalence has increased in CR. B. abortus was identified by biochemical and molecular studies as the etiological agent of bovine brucellosis. Multiple locus variable-number tandem repeat analysis-16 revealed four B. abortus clusters. Cluster one and three are intertwined with isolates from other countries, while clusters two and four have only representatives from CR. Cluster one is widely distributed in all regions of the country and may be the primary B. abortus source. The other clusters seem to be restricted to specific areas in CR. The implications of our findings, in relation to the control of the disease in CR, are critically discussed. PMID:28797045

  5. Metabolic Control of Virulence Genes in Brucella abortus: HutC Coordinates virB Expression and the Histidine Utilization Pathway by Direct Binding to Both Promoters ▿ †

    PubMed Central

    Sieira, Rodrigo; Arocena, Gastón M.; Bukata, Lucas; Comerci, Diego J.; Ugalde, Rodolfo A.

    2010-01-01

    Type IV secretion systems (T4SS) are multicomponent machineries involved in the translocation of effector molecules across the bacterial cell envelope. The virB operon of Brucella abortus codes for a T4SS that is essential for virulence and intracellular multiplication of the bacterium in the host. Previous studies showed that the virB operon of B. abortus is tightly regulated within the host cells. In order to identify factors implicated in the control of virB expression, we searched for proteins of Brucella that directly bind to the virB promoter (PvirB). Using different procedures, we isolated a 27-kDa protein that binds specifically to PvirB. This protein was identified as HutC, the transcriptional repressor of the histidine utilization (hut) genes. Analyses of virB and hut promoter activity revealed that HutC exerts two different roles: it acts as a coactivator of transcription of the virB operon, whereas it represses the hut genes. Such activities were observed both intracellularly and in bacteria incubated under conditions that resemble the intracellular environment. Electrophoresis mobility shift assays (EMSA) and DNase I footprinting experiments revealed the structure, affinity, and localization of the HutC-binding sites and supported the regulatory role of HutC in both hut and virB promoters. Taken together, these results indicate that Brucella coopted the function of HutC to coordinate the Hut pathway with transcriptional regulation of the virB genes, probably as a way to sense its own metabolic state and develop adaptive responses to overcome intracellular host defenses. PMID:19854911

  6. Brucella abortus Depends on Pyruvate Phosphate Dikinase and Malic Enzyme but Not on Fbp and GlpX Fructose-1,6-Bisphosphatases for Full Virulence in Laboratory Models

    PubMed Central

    Zúñiga-Ripa, Amaia; Barbier, Thibault; Conde-Álvarez, Raquel; Martínez-Gómez, Estrella; Palacios-Chaves, Leyre; Gil-Ramírez, Yolanda; Grilló, María Jesús; Letesson, Jean-Jacques; Iriarte, Maite

    2014-01-01

    The brucellae are the etiological agents of brucellosis, a worldwide-distributed zoonosis. These bacteria are facultative intracellular parasites and thus are able to adjust their metabolism to the extra- and intracellular environments encountered during an infectious cycle. However, this aspect of Brucella biology is imperfectly understood, and the nutrients available in the intracellular niche are unknown. Here, we investigated the central pathways of C metabolism used by Brucella abortus by deleting the putative fructose-1,6-bisphosphatase (fbp and glpX), phosphoenolpyruvate carboxykinase (pckA), pyruvate phosphate dikinase (ppdK), and malic enzyme (mae) genes. In gluconeogenic but not in rich media, growth of ΔppdK and Δmae mutants was severely impaired and growth of the double Δfbp-ΔglpX mutant was reduced. In macrophages, only the ΔppdK and Δmae mutants showed reduced multiplication, and studies with the ΔppdK mutant confirmed that it reached the replicative niche. Similarly, only the ΔppdK and Δmae mutants were attenuated in mice, the former being cleared by week 10 and the latter persisting longer than 12 weeks. We also investigated the glyoxylate cycle. Although aceA (isocitrate lyase) promoter activity was enhanced in rich medium, aceA disruption had no effect in vitro or on multiplication in macrophages or mouse spleens. The results suggest that B. abortus grows intracellularly using a limited supply of 6-C (and 5-C) sugars that is compensated by glutamate and possibly other amino acids entering the Krebs cycle without a critical role of the glyoxylate shunt. PMID:24936050

  7. Magnitude and Determinants of the Ratio between Prevalence of Low Vision and Blindness in Rapid Assessment of Avoidable Blindness Surveys.

    PubMed

    Kaphle, Dinesh; Lewallen, Susan

    2017-10-01

    To determine the magnitude and determinants of the ratio between prevalence of low vision and prevalence of blindness in rapid assessment of avoidable blindness (RAAB) surveys globally. Standard RAAB reports were downloaded from the repository or requested from principal investigators. Potential predictor variables included prevalence of uncorrected refractive error (URE) as well as gross domestic product (GDP) per capita, health expenditure per capita of the country across World Bank regions. Univariate and multivariate linear regression were used to investigate the correlation between potential predictor variables and the ratio. The results of 94 surveys from 43 countries showed that the ratio ranged from 1.35 in Mozambique to 11.03 in India with a median value of 3.90 (Interquartile range 3.06;5.38). Univariate regression analysis showed that prevalence of URE (p = 0.04), logarithm of GDP per capita (p = 0.01) and logarithm of health expenditure per capita (p = 0.03) were significantly associated with the higher ratio. However, only prevalence of URE was found to be significant in multivariate regression analysis (p = 0.03). There is a wide variation in the ratio of the prevalence of low vision to the prevalence of blindness. Eye care service utilization indicators such as the prevalence of URE may explain some of the variation across the regions.

  8. Regional alternative transportation evaluation report - region 2

    DOT National Transportation Integrated Search

    2012-03-01

    The U.S. Fish and Wildlife Service (FWS) and the U.S. Department of Transportation (DOT) Volpe : Center (Volpe Center) conducted a regional alternative transportation evaluation (RATE) in Region 2, : which is comprised of Arizona, Oklahoma, New Mexic...

  9. Role of Helicobacter pylori methionine sulfoxide reductase in urease maturation

    PubMed Central

    Kuhns, Lisa G.; Mahawar, Manish; Sharp, Joshua S.; Benoit, Stéphane; Maier, Robert J.

    2014-01-01

    The persistence of the gastric pathogen Helicobacter pylori is due in part to urease and Msr (methionine sulfoxide reductase). Upon exposure to relatively mild (21% partial pressure of O2) oxidative stress, a Δmsr mutant showed both decreased urease specific activity in cell-free extracts and decreased nickel associated with the partially purified urease fraction as compared with the parent strain, yet urease apoprotein levels were the same for the Δmsr and wild-type extracts. Urease activity of the Δmsr mutant was not significantly different from the wild-type upon non-stress microaerobic incubation of strains. Urease maturation occurs through nickel mobilization via a suite of known accessory proteins, one being the GTPase UreG. Treatment of UreG with H2O2 resulted in oxidation of MS-identified methionine residues and loss of up to 70% of its GTPase activity. Incubation of pure H2O2-treated UreG with Msr led to reductive repair of nine methionine residues and recovery of up to full enzyme activity. Binding of Msr to both oxidized and non-oxidized UreG was observed by cross-linking. Therefore we conclude Msr aids the survival of H. pylori in part by ensuring continual UreG-mediated urease maturation under stress conditions. PMID:23181726

  10. Localization of antigen-specific lymphocytes following lymph node challenge.

    PubMed Central

    Liu, H; Splitter, G A

    1986-01-01

    The effect of subcutaneous injections of Brucella abortus strain 19 antigen on the specific localization of autologous lymphocytes in the regional nodes of calves was analysed by fluorescent labelling and flow cytometry. Both in vitro and in vivo FITC labelling of lymphocytes indicated the preferential migration of lymphocytes from a previously challenged lymph node to a recently challenged lymph node. However, lymphocytes from a lymph node challenged with B. abortus failed to localize preferentially in a lymph node challenged with a control antigen, Listeria monocytogenes. Lymph node cells, enriched for T lymphocytes and isolated from primary stimulated or secondary challenged B. abortus lymph nodes, could proliferate when cultured with autologous antigen-pulsed macrophages. The kinetics of [3H]thymidine incorporation in lymphocytes from secondarily challenged lymph nodes occurred earlier and to a greater extent when compared with lymphocytes from primary challenged lymph nodes. Our data show that the accumulation of B. abortus-specific lymphocytes in secondarily challenged lymph nodes is increased by the presence of the specific antigen. Images Figure 4 PMID:2426183

  11. Protein Tunnels: The Case of Urease Accessory Proteins.

    PubMed

    Musiani, Francesco; Gioia, Dario; Masetti, Matteo; Falchi, Federico; Cavalli, Andrea; Recanatini, Maurizio; Ciurli, Stefano

    2017-05-09

    Transition metals are both essential micronutrients and limited in environmental availability. The Ni(II)-dependent urease protein, the most efficient enzyme known to date, is a paradigm for studying the strategies that cells use to handle an essential, yet toxic, metal ion. Urease is a virulence factor of several human pathogens, in addition to decreasing the efficiency of soil organic nitrogen fertilization. Ni(II) insertion in the urease active site is performed through the action of three essential accessory proteins: UreD, UreF, and UreG. The crystal structure of the UreD-UreF-UreG complex from the human pathogen Helicobacter pylori (HpUreDFG) revealed the presence of tunnels that cross the entire length of both UreF and UreD, potentially able to deliver Ni(II) ions from UreG to apo-urease. Atomistic molecular dynamics simulations performed on the HpUreDFG complex in explicit solvent and at physiological ionic conditions demonstrate the stability of these protein tunnels in solution and provide insights on the trafficking of water molecules inside the tunnels. The presence of different alternative routes across the identified tunnels for Ni(II) ions, water molecules, and carbonate ions, all involved in urease activation, is highlighted here, and their potential role in the urease activation mechanism is discussed.

  12. Influence of uncorrected refractive error and unmet refractive error on visual impairment in a Brazilian population.

    PubMed

    Ferraz, Fabio H; Corrente, José E; Opromolla, Paula; Schellini, Silvana A

    2014-06-25

    The World Health Organization (WHO) definitions of blindness and visual impairment are widely based on best-corrected visual acuity excluding uncorrected refractive errors (URE) as a visual impairment cause. Recently, URE was included as a cause of visual impairment, thus emphasizing the burden of visual impairment due to refractive error (RE) worldwide is substantially higher. The purpose of the present study is to determine the reversal of visual impairment and blindness in the population correcting RE and possible associations between RE and individual characteristics. A cross-sectional study was conducted in nine counties of the western region of state of São Paulo, using systematic and random sampling of households between March 2004 and July 2005. Individuals aged more than 1 year old were included and were evaluated for demographic data, eye complaints, history, and eye exam, including no corrected visual acuity (NCVA), best corrected vision acuity (BCVA), automatic and manual refractive examination. The definition adopted for URE was applied to individuals with NCVA > 0.15 logMAR and BCVA ≤ 0.15 logMAR after refractive correction and unmet refractive error (UREN), individuals who had visual impairment or blindness (NCVA > 0.5 logMAR) and BCVA ≤ 0.5 logMAR after optical correction. A total of 70.2% of subjects had normal NCVA. URE was detected in 13.8%. Prevalence of 4.6% of optically reversible low vision and 1.8% of blindness reversible by optical correction were found. UREN was detected in 6.5% of individuals, more frequently observed in women over the age of 50 and in higher RE carriers. Visual impairment related to eye diseases is not reversible with spectacles. Using multivariate analysis, associations between URE and UREN with regard to sex, age and RE was observed. RE is an important cause of reversible blindness and low vision in the Brazilian population.

  13. [Mechanism of anti-Helicobacter pylori urease activity of patchouli alcohol].

    PubMed

    Lian, Da-Wei; Xu, Yi-Fei; Ren, Wen-Kang; Fu, Li-Jun; Fan, Ping-Long; Cao, Hong-Ying; Huang, Ping

    2017-02-01

    To investigate the effect of patchouli alcohol on inhibiting Helicobater pylori urease activity, and its effect on expression levels of related genes, and lay the foundation for further research on the effect of patchouli alcohol on H. pylori colonization and infection. H. pyloriwas cultured and identified by gram staining, rapid urease test (RUT) and PCR method. Then agar dilution method was used to detect the bacterial survival after 1 h intervention by different concentrations of patchouli alcoholin the acidic (pH 5.3) and neutral (pH 7.0) conditions; berthelot method was used to detect urease activity and RT-qPCR method was used to detect the expression changes of ureA, ureB, ureE, ureH, ureI, and nixA related urease genes. The results showed that the survival rate of H. pyloriwas not significantly changed but the urease activity was obviously decreased after intervention by different concentrations of patchouli alcohol; meanwhile, the expression levels of ureA, ureB, ureE, ureH, ureI, and nixA were decreased to different degrees. Therefore, patchouli alcohol could inhibit H. pylori urease activity in both acidic and neutral conditions, and the mechanism may be related to down-regulation of urease gene expression. Copyright© by the Chinese Pharmaceutical Association.

  14. Helicobacter pylori Infection in Rural and Urban Dyspeptic Patients from Venezuela

    PubMed Central

    Contreras, Monica; Fernández-Delgado, Milagro; Reyes, Nelson; García-Amado, María Alexandra; Rojas, Héctor; Michelangeli, Fabian

    2015-01-01

    The goal of this work was to assess the Helicobacter pylori prevalence in a rural mestizo population and compare it to an urban population from Venezuela. The study was performed in gastric juice samples of 71 dyspeptic patients from Caracas (urban) and 39 from Tucupita (rural), in the Orinoco Delta region. Helicobacter pylori was detected by amplification of 16S rRNA, glmM, and ureA genes in 55.0% patients from urban and 87.2% from rural populations. cagA was found positive in 51% and 62% urban and rural patients, respectively. Non-H. pylori Helicobacter species were not detected in the urban population, but was found in 7.7% of patients in the rural study site. Frequency values of the 16S rRNA, glmM, and ureA genes were higher in the rural population. The odds ratio for each gene was 15.18 for 16S rRNA, 2.34 for glmM, 2.89 for ureA, and 1.53 cagA, showing significant differences except for cagA when gene frequency was compared in both populations. These results demonstrate a higher frequency of H. pylori and gastric non-H. pylori Helicobacter infection in a rural mestizo population with low hygienic standards as compared with city dwellers, representing a potential risk for the development of gastroduodenal diseases. PMID:26195456

  15. WrpA Is an Atypical Flavodoxin Family Protein under Regulatory Control of the Brucella abortus General Stress Response System

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Herrou, Julien; Czyż, Daniel M.; Willett, Jonathan W.

    ABSTRACT The general stress response (GSR) system of the intracellular pathogenBrucella abortuscontrols the transcription of approximately 100 genes in response to a range of stress cues. The core genetic regulatory components of the GSR are required forB. abortussurvival under nonoptimal growth conditionsin vitroand for maintenance of chronic infection in anin vivomouse model. The functions of the majority of the genes in the GSR transcriptional regulon remain undefined.bab1_1070is among the most highly regulated genes in this regulon: its transcription is activated 20- to 30-fold by the GSR system under oxidative conditionsin vitro. We have solved crystal structures of Bab1_1070 and demonstratemore » that it forms a homotetrameric complex that resembles those of WrbA-type NADH:quinone oxidoreductases, which are members of the flavodoxin protein family. However,B. abortusWrbA-relatedprotein (WrpA) does not bind flavin cofactors with a high affinity and does not function as an NADH:quinone oxidoreductasein vitro. Soaking crystals with flavin mononucleotide (FMN) revealed a likely low-affinity binding site adjacent to the canonical WrbA flavin binding site. Deletion ofwrpA(ΔwrpA) does not compromise cell survival under acute oxidative stressin vitroor attenuate infection in cell-based or mouse models. However, a ΔwrpAstrain does elicit increased splenomegaly in a mouse model, suggesting that WrpA modulatesB. abortusinteraction with its mammalian host. Despite high structural homology with canonical WrbA proteins, we propose thatB. abortusWrpA represents a functionally distinct member of the diverse flavodoxin family. IMPORTANCEBrucella abortusis an etiological agent of brucellosis, which is among the most common zoonotic diseases worldwide. The general stress response (GSR) regulatory system ofB. abortuscontrols the transcription of approximately 100 genes and is required for maintenance of chronic infection in a murine model; the majority of GSR-regulated genes

  16. Functional expression of a heterologous nickel-dependent, ATP-independent urease in Saccharomyces cerevisiae.

    PubMed

    Milne, N; Luttik, M A H; Cueto Rojas, H F; Wahl, A; van Maris, A J A; Pronk, J T; Daran, J M

    2015-07-01

    In microbial processes for production of proteins, biomass and nitrogen-containing commodity chemicals, ATP requirements for nitrogen assimilation affect product yields on the energy producing substrate. In Saccharomyces cerevisiae, a current host for heterologous protein production and potential platform for production of nitrogen-containing chemicals, uptake and assimilation of ammonium requires 1 ATP per incorporated NH3. Urea assimilation by this yeast is more energy efficient but still requires 0.5 ATP per NH3 produced. To decrease ATP costs for nitrogen assimilation, the S. cerevisiae gene encoding ATP-dependent urease (DUR1,2) was replaced by a Schizosaccharomyces pombe gene encoding ATP-independent urease (ure2), along with its accessory genes ureD, ureF and ureG. Since S. pombe ure2 is a Ni(2+)-dependent enzyme and Saccharomyces cerevisiae does not express native Ni(2+)-dependent enzymes, the S. pombe high-affinity nickel-transporter gene (nic1) was also expressed. Expression of the S. pombe genes into dur1,2Δ S. cerevisiae yielded an in vitro ATP-independent urease activity of 0.44±0.01 µmol min(-1) mg protein(-1) and restored growth on urea as sole nitrogen source. Functional expression of the Nic1 transporter was essential for growth on urea at low Ni(2+) concentrations. The maximum specific growth rates of the engineered strain on urea and ammonium were lower than those of a DUR1,2 reference strain. In glucose-limited chemostat cultures with urea as nitrogen source, the engineered strain exhibited an increased release of ammonia and reduced nitrogen content of the biomass. Our results indicate a new strategy for improving yeast-based production of nitrogen-containing chemicals and demonstrate that Ni(2+)-dependent enzymes can be functionally expressed in S. cerevisiae. Copyright © 2015 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.

  17. Chlamydia abortus Pmp18.1 Induces IL-1β Secretion by TLR4 Activation through the MyD88, NF-κB, and Caspase-1 Signaling Pathways

    PubMed Central

    Pan, Qing; Zhang, Qiang; Chu, Jun; Pais, Roshan; Liu, Shanshan; He, Cheng; Eko, Francis O.

    2017-01-01

    The polymorphic membrane protein D (Pmp18D) is a 160-kDa outer membrane protein that is conserved and plays an important role in Chlamydia abortus pathogenesis. We have identified an N-terminal fragment of Pmp18D (designated Pmp18.1) as a possible subunit vaccine antigen. In this study, we evaluated the vaccine potential of Pmp18.1 by investigating its ability to induce innate immune responses in dendritic cells and the signaling pathway(s) involved in rPmp18.1-induced IL-1β secretion. We next investigated the immunomodulatory impact of VCG, in comparison with the more established Th1-promoting adjuvants, CpG and FL, on rPmp18.1-mediated innate immune activation. Finally, the effect of siRNA targeting TLR4, MyD88, NF-κB p50, and Caspase-1 mRNA in DCs on IL-1β cytokine secretion was also investigated. Bone marrow-derived dendritic cells (BMDCs) were stimulated with rPmp18.1 in the presence or absence of VCG or CpG or FL and the magnitude of cytokines produced was assessed using a multiplex cytokine ELISA assay. Expression of costimulatory molecules and Toll-like receptors (TLRs) was analyzed by flow cytometry. Quantitation of intracellular levels of myeloid differentiation factor 88 (MyD88), nuclear factor kappa beta (NF-κB p50/p65), and Caspase-1 was evaluated by Western immunoblotting analysis while NF-κB p65 nuclear translocation was assessed by confocal microscopy. The results showed DC stimulation with rPmp18.1 provoked the secretion of proinflammatory cytokines and upregulated expression of TLRs and co-stimulatory molecules associated with DC maturation. These responses were significantly (p ≤ 0.001) enhanced by VCG but not CpG or FL. In addition, rPmp18.1 activated the expression of MyD88, NF-κB p50, and Caspase-1 as well as the nuclear expression of NF-κB p65 in treated DCs. Furthermore, targeting TLR4, MyD88, NF-κB p50, and Caspase-1 mRNA in BMDCs with siRNA significantly reduced their expression levels, resulting in decreased IL-1β cytokine

  18. Isolation of two biologically active cell surface proteins from Brucella abortus by chromatofocusing

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Tabatabai, L.B.; Deyoe, B.L.

    1983-01-01

    Brucella abortus contains a group of immunogenic cell surface proteins which have potential value as a vaccine or as a diagnostic reagent for the prevention and diagnosis of bovine brucellosis. Under nondenaturing conditions, these proteins range in molecular weight from 10,000-124,000, as determined by high performance liquid chromatography (HPLC) on TSK 3000sw. By analytical isoelectrofocusing, 6 major protein bands could be distinguished with pI's ranging from 4.0 to 6.0 and 3 additional major proteins with pI's of 7.5, 9.5, and 10. By chromatofocusing on Polybuffer Exchanger 94 with a pH gradient from 6-4, two of the six proteins from pImore » 4-6 were separated, a pI 4.9 and a pI 4.7 protein; a third fraction contained the high pI proteins. The former two proteins were homogeneous by analytical isoelectrofocusing, and a molecular weight of 54,000 daltons was found for both protein species by HPLC on TSK 3000sw. The pI 4-6 and not the pI 9.5 and 10 proteins, could be radiolabeled when intact cells were radioiodinated with diazotized (/sup 125/I)-iodosulfanilic acid. Biological activity of the proteins as assessed in lemmings indicated that immunization with the pI 4.7 and 4.9 proteins afforded better protection against experimental brucellosis than immunization with the high pI proteins. These results support our view that a single surface protein may be sufficient for the prevention of experimental brucellosis.« less

  19. Oral Immunization with a Multivalent Epitope-Based Vaccine, Based on NAP, Urease, HSP60, and HpaA, Provides Therapeutic Effect on H. pylori Infection in Mongolian gerbils.

    PubMed

    Guo, Le; Yang, Hua; Tang, Feng; Yin, Runting; Liu, Hongpeng; Gong, Xiaojuan; Wei, Jun; Zhang, Ying; Xu, Guangxian; Liu, Kunmei

    2017-01-01

    Epitope-based vaccine is a promising strategy for therapeutic vaccination against Helicobacter pylori ( H. pylori ) infection. A multivalent subunit vaccine containing various antigens from H. pylori is superior to a univalent subunit vaccine. However, whether a multivalent epitope-based vaccine is superior to a univalent epitope-based vaccine in therapeutic vaccination against H. pylori , remains unclear. In this study, a multivalent epitope-based vaccine named CWAE against H. pylori urease, neutrophil-activating protein (NAP), heat shock protein 60 (HSP60) and H. pylori adhesin A (HpaA) was constructed based on mucosal adjuvant cholera toxin B subunit (CTB), Th1-type adjuvant NAP, multiple copies of selected B and Th cell epitopes (UreA 27-53 , UreA 183-203 , HpaA 132-141 , and HSP60 189-203 ), and also the epitope-rich regions of urease B subunit (UreB 158-251 and UreB 321-385 ) predicted by bioinformatics. Immunological properties of CWAE vaccine were characterized in BALB/c mice model. Its therapeutic effect was evaluated in H. pylori -infected Mongolian gerbil model by comparing with a univalent epitope-based vaccine CTB-UE against H. pylori urease that was constructed in our previous studies. Both CWAE and CTB-UE could induce similar levels of specific antibodies against H. pylori urease, and had similar inhibition effect of H. pylori urease activity. However, only CWAE could induce high levels of specific antibodies to NAP, HSP60, HpaA, and also the synthetic peptides epitopes (UreB 158-172 , UreB 181-195 , UreB 211-225 , UreB 349-363 , HpaA 132-141 , and HSP60 189-203 ). In addition, oral therapeutic immunization with CWAE significantly reduced the number of H. pylori colonies in the stomach of Mongolian gerbils, compared with oral immunization using CTB-UE or H. pylori urease. The protection of CWAE was associated with higher levels of mixed CD4 + T cell (Th cell) response, IgG, and secretory IgA (sIgA) antibodies to H. pylori . These results indic ate

  20. Oral Immunization with a Multivalent Epitope-Based Vaccine, Based on NAP, Urease, HSP60, and HpaA, Provides Therapeutic Effect on H. pylori Infection in Mongolian gerbils

    PubMed Central

    Guo, Le; Yang, Hua; Tang, Feng; Yin, Runting; Liu, Hongpeng; Gong, Xiaojuan; Wei, Jun; Zhang, Ying; Xu, Guangxian; Liu, Kunmei

    2017-01-01

    Epitope-based vaccine is a promising strategy for therapeutic vaccination against Helicobacter pylori (H. pylori) infection. A multivalent subunit vaccine containing various antigens from H. pylori is superior to a univalent subunit vaccine. However, whether a multivalent epitope-based vaccine is superior to a univalent epitope-based vaccine in therapeutic vaccination against H. pylori, remains unclear. In this study, a multivalent epitope-based vaccine named CWAE against H. pylori urease, neutrophil-activating protein (NAP), heat shock protein 60 (HSP60) and H. pylori adhesin A (HpaA) was constructed based on mucosal adjuvant cholera toxin B subunit (CTB), Th1-type adjuvant NAP, multiple copies of selected B and Th cell epitopes (UreA27–53, UreA183–203, HpaA132–141, and HSP60189–203), and also the epitope-rich regions of urease B subunit (UreB158–251 and UreB321–385) predicted by bioinformatics. Immunological properties of CWAE vaccine were characterized in BALB/c mice model. Its therapeutic effect was evaluated in H. pylori-infected Mongolian gerbil model by comparing with a univalent epitope-based vaccine CTB-UE against H. pylori urease that was constructed in our previous studies. Both CWAE and CTB-UE could induce similar levels of specific antibodies against H. pylori urease, and had similar inhibition effect of H. pylori urease activity. However, only CWAE could induce high levels of specific antibodies to NAP, HSP60, HpaA, and also the synthetic peptides epitopes (UreB158–172, UreB181–195, UreB211–225, UreB349–363, HpaA132–141, and HSP60189–203). In addition, oral therapeutic immunization with CWAE significantly reduced the number of H. pylori colonies in the stomach of Mongolian gerbils, compared with oral immunization using CTB-UE or H. pylori urease. The protection of CWAE was associated with higher levels of mixed CD4+ T cell (Th cell) response, IgG, and secretory IgA (sIgA) antibodies to H. pylori. These results indic ate that a

  1. Central Memory CD4+ T Cells Are Responsible for the Recombinant Bacillus Calmette-Guérin ΔureC::hly Vaccine's Superior Protection Against Tuberculosis

    PubMed Central

    Vogelzang, Alexis; Perdomo, Carolina; Zedler, Ulrike; Kuhlmann, Stefanie; Hurwitz, Robert; Gengenbacher, Martin; Kaufmann, Stefan H. E.

    2014-01-01

    Bacillus Calmette-Guérin (BCG) has been used for vaccination against tuberculosis for nearly a century. Here, we analyze immunity induced by a live tuberculosis vaccine candidate, recombinant BCG ΔureC::hly vaccine (rBCG), with proven preclinical and clinical safety and immunogenicity. We pursue in-depth analysis of the endogenous mycobacteria-specific CD4+ T-cell population, comparing the more efficacious rBCG with canonical BCG to determine which T-cell memory responses are prerequisites for superior protection against tuberculosis. rBCG induced higher numbers and proportions of antigen-specific memory CD4+ T cells than BCG, with a CXCR5+CCR7+ phenotype and low expression of the effector transcription factors T-bet and Bcl-6. We found that the superior protection of rBCG, compared with BCG, correlated with higher proportions and numbers of these central memory T cells and of T follicular helper cells associated with specific antibody responses. Adoptive transfer of mycobacteria-specific central memory T cells validated their critical role in protection against pulmonary tuberculosis. PMID:24943726

  2. Pathology of brucellosis in bison from Yellowstone National Park

    USGS Publications Warehouse

    Rhyan, Jack C.; Gidlewski, T.; Roffe, T.J.; Aune, K.; Philo, L.M.; Ewalt, D.R.

    2001-01-01

    Between February 1995 and June 1999, specimens from seven aborted bison (Bison bison) fetuses or stillborn calves and their placentas, two additional placentas, three dead neonates, one 2-wk-old calf, and 35 juvenile and adult female bison from Yellowstone National Park (USA) were submitted for bacteriologic and histopathologic examination. One adult animal with a retained placenta had recently aborted. Serum samples from the 35 juvenile and adult bison were tested for Brucella spp. antibodies. Twenty-six bison, including the cow with the retained placenta, were seropositive, one was suspect, and eight were seronegative. Brucella abortus biovar 1 was isolated from three aborted fetuses and associated placentas, an additional placenta, the 2-wk-old calf, and 11 of the seropositive female bison including the animal that had recently aborted. Brucella abortus biovar 2 was isolated from one additional seropositive adult female bison. Brucella abortus was recovered from numerous tissue sites from the aborted fetuses, placentas and 2-wk-old calf. In the juvenile and adult bison, the organism was more frequently isolated from supramammary (83%), retropharyngeal (67%), and iliac (58%) lymph nodes than from other tissues cultured. Cultures from the seronegative and suspect bison were negative for B. abortus. Lesions in the B. abortus-infected, aborted placentas and fetuses consisted of necropurulent placentitis and mild bronchointerstitial pneumonia. The infected 2-wk-old calf had bronchointerstitial pneumonia, focal splenic infarction, and purulent nephritis. The recently-aborting bison cow had purulent endometritis and necropurulent placentitis. Immunohistochemical staining of tissues from the culture-positive aborted fetuses, placentas, 2-wk-old calf, and recently-aborting cow disclosed large numbers of B. abortus in placental trophoblasts and exudate, and fetal and calf lung. A similar study with the same tissue collection and culture protocol was done using six

  3. Identification and characterization of proteins involved in rice urea and arginine catabolism.

    PubMed

    Cao, Feng-Qiu; Werner, Andrea K; Dahncke, Kathleen; Romeis, Tina; Liu, Lai-Hua; Witte, Claus-Peter

    2010-09-01

    Rice (Oryza sativa) production relies strongly on nitrogen (N) fertilization with urea, but the proteins involved in rice urea metabolism have not yet been characterized. Coding sequences for rice arginase, urease, and the urease accessory proteins D (UreD), F (UreF), and G (UreG) involved in urease activation were identified and cloned. The functionality of urease and the urease accessory proteins was demonstrated by complementing corresponding Arabidopsis (Arabidopsis thaliana) mutants and by multiple transient coexpression of the rice proteins in Nicotiana benthamiana. Secondary structure models of rice (plant) UreD and UreF proteins revealed a possible functional conservation to bacterial orthologs, especially for UreF. Using amino-terminally StrepII-tagged urease accessory proteins, an interaction between rice UreD and urease could be shown. Prokaryotic and eukaryotic urease activation complexes seem conserved despite limited protein sequence conservation for UreF and UreD. In plant metabolism, urea is generated by the arginase reaction. Rice arginase was transiently expressed as a carboxyl-terminally StrepII-tagged fusion protein in N. benthamiana, purified, and biochemically characterized (K(m) = 67 mm, k(cat) = 490 s(-1)). The activity depended on the presence of manganese (K(d) = 1.3 microm). In physiological experiments, urease and arginase activities were not influenced by the external N source, but sole urea nutrition imbalanced the plant amino acid profile, leading to the accumulation of asparagine and glutamine in the roots. Our data indicate that reduced plant performance with urea as N source is not a direct result of insufficient urea metabolism but may in part be caused by an imbalance of N distribution.

  4. [Seroprevalance of brucellosis, listeriosis and toxoplasmosis in cattle in Adana province of Turkey].

    PubMed

    Yağcı Yücel, Sükran; Yaman, Mehmet; Kurt, Cemal; Babür, Cahit; Celebi, Bekir; Kılıç, Selçuk; Ozen, Doğukan

    2014-06-01

    This study was conducted to identify the seroprevalance of diseases which are zoonotic and responsible from abortion such as toxoplasmosis, listeriosis and brucellosis in Holstein crossbred cattle of different age and sex in Adana province, between 2008 April-September. For this purpose, blood serum samples were collected from 132 cattle and analyzed for Toxoplasma gondii, Listeria monocytogenes and Brucella abortus antibodies. T. gondii, L. monocytogenes and Brucella abortus antibodies were determined by the standard Sabin- Feldman Dye Test (SFDT), Osebold method and Microtube Agglutination Test (MAT) respectively, from the blood serum samples. 132 serum tested 56.06% samples of T. gondii, 40.9% and 3.03% of L. monocytogenes and defined the B.abortus antibodies were found to be seropositive terms. There were no statistically significant difference between seropositive T. gondii, L. monocytogenes and B.abortus antibodies among age groups (p>0.05). In this study, for the first time in cattle in the region of Adana serological methods revealed the presence L. monocytogenes, toxoplasmosis and listeriosis were higher than brucellosis seropositivity. Moreover, the prevalence of these diseases in the same animal at the highest rate was determined for T. gondii and L. monocytogenes.

  5. Comparison of potential protection conferred by three immunization strategies (protein/protein, DNA/DNA, and DNA/protein) against Brucella infection using Omp2b in BALB/c Mice.

    PubMed

    Golshani, Maryam; Rafati, Sima; Nejati-Moheimani, Mehdi; Ghasemian, Melina; Bouzari, Saeid

    2016-12-25

    In the present study, immunogenicity and protective efficacy of the Brucella outer membrane protein 2b (Omp2b) was evaluated in BALB/c mice using Protein/Protein, DNA/DNA and DNA/Protein vaccine strategies. Immunization of mice with three vaccine regimens elicited a strong specific IgG response (higher IgG2a titers over IgG1 titers) and provided Th1-oriented immune response. Vaccination of BALB/c mice with the DNA/Pro regimen induced higher levels of IFN-γ/IL-2 and conferred more protection levels against B. melitenisis and B. abortus challenge than did the protein or DNA alone. In conclusion, Omp2b is able to stimulate specific immune responses and to confer cross protection against B. melitensis and B. abortus infection. Therefore, it could be introduced as a new potential candidate for the development of a subunit vaccine against Brucella infection. Copyright © 2016 Elsevier B.V. All rights reserved.

  6. Altered frontal inter-hemispheric resting state functional connectivity is associated with bulimic symptoms among restrained eaters.

    PubMed

    Chen, Shuaiyu; Dong, Debo; Jackson, Todd; Su, Yanhua; Chen, Hong

    2016-01-29

    Theory and research have indicated that restrained eating (RE) increases risk for binge-eating and eating disorder symptoms. According to the goal conflict model, such risk may result from disrupted hedonic-feeding control and its interaction with reward-driven eating. However, RE-related alterations in functional interactions among associated underlying brain regions, especially between the cerebral hemispheres, have rarely been examined directly. Therefore, we investigated inter-hemispheric resting-state functional connectivity (RSFC) among female restrained eaters (REs) (n=23) and unrestrained eaters (UREs) (n=24) following food deprivation as well as its relation to overall bulimia nervosa (BN) symptoms using voxel-mirrored homotopic connectivity (VMHC). Seed-based RSFC associated with areas exhibiting significant VMHC differences was also assessed. Compared to UREs, REs showed reduced VMHC in the dorsal-lateral prefrontal cortex (DLPFC), an area involved in inhibiting hedonic overeating. REs also displayed decreased RSFC between the right DLPFC and regions associated with reward estimation--the ventromedial prefrontal cortex (VMPFC) and posterior cingulate cortex (PCC). Finally, bulimic tendencies had a negative correlation with VMHC in the DLPFC and a positive correlation with functional connectivity (DLPFC and VMPFC) among REs but not UREs. Findings suggested that reduced inter-hemispheric functional connectivity in appetite inhibition regions and altered functional connectivity in reward related regions may help to explain why some REs fail to control hedonically-motivated feeding and experience higher associated levels of BN symptomatology. Copyright © 2015 Elsevier Ltd. All rights reserved.

  7. Biosynthesis of the Urease Metallocenter*

    PubMed Central

    Farrugia, Mark A.; Macomber, Lee; Hausinger, Robert P.

    2013-01-01

    Metalloenzymes often require elaborate metallocenter assembly systems to create functional active sites. The medically important dinuclear nickel enzyme urease provides an excellent model for studying metallocenter assembly. Nickel is inserted into the urease active site in a GTP-dependent process with the assistance of UreD/UreH, UreE, UreF, and UreG. These accessory proteins orchestrate apoprotein activation by delivering the appropriate metal, facilitating protein conformational changes, and possibly providing a requisite post-translational modification. The activation mechanism and roles of each accessory protein in urease maturation are the subject of ongoing studies, with the latest findings presented in this minireview. PMID:23539618

  8. Translational errors as an early event in prion conversion.

    PubMed

    Hatin, I; Bidou, L; Cullin, C; Rousset, J P

    2001-01-01

    A prion is an infectious, altered form of a cellular protein which can self-propagate and affect normal phenotype. Prion conversion has been observed for mammalian and yeast proteins but molecular mechanisms that trigger this process remain unclear. Up to now, only post-translational models have been explored. In this work, we tested the hypothesis that co-translational events may be implicated in the conformation changes of the Ure2p protein of Saccharomyces cerevisiae. This protein can adopt a prion conformation leading to an [URE3] phenotype which can be easily assessed and quantified. We analyzed the effect of two antibiotics, known to affect translation, on [URE3] conversion frequency. For cells treated with G418 we observed a parallel increase of translational errors rate and frequency of [URE3] conversion. By contrast, cycloheximide which was not found to affect translational fidelity, has no influence on the induction of [URE3] phenotype. These results raise the possibility that the mechanism of prion conversion might not only involve alternative structures of strictly identical molecules but also aberrant proteins resulting from translational errors.

  9. Influence of uncorrected refractive error and unmet refractive error on visual impairment in a Brazilian population

    PubMed Central

    2014-01-01

    Background The World Health Organization (WHO) definitions of blindness and visual impairment are widely based on best-corrected visual acuity excluding uncorrected refractive errors (URE) as a visual impairment cause. Recently, URE was included as a cause of visual impairment, thus emphasizing the burden of visual impairment due to refractive error (RE) worldwide is substantially higher. The purpose of the present study is to determine the reversal of visual impairment and blindness in the population correcting RE and possible associations between RE and individual characteristics. Methods A cross-sectional study was conducted in nine counties of the western region of state of São Paulo, using systematic and random sampling of households between March 2004 and July 2005. Individuals aged more than 1 year old were included and were evaluated for demographic data, eye complaints, history, and eye exam, including no corrected visual acuity (NCVA), best corrected vision acuity (BCVA), automatic and manual refractive examination. The definition adopted for URE was applied to individuals with NCVA > 0.15 logMAR and BCVA ≤ 0.15 logMAR after refractive correction and unmet refractive error (UREN), individuals who had visual impairment or blindness (NCVA > 0.5 logMAR) and BCVA ≤ 0.5 logMAR after optical correction. Results A total of 70.2% of subjects had normal NCVA. URE was detected in 13.8%. Prevalence of 4.6% of optically reversible low vision and 1.8% of blindness reversible by optical correction were found. UREN was detected in 6.5% of individuals, more frequently observed in women over the age of 50 and in higher RE carriers. Visual impairment related to eye diseases is not reversible with spectacles. Using multivariate analysis, associations between URE and UREN with regard to sex, age and RE was observed. Conclusion RE is an important cause of reversible blindness and low vision in the Brazilian population. PMID:24965318

  10. Comparison of DMSP and SECS region-1 and region-2 ionospheric current boundary

    NASA Astrophysics Data System (ADS)

    Weygand, J. M.; Wing, S.

    2016-06-01

    The region-1 and region-2 boundary has traditionally been identified using data from a single spacecraft crossing the auroral region and measuring the large scale changes in the cross track magnetic field. With data from the AUTUMN, CANMOS, CARISMA, GIMA, DTU MGS, MACCS, McMAC, STEP, THEMIS, and USGS ground magnetometer arrays we applied a state-of-art technique based on spherical elementary current system (SECS) method developed by Amm and Viljanen (1999) in order to calculate maps of region-1 and region-2 current system over the North American and Greenland auroral region. Spherical elementary current (SEC) amplitude (proxy for vertical currents) maps can be inferred at 10 s temporal resolution, ~1.5° geographic latitude (Glat), and 3.5° geographic longitude (Glon) spatial resolution. We compare the location of the region-1 and region-2 boundary obtained by the DMSP spacecraft with the region-1 and region-2 boundary observed in the SEC current amplitudes. We find that the boundaries typically agree within 0.2°±1.3°. These results indicate that the location of the region-1 and region-2 boundary can reasonably be determined from ground magnetometer data. The SECS maps represent a value-added product from the magnetometer database and can be used for contextual interpretation in conjunction with other missions as well as help with our understanding of magnetosphere-ionosphere coupling mechanisms using the ground arrays and the magnetospheric spacecraft data.

  11. Comparison of DMSP and SECS region-1 and region-2 ionospheric current boundary✩

    PubMed Central

    Weygand, J.M.; Wing, S.

    2017-01-01

    The region-1 and region-2 boundary has traditionally been identified using data from a single spacecraft crossing the auroral region and measuring the large scale changes in the cross track magnetic field. With data from the AUTUMN, CANMOS, CARISMA, GIMA, DTU MGS, MACCS, McMAC, STEP, THEMIS, and USGS ground magnetometer arrays we applied a state-of-art technique based on spherical elementary current system (SECS) method developed by Amm and Viljanen (1999) in order to calculate maps of region-1 and region-2 current system over the North American and Greenland auroral region. Spherical elementary current (SEC) amplitude (proxy for vertical currents) maps can be inferred at 10 s temporal resolution, ~1.5° geographic latitude (Glat), and 3.5° geographic longitude (Glon) spatial resolution. We compare the location of the region-1 and region-2 boundary obtained by the DMSP spacecraft with the region-1 and region-2 boundary observed in the SEC current amplitudes. We find that the boundaries typically agree within 0.2° ± 1.3°. These results indicate that the location of the region-1 and region-2 boundary can reasonably be determined from ground magnetometer data. The SECS maps represent a value-added product from the magnetometer database and can be used for contextual interpretation in conjunction with other missions as well as help with our understanding of magnetosphere-ionosphere coupling mechanisms using the ground arrays and the magnetospheric spacecraft data. PMID:29056861

  12. Comparison of DMSP and SECS region-1 and region-2 ionospheric current boundary.

    PubMed

    Weygand, J M; Wing, S

    2016-06-01

    The region-1 and region-2 boundary has traditionally been identified using data from a single spacecraft crossing the auroral region and measuring the large scale changes in the cross track magnetic field. With data from the AUTUMN, CANMOS, CARISMA, GIMA, DTU MGS, MACCS, McMAC, STEP, THEMIS, and USGS ground magnetometer arrays we applied a state-of-art technique based on spherical elementary current system (SECS) method developed by Amm and Viljanen (1999) in order to calculate maps of region-1 and region-2 current system over the North American and Greenland auroral region. Spherical elementary current (SEC) amplitude (proxy for vertical currents) maps can be inferred at 10 s temporal resolution, ~1.5° geographic latitude (Glat), and 3.5° geographic longitude (Glon) spatial resolution. We compare the location of the region-1 and region-2 boundary obtained by the DMSP spacecraft with the region-1 and region-2 boundary observed in the SEC current amplitudes. We find that the boundaries typically agree within 0.2° ± 1.3°. These results indicate that the location of the region-1 and region-2 boundary can reasonably be determined from ground magnetometer data. The SECS maps represent a value-added product from the magnetometer database and can be used for contextual interpretation in conjunction with other missions as well as help with our understanding of magnetosphere-ionosphere coupling mechanisms using the ground arrays and the magnetospheric spacecraft data.

  13. 50 CFR 2.1 - Regional offices.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 50 Wildlife and Fisheries 1 2010-10-01 2010-10-01 false Regional offices. 2.1 Section 2.1 Wildlife... FIELD ORGANIZATION § 2.1 Regional offices. The program operations of the U.S. Fish and Wildlife Service..., endangered species stations, fishery assistance offices, national fish hatcheries, national wildlife refuges...

  14. Effects of the hook of Uncaria rhynchophylla on neurotoxicity in the 6-hydroxydopamine model of Parkinson's disease.

    PubMed

    Shim, Jin Sup; Kim, Hyo Geun; Ju, Mi Sun; Choi, Jin Gyu; Jeong, Seo Young; Oh, Myung Sook

    2009-11-12

    While the hook of Uncaria rhynchophylla (URH) is a traditional herb used in northeast Asia for the treatment of Parkinson's disease (PD)-like symptoms such as tremor, it has not been experimentally evaluated in a PD model. We investigated the effects of URH on 6-hydroxydapamine (6-OHDA)-induced neurotoxicity in in vitro and in vivo models of PD. The cell viability, anti-oxidative activity, and anti-apoptotic activity of a water extract of URH (URE) were assessed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide, reactive oxygen species (ROS), total glutathione (GSH), and caspase-3 assays in PC12 cells stressed by 6-OHDA. We also investigated the behavioral recovery and dopaminergic neuron protection of URE using an apomorphine-induced rotation test and tyrosine hydroxylase immunohistochemistry in the hemi-parkinsonian rat model of the unilateral 6-OHDA lesion of the medial forebrain bundle. In PC12 cells, URE significantly reduced cell death and the generation of ROS, increased GSH levels, and inhibited caspase-3 activity induced by 6-OHDA. In 6-OHDA-lesioned rats, posttreatment with URE (5 mg/kg/day for 14 days) significantly reduced apomorphine-induced rotation, and it lowered dopaminergic neuronal loss in substantia nigra pars compacta. URE possesses neuroprotective activity against 6-OHDA-induced toxicity through anti-oxidative and anti-apoptotic activities in PD models.

  15. Pathogenic outcome following experimental infection of sheep with Chlamydia abortus variant strains LLG and POS.

    PubMed

    Livingstone, Morag; Wheelhouse, Nicholas; Ensor, Hannah; Rocchi, Mara; Maley, Stephen; Aitchison, Kevin; Wattegedera, Sean; Wilson, Kim; Sait, Michelle; Siarkou, Victoria; Vretou, Evangelia; Entrican, Gary; Dagleish, Mark; Longbottom, David

    2017-01-01

    This study investigated the pathogenesis of two variant strains (LLG and POS) of Chlamydia abortus, in comparison to a typical wild-type strain (S26/3) which is known to be responsible for late term abortion in small ruminants. Challenge with the three strains at mid-gestation resulted in similar pregnancy outcomes, with abortion occurring in approximately 50-60% of ewes with the mean gestational lengths also being similar. However, differences were observed in the severity of placental pathology, with infection appearing milder for strain LLG, which was reflected in the lower number of organisms shed in vaginal swabs post-partum and less gross pathology and organisms present in placental smears. Results for strain POS were somewhat different than LLG with a more focal restriction of infection observed. Post-abortion antibody responses revealed prominent differences in seropositivity to the major outer membrane protein (MOMP) present in elementary body (EB) preparations under denaturing conditions, most notably with anti-LLG and anti-POS convalescent sera where there was no or reduced detection of MOMP present in EBs derived from the three strains. These results and additional analysis of whole EB and chlamydial outer membrane complex preparations suggest that there are conformational differences in MOMP for the three strains. Overall, the results suggest that gross placental pathology and clinical outcome is not indicative of bacterial colonization and the severity of infection. The results also highlight potential conformational differences in MOMP epitopes that perhaps impact on disease diagnosis and the development of new vaccines.

  16. T cell regulation of the thymus-independent antibody response to trinitrophenylated-Brucella abortus (TNP-BA)

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Tanay, A.; Strober, S.

    The authors have previously observed a reduction of the T cell-dependent primary antibody response to dinitrophenylated keyhole limpet hemocyanin, and an enhancement of the T cell-independent response to trinitrophenylated Brucella abortus (TNP-BA) in BALB/c mice after treatment with total lymphoid irradiation (TLI). To elucidate the relative contribution of T and B cells to the enhanced T cell-independent antibody responses after TLI, a syngeneic primary adoptive transfer system was utilized whereby irradiated hosts were reconstituted with unfractionated spleen cells or a combination of purified T and B cells from TLI-treated and untreated control mice. Antibody responses of purified splenic B cellsmore » from TLI-treated BALB/c mice (TLI/B) to TNP-BA were enhanced 10-fold as compared with those of unfractionated (UF) spleen cells or B cells from normal (NL) BALB/c mice (NL/UF and NL/B, respectively). Splenic T cells from normal animals (NL/T) suppressed the anti-TNP-BA response of TLI/B by more than 100-fold. NL/T neither suppressed nor enhanced the response of NL/B. On the other hand, T cells from TLI-treated mice (TLI/T) enhanced by 100-fold the anti-TNP-BA response of NL/B, but neither suppressed nor enhanced the response of TLI/B. Thus, T cells can regulate the T cell-independent antibody response to TNP-BA. However, experimental manipulation of the T and B cell populations is needed to demonstrate the regulatory functions.« less

  17. The Reactions of Nitrogen Peroxide with Possible Stabilisers for Propellants

    DTIC Science & Technology

    1957-03-01

    ether Carbamite Phe nyl-be nzyl-ure thane (pure) Cyclohexanyl-urethane Cyclohexano ne Die thyl phthalate Di-isoamyl phthalate Dibutyl oxalate Glycollic...saponification" arises from the presence of phenyl urethane and diphenyl urea; differences in contents of these impurities and of benzyl aniline...nitrogen that is recovered from a product. 4.2.2 Ure are fairly reactive. Triphenylethylurea present with diphenyl - amine in 蠢 compound" leads to a

  18. Ureolytic Biomineralization Reduces Proteus mirabilis Biofilm Susceptibility to Ciprofloxacin

    PubMed Central

    Li, Xiaobao; Lu, Nanxi; Brady, Hannah R.

    2016-01-01

    Ureolytic biomineralization induced by urease-producing bacteria, particularly Proteus mirabilis, is responsible for the formation of urinary tract calculi and the encrustation of indwelling urinary catheters. Such microbial biofilms are challenging to eradicate and contribute to the persistence of catheter-associated urinary tract infections, but the mechanisms responsible for this recalcitrance remain obscure. In this study, we characterized the susceptibility of wild-type (ure+) and urease-negative (ure−) P. mirabilis biofilms to killing by ciprofloxacin. Ure+ biofilms produced fine biomineral precipitates that were homogeneously distributed within the biofilm biomass in artificial urine, while ure− biofilms did not produce biomineral deposits under identical growth conditions. Following exposure to ciprofloxacin, ure+ biofilms showed greater survival (less killing) than ure− biofilms, indicating that biomineralization protected biofilm-resident cells against the antimicrobial. To evaluate the mechanism responsible for this recalcitrance, we observed and quantified the transport of Cy5-conjugated ciprofloxacin into the biofilm by video confocal microscopy. These observations revealed that the reduced susceptibility of ure+ biofilms resulted from hindered delivery of ciprofloxacin into biomineralized regions of the biofilm. Further, biomineralization enhanced retention of viable cells on the surface following antimicrobial exposure. These findings together show that ureolytic biomineralization induced by P. mirabilis metabolism strongly regulates antimicrobial susceptibility by reducing internal solute transport and increasing biofilm stability. PMID:26953206

  19. Comparing the Impact of Refractive and Non-Refractive Vision Loss on Functioning and Disability: The Salisbury Eye Evaluation

    PubMed Central

    Zebardast, Nazlee; Swenor, Bonnielin K.; van Landingham, Suzanne W.; Massof, Robert W.; Munoz, Beatriz; West, Sheila K.; Ramulu, Pradeep Y.

    2015-01-01

    Purpose To compare the effects of uncorrected refractive error (URE) and non-refractive visual impairment (VI) on performance and disability measures. Design Cross-sectional population-based study. Participants 2469 individuals with binocular presenting visual acuity (PVA) of 20/80 or better who participated in the first round of the Salisbury Eye Evaluation study. Methods URE was defined as binocular PVA of 20/30 or worse, improving to better than 20/30 with subjective refraction. VI was defined as post-refraction binocular best corrected visual acuity (BCVA) of 20/30 or worse. The visual acuity decrement attributable to VI was calculated as the difference between BCVA and 20/30 while that due to URE was taken as the difference between PVA and BCVA. Multivariable regression analyses were used to assess the disability impact of 1) vision status (VI, URE, or normal vision) using the group with normal vision as reference, and 2) a one-line decrement in acuity due to VI or URE. Main Outcome Measures Objective measures of visual function were obtained from timed performance of mobility and near vision tasks, self-reported driving cessation, and self-reported visual difficulty measured by the Activities of Daily Vision (ADV) scale. ADV responses were analyzed using Rasch analysis to determine visual ability. Results Compared to individuals with normal vision, subjects with VI (n=191) had significantly poorer objective and subjective visual functioning in all metrics examined (p<0.05) while subjects with URE (n=132) demonstrated slower walking speeds, slower near task performance, more frequent driving cessation and lower ADV scores (p<0.05), but did not demonstrate slower stair climbing or descent speed. For all functional metrics evaluated, the impact of VI was greater than the impact of URE. The impact of a one-line VA decrement due to VI was associated with greater deficits in mobility measures and driving cessation when compared to a one-line VA decrement due to URE

  20. Comparing the Impact of Refractive and Nonrefractive Vision Loss on Functioning and Disability: The Salisbury Eye Evaluation.

    PubMed

    Zebardast, Nazlee; Swenor, Bonnielin K; van Landingham, Suzanne W; Massof, Robert W; Munoz, Beatriz; West, Sheila K; Ramulu, Pradeep Y

    2015-06-01

    To compare the effects of uncorrected refractive error (URE) and nonrefractive visual impairment (VI) on performance and disability measures. Cross-sectional, population-based study. A total of 2469 individuals with binocular presenting visual acuity (PVA) of ≥ 20/80 who participated in the first round of the Salisbury Eye Evaluation study. The URE was defined as binocular PVA of ≤ 20/30, improving to >20/30 with subjective refraction. The VI was defined as post-refraction binocular best-corrected visual acuity (BCVA) of ≤ 20/30. The visual acuity decrement due to VI was calculated as the difference between BCVA and 20/30, whereas visual acuity due to URE was taken as the difference between PVA and BCVA. Multivariable regression analyses were used to assess the disability impact of (1) vision status (VI, URE, or normal vision) using the group with normal vision as reference and (2) a 1-line decrement in acuity due to VI or URE. Objective measures of visual function were obtained from timed performance of mobility and near vision tasks, self-reported driving cessation, and self-reported visual difficulty measured by the Activities of Daily Vision (ADV) scale. The ADV responses were analyzed using Rasch analysis to determine visual ability. Compared with individuals with normal vision, subjects with VI (n = 191) had significantly poorer objective and subjective visual functioning in all metrics examined (P < 0.05), whereas subjects with URE (n = 132) demonstrated slower walking speeds, slower near task performance, more frequent driving cessation, and lower ADV scores (P < 0.05), but did not demonstrate slower stair climbing or descent speed. For all functional metrics evaluated, the impact of VI was greater than the impact of URE. The impact of a 1-line VA decrement due to VI was associated with greater deficits in mobility measures and driving cessation when compared with a 1-line VA decrement due to URE. Visual impairment is associated with greater

  1. Pancreatic islet-like clusters from bone marrow mesenchymal stem cells of human first-trimester abortus can cure streptozocin-induced mouse diabetes.

    PubMed

    Zhang, Yihua; Shen, Wenzheng; Hua, Jinlian; Lei, Anmin; Lv, Changrong; Wang, Huayan; Yang, Chunrong; Gao, Zhimin; Dou, Zhongying

    2010-12-01

    Bone marrow mesenchymal stem cells (BMSCs) have been reported to possess low immunogenicity and cause immunosuppression of recipients when allografted. They can differentiate into insulin-producing cells and may be a valuable source for islet formation. However, the extremely low differentiating rate of adult BMSCs toward insulin-producing cells and the insufficient insulin secretion of the differentiated BMSCs in vitro prevent their clinical use in diabetes treatment. Little is known about the potential of cell replacement therapy with human BMSCs. Previously, we isolated and identified human first-trimester fetal BMSCs (hfBMSCs). Under a novel four-step induction procedure established in this study, the hfBMSCs effectively differentiated into functional pancreatic islet-like cell clusters that contained 62 ± 14% insulin-producing cells, expressed a broad gene profile related to pancreatic islet β-cell development, and released high levels of insulin (2.245 ± 0.222 pmol/100 clusters per 30 min) and C-peptide (2.200 ± 0.468 pmol/100 clusters per 30 min) in response to 25 mmol/L glucose stimulus in vitro. The pancreatic islet-like cell clusters normalized the blood glucose level of diabetic model mice for at least 9 weeks when xenografted; blood glucose levels in these mice rose abnormally again when the grafts were removed. Examination of the grafts indicated that the transplanted cells survived in recipients and produced human insulin and C-peptide in situ. These results demonstrate that hfBMSCs derived from a human first-trimester abortus can differentiate into pancreatic islet-like cell clusters following an established four-step induction. The insulin-producing clusters present advantages in cell replacement therapy of type 1 diabetic model mice.

  2. Immunologic responses of bison to vaccination with Brucella abortus strain RB51: comparison of parenteral to ballistic delivery via compressed pellets or photopolymerized hydrogels.

    PubMed

    Olsen, Steven C; Christie, R J; Grainger, D W; Stoffregen, W S

    2006-02-27

    This study compared responses of bison calves to 10(10)CFU of Brucella abortus strain RB51 (SRB51) delivered by parenteral or ballistic methods. Two types of biobullet payloads were evaluated; compacted SRB51 pellets or SRB51 encapsulated in photopolymerized poly(ethylene glycol) hydrogels. Bison were vaccinated with saline, parenteral SRB51 alone, or in combination with Spirovac, or ballistically with compressed SRB51 or hydrogel biobullets. Bison parenterally vaccinated with SRB51 had greater (P<0.05) immunologic responses when compared to control bison. Co-administration of Spirovac as an adjuvant did not influence immunologic responses. As compared to compressed SRB51 biobullets, ballistic vaccination with hydrogel biobullets increased cellular immune responses at some sampling times. Our data suggest that hydrogel formulations of SRB51 may be a superior alternative to compressed SRB51 tablets for ballistic vaccination of bison. Although preliminary, data suggests that immunologic responses of bison to SRB51 hydrogel bullets are similar to responses after parenteral vaccination with SRB51.

  3. Assessment of genetic diversity of zoonotic Brucella spp. recovered from livestock in Egypt using multiple locus VNTR analysis.

    PubMed

    Menshawy, Ahmed M S; Perez-Sancho, Marta; Garcia-Seco, Teresa; Hosein, Hosein I; García, Nerea; Martinez, Irene; Sayour, Ashraf E; Goyache, Joaquín; Azzam, Ragab A A; Dominguez, Lucas; Alvarez, Julio

    2014-01-01

    Brucellosis is endemic in most parts of Egypt, where it is caused mainly by Brucella melitensis biovar 3, and affects cattle and small ruminants in spite of ongoing efforts devoted to its control. Knowledge of the predominant Brucella species/strains circulating in a region is a prerequisite of a brucellosis control strategy. For this reason a study aiming at the evaluation of the phenotypic and genetic heterogeneity of a panel of 17 Brucella spp. isolates recovered from domestic ruminants (cattle, buffalo, sheep, and goat) from four governorates during a period of five years (2002-2007) was carried out using microbiological tests and molecular biology techniques (PCR, MLVA-15, and sequencing). Thirteen strains were identified as B. melitensis biovar 3 while all phenotypic and genetic techniques classified the remaining isolates as B. abortus (n = 2) and B. suis biovar 1 (n = 2). MLVA-15 yielded a high discriminatory power (h = 0.801), indicating a high genetic diversity among the B. melitensis strains circulating among domestic ruminants in Egypt. This is the first report of the isolation of B. suis from cattle in Egypt which, coupled with the finding of B. abortus, suggests a potential role of livestock as reservoirs of several zoonotic Brucella species in the region.

  4. INL Researchers Advance Detection of Brucellosis

    ScienceCinema

    Roberto, Frank; Newby, Deborah

    2017-12-09

    What do cattle ranchers in the greater Yellowstone region have in common with British soldiers garrisoned on the island of Malta in the late 1800s? Hint: it's a pathogen that starts with the letter B. It's Brucella Abortus.

  5. INL Researchers Advance Detection of Brucellosis

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Roberto, Frank; Newby, Deborah

    What do cattle ranchers in the greater Yellowstone region have in common with British soldiers garrisoned on the island of Malta in the late 1800s? Hint: it's a pathogen that starts with the letter B. It's Brucella Abortus.

  6. 18 CFR 2.21 - Regional Transmission Groups.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... Groups. 2.21 Section 2.21 Conservation of Power and Water Resources FEDERAL ENERGY REGULATORY COMMISSION... Interpretations Under the Federal Power Act § 2.21 Regional Transmission Groups. (a) General policy. The Commission encourages Regional Transmission Groups (RTGs) as a means of enabling the market for electric...

  7. Silencing urease: a key evolutionary step that facilitated the adaptation of Yersinia pestis to the flea-borne transmission route.

    PubMed

    Chouikha, Iman; Hinnebusch, B Joseph

    2014-12-30

    The arthropod-borne transmission route of Yersinia pestis, the bacterial agent of plague, is a recent evolutionary adaptation. Yersinia pseudotuberculosis, the closely related food-and water-borne enteric species from which Y. pestis diverged less than 6,400 y ago, exhibits significant oral toxicity to the flea vectors of plague, whereas Y. pestis does not. In this study, we identify the Yersinia urease enzyme as the responsible oral toxin. All Y. pestis strains, including those phylogenetically closest to the Y. pseudotuberculosis progenitor, contain a mutated ureD allele that eliminated urease activity. Restoration of a functional ureD was sufficient to make Y. pestis orally toxic to fleas. Conversely, deletion of the urease operon in Y. pseudotuberculosis rendered it nontoxic. Enzymatic activity was required for toxicity. Because urease-related mortality eliminates 30-40% of infective flea vectors, ureD mutation early in the evolution of Y. pestis was likely subject to strong positive selection because it significantly increased transmission potential.

  8. Silencing urease: A key evolutionary step that facilitated the adaptation of Yersinia pestis to the flea-borne transmission route

    PubMed Central

    Chouikha, Iman; Hinnebusch, B. Joseph

    2014-01-01

    The arthropod-borne transmission route of Yersinia pestis, the bacterial agent of plague, is a recent evolutionary adaptation. Yersinia pseudotuberculosis, the closely related food-and water-borne enteric species from which Y. pestis diverged less than 6,400 y ago, exhibits significant oral toxicity to the flea vectors of plague, whereas Y. pestis does not. In this study, we identify the Yersinia urease enzyme as the responsible oral toxin. All Y. pestis strains, including those phylogenetically closest to the Y. pseudotuberculosis progenitor, contain a mutated ureD allele that eliminated urease activity. Restoration of a functional ureD was sufficient to make Y. pestis orally toxic to fleas. Conversely, deletion of the urease operon in Y. pseudotuberculosis rendered it nontoxic. Enzymatic activity was required for toxicity. Because urease-related mortality eliminates 30–40% of infective flea vectors, ureD mutation early in the evolution of Y. pestis was likely subject to strong positive selection because it significantly increased transmission potential. PMID:25453069

  9. Uncaria rhynchophylla Ameliorates Parkinson's Disease by Inhibiting HSP90 Expression: Insights from Quantitative Proteomics.

    PubMed

    Lan, Yu-Long; Zhou, Jun-Jun; Liu, Jing; Huo, Xiao-Kui; Wang, Ya-Li; Liang, Jia-Hao; Zhao, Jian-Chao; Sun, Cheng-Peng; Yu, Zhen-Long; Fang, Lin-Lin; Tian, Xiang-Ge; Feng, Lei; Ning, Jing; Zhang, Bao-Jing; Wang, Chao; Zhao, Xin-Yu; Ma, Xiao-Chi

    2018-06-21

    Uncaria rhynchophylla, known as "Gou-teng", is a traditional Chinese medicine (TCM) used to extinguish wind, clear heat, arrest convulsions, and pacify the liver. Although U. rhynchophylla has a long history of being often used to treat central nervous system (CNS) diseases, its efficacy and potential mechanism are still uncertain. This study investigated neuroprotective effect and the underlying mechanism of U. rhynchophylla extract (URE) in MPP+-induced SH-SY5Y cells and MPTP-induced mice. MPP+-induced SH-SY5Y cells and MPTP-induced mice were used to established Parkinson's disease (PD) models. Quantitative proteomics and bioinformatics were used to uncover proteomics changes of URE. Western blotting was used to validate main differentially expressed proteins and test HSP90 client proteins (apoptosis-related, autophagy-related, MAPKs, PI3K, and AKT proteins). Flow cytometry and JC-1 staining assay were further used to confirm the effect of URE on MPP+-induced apoptosis in SH-SY5Y cells. Gait analysis was used to detect the behavioral changes in MPTP-induced mice. The levels of dopamine (DA) and their metabolites were examined in striatum (STR) by HPLC-EC. The positive expression of tyrosine hydroxylase (TH) was detected by immunohischemical staining and Western blotting. URE dose-dependently increased the cell viability in MPP+-induced SH-SY5Y cells. Quantitative proteomics and bioinformatics results confirmed that HSP90 was an important differentially expressed protein of URE. URE inhibited the expression of HSP90, which further reversed MPP+-induced cell apoptosis and autophagy by increasing the expressions of Bcl-2, Cyclin D1, p-ERK, p-PI3K p85, PI3K p110α, p-AKT, and LC3-I and decreasing cleaved caspase 3, Bax, p-JNK, p-p38, and LC3-II. URE also markedly decreased the apoptotic ratio and elevated mitochondrial transmembrane potential (DΨm). Furthermore, URE treatment ameliorated behavioral impairments, increased the contents of DA and its metabolites and

  10. Brucella suis strain 2 vaccine is safe and protective against heterologous Brucella spp. infections.

    PubMed

    Zhu, Liangquan; Feng, Yu; Zhang, Ge; Jiang, Hui; Zhang, Zhen; Wang, Nan; Ding, Jiabo; Suo, Xun

    2016-01-12

    Brucellosis is a wide spread zoonotic disease that causes abortion and infertility in mammals and leads to debilitating, febrile illness in humans. Brucella abortus, Brucella melitensis and Brucella suis are the major pathogenic species to humans. Vaccination with live attenuated B. suis strain 2 (S2) vaccine is an essential and critical component in the control of brucellosis in China. The S2 vaccine is very effective in preventing brucellosis in goats, sheep, cattle and swine. However, there are still debates outside of China whether the S2 vaccine is able to provide protection against heterologous virulent Brucella species. We investigated the residual virulence, immunogenicity and protective efficacy of the S2 vaccine in BALB/c mice by determining bacteria persistence in spleen, serum antibody response, cellular immune response and protection against a heterologous virulent challenge. The S2 vaccine was of low virulence as there were no bacteria recovered in spleen four weeks post vaccination. The vaccinated mice developed Brucella-specific IgG in 2-3 weeks, and a burst production of IFN-γ at one week as well as a two-fold increase in TNF-α production. The S2 vaccine protected mice from a virulent challenge by B. melitensis M28, B. abortus 2308 and B. suis S1330, and the S2 vaccinated mice did not develop any clinical signs or tissue damage. Our study demonstrated that the S2 vaccine is of low virulence, stimulates good humoral and cellular immunity and protects animals against infection by heterologous, virulent Brucella species. Copyright © 2015 Elsevier Ltd. All rights reserved.

  11. Feasibility of quarantine procedures for bison (Bison bison) calves from Yellowstone National Park for conservation of brucellosis-free bison.

    PubMed

    Ryan Clarke, P; Frey, Rebecca K; Rhyan, Jack C; McCollum, Matt P; Nol, Pauline; Aune, Keith

    2014-03-01

    OBJECTIVE--To determine the feasibility of qualifying individuals or groups of Yellowstone National Park bison as free from brucellosis. DESIGN--Cohort study. SAMPLE--Serum, blood, and various samples from live bison and tissues taken at necropsy from 214 bison over 7 years. PROCEDURES--Blood was collected from bison every 30 to 45 days for serologic tests and microbiological culture of blood for Brucella abortus. Seropositive bison were euthanized until all remaining bison had 2 consecutive negative test results. Half the seronegative bison were randomly euthanized, and tissues were collected for bacteriologic culture. The remaining seronegative bison were bred, and blood was tested at least twice per year. Cow-calf pairs were sampled immediately after calving and 6 months after calving for evidence of B abortus. RESULTS--Post-enrollment serial testing for B abortus antibodies revealed no bison that seroconverted after 205 days (first cohort) and 180 days (second cohort). During initial serial testing, 85% of bison seroconverted within 120 days after removal from the infected population. Brucella abortus was not cultured from any euthanized seronegative bison (0/88). After parturition, no cows or calves had a positive test result for B abortus antibodies, nor was B abortus cultured from any samples. CONCLUSIONS AND CLINICAL RELEVANCE--Results suggested it is feasible to qualify brucellosis-free bison from an infected herd following quarantine procedures as published in the USDA APHIS brucellosis eradication uniform methods and rules. Latent infection was not detected in this sample of bison when applying the USDA APHIS quarantine protocol.

  12. Factors Influencing Student Gains from Undergraduate Research Experiences at a Hispanic-Serving Institution

    PubMed Central

    Daniels, Heather; Grineski, Sara E.; Collins, Timothy W.; Morales, Danielle X.; Morera, Osvaldo; Echegoyen, Lourdes

    2016-01-01

    Undergraduate research experiences (UREs) confer many benefits to students, including improved self-confidence, better communication skills, and an increased likelihood of pursuing science careers. Additionally, UREs may be particularly important for racial/ethnic minority students who are underrepresented in the science workforce. We examined factors hypothetically relevant to underrepresented minority student gains from UREs at a Hispanic-serving institution, such as mentoring quality, family income, being Latino/a, and caring for dependents. Data came from a 2013 survey of University of Texas at El Paso students engaged in 10 URE programs (n = 227). Using generalized linear models (GzLMs) and adjusting for known covariates, we found that students who reported receiving higher-quality mentorship, spending more hours caring for dependents, and receiving more programmatic resources experienced significantly greater gains from their URE in all three areas we examined (i.e., thinking and working like a scientist, personal gains, and gains in skills). In two of three areas, duration of the URE was positive and significant. Being Latino/a was positive and significant only in the model predicting personal gains. Across the three models, quality of mentorship was the most important correlate of gains. This suggests that providing training to faculty mentors involved in UREs may improve student outcomes and increase program efficacy. PMID:27521234

  13. The unexpected discovery of Brucella abortus Buck 19 vaccine in goats from Ecuador underlines the importance of biosecurity measures.

    PubMed

    Ron-Román, Jorge; Berkvens, Dirk; Barzallo-Rivadeneira, Daniela; Angulo-Cruz, Alexandra; González-Andrade, Pablo; Minda-Aluisa, Elizabeth; Benítez-Ortíz, Washington; Brandt, Jef; Rodríguez-Hidalgo, Richar; Saegerman, Claude

    2017-03-01

    Very few, mostly old, and only preliminary serological studies of brucellosis in goats exist in Ecuador. In order to assess the current epidemiological situation, we performed a cross-sectional serological study in the goat populations of Carchi (n = 160 animals), Pichincha (n = 224 animals), and Loja provinces (n = 2024 animals). Only two positive serological results (RB negative and SAT-EDTA ≥400 IU/ml) were obtained in lactating goats from the same farm in Quito (Pichincha province). Additionally, milk was sampled from 220 animals in Pichincha province. The present study indicates a low apparent prevalence in Pichincha province and absence in Carchi and Loja provinces. A total of 25 positive milk ring tests (MRT) were obtained in Pichincha province yielding a prevalence of MRT of 11.16%. Subsequent culture was performed on the positive MRT samples. All results were negative, apart from a single sample, obtained from a serologically positive goat in Quito, that was positive for Brucella abortus strain 19 (B19). Several hypotheses are forwarded concerning this unexpected result. The most likely hypothesis is the possible accidental use of a needle, previously used for vaccination of cattle with the said vaccine, for the administration of drug treatment to the goat. This hypothesis underlines the necessity of biosecurity measures to prevent this type of accidents.

  14. Evolutionary history of the ABCB2 genomic region in teleosts

    USGS Publications Warehouse

    Palti, Y.; Rodriguez, M.F.; Gahr, S.A.; Hansen, J.D.

    2007-01-01

    Gene duplication, silencing and translocation have all been implicated in shaping the unique genomic architecture of the teleost MH regions. Previously, we demonstrated that trout possess five unlinked regions encoding MH genes. One of these regions harbors ABCB2 which in all other vertebrate classes is found in the MHC class II region. In this study, we sequenced a BAC contig for the trout ABCB2 region. Analysis of this region revealed the presence of genes homologous to those located in the human class II (ABCB2, BRD2, ??DAA), extended class II (RGL2, PHF1, SYGP1) and class III (PBX2, Notch-L) regions. The organization and syntenic relationships of this region were then compared to similar regions in humans, Tetraodon and zebrafish to learn more about the evolutionary history of this region. Our analysis indicates that this region was generated during the teleost-specific duplication event while also providing insight about potential MH paralogous regions in teleosts. ?? 2006 Elsevier Ltd. All rights reserved.

  15. Yeast prions assembly and propagation: contributions of the prion and non-prion moieties and the nature of assemblies.

    PubMed

    Kabani, Mehdi; Melki, Ronald

    2011-01-01

    Yeast prions are self-perpetuating protein aggregates that are at the origin of heritable and transmissible non-Mendelian phenotypic traits. Among these, [PSI+], [URE3] and [PIN+] are the most well documented prions and arise from the assembly of Sup35p, Ure2p and Rnq1p, respectively, into insoluble fibrillar assemblies. Fibril assembly depends on the presence of N- or C-terminal prion domains (PrDs) which are not homologous in sequence but share unusual amino-acid compositions, such as enrichment in polar residues (glutamines and asparagines) or the presence of oligopeptide repeats. Purified PrDs form amyloid fibrils that can convert prion-free cells to the prion state upon transformation. Nonetheless, isolated PrDs and full-length prion proteins have different aggregation, structural and infectious properties. In addition, mutations in the "non-prion" domains (non-PrDs) of Sup35p, Ure2p and Rnq1p were shown to affect their prion properties in vitro and in vivo. Despite these evidences, the implication of the functional non-PrDs in fibril assembly and prion propagation has been mostly overlooked. In this review, we discuss the contribution of non-PrDs to prion assemblies, and the structure-function relationship in prion infectivity in the light of recent findings on Sup35p and Ure2p assembly into infectious fibrils from our laboratory and others.

  16. Conformity Adequacy Review: Region 2

    EPA Pesticide Factsheets

    Resources are for air quality and transportation government and community leaders. Information on the adequacy/inadequacy of state implementation plans (SIPs) in EPA Region 2 (NJ, NY, PR, VI) is provided here.

  17. Brucella-Salmonella lipopolysaccharide chimeras are less permeable to hydrophobic probes and more sensitive to cationic peptides and EDTA than are their native Brucella sp. counterparts.

    PubMed Central

    Freer, E; Moreno, E; Moriyón, I; Pizarro-Cerdá, J; Weintraub, A; Gorvel, J P

    1996-01-01

    A rough (R) Brucella abortus 45/20 mutant was more sensitive to the bactericidal activity of polymyxin B and lactoferricin B than was its smooth (S) counterpart but considerably more resistant than Salmonella montevideo. The outer membrane (OM) and isolated lipopolysaccharide (LPS) of S. montevideo showed a higher affinity for these cationic peptides than did the corresponding B. abortus OM and LPS. We took advantage of the moderate sensitivity of R B. abortus to cationic peptides to construct live R B. abortus-S-LPS chimeras to test the activities of polymyxin B, lactoferricin B, and EDTA. Homogeneous and abundant peripheral distribution of the heterologous S-LPS was observed on the surface of the chimeras, and this coating had no effect on the viability or morphology of the cells. When the heterologous LPS corresponded to the less sensitive bacterium S B. abortus S19, the chimeras were more resistant to cationic peptides; in contrast, when the S-LPS was from the more sensitive bacterium S. montevideo, the chimeras were more susceptible to the action of peptides and EDTA. A direct correlation between the amount of heterologous S-LPS on the surface of chimeric Brucella cells and peptide sensitivity was observed. Whereas the damage produced by polymyxin B in S. montevideo and B. abortus-S. montevideo S-LPS chimeras was manifested mainly as OM blebbing and inner membrane rolling, lactoferricin B caused inner membrane detachment, vacuolization, and the formation of internal electron-dense granules in these cells. Native S and R B. abortus strains were permeable to the hydrophobic probe N-phenyl-1-naphthylamine (NPN). In contrast, only reduced amounts of NPN partitioned into the OMs of the S. montevideo and B. abortus-S. montevideo S-LPS chimeras. Following peptide exposure, accelerated NPN uptake similar to that observed for S. montevideo was detected for the B. abortus-S. montevideo LPS chimeras. The partition of NPN into native or EDTA-, polymyxin B-, or

  18. Brucella-Salmonella lipopolysaccharide chimeras are less permeable to hydrophobic probes and more sensitive to cationic peptides and EDTA than are their native Brucella sp. counterparts.

    PubMed

    Freer, E; Moreno, E; Moriyón, I; Pizarro-Cerdá, J; Weintraub, A; Gorvel, J P

    1996-10-01

    A rough (R) Brucella abortus 45/20 mutant was more sensitive to the bactericidal activity of polymyxin B and lactoferricin B than was its smooth (S) counterpart but considerably more resistant than Salmonella montevideo. The outer membrane (OM) and isolated lipopolysaccharide (LPS) of S. montevideo showed a higher affinity for these cationic peptides than did the corresponding B. abortus OM and LPS. We took advantage of the moderate sensitivity of R B. abortus to cationic peptides to construct live R B. abortus-S-LPS chimeras to test the activities of polymyxin B, lactoferricin B, and EDTA. Homogeneous and abundant peripheral distribution of the heterologous S-LPS was observed on the surface of the chimeras, and this coating had no effect on the viability or morphology of the cells. When the heterologous LPS corresponded to the less sensitive bacterium S B. abortus S19, the chimeras were more resistant to cationic peptides; in contrast, when the S-LPS was from the more sensitive bacterium S. montevideo, the chimeras were more susceptible to the action of peptides and EDTA. A direct correlation between the amount of heterologous S-LPS on the surface of chimeric Brucella cells and peptide sensitivity was observed. Whereas the damage produced by polymyxin B in S. montevideo and B. abortus-S. montevideo S-LPS chimeras was manifested mainly as OM blebbing and inner membrane rolling, lactoferricin B caused inner membrane detachment, vacuolization, and the formation of internal electron-dense granules in these cells. Native S and R B. abortus strains were permeable to the hydrophobic probe N-phenyl-1-naphthylamine (NPN). In contrast, only reduced amounts of NPN partitioned into the OMs of the S. montevideo and B. abortus-S. montevideo S-LPS chimeras. Following peptide exposure, accelerated NPN uptake similar to that observed for S. montevideo was detected for the B. abortus-S. montevideo LPS chimeras. The partition of NPN into native or EDTA-, polymyxin B-, or

  19. Role of urease in megasome formation and Helicobacter pylori survival in macrophages

    PubMed Central

    Schwartz, Justin T.; Allen, Lee-Ann H.

    2007-01-01

    Previous studies have demonstrated that Helicobacter pylori (Hp) delays its entry into macrophages and persists inside megasomes, which are poorly acidified and accumulate early endosome autoantigen 1. Herein, we explored the role of Hp urease in bacterial survival in murine peritoneal macrophages and J774 cells. Plasmid-free mutagenesis was used to replace ureA and ureB with cat in Hp Strains 11637 and 11916. ureAB null Hp lacked detectable urease activity and did not express UreA or UreB as judged by immunoblotting. Deletion of ureAB had no effect on Hp binding to macrophages or the rate or extent of phagocytosis. However, intracellular survival of mutant organisms was impaired significantly. Immunofluorescence microscopy demonstrated that (in contrast to parental organisms) mutant Hp resided in single phagosomes, which were acidic and accumulated the lysosome marker lysosome-associated membrane protein-1 but not early endosome autoantigen 1. A similar phenotype was observed for spontaneous urease mutants derived from Hp Strain 60190. Treatment of macrophages with bafilomycin A1, NH4Cl, or chloroquine prevented acidification of phagosomes containing mutant Hp. However, only ammonium chloride enhanced bacterial viability significantly. Rescue of ureAB null organisms was also achieved by surface adsorption of active urease. Altogether, our data indicate a role for urease and urease-derived ammonia in megasome formation and Hp survival. PMID:16543403

  20. Spatial analysis on China's regional air pollutants and CO2 emissions: emission pattern and regional disparity

    NASA Astrophysics Data System (ADS)

    Dong, Liang; Liang, Hanwei

    2014-08-01

    China has suffered from serious air pollution and CO2 emission. Challenges of emission reduction policy not only come from technology advancement, but also generate from the fact that, China has pronounced disparity between regions, in geographical and socioeconomic. How to deal with regional disparity is important to achieve the reduction target effectively and efficiently. This research conducts a spatial analysis on the emission patterns of three air pollutants named SO2, NOx and PM2.5, and CO2, in China's 30 provinces, applied with spatial auto-correlation and multi regression modeling. We further analyze the regional disparity and inequity issues with the approach of Lorenz curve and Gini coefficient. Results highlight that: there is evident cluster effect for the regional air pollutants and CO2 emissions. While emission amount increases from western regions to eastern regions, the emission per GDP is in inverse trend. The Lorenz curve shows an even larger unequal distribution of GDP/emissions than GDP/capita in 30 regions. Certain middle and western regions suffers from a higher emission with lower GDP, which reveal the critical issue of emission leakage. Future policy making to address such regional disparity is critical so as to promote the emission control policy under the “equity and efficiency” principle.

  1. Examining and contrasting the cognitive activities engaged in undergraduate research experiences and lab courses

    NASA Astrophysics Data System (ADS)

    Holmes, N. G.; Wieman, Carl E.

    2016-12-01

    While the positive outcomes of undergraduate research experiences (UREs) have been extensively categorized, the mechanisms for those outcomes are less understood. Through lightly structured focus group interviews, we have extracted the cognitive tasks that students identify as engaging in during their UREs. We also use their many comparative statements about their coursework, especially lab courses, to evaluate their experimental physics-related cognitive tasks in those environments. We find there are a number of cognitive tasks consistently encountered in physics UREs that are present in most experimental research. These are seldom encountered in lab or lecture courses, with some notable exceptions. Having time to reflect and fix or revise, and having a sense of autonomy, were both repeatedly cited as key enablers of the benefits of UREs. We also identify tasks encountered in actual experimental research that are not encountered in UREs. We use these findings to identify opportunities for better integration of the cognitive tasks in UREs and lab courses, as well as discussing the barriers that exist. This work responds to extensive calls for science education to better develop students' scientific skills and practices, as well as calls to expose more students to scientific research.

  2. Cross-Reactivity of Polyclonal Antibodies against Canavalia ensiformis (Jack Bean) Urease and Helicobacter pylori Urease Subunit A Fragments.

    PubMed

    Kaminski, Zbigniew Jerzy; Relich, Inga; Konieczna, Iwona; Kaca, Wieslaw; Kolesinska, Beata

    2018-01-01

    Overlapping decapeptide fragments of H. pylori urease subunit A (UreA) were synthesized and tested with polyclonal antibodies against Canavalia ensiformis (Jack bean) urease. The linear epitopes of UreA identified using the dot blot method were then examined using epitope mapping. For this purpose, series of overlapping fragments of UreA, frameshifted ± four amino acid residues were synthesized. Most of the UreA epitopes which reacted with the Jack bean urease polyclonal antibodies had been recognized in previous studies by monoclonal antibodies against H. pylori urease. Fragments 11 - 24, 21 - 33, and 31 - 42 were able to interact with the Jack bean urease antibodies, giving stable immunological complexes. However, the lack of recognition by these antibodies of all the components in the peptide map strongly suggests that a non-continuous (nonlinear) epitope is located on the N-terminal domain of UreA. © 2018 Wiley-VHCA AG, Zurich, Switzerland.

  3. MULTILOCUS SEQUENCE TYPING OF BRUCELLA ISOLATES FROM THAILAND.

    PubMed

    Chawjiraphan, Wireeya; Sonthayanon, Piengchan; Chanket, Phanita; Benjathummarak, Surachet; Kerdsin, Anusak; Kalambhaheti, Thareerat

    2016-11-01

    Although brucellosis outbreaks in Thailand are rare, they cause abortions and infertility in animals, resulting in significant economic loss. Because Brucella spp display > 90% DNA homology, multilocus sequence typing (MLST) was employed to categorize local Brucella isolates into sequence types (STs) and to determine their genetic relatedness. Brucella samples were isolated from vaginal secretion of cows and goats, and from blood cultures of infected individuals. Brucella species were determined by multiplex PCR of eight loci, in addition to MLST based on partial DNA sequences of nine house-keeping genes. MLST analysis of 36 isolates revealed 78 distinct novel allele types and 34 novel STs, while two isolates possessed the known ST8. Sequence alignments identified polymorphic sites in each allele, ranging from 2-6%, while overall genetic diversity was 3.6%. MLST analysis of the 36 Brucella isolates classified them into three species, namely, B. melitensis, B. abortus and B. suis, in agreement with multiplex PCR results. Genetic relatedness among ST members of B. melitensis and B. abortus determined by eBURST program revealed ST2 as founder of B. abortus isolates and ST8 the founder of B. melitensis isolates. ST 36, 41 and 50 of Thai Brucella isolates were identified as single locus variants of clonal cluster (CC) 8, while the majority of STs were diverse. The genetic diversity and relatedness identified using MLST revealed hitherto unexpected diversity among Thai Brucella isolates. Genetic classification of isolates could reveal the route of brucellosis transmission among humans and farm animals and also reveal their relationship with other isolates in the region and other parts of the world.

  4. Polymorphonuclear Neutrophils Are Necessary for the Recruitment of CD8+ T Cells in the Liver in a Pregnant Mouse Model of Chlamydophila abortus (Chlamydia psittaci Serotype 1) Infection

    PubMed Central

    de Oca, Roberto Montes; Buendía, Antonio J.; Del Río, Laura; Sánchez, Joaquín; Salinas, Jesús; Navarro, Jose A.

    2000-01-01

    The role of polymorphonuclear neutrophils (PMNs) in the development of the specific immune response against Chlamydophila abortus (Chlamydia psittaci serotype 1) infection was studied in a pregnant mouse model involving treatment with RB6-8C5 monoclonal antibody. PMN depletion significantly affected the immune response in the liver, in which the T-lymphocyte and F4/80+ cell populations decreased, particularly the CD8+ T-cell population. A Th1-like response, characterized by high levels of gamma interferon without detectable levels of interleukin 4 (IL-4) in serum, was observed in both depleted and nondepleted mice, although an increased production of IL-10 was detected in the depleted group. Our results suggest that PMNs play a very important role in the recruitment of other leukocyte populations to the inflammatory foci but have little influence in the polarization of the immune specific response toward a Th1-like response. PMID:10679002

  5. Three enhancer regions regulate gbx2 gene expression in the isthmic region during zebrafish development.

    PubMed

    Islam, Md Ekramul; Kikuta, Hiroshi; Inoue, Fumitaka; Kanai, Maiko; Kawakami, Atsushi; Parvin, Mst Shahnaj; Takeda, Hiroyuki; Yamasu, Kyo

    2006-12-01

    In vertebrate embryos, positioning of the boundary between the midbrain and hindbrain (MHB) and subsequent isthmus formation are dependent upon the interaction between the Otx2 and Gbx genes. In zebrafish, sequential expression of gbx1 and gbx2 in the anterior hindbrain contributes to this process, whereas in mouse embryos, a single Gbx gene (Gbx2) is responsible for MHB development. In the present study, to investigate the regulatory mechanism of gbx2 in the MHB/isthmic region of zebrafish embryos, we cloned the gene and showed that its organization is conserved among different vertebrates. Promoter analyses revealed three enhancers that direct reporter gene expression after the end of epiboly in the anterior-most hindbrain, which is a feature of the zebrafish gbx2 gene. One of the enhancers is located upstream of gbx2 (AMH1), while the other two enhancers are located downstream of gbx2 (AMH2 and AMH3). Detailed analysis of the AMH1 enhancer showed that it directs expression in the rhombomere 1 (r1) region and the dorsal thalamus, as has been shown for gbx2, whereas no expression was induced by the AMH1 enhancer in other embryonic regions in which gbx2 is expressed. The AMH1 enhancer is composed of multiple regulatory subregions that share the same spatial specificity. The most active of the regulatory subregions is a 291-bp region that contains at least two Pax2-binding sites, both of which are necessary for the function of the main component (PB1-A region) of the AMH1 enhancer. In accordance with these results, enhancer activity in the PB1-A region, as well as gbx2 expression in r1, was missing in no isthmus mutant embryos that lacked functional pax2a. In addition, we identified an upstream conserved sequence of 227bp that suppresses the enhancer activity of AMH1. Taken together, these findings suggest that gbx2 expression during the somitogenesis stage in zebrafish is regulated by a complex mechanism involving Pax2 as well as activators and suppressors in the

  6. BvrR/BvrS-Controlled Outer Membrane Proteins Omp3a and Omp3b Are Not Essential for Brucella abortus Virulence▿

    PubMed Central

    Manterola, Lorea; Guzmán-Verri, Caterina; Chaves-Olarte, Esteban; Barquero-Calvo, Elías; de Miguel, María-Jesús; Moriyón, Ignacio; Grilló, María-Jesús; López-Goñi, Ignacio; Moreno, Edgardo

    2007-01-01

    The Brucella abortus two-component regulatory system BvrR/BvrS controls the expression of outer membrane proteins (Omp) Omp3a (Omp25) and Omp3b (Omp22). Disruption of bvrS or bvrR generates avirulent mutants with altered cell permeability, higher sensitivity to microbicidal peptides, and complement. Consequently, the role of Omp3a and Omp3b in virulence was examined. Similar to bvrS or bvrR mutants, omp3a and omp3b mutants displayed increased attachment to cells, indicating surface alterations. However, they showed unaltered permeability; normal expression of Omp10, Omp16, Omp19, Omp2b, and Omp1; native hapten polysaccharide; and lipopolysaccharide and were resistant to complement and polymyxin B at ranges similar to those of the wild-type (WT) counterpart. Likewise, omp3a and omp3b mutants were able to replicate in murine macrophages and in HeLa cells, were resistant to the killing action of human neutrophils, and persisted in mice, like the WT strain. Murine macrophages infected with the omp3a mutant generated slightly higher levels of tumor necrosis factor alpha than the WT, whereas the bvrS mutant induced lower levels of this cytokine. Since the absence of Omp3a or Omp3b does not result in attenuation, it can be concluded that BvrR/BvrS influences additional Brucella properties involved in virulence. Our results are discussed in the light of previous works suggesting that disruption of omp3a generates attenuated Brucella strains, and we speculate on the role of group 3 Omps. PMID:17664262

  7. Yeast prions assembly and propagation

    PubMed Central

    2011-01-01

    Yeast prions are self-perpetuating protein aggregates that are at the origin of heritable and transmissible non-Mendelian phenotypic traits. Among these, [PSI+], [URE3] and [PIN+] are the most well documented prions and arise from the assembly of Sup35p, Ure2p and Rnq1p, respectively, into insoluble fibrillar assemblies. Fibril assembly depends on the presence of N- or C-terminal prion domains (PrDs) which are not homologous in sequence but share unusual amino-acid compositions, such as enrichment in polar residues (glutamines and asparagines) or the presence of oligopeptide repeats. Purified PrDs form amyloid fibrils that can convert prion-free cells to the prion state upon transformation. Nonetheless, isolated PrDs and full-length prion proteins have different aggregation, structural and infectious properties. In addition, mutations in the “non-prion” domains (non-PrDs) of Sup35p, Ure2p and Rnq1p were shown to affect their prion properties in vitro and in vivo. Despite these evidences, the implication of the functional non-PrDs in fibril assembly and prion propagation has been mostly overlooked. In this review, we discuss the contribution of non-PrDs to prion assemblies, and the structure-function relationship in prion infectivity in the light of recent findings on Sup35p and Ure2p assembly into infectious fibrils from our laboratory and others. PMID:22052349

  8. Effect of heavy metals on soil enzyme activity at different field conditions in Middle Spis mining area (Slovakia).

    PubMed

    Angelovičová, Lenka; Lodenius, Martin; Tulisalo, Esa; Fazekašová, Danica

    2014-12-01

    Heavy metals concentrations were measured in the former mining area located in Hornad river valley (Slovakia). Soil samples were taken in 2012 from 20 sites at two field types (grasslands, heaps of waste material) and two different areas. Total content of heavy metals (Cu, Pb, Zn, Hg), urease (URE), acid phosphatase (ACP), alkaline phosphatase (ALP), soil reaction (pH) were changing depending on the field/area type. The tailing pond and processing plants have been found as the biggest sources of pollution. URE, ACP and ALP activities significantly decreased while the heavy metal contents increased. Significant differences were found among area types in the heavy metal contents and activity of URE. No statistical differences in the content of heavy metals but significant statistical differences for soil pH were found for field types (grassland and heaps). Significant negative correlation was found for URE-Pb, URE-Zn and also between soil reaction and ACP and ALP.

  9. Identification Of Protein Vaccine Candidates Using Comprehensive Proteomic Analysis Strategies

    DTIC Science & Technology

    2007-12-01

    urease (URE) gene codes for a urea amidohydrolase protein that catalyzes urea hydrolysis. The protein was first isolated from C. immitis and...the Cu, Zn, Superoxide Dismutase (SOD), the Spherule Outer Wall glycoprotein (SOWgp), the T-Cell Reactive Protein (TCRP), and Urease (URE). It is...et al. 1997. Isolation and characterization of the urease gene (URE) from the pathogenic fungus Coccidioides immitis. Gene 198: 387-391. 54. Li, K

  10. Anti-neuropathic effects of Rosmarinus officinalis L. terpenoid fraction: relevance of nicotinic receptors

    PubMed Central

    Mannelli, Lorenzo Di Cesare; Micheli, Laura; Maresca, Mario; Cravotto, Giancarlo; Bellumori, Maria; Innocenti, Marzia; Mulinacci, Nadia; Ghelardini, Carla

    2016-01-01

    Traditional uses and current results highlight the neuroprotective properties of Rosmarinus officinalis L. The compelling need for novel strategies able to relieve neuropathic pain encouraged us to analyze different rosemary leaf extracts in rats following chronic constriction injury (CCI) of sciatic nerve. Ethanol, acetone, and the innovative ultrasound-hexane extractive methods were used to obtain: EE, AE, and for hexane extracts UREprel and URE. Extracts were characterized in terms of typical constituents and repeatedly administered to CCI-rats (13-days treatment, from the day of surgery). URE showed the best efficacy and potency in reducing hypersensitivity to noxious- and non-noxious stimuli and spontaneous pain. URE contained the higher quantity of the terpenoid carnosic acid (CA) and its efficacy was compared to pure CA. Histological analysis of the sciatic nerve revealed that URE prevented axon and myelin derangement, edema and inflammatory infiltrate. In the dorsal horn of the spinal cord, URE did not reduce astrocyte activation. Both the pain reliever and the neuroconservative effects of URE were significantly prevented by the nicotinic receptor (nAChR) antagonist mecamylamine. In conclusion, the hexane-ultrasound rosemary extract is able to reduce neuropathic hypersensitivity and protect nervous tissues. Effectiveness is mainly related to the terpenoid fraction by mechanisms involving nAChRs. PMID:27713514

  11. Novel characteristics of the biological properties of the yeast Saccharomyces cerevisiae eukaryotic initiation factor 2A.

    PubMed

    Komar, Anton A; Gross, Stephane R; Barth-Baus, Diane; Strachan, Ryan; Hensold, Jack O; Goss Kinzy, Terri; Merrick, William C

    2005-04-22

    Eukaryotic initiation factor 2A (eIF2A) has been shown to direct binding of the initiator methionyl-tRNA (Met-tRNA(i)) to 40 S ribosomal subunits in a codon-dependent manner, in contrast to eIF2, which requires GTP but not the AUG codon to bind initiator tRNA to 40 S subunits. We show here that yeast eIF2A genetically interacts with initiation factor eIF4E, suggesting that both proteins function in the same pathway. The double eIF2A/eIF4E-ts mutant strain displays a severe slow growth phenotype, which correlated with the accumulation of 85% of the double mutant cells arrested at the G(2)/M border. These cells also exhibited a disorganized actin cytoskeleton and elevated actin levels, suggesting that eIF2A might be involved in controlling the expression of genes involved in morphogenic processes. Further insights into eIF2A function were gained from the studies of eIF2A distribution in ribosomal fractions obtained from either an eIF5BDelta (fun12Delta) strain or a eIF3b-ts (prt1-1) strain. It was found that the binding of eIF2A to 40 and 80 S ribosomes was not impaired in either strain. We also found that eIF2A functions as a suppressor of Ure2p internal ribosome entry site-mediated translation in yeast cells. The regulation of expression from the URE2 internal ribosome entry site appears to be through the levels of eIF2A protein, which has been found to be inherently unstable with a half-life of approximately 17 min. It was hypothesized that this instability allows for translational control through the level of eIF2A protein in yeast cells.

  12. 13 CFR 300.2 - EDA Headquarters and regional offices.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 13 Business Credit and Assistance 1 2010-01-01 2010-01-01 false EDA Headquarters and regional offices. 300.2 Section 300.2 Business Credit and Assistance ECONOMIC DEVELOPMENT ADMINISTRATION, DEPARTMENT OF COMMERCE GENERAL INFORMATION § 300.2 EDA Headquarters and regional offices. (a) EDA's...

  13. The Use of Case History Studies to Differentiate Potentially Infected from Potentially Noninfected Herds with Reactors to Brucella abortus Antigens

    PubMed Central

    Martin, S. W.; Gerrow, A. F.

    1978-01-01

    Data on farm characteristics and the results of the first herd test for brucellosis were collected for 74 reactor and 74 negative herds in Wellington County, Ontario. Each reactor herd was classified as either probably infected or probably not infected using the occurrence of abortions prior to the first herd test or during the testing period, the total number of cattle removed and/or the spread of reactors within the herd as criteria of infection. Statistical techniques were used to select variables which were good “discriminators” between probably infected and noninfected herds. In general, reactor herds were primarily dairy herds and were somewhat larger than negative herds. The presence of only single suspicious reactors on the first test appeared to be a good predictor of lack of infection with Brucella abortus. Among the 37 farms in this category the single reactor was removed from only eight farms and no evidence o fthe spread of infection was observed. The presence of one or more positive reactors on the first herd test appeared to be a good predictor of the presence of infection. Of the 24 farms in this category, evidence of the spread of infection was present in ten farms and seven of these ten farms were eventually depopulated. The brucella milk ring test appeared to be the most effective means of identifying infected herds under the conditions present in Wellington County. PMID:417777

  14. The Lipopolysaccharide of Brucella abortus BvrS/BvrR Mutants Contains Lipid A Modifications and Has Higher Affinity for Bactericidal Cationic Peptides

    PubMed Central

    Manterola, Lorea; Moriyón, Ignacio; Moreno, Edgardo; Sola-Landa, Alberto; Weiss, David S.; Koch, Michel H. J.; Howe, Jörg; Brandenburg, Klaus; López-Goñi, Ignacio

    2005-01-01

    The two-component BvrS/BvrR system is essential for Brucella abortus virulence. It was shown previously that its dysfunction abrogates expression of some major outer membrane proteins and increases bactericidal peptide sensitivity. Here, we report that BvrS/BvrR mutants have increased surface hydrophobicity and susceptibility to killing by nonimmune serum. The bvrS and bvrR mutant lipopolysaccharides (LPSs) bound more polymyxin B, chimeras constructed with bvrS mutant cells and parental LPS showed augmented polymyxin B resistance, and, conversely, parental cells and bvrS mutant LPS chimeras were more sensitive and displayed polymyxin B-characteristic outer membrane lesions, implicating LPS as being responsible for the phenotype of the BvrS/BvrR mutants. No qualitative or quantitative changes were detected in other envelope and outer membrane components examined: periplasmic β(1-2) glucans, native hapten polysaccharide, and phospholipids. The LPS of the mutants was similar to parental LPS in O-polysaccharide polymerization and fine structure but showed both increased underacylated lipid A species and higher acyl-chain fluidity that correlated with polymyxin B binding. These lipid A changes did not alter LPS cytokine induction, showing that in contrast to other gram-negative pathogens, recognition by innate immune receptors is not decreased by these changes in LPS structure. Transcription of Brucella genes required for incorporating long acyl chains into lipid A (acpXL and lpxXL) or implicated in lipid A acylation control (bacA) was not affected. We propose that in Brucella the outer membrane homeostasis depends on the functioning of BvrS/BvrR. Accordingly, disruption of BvrS/BvrR damages the outer membrane, thus contributing to the severe attenuation manifested by bvrS and bvrR mutants. PMID:16077108

  15. Characterization of Genomic Island 3 and Genetic Variability of Chilean Field Strains of Brucella abortus▿

    PubMed Central

    Céspedes, Sandra; Salgado, Paulina; Valenzuela, Patricio; Vidal, Roberto; Oñate, Angel A.

    2011-01-01

    One of the capabilities developed by bacteria is the ability to gain large fragments of DNA from other bacteria or to lose portions of their own genomes. Among these exchangeable fragments are the genomic islands (GIs). Nine GIs have been identified in Brucella, and genomic island 3 (GI-3) is shared by two pathogenic species, B. melitensis and B. abortus. GI-3 encodes mostly unknown proteins. One of the aims of this study was to perform pulsed-field gel electrophoresis (PFGE) on field isolates of B. abortus from Chile to determine whether these isolates are clonally related. Furthermore, we focused on the characterization of GI-3, studying its organization and the genetic conservation of the GI-3 sequence using techniques such as tiling-path PCR (TP-PCR) and restriction fragment length polymorphism-PCR (RFLP-PCR). Our results, after PFGE was performed on 69 field isolates of B. abortus from Chile, showed that the strains were genetically homogeneous. To increase the power of genetic discrimination among these strains, we used multiple locus variable-number tandem-repeat (VNTR) analysis with 16 loci (MLVA-16). The results obtained by MLVA-16 showed that the strains of B. abortus were genetically heterogeneous and that most of them clustered according to their geographic origin. Of the genetic loci studied, panel 2B was the one describing the highest diversity in the analysis, as well as locus Bruce19 in panel 2A. In relation to the study of GI-3, our experimental analysis by TP-PCR identified and confirmed that GI-3 is present in all wild strains of B. abortus, demonstrating the high stability of gene cluster GI-3 in Chilean field strains. PMID:21543580

  16. 1,2-Benzisoselenazol-3(2H)-one Derivatives As a New Class of Bacterial Urease Inhibitors.

    PubMed

    Macegoniuk, Katarzyna; Grela, Ewa; Palus, Jerzy; Rudzińska-Szostak, Ewa; Grabowiecka, Agnieszka; Biernat, Monika; Berlicki, Łukasz

    2016-09-08

    Urease inhibitors are considered promising compounds for the treatment of ureolytic bacterial infections, particularly infections resulting from Helicobacter pylori in the gastric tract. Herein, we present the synthesis and the inhibitory activity of novel and highly effective organoselenium compounds as inhibitors of Sporosarcina pasteurii and Helicobacter pylori ureases. These studied compounds represent a class of competitive reversible urease inhibitors. The most active compound, 2-phenyl-1,2-benzisoselenazol-3(2H)-one (ebselen), displayed Ki values equal to 2.11 and 226 nM against S. pasteurii and H. pylori enzymes, respectively, indicating ebselen as one of the most potent low-molecular-weight inhibitors of bacterial ureases reported to date. Most of these molecules penetrated through the cell membrane of the Gram-negative bacteria Escherichia coli (pGEM::ureOP) in vitro. Furthermore, whole-cell studies on the H. pylori J99 reference strain confirmed the high efficiency of the examined organoselenium compounds as urease inhibitors against pathogenic bacteria.

  17. Source regions and transport pathways of PM2.5 at a regional background site in East China

    NASA Astrophysics Data System (ADS)

    Zhang, Yanru; Zhang, Hongliang; Deng, Junjun; Du, Wenjiao; Hong, Youwei; Xu, Lingling; Qiu, Yuqing; Hong, Zhenyu; Wu, Xin; Ma, Qianli; Yao, Jie; Chen, Jinsheng

    2017-10-01

    PM2.5 samples were collected daily at the Lin'an regional background station (LA) in Zhejiang, China during 2014-2015 and the major chemical components including organic carbon (OC), elemental carbon (EC) and water-soluble inorganic ions (WSII) were determined. Backward trajectory clustering and potential source contribution function (PSCF) were adopted for identifying the transport pathways and potential source regions of PM2.5 at LA. The annual mean concentration was 68.9 ± 28.3 μg m-3, indicating severe pollution in East China. Obvious seasonal variations were found, with highest level in winter and lowest in summer. Carbonaceous aerosols and WSII were the predominant compositions, accounting for 30.7% and 53.5% of PM2.5, respectively. Secondary inorganic ions (SO42-, NO3-, and NH4+) made a total contribution of 45.2% to PM2.5. Heterogeneous formation played a dominant role in SO42- formation and NH4+ formation promoted NO3- formation. Stationary sources played a more important role than mobile sources based on NO3-/SO42- ratio of 0.53. Aerosol environment at LA was ammonium-poor and SO42- was only neutralized sufficiently by NH4+ with the predominant production of (NH4)2SO4 in winter. Four major transport pathways of air masses at LA were found based on trajectory clustering. Air masses from the northwest Gobi areas passing over the heavily polluted regions in North and Central China had the highest levels of PM2.5, followed by the air masses from Central China. PSCF results suggested that surrounding areas in the Yangtze River Delta region were major regional origins of PM2.5 and its major components. Northern region was an important origin for carbonaceous components, and southwestern region was significant for secondary inorganic ions. This study helps understand PM2.5 characteristics, identify potential source regions and effectively control PM2.5 in East China.

  18. Helicobacter pylori Urease Activity is Influenced by Ferric Uptake Regulator

    PubMed Central

    Lee, Jong Seung; Lee, Ji Hyuk; Lee, Hye Jin; Lee, Jee Hyun; Choi, Young Ok

    2010-01-01

    Purpose The role of the Ferric Uptake Regulator (FUR) in the acid resistance of Helicobacter pylori (H. pylori) has been thought to be independent of urease. However, we demonstrated in this study that Fur influences urease activity. Materials and Methods A fur knockout mutant of H. pylori was constructed by replacing the Fur gene with a kanamycin resistant marker gene. The wild-type H. pylori and fur mutant were compared for survival. The integrity of the inner membrane of the bacteria was evaluated by confocal microscopy using membrane-permeant and -impermeant fluorescent DNA probes. Urease activity of intact H. pylori was measured between pH 3 and 8. Real time PCR of both strains was performed for urease genes including ureI, ureE, ureF, ureG, and ureH. Results The fur deletion affected the survival of H. pylori at pH 4. The urease activity curve of the intact fur mutant showed the same shape as the wild-type but was 3-fold lower than the wild-type at a pH of less than 5. Real time PCR revealed that the expression of all genes was consistently down-regulated in the fur mutant. Conclusion The results of this study showed that fur appears to be involved in acid resistant H. pylori urease activity. PMID:20046512

  19. Prevalence of uncorrected refractive errors among children aged 3-10 years in western Saudi Arabia

    PubMed Central

    Alrahili, Nojood Hameed R.; Jadidy, Esraa S.; Alahmadi, Bayan Sulieman H.; Abdula’al, Mohammed F.; Jadidy, Alaa S.; Alhusaini, Abdulaziz A.; Mojaddidi, Moaz A.; Al-Barry, Maan A.

    2017-01-01

    Objectives: To determine the prevalence of uncorrected refractive errors (URE) among children 3-10 years and to affirm the necessity of a national school-based visual screening program for school-aged children. Methods: This retrospective cross-sectional study was conducted in Medina, Saudi Arabia in 2015. Children were selected through a multistage stratified random sampling from 8 kindergarten and 8 primary schools. Those included were screened to diagnose UREs using a visual acuity chart and an auto refractometer according to American guidelines. The prevalence and types of UREs were estimated. Results: Of the 2121 children enumerated, 1893 were examined, yielding a response rate of 89.3%. The prevalence of UREs was 34.9% (95% CI = 32.8%-37.1%), with significant differences in different age groups. The prevalence of astigmatism (25.3%) was higher compared to that of anisometropia (7.4%), hypermetropia (1.5%), and myopia (0.7%). Risk of uncorrected refractive error was positively associated with age, and this was noted in astigmatism, myopia, and anisometropia. In addition, the risk of hypermetropia was associated with boys and that of myopia was associated with girls. Conclusions: The prevalence of UREs, particularly astigmatism, was high among children aged 3-10 years in Medina, with significant age differences. Vision screening programs targeting kindergarten and primary schoolchildren are crucial to lessen the risk of preventable visual impairment due to UREs. PMID:28762432

  20. Prevalence of uncorrected refractive errors among children aged 3-10 years in western Saudi Arabia.

    PubMed

    Alrahili, Nojood Hameed R; Jadidy, Esraa S; Alahmadi, Bayan Sulieman H; Abdula'al, Mohammed F; Jadidy, Alaa S; Alhusaini, Abdulaziz A; Mojaddidi, Moaz A; Al-Barry, Maan A

    2017-08-01

    To determine the prevalence of uncorrected refractive errors (URE) among children 3-10 years and to affirm the necessity of a national school-based visual screening program for school-aged children. Methods: This retrospective cross-sectional study was conducted in Medina, Saudi Arabia in 2015. Children were selected through a multistage stratified random sampling from 8 kindergarten and 8 primary schools. Those included were screened to diagnose UREs using a visual acuity chart and an auto refractometer according to American guidelines. The prevalence and types of UREs were estimated. Results: Of the 2121 children enumerated, 1893 were examined, yielding a response rate of 89.3%. The prevalence of UREs was 34.9% (95% CI = 32.8%-37.1%), with significant differences in different age groups. The prevalence of astigmatism (25.3%) was higher compared to that of anisometropia (7.4%), hypermetropia (1.5%), and myopia (0.7%). Risk of uncorrected refractive error was positively associated with age, and this was noted in astigmatism, myopia, and anisometropia. In addition, the risk of hypermetropia was associated with boys and that of myopia was associated with girls. Conclusions: The prevalence of UREs, particularly astigmatism, was high among children aged 3-10 years in Medina, with significant age differences. Vision screening programs targeting kindergarten and primary schoolchildren are crucial to lessen the risk of preventable visual impairment due to UREs.

  1. A Dimensionally Aligned Signal Projection for Classification of Unintended Radiated Emissions

    DOE PAGES

    Vann, Jason Michael; Karnowski, Thomas P.; Kerekes, Ryan; ...

    2017-04-24

    Characterization of unintended radiated emissions (URE) from electronic devices plays an important role in many research areas from electromagnetic interference to nonintrusive load monitoring to information system security. URE can provide insights for applications ranging from load disaggregation and energy efficiency to condition-based maintenance of equipment-based upon detected fault conditions. URE characterization often requires subject matter expertise to tailor transforms and feature extractors for the specific electrical devices of interest. We present a novel approach, named dimensionally aligned signal projection (DASP), for projecting aligned signal characteristics that are inherent to the physical implementation of many commercial electronic devices. These projectionsmore » minimize the need for an intimate understanding of the underlying physical circuitry and significantly reduce the number of features required for signal classification. We present three possible DASP algorithms that leverage frequency harmonics, modulation alignments, and frequency peak spacings, along with a two-dimensional image manipulation method for statistical feature extraction. To demonstrate the ability of DASP to generate relevant features from URE, we measured the conducted URE from 14 residential electronic devices using a 2 MS/s collection system. Furthermore, a linear discriminant analysis classifier was trained using DASP generated features and was blind tested resulting in a greater than 90% classification accuracy for each of the DASP algorithms and an accuracy of 99.1% when DASP features are used in combination. Furthermore, we show that a rank reduced feature set of the combined DASP algorithms provides a 98.9% classification accuracy with only three features and outperforms a set of spectral features in terms of general classification as well as applicability across a broad number of devices.« less

  2. A Dimensionally Aligned Signal Projection for Classification of Unintended Radiated Emissions

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Vann, Jason Michael; Karnowski, Thomas P.; Kerekes, Ryan

    Characterization of unintended radiated emissions (URE) from electronic devices plays an important role in many research areas from electromagnetic interference to nonintrusive load monitoring to information system security. URE can provide insights for applications ranging from load disaggregation and energy efficiency to condition-based maintenance of equipment-based upon detected fault conditions. URE characterization often requires subject matter expertise to tailor transforms and feature extractors for the specific electrical devices of interest. We present a novel approach, named dimensionally aligned signal projection (DASP), for projecting aligned signal characteristics that are inherent to the physical implementation of many commercial electronic devices. These projectionsmore » minimize the need for an intimate understanding of the underlying physical circuitry and significantly reduce the number of features required for signal classification. We present three possible DASP algorithms that leverage frequency harmonics, modulation alignments, and frequency peak spacings, along with a two-dimensional image manipulation method for statistical feature extraction. To demonstrate the ability of DASP to generate relevant features from URE, we measured the conducted URE from 14 residential electronic devices using a 2 MS/s collection system. Furthermore, a linear discriminant analysis classifier was trained using DASP generated features and was blind tested resulting in a greater than 90% classification accuracy for each of the DASP algorithms and an accuracy of 99.1% when DASP features are used in combination. Furthermore, we show that a rank reduced feature set of the combined DASP algorithms provides a 98.9% classification accuracy with only three features and outperforms a set of spectral features in terms of general classification as well as applicability across a broad number of devices.« less

  3. Bacterial, fungal, parasitological and pathological analyses of abortions in small ruminants from 2012-2016.

    PubMed

    Schnydrig, P; Vidal, S; Brodard, I; Frey, C; Posthaus, H; Perreten, V; Rodriguez-Campos, S

    2017-12-01

    Abortion in small ruminants presents a clinical and economic problem with legal implications regarding animal health and zoonotic risk by some of the abortive pathogens. Several bacteria, fungi and parasites can cause abortion, but cost-orientated routine diagnostics only cover the most relevant epizootic agents. To cover a broad-range of common as well as underdiagnosed abortifacients, we studied 41 ovine and 36 caprine abortions by Stamp's modification of the Ziehl-Neelsen stain, culture for classical and opportunistic abortive agents, real-time PCR for C. burnetii, C. abortus, pathogenic Leptospira spp., Toxoplasma gondii and Neospora caninum. When the dam's serum was available detection of antibodies against B. melitensis, C. burnetii, C. abortus and Leptospira spp. was performed. In 37 cases sufficient placental tissue was available for pathological and histopathological examination. From the 77 cases 11 (14.3%) were positive by staining whereas real-time PCR detected C. burnetii and C. abortus in 49.3% and 32.5% of the cases. Antibodies against C. abortus and Leptospira spp. (33.3 and 26.7%) were detected. In 23.4% a bacterial culturable pathogen was isolated. Fungal abortion was confirmed in 1.3% of cases. A single abortive agent was identified in 44.2% of the cases and in 31.2% multiple possible abortifacients were present. Our study shows that the highest clarification rate can only be achieved by a combination of methods and evidences the role that multi-infections play as cause of abortion.

  4. [Isolation of Escherichia coli O128:HNM harboring stx2f gene from diarrhea patients].

    PubMed

    Isobe, Junko; Kimata, Keiko; Shimojima, Masahiro; Hosorogi, Shiho; Tanaka, Daisuke; Gyobu, Yotaku

    2004-12-01

    Shiga-like-toxin-producing Esherichia coli O128:HNM were isolated from feces of a one-year-old boy with diarrhea and abdominal pain on July, 2002, and a 11-month-old girl with diarrhea and fever on June, 1997. None of other enteropathogenic bacteria including Salmonella were isolated. E. coli O128:HNM isolates from both patients carry stx2f and eaeA gene, but not stx1, stx2, aggR, bfpA, esth, estp, invE, astA, ureC and hlyA gene. As far as we know, this may be the first report indicating that E. coli O128:HNM carrying stx2f gene were isolated from patients in Japan.

  5. 18 CFR 2.21 - Regional Transmission Groups.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 18 Conservation of Power and Water Resources 1 2014-04-01 2014-04-01 false Regional Transmission Groups. 2.21 Section 2.21 Conservation of Power and Water Resources FEDERAL ENERGY REGULATORY COMMISSION, DEPARTMENT OF ENERGY GENERAL RULES GENERAL POLICY AND INTERPRETATIONS Statements of General Policy and...

  6. 18 CFR 2.21 - Regional Transmission Groups.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 18 Conservation of Power and Water Resources 1 2012-04-01 2012-04-01 false Regional Transmission Groups. 2.21 Section 2.21 Conservation of Power and Water Resources FEDERAL ENERGY REGULATORY COMMISSION, DEPARTMENT OF ENERGY GENERAL RULES GENERAL POLICY AND INTERPRETATIONS Statements of General Policy and...

  7. 18 CFR 2.21 - Regional Transmission Groups.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 18 Conservation of Power and Water Resources 1 2013-04-01 2013-04-01 false Regional Transmission Groups. 2.21 Section 2.21 Conservation of Power and Water Resources FEDERAL ENERGY REGULATORY COMMISSION, DEPARTMENT OF ENERGY GENERAL RULES GENERAL POLICY AND INTERPRETATIONS Statements of General Policy and...

  8. 18 CFR 2.21 - Regional Transmission Groups.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 18 Conservation of Power and Water Resources 1 2010-04-01 2010-04-01 false Regional Transmission Groups. 2.21 Section 2.21 Conservation of Power and Water Resources FEDERAL ENERGY REGULATORY COMMISSION, DEPARTMENT OF ENERGY GENERAL RULES GENERAL POLICY AND INTERPRETATIONS Statements of General Policy and...

  9. 50 CFR 2.2 - Regional offices.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... the States of Arizona, New Mexico, Oklahoma, and Texas), 500 Gold Avenue SW., Room 9018 (P.O. Box 1306... States of Colorado, Kansas, Montana, Nebraska, North Dakota, South Dakota, Utah and Wyoming), 134 Union Boulevard (P.O. Box 25486), Lakewood, Colorado 80228. (g) Alaska Regional Office (Region 7—comprising the...

  10. Prevalence of visual impairment due to uncorrected refractive error: Results from Delhi-Rapid Assessment of Visual Impairment Study.

    PubMed

    Senjam, Suraj Singh; Vashist, Praveen; Gupta, Noopur; Malhotra, Sumit; Misra, Vasundhara; Bhardwaj, Amit; Gupta, Vivek

    2016-05-01

    To estimate the prevalence of visual impairment (VI) due to uncorrected refractive error (URE) and to assess the barriers to utilization of services in the adult urban population of Delhi. A population-based rapid assessment of VI was conducted among people aged 40 years and above in 24 randomly selected clusters of East Delhi district. Presenting visual acuity (PVA) was assessed in each eye using Snellen's "E" chart. Pinhole examination was done if PVA was <20/60 in either eye and ocular examination to ascertain the cause of VI. Barriers to utilization of services for refractive error were recorded with questionnaires. Of 2421 individuals enumerated, 2331 (96%) individuals were examined. Females were 50.7% among them. The mean age of all examined subjects was 51.32 ± 10.5 years (standard deviation). VI in either eye due to URE was present in 275 individuals (11.8%, 95% confidence interval [CI]: 10.5-13.1). URE was identified as the most common cause (53.4%) of VI. The overall prevalence of VI due to URE in the study population was 6.1% (95% CI: 5.1-7.0). The elder population as well as females were more likely to have VI due to URE (odds ratio [OR] = 12.3; P < 0.001 and OR = 1.5; P < 0.02). Lack of felt need was the most common reported barrier (31.5%). The prevalence of VI due to URE among the urban adult population of Delhi is still high despite the availability of abundant eye care facilities. The majority of reported barriers are related to human behavior and attitude toward the refractive error. Understanding these aspects will help in planning appropriate strategies to eliminate VI due to URE.

  11. Heterogeneity of D2 dopamine receptors in different brain regions.

    PubMed Central

    Leonard, M N; Macey, C A; Strange, P G

    1987-01-01

    The binding of [3H]spiperone has been examined in membranes derived from different regions of bovine brain. In caudate nucleus, nucleus accumbens, olfactory tubercle and putamen binding is to D2 dopamine and 5HT2 serotonin receptors, whereas in cingulate cortex only serotonin 5HT2 receptor binding can be detected. D2 dopamine receptors were examined in detail in caudate nucleus, olfactory tubercle and putamen using [3H]spiperone binding in the presence of 0.3 microM-mianserin (to block 5HT2 serotonin receptors). No evidence for heterogeneity among D2 dopamine receptors either between brain regions or within a brain region was found from the displacements of [3H]spiperone binding by a range of antagonists, including dibenzazepines and substituted benzamides. Regulation of agonist binding by guanine nucleotides did, however, differ between regions. In caudate nucleus a population of agonist binding sites appeared resistant to guanine nucleotide regulation, whereas this was not the case in olfactory tubercle and putamen. PMID:2963621

  12. Methanolic extracts of Uncaria rhynchophylla induce cytotoxicity and apoptosis in HT-29 human colon carcinoma cells.

    PubMed

    Jo, Kyung-Jin; Cha, Mi-Ran; Lee, Mi-Ra; Yoon, Mi-Young; Park, Hae-Ryong

    2008-06-01

    In this paper, we report the anticancer activities of Uncaria rhynchophylla extracts, a Rubiaceae plant native to China. Traditionally, Uncaria rhynchophylla has been used in the prevention and treatment of neurotoxicity. However, the cytotoxic activity of Uncaria rhynchophylla against human colon carcinoma cells has not, until now, been elucidated. We found that the methanolic extract of Uncaria rhynchophylla (URE) have cytotoxic effects on HT-29 cells. The URE showed highly cytotoxic effects via the MTT reduction assay, LDH release assay, and colony formation assay. As expected, URE inhibited the growth of HT-29 cells in a dose-dependent manner. In particular, the methanolic URE of the 500 microg/ml showed 15.8% inhibition against growth of HT-29 cells. It induced characteristic apoptotic effects in HT-29 cells, including chromatin condensation and sharking occurring 24 h when the cells were treated at a concentration of the 500 microg/ml. The activation of caspase-3 and the specific proteolytic cleavage of poly (ADP-ribose) polymerase were detected over the course of apoptosis induction. These results indicate that URE contains bioactive materials with strong activity, and is a potential chemotherapeutic agent candidate against HT-29 human colon carcinoma cells.

  13. Deuteration around the ultracompact HII region Monoceros R2

    NASA Astrophysics Data System (ADS)

    Treviño-Morales, S. P.; Pilleri, P.; Fuente, A.; Kramer, C.; Roueff, E.; González-García, M.; Cernicharo, J.; Gerin, M.; Goicoechea, J. R.; Pety, J.; Berné, O.; Ossenkopf, V.; Ginard, D.; García-Burillo, S.; Rizzo, J. R.; Viti, S.

    2014-09-01

    Context. The massive star-forming region Monoceros R2 (Mon R2) hosts the closest ultra-compact Hii region, where the photon-dominated region (PDR) between the ionized and molecular gas can be spatially resolved with current single-dish telescopes. Aims: We aim at studying the chemistry of deuterated molecules toward Mon R2 to determine the deuterium fractions around a high-UV irradiated PDR and investigate the chemistry of these species. Methods: We used the IRAM-30 m telescope to carry out an unbiased spectral survey toward two important positions (namely IF and MP2) in Mon R2 at 1, 2, and 3 mm. This spectral survey is the observational basis of our study of the deuteration in this massive star-forming region. Our high spectral resolution observations (~0.25-0.65 km s-1) allowed us to resolve the line profiles of the different species detected. Results: We found a rich chemistry of deuterated species at both positions of Mon R2, with detections of C2D, DCN, DNC, DCO+, D2CO, HDCO, NH2D, and N2D+ and their corresponding hydrogenated species and rarer isotopologs. The high spectral resolution of our observations allowed us to resolve three velocity components: the component at 10 km s-1 is detected at both positions and seems associated with the layer most exposed to the UV radiation from IRS 1; the component at 12 km s-1 is found toward the IF position and seems related to the foreground molecular gas; finally, a component at 8.5 km s-1 is only detected toward the MP2 position, most likely related to a low-UV irradiated PDR. We derived the column density of the deuterated species (together with their hydrogenated counterparts), and determined the deuterium fractions as Dfrac = [XD]/[XH]. The values of Dfrac are around 0.01 for all the observed species, except for HCO+ and N2H+, which have values 10 times lower. The values found in Mon R2 are similar to those measured in the Orion Bar, and are well explained with a pseudo-time-dependent gas-phase model in which

  14. [Pregnancy-specific beta-glycoprotein in the serum of women with a complicated early pregnancy].

    PubMed

    Radikov, N

    1989-01-01

    The author determined pregnancy specific beta 1-glycoprotein in 109 women with threatened early pregnancy as 32 of the women suffered from abortus imminens with several unsuccessful pregnancies in the past as well as 67 women with abortus incipiens with bleeding ex utero. The author established that 87% of women with abortus imminens and preserved pregnancies had values of beta 1-glycoprotein close to those of normal pregnancy for the respective gestational week. 93% of women with abortus incipiens preserved pregnancies till term, but the specific glycoprotein was with in normal ranges. Spontaneous abortion occurred in 7% of women with low values under the 10th percentile. The present study show that examination of pregnancy specific beta 1-glycoprotein in women with threatened early pregnancy is of prognostic significance for the outcome of pregnancy.

  15. H-alpha observations of Sh2-190, Sh2-222, Sh2-229, Sh2-236 HII regions

    NASA Astrophysics Data System (ADS)

    Sahan, Muhittin

    2018-02-01

    Hα spectral line (6563Å) profiles of four northern HII regions in the our galaxy (Sh2-190, Sh2-222, Sh2-229, Sh2-236) have been obtained using DEFPOS spectrometer, located at coude focus of 150 cm RTT150 telescope at TUBITAK National Observatory (TUG, Antalya, Turkey). Observations were carried out at nights of 2015 December 24-27 with long exposure times ranging from 900s to 3600s. The LSR velocities and the linewidths (Full Width Half Maximum: FWHM) of the Hα emission lines were found to be in the range of -45.46 kms-1 to +3.57 kms-1 and 38.50 kms-1 to 44.10 kms-1, respectively. The Sh2-229 HII region is the faintest one (211.16 R), while the Sh2-236 HII region (IC410) is brightest source (535.75 R). The LSR velocity and the line width (FWHM) results of the DEFPOS/RTT150 system were compared with the data by several authors given in literature and results of DEFPOS data were found to be in good agreement with data given in literature.

  16. Area deprivation, individual socioeconomic status and low vision in the EPIC-Norfolk Eye Study.

    PubMed

    Yip, Jennifer L Y; Luben, Robert; Hayat, Shabina; Khawaja, Anthony P; Broadway, David C; Wareham, Nick; Khaw, K T; Foster, Paul J

    2014-03-01

    Poor vision is associated with lower socioeconomic status, but less is known about its relationship to area deprivation. The European Prospective Investigation into Cancer and Nutrition study Norfolk Eye Study was a cross-sectional study of 8563 participants with completed eye examinations. Logarithm of the minimum angle of resolution (logMAR) visual acuity (VA) was measured using standard protocols and low vision (LV) was defined as Snellen equivalent (VA) ≤6/12 in the better eye. Uncorrected refractive error (URE) was defined as improvement of VA by 2 logarithm of the minimum angle of resolution lines with pinhole. The lowest 5% of index of multiple deprivation rank was used to define the most deprived areas. The index of multiple deprivation is a composite measure using routine data from seven domains of deprivation to identify the most disadvantaged areas in England. Logistic regression was used to examine univariable and multivariable associations with LV. Ninety-six participants with missing data were excluded, leaving 8467 for analysis (98.9%). The mean age of the study group was 68.7 years (SD=8.1, range=48-92), with 55.1% women. LV was present in 263 participants (3.1%, 95% CI 2.7 to 3.5%). LV was associated with deprivation after adjusting for age, sex, education, social class and cataract surgery (OR=1.7, 95% CI 1.1 to 2.6, p=0.03), but this effect was mitigated by additionally adjusting for URE (OR=1.5, 95% CI 1.0 to 2.4, p=0.09). People with LV are more likely to live in the most deprived areas; this association was independent of socioeconomic status and partly mediated by URE. Targeting URE in deprived areas may reduce health inequalities associated with LV.

  17. Functionalized graphene oxide serves as a novel vaccine nano-adjuvant for robust stimulation of cellular immunity

    NASA Astrophysics Data System (ADS)

    Xu, Ligeng; Xiang, Jian; Liu, Ye; Xu, Jun; Luo, Yinchan; Feng, Liangzhu; Liu, Zhuang; Peng, Rui

    2016-02-01

    Benefiting from their unique physicochemical properties, graphene derivatives have attracted great attention in biomedicine. In this study, we carefully engineered graphene oxide (GO) as a vaccine adjuvant for immunotherapy using urease B (Ure B) as the model antigen. Ure B is a specific antigen for Helicobacter pylori, which is a class I carcinogen for gastric cancer. Polyethylene glycol (PEG) and various types of polyethylenimine (PEI) were used as coating polymers. Compared with single-polymer modified GOs (GO-PEG and GO-PEI), certain dual-polymer modified GOs (GO-PEG-PEI) can act as a positive modulator to promote the maturation of dendritic cells (DCs) and enhance their cytokine secretion through the activation of multiple toll-like receptor (TLR) pathways while showing low toxicity. Moreover, this GO-PEG-PEI can serve as an antigen carrier to effectively shuttle antigens into DCs. These two advantages enable GO-PEG-PEI to serve as a novel vaccine adjuvant. In the subsequent in vivo experiments, compared with free Ure B and clinically used aluminum-adjuvant-based vaccine (Alum-Ure B), GO-PEG-PEI-Ure B induces stronger cellular immunity via intradermal administration, suggesting promising applications in cancer immunotherapy. Our work not only presents a novel, highly effective GO-based vaccine nano-adjuvant, but also highlights the critical roles of surface chemistry for the rational design of nano-adjuvants.Benefiting from their unique physicochemical properties, graphene derivatives have attracted great attention in biomedicine. In this study, we carefully engineered graphene oxide (GO) as a vaccine adjuvant for immunotherapy using urease B (Ure B) as the model antigen. Ure B is a specific antigen for Helicobacter pylori, which is a class I carcinogen for gastric cancer. Polyethylene glycol (PEG) and various types of polyethylenimine (PEI) were used as coating polymers. Compared with single-polymer modified GOs (GO-PEG and GO-PEI), certain dual

  18. Regional Supplement to the Corps of Engineers Wetland Delineation Manual: Midwest Region (Version 2.0)

    DTIC Science & Technology

    2010-08-01

    2. Saplings and shrubs – 15-ft (4.6-m) radius 3. Herbaceous plants – 5-ft (1.5-m) radius or 3.28- by 3.28-ft square (1-m2) quadrat 4. Woody vines ...This supplement is applicable to the Midwest Region, which consists of all or portions of 14 states: Illinois, Indiana , Iowa, Kansas, Kentucky...This supplement is applicable to the Midwest Region, which consists of all or portions of 14 states: Illinois, Indiana , Iowa, Kansas, Kentucky

  19. Extracellular ammonia at sites of pulmonary infection with Coccidioides posadasii contributes to severity of the respiratory disease.

    PubMed

    Wise, Hua Zhang; Hung, Chiung-Yu; Whiston, Emily; Taylor, John W; Cole, Garry T

    2013-01-01

    Coccidioides is the causative agent of a potentially life-threatening respiratory disease of humans. A feature of this mycosis is that pH measurements of the microenvironment of pulmonary abscesses are consistently alkaline due to ammonia production during the parasitic cycle. We previously showed that enzymatically active urease is partly responsible for elevated concentrations of extracellular ammonia at sites of lung infection and contributes to both localized host tissue damage and exacerbation of the respiratory disease in BALB/c mice. Disruption of the urease gene (URE) of Coccidioides posadasii only partially reduced the amount of ammonia detected during in vitro growth of the parasitic phase, suggesting that other ammonia-producing pathways exist that may also contribute to the virulence of this pathogen. Ureidoglycolate hydrolase (Ugh) expressed by bacteria, fungi and higher plants catalyzes the hydrolysis of ureidoglycolate to yield glyoxylate and the release CO2 and ammonia. This enzymatic pathway is absent in mice and humans. Ureidoglycolate hydrolase gene deletions were conducted in a wild type (WT) isolate of C. posadasii as well as the previously generated Δure knock-out strain. Restorations of UGH in the mutant stains were performed to generate and evaluate the respective revertants. The double mutant revealed a marked decrease in the amount of extracellular ammonia without loss of reproductive competence in vitro compared to both the WT and Δure parental strains. BALB/c mice challenged intranasally with the Δugh/Δure mutant showed 90% survival after 30 days, decreased fungal burden, and well-organized pulmonary granulomas. We conclude that loss of both Ugh and Ure activity significantly reduced the virulence of this fungal pathogen. Copyright © 2013 Elsevier Ltd. All rights reserved.

  20. CO2 laser treatment for regional cutaneous malignant melanoma metastases.

    PubMed

    van Jarwaarde, Jorien A; Wessels, Ronnie; Nieweg, Omgo E; Wouters, Michel W J M; van der Hage, Jos A

    2015-01-01

    Cutaneous in-transit and satellite metastases are distressing presentations of melanoma progression. The purpose of this study was to analyze the efficacy of carbon dioxide (CO2) lasers in patients with melanoma with cutaneous in-transit and satellite metastases. Results of CO2 laser therapy were retrospectively evaluated in 22 patients between January 2004 and January 2008. The number of laser treatments, postoperative morbidity, regional control, and overall survival were analyzed. Twenty-two patients received a total of 42 CO2 laser treatments. The number of lesions treated per session varied from 3 to 329. The median duration of regional control in all patients was 14 weeks (range, 3-117). In 9 of 22 patients, only 1 treatment with CO2 laser was performed resulting in a mean regional control of 11 weeks. In 10 patients, an average of 4 laser treatments (range, 1-17) was necessary to achieve regional control. Three of the 22 patients underwent isolated limb perfusion after laser treatment for disease control. This study shows that (repeated) laser treatment can achieve adequate regional control with little morbidity. CO2 laser is recommended as a first-line treatment to patients with small but numerous cutaneous satellite or in-transit lesions in whom other surgery would induce substantial morbidity.

  1. Nonlinear regional warming with increasing CO2 concentrations

    NASA Astrophysics Data System (ADS)

    Good, Peter; Lowe, Jason A.; Andrews, Timothy; Wiltshire, Andrew; Chadwick, Robin; Ridley, Jeff K.; Menary, Matthew B.; Bouttes, Nathaelle; Dufresne, Jean Louis; Gregory, Jonathan M.; Schaller, Nathalie; Shiogama, Hideo

    2015-02-01

    When considering adaptation measures and global climate mitigation goals, stakeholders need regional-scale climate projections, including the range of plausible warming rates. To assist these stakeholders, it is important to understand whether some locations may see disproportionately high or low warming from additional forcing above targets such as 2 K (ref. ). There is a need to narrow uncertainty in this nonlinear warming, which requires understanding how climate changes as forcings increase from medium to high levels. However, quantifying and understanding regional nonlinear processes is challenging. Here we show that regional-scale warming can be strongly superlinear to successive CO2 doublings, using five different climate models. Ensemble-mean warming is superlinear over most land locations. Further, the inter-model spread tends to be amplified at higher forcing levels, as nonlinearities grow--especially when considering changes per kelvin of global warming. Regional nonlinearities in surface warming arise from nonlinearities in global-mean radiative balance, the Atlantic meridional overturning circulation, surface snow/ice cover and evapotranspiration. For robust adaptation and mitigation advice, therefore, potentially avoidable climate change (the difference between business-as-usual and mitigation scenarios) and unavoidable climate change (change under strong mitigation scenarios) may need different analysis methods.

  2. Global cost of correcting vision impairment from uncorrected refractive error.

    PubMed

    Fricke, T R; Holden, B A; Wilson, D A; Schlenther, G; Naidoo, K S; Resnikoff, S; Frick, K D

    2012-10-01

    To estimate the global cost of establishing and operating the educational and refractive care facilities required to provide care to all individuals who currently have vision impairment resulting from uncorrected refractive error (URE). The global cost of correcting URE was estimated using data on the population, the prevalence of URE and the number of existing refractive care practitioners in individual countries, the cost of establishing and operating educational programmes for practitioners and the cost of establishing and operating refractive care facilities. The assumptions made ensured that costs were not underestimated and an upper limit to the costs was derived using the most expensive extreme for each assumption. There were an estimated 158 million cases of distance vision impairment and 544 million cases of near vision impairment caused by URE worldwide in 2007. Approximately 47 000 additional full-time functional clinical refractionists and 18 000 ophthalmic dispensers would be required to provide refractive care services for these individuals. The global cost of educating the additional personnel and of establishing, maintaining and operating the refractive care facilities needed was estimated to be around 20 000 million United States dollars (US$) and the upper-limit cost was US$ 28 000 million. The estimated loss in global gross domestic product due to distance vision impairment caused by URE was US$ 202 000 million annually. The cost of establishing and operating the educational and refractive care facilities required to deal with vision impairment resulting from URE was a small proportion of the global loss in productivity associated with that vision impairment.

  3. Polymorphisms at the 3' untranslated region of SLC11A1 gene are associated with protection to Brucella infection in goats.

    PubMed

    Iacoboni, Paola A; Hasenauer, Flavia C; Caffaro, M Eugenia; Gaido, Analia; Rossetto, Cristina; Neumann, Roberto D; Salatin, Antonio; Bertoni, Emiliano; Poli, Mario A; Rossetti, Carlos A

    2014-08-15

    Goats are susceptible to brucellosis and the detection of Brucella-infected animals is carried out by serological tests. In other ruminant species, polymorphisms in microsatellites (Ms) of 3' untranslated region (3'UTR) of the solute carrier family 11 member A1 (SLC11A1) gene were associated with resistance to Brucella abortus infection. Goats present two polymorphic Ms at the 3'UTR end of SLC11A1 gene, called regions A and B. Here, we evaluated if polymorphisms in regions A and/or B are associated with Brucella infection in goats. Serum (for the detection of Brucella-specific antibodies) and hair samples (for DNA isolation and structure analysis of the SLC11A1 gene) were randomly collected from 229 adult native goats from the northwest of Argentina. Serological status was evaluated by buffer plate antigen test (BPAT) complemented by the fluorescent polarization assay (FPA), and the genotype of the 3'UTR of the SLC11A1 gene was determined by capillary electrophoresis and confirmed by sequence analysis. Polymorphisms in regions A and B of the 3'UTR SLC11A1 gene were found statistically significant associated with protection to Brucella infection. Specifically, the association study indicates statistical significance of the allele A15 and B7/B7 genotype with absence of Brucella-specific antibodies (p=0.0003 and 0.0088, respectively). These data open a promising opportunity for limiting goat brucellosis through selective breeding of animals based on genetic markers associated with natural resistance to B. melitensis infection. Copyright © 2014 Elsevier B.V. All rights reserved.

  4. Factors Influencing Student Gains from Undergraduate Research Experiences at a Hispanic-Serving Institution

    ERIC Educational Resources Information Center

    Daniels, Heather; Grineski, Sara E.; Collins, Timothy W.; Morales, Danielle X.; Morera, Osvaldo; Echegoyen, Lourdes

    2016-01-01

    Undergraduate research experiences (UREs) confer many benefits to students, including improved self-confidence, better communication skills, and an increased likelihood of pursuing science careers. Additionally, UREs may be particularly important for racial/ethnic minority students who are underrepresented in the science workforce. We examined…

  5. Ameliorative effects of gallic acid, quercetin and limonene on urethane-induced genotoxicity and oxidative stress in Drosophila melanogaster.

    PubMed

    Nagpal, Isha; Abraham, Suresh K

    2017-05-01

    The main objective of our present work was to ascertain the efficacy of Drosophila melanogaster model for assessing antigenotoxic and antioxidant effects of dietary phytochemicals gallic acid (GA), quercetin (QC) and limonene (Lim) against urethane (URE), a genotoxic environmental carcinogen. Oregon-K (ORK) adult male flies were fed GA, QC and Lim in combination with URE (20 mM) in 10% sucrose for 72 h. Third instar larvae were fed instant medium containing the above phytochemicals and URE for 24 h. Sex-linked recessive lethal (SLRL) test and assays for estimating glutathione content (GSH), glutathione S-transferase (GST), catalase (CAT), superoxide dismutase (SOD) and lipid peroxidation (MDA content) were performed. Adult feeding experiments demonstrated that co-treatment of flies with URE and the test phytochemicals has significantly decreased the frequencies of SLRL mutations in all the germ cell stages when compared to that with URE alone. Larval feeding experiments also showed a similar pattern. The above results correlate well with antioxidative potentials of the test agents where we observed the elevated enzymatic levels with a significant reduction in MDA level in Drosophila larvae. The results further suggest that the dietary phytochemicals have an antioxidant and antimutagenic property which can be assessed using D. melanogaster.

  6. Genetic diversity of clinical and environmental Vibrio parahaemolyticus strains from the Pacific Northwest.

    PubMed

    Paranjpye, Rohinee; Hamel, Owen S; Stojanovski, Asta; Liermann, Martin

    2012-12-01

    Since 1997, cases of Vibrio parahaemolyticus-related gastroenteritis from the consumption of raw oysters harvested in Washington State have been higher than historical levels. These cases have shown little or no correlation with concentrations of potentially pathogenic V. parahaemolyticus (positive for the thermostable direct hemolysin gene, tdh) in oysters, although significant concentrations of tdh(+) V. parahaemolyticus strains were isolated from shellfish-growing areas in the Pacific Northwest (PNW). We compared clinical and environmental strains isolated from the PNW to those from other geographic regions within the United States and Asia for the presence of virulence-associated genes, including the thermostable direct hemolysin (tdh), the thermostable-related hemolysin (trh), urease (ureR), the pandemic group specific markers orf8 and toxRS, and genes encoding both type 3 secretion systems (T3SS1 and T3SS2). The majority of clinical strains from the PNW were positive for tdh, trh, and ureR genes, while a significant proportion of environmental isolates were tdh(+) but trh negative. Hierarchical clustering grouped the majority of these clinical isolates into a cluster distinct from that including the pandemic strain RIMD2210633, clinical isolates from other geographical regions, and tdh(+), trh-negative environmental isolates from the PNW. We detected T3SS2-related genes (T3SS2β) in environmental strains that were tdh and trh negative. The presence of significant concentrations of tdh(+), trh-negative environmental strains in the PNW that have not been responsible for illness and T3SS2β in tdh- and trh-negative strains emphasizes the diversity in this species and the need to identify additional virulence markers for this bacterium to improve risk assessment tools for the detection of this pathogen.

  7. Telecommunications forecast for ITU Region 2 to the year 1995

    NASA Technical Reports Server (NTRS)

    Hollansworth, J. E.; Salzman, J. A.; Ramler, J. R.

    1985-01-01

    Telecommunications activity was studied. The primary objective was to forecast the need for fixed service satellites (FSS) by countries within ITU Region 2 excluding the United States and Greenland. Forecasts of telecommunications equipment needs were developed as a yardstick of the relative level of telecommunications activity among developing countries within the region. A likely scenario for the implementation of domestic and regional communications satellites is forecasted to provide services to and among countries in ITU Region 2. By 1995, it is forecast that 15 fixed service satellites will be implemented. A forecast of the countries requirements indicates that, with the possible exception of Canada, this constellation of satellites will meet these countries' needs to beyond the year 2000.

  8. Wide band antireflective coatings Al2O3 / HfO2 / MgF2 for UV region

    NASA Astrophysics Data System (ADS)

    Winkowski, P.; Marszałek, Konstanty W.

    2013-07-01

    Deposition technology of the three layers antireflective coatings consists of hafnium compound are presented in this paper. Oxide films were deposited by means of e-gun evaporation in vacuum of 5x10-5 mbar in presence of oxygen and fluoride films by thermal evaporation. Substrate temperature was 250°C. Coatings were deposited onto optical lenses made from quartz glass (Corning HPFS). Thickness and deposition rate were controlled by thickness measuring system Inficon XTC/2. Simulations leading to optimization of thickness and experimental results of optical measurements carried during and after deposition process were presented. Physical thickness measurements were made during deposition process and were equal to 43 nm/74 nm/51 nm for Al2O3 / HfO2 / MgF2 respectively. Optimization was carried out for ultraviolet region from 230nm to the beginning of visible region 400 nm. In this region the average reflectance of the antireflective coating was less than 0.5% in the whole range of application.

  9. Experimental infection of nontarget species of rodents and birds with Brucella abortus strain RB51 vaccine

    USGS Publications Warehouse

    Januszewski, M.C.; Olsen, S.C.; McLean, R.G.; Clark, L.; Rhyan, Jack C.

    2001-01-01

    The Brucella abortus vaccine strain RB51 (SRB51) is being considered for use in the management of bnucellosis in wild bison (Bison bison) and elk (Cervus elaphus) populations in the Greater Yellowstone Area (USA). Evaluation of the vaccines safety in non-target species was considered necessary prior to field use. Between June 1998 and December 1999, ground squirrels (Spermophilus richardsonii, n = 21), deer mice (Peromyscus maniculatus, n = 14), prairie voles (Microtus ochrogaster, n = 21), and ravens (Corvus corax, n = 13) were orally inoculated with SRB51 or physiologic saline. Oral and rectal swabs and blood samples were collected for bacteriologic evaluation. Rodents were necropsied at 8 to 10 wk and 12 to 21 wk post inoculation (PI), and ravens at 7 and 11 wk PI. Spleen, liver and reproductive tissues were collected for bacteriologic and histopathologic evaluation. No differences in clinical signs, appetite, weight loss or gain, or activity were observed between saline- and SRB51-inoculated animals in all four species. Oral and rectal swabs from all species were negative throughout the study. In tissues obtained from SRB51-inoculated animals, the organism was isolated from six of seven (86%) ground squirrels, one of six (17%) deer mice, none of seven voles, and one of five (20%) ravens necropsied at 8, 8, 10, and 7 wk PI, respectively. Tissues from four of seven (57%) SRB51-inoculated ground squirrels were culture positive for the organism 12 wk PI; SRB51 was not recovered from deer mice, voles. or ravens necropsied 12, 21, or 11 wk, respectively, PI. SRB51 was not recovered from saline-inoculated ground squirrels, deer mice, or voles at any time but was recovered from one saline-inoculated raven at necropsy, 7 wk PI, likely attributable to contact with SRB51-inoculated ravens in an adjacent aviary room. Spleen was time primary tissue site of colonization in ground squirrels, followed by the liver and reproductive organs. The results indicate oral exposure to

  10. Construction of two ureolytic model organisms for the study of microbially induced calcium carbonate precipitation.

    PubMed

    Connolly, James; Kaufman, Megan; Rothman, Adam; Gupta, Rashmi; Redden, George; Schuster, Martin; Colwell, Frederick; Gerlach, Robin

    2013-09-01

    Two bacterial strains, Pseudomonas aeruginosa MJK1 and Escherichia coli MJK2, were constructed that both express green fluorescent protein (GFP) and carry out ureolysis. These two novel model organisms are useful for studying bacterial carbonate mineral precipitation processes and specifically ureolysis-driven microbially induced calcium carbonate precipitation (MICP). The strains were constructed by adding plasmid-borne urease genes (ureABC, ureD and ureFG) to the strains P. aeruginosa AH298 and E. coli AF504gfp, both of which already carried unstable GFP derivatives. The ureolytic activities of the two new strains were compared to the common, non-GFP expressing, model organism Sporosarcina pasteurii in planktonic culture under standard laboratory growth conditions. It was found that the engineered strains exhibited a lower ureolysis rate per cell but were able to grow faster and to a higher population density under the conditions of this study. Both engineered strains were successfully grown as biofilms in capillary flow cell reactors and ureolysis-induced calcium carbonate mineral precipitation was observed microscopically. The undisturbed spatiotemporal distribution of biomass and calcium carbonate minerals were successfully resolved in 3D using confocal laser scanning microscopy. Observations of this nature were not possible previously because no obligate urease producer that expresses GFP had been available. Future observations using these organisms will allow researchers to further improve engineered application of MICP as well as study natural mineralization processes in model systems. © 2013.

  11. Myelodysplastic syndromes and acute myeloid leukemia in cats infected with feline leukemia virus clone33 containing a unique long terminal repeat.

    PubMed

    Hisasue, Masaharu; Nagashima, Naho; Nishigaki, Kazuo; Fukuzawa, Isao; Ura, Shigeyoshi; Katae, Hiromi; Tsuchiya, Ryo; Yamada, Takatsugu; Hasegawa, Atsuhiko; Tsujimoto, Hajime

    2009-03-01

    Feline leukemia virus (FeLV) clone33 was obtained from a domestic cat with acute myeloid leukemia (AML). The long terminal repeat (LTR) of this virus, like the LTRs present in FeLV from other cats with AML, differs from the LTRs of other known FeLV in that it has 3 tandem direct 47-bp repeats in the upstream region of the enhancer (URE). Here, we injected cats with FeLV clone33 and found 41% developed myelodysplastic syndromes (MDS) characterized by peripheral blood cytopenias and dysplastic changes in the bone marrow. Some of the cats with MDS eventually developed AML. The bone marrow of the majority of cats with FeLV clone33 induced MDS produced fewer erythroid and myeloid colonies upon being cultured with erythropoietin or granulocyte-macrophage colony-stimulating factor (GM-SCF) than bone marrow from normal control cats. Furthermore, the bone marrow of some of the cats expressed high-levels of the apoptosis-related genes TNF-alpha and survivin. Analysis of the proviral sequences obtained from 13 cats with naturally occurring MDS reveal they also bear the characteristic URE repeats seen in the LTR of FeLV clone33 and other proviruses from cats with AML. Deletions and mutations within the enhancer elements are frequently observed in naturally occurring MDS as well as AML. These results suggest that FeLV variants that bear URE repeats in their LTR strongly associate with the induction of both MDS and AML in cats.

  12. Actinobacillus pleuropneumoniae Iron Transport and Urease Activity: Effects on Bacterial Virulence and Host Immune Response

    PubMed Central

    Baltes, Nina; Tonpitak, Walaiporn; Gerlach, Gerald-F.; Hennig-Pauka, Isabel; Hoffmann-Moujahid, Astrid; Ganter, Martin; Rothkötter, Hermann-J.

    2001-01-01

    Actinobacillus pleuropneumoniae, a porcine respiratory tract pathogen, has been shown to express transferrin-binding proteins and urease during infection. Both activities have been associated with virulence; however, their functional role for infection has not yet been elucidated. We used two isogenic A. pleuropneumoniae single mutants (ΔexbB and ΔureC) and a newly constructed A. pleuropneumoniae double (ΔureC ΔexbB) mutant in aerosol infection experiments. Neither the A. pleuropneumoniae ΔexbB mutant nor the double ΔureC ΔexbB mutant was able to colonize sufficiently long to initiate a detectable humoral immune response. These results imply that the ability to utilize transferrin-bound iron is required for multiplication and persistence of A. pleuropneumoniae in the porcine respiratory tract. The A. pleuropneumoniae ΔureC mutant and the parent strain both caused infections that were indistinguishable from one another in the acute phase of disease; however, 3 weeks postinfection the A. pleuropneumoniae ΔureC mutant, in contrast to the parent strain, could not be isolated from healthy lung tissue. In addition, the local immune response—as assessed by fluorescence-activated cell sorter and enzyme-linked immunosorbent spot analyses—revealed a significantly higher number of A. pleuropneumoniae-specific B cells in the bronchoalveolar lavage fluid (BALF) of pigs infected with the A. pleuropneumoniae ΔureC mutant than in the BALF of those infected with the parent strain. These results imply that A. pleuropneumoniae urease activity may cause sufficient impairment of the local immune response to slightly improve the persistence of the urease-positive A. pleuropneumoniae parent strain. PMID:11119539

  13. Molecular dynamics simulations of Hsp40 J-domain mutants identifies disruption of the critical HPD-motif as the key factor for impaired curing in vivo of the yeast prion [URE3].

    PubMed

    Xue, You-Lin; Wang, Hao; Riedy, Michael; Roberts, Brittany-Lee; Sun, Yuna; Song, Yong-Bo; Jones, Gary W; Masison, Daniel C; Song, Youtao

    2018-05-01

    Genetic screens using Saccharomyces cerevisiae have identified an array of Hsp40 (Ydj1p) J-domain mutants that are impaired in the ability to cure the yeast [URE3] prion through disrupting functional interactions with Hsp70. However, biochemical analysis of some of these Hsp40 J-domain mutants has so far failed to provide major insight into the specific functional changes in Hsp40-Hsp70 interactions. To explore the detailed structural and dynamic properties of the Hsp40 J-domain, 20 ns molecular dynamic simulations of 4 mutants (D9A, D36A, A30T, and F45S) and wild-type J-domain were performed, followed by Hsp70 docking simulations. Results demonstrated that although the Hsp70 interaction mechanism of the mutants may vary, the major structural change was targeted to the critical HPD motif of the J-domain. Our computational analysis fits well with previous yeast genetics studies regarding highlighting the importance of J-domain function in prion propagation. During the molecular dynamics simulations several important residues were identified and predicted to play an essential role in J-domain structure. Among these residues, Y26 and F45 were confirmed, using both in silico and in vivo methods, as being critical for Ydj1p function.

  14. Seroprevalence and risk factors associated with brucellosis as a professional hazard in Pakistan.

    PubMed

    Ali, Shahzad; Ali, Qurban; Neubauer, Heinrich; Melzer, Falk; Elschner, Mandy; Khan, Iahtasham; Abatih, Emmanuel Nji; Ullah, Nemat; Irfan, Muhammad; Akhter, Shamim

    2013-06-01

    The present study was conducted to determine the seroprevalence and identify risk factors associated with brucellosis in humans at high risk in the Potohar plateau of northeastern Pakistan. A total of 262 serum samples were collected from persons of different occupational groups: veterinary personnel, milkers, abattoir workers, livestock farmers, and others (drivers, security guards, housewives). Data related to gender, age, occupation, contact with animals, brucellosis-related symptoms, consumption of raw milk, and geographical region were collected. The Rose Bengal plate test and the serum agglutination test were performed to determine the seroprevalence of brucellosis. The overall seroprevalence was found to be 6.9% (95% confidence interval [CI]: 4.1, 10.6). Real-time polymerase chain reaction assay showed that all cases were affected by Brucella abortus. Individuals who consumed raw milk had higher odds of brucellosis seropositivity. This is the first report of human brucellosis related to B. abortus in high-risk professionals from Pakistan by the combined use of serological and molecular methods.

  15. Cultivating Minority Scientists: Undergraduate Research Increases Self-Efficacy and Career Ambitions for Underrepresented Students in STEM

    ERIC Educational Resources Information Center

    Carpi, Anthony; Ronan, Darcy M.; Falconer, Heather M.; Lents, Nathan H.

    2017-01-01

    In this study, Social Cognitive Career Theory (SCCT) is used to explore changes in the career intentions of students in an undergraduate research experience (URE) program at a large public minority-serving college. Our URE model addresses the challenges of establishing an undergraduate research program within an urban, commuter, underfunded,…

  16. NGA-West2 Empirical Fourier Model for Active Crustal Regions to Generate Regionally Adjustable Response Spectra

    NASA Astrophysics Data System (ADS)

    Bora, S. S.; Cotton, F.; Scherbaum, F.; Kuehn, N. M.

    2016-12-01

    Adjustment of median ground motion prediction equations (GMPEs) from data-rich (host) regions to data-poor regions (target) is one of major challenges that remains with the current practice of engineering seismology and seismic hazard analysis. Fourier spectral representation of ground motion provides a solution to address the problem of adjustment that is physically transparent and consistent with the concepts of linear system theory. Also, it provides a direct interface to appreciate the physically expected behavior of seismological parameters on ground motion. In the present study, we derive an empirical Fourier model for computing regionally adjustable response spectral ordinates based on random vibration theory (RVT) from shallow crustal earthquakes in active tectonic regions, following the approach of Bora et al. (2014, 2015). , For this purpose, we use an expanded NGA-West2 database with M 3.2—7.9 earthquakes at distances ranging from 0 to 300 km. A mixed-effects regression technique is employed to further explore various components of variability. The NGA-West2 database expanded over a wide magnitude range provides a better understanding (and constraint) of source scaling of ground motion. The large global volume of the database also allows investigating regional patterns in distance-dependent attenuation (i.e., geometrical spreading and inelastic attenuation) of ground motion as well as in the source parameters (e.g., magnitude and stress drop). Furthermore, event-wise variability and its correlation with stress parameter are investigated. Finally, application of the derived Fourier model in generating adjustable response spectra will be shown.

  17. Petroleum exploration plays and resource estimates, 1989, onshore United States; Region 1, Alaska; Region 2, Pacific Coast

    USGS Publications Warehouse

    Powers, Richard B.

    1993-01-01

    This study provides brief discussions of the petroleum geology, play descriptions, and resource estimates of 220 individually assessed exploration plays in all 80 onshore geologic provinces within nine assessment regions of the continental United States in 1989; these 80 onshore provinces were assessed in connection with the determination of the Nation's estimated undiscovered resources of oil and gas. The present report covers the 25 provinces that make up Region 1, Alaska, and Region 2, Pacific Coast. It is our intention to issue Region 3, Colorado Plateau and Basin and Range, and Region 4, Rocky Mountains and Northern Great Plains, in book form as well. Regions 5 through 9 (West Texas and Eastern New Mexico, Gulf Coast, Midcontinent, Eastern Interior and Atlantic Coast) will be released individually, as Open-File Reports.

  18. The effect of secular resonances in the asteroid region between 2.1 and 2.4 AU

    NASA Technical Reports Server (NTRS)

    Froeschle, Christiane; Scholl, Hans

    1992-01-01

    The asteroid region between 2.1 and 2.4 AU appears to be depopulated at inclinations i greater than 12 deg. This region is surrounded by the three main secular resonances nu(sub 5), nu(sub 6), and nu(sub 16) and is crossed by higher order secular resonances. Secular resonances appear to overlap in this region. Numerical integrations of the orbits of seventeen fictituous asteroids with initial inclinations 12 deg less than or equal to i less than or equal to 20 deg show the following: (1) this particular asteroid region is not depopulated in our computer experiment on timescales of 2.7 Myrs; (2) inclinations are pumped up by successive crossings through higher order secular resonances while eccentricities are not increased sufficiently to produce planet-crossers; (3) bodies located in the bordering nu(sub 6) resonance with semi-major axes a less than or equal to 2.4 AU become Earth-crossers on a time scale of 1 Myr; and (4) we confirm the result that modes due to higher order secular resonances must be eliminated when proper elements are computed.

  19. Analysis of Student Performance on the Undergraduate Record Examinations (1973).

    ERIC Educational Resources Information Center

    Litwin, James L.

    This report examines the performance of students in the Modular Achievement Program (MAP) at Bowling Green State University using the Undergraduate Record Examinations (URE) as the primary criterion. The performances of students in MAP on the URE is delineated and compared to the performance of freshman and sophomore norm groups at Bowling Green…

  20. Evaluation of a BSW Research Experience: Improving Student Research Competency

    ERIC Educational Resources Information Center

    Whipple, Ellen E.; Hughes, Anne; Bowden, Susan

    2015-01-01

    This article examines the experience of 24 BSW students in a faculty-mentored undergraduate research experience (URE) over the course of 1 academic year. In particular, we sought to better understand students' self-perceived sense of competency across 15 specific research skills. In addition, we examined the URE's impact on students' knowledge…

  1. Testing the Impact of a Multi-year, Curriculum-based Undergraduate Research Experience (MY-CURE) in the Geosciences: Baseline Observations

    NASA Astrophysics Data System (ADS)

    Allen, J. L.; Creamer, E. G.; Kuehn, S. C.

    2016-12-01

    Short-term undergraduate research experiences (URE's) provide skill and confidence enhancement to students, but it is unclear how effective they are in comparison to a dedicated, longer-term URE. This study examines the impact of a long-term URE embedded in a sequence of five courses in the geology curriculum. It begins with a sophomore course in environmental geology, and continues through mineralogy, structural geology, and petrology, before concluding at our summer geology field camp. In this sequence, they build upon individual URE's related to the structure and petrology of fault rocks from a mid-crustal shear zone. Rather than have students engage in one or more short-term URE's, they retain the same project for two calendar years so that we can assess when and how different gains, including a more sophisticated understanding of the nature of science, begin to emerge and mature. As each student progresses, we document the longitudinal development of a diverse suite of gains including: (1) Technical and higher-order research skills, (2) personal gains such as self-identity as a scientist, and (3) communication skills. In this presentation, we describe the framework of the study and baseline observations recorded during the first year of a 2-year cohort. Using a Q-sort method, students were given a deck of 16 index cards with an educational outcome listed on each. They sorted the cards into three piles: Those that encouraged an interest in geology, those that deterred an interest, and those with no impact. Participants discussed the top cards from the negative and positive piles. The top attractors to geology are collegial relationships with faculty, the opportunity to use scientific equipment, field work, the concreteness of geology, and the availability of jobs. Factors that deter interest include hours of tedious homework, math courses, and time invested in wrong answers or failed experiments/sample preparation. Factors not yet evident include confidence in

  2. A new Tla region antigen Qa-11, similar to Qa-2 and associated with B-type beta 2-microglobulin.

    PubMed

    van de Meugheuvel, W; van Seventer, G; Demant, P

    1985-04-01

    A new antigen, Qa-11, is detected as a 40,000 dalton band in the SDS-PAGE of immunoprecipitates of radiolabeled lymphocyte membrane preparations. In C57BL H-2 congenic strains, its presence is controlled by a gene in the Tla region. In strains with genetic background other than C57BL it is not expressed. Tests with recombinant inbred strains and with H-3 congenic strains show that, in addition to the Tla region, a gene linked to or identical with the beta 2-microglobulin-b-allele is required for the expression of Qa-11 as well. The mobility of the Qa-11 antigen in SDS-PAGE and in isoelectrofocusing is the same as that of Qa-2 antigen. The Cleveland peptide maps of Qa-2 and Qa-11 are identical as well. This finding, that the Tla region controlled Qa-11 antigen is structurally very similar to the Qa-2 antigen, contrasts with the fact that Tla region products do not react with anti-Qa-2 sera. This paradox could be explained by a separate Qa-11 region between Qa-2 and Tla. Alternatively, it is possible that the Qa-11 antigen is the result of the action of a modifying gene in the Tla region upon a Qa-2 gene product, or that the structural gene for Qa-11 is located in the Qa-2 region and a Tla region gene controls its expression.

  3. Region 2 Port-area Investigation of Emissions Reduction (R2PIER)

    EPA Science Inventory

    Background Region 2 is home to the Port of New York and New Jersey (Port), the largest marine port on the East Coast and third largest in the nation. The Port is a concentrated source of diesel pollution, as more than 3 million containers move each year on diesel-powered ships, ...

  4. 47 CFR Appendix 2 to Part 97 - VEC Regions

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... SERVICE Pt. 97, App. 2 Appendix 2 to Part 97—VEC Regions 1. Connecticut, Maine, Massachusetts, New Hampshire, Rhode Island and Vermont. 2. New Jersey and New York. 3. Delaware, District of Columbia, Maryland... and Virginia. 5. Arkansas, Louisiana, Mississippi, New Mexico, Oklahoma and Texas. 6. California. 7...

  5. Discrimination of Enterohemorrhagic Escherichia coli (EHEC) from Non-EHEC Strains Based on Detection of Various Combinations of Type III Effector Genes

    PubMed Central

    Delannoy, Sabine; Beutin, Lothar

    2013-01-01

    Enterohemorrhagic Escherichia coli (EHEC) strains comprise a subgroup of Shiga-toxin (Stx)-producing E. coli (STEC) and are characterized by a few serotypes. Among these, seven priority STEC serotypes (O26:H11, O45:H2, O103:H2, O111:H8, O121:H19, O145:H28, and O157:H7) are most frequently implicated in severe clinical illness worldwide. Currently, standard methods using stx, eae, and O-serogroup-specific gene sequences for detecting the top 7 EHEC serotypes bear the disadvantage that these genes can be found in non-EHEC strains as well. Here, we explored the suitability of ureD, espV, espK, espN, Z2098, and espM1 genes and combinations thereof as candidates for a more targeted EHEC screening assay. For a very large panel of E. coli strains (n = 1,100), which comprised EHEC (n = 340), enteropathogenic E. coli (EPEC) (n = 392), STEC (n = 193), and apathogenic strains (n = 175), we showed that these genetic markers were more prevalent in EHEC (67.1% to 92.4%) than in EPEC (13.3% to 45.2%), STEC (0.5% to 3.6%), and apathogenic E. coli strains (0 to 2.9%). It is noteworthy that 38.5% of the EPEC strains that tested positive for at least one of these genetic markers belonged to the top 7 EHEC serotypes, suggesting that such isolates might be Stx-negative derivatives of EHEC. The associations of espK with either espV, ureD, or Z2098 were the best combinations for more specific and sensitive detection of the top 7 EHEC strains, allowing detection of 99.3% to 100% of these strains. In addition, detection of 93.7% of the EHEC strains belonging to other serotypes than the top 7 offers a possibility for identifying new emerging EHEC strains. PMID:23884997

  6. Frictional Heating of Ions In The F2-region of The Ionosphere

    NASA Astrophysics Data System (ADS)

    Zhizhko, G. O.; Vlasov, V. G.

    Auroral electron beams unstable on the Cherenkov resonance are stabilized by large- scale inhomogeneity of the plasma density during all their way from the acceleration region to the E-region of the ionosphere. The generation of plasma waves by beam is possible only in the region of small plasma density gradients, that always is the area of the F2-region maximum. Thus, collective dissipation of the electron beam energy occurs in the local region with the length about several tens of kilometers. This leads to the intensive heating of the electrons(up to temperatures about 10000 K) and will give origin to the ion upflows with velocity about 1 km/s and density about 109 cm-2 s-1. These flows can result in the ion frictional heating. At the same time ion temperatures reach the values about 5000 K. A numerical simulation of the ion frictional heating in the presence of collective elec- tron heating in the high-latitude F2-region of the ionosphere was performed. The sim- ulation has shown that the most critical parameter for the occurence of the ion fric- tional heating was the the steepness of the plasma density profile above the F2-region maximum.

  7. Independent activity of the homologous small regulatory RNAs AbcR1 and AbcR2 in the legume symbiont Sinorhizobium meliloti.

    PubMed

    Torres-Quesada, Omar; Millán, Vicenta; Nisa-Martínez, Rafael; Bardou, Florian; Crespi, Martín; Toro, Nicolás; Jiménez-Zurdo, José I

    2013-01-01

    The legume symbiont Sinorhizobium meliloti expresses a plethora of small noncoding RNAs (sRNAs) whose function is mostly unknown. Here, we have functionally characterized two tandemly encoded S. meliloti Rm1021 sRNAs that are similar in sequence and structure. Homologous sRNAs (designated AbcR1 and AbcR2) have been shown to regulate several ABC transporters in the related α-proteobacteria Agrobacterium tumefaciens and Brucella abortus. In Rm1021, AbcR1 and AbcR2 exhibit divergent unlinked regulation and are stabilized by the RNA chaperone Hfq. AbcR1 is transcribed in actively dividing bacteria, either in culture, rhizosphere or within the invasion zone of mature alfalfa nodules. Conversely, AbcR2 expression is induced upon entry into stationary phase and under abiotic stress. Only deletion of AbcR1 resulted into a discrete growth delay in rich medium, but both are dispensable for symbiosis. Periplasmic proteome profiling revealed down-regulation of the branched-chain amino acid binding protein LivK by AbcR1, but not by AbcR2. A double-plasmid reporter assay confirmed the predicted specific targeting of the 5'-untranslated region of the livK mRNA by AbcR1 in vivo. Our findings provide evidences of independent regulatory functions of these sRNAs, probably to fine-tune nutrient uptake in free-living and undifferentiated symbiotic rhizobia.

  8. Independent Activity of the Homologous Small Regulatory RNAs AbcR1 and AbcR2 in the Legume Symbiont Sinorhizobium meliloti

    PubMed Central

    Torres-Quesada, Omar; Millán, Vicenta; Nisa-Martínez, Rafael; Bardou, Florian; Crespi, Martín; Toro, Nicolás; Jiménez-Zurdo, José I.

    2013-01-01

    The legume symbiont Sinorhizobium meliloti expresses a plethora of small noncoding RNAs (sRNAs) whose function is mostly unknown. Here, we have functionally characterized two tandemly encoded S. meliloti Rm1021 sRNAs that are similar in sequence and structure. Homologous sRNAs (designated AbcR1 and AbcR2) have been shown to regulate several ABC transporters in the related α-proteobacteria Agrobacterium tumefaciens and Brucella abortus. In Rm1021, AbcR1 and AbcR2 exhibit divergent unlinked regulation and are stabilized by the RNA chaperone Hfq. AbcR1 is transcribed in actively dividing bacteria, either in culture, rhizosphere or within the invasion zone of mature alfalfa nodules. Conversely, AbcR2 expression is induced upon entry into stationary phase and under abiotic stress. Only deletion of AbcR1 resulted into a discrete growth delay in rich medium, but both are dispensable for symbiosis. Periplasmic proteome profiling revealed down-regulation of the branched-chain amino acid binding protein LivK by AbcR1, but not by AbcR2. A double-plasmid reporter assay confirmed the predicted specific targeting of the 5′-untranslated region of the livK mRNA by AbcR1 in vivo. Our findings provide evidences of independent regulatory functions of these sRNAs, probably to fine-tune nutrient uptake in free-living and undifferentiated symbiotic rhizobia. PMID:23869210

  9. Vane array with one or more non-integral platforms

    DOEpatents

    Lohaus, Andrew S.; Campbell, Christian Xavier; Miller, Jr, Samuel R.; Marra, John J.

    2016-07-12

    A vane array adapted to be coupled to a vane carrier within a gas turbine engine is provided comprising: a plurality of elongated airfoils comprising at least a first airfoil and a second airfoil located adjacent to one another; a U-ring; first connector structure for coupling a radially inner end section of each of the first and second airfoils to the U-ring; second connector structure for coupling a radially outer end section of each of the first and second airfoils to the vane carrier; a platform extending between the first and second airfoils; and platform connector structure for coupling the platform to one of the U-ring and the vane carrier.

  10. Combined deficiency of MSH2 and Sμ region abolishes class switch recombination.

    PubMed

    Leduc, Claire; Haddad, Dania; Laviolette-Malirat, Nathalie; Nguyen Huu, Ngoc-Sa; Khamlichi, Ahmed Amine

    2010-10-01

    Class switch recombination (CSR) is mediated by G-rich tandem repeated sequences termed switch regions. Transcription of switch regions generates single-stranded R loops that provide substrates for activation-induced cytidine deaminase. Mice deficient in MSH2 have a mild defect in CSR and analysis of their switch junctions has led to a model in which MSH2 is more critical for switch recombination events outside than within the tandem repeats. It is also known that deletion of the whole Sμ region severely impairs but does not abrogate CSR despite the lack of detectable R loops. Here, we demonstrate that deficiency of both MSH2 and the Sμ region completely abolishes CSR and that the abrogation occurs at the genomic level. This finding further supports the crucial role of MSH2 outside the tandem repeats. It also indicates that during CSR, MSH2 has access to activation-induced cytidine deaminase targets in R-loop-deficient Iμ-Cμ sequences rarely used in CSR, suggesting an MSH2-dependent DNA processing activity at the Iμ exon that may decrease with transcription elongation across the Sμ region.

  11. Brucellosis Transmission between Wildlife and Livestock in the Greater Yellowstone Ecosystem: Inferences from DNA Genotyping.

    PubMed

    O'Brien, Michael P; Beja-Pereira, Albano; Anderson, Neil; Ceballos, Ruben M; Edwards, William H; Harris, Beth; Wallen, Rick L; Costa, Vânia

    2017-04-01

    The wildlife of the Greater Yellowstone Ecosystem carries brucellosis, which was first introduced to the area by cattle in the 19th century. Brucellosis transmission between wildlife and livestock has been difficult to study due to challenges in culturing the causative agent, Brucella abortus . We examined B. abortus transmission between American bison ( Bison bison ), Rocky Mountain elk ( Cervus elaphus nelsoni), and cattle ( Bos taurus ) using variable number tandem repeat (VNTR) markers on DNA from 98 B. abortus isolates recovered from populations in Idaho, Montana, and Wyoming, US. Our analyses reveal interspecies transmission. Two outbreaks (2007, 2008) in Montana cattle had B. abortus genotypes similar to isolates from both bison and elk. Nevertheless, similarity in elk and cattle isolates from the 2008 outbreak suggest that elk are the likely source of brucellosis transmission to cattle in Montana and Wyoming. Brucella abortus isolates from sampling in Montana appear to be divided in two clusters: one found in local Montana elk, cattle, and bison; and another found mainly in elk and a bison from Wyoming, which is consistent with brucellosis having entered Montana via migration of infected elk from Wyoming. Our findings illustrate complex patterns of brucellosis transmission among elk, bison, and cattle as well as the utility of VNTRs to infer the wildlife species of origin for disease outbreaks in livestock.

  12. EPA Region 2 SEMS_CERCLIS Sites All [R2] and SEMS_CERCLIS Sites NPL [R2] GIS Layers

    EPA Pesticide Factsheets

    The Region 2 SEMS_CERCLIS Sites All [R2] GIS layer contains unique Superfund Enterprise Management System (SEMS) site records. These records have the following NPL_STATUS designations: CURRENTLY ON FINAL NPL, DELETED FROM FINAL NPL, NOT ON NPL, PROPOSED FOR NPL, REMOVED FROM PROPOSED NPL, and SITE IS PART OF NPL SITE. The Region 2 SEMS_CERCLIS NPL Sites [R2] GIS layer only has SEMS records with the following NPL_STATUS designations: 'CURRENTLY ON FINAL NPL', 'DELETED FROM FINAL NPL', 'PROPOSED FOR NPL'.The Superfund Enterprise Management System (SEMS) is EPA's official record for tracking hazardous waste sites, potentially hazardous waste sites, and remedial activities performed in support of the Superfund Program across the nation. This includes sites that are on the National Priorities List (NPL) or are being considered for the NPL. SEMS represents a joint development and ongoing collaboration between Superfund's Remedial, Removal, Federal Facilities, Enforcement, and Emergency Response programs. It provides its wide audience base with a means of ongoing analysis of Superfund Program activities and informational needs at the site, regional management, and national management levels. The customers of SEMS or SEMS data are five EPA Headquarters offices and regional staff, citizens, the regulated community, other Federal agencies, States, Tribes, local agencies, and industry. SEMS stakeholders are States, Congress, other Federal agencies, industry groups, and cit

  13. Vaccination with recombinant L7/L12-truncated Omp31 protein induces protection against Brucella infection in BALB/c mice.

    PubMed

    Golshani, Maryam; Rafati, Sima; Dashti, Amir; Gholami, Elham; Siadat, Seyed Davar; Oloomi, Mana; Jafari, Anis; Bouzari, Saeid

    2015-06-01

    Brucellosis is the most common bacterial zoonotic disease worldwide and no vaccine is available for the prevention of human brucellosis. In humans, brucellosis is mostly caused by Brucella melitensis and Brucella abortus. The Outer membrane protein 31 (Omp31) and L7/L12 are immunodominant and protective antigens conserved in human Brucella pathogens. In the present study, we evaluated the humoral and cellular immune responses induced by a fusion protein designed based on the Truncated form of Omp31 (TOmp31) and L7-L12 antigens. Vaccination of BALB/c mice with the recombinant fusion protein (rL7/L12-TOmp31) provided the significant protection level against B. melitensis and B. abortus challenge. Moreover, rL7/L12-TOmp31 elicited a strong specific IgG response (higher IgG2a titers) and significant IFN-γ/IL2 production and T-cell proliferation was also observed. The T helper1 (Th1) oriented response persisted for 12 weeks after the first immunization. The rL7/L12-TOmp31 could be a new potential antigen candidate for the development of a subunit vaccine against B. melitensis and B. abortus. Copyright © 2015 Elsevier Ltd. All rights reserved.

  14. 2-D Structure of the A Region of Xist RNA and Its Implication for PRC2 Association

    PubMed Central

    Maenner, Sylvain; Blaud, Magali; Fouillen, Laetitia; Savoye, Anne; Marchand, Virginie; Dubois, Agnès; Sanglier-Cianférani, Sarah; Van Dorsselaer, Alain; Clerc, Philippe; Avner, Philip; Visvikis, Athanase; Branlant, Christiane

    2010-01-01

    In placental mammals, inactivation of one of the X chromosomes in female cells ensures sex chromosome dosage compensation. The 17 kb non-coding Xist RNA is crucial to this process and accumulates on the future inactive X chromosome. The most conserved Xist RNA region, the A region, contains eight or nine repeats separated by U-rich spacers. It is implicated in the recruitment of late inactivated X genes to the silencing compartment and likely in the recruitment of complex PRC2. Little is known about the structure of the A region and more generally about Xist RNA structure. Knowledge of its structure is restricted to an NMR study of a single A repeat element. Our study is the first experimental analysis of the structure of the entire A region in solution. By the use of chemical and enzymatic probes and FRET experiments, using oligonucleotides carrying fluorescent dyes, we resolved problems linked to sequence redundancies and established a 2-D structure for the A region that contains two long stem-loop structures each including four repeats. Interactions formed between repeats and between repeats and spacers stabilize these structures. Conservation of the spacer terminal sequences allows formation of such structures in all sequenced Xist RNAs. By combination of RNP affinity chromatography, immunoprecipitation assays, mass spectrometry, and Western blot analysis, we demonstrate that the A region can associate with components of the PRC2 complex in mouse ES cell nuclear extracts. Whilst a single four-repeat motif is able to associate with components of this complex, recruitment of Suz12 is clearly more efficient when the entire A region is present. Our data with their emphasis on the importance of inter-repeat pairing change fundamentally our conception of the 2-D structure of the A region of Xist RNA and support its possible implication in recruitment of the PRC2 complex. PMID:20052282

  15. Development and Implementation of a Scramjet Cycle Analysis Code with a Finite-Rate-Chemistry Combustion Model for Use on a Personal Computer

    DTIC Science & Technology

    1993-12-01

    2 3 9 V List of Fi-ures Figure 1 - Functional...Block Diagram of a Scramjet ........................................ 9 Figure 2 - ’Corrected’ Specific Impulse of Hydrogen-Oxygen Rocket ............. 35...38 Figure 8 - Schematic of Northam/Anderson Mixing Model ............................ 39 Figure 9 - Pressure-Area

  16. Prevalence and associated risk factors of undercorrected refractive errors among people with diabetes in Shanghai.

    PubMed

    Zhu, Mengjun; Tong, Xiaowei; Zhao, Rong; He, Xiangui; Zhao, Huijuan; Zhu, Jianfeng

    2017-11-28

    To investigate the prevalence and risk factors of undercorrected refractive error (URE) among people with diabetes in the Baoshan District of Shanghai, where data for undercorrected refractive error are limited. The study was a population-based survey of 649 persons (aged 60 years or older) with diabetes in Baoshan, Shanghai in 2009. One copy of the questionnaire was completed for each subject. Examinations included a standardized refraction and measurement of presenting and best-corrected visual acuity (BCVA), tonometry, slit lamp biomicroscopy, and fundus photography. The calculated age-standardized prevalence rate of URE was 16.63% (95% confidence interval [CI] 13.76-19.49). For visual impairment subjects (presenting vision worse than 20/40 in the better eye), the prevalence of URE was up to 61.11%, and 75.93% of subjects could achieve visual acuity improvement by at least one line using appropriate spectacles. Under multiple logistic regression analysis, older age, female gender, non-farmer, increasing degree of myopia, lens opacities status, diabetic retinopathy (DR), body mass index (BMI) index lower than normal, and poor glycaemic control were associated with higher URE levels. Wearing distance eyeglasses was a protective factor for URE. The undercorrected refractive error in diabetic adults was high in Shanghai. Health education and regular refractive assessment are needed for diabetic adults. Persons with diabetes should be more aware that poor vision is often correctable, especially for those with risk factors.

  17. 50 CFR 2.2 - Locations of regional offices.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... the States of Arizona, New Mexico, Oklahoma, and Texas), 500 Gold Avenue SW., Room 9018 (P.O. Box 1306... States of Colorado, Kansas, Montana, Nebraska, North Dakota, South Dakota, Utah and Wyoming), 134 Union Boulevard (P.O. Box 25486), Lakewood, Colorado 80228. (g) Alaska Regional Office (Region 7—comprising the...

  18. Study on embodied CO2 transfer between the Jing-Jin-Ji region and other regions in China: a quantification using an interregional input-output model.

    PubMed

    Chen, Mengmeng; Wu, Sanmang; Lei, Yalin; Li, Shantong

    2018-05-01

    Jing-Jin-Ji region (i.e., Beijing, Tianjin, and Hebei) is China's key development region, but it is also the leading and most serious air pollution region in China. High fossil fuel consumption is the major source of both carbon dioxide (CO 2 ) emissions and air pollutants. Therefore, it is important to reveal the source of CO 2 emissions to control the air pollution in the Jing-Jin-Ji region. In this study, an interregional input-output model was applied to quantitatively estimate the embodied CO 2 transfer between Jing-Jin-Ji region and other region in China using China's interregional input-output data in 2010. The results indicated that there was a significant difference in the production-based CO 2 emissions in China, and furthermore, the Jing-Jin-Ji region and its surrounding regions were the main regions of the production-based CO 2 emissions in China. Hebei Province exported a large amount of embodied CO 2 to meet the investment, consumption, and export demands of Beijing and Tianjin. The Jing-Jin-Ji regions exported a great deal of embodied CO 2 to the coastal provinces of southeast China and imported it from neighboring provinces.

  19. Improved simulation of regional CO2 surface concentrations using GEOS-Chem and fluxes from VEGAS

    NASA Astrophysics Data System (ADS)

    Chen, Z. H.; Zhu, J.; Zeng, N.

    2013-08-01

    CO2 measurements have been combined with simulated CO2 distributions from a transport model in order to produce the optimal estimates of CO2 surface fluxes in inverse modeling. However, one persistent problem in using model-observation comparisons for this goal relates to the issue of compatibility. Observations at a single station reflect all underlying processes of various scales. These processes usually cannot be fully resolved by model simulations at the grid points nearest the station due to lack of spatial or temporal resolution or missing processes in the model. In this study the stations in one region were grouped based on the amplitude and phase of the seasonal cycle at each station. The regionally averaged CO2 at all stations in one region represents the regional CO2 concentration of this region. The regional CO2 concentrations from model simulations and observations were used to evaluate the regional model results. The difference of the regional CO2 concentration between observation and modeled results reflects the uncertainty of the large-scale flux in the region where the grouped stations are. We compared the regional CO2 concentrations between model results with biospheric fluxes from the Carnegie-Ames-Stanford Approach (CASA) and VEgetation-Global-Atmosphere-Soil (VEGAS) models, and used observations from GLOBALVIEW-CO2 to evaluate the regional model results. The results show the largest difference of the regionally averaged values between simulations with fluxes from VEGAS and observations is less than 5 ppm for North American boreal, North American temperate, Eurasian boreal, Eurasian temperate and Europe, which is smaller than the largest difference between CASA simulations and observations (more than 5 ppm). There is still a large difference between two model results and observations for the regional CO2 concentration in the North Atlantic, Indian Ocean, and South Pacific tropics. The regionally averaged CO2 concentrations will be helpful for

  20. 50 CFR 2.2 - Locations of regional offices.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ...—comprising the States of Arizona, New Mexico, Oklahoma and Texas), 500 Gold Avenue, SW, Room 9018 (P.O. Box... Regional Office (Region 6—comprising the States of Colorado, Kansas, Montana, Nebraska, North Dakota, South Dakota, Utah and Wyoming), 134 Union Boulevard (P.O. Box 25486), Denver Federal Center, Denver, Colorado...

  1. 50 CFR 2.2 - Locations of regional offices.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ...—comprising the States of Arizona, New Mexico, Oklahoma and Texas), 500 Gold Avenue, SW, Room 9018 (P.O. Box... Regional Office (Region 6—comprising the States of Colorado, Kansas, Montana, Nebraska, North Dakota, South Dakota, Utah and Wyoming), 134 Union Boulevard (P.O. Box 25486), Denver Federal Center, Denver, Colorado...

  2. 50 CFR 2.2 - Locations of regional offices.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ...—comprising the States of Arizona, New Mexico, Oklahoma and Texas), 500 Gold Avenue, SW, Room 9018 (P.O. Box... Regional Office (Region 6—comprising the States of Colorado, Kansas, Montana, Nebraska, North Dakota, South Dakota, Utah and Wyoming), 134 Union Boulevard (P.O. Box 25486), Denver Federal Center, Denver, Colorado...

  3. The UKC2 regional coupled environmental prediction system

    NASA Astrophysics Data System (ADS)

    Lewis, Huw W.; Castillo Sanchez, Juan Manuel; Graham, Jennifer; Saulter, Andrew; Bornemann, Jorge; Arnold, Alex; Fallmann, Joachim; Harris, Chris; Pearson, David; Ramsdale, Steven; Martínez-de la Torre, Alberto; Bricheno, Lucy; Blyth, Eleanor; Bell, Victoria A.; Davies, Helen; Marthews, Toby R.; O'Neill, Clare; Rumbold, Heather; O'Dea, Enda; Brereton, Ashley; Guihou, Karen; Hines, Adrian; Butenschon, Momme; Dadson, Simon J.; Palmer, Tamzin; Holt, Jason; Reynard, Nick; Best, Martin; Edwards, John; Siddorn, John

    2018-01-01

    It is hypothesized that more accurate prediction and warning of natural hazards, such as of the impacts of severe weather mediated through various components of the environment, require a more integrated Earth System approach to forecasting. This hypothesis can be explored using regional coupled prediction systems, in which the known interactions and feedbacks between different physical and biogeochemical components of the environment across sky, sea and land can be simulated. Such systems are becoming increasingly common research tools. This paper describes the development of the UKC2 regional coupled research system, which has been delivered under the UK Environmental Prediction Prototype project. This provides the first implementation of an atmosphere-land-ocean-wave modelling system focussed on the United Kingdom and surrounding seas at km-scale resolution. The UKC2 coupled system incorporates models of the atmosphere (Met Office Unified Model), land surface with river routing (JULES), shelf-sea ocean (NEMO) and ocean waves (WAVEWATCH III). These components are coupled, via OASIS3-MCT libraries, at unprecedentedly high resolution across the UK within a north-western European regional domain. A research framework has been established to explore the representation of feedback processes in coupled and uncoupled modes, providing a new research tool for UK environmental science. This paper documents the technical design and implementation of UKC2, along with the associated evaluation framework. An analysis of new results comparing the output of the coupled UKC2 system with relevant forced control simulations for six contrasting case studies of 5-day duration is presented. Results demonstrate that performance can be achieved with the UKC2 system that is at least comparable to its component control simulations. For some cases, improvements in air temperature, sea surface temperature, wind speed, significant wave height and mean wave period highlight the potential

  4. Water masers and ammonia (1, 1) and (2, 2) towards six regions in the Carina Nebula

    NASA Astrophysics Data System (ADS)

    Breen, S. L.; Green, C.-E.; Cunningham, M. R.; Voronkov, M. A.; Horiuchi, S.; Green, J. A.

    2018-01-01

    We present water maser and ammonia (1, 1) and (2, 2) observations, towards six regions in the Carina Nebula, conducted with the Australia Telescope Compact Array. In total five water masers were detected within two of the target fields, and we provide their accurate positions and characteristics. These five water masers constitute all of the known masers detected towards star formation regions in the Carina Nebula and we argue, that given the evidence for active star formation, and the presence of many high-mass stars, the Carina Nebula is uncharacteristically devoid of masers. Our results are consistent with the Carina Nebula having a lack of young high-mass stars, despite the presence of older high-mass stars. Ammonia (1, 1) and (2, 2) emission was detected towards all but one of the target fields and we find that their linewidths and derived temperatures are consistent with the presence of young star formation regions.

  5. Ammon's Horn 2 (CA2) of the Hippocampus: A Long-Known Region with a New Potential Role in Neurodegeneration.

    PubMed

    Pang, Cindy Chi-Ching; Kiecker, Clemens; O'Brien, John T; Noble, Wendy; Chang, Raymond Chuen-Chung

    2018-06-01

    The hippocampus has a critical role in cognition and human memory and is one of the most studied structures in the brain. Despite more than 400 years of research, little is known about the Ammon's horn region cornu ammonis 2 (CA2) subfield in comparison to other subfield regions (CA1, CA3, and CA4). Recent findings have shown that CA2 plays a bigger role than previously thought. Here, we review understanding of hippocampus and CA2 ontogenesis, together with basic and clinical findings about the potential role of this region in neurodegenerative disease. The CA2 has widespread anatomical connectivity, unique signaling molecules, and intrinsic electrophysiological properties. Experimental studies using in vivo models found that the CA2 region has a role in cognition, especially in social memory and object recognition. In models of epilepsy and hypoxia, the CA2 exhibits higher resilience to cell death and hypoxia in comparison with neighboring regions, and while hippocampal atrophy remains poorly understood in Parkinson's disease (PD) and dementia with Lewy bodies (DLB), findings from postmortem PD brain demonstrates clear accumulation of α-synuclein pathology in CA2, and the CA2-CA3 region shows relatively more atrophy compared with other hippocampal subfields. Taken together, there is a growing body of evidence suggesting that the CA2 can be an ideal hallmark with which to differentiate different neurodegenerative stages of PD. Here, we summarize these recent data and provide new perspectives/ideas for future investigations to unravel the contribution of the CA2 to neurodegenerative diseases.

  6. 78 FR 71448 - Regional Reliability Standard BAL-002-WECC-2-Contingency Reserve

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-11-29

    ... DEPARTMENT OF ENERGY Federal Energy Regulatory Commission 18 CFR Part 40 [Docket No. RM13-13-000; Order No. 789] Regional Reliability Standard BAL-002-WECC-2--Contingency Reserve AGENCY: Federal Energy..., and Tony Clark. Regional Reliability Standard BAL-002-WECC-2--Contingency Reserve Docket No. RM13-13...

  7. Cholecystokinin levels in prohormone convertase 2 knock-out mouse brain regions reveal a complex phenotype of region-specific alterations.

    PubMed

    Beinfeld, Margery C; Blum, Alissa; Vishnuvardhan, Daesety; Fanous, Sanya; Marchand, James E

    2005-11-18

    Prohormone convertase 2 is widely co-localized with cholecystokinin in rodent brain. To examine its role in cholecystokinin processing, cholecystokinin levels were measured in dissected brain regions from prohormone convertase 2 knock-out mice. Cholecystokinin levels were lower in hippocampus, septum, thalamus, mesencephalon, and pons in knock-out mice than wild-type mice. In cerebral cortex, cortex-related structures and olfactory bulb, cholecystokinin levels were higher than wild type. Female mice were more affected by the loss of prohormone convertase 2 than male mice. The decrease in cholecystokinin levels in these brain regions shows that prohormone convertase 2 is important for cholecystokinin processing. Quantitative polymerase chain reaction measurements were performed to examine the relationship between peptide levels and cholecystokinin and enzyme expression. They revealed that cholecystokinin and prohormone convertase 1 mRNA levels in cerebral cortex and olfactory bulb were actually lower in knock-out than wild type, whereas their expression in other brain regions of knock-out mouse brain was the same as wild type. Female mice frequently had higher expression of cholecystokinin and prohormone convertase 1, 2, and 5 mRNA than male mice. The loss of prohormone convertase 2 alters CCK processing in specific brain regions. This loss also appears to trigger compensatory mechanisms in cerebral cortex and olfactory bulb that produce elevated levels of cholecystokinin but do not involve increased expression of cholecystokinin, prohormone convertase 1 or 5 mRNA.

  8. Potential for 4-n-nonylphenol biodegradation in stream sediments

    USGS Publications Warehouse

    Bradley, P.M.; Barber, L.B.; Kolpin, D.W.; McMahon, P.B.; Chapelle, F.H.

    2008-01-01

    The potential for in situ biodegradation of 4-nonylphenol (4-NP) was investigated in three hydrologically distinct streams impacted by wastewater treatment plants (WWTPs) in the United States. Microcosms were prepared with sediments from each site and amended with [U-ring-14C]4-n-nonylphenol (4-n-NP) as a model test substrate. Microcosms prepared with sediment collected upstream of the WWTP outfalls and incubated under oxic conditions showed rapid and complete mineralization of [U-ring-14C]4- n-NP to 14CO2 in all three systems. In contrast, no mineralization of [U-ring-14C]4-n-NP was observed in these sediments under anoxic (methanogenic) conditions. The initial linear rate of [U-ring-14C]4-n-NP mineralization in sediments from upstream and downstream of the respective WWTP outfalls was inversely correlated with the biochemical oxygen demand (BOD) of the streambed sediments. These results suggest that the net supply of dissolved oxygen to streambed sediments is a key determinant of the rate and extent of 4-NP biodegradation in stream systems. In the stream systems considered by the present study, dissolved oxygen concentrations in the overlying water column (8–10 mg/L) and in the bed sediment pore water (1–3 mg/L at a depth of 10 cm below the sediment–water interface) were consistent with active in situ 4-NP biodegradation. These results suggest WWTP procedures that maximize the delivery of dissolved oxygen while minimizing the release of BOD to stream receptors favor efficient biodegradation of 4-NP contaminants in wastewater-impacted stream environments.

  9. Recruitment and Recommendation of College Students: North Korea.

    DTIC Science & Technology

    1960-09-14

    Journalism, Chinese Literat- ure, Library Science , Russian Language and Literature, Geology, Geodesic Chart Science (Ch’ukchi Chldohak), Planning...History, Philosophy, Library Science , Politi- cal Economy, and Meteorological ^um.unhak/. (f) The Preparatory School for Honor Soldiers. 2. Kim

  10. TMI-2 (Three Mile Island Unit 2) core region defueling

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Rodabaugh, J.M.; Cowser, D.K.

    1988-01-01

    In July of 1982, a video camera was inserted into the Three Mile Island Unit 2 reactor vessel providing the first visual evidence of core damage. This inspection, and numerous subsequent data acquisition tasks, revealed a central void /approx/1.5 m (5 ft) deep. This void region was surrounded by partial length fuel assemblies and ringed on the periphery by /approx/40 full-length, but partial cross-section, fuel assemblies. All of the original 177 fuel assemblies exhibited signs of damage. The bottom of the void cavity was covered with a bed of granular rubble, fuel assembly upper end fittings, control rod spiders, fuelmore » rod fragments, and fuel pellets. It was obvious that the normal plant refueling system not suitable for removing the damaged core. A new system of defueling tools and equipment was necessary to perform this task. Design of the new system was started immediately, followed by >1 yr of fabrication. Delivery and checkout of the defueling system occurred in mid-1985. Actual defueling was initiated in late 1985 with removal of the debris bed at the bottom of the core void. Obstructions to the debris, such as end fittings and fuel rod fragments ere removed first; then /approx/23,000 kg (50,000lb) of granular debris was quickly loaded into canisters. Core region defueling was completed in late 1987, /approx/2 yr after it was initiated.« less

  11. Using Climate Regionalization to Understand Climate Forecast System Version 2 (CFSv2) Precipitation Performance for the Conterminous United States (CONUS)

    NASA Technical Reports Server (NTRS)

    Regonda, Satish K.; Zaitchik, Benjamin F.; Badr, Hamada S.; Rodell, Matthew

    2016-01-01

    Dynamically based seasonal forecasts are prone to systematic spatial biases due to imperfections in the underlying global climate model (GCM). This can result in low-forecast skill when the GCM misplaces teleconnections or fails to resolve geographic barriers, even if the prediction of large-scale dynamics is accurate. To characterize and address this issue, this study applies objective climate regionalization to identify discrepancies between the Climate Forecast SystemVersion 2 (CFSv2) and precipitation observations across the Contiguous United States (CONUS). Regionalization shows that CFSv2 1 month forecasts capture the general spatial character of warm season precipitation variability but that forecast regions systematically differ from observation in some transition zones. CFSv2 predictive skill for these misclassified areas is systematically reduced relative to correctly regionalized areas and CONUS as a whole. In these incorrectly regionalized areas, higher skill can be obtained by using a regional-scale forecast in place of the local grid cell prediction.

  12. Planning of the 12 GHz broadcasting satellite service in Region 2

    NASA Astrophysics Data System (ADS)

    Bowen, R. R.; Chambers, J. G.

    The 12 GHz band broadcasting satellite plan formulated for Region 2 (the Americas) at the 1983 Regional Administrative Radio Conference met all requirements with a total of 1144 assigned channels. In order to achieve this high capacity (16 channels/service area), it was necessary to place some assignments at orbit locations with relatively low elevation angles and early eclipse times. This high capacity does not prevent the plan from adapting to changing technological features of broadcasting in this band as it evolves over the years, and is estimated to be more than 2.5 times larger than the capacity which would be available without taking advantage of the geographical characteristics of the region, as well as of isolation by means of spacecraft antennas.

  13. Vacuum ultraviolet photoabsorption spectroscopy of CH2Cl2 and CD2Cl2 in the energy region 50,000-95,000 cm-1

    NASA Astrophysics Data System (ADS)

    Mandal, Anuvab; Singh, Param Jeet; Shastri, Aparna; Jagatap, B. N.

    2014-12-01

    A consolidated study of the VUV absorption spectra of CH2Cl2 and CD2Cl2 in the 50,000-95,000 cm-1 region using synchrotron radiation is presented. Rydberg series and vibronic analysis are carried out and supported by quantum chemical calculations. The broad absorption band of CH2Cl2 in the region 50,000-60,000 cm-1 is attributed to the valence states 11B2, 11B1 and 11A1. Most of the bands in the 60,000-95,000 cm-1 region are fitted to Rydberg series of ns, np and nd type converging to the first four ionization potentials 11.320, 11.357, 12.152 and 12.271 eV of CH2Cl2 arising from excitation of an electron from one of the four outermost Cl non-bonding orbitals (2b1, 3b2, 1a2 and 4a1). Vertical excited states of CH2Cl2 calculated using TDDFT are correlated with experimentally observed electronic states based on the symmetries of the initial and final MOs involved in a transition. A few Rydberg transitions viz. 2b1→5s, 4p, 5p, 6p; 3b2→4p, 5p; 1a2→4p are accompanied by vibronic features. Observed vibronic bands are assigned mainly to the CCl symmetric stretch (ν3‧) mode with smaller contributions from the CH symmetric stretch (ν1‧), CH2 bend (ν2‧) and CH2 wag (ν8‧) modes. Assignments are corroborated by comparison with the VUV absorption spectrum of the deuterated isotopologue CD2Cl2, reported here for the first time. The high underlying intensities seen in several sub-regions are explained by valence or valence-Rydberg mixed type transitions predicted with high oscillator strengths by the TDDFT calculations.

  14. Improved influenza viral vector based Brucella abortus vaccine induces robust B and T-cell responses and protection against Brucella melitensis infection in pregnant sheep and goats

    PubMed Central

    Mailybayeva, Aigerim; Yespembetov, Bolat; Ryskeldinova, Sholpan; Zinina, Nadezhda; Sansyzbay, Abylai; Renukaradhya, Gourapura J.; Petrovsky, Nikolai

    2017-01-01

    We previously developed a potent candidate vaccine against bovine brucellosis caused by Brucella abortus using the influenza viral vector expressing Brucella Omp16 and L7/L12 proteins (Flu-BA). Our success in the Flu-BA vaccine trial in cattle and results of a pilot study in non-pregnant small ruminants prompted us in the current study to test its efficacy against B. melitensis infection in pregnant sheep and goats. In this study, we improved the Flu-BA vaccine formulation and immunization method to achieve maximum efficacy and safety. The Flu-BA vaccine formulation had two additional proteins Omp19 and SOD, and administered thrice with 20% Montanide Gel01 adjuvant, simultaneously by both subcutaneous and conjunctival routes at 21 days intervals in pregnant sheep and goats. At 42 days post-vaccination (DPV) we detected antigen-specific IgG antibodies predominantly of IgG2a isotype but also IgG1, and also detected a strong lymphocyte recall response with IFN-γ production. Importantly, our candidate vaccine prevented abortion in 66.7% and 77.8% of pregnant sheep and goats, respectively. Furthermore, complete protection (absence of live B. melitensis 16M) was observed in 55.6% and 66.7% of challenged sheep and goats, and 72.7% and 90.0% of their fetuses (lambs/yeanlings), respectively. The severity of B. melitensis 16M infection in vaccinated sheep and goats and their fetuses (index of infection and rates of Brucella colonization in tissues) was significantly lower than in control groups. None of the protection parameters after vaccination with Flu-BA vaccine were statistically inferior to protection seen with the commercial B. melitensis Rev.1 vaccine (protection against abortion and vaccination efficacy, alpha = 0.18–0.34, infection index, P = 0.37–0.77, Brucella colonization, P = 0.16 to P > 0.99). In conclusion, our improved Flu-BA vaccine formulation and delivery method were found safe and effective in protecting pregnant sheep and goats against adverse

  15. Improved influenza viral vector based Brucella abortus vaccine induces robust B and T-cell responses and protection against Brucella melitensis infection in pregnant sheep and goats.

    PubMed

    Mailybayeva, Aigerim; Yespembetov, Bolat; Ryskeldinova, Sholpan; Zinina, Nadezhda; Sansyzbay, Abylai; Renukaradhya, Gourapura J; Petrovsky, Nikolai; Tabynov, Kaissar

    2017-01-01

    We previously developed a potent candidate vaccine against bovine brucellosis caused by Brucella abortus using the influenza viral vector expressing Brucella Omp16 and L7/L12 proteins (Flu-BA). Our success in the Flu-BA vaccine trial in cattle and results of a pilot study in non-pregnant small ruminants prompted us in the current study to test its efficacy against B. melitensis infection in pregnant sheep and goats. In this study, we improved the Flu-BA vaccine formulation and immunization method to achieve maximum efficacy and safety. The Flu-BA vaccine formulation had two additional proteins Omp19 and SOD, and administered thrice with 20% Montanide Gel01 adjuvant, simultaneously by both subcutaneous and conjunctival routes at 21 days intervals in pregnant sheep and goats. At 42 days post-vaccination (DPV) we detected antigen-specific IgG antibodies predominantly of IgG2a isotype but also IgG1, and also detected a strong lymphocyte recall response with IFN-γ production. Importantly, our candidate vaccine prevented abortion in 66.7% and 77.8% of pregnant sheep and goats, respectively. Furthermore, complete protection (absence of live B. melitensis 16M) was observed in 55.6% and 66.7% of challenged sheep and goats, and 72.7% and 90.0% of their fetuses (lambs/yeanlings), respectively. The severity of B. melitensis 16M infection in vaccinated sheep and goats and their fetuses (index of infection and rates of Brucella colonization in tissues) was significantly lower than in control groups. None of the protection parameters after vaccination with Flu-BA vaccine were statistically inferior to protection seen with the commercial B. melitensis Rev.1 vaccine (protection against abortion and vaccination efficacy, alpha = 0.18-0.34, infection index, P = 0.37-0.77, Brucella colonization, P = 0.16 to P > 0.99). In conclusion, our improved Flu-BA vaccine formulation and delivery method were found safe and effective in protecting pregnant sheep and goats against adverse

  16. Global and regional emissions estimates for N2O

    NASA Astrophysics Data System (ADS)

    Saikawa, E.; Prinn, R. G.; Dlugokencky, E.; Ishijima, K.; Dutton, G. S.; Hall, B. D.; Langenfelds, R.; Tohjima, Y.; Machida, T.; Manizza, M.; Rigby, M.; O'Doherty, S.; Patra, P. K.; Harth, C. M.; Weiss, R. F.; Krummel, P. B.; van der Schoot, M.; Fraser, P. J.; Steele, L. P.; Aoki, S.; Nakazawa, T.; Elkins, J. W.

    2014-05-01

    We present a comprehensive estimate of nitrous oxide (N2O) emissions using observations and models from 1995 to 2008. High-frequency records of tropospheric N2O are available from measurements at Cape Grim, Tasmania; Cape Matatula, American Samoa; Ragged Point, Barbados; Mace Head, Ireland; and at Trinidad Head, California using the Advanced Global Atmospheric Gases Experiment (AGAGE) instrumentation and calibrations. The Global Monitoring Division of the National Oceanic and Atmospheric Administration/Earth System Research Laboratory (NOAA/ESRL) has also collected discrete air samples in flasks and in situ measurements from remote sites across the globe and analyzed them for a suite of species including N2O. In addition to these major networks, we include in situ and aircraft measurements from the National Institute of Environmental Studies (NIES) and flask measurements from the Tohoku University and Commonwealth Scientific and Industrial Research Organization (CSIRO) networks. All measurements show increasing atmospheric mole fractions of N2O, with a varying growth rate of 0.1-0.7% per year, resulting in a 7.4% increase in the background atmospheric mole fraction between 1979 and 2011. Using existing emission inventories as well as bottom-up process modeling results, we first create globally gridded a priori N2O emissions over the 37 years since 1975. We then use the three-dimensional chemical transport model, Model for Ozone and Related Chemical Tracers version 4 (MOZART v4), and a Bayesian inverse method to estimate global as well as regional annual emissions for five source sectors from 13 regions in the world. This is the first time that all of these measurements from multiple networks have been combined to determine emissions. Our inversion indicates that global and regional N2O emissions have an increasing trend between 1995 and 2008. Despite large uncertainties, a significant increase is seen from the Asian agricultural sector in recent years, most likely

  17. Comparison of Radio Frequency Distinct Native Attribute and Matched Filtering Techniques for Device Discrimination and Operation Identification

    DTIC Science & Technology

    identification. URE from ten MSP430F5529 16-bit microcontrollers were analyzed using: 1) RF distinct native attributes (RF-DNA) fingerprints paired with multiple...discriminant analysis/maximum likelihood (MDA/ML) classification, 2) RF-DNA fingerprints paired with generalized relevance learning vector quantized

  18. Cell biology of Smad2/3 linker region phosphorylation in vascular smooth muscle.

    PubMed

    Rezaei, Hossein B; Kamato, Danielle; Ansari, Ghazaleh; Osman, Narin; Little, Peter J

    2012-08-01

    The transforming growth factor (TGF)-β superfamily of ligands regulates a diverse set of cellular functions. Transforming growth factor-β induces its biological effects through Type I and Type II transmembrane receptors that have serine/threonine kinase activities and weak tyrosine kinase activity. In vascular smooth muscle, TGF-β binds to the TGF-β Type II receptor (TβRII) at the cell surface, recruiting the Type I receptor (TβRI) to form a heterocomplex. Consequently, after phosphorylation and activation of TβRI, the transcription factors receptor activated (R-) Smad2 and Smad3 are recruited and activated through phosphorylation of C terminal residues. Overall, Smad2/3 and co-Smad4 have similar structures consisting of three regions an N-terminal MH1 domain, a C-terminal MH2 domain and a central linker region. Phosphorylation of the Smad linker region appears to have an important role in the regulation of Smad activity and function. The mitogen-activated protein kinase (MAPK) family, CDK2, CDK4 and calcium-calmodulin dependent kinase are the main kinases that phosphorylate sites in the linker region. The role of the linker region includes enabling the formation of Smad homo-oligomers and provision of phosphorylation sites for MAPK and other kinases. In some instances, linker region phosphorylation regulates the inhibition of the nuclear translocation of Smads. In the present review, we describe TGF-β signalling through Smad2/3 and the importance of the linker region in the regulation and expression of genes induced by TGF-β superfamily ligands in the context of vascular smooth muscle. © 2011 The Authors. Clinical and Experimental Pharmacology and Physiology © 2011 Blackwell Publishing Asia Pty Ltd.

  19. High Velocity Jet Noise Source Location and Reduction. Task 2. Theoretical Developments and Basic Experiments.

    DTIC Science & Technology

    1978-05-01

    Measured Flight Effect for J85/ Aerotrain Conical Nozzle, 400 ft Sideline. 360 4-149. Comparison of Predicted and Measured Flight Velocity Exponent m for J85... Aerotrain Conical Nozzle. 362 4-150. Comparison of Measured and Predicted Flight Noise Spectra for J85/ Aerotrain Conical Nozzle, V = 2200 fps, 400 ft...Bertin Aerotrain simulated flight noise results which were obtained by Clapper, et al.( 72) in Task 4 of this program. Fig- ure 4-148 shows the

  20. The role of ExbD in periplasmic pH homeostasis in Helicobacter pylori

    PubMed Central

    Marcus, Elizabeth A.; Sachs, George; Scott, David R.

    2013-01-01

    Background Helicobacter pylori, a neutralophile, colonizes the acidic environment of the human stomach by employing acid acclimation mechanisms that regulate periplasmic and cytoplasmic pH. The regulation of urease activity is central to acid acclimation. Inactive urease apoenzyme, UreA/B, requires nickel for activation. Accessory proteins UreE, F, G and H are required for nickel insertion into apoenzyme. The ExbB/ExbD/TonB complex transfers energy from the inner to outer membrane, providing the driving force for nickel uptake. Therefore, the aim of this study was to determine the contribution of ExbD to pH homeostasis. Materials and Methods A nonpolar exbD knockout was constructed and survival, growth, urease activity, and membrane potential were determined in comparison to wildtype. Results Survival of the ΔexbD strain was significantly reduced at pH 3.0. Urease activity as a function of pH and UreI activation were similar to the wildtype strain, showing normal function of the proton-gated urea channel, UreI. The increase in total urease activity over time in acid seen in the wildtype strain was abolished in the ΔexbD strain, but recovered in the presence of supra-physiologic nickel concentrations, demonstrating that the effect of the ΔexbD mutant is due to loss of a necessary constant supply of nickel. In acid, ΔexbD also decreased its ability to maintain membrane potential and periplasmic buffering in the presence of urea. Conclusions ExbD is essential for maintenance of periplasmic buffering and membrane potential by transferring energy required for nickel uptake, making it a potential non-antibiotic target for H. pylori eradication. PMID:23600974

  1. An approach to optimize the batch mixing process for improving the quality consistency of the products made from traditional Chinese medicines*

    PubMed Central

    Yan, Bin-jun; Qu, Hai-bin

    2013-01-01

    The efficacy of traditional Chinese medicine (TCM) is based on the combined effects of its constituents. Variation in chemical composition between batches of TCM has always been the deterring factor in achieving consistency in efficacy. The batch mixing process can significantly reduce the batch-to-batch quality variation in TCM extracts by mixing them in a well-designed proportion. However, reducing the quality variation without sacrificing too much of the production efficiency is one of the challenges. Accordingly, an innovative and practical batch mixing method aimed at providing acceptable efficiency for industrial production of TCM products is proposed in this work, which uses a minimum number of batches of extracts to meet the content limits. The important factors affecting the utilization ratio of the extracts (URE) were studied by simulations. The results have shown that URE was affected by the correlation between the contents of constituents, and URE decreased with the increase in the number of targets and the relative standard deviations of the contents. URE could be increased by increasing the number of storage tanks. The results have provided a reference for designing the batch mixing process. The proposed method has possible application value in reducing the quality variation in TCM and providing acceptable production efficiency simultaneously. PMID:24190450

  2. An approach to optimize the batch mixing process for improving the quality consistency of the products made from traditional Chinese medicines.

    PubMed

    Yan, Bin-jun; Qu, Hai-bin

    2013-11-01

    The efficacy of traditional Chinese medicine (TCM) is based on the combined effects of its constituents. Variation in chemical composition between batches of TCM has always been the deterring factor in achieving consistency in efficacy. The batch mixing process can significantly reduce the batch-to-batch quality variation in TCM extracts by mixing them in a well-designed proportion. However, reducing the quality variation without sacrificing too much of the production efficiency is one of the challenges. Accordingly, an innovative and practical batch mixing method aimed at providing acceptable efficiency for industrial production of TCM products is proposed in this work, which uses a minimum number of batches of extracts to meet the content limits. The important factors affecting the utilization ratio of the extracts (URE) were studied by simulations. The results have shown that URE was affected by the correlation between the contents of constituents, and URE decreased with the increase in the number of targets and the relative standard deviations of the contents. URE could be increased by increasing the number of storage tanks. The results have provided a reference for designing the batch mixing process. The proposed method has possible application value in reducing the quality variation in TCM and providing acceptable production efficiency simultaneously.

  3. Level IV Ecoregions of EPA Region 2

    EPA Pesticide Factsheets

    Ecoregions by EPA region were extracted from the seamless national shapefile. Ecoregions denote areas of general similarity in ecosystems and in the type, quality, and quantity of environmental resources. They are designed to serve as a spatial framework for the research, assessment, management, and monitoring of ecosystems and ecosystem components. These general purpose regions are critical for structuring and implementing ecosystem management strategies across federal agencies, state agencies, and nongovernment organizations that are responsible for different types of resources within the same geographical areas. The approach used to compile this map is based on the premise that ecological regions can be identified through the analysis of patterns of biotic and abiotic phenomena, including geology, physiography, vegetation, climate, soils, land use, wildlife, and hydrology. The relative importance of each characteristic varies from one ecological region to another. A Roman numeral hierarchical scheme has been adopted for different levels for ecological regions. Level I is the coarsest level, dividing North America into 15 ecological regions. Level II divides the continent into 52 regions (Commission for Environmental Cooperation Working Group, 1997). At Level III, the continental United States contains 104 regions whereas the conterminous United States has 84 (U.S. Environmental Protection Agency, 2005). Level IV ecoregions are further subdivisions of Level I

  4. Level III Ecoregions of EPA Region 2

    EPA Pesticide Factsheets

    Ecoregions by EPA region were extracted from the seamless national shapefile. Ecoregions denote areas of general similarity in ecosystems and in the type, quality, and quantity of environmental resources. They are designed to serve as a spatial framework for the research, assessment, management, and monitoring of ecosystems and ecosystem components. These general purpose regions are critical for structuring and implementing ecosystem management strategies across federal agencies, state agencies, and nongovernment organizations that are responsible for different types of resources within the same geographical areas. The approach used to compile this map is based on the premise that ecological regions can be identified through the analysis of patterns of biotic and abiotic phenomena, including geology, physiography, vegetation, climate, soils, land use, wildlife, and hydrology. The relative importance of each characteristic varies from one ecological region to another. A Roman numeral hierarchical scheme has been adopted for different levels for ecological regions. Level I is the coarsest level, dividing North America into 15 ecological regions. Level II divides the continent into 52 regions (Commission for Environmental Cooperation Working Group, 1997). At Level III, the continental United States contains 104 regions whereas the conterminous United States has 84 (U.S. Environmental Protection Agency, 2005). Level IV ecoregions are further subdivisions of Level I

  5. Effect of Time on Perceived Gains from an Undergraduate Research Program

    PubMed Central

    Adedokun, Omolola A.; Parker, Loran C.; Childress, Amy; Burgess, Wilella; Adams, Robin; Agnew, Christopher R.; Leary, James; Knapp, Deborah; Shields, Cleveland; Lelievre, Sophie; Teegarden, Dorothy

    2014-01-01

    The current study examines the trajectories of student perceived gains as a result of time spent in an undergraduate research experience (URE). Data for the study come from a survey administered at three points over a 1-yr period: before participation in the program, at the end of a Summer segment of research, and at the end of the year. Repeated-measures analysis of variance was used to examine the effect of time on perceived gains in student research skills, research confidence, and understanding of research processes. The results suggest that the students experienced different gains/benefits at developmentally different stages of their UREs. Participants reported gains in fewer areas at the end of the Summer segment compared with the end of the yearlong experience, thus supporting the notion that longer UREs offer students more benefit. PMID:24591512

  6. Research notes : pavement markings using waterborne paint and Visibeads in Region 2.

    DOT National Transportation Integrated Search

    1995-01-01

    In order to address the problems associated with solvent-borne patins, ODOT Region 2 staff bought sufficient waterborne paint to place approximately 760 linear miles of stripes. This enabled the Region staff to evaluate state-of-the-art materials and...

  7. Distribution of Region 1 and 2 currents in the quietand substorm time plasma sheetfrom THEMIS observations

    NASA Astrophysics Data System (ADS)

    Liu, J.; Angelopoulos, V.; Chu, X.; McPherron, R. L.

    2016-12-01

    Although Earth's Region 1 and 2 currents are related to activities such as substorm initiation, their magnetospheric origin remains unclear. Utilizing the triangular configuration of THEMIS probes at 8-12 RE downtail, we seek the origin of nightside Region 1 and 2 currents. The triangular configuration allows a curlometer-like technique which do not rely on active-time boundary crossings, so we can examine the current distribution in quiet times as well as active times. Our statistical study reveals that both Region 1 and 2 currents exist in the plasma sheet during quiet and active times. Especially, this is the first unequivocal, in-situ evidence of the existence of Region 2 currents in the plasma sheet. Farther away from the neutral sheet than the Region 2 currents lie the Region 1 currents which extend at least to the plasma sheet boundary layer. At geomagnetic quiet times, the separation between the two currents is located 2.5 RE from the neutral sheet. These findings suggest that the plasma sheet is a source of Region 1 and 2 currents regardless of geomagnetic activity level. During substorms, the separation between Region 1 and 2 currents migrates toward (away from) the neutral sheet as the plasma sheet thins (thickens). This migration indicates that the deformation of Region 1 and 2 currents is associated with redistribution of FAC sources in the magnetotail. In some substorms when the THEMIS probes encounter a dipolarization, a substorm current wedge (SCW) can be inferred from our technique, and it shows a distinctively larger current density than the pre-existing Region 1 currents. This difference suggests that the SCW is not just an enhancement of the pre-existing Region 1 current; the SCW and the Region 1 currents have different sources.

  8. The Number, Organization, and Size of Polymorphic Membrane Protein Coding Sequences as well as the Most Conserved Pmp Protein Differ within and across Chlamydia Species.

    PubMed

    Van Lent, Sarah; Creasy, Heather Huot; Myers, Garry S A; Vanrompay, Daisy

    2016-01-01

    Variation is a central trait of the polymorphic membrane protein (Pmp) family. The number of pmp coding sequences differs between Chlamydia species, but it is unknown whether the number of pmp coding sequences is constant within a Chlamydia species. The level of conservation of the Pmp proteins has previously only been determined for Chlamydia trachomatis. As different Pmp proteins might be indispensible for the pathogenesis of different Chlamydia species, this study investigated the conservation of Pmp proteins both within and across C. trachomatis,C. pneumoniae,C. abortus, and C. psittaci. The pmp coding sequences were annotated in 16 C. trachomatis, 6 C. pneumoniae, 2 C. abortus, and 16 C. psittaci genomes. The number and organization of polymorphic membrane coding sequences differed within and across the analyzed Chlamydia species. The length of coding sequences of pmpA,pmpB, and pmpH was conserved among all analyzed genomes, while the length of pmpE/F and pmpG, and remarkably also of the subtype pmpD, differed among the analyzed genomes. PmpD, PmpA, PmpH, and PmpA were the most conserved Pmp in C. trachomatis,C. pneumoniae,C. abortus, and C. psittaci, respectively. PmpB was the most conserved Pmp across the 4 analyzed Chlamydia species. © 2016 S. Karger AG, Basel.

  9. Method for high-accuracy reflectance measurements in the 2.5-microm region.

    PubMed

    Richter, Rudolf; Müller, Andreas

    2003-02-20

    Reflectance measurement with spectroradiometers in the solar wavelength region (0.4-2.5 microm) are frequently conducted in the laboratory or in the field to characterize surface materials of artificial and natural targets. The spectral surface reflectance is calculated as the ratio of the signals obtained over the target surface and a reference panel, yielding a relative reflectance value. If the reflectance of the reference panel is known, the absolute target reflectance can be computed. This standard measurement technique assumes that the signal at the radiometer is due completely to reflected target and reference radiation. However, for field measurements in the 2.4-2.5-microm region with the Sun as the illumination source, the emitted thermal radiation is not a negligible part of the signal even at ambient temperatures, because the atmospheric transmittance, and thus the solar illumination level, is small in the atmospheric absorption regions. A new method is proposed that calculates reflectance values in the 2.4-2.5-microm region while it accounts for the reference panel reflectance and the emitted radiation. This technique needs instruments with noise-equivalent radiances of 2 orders of magnitude below currently commercially available instruments and requires measurement of the surface temperatures of target and reference. If the reference panel reflectance and temperature effects are neglected, the standard method yields reflectance errors up to 0.08 and 0.15 units for 7- and 2-nm bandwidth instruments, respectively. For the new method the corresponding errors can be reduced to approximately 0.01 units for the surface temperature range of 20-35 degrees C.

  10. Prevalence of Brucella abortus antibodies in equines of a tropical region of Mexico

    PubMed Central

    Acosta-González, Rosa I.; González-Reyes, Ismael; Flores-Gutiérrez, Gerardo H.

    2006-01-01

    A cross-sectional study was conducted to determinate the seroprevalence rate of equine brucellosis in the state of Tamaulipas, Mexico. Serum samples from 420 equines were analyzed with the Rose Bengal test at cell concentrations of 3% (RBT-3%) and 8% (RBT-8%), and positive results were confirmed with the Rivanol test (RT). Risk factors were determined with the prevalence ratio (PR) and the use of variables generated from a questionnaire administered to the animals’ owners. Serum from 1 stallion had positive results with both the RBT-8% and the RT, for a seroprevalence rate of 0.238%. Drinking of water from a pond that was also used by cattle and dogs was the only associated risk factor for this animal (PR = 0.25). However, the results were considered false-positive, because the results for other horses in the same environmental conditions were negative. Although brucellosis is considered endemic in ruminants in the study area, the results obtained suggest that equines are not a reservoir of brucellosis and do not play an important role in the epidemiologic patterns of this disease in northeastern Mexico. PMID:17042384

  11. Star Formation Activity in the Galactic H II Region Sh2-297

    NASA Astrophysics Data System (ADS)

    Mallick, K. K.; Ojha, D. K.; Samal, M. R.; Pandey, A. K.; Bhatt, B. C.; Ghosh, S. K.; Dewangan, L. K.; Tamura, M.

    2012-11-01

    We present a multiwavelength study of the Galactic H II region Sh2-297, located in the Canis Major OB1 complex. Optical spectroscopic observations are used to constrain the spectral type of ionizing star HD 53623 as B0V. The classical nature of this H II region is affirmed by the low values of electron density and emission measure, which are calculated to be 756 cm-3 and 9.15 × 105 cm-6 pc using the radio continuum observations at 610 and 1280 MHz, and Very Large Array archival data at 1420 MHz. To understand local star formation, we identified the young stellar object (YSO) candidates in a region of area ~7farcm5 × 7farcm5 centered on Sh2-297 using grism slitless spectroscopy (to identify the Hα emission line stars), and near infrared (NIR) observations. NIR YSO candidates are further classified into various evolutionary stages using color-color and color-magnitude (CM) diagrams, giving 50 red sources (H - K > 0.6) and 26 Class II-like sources. The mass and age range of the YSOs are estimated to be ~0.1-2 M ⊙ and 0.5-2 Myr using optical (V/V-I) and NIR (J/J-H) CM diagrams. The mean age of the YSOs is found to be ~1 Myr, which is of the order of dynamical age of 1.07 Myr of the H II region. Using the estimated range of visual extinction (1.1-25 mag) from literature and NIR data for the region, spectral energy distribution models have been implemented for selected YSOs which show masses and ages to be consistent with estimated values. The spatial distribution of YSOs shows an evolutionary sequence, suggesting triggered star formation in the region. The star formation seems to have propagated from the ionizing star toward the cold dark cloud LDN1657A located west of Sh2-297.

  12. STAR FORMATION ACTIVITY IN THE GALACTIC H II REGION Sh2-297

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Mallick, K. K.; Ojha, D. K.; Dewangan, L. K.

    We present a multiwavelength study of the Galactic H II region Sh2-297, located in the Canis Major OB1 complex. Optical spectroscopic observations are used to constrain the spectral type of ionizing star HD 53623 as B0V. The classical nature of this H II region is affirmed by the low values of electron density and emission measure, which are calculated to be 756 cm{sup -3} and 9.15 Multiplication-Sign 10{sup 5} cm{sup -6} pc using the radio continuum observations at 610 and 1280 MHz, and Very Large Array archival data at 1420 MHz. To understand local star formation, we identified the youngmore » stellar object (YSO) candidates in a region of area {approx}7.'5 Multiplication-Sign 7.'5 centered on Sh2-297 using grism slitless spectroscopy (to identify the H{alpha} emission line stars), and near infrared (NIR) observations. NIR YSO candidates are further classified into various evolutionary stages using color-color and color-magnitude (CM) diagrams, giving 50 red sources (H - K > 0.6) and 26 Class II-like sources. The mass and age range of the YSOs are estimated to be {approx}0.1-2 M {sub Sun} and 0.5-2 Myr using optical (V/V-I) and NIR (J/J-H) CM diagrams. The mean age of the YSOs is found to be {approx}1 Myr, which is of the order of dynamical age of 1.07 Myr of the H II region. Using the estimated range of visual extinction (1.1-25 mag) from literature and NIR data for the region, spectral energy distribution models have been implemented for selected YSOs which show masses and ages to be consistent with estimated values. The spatial distribution of YSOs shows an evolutionary sequence, suggesting triggered star formation in the region. The star formation seems to have propagated from the ionizing star toward the cold dark cloud LDN1657A located west of Sh2-297.« less

  13. Criteria for selecting a CO/sub 2//climate change region of study

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Edmonds, J.; Cushman, R.; Easterling, W.

    One of the most important research issues active today is the greenhouse issue. Progress has been made in exploring the relationship between human activities and the accumulation of CO/sub 2/ and other radiatively important gases in the atmosphere. While significant research remains in refining our understanding of the timing of possible CO/sub 2//climate change, the examination of the nature and magnitude of consequences of CO/sub 2//climate change remains in a relatively early stage of development. While the accumulation of greenhouse gases in the atmosphere may be a global problem, the consequences of CO/sub 2//climate change will be experienced regionally. Itmore » is therefore critical that methods be developed to address the regional examination of CO/sub 2//climate change. An analytical framework is described and a ''cookie cutter'' technique is utilized to deal with multiple resource sectors in selecting a Region of Study. The result leads to the selection of the four midwestern states of Kansas, Nebraska, Iowa, and Missouri. The role of information systems, uncertainty analysis, and knowledge transfer is discussed. 19 refs., 2 figs.« less

  14. Photoionization Models of the H_2 Emission of the Narrow Line Region of AGNs

    NASA Astrophysics Data System (ADS)

    Aleman, I.; Gruenwald, R.

    2011-05-01

    The excitation mechanism of the narrow line region (NLR) of AGNs is still an open question. Excitation by UV radiation from O and B stars, x-rays from the central black hole, shock from supernovae or jets, or a combination of these mechanisms have been suggested. In the present work, we use photoionization models to study the excitation mechanisms of the H_2 infrared emission lines in the NLR. In the literature, analyzes of the H_2 emission have been done assuming that the molecules is present only in neutral regions (photodissociation regions, x-ray-dominated regions, or shocks; Veilleux et al. 1997, Krabbe et al. 2000, Rigopoulou et al. 2002, Rodriguez-Ardila et al. 2004, 2005, and Davies et al. 2005). However, they are not conclusive. In previous work (Aleman & Gruenwald 2004, 2011), we show that the H_2 emission from the ionized region of PNe can be significant for planetary nebulae (PNe) with hot central stars (T⋆ > 150000 K). Such stars produce copious amounts of high energy photons, which create an extended partially ionized region that favors the H_2 survival. The conditions in the NLR are similar to those in PNe with hot central stars, so we can expect that the H_2 emission might also be important. We obtain and analyze a grid of photoionization models for different NRL parameters. We study the resulting H_2 density and emission, as well as, the formation, destruction, excitation, and de-excitation mechanisms. The higher values observed for the H_2 1-0 S(1)/Brγ ratio cannot be reproduced by our models. The calculated ratios are between 10^-8 and 10^-1, while the observational ration can be as high as 10. The calculated ratio is strongly anti-correlated with the ionization parameter (U) and only models with U<10-3 result in ratios inside the observational range. We show that the NLR is an environment more hostile to the H_2 molecule than the ionized region of PNe. Another interesting result of our calculations is that the H_2 formation on grain surfaces

  15. Development of a bead-based Luminex assay using lipopolysaccharide specific monoclonal antibodies to detect biological threats from Brucella species.

    PubMed

    Silbereisen, Angelika; Tamborrini, Marco; Wittwer, Matthias; Schürch, Nadia; Pluschke, Gerd

    2015-10-05

    Brucella, a Gram-negative bacterium, is classified as a potential bioterrorism agent mainly due to the low dose needed to cause infection and the ability to transmit the bacteria via aerosols. Goats/sheep, cattle, pigs, dogs, sheep and rodents are infected by B. melitensis, B. abortus, B. suis, B. canis, B. ovis and B. neotomae, respectively, the six classical Brucella species. Most human cases are caused by B. melitensis and B. abortus. Our aim was to specifically detect Brucellae with 'smooth' lipopolysaccharide (LPS) using a highly sensitive monoclonal antibody (mAb) based immunological assay. To complement molecular detection systems for potential bioterror agents, as required by international biodefense regulations, sets of mAbs were generated by B cell hybridoma technology and used to develop immunological assays. The combination of mAbs most suitable for an antigen capture assay format was identified and an immunoassay using the Luminex xMAP technology was developed. MAbs specific for the LPS O-antigen of Brucella spp. were generated by immunising mice with inactivated B. melitensis or B. abortus cells. Most mAbs recognised both B. melitensis and B. abortus and antigen binding was not impeded by inactivation of the bacterial cells by γ irradiation, formalin or heat treatment, a step required to analyse the samples immunologically under biosafety level two conditions. The Luminex assay recognised all tested Brucella species with 'smooth' LPS with detection limits of 2×10(2) to 8×10(4) cells per mL, depending on the species tested. Milk samples spiked with Brucella spp. cells were identified successfully using the Luminex assay. In addition, the bead-based immunoassay was integrated into a multiplex format, allowing for simultaneous, rapid and specific detection of Brucella spp., Bacillus anthracis, Francisella tularensis and Yersinia pestis within a single sample. Overall, the robust Luminex assay should allow detection of Brucella spp. in both natural

  16. The Chandra Source Catalog 2.0: the Galactic center region

    NASA Astrophysics Data System (ADS)

    Civano, Francesca Maria; Allen, Christopher E.; Anderson, Craig S.; Budynkiewicz, Jamie A.; Burke, Douglas; Chen, Judy C.; D'Abrusco, Raffaele; Doe, Stephen M.; Evans, Ian N.; Evans, Janet D.; Fabbiano, Giuseppina; Gibbs, Danny G., II; Glotfelty, Kenny J.; Graessle, Dale E.; Grier, John D.; Hain, Roger; Hall, Diane M.; Harbo, Peter N.; Houck, John C.; Lauer, Jennifer L.; Laurino, Omar; Lee, Nicholas P.; Martínez-Galarza, Juan Rafael; McCollough, Michael L.; McDowell, Jonathan C.; Miller, Joseph; McLaughlin, Warren; Morgan, Douglas L.; Mossman, Amy E.; Nguyen, Dan T.; Nichols, Joy S.; Nowak, Michael A.; Paxson, Charles; Plummer, David A.; Primini, Francis Anthony; Rots, Arnold H.; Siemiginowska, Aneta; Sundheim, Beth A.; Tibbetts, Michael; Van Stone, David W.; Zografou, Panagoula

    2018-01-01

    The second release of the Chandra Source Catalog (CSC 2.0) comprises all the 10,382 ACIS and HRC-I imaging observations taken by Chandra and released publicly through the end of 2014. Among these, 534 single observations surrounding the Galactic center are included, covering a total area of ~19deg2 and a total exposure time of ~9 Ms.The single 534 observations were merged into 379 stacks (overlapping observations with aim-points within 60") to increase the flux limit for source detection purposes.Thanks to the combination of the point source detection algorithm with the maximum likelihood technique used to asses the source significance, ~21,000 detections are listed in the CSC 2.0 for this field only, 80% of which are unique sources. The central region of this field around the SgrA* location has the deepest exposure of 2.2 Ms and the highest source density with ~5000 sources. In this poster, we present details about this region including source distribution and density, coverage, exposure.This work has been supported by NASA under contract NAS 8-03060 to the Smithsonian Astrophysical Observatory for operation of the ChandraX-ray Center.

  17. Intracellular bacteria interfere with dendritic cell functions: role of the type I interferon pathway.

    PubMed

    Gorvel, Laurent; Textoris, Julien; Banchereau, Romain; Ben Amara, Amira; Tantibhedhyangkul, Wiwit; von Bargen, Kristin; Ka, Mignane B; Capo, Christian; Ghigo, Eric; Gorvel, Jean-Pierre; Mege, Jean-Louis

    2014-01-01

    Dendritic cells (DCs) orchestrate host defenses against microorganisms. In infectious diseases due to intracellular bacteria, the inefficiency of the immune system to eradicate microorganisms has been attributed to the hijacking of DC functions. In this study, we selected intracellular bacterial pathogens with distinct lifestyles and explored the responses of monocyte-derived DCs (moDCs). Using lipopolysaccharide as a control, we found that Orientia tsutsugamushi, the causative agent of scrub typhus that survives in the cytosol of target cells, induced moDC maturation, as assessed by decreased endocytosis activity, the ability to induce lymphocyte proliferation and the membrane expression of phenotypic markers. In contrast, Coxiella burnetii, the agent of Q fever, and Brucella abortus, the agent of brucellosis, both of which reside in vacuolar compartments, only partly induced the maturation of moDCs, as demonstrated by a phenotypic analysis. To analyze the mechanisms used by C. burnetii and B. abortus to alter moDC activation, we performed microarray and found that C. burnetii and B. abortus induced a specific signature consisting of TLR4, TLR3, STAT1 and interferon response genes. These genes were down-modulated in response to C. burnetii and B. abortus but up-modulated in moDCs activated by lipopolysaccharide and O. tsutsugamushi. This transcriptional alteration was associated with the defective interferon-β production. This study demonstrates that intracellular bacteria specifically affect moDC responses and emphasizes how C. burnetii and B. abortus interfere with moDC activation and the antimicrobial immune response. We believe that comparing infection by several bacterial species may be useful for defining new pathways and biomarkers and for developing new treatment strategies.

  18. Whole-genome analyses of speciation events in pathogenic Brucellae

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Chain, Patrick S. G.; Comerci, Diego J.; Tolmasky, Marcelo E.

    Despite their high DNA identity and a proposal to group classical Brucella species as biovars of Brucella melitensis, the commonly recognized Brucella species can be distinguished by distinct biochemical and fatty acid characters, as well as by a marked host range (e.g., Brucella suis for swine, B. melitensis for sheep and goats, and Brucella abortus for cattle). Here we present the genome of B. abortus 2308, the virulent prototype biovar 1 strain, and its comparison to the two other human pathogenic Brucella species and to B. abortus field isolate 9-941. The global distribution of pseudogenes, deletions, and insertions supports previousmore » indications that B. abortus and B. melitensis share a common ancestor that diverged from B. suis. With the exception of a dozen genes, the genetic complements of both B. abortus strains are identical, whereas the three species differ in gene content and pseudogenes. The pattern of species-specific gene inactivations affecting transcriptional regulators and outer membrane proteins suggests that these inactivations may play an important role in the establishment of host specificity and may have been a primary driver of speciation in the genus Brucella. Despite being nonmotile, the brucellae contain flagellum gene clusters and display species-specific flagellar gene inactivations, which lead to the putative generation of different versions of flagellum-derived structures and may contribute to differences in host specificity and virulence. Metabolic changes such as the lack of complete metabolic pathways for the synthesis of numerous compounds (e.g., glycogen, biotin, NAD, and choline) are consistent with adaptation of brucellae to an intracellular life-style.« less

  19. Whole-genome analyses of the speciation events in the pathogenic Brucellae

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Chain, P; Comerci, D; Tolmasky, M

    Despite their high DNA identity and a proposal to group classical Brucella species as biovars of B. melitensis, the commonly recognized Brucella species can be distinguished by distinct biochemical and fatty acid characters as well as by a marked host range (e.g. B. suis for swine, B. melitensis for sheep and goats, B. abortus for cattle). Here we present the genome of B. abortus 2308, the virulent prototype biovar 1 strain, and its comparison to the two other human pathogenic Brucellae species and to the B. abortus field isolate 9-941. The global distribution of pseudogenes, deletions and insertions support previousmore » indications that B. abortus and B. melitensis share a common ancestor that diverged from B. suis. With the exception of a dozen genes, the genetic complement of both B. abortus strains is identical, whereas the three species differ in gene content and pseudogenes. The pattern of species-specific gene inactivations affecting transcriptional regulators and outer membrane proteins suggest that these inactivations may play an important role in the establishment of host-specificity and may have been a primary driver of speciation in the Brucellae. Despite being non-motile, the Brucellae contain flagellum gene clusters and display species-specific flagellar gene inactivations, which lead to the putative generation of different versions of flagellum-derived structures, and may contribute to differences in host-specificity and virulence. Metabolic changes such as the lack of complete metabolic pathways for the synthesis of numerous compounds (e.g. glycogen, biotin, NAD, and choline) are consistent with adaptation of Brucellae to an intracellular lifestyle.« less

  20. Common regions of deletion in chromosome regions 3p12 and 3p14.2 in primary clear cell renal carcinomas.

    PubMed

    Lubinski, J; Hadaczek, P; Podolski, J; Toloczko, A; Sikorski, A; McCue, P; Druck, T; Huebner, K

    1994-07-15

    Nearly all clear cell renal cell carcinomas (RCCs) exhibit loss of alleles on the short arm of chromosome 3. Loss and mutation at the von Hippel-Lindau (VHL) gene at 3p25 probably occurs in most RCCs and, since the VHL gene was recently cloned, data on VHL involvement in RCCs is accumulating. However, the region 3p14-p12, a region that contains the familial RCC-associated t(3;8)(p14.2;q24) chromosome translocation and the small cell lung carcinoma-associated homozygous deletion at 3p13-12, has also been reported to exhibit allele loss in a large fraction of RCCs. In order to focus future studies on potential suppressor genes in the 3p14-p12 region, we have studied allele loss in 30 RCCs with 9 polymorphic simple sequence repeat markers spanning 3p21.1-p12. Partial losses in the 3p21-p12 region were observed, allowing determination of common regions of loss of heterozygosity overlap in 15 RCCs. Results suggested that most RCCs exhibit loss in a region which brackets the t(3;8) familial chromosome translocation at 3p14.2, and some show additional deletions within the U2020 small cell lung carcinoma deletion at 3p12.

  1. Long-term Variability of H2CO Masers in Star-forming Regions

    NASA Astrophysics Data System (ADS)

    Andreev, N.; Araya, E. D.; Hoffman, I. M.; Hofner, P.; Kurtz, S.; Linz, H.; Olmi, L.; Lorran-Costa, I.

    2017-10-01

    We present results of a multi-epoch monitoring program on variability of 6 cm formaldehyde (H2CO) masers in the massive star-forming region NGC 7538 IRS 1 from 2008 to 2015, conducted with the Green Bank Telescope, the Westerbork Radio Telescope , and the Very Large Array. We found that the similar variability behaviors of the two formaldehyde maser velocity components in NGC 7538 IRS 1 (which was pointed out by Araya and collaborators in 2007) have continued. The possibility that the variability is caused by changes in the maser amplification path in regions with similar morphology and kinematics is discussed. We also observed 12.2 GHz methanol and 22.2 GHz water masers toward NGC 7538 IRS 1. The brightest maser components of CH3OH and H2O species show a decrease in flux density as a function of time. The brightest H2CO maser component also shows a decrease in flux density and has a similar LSR velocity to the brightest H2O and 12.2 GHz CH3OH masers. The line parameters of radio recombination lines and the 20.17 and 20.97 GHz CH3OH transitions in NGC 7538 IRS 1 are also reported. In addition, we observed five other 6 cm formaldehyde maser regions. We found no evidence of significant variability of the 6 cm masers in these regions with respect to previous observations, the only possible exception being the maser in G29.96-0.02. All six sources were also observed in the {{{H}}}213{CO} isotopologue transition of the 6 cm H2CO line; {{{H}}}213{CO} absorption was detected in five of the sources. Estimated column density ratios [{{{H}}}212{CO}]/[{{{H}}}213{CO}] are reported.

  2. Regional uncertainty of GOSAT XCO2 retrievals in China: quantification and attribution

    NASA Astrophysics Data System (ADS)

    Bie, Nian; Lei, Liping; Zeng, ZhaoCheng; Cai, Bofeng; Yang, Shaoyuan; He, Zhonghua; Wu, Changjiang; Nassar, Ray

    2018-03-01

    The regional uncertainty of the column-averaged dry air mole fraction of CO2 (XCO2) retrieved using different algorithms from the Greenhouse gases Observing SATellite (GOSAT) and its attribution are still not well understood. This paper investigates the regional performance of XCO2 within a latitude band of 37-42° N segmented into 8 cells in a grid of 5° from west to east (80-120° E) in China, where typical land surface types and geographic conditions exist. The former includes desert, grassland and built-up areas mixed with cropland; and the latter includes anthropogenic emissions that change from small to large from west to east, including those from the megacity of Beijing. For these specific cells, we evaluate the regional uncertainty of GOSAT XCO2 retrievals by quantifying and attributing the consistency of XCO2 retrievals from four algorithms (ACOS, NIES, OCFP and SRFP) by intercomparison. These retrievals are then specifically compared with simulated XCO2 from the high-resolution nested model in East Asia of the Goddard Earth Observing System 3-D chemical transport model (GEOS-Chem). We also introduce the anthropogenic CO2 emissions data generated from the investigation of surface emitting point sources that was conducted by the Ministry of Environmental Protection of China to GEOS-Chem simulations of XCO2 over the Chinese mainland. The results indicate that (1) regionally, the four algorithms demonstrate smaller absolute biases of 0.7-1.1 ppm in eastern cells, which are covered by built-up areas mixed with cropland with intensive anthropogenic emissions, than those in the western desert cells (1.0-1.6 ppm) with a high-brightness surface from the pairwise comparison results of XCO2 retrievals. (2) Compared with XCO2 simulated by GEOS-Chem (GEOS-XCO2), the XCO2 values from ACOS and SRFP have better agreement, while values from OCFP are the least consistent with GEOS-XCO2. (3) Viewing attributions of XCO2 in the spatio-temporal pattern, ACOS and SRFP

  3. The environment and star formation of H II region Sh2-163: a multi-wavelength study

    NASA Astrophysics Data System (ADS)

    Yu, Naiping; Wang, Jun-Jie; Li, Nan

    2014-12-01

    To investigate the environment of H II region Sh2-163 and search for evidence of triggered star formation in this region, we performed a multi-wavelength study of this H II region. Most of our data were taken from large-scale surveys: 2MASS, CGPS, MSX and SCUBA. We also made CO molecular line observations, using the 13.7-m telescope. The ionized region of Sh2-163 is detected by both the optical and radio continuum observations. Sh2-163 is partially bordered by an arc-like photodissociation region (PDR), which is coincident with the strongest optical and radio emissions, indicating interactions between the H II region and the surrounding interstellar medium. Two molecular clouds were discovered on the border of the PDR. The morphology of these two clouds suggests they are compressed by the expansion of Sh2-163. In cloud A, we found two molecular clumps. And it seems star formation in clump A2 is much more active than in clump A1. In cloud B, we found new outflow activities and massive star(s) are forming inside. Using 2MASS photometry, we tried to search for embedded young stellar object (YSO) candidates in this region. The very good agreement between CO emission, infrared shell and YSOs suggest that it is probably a star formation region triggered by the expansion of Sh2-163. We also found the most likely massive protostar related to IRAS 23314+6033.

  4. 2-D Density and Directional Analysis of Fault Systems in the Zagros Region (Iran) on a Regional Scale

    NASA Astrophysics Data System (ADS)

    Hashemi, Seyed Naser; Baizidi, Chavare

    2018-04-01

    In this paper, 2-D spatial variation of the frequency and length density and frequency-length relation of large-scale faults in the Zagros region (Iran), as a typical fold-and-thrust belt, were examined. Moreover, the directional analysis of these faults as well as the scale dependence of the orientations was studied. For this purpose, a number of about 8000 faults with L ≥ 1.0 km were extracted from the geological maps covering the region, and then, the data sets were analyzed. The overall pattern of the frequency/length distribution of the total faults of the region acceptably fits with a power-law relation with exponent 1.40, with an obvious change in the gradient in L = 12.0 km. In addition, maps showing the spatial variation of fault densities over the region indicate that the maximum values of the frequency and length density of the faults are attributed to the northeastern part of the region and parallel to the suture zone, respectively, and the fault density increases towards the central parts of the belt. Moreover, the directional analysis of the fault trends gives a dominant preferred orientation trend of 300°-330° and the assessment of the scale dependence of the fault directions demonstrates that larger faults show higher degrees of preferred orientations. As a result, it is concluded that the evolutionary path of the faulting process in this region can be explained by increasing the number of faults rather than the growth in the fault lengths and also it seems that the regional-scale faults in this region are generated by a nearly steady-state tectonic stress regime.

  5. Loss of imprinting at the Dlk1-Gtl2 locus caused by insertional mutagenesis in the Gtl2 5' region

    PubMed Central

    Steshina, Ekaterina Y; Carr, Michael S; Glick, Elena A; Yevtodiyenko, Aleksey; Appelbe, Oliver K; Schmidt, Jennifer V

    2006-01-01

    Background The Dlk1 and Gtl2 genes define a region of mouse chromosome 12 that is subject to genomic imprinting, the parental allele-specific expression of a gene. Although imprinted genes play important roles in growth and development, the mechanisms by which imprinting is established and maintained are poorly understood. Differentially methylated regions (DMRs), which carry methylation on only one parental allele, are involved in imprinting control at many loci. The Dlk1-Gtl2 region contains three known DMRs, the Dlk1 DMR in the 3' region of Dlk1, the intergenic DMR 15 kb upstream of Gtl2, and the Gtl2 DMR at the Gtl2 promoter. Three mouse models are analyzed here that provide new information about the regulation of Dlk1-Gtl2 imprinting. Results A previously existing insertional mutation (Gtl2lacZ), and a targeted deletion in which the Gtl2 upstream region was replaced by a Neo cassette (Gtl2Δ5'Neo), display partial lethality and dwarfism upon paternal inheritance. Molecular characterization shows that both mutations cause loss of imprinting and changes in expression of the Dlk1, Gtl2 and Meg8/Rian genes. Dlk1 levels are decreased upon paternal inheritance of either mutation, suggesting Dlk1 may be causative for the lethality and dwarfism. Loss of imprinting on the paternal chromosome in both Gtl2lacZ and Gtl2Δ5'Neo mice is accompanied by the loss of paternal-specific Gtl2 DMR methylation, while maternal loss of imprinting suggests a previously unknown regulatory role for the maternal Gtl2 DMR. Unexpectedly, when the Neo gene is excised, Gtl2Δ5' animals are of normal size, imprinting is unchanged and the Gtl2 DMR is properly methylated. The exogenous DNA sequences integrated upstream of Gtl2 are therefore responsible for the growth and imprinting effects. Conclusion These data provide further evidence for the coregulation of the imprinted Dlk1 and Gtl2 genes, and support a role for Dlk1 as an important neonatal growth factor. The ability of the Gtl2lacZ and

  6. Two-step transition towards the reversibility region in Bi2Sr2CaCu2O8-δ single crystals

    NASA Astrophysics Data System (ADS)

    Pastoriza, H.; de La Cruz, F.; Mitzi, D. B.; Kapitulnik, A.

    1992-10-01

    We have performed magnetization measurements on Bi2Sr2CaCu2O8-δ single crystals in the c^ crystallographic direction for fields from 2 Oe up to 700 Oe. The results strongly suggest that the reversible thermodynamic region is achieved after the vortex flux structure shows an abrupt transition at a temperature lower than that determined by the irreversibility line.

  7. Phytochemical Characteristics, Free Radical Scavenging Activities, and Neuroprotection of Five Medicinal Plant Extracts

    PubMed Central

    Chang, Chia Lin; Lin, Che San; Lai, Guia Hung

    2012-01-01

    The objective of this study was to determine phytochemical characteristics, chemiluminescence antioxidant capacities, and neuroprotective effects on PC12 cells for methanol extracts of Spatholobus suberectus, Uncaria rhynchophylla, Alpinia officinarum, Drynaria fortunei, and Crataegus pinnatifida. The C. pinnatifida extract (CPE) afforded the greatest yield and total phenolic content. The S. suberectus extract (SSE) yielded the greatest total flavonoid content. The U. rhynchophylla extract (URE) produced the greatest total tannin content, and the A. officinarum extract (AOE) produced the greatest total triterpenoid content. The D. fortunei extract, assayed using horseradish peroxidase-luminol-hydrogen peroxide (H2O2), and AOE using pyrogallol-luminol assay each exhibited better antioxidant activity than the L-ascorbic acid and 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid did. The CPE, SSE, and URE presented neurogrowth effects and neuroprotective activities on H2O2-induced PC12 cell death at 0.5–5.0 μg/mL. The CPE represents a promising medicinal plant source for the treatment of H2O2-induced neurodegenerative disease, because of its useful phytochemical characteristics. PMID:21845204

  8. Use of ITS2 region as the universal DNA barcode for plants and animals.

    PubMed

    Yao, Hui; Song, Jingyuan; Liu, Chang; Luo, Kun; Han, Jianping; Li, Ying; Pang, Xiaohui; Xu, Hongxi; Zhu, Yingjie; Xiao, Peigen; Chen, Shilin

    2010-10-01

    The internal transcribed spacer 2 (ITS2) region of nuclear ribosomal DNA is regarded as one of the candidate DNA barcodes because it possesses a number of valuable characteristics, such as the availability of conserved regions for designing universal primers, the ease of its amplification, and sufficient variability to distinguish even closely related species. However, a general analysis of its ability to discriminate species in a comprehensive sample set is lacking. In the current study, 50,790 plant and 12,221 animal ITS2 sequences downloaded from GenBank were evaluated according to sequence length, GC content, intra- and inter-specific divergence, and efficiency of identification. The results show that the inter-specific divergence of congeneric species in plants and animals was greater than its corresponding intra-specific variations. The success rates for using the ITS2 region to identify dicotyledons, monocotyledons, gymnosperms, ferns, mosses, and animals were 76.1%, 74.2%, 67.1%, 88.1%, 77.4%, and 91.7% at the species level, respectively. The ITS2 region unveiled a different ability to identify closely related species within different families and genera. The secondary structure of the ITS2 region could provide useful information for species identification and could be considered as a molecular morphological characteristic. As one of the most popular phylogenetic markers for eukaryota, we propose that the ITS2 locus should be used as a universal DNA barcode for identifying plant species and as a complementary locus for CO1 to identify animal species. We have also developed a web application to facilitate ITS2-based cross-kingdom species identification (http://its2-plantidit.dnsalias.org).

  9. Use of ITS2 Region as the Universal DNA Barcode for Plants and Animals

    PubMed Central

    Luo, Kun; Han, Jianping; Li, Ying; Pang, Xiaohui; Xu, Hongxi; Zhu, Yingjie; Xiao, Peigen; Chen, Shilin

    2010-01-01

    Background The internal transcribed spacer 2 (ITS2) region of nuclear ribosomal DNA is regarded as one of the candidate DNA barcodes because it possesses a number of valuable characteristics, such as the availability of conserved regions for designing universal primers, the ease of its amplification, and sufficient variability to distinguish even closely related species. However, a general analysis of its ability to discriminate species in a comprehensive sample set is lacking. Methodology/Principal Findings In the current study, 50,790 plant and 12,221 animal ITS2 sequences downloaded from GenBank were evaluated according to sequence length, GC content, intra- and inter-specific divergence, and efficiency of identification. The results show that the inter-specific divergence of congeneric species in plants and animals was greater than its corresponding intra-specific variations. The success rates for using the ITS2 region to identify dicotyledons, monocotyledons, gymnosperms, ferns, mosses, and animals were 76.1%, 74.2%, 67.1%, 88.1%, 77.4%, and 91.7% at the species level, respectively. The ITS2 region unveiled a different ability to identify closely related species within different families and genera. The secondary structure of the ITS2 region could provide useful information for species identification and could be considered as a molecular morphological characteristic. Conclusions/Significance As one of the most popular phylogenetic markers for eukaryota, we propose that the ITS2 locus should be used as a universal DNA barcode for identifying plant species and as a complementary locus for CO1 to identify animal species. We have also developed a web application to facilitate ITS2-based cross-kingdom species identification (http://its2-plantidit.dnsalias.org). PMID:20957043

  10. Domination Problem for Vector Measures and Applications to Nonstationary Processes.

    DTIC Science & Technology

    1981-09-23

    meas- ures, which have p-semi-variation finite, I a p a 2 . The work here do- pieda in part on an inequality of Grothendieck- Pietsch . The rest of the...7], Corol. 2 to Thi. 4.3 and Prop. 3.1, the Latter is the Grothendieck- Pietsch inequality alluded to in the Introduction), the space I - C(S) being

  11. Anti-genotoxicity of trans-anethole and eugenol in mice.

    PubMed

    Abraham, S K

    2001-05-01

    The naturally occurring flavouring agents trans-anethole and eugenol were evaluated for antigenotoxic effects in mice. The test doses of trans-anethole (40-400 mg/kg body weight) and eugenol (50-500 mg/kg weight) were administered by gavage 2 and 20 h before the genotoxins were injected intraperitoneally. Anti-genotoxic effects were assessed in the mouse bone marrow micronucleus test. Pretreatment with trans-anethole and eugenol led to significant antigenotoxic effects against cyclophosphamide (CPH), procarbazine (PCB), N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) and urethane (URE). In addition, trans-anethole inhibited the genotoxicity of ethyl methane sulfonate (EMS). Both trans-anethole and eugenol exerted dose-related antigenotoxic effects against PCB and URE. There was no significant increase in genotoxicity when trans-anethole (40-400 mg/kg body weight) and eugenol (50-500 mg/kg body weight) were administered alone.

  12. Exploring the Dynamics of Directed Studies Courses: Student, Instructor, and Administrator Perspectives

    ERIC Educational Resources Information Center

    Hvenegaard, Glen; Link, Anne-Marie L.; Moore, Sean E.; Wesselius, Janet C.

    2013-01-01

    North American universities are encouraged to increase opportunities for undergraduate research experiences (UREs). To this end, many universities offer directed studies courses (DSCs) which are 1-2 semester long courses involving one-on-one instruction, with a focus on student-led independent research. Building on the understanding of dynamics…

  13. Methylation Analysis of the BMPR2 Gene Promoter Region in Patients With Pulmonary Arterial Hypertension.

    PubMed

    Pousada, Guillermo; Baloira, Adolfo; Valverde, Diana

    2016-06-01

    Pulmonary arterial hypertension is characterizated by obstruction of the pulmonary arteries. The gene mainly related to pathology is the bone morphogenetic protein receptor type II (BMPR2). The aim of this study was to analyze the methylation pattern of the BMPR2 promoter region in patients and controls. We used Methyl Primer Express(®) v.1.0 and MatInspector softwares to analyze this region. Genomic DNA obtained from the peripheral blood of patients and controls was modified with sodium bisulphite. Methylation was analyzed using methylation-specific PCR. DNA treated with CpG methyltransferase was used as a positive control for methylation and H1299 cell culture DNA was used as positive control for gene expression. We identified a CpG island, which may have been methylated, in the BMPR2 promoter region, in addition to NIT-2 (global-acting regulatory protein), sex-determining region Y) and heat shock factor transcription factor binding sites. We found no evidence of methylation in patients and controls. No methylated CpG sites were identified in H1299 cells expressing the BMPR2 gene. The BMPR2 promoter region is the most suitable for study because of the high number of transcription factor binding sites that could alter gene function. No evidence of methylation was detected in this region in patients and controls. Copyright © 2015 SEPAR. Published by Elsevier Espana. All rights reserved.

  14. Detecting regional patterns of changing CO2 flux in Alaska

    PubMed Central

    Parazoo, Nicholas C.; Wofsy, Steven C.; Koven, Charles D.; Sweeney, Colm; Lawrence, David M.; Lindaas, Jakob; Chang, Rachel Y.-W.; Miller, Charles E.

    2016-01-01

    With rapid changes in climate and the seasonal amplitude of carbon dioxide (CO2) in the Arctic, it is critical that we detect and quantify the underlying processes controlling the changing amplitude of CO2 to better predict carbon cycle feedbacks in the Arctic climate system. We use satellite and airborne observations of atmospheric CO2 with climatically forced CO2 flux simulations to assess the detectability of Alaskan carbon cycle signals as future warming evolves. We find that current satellite remote sensing technologies can detect changing uptake accurately during the growing season but lack sufficient cold season coverage and near-surface sensitivity to constrain annual carbon balance changes at regional scale. Airborne strategies that target regular vertical profile measurements within continental interiors are more sensitive to regional flux deeper into the cold season but currently lack sufficient spatial coverage throughout the entire cold season. Thus, the current CO2 observing network is unlikely to detect potentially large CO2 sources associated with deep permafrost thaw and cold season respiration expected over the next 50 y. Although continuity of current observations is vital, strategies and technologies focused on cold season measurements (active remote sensing, aircraft, and tall towers) and systematic sampling of vertical profiles across continental interiors over the full annual cycle are required to detect the onset of carbon release from thawing permafrost. PMID:27354511

  15. Detecting regional patterns of changing CO 2 flux in Alaska

    DOE PAGES

    Parazoo, Nicholas C.; Commane, Roisin; Wofsy, Steven C.; ...

    2016-06-27

    With rapid changes in climate and the seasonal amplitude of carbon dioxide (CO 2) in the Arctic, it is critical that we detect and quantify the underlying processes controlling the changing amplitude of CO 2 to better predict carbon cycle feedbacks in the Arctic climate system. We use satellite and airborne observations of atmospheric CO 2 with climatically forced CO 2 flux simulations to assess the detectability of Alaskan carbon cycle signals as future warming evolves. We find that current satellite remote sensing technologies can detect changing uptake accurately during the growing season but lack sufficient cold season coverage andmore » near-surface sensitivity to constrain annual carbon balance changes at regional scale. Airborne strategies that target regular vertical profile measurements within continental interiors are more sensitive to regional flux deeper into the cold season but currently lack sufficient spatial coverage throughout the entire cold season. Thus, the current CO 2 observing network is unlikely to detect potentially large CO 2 sources associated with deep permafrost thaw and cold season respiration expected over the next 50 y. In conclusion, although continuity of current observations is vital, strategies and technologies focused on cold season measurements (active remote sensing, aircraft, and tall towers) and systematic sampling of vertical profiles across continental interiors over the full annual cycle are required to detect the onset of carbon release from thawing permafrost.« less

  16. Estimating regional CO2 and CH4 fluxes using GOSAT XCO2 and XCH4 observations

    NASA Astrophysics Data System (ADS)

    Fraser, A. C.; Palmer, P. I.; Feng, L.; Parker, R.; Boesch, H.; Cogan, A. J.

    2012-12-01

    We infer regional monthly surface flux estimates for CO2 and CH4, June 2009-December 2010, from proxy dry-air column-averaged mole fractions of CO2 and CH4 from the Greenhouse gases Observing SATellite (GOSAT) using an ensemble Kalman Filter combined with the GEOS-Chem chemistry transport model. We compare these flux estimates with estimates inferred from in situ surface mole fraction measurements and from combining in situ and GOSAT measurements in order to quantify the added value of GOSAT data above the conventional surface measurement network. We find that the error reduction, a measure of how much the posterior fluxes are being informed by the assimilated data, at least doubles when GOSAT measurements are used versus the surface only inversions, with the exception of regions that are well covered by the surface network at the spatial and temporal resolution of our flux estimation calculation. We have incorporated a new online bias correction scheme to account for GOSAT biases. We report global and regional flux estimates inferred from GOSAT and/or in situ measurements. While the global posterior fluxes from GOSAT and in situ measurements agree, we find significant differences in the regional fluxes, particularly over the tropics. We evaluate the posterior fluxes by comparing them against independent surface mole fraction, column, and aircraft measurements using the GEOS-Chem model as an intermediary.

  17. Immune Responses and Protection against Experimental Brucella suis biovar 1 Challenge in Non-vaccinated or RB51-Vaccinated Cattle

    USDA-ARS?s Scientific Manuscript database

    Twenty Hereford heifers, approximately 9 months of age, were vaccinated with saline (control) or 2 x 10**10 CFU of Brucella abortus strain RB51 (RB51) vaccine. Immunologic responses after inoculation demonstrated significantly greater (P<0.05) antibody and proliferative responses to RB51 antigens i...

  18. Brucella and Osteoarticular Cell Activation: Partners in Crime

    PubMed Central

    Giambartolomei, Guillermo H.; Arriola Benitez, Paula C.; Delpino, M. Victoria

    2017-01-01

    Osteoarticular brucellosis is the most common presentation of human active disease although its prevalence varies widely. The three most common forms of osteoarticular involvement are sacroiliitis, spondylitis, and peripheral arthritis. The molecular mechanisms implicated in bone damage have been recently elucidated. B. abortus induces bone damage through diverse mechanisms in which TNF-α and the receptor activator of nuclear factor kappa-B ligand (RANKL)-the natural modulator of bone homeostasis are involved. These processes are driven by inflammatory cells, like monocytes/macrophages, neutrophils, Th17 CD4+ T, and B cells. In addition, Brucella abortus has a direct effect on osteoarticular cells and tilts homeostatic bone remodeling. These bacteria inhibit bone matrix deposition by osteoblasts (the only bone cells involved in bone deposition), and modify the phenotype of these cells to produce matrix metalloproteinases (MMPs) and cytokine secretion, contributing to bone matrix degradation. B. abortus also affects osteoclasts (cells naturally involved in bone resorption) by inducing an increase in osteoclastogenesis and osteoclast activation; thus, increasing mineral and organic bone matrix resorption, contributing to bone damage. Given that the pathology induced by Brucella species involved joint tissue, experiments conducted on synoviocytes revealed that besides inducing the activation of these cells to secrete chemokines, proinflammatory cytokines and MMPS, the infection also inhibits synoviocyte apoptosis. Brucella is an intracellular bacterium that replicates preferentially in the endoplasmic reticulum of macrophages. The analysis of B. abortus-infected synoviocytes indicated that bacteria also replicate in their reticulum suggesting that they could use this cell type for intracellular replication during the osteoarticular localization of the disease. Finally, the molecular mechanisms of osteoarticular brucellosis discovered recently shed light on how the

  19. The HOOK region of voltage-gated Ca2+ channel β subunits senses and transmits PIP2 signals to the gate.

    PubMed

    Park, Cheon-Gyu; Park, Yongsoo; Suh, Byung-Chang

    2017-02-01

    The β subunit of voltage-gated Ca 2+ (Ca V ) channels plays an important role in regulating gating of the α1 pore-forming subunit and its regulation by phosphatidylinositol 4,5-bisphosphate (PIP 2 ). Subcellular localization of the Ca V β subunit is critical for this effect; N-terminal-dependent membrane targeting of the β subunit slows inactivation and decreases PIP 2 sensitivity. Here, we provide evidence that the HOOK region of the β subunit plays an important role in the regulation of Ca V biophysics. Based on amino acid composition, we broadly divide the HOOK region into three domains: S (polyserine), A (polyacidic), and B (polybasic). We show that a β subunit containing only its A domain in the HOOK region increases inactivation kinetics and channel inhibition by PIP 2 depletion, whereas a β subunit with only a B domain decreases these responses. When both the A and B domains are deleted, or when the entire HOOK region is deleted, the responses are elevated. Using a peptide-to-liposome binding assay and confocal microscopy, we find that the B domain of the HOOK region directly interacts with anionic phospholipids via polybasic and two hydrophobic Phe residues. The β2c-short subunit, which lacks an A domain and contains fewer basic amino acids and no Phe residues in the B domain, neither associates with phospholipids nor affects channel gating dynamically. Together, our data suggest that the flexible HOOK region of the β subunit acts as an important regulator of Ca V channel gating via dynamic electrostatic and hydrophobic interaction with the plasma membrane. © 2017 Park et al.

  20. An unbiased risk estimator for image denoising in the presence of mixed poisson-gaussian noise.

    PubMed

    Le Montagner, Yoann; Angelini, Elsa D; Olivo-Marin, Jean-Christophe

    2014-03-01

    The behavior and performance of denoising algorithms are governed by one or several parameters, whose optimal settings depend on the content of the processed image and the characteristics of the noise, and are generally designed to minimize the mean squared error (MSE) between the denoised image returned by the algorithm and a virtual ground truth. In this paper, we introduce a new Poisson-Gaussian unbiased risk estimator (PG-URE) of the MSE applicable to a mixed Poisson-Gaussian noise model that unifies the widely used Gaussian and Poisson noise models in fluorescence bioimaging applications. We propose a stochastic methodology to evaluate this estimator in the case when little is known about the internal machinery of the considered denoising algorithm, and we analyze both theoretically and empirically the characteristics of the PG-URE estimator. Finally, we evaluate the PG-URE-driven parametrization for three standard denoising algorithms, with and without variance stabilizing transforms, and different characteristics of the Poisson-Gaussian noise mixture.

  1. Mechanisms of molecular transport through the urea channel of Helicobacter pylori

    PubMed Central

    McNulty, Reginald; Ulmschneider, Jakob P.; Luecke, Hartmut; Ulmschneider, Martin B.

    2013-01-01

    Helicobacter pylori survival in acidic environments relies on cytoplasmic hydrolysis of gastric urea into ammonia and carbon dioxide, which buffer the pathogen’s periplasm. Urea uptake is greatly enhanced and regulated by HpUreI, a proton-gated inner membrane channel protein essential for gastric survival of H. pylori. The crystal structure of HpUreI describes a static snapshot of the channel with two constriction sites near the center of the bilayer that are too narrow to allow passage of urea or even water. Here we describe the urea transport mechanism at atomic resolution, revealed by unrestrained microsecond equilibrium molecular dynamics simulations of the hexameric channel assembly. Two consecutive constrictions open to allow conduction of urea, which is guided through the channel by interplay between conserved residues that determine proton rejection and solute selectivity. Remarkably, HpUreI conducts water at rates equivalent to aquaporins, which might be essential for efficient transport of urea at small concentration gradients. PMID:24305683

  2. Mechanisms of molecular transport through the urea channel of Helicobacter pylori

    NASA Astrophysics Data System (ADS)

    McNulty, Reginald; Ulmschneider, Jakob P.; Luecke, Hartmut; Ulmschneider, Martin B.

    2013-12-01

    Helicobacter pylori survival in acidic environments relies on cytoplasmic hydrolysis of gastric urea into ammonia and carbon dioxide, which buffer the pathogen’s periplasm. Urea uptake is greatly enhanced and regulated by HpUreI, a proton-gated inner membrane channel protein essential for gastric survival of H. pylori. The crystal structure of HpUreI describes a static snapshot of the channel with two constriction sites near the center of the bilayer that are too narrow to allow passage of urea or even water. Here we describe the urea transport mechanism at atomic resolution, revealed by unrestrained microsecond equilibrium molecular dynamics simulations of the hexameric channel assembly. Two consecutive constrictions open to allow conduction of urea, which is guided through the channel by interplay between conserved residues that determine proton rejection and solute selectivity. Remarkably, HpUreI conducts water at rates equivalent to aquaporins, which might be essential for efficient transport of urea at small concentration gradients.

  3. Improving Project Performance through Implementation of Agile Methodologies in the Renewable Energy Construction Industry

    NASA Astrophysics Data System (ADS)

    Hernandez Mendez, Arturo

    Collaborative inquiry within undergraduate research experiences (UREs) is an effective curriculum tool to support student growth. This study seeks to understand how collaborative inquiry within undergraduate biology student experiences are affected within faculty mentored experiences and non-mentored experiences at a large private southeastern university. Undergraduate biology students engaged in UREs (faculty as mentor and non-mentor experiences) were examined for statistically significant differences in student self-efficacy. Self-efficacy was measured in three subcomponents (thinking and working like a scientist, scientific self-efficacy, and scientific identity) from student responses obtained in an online survey. Responses were analyzed using a nonparametric equivalent of a t test (Mann Whitney U test) to make comparisons between faculty mentored and non-mentored student groups. The conclusions of this study highlight the statistically significant effect of faculty mentoring in all three subcomponents. Faculty and university policy makers can apply these findings to develop further support for effective faculty mentoring practices in UREs.

  4. Managing Ocean Dumping in EPA Region 2

    EPA Pesticide Factsheets

    Overview of ocean dumping in NY, NJ and Puerto Rico. Includes materials dumped in the Region, ocean dumping permits issues, dredged material testing guidance, ocean disposal site descriptions and information, regional dredging teams and other partnerships

  5. Numerical investigation for the impact of CO2 geologic sequestration on regional groundwater flow

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Yamamoto, H.; Zhang, K.; Karasaki, K.

    Large-scale storage of carbon dioxide in saline aquifers may cause considerable pressure perturbation and brine migration in deep rock formations, which may have a significant influence on the regional groundwater system. With the help of parallel computing techniques, we conducted a comprehensive, large-scale numerical simulation of CO{sub 2} geologic storage that predicts not only CO{sub 2} migration, but also its impact on regional groundwater flow. As a case study, a hypothetical industrial-scale CO{sub 2} injection in Tokyo Bay, which is surrounded by the most heavily industrialized area in Japan, was considered, and the impact of CO{sub 2} injection on near-surfacemore » aquifers was investigated, assuming relatively high seal-layer permeability (higher than 10 microdarcy). A regional hydrogeological model with an area of about 60 km x 70 km around Tokyo Bay was discretized into about 10 million gridblocks. To solve the high-resolution model efficiently, we used a parallelized multiphase flow simulator TOUGH2-MP/ECO2N on a world-class high performance supercomputer in Japan, the Earth Simulator. In this simulation, CO{sub 2} was injected into a storage aquifer at about 1 km depth under Tokyo Bay from 10 wells, at a total rate of 10 million tons/year for 100 years. Through the model, we can examine regional groundwater pressure buildup and groundwater migration to the land surface. The results suggest that even if containment of CO{sub 2} plume is ensured, pressure buildup on the order of a few bars can occur in the shallow confined aquifers over extensive regions, including urban inlands.« less

  6. Why Fix N2 in High N Supply Regions?

    NASA Astrophysics Data System (ADS)

    Landolfi, A.; Koeve, W.; Oschlies, A.

    2016-02-01

    Growing slowly, marine N2 fixers are expected to be competitive where nitrogen (N) supply falls short relative to that of phosphorus (P) with respect to the cellular N:P ratio (R) of non-fixing phytoplankton. Why do N2 fixers persist in the the oligotrophic North Atlantic where the ratio of nutrients supplied to the surface is elevated in N relative to the average R (16:1)? Combining resource competition theory and a global coupled ecosystem-circulation model we find a novel mechanism that can expand the ecological niche of N2 fixers also to regions where the nutrient supply is high in N relative to R, offering a new perspective on the environmental controls of marine N2-fixers.

  7. North polar region of Mars: imaging results from viking 2.

    PubMed

    Cutts, J A; Blasius, K R; Briggs, G A; Carr, M H; Greeley, R; Masursky, H

    1976-12-11

    During October 1976, the Viking 2 orbiter acquired approximately 700 high-resolution images of the north polar region of Mars. These images confirm the existence at the north pole of extensive layered deposits largely covered over with deposits of perennial ice. An unconformity within the layered deposits suggests a complex history of climate change during their time of deposition. A pole-girdling accumulation of dunes composed of very dark materials is revealed for the first time by the Viking cameras. The entire region is devoid of fresh impact craters. Rapid rates of erosion or deposition are implied. A scenario for polar geological evolution, involving two types of climate change, is proposed.

  8. Brucella abortus surveillance of cattle in Fiji, Papua New Guinea, Vanuatu, the Solomon Islands and a case for active disease surveillance as a training tool.

    PubMed

    Tukana, Andrew; Hedlefs, Robert; Gummow, Bruce

    2016-10-01

    There have been no surveys of the cattle population for brucellosis in the Pacific Island Countries and Territories (PICTs) for more than 15 years. This study used disease surveillance as a capacity building training tool and to examine some of the constraints that impede surveillance in PICTs. The study also developed and implemented a series of surveys for detecting antibodies to B. abortus in cattle in Fiji, Papua New Guinea, Vanuatu and the Solomon Islands contributing to OIE requirements. The findings indicated lack of funds, lack of technical capacity, shortage of veterinarians, high turnover of in-country officials and lack of awareness on the impacts of animal diseases on public health that were constraining active disease surveillance. During the development and implementation of the surveys, constraints highlighted were outdated census data on farm numbers and cattle population, lack of funds for mobilisation of officials to carry out the surveys, lack of equipment for collecting and processing samples, lack of staff knowledge on blood sampling, geographical difficulties and security in accessing farms. Some of the reasons why these were constraints are discussed with likely solutions presented. The detection surveys had the objectives of building capacity for the country officials and demonstrating freedom from brucellosis in cattle for PNG, Vanuatu and the Solomon Islands. PNG, Vanuatu and the Solomon Islands all demonstrated freedom from bovine brucellosis in the areas surveyed using the indirect ELISA test. Fiji had an outbreak of brucellosis, and the objective was to determine its distribution and prevalence on untested farms. The Muaniweni district surveyed during the training had a 95 % confidence interval for true prevalence between 1.66 and 5.45 %. The study showed that active disease surveillance could be used as a tool for training officials thus, improves surveillance capacity in resource poor countries.

  9. The Prevalence of Gap Junction Protein Beta 2 (GJB2) Mutations in Non Syndromic Sensorineural Hearing Loss in Çukurova Region.

    PubMed

    Bozdoğan, Sevcan Tuğ; Kuran, Gökhan; Yüregir, Özge Özalp; Aslan, Hüseyin; Haytoğlu, Süheyl; Ayaz, Akif; Arıkan, Osman Kürşat

    2015-08-01

    To date, studies in all populations showed that mutations in the gene of Gap junction protein beta 2 (GJB2) play an important role in non-syndromic autosomal recessive congenital hearing loss. The aim of this study was to evaluate GJB2 gene of patients with hearing loss in our region using deoxyribonucleic acid (DNA) sequencing method and to demonstrate region-specific mutation and polymorphism distribution. Patients who had bilateral severe sensorineural non-syndromic hearing loss identified by audiologic evaluation were included. Peripheral blood samples were collected and the GJB2 gene exon1 and exon 2 regions were amplified by polymerase chain reaction (PCR). Obtained PCR products were sequenced by the DNA sequence analysis method (SeqFinder Sequencing System; ABI 3130; Foster City, CA, USA) and analyzed using the SeqScape software. Of the 77 patients, 16 had homozygous or heterozygous mutation. The mutation of 35delG, which is known as the most frequent mutation of GJB2 gene, was also the most frequently seen mutation at a ratio of 5.5% in patients with hearing loss in our region; this was followed by the V27I mutation. As this is the first study conducted by sequence analysis in our region, it was worth to be presented in terms of showing the distribution of mutation.

  10. Aura OMI observations of regional SO2 and NO2 pollution changes from 2005 to 2015

    NASA Technical Reports Server (NTRS)

    Krotkov, Nickolay A.; McLinden, Chris A; Li, Can; Lamsal, Lok N.; Celarier, Edward A.; Marchenko, Sergey V.; Swartz, William H.; Bucsela, Eric J.; Joiner, Joanna; Duncan, Bryan N.; hide

    2016-01-01

    The Ozone Monitoring Instrument (OMI) onboard NASA's Aura satellite has been providing global observations of the ozone layer and key atmospheric pollutant gases, such as nitrogen dioxide (NO2/ and sulfur dioxide (SO2/, since October 2004. The data products from the same instrument provide consistent spatial and temporal coverage and permit the study of anthropogenic and natural emissions on local-to-global scales. In this paper, we examine changes in SO2 and NO2 over some of the world's most polluted industrialized regions during the first decade of OMI observations. In terms of regional pollution changes, we see both upward and downward trends, sometimes in opposite directions for NO2 and SO2, for different study areas. The trends are, for the most part, associated with economic and/or technological changes in energy use, as well as regional regulatory policies. Over the eastern US, both NO2 and SO2 levels decreased dramatically from 2005 to 2015, by more than 40 and 80 %, respectively, as a result of both technological improvements and stricter regulations of emissions. OMI confirmed large reductions in SO2 over eastern Europe's largest coal-fired power plants after installation of flue gas desulfurization devices. The North China Plain has the world's most severe SO2 pollution, but a decreasing trend has been observed since 2011, with about a 50% reduction in 2012- 2015, due to an economic slowdown and government efforts to restrain emissions from the power and industrial sectors. In contrast, India's SO2 and NO2 levels from coal power plants and smelters are growing at a fast pace, increasing by more than 100 and 50 %, respectively, from 2005 to 2015. Several SO2 hot spots observed over the Persian Gulf are probably related to oil and gas operations and indicate a possible underestimation of emissions from these sources in bottom-up emission inventories. Overall, OMI observations have proved valuable in documenting rapid changes in air quality over different

  11. Aura OMI Observations of Regional SO2 and NO2 Pollution Changes from 2005 to 2015

    NASA Technical Reports Server (NTRS)

    Krotkov, Nickolay A.; McLinden, Chris A.; Li, Can; Lamsal, Lok N.; Celarier, Edward A.; Marchenko, Sergey V.; Swartz, William H.; Bucsela, Eric J.; Joiner, Joanna; Duncan, Bryan N.; hide

    2016-01-01

    The Ozone Monitoring Instrument (OMI) onboard NASA's Aura satellite has been providing global observations of the ozone layer and key atmospheric pollutant gases, such as nitrogen dioxide (NO2) and sulfur dioxide (SO2), since October 2004. The data products from the same instrument provide consistent spatial and temporal coverage and permit the study of anthropogenic and natural emissions on local-to-global scales. In this paper, we examine changes in SO2 and NO2 over some of the world's most polluted industrialized regions during the first decade of OMI observations. In terms of regional pollution changes, we see both upward and downward trends, sometimes in opposite directions for NO2 and SO2, for different study areas. The trends are, for the most part, associated with economic and/or technological changes in energy use, as well as regional regulatory policies. Over the eastern US, both NO2 and SO2 levels decreased dramatically from 2005 to 2015, by more than 40 and 80 percent, respectively, as a result of both technological improvements and stricter regulations of emissions. OMI confirmed large reductions in SO2 over eastern Europe's largest coal-fired power plants after installation of flue gas desulfurization devices. The North China Plain has the world's most severe SO2 pollution, but a decreasing trend has been observed since 2011, with about a 50 percent reduction in 2012-2015, due to an economic slowdown and government efforts to restrain emissions from the power and industrial sectors. In contrast, India's SO2 and NO2 levels from coal power plants and smelters are growing at a fast pace, increasing by more than 100 and 50 percent, respectively, from 2005 to 2015. Several SO2 hot spots observed over the Persian Gulf are probably related to oil and gas operations and indicate a possible underestimation of emissions from these sources in bottom-up emission inventories. Overall, OMI observations have proved valuable in documenting rapid changes in air

  12. Aura OMI observations of regional SO2 and NO2 pollution changes from 2005 to 2014

    NASA Astrophysics Data System (ADS)

    Krotkov, N. A.; McLinden, C. A.; Li, C.; Lamsal, L. N.; Celarier, E. A.; Marchenko, S. V.; Swartz, W. H.; Bucsela, E. J.; Joiner, J.; Duncan, B. N.; Boersma, K. F.; Veefkind, J. P.; Levelt, P. F.; Fioletov, V. E.; Dickerson, R. R.; He, H.; Lu, Z.; Streets, D. G.

    2015-10-01

    The Ozone Monitoring Instrument (OMI) onboard NASA's Aura satellite has been providing global observations of the ozone layer and key atmospheric pollutant gases, such as nitrogen dioxide (NO2) and sulfur dioxide (SO2), since October 2004. The data products from the same instrument provide consistent spatial and temporal coverage and permit the study of anthropogenic and natural emissions on local-to-global scales. In this paper we examine changes in SO2 and NO2 over some of the world's most polluted industrialized regions during the first decade of OMI observations. In terms of regional pollution changes, we see both upward and downward trends, sometimes in opposite directions for NO2 and SO2, for the different study areas. The trends are, for the most part, associated with economic and/or technological changes in energy use, as well as regional regulatory policies. Over the eastern US, both NO2 and SO2 levels decreased dramatically from 2005 to 2014, by more than 40 and 80 %, respectively, as a result of both technological improvements and stricter regulations of emissions. OMI confirmed large reductions in SO2 over eastern Europe's largest coal power plants after installation of flue gas desulfurization devices. The North China Plain has the world's most severe SO2 pollution, but a decreasing trend has been observed since 2011, with about a 50 % reduction in 2012-2014, due to an economic slowdown and government efforts to restrain emissions from the power and industrial sectors. In contrast, India's SO2 and NO2 levels from coal power plants and smelters are growing at a fast pace, increasing by more than 100 and 50 %, respectively, from 2005 to 2014. Several SO2 hot spots observed over the Persian Gulf are probably related to oil and gas operations and indicate a possible underestimation of emissions from these sources in bottom-up emission inventories. Overall, OMI observations have proved to be very valuable in documenting rapid changes in air quality over

  13. Aura OMI observations of regional SO2 and NO2 pollution changes from 2005 to 2015

    NASA Astrophysics Data System (ADS)

    Krotkov, Nickolay A.; McLinden, Chris A.; Li, Can; Lamsal, Lok N.; Celarier, Edward A.; Marchenko, Sergey V.; Swartz, William H.; Bucsela, Eric J.; Joiner, Joanna; Duncan, Bryan N.; Folkert Boersma, K.; Pepijn Veefkind, J.; Levelt, Pieternel F.; Fioletov, Vitali E.; Dickerson, Russell R.; He, Hao; Lu, Zifeng; Streets, David G.

    2016-04-01

    The Ozone Monitoring Instrument (OMI) onboard NASA's Aura satellite has been providing global observations of the ozone layer and key atmospheric pollutant gases, such as nitrogen dioxide (NO2) and sulfur dioxide (SO2), since October 2004. The data products from the same instrument provide consistent spatial and temporal coverage and permit the study of anthropogenic and natural emissions on local-to-global scales. In this paper, we examine changes in SO2 and NO2 over some of the world's most polluted industrialized regions during the first decade of OMI observations. In terms of regional pollution changes, we see both upward and downward trends, sometimes in opposite directions for NO2 and SO2, for different study areas. The trends are, for the most part, associated with economic and/or technological changes in energy use, as well as regional regulatory policies. Over the eastern US, both NO2 and SO2 levels decreased dramatically from 2005 to 2015, by more than 40 and 80 %, respectively, as a result of both technological improvements and stricter regulations of emissions. OMI confirmed large reductions in SO2 over eastern Europe's largest coal-fired power plants after installation of flue gas desulfurization devices. The North China Plain has the world's most severe SO2 pollution, but a decreasing trend has been observed since 2011, with about a 50 % reduction in 2012-2015, due to an economic slowdown and government efforts to restrain emissions from the power and industrial sectors. In contrast, India's SO2 and NO2 levels from coal power plants and smelters are growing at a fast pace, increasing by more than 100 and 50 %, respectively, from 2005 to 2015. Several SO2 hot spots observed over the Persian Gulf are probably related to oil and gas operations and indicate a possible underestimation of emissions from these sources in bottom-up emission inventories. Overall, OMI observations have proved valuable in documenting rapid changes in air quality over different

  14. 76 FR 33658 - Airworthiness Directives; Bombardier, Inc. Model CL-600-2B19 (Regional Jet Series 100 & 440...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-06-09

    ... Airworthiness Directives; Bombardier, Inc. Model CL-600-2B19 (Regional Jet Series 100 & 440); Model CL-600-2C10 (Regional Jet Series 700, 701, & 702); Model CL-600-2D15 (Regional Jet Series 705); and Model CL-600-2D24 (Regional Jet Series 900) Airplanes AGENCY: Federal Aviation Administration (FAA), DOT. ACTION: Notice of...

  15. 75 FR 47249 - Airworthiness Directives; Bombardier, Inc. Model CL-600-2B19 (Regional Jet Series 100 & 440...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-08-05

    ... DEPARTMENT OF TRANSPORTATION Federal Aviation Administration 14 CFR Part 39 [Docket No. FAA-2010.... Model CL-600-2B19 (Regional Jet Series 100 & 440) Airplanes, CL-600-2C10 (Regional Jet Series 700, 701, & 702) Airplanes, CL-600-2D15 (Regional Jet Series 705) Airplanes, and CL-600-2D24 (Regional Jet Series...

  16. Prevalence of visual impairment in El Salvador: inequalities in educational level and occupational status.

    PubMed

    Rius, Anna; Guisasola, Laura; Sabidó, Meritxell; Leasher, Janet L; Moriña, David; Villalobos, Astrid; Lansingh, Van C; Mujica, Oscar J; Rivera-Handal, José Eduardo; Silva, Juan Casrlos

    2014-11-01

    To examine the prevalence of blindness, visual impairment, and related eye diseases and conditions among adults in El Salvador, and to explore socioeconomic inequalities in their prevalence by education level and occupational status, stratified by sex. Based upon the Rapid Assessment of Avoidable Blindness (RAAB) methodology, this nationwide sample comprised 3 800 participants (3 399 examined) ≥ 50 years old from 76 randomly selected clusters of 50 persons each. The prevalence of blindness, visual impairment and related eye diseases and conditions, including uncorrected refractive error (URE), was calculated for categories of education level and occupational status. Multiple logistic regression models were fitted to calculate odds ratios (ORs) and 95% confidence intervals (CIs) and stratified by sex. Age-adjusted prevalence was 2.4% (95% CI: 2.2-2.6) for blindness (men: 2.8% (95% CI: 2.5-3.1); women: 2.2% (95% CI: 1.9-2.5)) and 11.8% (95% CI: 11.6-12.0) for moderate visual impairment (men: 10.8% (95% CI: 10.5-11.1); women: 12.6% (95% CI: 12.4-12.8)). The proportion of visual impairment due to cataract was 43.8% in men and 33.5% in women. Inverse gradients of socioeconomic inequalities were observed in the prevalence of visual impairment. For example, the age-adjusted OR (AOR) was 3.4 (95% CI: 2.0-6.4) for visual impairment and 4.3 (95% CI: 2.1-10.4) for related URE in illiterate women compared to those with secondary education, and 1.9 (95% CI: 1.1-3.1) in cataract in unemployed men. Blindness and visual impairment prevalence is high in the El Salvador adult population. The main associated conditions are cataract and URE, two treatable conditions. As socioeconomic and gender inequalities in ocular health may herald discrimination and important barriers to accessing affordable, good-quality, and timely health care services, prioritization of public eye health care and disability policies should be put in place, particularly among women, the unemployed, and

  17. SOLIS IV. Hydrocarbons in the OMC-2 FIR4 Region, a Probe of Energetic Particle Irradiation of the Region

    NASA Astrophysics Data System (ADS)

    Favre, C.; Ceccarelli, C.; López-Sepulcre, A.; Fontani, F.; Neri, R.; Manigand, S.; Kama, M.; Caselli, P.; Jaber Al-Edhari, A.; Kahane, C.; Alves, F.; Balucani, N.; Bianchi, E.; Caux, E.; Codella, C.; Dulieu, F.; Pineda, J. E.; Sims, I. R.; Theulé, P.

    2018-06-01

    We report new interferometric images of cyclopropenylidene, c-C3H2, toward the young protocluster OMC-2 FIR 4. The observations were performed at 82 and 85 GHz with the NOrthern Extended Millimeter Array (NOEMA) as part of the project Seeds Of Life In Space (SOLIS). In addition, IRAM-30 m data observations were used to investigate the physical structure of OMC-2 FIR 4. We find that the c-C3H2 gas emits from the same region where previous SOLIS observations showed bright HC5N emission. From a non-LTE analysis of the IRAM-30 m data, the c-C3H2 gas has an average temperature of ∼40 K, a H2 density of ∼3 × 105 cm‑3, and a c-C3H2 abundance relative to H2 of (7 ± 1) × 10‑12. In addition, the NOEMA observations provide no sign of significant c-C3H2 excitation temperature gradients across the region (about 3–4 beams), with T ex in the range 8 ± 3 up to 16 ± 7 K. We thus infer that our observations are inconsistent with a physical interaction of the OMC-2 FIR 4 envelope with the outflow arising from OMC-2 FIR 3, as claimed by previous studies. The comparison of the measured c-C3H2 abundance with the predictions from an astrochemical PDR model indicates that OMC-2 FIR 4 is irradiated by an FUV field ∼1000 times larger than the interstellar one, and by a flux of ionizing particles ∼4000 times larger than the canonical value of 1 × 10‑17 s‑1 from the Galaxy cosmic rays, which is consistent with our previous HC5N observations. This provides an important and independent confirmation of other studies that one, or more, source inside the OMC-2 FIR 4 region emits energetic (≥10 MeV) particles. Based on observations carried out under project number L15AA with the IRAM NOEMA Interferometer. IRAM is supported by INSU/CNRS (France), MPG (Germany) and IGN (Spain).

  18. The first CO+ image: I. Probing the HI/H2 layer around the ultracompact HII region Mon R2

    PubMed Central

    Treviño-Morales, S. P.; Fuente, A.; Sánchez-Monge, Á.; Pilleri, P.; Goicoechea, J. R.; Ossenkopf-Okada, V.; Roueff, E.; Rizzo, J. R.; Gerin, M.; Berné, O.; Cernicharo, J.; Gónzalez-García, M.; Kramer, C.; García-Burillo, S.; Pety, J.

    2016-01-01

    The CO+ reactive ion is thought to be a tracer of the boundary between a HII region and the hot molecular gas. In this study, we present the spatial distribution of the CO+ rotational emission toward the Mon R2 star-forming region. The CO+ emission presents a clumpy ring-like morphology, arising from a narrow dense layer around the HII region. We compare the CO+ distribution with other species present in photon-dominated regions (PDR), such as [CII] 158 µm, H2 S(3) rotational line at 9.3 µm, polycyclic aromatic hydrocarbons (PAHs) and HCO+. We find that the CO+ emission is spatially coincident with the PAHs and [CII] emission. This confirms that the CO+ emission arises from a narrow dense layer of the HI/H2 interface. We have determined the CO+ fractional abundance, relative to C+ toward three positions. The abundances range from 0.1 to 1.9 ×10−10 and are in good agreement with previous chemical model, which predicts that the production of CO+ in PDRs only occurs in dense regions with high UV fields. The CO+ linewidth is larger than those found in molecular gas tracers, and their central velocity are blue-shifted with respect to the molecular gas velocity. We interpret this as a hint that the CO+ is probing photo-evaporating clump surfaces. PMID:27721515

  19. Mapping CO2 emission in highly urbanized region using standardized microbial respiration approach

    NASA Astrophysics Data System (ADS)

    Vasenev, V. I.; Stoorvogel, J. J.; Ananyeva, N. D.

    2012-12-01

    Urbanization is a major recent land-use change pathway. Land conversion to urban has a tremendous and still unclear effect on soil cover and functions. Urban soil can act as a carbon source, although its potential for CO2 emission is also very high. The main challenge in analysis and mapping soil organic carbon (SOC) in urban environment is its high spatial heterogeneity and temporal dynamics. The urban environment provides a number of specific features and processes that influence soil formation and functioning and results in a unique spatial variability of carbon stocks and fluxes at short distance. Soil sealing, functional zoning, settlement age and size are the predominant factors, distinguishing heterogeneity of urban soil carbon. The combination of these factors creates a great amount of contrast clusters with abrupt borders, which is very difficult to consider in regional assessment and mapping of SOC stocks and soil CO2 emission. Most of the existing approaches to measure CO2 emission in field conditions (eddy-covariance, soil chambers) are very sensitive to soil moisture and temperature conditions. They require long-term sampling set during the season in order to obtain relevant results. This makes them inapplicable for the analysis of CO2 emission spatial variability at the regional scale. Soil respiration (SR) measurement in standardized lab conditions enables to overcome this difficulty. SR is predominant outgoing carbon flux, including autotrophic respiration of plant roots and heterotrophic respiration of soil microorganisms. Microbiota is responsible for 50-80% of total soil carbon outflow. Microbial respiration (MR) approach provides an integral CO2 emission results, characterizing microbe CO2 production in optimal conditions and thus independent from initial difference in soil temperature and moisture. The current study aimed to combine digital soil mapping (DSM) techniques with standardized microbial respiration approach in order to analyse and

  20. A Chandra X-ray Mosaic of the Onsala 2 Star-Forming Region

    NASA Astrophysics Data System (ADS)

    Skinner, Steve L.; Sokal, Kimberly; Guedel, Manuel

    2018-01-01

    Multiple lines of evidence for active high-mass star-formation in the Onsala 2 (ON2) complex in Cygnus include masers, compact HII (cHII) regions, and massive outflows. ON2 is thought to be physically associated with the young stellar cluster Berkeley 87 which contains several optically-identified OB stars and the rare oxygen-type (WO) Wolf-Rayet star WR 142. WO stars are undergoing advanced nuclear core burning as they approach the end of their lives as supernovae, and only a few are known in the Galaxy. We present results of a sensitive 70 ks Chandra ACIS-I observation of the northern half of ON2 obtained in 2016. This new observation, when combined with our previous 70 ks ACIS-I observation of the southern half in 2009, provides a complete X-ray mosaic of ON2 at arcsecond spatial resolution and reveals several hundred X-ray sources. We will summarize key results emerging from our ongoing analysis including the detection of an embedded population of young stars revealed as a tight grouping of X-ray sources surrounding the cHII region G75.77+0.34, possible diffuse X-ray emission (or unresolved faint point sources) near the cHII region G75.84+0.40, and confirmation of hard heavily-absorbed X-ray emission from WR 142 that was seen in the previous 2009 Chandra observation.

  1. DNA Methylation Analysis of HTR2A Regulatory Region in Leukocytes of Autistic Subjects.

    PubMed

    Hranilovic, Dubravka; Blazevic, Sofia; Stefulj, Jasminka; Zill, Peter

    2016-02-01

    Disturbed brain and peripheral serotonin homeostasis is often found in subjects with autism spectrum disorder (ASD). The role of the serotonin receptor 2A (HTR2A) in the regulation of central and peripheral serotonin homeostasis, as well as its altered expression in autistic subjects, have implicated the HTR2A gene as a major candidate for the serotonin disturbance seen in autism. Several studies, yielding so far inconclusive results, have attempted to associate autism with a functional SNP -1438 G/A (rs6311) in the HTR2A promoter region, while possible contribution of epigenetic mechanisms, such as DNA methylation, to HTR2A dysregulation in autism has not yet been investigated. In this study, we compared the mean DNA methylation within the regulatory region of the HTR2A gene between autistic and control subjects. DNA methylation was analysed in peripheral blood leukocytes using bisulfite conversion and sequencing of the HTR2A region containing rs6311 polymorphism. Autistic subjects of rs6311 AG genotype displayed higher mean methylation levels within the analysed region than the corresponding controls (P < 0.05), while there was no statistically significant difference for AA and GG carriers. Our study provides preliminary evidence for increased HTR2A promoter methylation in leukocytes of a portion of adult autistic subjects, indicating that epigenetic mechanisms might contribute to HTR2A dysregulation observed in individuals with ASD. © 2015 International Society for Autism Research, Wiley Periodicals, Inc.

  2. Production of infectious chimeric hepatitis C virus genotype 2b harboring minimal regions of JFH-1.

    PubMed

    Murayama, Asako; Kato, Takanobu; Akazawa, Daisuke; Sugiyama, Nao; Date, Tomoko; Masaki, Takahiro; Nakamoto, Shingo; Tanaka, Yasuhito; Mizokami, Masashi; Yokosuka, Osamu; Nomoto, Akio; Wakita, Takaji

    2012-02-01

    To establish a cell culture system for chimeric hepatitis C virus (HCV) genotype 2b, we prepared a chimeric construct harboring the 5' untranslated region (UTR) to the E2 region of the MA strain (genotype 2b) and the region of p7 to the 3' UTR of the JFH-1 strain (genotype 2a). This chimeric RNA (MA/JFH-1.1) replicated and produced infectious virus in Huh7.5.1 cells. Replacement of the 5' UTR of this chimera with that from JFH-1 (MA/JFH-1.2) enhanced virus production, but infectivity remained low. In a long-term follow-up study, we identified a cell culture-adaptive mutation in the core region (R167G) and found that it enhanced virus assembly. We previously reported that the NS3 helicase (N3H) and the region of NS5B to 3' X (N5BX) of JFH-1 enabled replication of the J6CF strain (genotype 2a), which could not replicate in cells. To reduce JFH-1 content in MA/JFH-1.2, we produced a chimeric viral genome for MA harboring the N3H and N5BX regions of JFH-1, combined with a JFH-1 5' UTR replacement and the R167G mutation (MA/N3H+N5BX-JFH1/R167G). This chimeric RNA replicated efficiently, but virus production was low. After the introduction of four additional cell culture-adaptive mutations, MA/N3H+N5BX-JFH1/5am produced infectious virus efficiently. Using this chimeric virus harboring minimal regions of JFH-1, we analyzed interferon sensitivity and found that this chimeric virus was more sensitive to interferon than JFH-1 and another chimeric virus containing more regions from JFH-1 (MA/JFH-1.2/R167G). In conclusion, we established an HCV genotype 2b cell culture system using a chimeric genome harboring minimal regions of JFH-1. This cell culture system may be useful for characterizing genotype 2b viruses and developing antiviral strategies.

  3. Apatite and Chitin Amendments Promote Microbial Activity and Augment Metal Removal in Marine Sediments

    DTIC Science & Technology

    2013-07-01

    abundance of Deltaproteobacteria and decrease of Actinobacteria , Alphaproteobacteria, and Nitrospirae. Many of these groups of bacteria have been shown to...shifts were observed from clone library analyses (Table 2 and Fig- ure 4). Actinobacteria , Alphaproteobacteria, Gam- maproteobacteria...abundance (%) Phylum FUS FCS FACS Impact Acidobacteria - - 1.9 Induced Actinobacteria 20.8 8.6 3.8 Decreased Alphaproteobacteria 22.9 14.3 13.2 Decreased

  4. Deep optical survey of the stellar content of Sh2-311 region

    NASA Astrophysics Data System (ADS)

    Yadav, Ram Kesh; Pandey, A. K.; Sharma, Saurabh; Jose, J.; Ogura, K.; Kobayashi, N.; Samal, M. R.; Eswaraiah, C.; Chandola, H. C.

    2015-01-01

    The stellar content in and around Sh2-311 region have been studied using the deep optical observations as well as near-infrared (NIR) data from 2MASS. The region contains three clusters, viz. NGC 2467, Haffner 18 and Haffner 19. We have made an attempt to distinguish the stellar content of these individual regions as well as to re-determine their fundamental parameters such as distance, reddening, age, onto the basis of a new and more extended optical and infrared photometric data set. NGC 2467 and Haffner 19 are found to be located in the Perseus arm at the distances of 5.0 ± 0.4 kpc and 5.7 ± 0.4 kpc, respectively, whereas Haffner 18 is located at the distance of 11.2 ± 1.0 kpc. The clusters NGC 2467 and Haffner 19 might have formed from the same molecular cloud, whereas the cluster Haffner 18 is located in the outer galactic arm, i.e. the Norma-Cygnus arm. We identify 8 class II young stellar objects (YSOs) using the NIR (J-H)/(H-K) two colour diagram. We have estimated the age and mass of the YSOs identified in the present work and those by Snider et al. (2009) using the V/(V-I) colour-magnitude diagram. The estimated ages and mass range of the majority of the YSOs are ≲1 Myr and ∼0.4-3.5 M⊙, respectively, indicating that these sources could be T-Tauri stars or their siblings. Spatial distribution of the YSOs shows that some of the YSOs are distributed around the HII region Sh2-311, suggesting a triggered star formation at its periphery.

  5. 78 FR 44909 - Regional Reliability Standard BAL-002-WECC-2-Contingency Reserve

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-07-25

    ...] Regional Reliability Standard BAL-002-WECC-2--Contingency Reserve AGENCY: Federal Energy Regulatory...-002-WECC-2 (Contingency Reserve). The North American Electric Reliability Corporation (NERC) and... contingency reserve required to ensure reliability under normal and abnormal conditions. The Commission also...

  6. Regional N2O fluxes in Amazonia derived from aircraft vertical profiles

    NASA Astrophysics Data System (ADS)

    D'Amelio, M. T. S.; Gatti, L. V.; Miller, J. B.; Tans, P.

    2009-11-01

    Nitrous oxide (N2O) is the third most important anthropogenic greenhouse gas. Globally, the main sources of N2O are nitrification and denitrification in soils. About two thirds of the soil emissions occur in the tropics and approximately 20% originate in wet rainforest ecosystems, like the Amazon forest. The work presented here involves aircraft vertical profiles of N2O from the surface to 4 km over two sites in the Eastern and Central Amazon: Tapajós National Forest (SAN) and Cuieiras Biologic Reserve (MAN), and the estimation of N2O fluxes for regions upwind of these sites. To our knowledge, these regional scale N2O measurements in Amazonia are unique and represent a new approach to looking regional scale emissions. The fluxes upwind of MAN exhibited little seasonality, and the annual mean was 2.1±1.0 mg N2O m-2 day-1, higher than that for fluxes upwind of SAN, which averaged 1.5±1.6 mg N2O m-2 day-1. The higher rainfall around the MAN site could explain the higher N2O emissions, as a result of increased soil moisture accelerating microbial nitrification and denitrification processes. For fluxes from the coast to SAN seasonality is present for all years, with high fluxes in the months of March through May, and in November through December. The first peak of N2O flux is strongly associated with the wet season. The second peak of high N2O flux recorded at SAN occurs during the dry season and can not be easily explained. However, about half of the dry season profiles exhibit significant correlations with CO, indicating a larger than expected source of N2O from biomass burning. The average CO:N2O ratio for all profiles sampled during the dry season is 94±77 mol CO:mol N2O and suggests a larger biomass burning contribution to the global N2O budget than previously reported.

  7. Regional N2O fluxes in Amazonia derived from aircraft vertical profiles

    NASA Astrophysics Data System (ADS)

    D'Amelio, M. T. S.; Gatti, L. V.; Miller, J. B.; Tans, P.

    2009-08-01

    Nitrous oxide (N2O) is the third most important anthropogenic greenhouse gas. Globally, the main sources of N2O are nitrification and denitrification in soils. About two thirds of the soil emissions occur in the tropics and approximately 20% originate in wet rainforest ecosystems, like the Amazon forest. The work presented here involves aircraft vertical profiles of N2O from the surface to 4 km over two sites in the Eastern and Central Amazon: Tapajós National Forest (SAN) and Cuieiras Biologic Reserve (MAN), and the estimation of N2O fluxes for regions upwind of these sites. To our knowledge, these regional scale N2O measurements in Amazonia are unique and represent a new approach to looking regional scale emissions. The fluxes upwind of MAN exhibited little seasonality, and the annual mean was 2.1±1.0 mg N2O m-2 day-1, higher than that for fluxes upwind of SAN, which averaged 1.5±1.6 mg N2O m-2 day-1. The higher rainfall around the MAN site could explain the higher N2O emissions. For fluxes from the coast to SAN seasonality is present for all years, with high fluxes in the months of March through May, and in November through December. The first peak of N2O flux is strongly associated with the wet season. The second peak of high N2O flux recorded at SAN occurs during the dry season and can not be easily explained. However, about half of the dry season profiles exhibit significant correlations with CO, indicating a larger than expected source of N2O from biomass burning. The average CO:N2O ratio for all profiles sampled during the dry season is 94±77 mol CO:mol N2O and suggests a larger biomass burning contribution to the global N2O budget than previously reported.

  8. Serological trail of Brucella infection in an urban slum population in Brazil

    PubMed Central

    Angel, Martha Olivera; Ristow, Paula; Ko, Albert I.; Di-Lorenzo, Cecilia

    2013-01-01

    Introduction Brucellosis is a re-emerging zoonosis with new cases reported each year in many Latin American countries, but it is mostly under-recognized. This study presents a serological investigation of infection with Brucella abortus and Brucella canis in a poor urban community in the city of Salvador, Brazil. Methodology Human sera (n = 180) were randomly selected from 3,171 samples taken from healthy individuals during 2003-2004 and tested with C-ELISA for B. abortus and I-ELISA for B. canis. Results Thirteen percent (24/180) of the individuals were positive for B. abortus and 4.6 % (8/174) were positive for B. canis. Among the variables studied only age (older than 45 years) appeared to be a risk factor for the detection of Brucella antibodies. Conclusion These results indicate the presence of Brucella infection in this settlement and highlight the need to understand the epidemiology of infection under these circumstances to establish the necessary measures for surveillance and control. PMID:23000868

  9. Environmentally Sensitive Area (ESA) Grant Condition Waiver Program in Region 2

    EPA Pesticide Factsheets

    Starting in October 1979, EPA Region 2 applied a special condition to many wastewater facilities’ construction grants (see Federal Register, Vol. 43, No. 188, September 27, 1978) to protect Environmentally Sensitive Areas(EASs) from induced development.

  10. Hurricane Ivan’s Impact Along the Northern Gulf of Mexico. Volume 86, Number 48

    DTIC Science & Technology

    2005-11-29

    M e 17.91-meter-high waves at the same buoy East of the Chandeleur Island arc along southeastern Fig. 1. (a) MODIS nighttime sea surface temperature...MODIS image from 16 September (Fig- ure 2a) revealed a large mass of sediment flowing southward along the east side of the Chandeleur Islands and

  11. Io's Thermal Regions and Non-SO2 Spectral Features

    NASA Technical Reports Server (NTRS)

    Smythe, W. D.; Soderblom, L. A.; Lopes, R. M. C.

    2003-01-01

    Several absorptions have been identified in the Galileo NIMS spectra of Io that are not related to SO2. [1,2]. These absorptions have band centers at 2.97, 3.15, 3.85, and 3.91 microns. There are also broad absorptions in the regions 1-1.3 and 3- 3.4 microns. Patterning noise in wavelength registration, arising from the pushbroom imaging and grating motion of the NIMS instrument have previously inhibited reliable mapping of weak absorptions. Recent improvements in techniques to remove the coherent pattern noise from the NIMS dataset have been made by Soderblom. This greatly improves the signal to noise ratio and enables mapping of weak spectral signatures such as the 3.15 micron absorption on Io.

  12. Coast to Coast Support of the E-2C Hawkeye using Distributed TSP

    DTIC Science & Technology

    2008-05-02

    Share history of each subgroup and establish a vision f th f t f thi l f d to e u ure or s new y orme eam 3. Establish team operating principles...visits were made by team t th ld k t b d b ild th b id b t th Th ff t NAVAIR Systems/Software Support Center (NSSC) Slide 18 managemen so ey cou now...Success • High quality products from multiple teams delivered on ti d t d tme an cos o no happen in a vacuum • There is a need for common processes

  13. El medio interestelar en los alrededores de la region HII Sh2-183

    NASA Astrophysics Data System (ADS)

    Cichowolski, S.; Cappa, C. E.; Blanco, A.; Eppens, L.; Ertini, K.; Leiva, M. M.

    2017-10-01

    We present a multiwavelength study of the HII region Sh2-183, located at (,) = (123.3,+3.0) at a distance of 7.0 1.5 kpc from the Sun. Based on the radio continuum data we estimated the amount of ionized gas, the electronic density, and the number of ionizing photons needed to keep the region ionized, which is important since the star/s responsible of the region was/were not detected yet. On the other hand, based on IRAS data we have analyzed the dust temperature and distribution. The Hi line data allowed the detection of a shell-like structure surrounding the ionized gas and the CO data revealed the presence of 6 molecular clouds probably related to Sh2-183, which harbor several young stellar object candidates.

  14. Mapping the stability region of the 3:2 Neptune-Pluto resonance

    NASA Technical Reports Server (NTRS)

    Levison, H. F.; Stern, S. A.

    1993-01-01

    Pluto and Charon are most likely the remnants of a large number of objects that existed in the Uranus-Neptune region at early epochs of the solar system. Numerical integrations have shown that, in general, such objects were ejected from the planetary region on timescales of approximately 10(exp 7) years after Neptune and Uranus reached their current masses. It is thought that the Pluto-Charon system survived to current times without being dynamically removed in this way because it is trapped in a set of secular and mean motion resonances with Neptune. The best-known Pluto-Neptune orbit coupling is the 3:2 mean motion resonance discovered almost 30 years ago by C. Cohen and E. Hubbard. These workers showed that the resonance angle, delta is equivalent to 3(lambda(sub P)) - 2(lambda(sub N)) - omega-bar(sub P) where omega-bar(sub P) is the longitude of perihelion of the Pluto-Charon system, and lambda(sub N) and lambda(sub P) are the mean longitude of Neptune and Pluto-Charon respectively, librates about 180 deg with an amplitude, A(sub delta), of 76 deg. A numerical simulation project to map out the stability region of the 3:2 resonance is reported. The results of these simulations are important to understanding whether Pluto's long-term heliocentric stability requires only the 3:2 resonance, or whether it instead requires one or more of the other Pluto-Neptune resonances. Our study also has another important application. By investigating stability timescales as a function of orbital elements, we gain insight into the fraction of orbital phase space which the stable 3:2 resonance occupies. This fraction is directly related to the probability that the Pluto-Charon system (and possibly other small bodies) could have been captured into this resonance.

  15. Specific inhibition of c-Jun N-terminal kinase delays preterm labour and reduces mortality

    PubMed Central

    Pirianov, Grisha; MacIntyre, David A; Lee, Yun; Waddington, Simon N; Terzidou, Vasso; Mehmet, Huseyin; Bennett, Phillip R

    2015-01-01

    Preterm labour (PTL) is commonly associated with infection and/or inflammation. Lipopolysaccharide (LPS) from different bacteria can be used to independently or mutually activate Jun N-terminal kinase (JNK)/AP1- or NF-κB-driven inflammatory pathways that lead to PTL. Previous studies using Salmonella abortus LPS, which activates both JNK/AP-1 and NF-κB, showed that selective inhibition of NF-κB delays labour and improves pup outcome. Where labour is induced using Escherichia coli LPS (O111), which upregulates JNK/AP-1 but not NF-κB, inhibition of JNK/AP-1 activation also delays labour. In this study, to determine the potential role of JNK as a therapeutic target in PTL, we investigated the specific contribution of JNK signalling to S. Abortus LPS-induced PTL in mice. Intrauterine administration of S. Abortus LPS to pregnant mice resulted in the activation of JNK in the maternal uterus and fetal brain, upregulation of pro-inflammatory proteins COX-2, CXCL1, and CCL2, phosphorylation of cPLA2 in myometrium, and induction of PTL. Specific inhibition of JNK by co-administration of specific D-JNK inhibitory peptide (D-JNKI) delayed LPS-induced preterm delivery and reduced fetal mortality. This is associated with inhibition of myometrial cPLA2 phosphorylation and proinflammatory proteins synthesis. In addition, we report that D-JNKI inhibits the activation of JNK/JNK3 and caspase-3, which are important mediators of neural cell death in the neonatal brain. Our data demonstrate that specific inhibition of TLR4-activated JNK signalling pathways has potential as a therapeutic approach in the management of infection/inflammation-associated PTL and prevention of the associated detrimental effects to the neonatal brain. PMID:26183892

  16. Transition from the Unipolar Region to the Sector Zone: Voyager 2, 2013 and 2014

    NASA Astrophysics Data System (ADS)

    Burlaga, L. F.; Ness, N. F.; Richardson, J. D.

    2017-05-01

    We discuss magnetic field and plasma observations of the heliosheath made by Voyager 2 (V2) during 2013 and 2014 near solar maximum. A transition from a unipolar region to a sector zone was observed in the azimuthal angle λ between ˜2012.45 and 2013.82. The distribution of λ was strongly singly peaked at 270^\\circ in the unipolar region and double peaked in the sector zone. The δ-distribution was strongly peaked in the unipolar region and very broad in the sector zone. The distribution of daily averages of the magnetic field strength B was Gaussian in the unipolar region and lognormal in the sector zone. The correlation function of B was exponential with an e-folding time of ˜5 days in both regions. The distribution of hourly increments of B was a Tsallis distribution with nonextensivity parameter q = 1.7 ± 0.04 in the unipolar region and q = 1.44 ± 0.12 in the sector zone. The CR-B relationship qualitatively describes the 2013 observations, but not the 2014 observations. A 40 km s-1 increase in the bulk speed associated with an increase in B near 2013.5 might have been produced by the merging of streams. A “D sheet” (a broad depression in B containing a current sheet moved past V2 from days 320 to 345, 2013. The R- and N-components of the plasma velocity changed across the current sheet.

  17. Interservice/Industry Training Equipment Conference and Exhibition (3rd) , Orlando, Florida, 30 November-2 December 1981

    DTIC Science & Technology

    1981-01-01

    visual system. Sons of these advantages are "Photogammetry", Harper and Ro, its ability to provide scene-realism, detail, Third Edition, New York...was introduced to the A- la cockpit of the ASPT byl 58 an instructor pilot who gave him a standardized, most difficult turns (45, 60, and 90-degree ten...1 61T 4TOPN *ITO, RA. EL. DJ. ASSY. - UA . E L . ADJ. A llY.1 1 \\a EGUL TORF 0- 30 Psi *LIQUID 0O STORAGE~ PR &URE REG LA O-0!ioPSI 1Pi - YaI~ ~ICH

  18. Domain Engineered Magnetoelectric Thin Films for High Sensitivity Resonant Magnetic Field Sensors

    DTIC Science & Technology

    2012-02-28

    texture E analysis w cated by poo re accounted n measurem 8 sol-gel samp d PZT sol-g as utilized t r fit between in the mo ent spot). les shown i el...nsformer str nted by aero ure. ure 34: Un were grow as varied in D) as show texturing in . D pattern of the films d ucture. Figu sol jet depo ipoled PZT ...the detailed characterization was the development of prediction models for texturing of PZT sol-gel thin films, an understanding of the analytical

  19. 28 CFR 2.24 - Review of panel recommendation by the Regional Commissioner.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 28 Judicial Administration 1 2010-07-01 2010-07-01 false Review of panel recommendation by the Regional Commissioner. 2.24 Section 2.24 Judicial Administration DEPARTMENT OF JUSTICE PAROLE, RELEASE, SUPERVISION AND RECOMMITMENT OF PRISONERS, YOUTH OFFENDERS, AND JUVENILE DELINQUENTS United States Code...

  20. 28 CFR 2.24 - Review of panel recommendation by the Regional Commissioner.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 28 Judicial Administration 1 2011-07-01 2011-07-01 false Review of panel recommendation by the Regional Commissioner. 2.24 Section 2.24 Judicial Administration DEPARTMENT OF JUSTICE PAROLE, RELEASE, SUPERVISION AND RECOMMITMENT OF PRISONERS, YOUTH OFFENDERS, AND JUVENILE DELINQUENTS United States Code...

  1. Observations of Atmospheric Δ14CO2 at the Global and Regional Background Sites in China: Implication for Fossil Fuel CO2 Inputs.

    PubMed

    Niu, Zhenchuan; Zhou, Weijian; Cheng, Peng; Wu, Shugang; Lu, Xuefeng; Xiong, Xiaohu; Du, Hua; Fu, Yunchong

    2016-11-15

    Six months to more than one year of atmospheric Δ 14 CO 2 were measured in 2014-2015 at one global background site in Waliguan (WLG) and four regional background sites at Shangdianzi (SDZ), Lin'an (LAN), Longfengshan (LFS) and Luhuitou (LHT), China. The objectives of the study are to document the Δ 14 CO 2 levels at each site and to trace the variations in fossil fuel CO 2 (CO 2ff ) inputs at regional background sites. Δ 14 CO 2 at WLG varied from 7.1 ± 2.9‰ to 32.0 ± 3.2‰ (average 17.1 ± 6.8‰) in 2015, with high values generally in autumn/summer and low values in winter/spring. During the same period, Δ 14 CO 2 values at the regional background sites were found to be significantly (p < 0.05) lower than those at WLG, indicating different levels of CO 2ff inputs at those sites. CO 2ff concentrations at LAN (12.7 ± 9.6 ppm) and SDZ (11.5 ± 8.2 ppm) were significantly (p < 0.05) higher than those at LHT (4.6 ± 4.3 ppm) in 2015. There were no significant (p > 0.05) seasonal differences in CO 2ff concentrations for the regional sites. Regional sources contributed in part to the CO 2ff inputs at LAN and SDZ, while local sources dominated the trend observed at LHT. These data provide a preliminary understanding of atmospheric Δ 14 CO 2 and CO 2ff inputs for a range of Chinese background sites.

  2. Analysis of uncertainties in GOSAT-inferred regional CO2 fluxes

    NASA Astrophysics Data System (ADS)

    Ishizawa, M.; Shirai, T.; Maksyutov, S. S.; Yoshida, Y.; Morino, I.; Inoue, M.; Nakatsuru, T.; Uchino, O.; Mabuchi, K.

    2016-12-01

    Satellite-based CO2 measurements have potential for improving our understanding global carbon cycle because of more spatiotemporal coverage than those from ground-based observations. Since the Greenhouse gases Observing Satellite (GOSAT) was launched in January 2009, it has been measuring the column-average dry air-mole function of CO2 (XCO2) from the space. To utilize the GOSAT XCO2 for better CO2 flux estimates, several challenges should be overcome. Systematic errors (biases) in XCO2 retrievals are a major factor which leads to large differences among inverted CO2 fluxes. Temporally variable data coverage and density are also taken into account when interpreting the estimated surface fluxes. In this study, we employ an atmospheric inverse model to investigate the impacts of retrievals biases and temporally varying global distribution of GOSAT XCO2 on surface CO2 flux estimates. Inversions are performed for 2009-2013, with several subsets of the 5-year record of GOSAT XCO2 (v2.21) and its bias-corrected XCO2. GOSAT XCO2 data consist of three types: H-gain for vegetated lands, M-gain for bright surfaces (desert areas), and sun-glint for ocean surface. The results show that the global spatial distributions of estimated CO2 fluxes depend on the subset of XCO2 used. M-gain XCO2 results in unrealistically high CO2 emissions in and around the Middle East, including the neighboring ocean regions. On the other hand, M-gain XCO2 causes compensating unrealistic uptakes far beyond M-gain regions in low latitudes, also partially contributing on the summer uptake in Europe. The joint inversions with both surface measurements and GOSAT XCO2 data obtain larger flux gradient between the northern extra-tropics and the tropics than the inversion with surface measurements only for the first 2 years. Recently, these North-South gradients seem to be gradually reducing as the tropics become a weaker source or turn into a sink, while the net emission strength in East Asia is increasing

  3. [Influence of different slope position and profile in Disporopsis pernyi forest land on soil microbial biomass and enzyme activity in southwest Karst mountain of China ].

    PubMed

    Qin, Hua-Jun; He, Bing-Hui; Zhao, Xuan-chi; Li, Yuan; Mao, Wen-tao; Zeng, Qing-ping

    2014-09-01

    Soil microbial biomass and enzyme activity are important parameters to evaluate the quality of the soil environment. The goal of this study was to determine the influence of different slope position and section in Disporopsis pernyi forest land on the soil microbial biomass and enzyme activity in southwest Karst Mountain. In this study, we chose the Dip forest land at Yunfo village Chengdong town Liangping country Chongqing Province as the study object, to analyze the influence of three different slope positions [Up Slope(US), Middle Slope(MS), Below Slope(BS)] and two different sections-upper layer(0-15 cm) and bottom layer(15-30 cm) on the soil microbial biomass carbon (SMBC), soil microbial biomass nitrogen (SMBN), microbial carbon entropy (qMBC), microbial nitrogen entropy (qMBN) , catalase(CAT), alkaline phosphatase (ALK), urease(URE), and invertase(INV). The results showed that the same trend (BS > MS > US) was found for SMBC, SMBN, qMBC, qMBN, CAT and INV of upper soil layer, while a different trend (BS > US > MS) was observed for ALK. In addition, another trend (MS > US > BS) was observed for URE. The same trend (BS > MS >US) was observed for SMBN, qMBN, CAT, ALK, URE and INV in bottom layer, but a different trend (MS > BS > US) was observed for SMBC and qMBC. The SMBC, SMBN, CAT, ALK, URE and INV manifested as upper > bottom with reduction of the section, while qMBC and qMBN showed the opposite trend. Correlation analysis indicated that there were significant (P <0.05) or highly significant (P < 0.01) positive correlations among SMBC in different slope position and section, soil enzyme activity and moisture. According to the two equations of regression analysis, SMBC tended to increase with the increasing CAT and ALK, while decreased with the increasing pH. Then SMBN tended to increase with the increasing URE and INV.

  4. An Undergraduate Research Experience that Integrates Traditional Field Mapping, LiDAR, and 3D Numerical Modeling: Applying Lessons from a Recent Report from the National Academies of Sciences, Engineering, and Medicine in an Intermediate-Level Tectonic Landscapes Course

    NASA Astrophysics Data System (ADS)

    Reinen, L. A.; Brenner, K.

    2017-12-01

    Ongoing efforts to improve undergraduate education in science, technology, engineering, and mathematics (STEM) fields focus on increasing active student participation and decreasing traditional lecture-based teaching. Undergraduate research experiences (UREs), which engage students in the work of STEM professionals, are an example of these efforts. A recent report from the National Academies of Sciences, Engineering and Medicine (Undergraduate Research Experiences for STEM Students: Successes, Challenges, and Opportunities; 2017) provides characteristics of UREs, and indicates that participation in UREs increases student interest and persistence in STEM as well as provides opportunities to broaden student participation in these fields. UREs offer an excellent opportunity to engage students in research using the rapidly evolving technologies used by STEM professionals. In the fall of 2016, students in the Tectonic Landscapes class at Pomona College participated in a course-based URE that combined traditional field mapping methods with analysis of high-resolution topographic data (LiDAR) and 3D numerical modeling to investigate questions of active local faulting. During the first ten weeks students developed skills in: creation of fault maps from both field observations (GPS included) and high-resolution digital elevation models (DEMs), assessment of tectonic activity through analyses of DEMs of hill slope diffusion models and geomorphic indices, and evaluation of fault geometry hypotheses via 3D elastic modeling. Most of these assignments were focused on a single research site. While students primarily used Excel, ArcMap, and Poly3D, no previous knowledge of these was required or assumed. Through this iterative approach, students used increasingly more complex methods as well as gained greater ownership of the research process with time. The course culminated with a 4-week independent research project in which each student investigated a question of their own

  5. 75 FR 12152 - Airworthiness Directives; Bombardier, Inc. Model CL-600-2B19 (Regional Jet Series 100 & 440), CL...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-03-15

    ... DEPARTMENT OF TRANSPORTATION Federal Aviation Administration 14 CFR Part 39 [Docket No. FAA-2010.... Model CL-600-2B19 (Regional Jet Series 100 & 440), CL-600-2C10 (Regional Jet Series 700, 701 & 702), CL-600-2D15 (Regional Jet Series 705), and CL-600-2D24 (Regional Jet Series 900) Airplanes AGENCY...

  6. 41 CFR 101-25.110-2 - Tires obtained through Federal Supply Schedules or regional term contracts.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... Federal Supply Schedules or regional term contracts. 101-25.110-2 Section 101-25.110-2 Public Contracts... REGULATIONS SUPPLY AND PROCUREMENT 25-GENERAL 25.1-General Policies § 101-25.110-2 Tires obtained through Federal Supply Schedules or regional term contracts. When tire manufacturers ship tires direct against...

  7. 41 CFR 101-25.110-2 - Tires obtained through Federal Supply Schedules or regional term contracts.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... Federal Supply Schedules or regional term contracts. 101-25.110-2 Section 101-25.110-2 Public Contracts... REGULATIONS SUPPLY AND PROCUREMENT 25-GENERAL 25.1-General Policies § 101-25.110-2 Tires obtained through Federal Supply Schedules or regional term contracts. When tire manufacturers ship tires direct against...

  8. Differential recognition of the ORF2 region in a complete genome sequence of porcine circovirus type 2 (PCV2) isolated from boar bone marrow in Korea.

    PubMed

    Kweon, Chang-Hee; Nguyen, Lien Thi Kim; Yoo, Mi-Sun; Kang, Seung-Won

    2015-09-15

    Porcine circovirus type 2 (PCV2) is the causative agent of post-weaning multisystemic wasting syndrome (PMWS) in swine. Here, a phylogenetic tree was constructed using PCV2 nucleotide sequences derived from the bone marrow of Korean boar and previously reported PCV2 sequences isolated from various countries. PCV2 from Korean boar bone marrow (KC188796) was classified into the group containing PCV2a-Canada and other PCV2 strain from Korea. While the ORF1 region of the PCV2 genome was highly conserved, ORF2 (the capsid protein coding region) was relatively variable. The nucleotide sequences for bone marrow-derived PCV2 were 93.4-99.0% homologous to the other reference sequences. The deduced amino acid sequences for the ORF1 and ORF2 coding regions were 97.4-99.3% and 84.5-97.4% homologous with the other reference strains, respectively, indicating that KC188796 did not differ markedly from the other PCV2 strains. Phylogenetic analysis demonstrated that bone marrow-derived PCV2 was highly similar to PCV2a from Canada and may be related to persistent PCV2 infections in swine. Copyright © 2015 Elsevier B.V. All rights reserved.

  9. Deletion and duplication within the p11.2 region of chromosome 17

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    McCorquodale, D.J.; McCorquodale, M.; Bereziouk, O.

    1994-09-01

    A 7 1/2-year-old male patient presented with mild mental retardation, speech delay, hyperactivity, behavioral problems, mild facial hypoplasia, short broad hands, digital anomalies, and self-injurious behavior. Chromosomes obtained from peripheral blood cells revealed a deletion of 17p11.2 in about 40% of the metaphases examined, suggesting that the patient had Smith-Magenis Syndrome. A similar pattern of mosaicism in peripheral blood cells, but not in fibroblasts in which all cells displayed the deletion, has been previously reported. Since some cases of Smith-Magenis Syndrome have a deletion that extends into the region associated with Charcot-Marie-Tooth (CMT) Syndrome, we examined interphase cells with amore » CMT1A-specific probe by the method of fluorescence in situ hybridization. The CMT1A region was not deleted, but about 40% of the cells gave signals indicating a duplication of the CMT1A region. The patient has not presented neuropathies associated with CMT at this time. Future tracking of the patient should be informative.« less

  10. Mua (HP0868) Is a Nickel-Binding Protein That Modulates Urease Activity in Helicobacter pylori

    PubMed Central

    Benoit, Stéphane L.; Maier, Robert J.

    2011-01-01

    A novel mechanism aimed at controlling urease expression in Helicobacter pylori in the presence of ample nickel is described. Higher urease activities were observed in an hp0868 mutant (than in the wild type) in cells supplemented with nickel, suggesting that the HP0868 protein (herein named Mua for modulator of urease activity) represses urease activity when nickel concentrations are ample. The increase in urease activity in the Δmua mutant was linked to an increase in urease transcription and synthesis, as shown by quantitative real-time PCR, SDS-PAGE, and immunoblotting against UreAB. Increased urease synthesis was also detected in a Δmua ΔnikR double mutant strain. The Δmua mutant was more sensitive to nickel toxicity but more resistant to acid challenge than was the wild-type strain. Pure Mua protein binds 2 moles of Ni2+ per mole of dimer. Electrophoretic mobility shift assays did not reveal any binding of Mua to the ureA promoter or other selected promoters (nikR, arsRS, 5′ ureB-sRNAp). Previous yeast two-hybrid studies indicated that Mua and RpoD may interact; however, only a weak interaction was detected via cross-linking with pure components and this could not be verified by another approach. There was no significant difference in the intracellular nickel level between wild-type and mua mutant cells. Taken together, our results suggest the HP0868 gene product represses urease transcription when nickel levels are high through an as-yet-uncharacterized mechanism, thus counterbalancing the well-described NikR-mediated activation. PMID:21505055

  11. Meta-analyses of structural regional cerebral effects in type 1 and type 2 diabetes.

    PubMed

    Moulton, Calum D; Costafreda, Sergi G; Horton, Paul; Ismail, Khalida; Fu, Cynthia H Y

    2015-12-01

    Diabetes is associated with macrovascular and microvascular complications and is a major risk factor for neurological and psychiatric disorders, such as dementia and depression. Type 1 diabetes (T1DM) and type 2 diabetes (T2DM) have distinct etiologies and pathophysiological effects while sharing a common endpoint of persistent hyperglycemia. Neuroimaging studies in T1DM have revealed reductions in numerous regions, including the parahippocampal and occipital regions, while in T2DM there have been numerous reports of hippocampal atrophy. This meta-analysis aimed to identify consistent regional abnormalities in cerebral structures in T1DM and T2DM respectively, and also to examine the impact of potential confounds, including age, depression and vascular risk factors. Neuroimaging studies of both voxel-based morphometry (VBM) data and volumetric data were included. Ten T1DM studies (n = 613 patients) and 23 T2DM studies (n = 1364 patients) fulfilled inclusion criteria. The T1DM meta-analysis revealed reduced bilateral thalamus grey matter density in adults. The T2DM meta-analysis revealed reduced global brain volume and regional atrophy in the hippocampi, basal ganglia, and orbitofrontal and occipital lobes. Moreover, hippocampal atrophy in T2DM was not modified by hypertension, although there were more marked reductions in younger patients relative to healthy controls. In conclusion, T1DM and T2DM demonstrated distinct cerebral effects with generalised and specific target areas of grey matter reduction. Thalamic atrophy in T1DM may be a substrate of associated cognitive deficits. In T2DM, global cerebral atrophy may reflect atherosclerotic factors, while hippocampal atrophy was an independent effect providing a potential common neuropathological etiology for the comorbidity of T2DM with dementia and depression.

  12. Correlation Between Presence of imaA (HP0289) and Inflammation in H. pylori Infected Patients through Analysis of Bacterial DNA

    NASA Astrophysics Data System (ADS)

    Gupta, A.; Jain, S.

    2012-12-01

    The bacterium Helicobacter pylori inhabits the stomachs of nearly half of the world's human population, yet only a small fraction (20%) of those people are harmfully affected by the organism. Inflammation caused by the species often results in stomach ulcers or even cancer in these infected patients. Previous studies indicate that the uncharacterized H. pylori gene imaA (HP0289) may be responsible for suppressing this inflammation. Correlation between the intactness of the gene and inflammation levels in patients was determined through analysis of 105 DNA samples from H. pylori infected patients. Traditional PCR and gel electrophoresis techniques were used in the experimentation process. Primers including AC235, 5'imaARev, 3'imaAFor2446, 3'imaARevClinical, ureA637For, ureA637Rev, interFor1, interRev1, interFor2, and interRev2 were used to identify deletions in HP0289 in each DNA sample. The results from this analysis could allow for eventual remediation of the adverse effects of H. pylori.

  13. Effects of the halophytes Tecticornia indica and Suaeda fruticosa on soil enzyme activities in a Mediterranean Sabkha.

    PubMed

    Ouni, Youssef; Lakhdar, Abdelbasset; Rabhi, Mokded; Smaoui, Abderrazak; Maria, A Rao; Chedly, Abdelly

    2013-01-01

    In the present work, we studied the effectiveness of the predominant halophytes of Soliman sabkha (Tecticornia indica and Suaeda fruticosa) to promote soil biological activities and ecosystem productivity. Soil Arylsulphatese ARY, beta-glucosidase beta-GLU, phosphatase PHO, invertase INV, urease URE, and dehydogenase DES activities in Extra- and Intra-tuft halophytes and plant productivity were assessed. Results revealed a high increase of microbial community and ARY, beta-GLU, PHO, INV URE and DES activities (+298%, +400%, +800%, +350%, +320%, +25% and +759%, respectively) in Intra-tuft rhizosphere as compared to Extra-tuft one, which is likely due to the significant decrease of salinity in the rhizosphere of Tecticornia indica and Suaeda fruticosa. Both perennial plants exhibited high productivities (7.4 t dry weight ha(-1) and 2.2 t dry weight ha(-1), respectively) and Na+-hyperaccumulating capacities (0.75 t Na+ ha(-1) and 0.22 t Na+ ha(-1), respectively), reducing salt constraint and favouring soil fertility. This constitutes a promising alternative to enhance productivity in such a salt-affected biotope by offering suitable microhabitat for annual glycophytes.

  14. A new analysis of Mars "Special Regions": findings of the Second MEPAG Special Regions Science Analysis Group (SR-SAG2)

    USGS Publications Warehouse

    Rummel, John D.; Beaty, David W.; Jones, Melissa A.; Bakermans, Corien; Barlow, Nadine G.; Boston, Penelope J.; Chevrier, Vincent F.; Clark, Benton C.; de Vera, Jean-Pierre P.; Gough, Raina V.; Hallsworth, John E.; Head, James W.; Hipkin, Victoria J.; Kieft, Thomas L.; McEwen, Alfred S.; Mellon, Michael T.; Mikucki, Jill A.; Nicholson, Wayne L.; Omelon, Christopher R.; Peterson, Ronald; Roden, Eric E.; Lollar, Barbara Sherwood; Tanaka, Kenneth L.; Viola, Donna; Wray, James J.

    2014-01-01

    A committee of the Mars Exploration Program Analysis Group (MEPAG) has reviewed and updated the description of Special Regions on Mars as places where terrestrial organisms might replicate (per the COSPAR Planetary Protection Policy). This review and update was conducted by an international team (SR-SAG2) drawn from both the biological science and Mars exploration communities, focused on understanding when and where Special Regions could occur. The study applied recently available data about martian environments and about terrestrial organisms, building on a previous analysis of Mars Special Regions (2006) undertaken by a similar team. Since then, a new body of highly relevant information has been generated from the Mars Reconnaissance Orbiter (launched in 2005) and Phoenix (2007) and data from Mars Express and the twin Mars Exploration Rovers (all 2003). Results have also been gleaned from the Mars Science Laboratory (launched in 2011). In addition to Mars data, there is a considerable body of new data regarding the known environmental limits to life on Earth—including the potential for terrestrial microbial life to survive and replicate under martian environmental conditions. The SR-SAG2 analysis has included an examination of new Mars models relevant to natural environmental variation in water activity and temperature; a review and reconsideration of the current parameters used to define Special Regions; and updated maps and descriptions of the martian environments recommended for treatment as "Uncertain" or "Special" as natural features or those potentially formed by the influence of future landed spacecraft. Significant changes in our knowledge of the capabilities of terrestrial organisms and the existence of possibly habitable martian environments have led to a new appreciation of where Mars Special Regions may be identified and protected. The SR-SAG also considered the impact of Special Regions on potential future human missions to Mars, both as locations

  15. A new analysis of Mars "Special Regions": findings of the second MEPAG Special Regions Science Analysis Group (SR-SAG2).

    PubMed

    Rummel, John D; Beaty, David W; Jones, Melissa A; Bakermans, Corien; Barlow, Nadine G; Boston, Penelope J; Chevrier, Vincent F; Clark, Benton C; de Vera, Jean-Pierre P; Gough, Raina V; Hallsworth, John E; Head, James W; Hipkin, Victoria J; Kieft, Thomas L; McEwen, Alfred S; Mellon, Michael T; Mikucki, Jill A; Nicholson, Wayne L; Omelon, Christopher R; Peterson, Ronald; Roden, Eric E; Sherwood Lollar, Barbara; Tanaka, Kenneth L; Viola, Donna; Wray, James J

    2014-11-01

    A committee of the Mars Exploration Program Analysis Group (MEPAG) has reviewed and updated the description of Special Regions on Mars as places where terrestrial organisms might replicate (per the COSPAR Planetary Protection Policy). This review and update was conducted by an international team (SR-SAG2) drawn from both the biological science and Mars exploration communities, focused on understanding when and where Special Regions could occur. The study applied recently available data about martian environments and about terrestrial organisms, building on a previous analysis of Mars Special Regions (2006) undertaken by a similar team. Since then, a new body of highly relevant information has been generated from the Mars Reconnaissance Orbiter (launched in 2005) and Phoenix (2007) and data from Mars Express and the twin Mars Exploration Rovers (all 2003). Results have also been gleaned from the Mars Science Laboratory (launched in 2011). In addition to Mars data, there is a considerable body of new data regarding the known environmental limits to life on Earth-including the potential for terrestrial microbial life to survive and replicate under martian environmental conditions. The SR-SAG2 analysis has included an examination of new Mars models relevant to natural environmental variation in water activity and temperature; a review and reconsideration of the current parameters used to define Special Regions; and updated maps and descriptions of the martian environments recommended for treatment as "Uncertain" or "Special" as natural features or those potentially formed by the influence of future landed spacecraft. Significant changes in our knowledge of the capabilities of terrestrial organisms and the existence of possibly habitable martian environments have led to a new appreciation of where Mars Special Regions may be identified and protected. The SR-SAG also considered the impact of Special Regions on potential future human missions to Mars, both as locations of

  16. Temporal and regional alterations in NMDA receptor expression in Mecp2-null mice

    PubMed Central

    Blue, Mary E.; Kaufmann, Walter E.; Bressler, Joseph; Eyring, Charlotte; O’Driscoll, Cliona; Naidu, SakkuBai; Johnston, Michael V.

    2014-01-01

    Our previous postmortem study of girls with Rett Syndrome (RTT), a development disorder caused by MECP2 mutations, found increases in the density of NMDA receptors in the prefrontal cortex of 2–8 year-old girls, while girls older than 10 years had reductions in NMDA receptors compared to age matched controls (Blue et al., 1999b). Using [3H]-CGP to label NMDA type glutamate receptors in 2 and 7 week old wildtype (WT), Mecp2-null and Mecp2-heterozygous (HET) mice (Bird model), we found that frontal areas of the brain also exhibited a bimodal pattern in NMDA expression, with increased densities of NMDA receptors in Mecp2-null mice at 2 weeks of age, but decreased densities at 7 weeks of age. Visual cortex showed a similar pattern, while other cortical regions only exhibited changes in NMDA receptor densities at 2 weeks (retrosplenial granular) or 7 weeks (somatosensory). In thalamus of null mice, NMDA receptors were increased at 2 and 7 weeks. No significant differences in density were found between HET and WT mice at both ages. Western blots for NMDAR1 expression in frontal brain showed higher levels of expression in Mecp2-null mice at two weeks of age, but not at 1 or 7 weeks of age. Our mouse data support the notion that deficient MeCP2 function is the primary cause of the NMDA receptor changes we observed in RTT. Furthermore, the findings of regional and temporal differences in NMDA expression illustrate the importance of age and brain region in evaluating different genotypes of mice. PMID:21901842

  17. Detecting fossil fuel emissions patterns from subcontinental regions using North American in situ CO2 measurements.

    PubMed

    Shiga, Yoichi P; Michalak, Anna M; Gourdji, Sharon M; Mueller, Kim L; Yadav, Vineet

    2014-06-28

    The ability to monitor fossil fuel carbon dioxide (FFCO 2 ) emissions from subcontinental regions using atmospheric CO 2 observations remains an important but unrealized goal. Here we explore a necessary but not sufficient component of this goal, namely, the basic question of the detectability of FFCO 2 emissions from subcontinental regions. Detectability is evaluated by examining the degree to which FFCO 2 emissions patterns from specific regions are needed to explain the variability observed in high-frequency atmospheric CO 2 observations. Analyses using a CO 2 monitoring network of 35 continuous measurement towers over North America show that FFCO 2 emissions are difficult to detect during nonwinter months. We find that the compounding effects of the seasonality of atmospheric transport patterns and the biospheric CO 2 flux signal dramatically hamper the detectability of FFCO 2 emissions. Results from several synthetic data case studies highlight the need for advancements in data coverage and transport model accuracy if the goal of atmospheric measurement-based FFCO 2 emissions detection and estimation is to be achieved beyond urban scales. Poor detectability of fossil fuel CO 2 emissions from subcontinental regionsDetectability assessed via attribution of emissions patterns in atmospheric dataLoss in detectability due to transport modeling errors and biospheric signal.

  18. Lidar Measurements of Atmospheric CO2 From Regional to Global Scales

    NASA Technical Reports Server (NTRS)

    Lin, Bing; Harrison, F. Wallace; Nehrir, Amin; Browell, Edward; Dobler, Jeremy; Campbell, Joel; Meadows, Byron; Obland, Michael; Ismail, Syed; Kooi, Susan; hide

    2015-01-01

    Atmospheric CO2 is a critical forcing for the Earth's climate and the knowledge on its distributions and variations influences predictions of the Earth's future climate. Large uncertainties in the predictions persist due to limited observations. This study uses the airborne Intensity-Modulated Continuous-Wave (IMCW) lidar developed at NASA Langley Research Center to measure regional atmospheric CO2 spatio-temporal variations. Further lidar development and demonstration will provide the capability of global atmospheric CO2 estimations from space, which will significantly advances our knowledge on atmospheric CO2 and reduce the uncertainties in the predictions of future climate. In this presentation, atmospheric CO2 column measurements from airborne flight campaigns and lidar system simulations for space missions will be discussed. A measurement precision of approx.0.3 ppmv for a 10-s average over desert and vegetated surfaces has been achieved. Data analysis also shows that airborne lidar CO2 column measurements over these surfaces agree well with in-situ measurements. Even when thin cirrus clouds present, consistent CO2 column measurements between clear and thin cirrus cloudy skies are obtained. Airborne flight campaigns have demonstrated that precise atmospheric column CO2 values can be measured from current IM-CW lidar systems, which will lead to use this airborne technique in monitoring CO2 sinks and sources in regional and continental scales as proposed by the NASA Atmospheric Carbon and Transport â€" America project. Furthermore, analyses of space CO2 measurements shows that applying the current IM-CW lidar technology and approach to space, the CO2 science goals of space missions will be achieved, and uncertainties in CO2 distributions and variations will be reduced.

  19. Aura OMI observations of changes in SO2 and NO2 emissions at local, regional and global scales

    NASA Astrophysics Data System (ADS)

    Krotkov, N. A.; McLinden, C. A.; Li, C.; Lamsal, L. N.; Celarier, E. A.; Marchenko, S. V.; Swartz, W.; Bucsela, E. J.; Joiner, J.; Duncan, B. N.; Boersma, K. F.; Veefkind, P.; Levelt, P.; Fioletov, V.; Dickerson, R. R.; He, H.; Lu, Z.; Streets, D. G.

    2015-12-01

    Space-based pollution monitoring from current and planned satellite UV-Vis spectrometers play an increasingly important role in studies of tropospheric chemistry and also air quality applications to help mitigate anthropogenic and natural impacts on sensitive ecosystems, and human health. We present long-term changes in tropospheric SO2 and NO2 over some of the most polluted industrialized regions of the world observed by the Ozone Monitoring Instrument (OMI) onboard NASA's Aura satellite. Using OMI data, we identified about 400 SO2 "hot spots" and estimated emissions from them. In many regions emissions and their ambient pollution levels have decreased significantly, such as over eastern US, Europe and China. OMI observed about 50% reduction in SO2 and NO2 pollution over the North China plain in 2012-2014 that can be attributed to both government efforts to restrain emissions from the power and industrial sectors and the economic slowdown. While much smaller, India's SO2 and NO2 emissions from coal power plants and smelters are growing at a fast pace, increasing by about 200% and 50% from 2005 to 2014. Over Europe and the US OMI-observed trends agree well with those from available in situ measurements of surface concentrations, deposition and emissions data. However, for some regions (e.g., Mexico, Middle East) the emission inventories may be incomplete and OMI can provide emission estimates for missing sources, such as SO2 sources observed over the Persian Gulf. It is essential to continue long-term overlapping satellite data records of air quality with increased spatial and temporal resolution to resolve point pollution sources using oversampling technique. We discuss how Aura OMI pollution measurements and emission estimates will be continued with the US JPSS and European Sentinel series for the next 20 years and further enhanced by the addition of three geostationary UV-VIS instruments.

  20. The UKC2 regional coupled prediction system

    NASA Astrophysics Data System (ADS)

    Castillo, Juan; Lewis, Huw; Graham, Jennifer; Saulter, Andrew; Arnold, Alex; Fallmann, Joachim; Martinez de la Torre, Alberto; Blyth, Eleanor; Bricheno, Lucy

    2017-04-01

    It is hypothesized that more accurate prediction and warning of natural hazards, such as of the impacts of severe weather through the environment, requires a more integrated approach to forecasting. This approach also delivers research benefits through providing tools with which to explore the known interactions and feedbacks between different physical and biogeochemical components of the environment across sky, sea and land. This hypothesis is being tested in a UK regional context at km-scale through the UK Environmental Prediction Project. This presentation will provide an introduction to the UKC2 UK Environmental Prediction research system. This incorporates models of the atmosphere (Met Office Unified Model), land surface (JULES), shelf-sea ocean (NEMO) and ocean waves (WAVEWATCH III). These components are coupled (via OASIS3-MCT libraries) at unprecedentedly high resolution across the UK and the wider north-west European regional domain. A research framework has been established to explore the representation of feedback processes in coupled and uncoupled modes, providing a unique new research tool for UK environmental science. The presentation will highlight work undertaken to review and improve the computational cost of running these systems for efficient research application. Research will be presented highlighting case study evaluation on the sensitivity of the ocean and surface waves to the representation of feedbacks to the atmosphere, and on the sensitivity of weather systems and boundary layer cloud development to the exchange of heat and momentum at the ocean surface modified through sea surface temperature and wave-induced roughness. The presentation will discuss plans for future development through UKC3 and beyond.

  1. Waterborne Polymeric Films.

    DTIC Science & Technology

    1979-12-01

    resin types: 1) acrylic latex, 2) acrylic solution polymer, 3) epoxy emulsions, 4) polyurethane, 5) butadiene elastomeric latex, 6) polyester/ alkyd , 7...emulsions and the class of polyester/ alkyd resins were evaluated only as network, crosslinked films. -53- j z I, ACRYLIC SOLUTIONLATEX URE THANE ACRYLIC ...amount of "plasticizing" monomer such as ethyl acrylate . 2.3.1.3 Aqueous Polyester Alkyd Resins As indicated in section 2.2.7 of this report,

  2. VLA HI Zeeman Observations of the Cygnus X Region: DR 22 And ON 2

    NASA Astrophysics Data System (ADS)

    Mayo, Elizabeth A.; Troland, T. H.

    2010-01-01

    The Very Large Array in Socorro, New Mexico has been used to study the Zeeman Effect in the 21cm HI line seen in absorption against radio sources in the Cygnus X region. Cygnus X is geometrically favorable for Zeeman effect observations as the region lies along the mean field direction of the diffuse interstellar medium (ISM) of the galaxy. We present observations of two compact HII regions within Cygnus X, DR 22 and ON 2. The data show magnetic field strengths of the order -80 μG toward DR 22 alone with no significant detections toward ON 2. This information is used to estimate the magnetic energy of the DR 22 star-forming cloud, and allows for a complete analysis of the energetics of the region revealing the role of the magnetic field. Support for this work was provided by the NSF PAARE program to South Carolina State University under award AST-0750814.

  3. Regional Educational Laboratory Electronic Network Phase 2 System

    NASA Technical Reports Server (NTRS)

    Cradler, John

    1995-01-01

    The Far West Laboratory in collaboration with the other regional educational laboratories is establishing a regionally coordinated telecommunication network to electronically interconnect each of the ten regional laboratories with educators and education stakeholders from the school to the state level. For the national distributed information database, each lab is working with mid-level networks to establish a common interface for networking throughout the country and include topics of importance to education reform as assessment and technology planning.

  4. 75 FR 38019 - Airworthiness Directives; Bombardier, Inc. Model CL-600-2C10 (Regional Jet Series 700 & 701...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-07-01

    ... DEPARTMENT OF TRANSPORTATION Federal Aviation Administration 14 CFR Part 39 [Docket No. FAA-2009... Directives; Bombardier, Inc. Model CL-600-2C10 (Regional Jet Series 700 & 701) Airplanes, Model CL-600-2D15 (Regional Jet Series 705) Airplanes, and Model CL-600-2D24 (Regional Jet Series 900) Airplanes Correction In...

  5. Towards a regional CO2 budget for New Zealand from atmospheric measurements and backward Lagrangian modeling

    NASA Astrophysics Data System (ADS)

    Steinkamp, K.; Mikaloff-Fletcher, S.; Brailsford, G. W.; Moore, S.

    2013-12-01

    Between 1990 and 2011, the reported average annual growth in total greenhouse gas emissions had been 1.0% for New Zealand, with emissions reaching 73 Mt CO2-e in 2011. At the same time the net emissions (total plus LULUCF) grew by 4.2% each year on average and reached 59 Mt CO2-e in 2011, according to the Ministry for the Environment. This implies a shrinking sink for greenhouse gases in areas of land use/ land use change and forests (LULUCF). The uptake of CO2 by forests is the largest contributor to this sink and, therefore, plays a crucial role in New Zealand's carbon budget. Yet, it is among the least well-known components. In this study, we aim to develop a regional atmosphere inversion system to estimate net CO2 uptake by land areas in 2011 and 2012. This will serve as an alternative to the bottom-up estimates outlined above. We use the UK Met Office's Lagrangian dispersion model NAME III to link CO2 measurements at stations directly to atmospheric transport and potential source regions at the surface. By running the model in backward mode, we identify the degree to which potential regional sources of CO2 contribute to observed mid-afternoon mixing ratios, i.e., the footprint of a station. Footprints are computed over 2011-2012 for three stations across New Zealand: Baring Head, Lauder and Rainbow Mountain. NAME III uses hourly meteorological input from the regional forecast model NZLAM-12 over a domain covering New Zealand and the Tasman Sea at a horizontal resolution of 12 km. The footprints are then used in a regional inversion to find the optimal distribution of CO2 sources and sinks, i.e., the one leading to the best match with the measurements at all stations. We present results from the footprint analysis and show that the three stations are sensitive to distinct source regions that do not overlap and, together, cover large parts of New Zealand. Hence, the data from the stations carry complementary information on CO2 sinks in sources throughout the

  6. Morphology of ionospheric F2 region variability associated with sudden stratospheric warmings

    NASA Astrophysics Data System (ADS)

    Gupta, Sumedha; Upadhayaya, A. K.

    2017-07-01

    The effect of sudden stratospheric warming (SSW) on the F2 region ionosphere has been extensively analyzed for the major event of year 2009, apart from a few reports on other major and minor events. Morphology of ionospheric responses during SSW can be better comprehended by analyzing such warming events under different solar, geomagnetic, and meteorological conditions. We investigate the features of F2 region variability following the SSW events of 2010, 2011, 2012, 2013, 2014, 2015, and 2016, using ionosonde data from the Asian region covering a broad latitudinal range from 26.6°N to 45.1°N. We find perceptible ionospheric variations in electron densities during these warming events which is accompanied by a large variation of 117% within enhancements, as compared to a meagre variation of 11% within depressions, during these events. We also examine 6 months data at these latitudes and longitudes and find that the maximum and minimum variations in F2 layer critical frequency are observed during each SSW period. The influence of quasi-stationary 16 day planetary waves is seen during these SSW events. Further, a recently proposed parameter "SSW integrated strength" by Vieira et al. (2017) to characterize SSW event with respect to ionosphere is also examined. It is seen that it does not fit well for these seven SSW events at these latitudes and longitudes.

  7. 2-D Path Corrections for Local and Regional Coda Waves: A Test of Transportability

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Mayeda, K M; Malagnini, L; Phillips, W S

    2005-07-13

    Reliable estimates of the seismic source spectrum are necessary for accurate magnitude, yield, and energy estimation. In particular, how seismic radiated energy scales with increasing earthquake size has been the focus of recent debate within the community and has direct implications on earthquake source physics studies as well as hazard mitigation. The 1-D coda methodology of Mayeda et al. [2003] has provided the lowest variance estimate of the source spectrum when compared against traditional approaches that use direct S-waves, thus making it ideal for networks that have sparse station distribution. The 1-D coda methodology has been mostly confined to regionsmore » of approximately uniform complexity. For larger, more geophysically complicated regions, 2-D path corrections may be required. We will compare performance of 1-D versus 2-D path corrections in a variety of regions. First, the complicated tectonics of the northern California region coupled with high quality broadband seismic data provides for an ideal ''apples-to-apples'' test of 1-D and 2-D path assumptions on direct waves and their coda. Next, we will compare results for the Italian Alps using high frequency data from the University of Genoa. For Northern California, we used the same station and event distribution and compared 1-D and 2-D path corrections and observed the following results: (1) 1-D coda results reduced the amplitude variance relative to direct S-waves by roughly a factor of 8 (800%); (2) Applying a 2-D correction to the coda resulted in up to 40% variance reduction from the 1-D coda results; (3) 2-D direct S-wave results, though better than 1-D direct waves, were significantly worse than the 1-D coda. We found that coda-based moment-rate source spectra derived from the 2-D approach were essentially identical to those from the 1-D approach for frequencies less than {approx}0.7-Hz, however for the high frequencies (0.7 {le} f {le} 8.0-Hz), the 2-D approach resulted in inter

  8. Emissions of air pollutants and greenhouse gases over Asian regions during 2000-2008: Regional Emission inventory in ASia (REAS) version 2

    NASA Astrophysics Data System (ADS)

    Kurokawa, J.; Ohara, T.; Morikawa, T.; Hanayama, S.; Janssens-Maenhout, G.; Fukui, T.; Kawashima, K.; Akimoto, H.

    2013-11-01

    We have updated the Regional Emission inventory in ASia (REAS) as version 2.1. REAS 2.1 includes most major air pollutants and greenhouse gases from each year during 2000 and 2008 and following areas of Asia: East, Southeast, South, and Central Asia and the Asian part of Russia. Emissions are estimated for each country and region using updated activity data and parameters. Monthly gridded data with a 0.25° × 0.25° resolution are also provided. Asian emissions for each species in 2008 are as follows (with their growth rate from 2000 to 2008): 56.9 Tg (+34%) for SO2, 53.9 Tg (+54%) for NOx, 359.5 Tg (+34%) for CO, 68.5 Tg (+46%) for non-methane volatile organic compounds, 32.8 Tg (+17%) for NH3, 36.4 Tg (+45%) for PM10, 24.7 Tg (+42%) for PM2.5, 3.03 Tg (+35%) for black carbon, 7.72 Tg (+21%) for organic carbon, 182.2 Tg (+32%) for CH4, 5.80 Tg (+18%) for N2O, and 16.0 Pg (+57%) for CO2. By country, China and India were respectively the largest and second largest contributors to Asian emissions. Both countries also had higher growth rates in emissions than others because of their continuous increases in energy consumption, industrial activities, and infrastructure development. In China, emission mitigation measures have been implemented gradually. Emissions of SO2 in China increased from 2000 to 2006 and then began to decrease as flue-gas desulphurization was installed to large power plants. On the other hand, emissions of air pollutants in total East Asia except for China decreased from 2000 to 2008 owing to lower economic growth rates and more effective emission regulations in Japan, South Korea, and Taiwan. Emissions from other regions generally increased from 2000 to 2008, although their relative shares of total Asian emissions are smaller than those of China and India. Tables of annual emissions by country and region broken down by sub-sector and fuel type, and monthly gridded emission data with a resolution of 0.25° × 0.25° for the major sectors are

  9. Emissions of air pollutants and greenhouse gases over Asian regions during 2000-2008: Regional Emission inventory in ASia (REAS) version 2

    NASA Astrophysics Data System (ADS)

    Kurokawa, J.; Ohara, T.; Morikawa, T.; Hanayama, S.; Greet, J.-M.; Fukui, T.; Kawashima, K.; Akimoto, H.

    2013-04-01

    We have updated the Regional Emission inventory in ASia (REAS) as version 2.1. REAS 2.1 includes most major air pollutants and greenhouse gases from each year during 2000 and 2008 and following areas of Asia: East, Southeast, South, and Central Asia and the Asian part of Russia. Emissions are estimated for each country and region using updated activity data and parameters. Monthly gridded data with a 0.25 × 0.25° resolution are also provided. Asian emissions for each species in 2008 are as follows (with their growth rate from 2000 to 2008): 56.9 Tg (+34%) for SO2, 53.9 Tg (+54%) for NOx, 359.5 Tg (+34%) for CO, 68.5 Tg (+46%) for non-methane volatile organic compounds, 32.8 Tg (+17%) for NH3, 36.4 Tg (+45%) for PM10, 24.7 Tg (+42%) for PM2.5, 3.03 Tg (+35%) for black carbon, 7.72 Tg (+21%) for organic carbon, 182.2 Tg (+32%) for CH4, 5.80 Tg (+18%) for N2O, and 16.7 Pg (+59%) for CO2. By country, China and India were respectively the largest and second largest contributors to Asian emissions. Both countries also had higher growth rates in emissions than others because of their continuous increases in energy consumption, industrial activities, and infrastructure development. In China, emission mitigation measures have been implemented gradually. Emissions of SO2 in China increased from 2000 to 2006 and then began to decrease as flue-gas desulfurization was installed to large power plants. On the other hand, emissions of air pollutants in total East Asia except for China decreased from 2000 to 2008 owing to lower economic growth rates and more effective emission regulations in Japan, South Korea, and Taiwan. Emissions from other regions generally increased from 2000 to 2008, although their relative shares of total Asian emissions are smaller than those of China and India. Tables of annual emissions by country and region broken down by sub-sector and fuel type, and monthly gridded emission data with a resolution of 0.25 × 0.25° for the major sectors are available

  10. RENU 2 UV Measurements of Atomic Oxygen in the Cusp Region

    NASA Astrophysics Data System (ADS)

    Fritz, B.; Lessard, M.; Paxton, L. J.; Cook, T.; Lynch, K. A.; Clemmons, J. H.; Hecht, J. H.; Hysell, D. L.; Crowley, G.

    2016-12-01

    The RENU 2 NASA sounding rocket mission launched from the Andoya Space Center on 13 December, 2015 into the dayside cusp region. A UV Photometer (UV PMT) provided by the University of New Hampshire was oriented to look up along the local magnetic field line as the payload passed through a poleward moving auroral form (PMAF). The bandpass filter on the UV PMT isolated emissions of atomic oxygen at both 130.4 nm and 135.6 nm. The instrument measured a clear enhancement in the topside ionosphere as the payload descended through a region of soft electron precipitation. The RENU 2 UV PMT was flown uncalibrated but measured a clear signal with both a major overall structure as well as several smaller peaks of fine structure. An identical spare has been built and calibrated using a Paresce UV light source at UMass-Lowell to compare and correlate with the flight data. An approximation of the flight data luminosity from the spare instrument and other flight data from RENU 2 is used in a radiative transport model to infer structure of upwelling neutral atomic oxygen above the PMAF.

  11. PMF and PSCF based source apportionment of PM2.5 at a regional background site in North China

    NASA Astrophysics Data System (ADS)

    Zong, Zheng; Wang, Xiaoping; Tian, Chongguo; Chen, Yingjun; Fu, Shanfei; Qu, Lin; Ji, Ling; Li, Jun; Zhang, Gan

    2018-05-01

    To apportion regional PM2.5 (atmospheric particles with aerodynamic diameter < 2.5 μm) source types and their geographic pattern in North China, 120 daily PM2.5 samples on Beihuangcheng Island (BH, a regional background site in North China) were collected from August 20th, 2014 to September 15th, 2015 showing one-year period. After the chemical analyses on carbonaceous species, water-soluble ions and inorganic elements, various approaches, such as Mann-Kendall test, chemical mass closure, ISORROPIA II model, Positive Matrix Factorization (PMF) linked with Potential Source Contribution Function (PSCF), were used to explore the PM2.5 speciation, sources, and source regions. Consequently, distinct seasonal variations of PM2.5 and its main species were found and could be explained by varying emission source characteristics. Based on PMF model, seven source factors for PM2.5 were identified, which were coal combustion + biomass burning, vehicle emission, mineral dust, ship emission, sea salt, industry source, refined chrome industry with the contribution of 48.21%, 30.33%, 7.24%, 6.63%, 3.51%, 3.2%, and 0.88%, respectively. In addition, PSCF analysis using the daily contribution of each factor from PMF result suggested that Shandong peninsula and Hebei province were identified as the high potential region for coal combustion + biomass burning; Beijing-Tianjin-Hebei (BTH) region was the main source region for industry source; Bohai Sea and East China Sea were found to be of high source potential for ship emission; Geographical region located northwest of BH Island was possessed of high probability for sea salt; Mineral dust presumably came from the region of Mongolia; Refined chrome industry mostly came from Liaoning, Jilin province; The vehicle emission was primarily of BTH region origin, centring on metropolises, such as Beijing and Tianjin. These results provided precious implications for PM2.5 control strategies in North China.

  12. Serologic screening for 13 infectious agents in roe deer (Capreolus capreolus) in Flanders

    PubMed Central

    Tavernier, Paul; Sys, Stanislas U.; De Clercq, Kris; De Leeuw, Ilse; Caij, Anne Brigitte; De Baere, Miet; De Regge, Nick; Fretin, David; Roupie, Virginie; Govaerts, Marc; Heyman, Paul; Vanrompay, Daisy; Yin, Lizi; Kalmar, Isabelle; Suin, Vanessa; Brochier, Bernard; Dobly, Alexandre; De Craeye, Stéphane; Roelandt, Sophie; Goossens, Els; Roels, Stefan

    2015-01-01

    Introduction In order to investigate the role of roe deer in the maintenance and transmission of infectious animal and human diseases in Flanders, we conducted a serologic screening in 12 hunting areas. Materials and methods Roe deer sera collected between 2008 and 2013 (n=190) were examined for antibodies against 13 infectious agents, using indirect enzyme-linked immunosorbent assay, virus neutralisation, immunofluorescence, or microagglutination test, depending on the agent. Results and discussion High numbers of seropositives were found for Anaplasma phagocytophilum (45.8%), Toxoplasma gondii (43.2%) and Schmallenberg virus (27.9%), the latter with a distinct temporal distribution pattern following the outbreak in domestic ruminants. Lower antibody prevalence was found for Chlamydia abortus (6.7%), tick-borne encephalitis virus (5.1%), Neospora caninum (4.8%), and Mycobacterium avium subsp paratuberculosis (4.1%). The lowest prevalences were found for Leptospira (1.7%), bovine viral diarrhoea virus 1 (1.3%), and Coxiella burnetii (1.2%). No antibodies were found against Brucella sp., bovine herpesvirus 1, and bluetongue virus. A significant difference in seroprevalence between ages (higher in adults >1 year) was found for N. caninum. Four doubtful reacting sera accounted for a significant difference in seroprevalence between sexes for C. abortus (higher in females). Conclusions Despite the more intensive landscape use in Flanders, the results are consistent with other European studies. Apart from maintaining C. abortus and MAP, roe deer do not seem to play an important role in the epidemiology of the examined zoonotic and domestic animal pathogens. Nevertheless, their meaning as sentinels should not be neglected in the absence of other wild cervid species. PMID:26609692

  13. Annual variation in event-scale precipitation δ2H at Barrow, AK, reflects vapor source region

    NASA Astrophysics Data System (ADS)

    Putman, Annie L.; Feng, Xiahong; Sonder, Leslie J.; Posmentier, Eric S.

    2017-04-01

    In this study, precipitation isotopic variations at Barrow, AK, USA, are linked to conditions at the moisture source region, along the transport path, and at the precipitation site. Seventy precipitation events between January 2009 and March 2013 were analyzed for δ2H and deuterium excess. For each precipitation event, vapor source regions were identified with the hybrid single-particle Lagrangian integrated trajectory (HYSPLIT) air parcel tracking program in back-cast mode. The results show that the vapor source region migrated annually, with the most distal (proximal) and southerly (northerly) vapor source regions occurring during the winter (summer). This may be related to equatorial expansion and poleward contraction of the polar circulation cell and the extent of Arctic sea ice cover. Annual cycles of vapor source region latitude and δ2H in precipitation were in phase; depleted (enriched) δ2H values were associated with winter (summer) and distal (proximal) vapor source regions. Precipitation δ2H responded to variation in vapor source region as reflected by significant correlations between δ2H with the following three parameters: (1) total cooling between lifted condensation level (LCL) and precipitating cloud at Barrow, ΔTcool, (2) meteorological conditions at the evaporation site quantified by 2 m dew point, Td, and (3) whether the vapor transport path crossed the Brooks and/or Alaskan ranges, expressed as a Boolean variable, mtn. These three variables explained 54 % of the variance (p<0. 001) in precipitation δ2H with a sensitivity of -3.51 ± 0.55 ‰ °C-1 (p<0. 001) to ΔTcool, 3.23 ± 0.83 ‰ °C-1 (p<0. 001) to Td, and -32.11 ± 11.04 ‰ (p = 0. 0049) depletion when mtn is true. The magnitude of each effect on isotopic composition also varied with vapor source region proximity. For storms with proximal vapor source regions

  14. The Ratio of Ortho- to Para-H2 in Photodissociation Regions

    NASA Technical Reports Server (NTRS)

    Sternberg, Amiel; Neufeld, David A.

    1999-01-01

    We discuss the ratio of ortho- to para-H2 in photodissociation regions (PDRs). We draw attention to an apparent confusion in the literature between the ortho-to-para ratio of molecules in FUV-pumped vibrationally excited states and the total H2 ortho-to-para abundance ratio. These ratios are not the same because the process of FUV pumping of fluorescent H2 emission in PDRs occurs via optically thick absorption lines. Thus gas with an equilibrium ratio of ortho- to para-H2 equal to 3 will yield FUV-pumped vibrationally excited ortho-to-para ratios smaller than 3, because the ortho-H2 pumping rates are preferentially reduced by optical depth effects. Indeed, if the ortho and para pumping lines are on the "square root" part of the curve of growth, then the expected ratio of ortho and para vibrational line strengths is 3(sup 1/2) approximately 1.7, close to the typically observed value. Thus, contrary to what has sometimes been stated in the literature, most previous measurements of the ratio of ortho- to para-H2 in vibrationally excited states are entirely consistent with a total ortho-to-para ratio of 3, the equilibrium value for temperatures greater than 200 K. We present an analysis and several detailed models that illustrate the relationship between the total ratios of ortho- to para-H2 and the vibrationally excited ortho-to-para ratios in PDRs. Recent Infrared Space Observatory measurements of pure rotational and vibrational H2 emissions from the PDR in the star-forming region S140 provide strong observational support for our conclusions.

  15. CO2-MEGAPARIS: Quantification of CO2 emissions from Paris megacity and their spread out to the neightbouring Centre region. (Invited)

    NASA Astrophysics Data System (ADS)

    Xueref-Remy, I.

    2010-12-01

    Atmospheric CO2 concentration has been increasing of more than 30% since the pre-industrial era due to human activities, and is very likely involved in the recent global temperature increase [IPCC, 2007]. Although we have good estimates of the CO2 fluxes on a global basis, and have a relatively well-established system to detect the large-scale trends, regional information (10-500km) is needed if society is ever to manage or verify carbon emissions. We must improve our understanding of regional variations in the sources and sinks of CO2 because they help identify possible sequestration or emission management options. New programs are needed to improve our understanding of meso-scale carbon fluxes, and to discriminate between the anthropogenic and biospheric sources which are very strongly overlapped in European countries. In this context we need to monitor the emissions originating from the megalopolis such as Paris and its agglomeration, and the way they are spreading in the background atmosphere. Nowadays, inventories (CITEPA, AIRPARIF) based on statistical information provide CO2 emissions from Ile de France and all others regions of France, but no independent verification based on CO2 measurements has been done yet. Atmospheric measurements coupled to a meso-scale model can be used to provide such verification, especially to detect the interannual and decadal trends which could result from regional management strategy. The CO2-MEGAPARIS project (2009-2012) objective is to develop four independent methods to verify the emission inventories, and to monitor the daily to monthly CO2 emissions from Ile de France as well as their spreading to neighbouring regions with a scale up to 2x2 km2. The first method consists in developing a synergy between a mesoscale model (CHIMERE/MM5), inventories and observations using a top-down approach based on an inversion technique to retrieve surface fluxes (3 new observing stations are developed among which the top of the Eiffel

  16. Source sector and region contributions to BC and PM 2.5 in Central Asia

    DOE PAGES

    Kulkarni, S.; Sobhani, N.; Miller-Schulze, J. P.; ...

    2015-02-18

    Particulate matter (PM) mass concentrations, seasonal cycles, source sector, and source region contributions in Central Asia (CA) are analyzed for the period April 2008–July 2009 using the Sulfur Transport and dEposition Model (STEM) chemical transport model and modeled meteorology from the Weather Research and Forecasting (WRF) model. Predicted aerosol optical depth (AOD) values (annual mean value ~0.2) in CA vary seasonally, with lowest values in the winter. Surface PM 2.5 concentrations (annual mean value ~10 μg m −3) also exhibit a seasonal cycle, with peak values and largest variability in the spring/summer, and lowest values and variability in the wintermore » (hourly values from 2 to 90 μg m −3). Surface concentrations of black carbon (BC) (mean value ~0.1 μg m −3) show peak values in the winter. The simulated values are compared to surface measurements of AOD as well as PM 2.5, PM 10, BC, and organic carbon (OC) mass concentrations at two regional sites in Kyrgyzstan (Lidar Station Teplokluchenka (LST) and Bishkek). The predicted values of AOD and PM mass concentrations and their seasonal cycles are fairly well captured. The carbonaceous aerosols are underpredicted in winter, and analysis suggests that the winter heating emissions are underestimated in the current inventory. Dust, from sources within and outside CA, is a significant component of the PM mass and drives the seasonal cycles of PM and AOD. On an annual basis, the power and industrial sectors are found to be the most important contributors to the anthropogenic portion of PM 2.5. Residential combustion and transportation are shown to be the most important sectors for BC. Biomass burning within and outside the region also contributes to elevated PM and BC concentrations. The analysis of the transport pathways and the variations in particulate matter mass and composition in CA demonstrates that this region is strategically located to characterize regional and intercontinental transport

  17. Prevalence and spatial distribution of bovine brucellosis in San Luis and La Pampa, Argentina.

    PubMed

    Aznar, M N; Linares, F J; Cosentino, B; Sago, A; La Sala, L; León, E; Duffy, S; Perez, A

    2015-08-15

    Bovine brucellosis (BB) is a zoonotic disease caused by Brucella abortus. BB is endemic in Argentina, where vaccination with Brucella abortus strain 19 is compulsory for 3-to-8 month-old heifers. The objectives of this study were to quantify the prevalence of BB and to identify factors associated with its occurrence, along with the spatial distribution of the disease, in the provinces of La Pampa and San Luis. A two-stage random sampling design was used to sample 8,965 cows (3,513 in La Pampa and 5,452 in San Luis) from 451 farms (187 in La Pampa and 264 in San Luis). Cow and herd prevalence were 1.8 % (95 % CI: 1.3-2.2; n = 157) and 19.7 % (95 % CI: 17.0-22.4; n = 89), respectively. Both cow-level and herd-level prevalence in La Pampa (2.4 and 26.0 %, respectively) were significantly higher than in San Luis (1.4 and 15.5 %, respectively). There were not differences between the proportions of reactive cattle compared to that obtained in a survey conducted in 2005. However, herd prevalence in La Pampa was significantly (P < 0.05) higher compared to that study. Disease was found to be spatially clustered in west La Pampa. The lower the bovine density and the calf/cow ratio, the higher odds of belonging to the cluster. The increase of farm prevalence in the last five years suggests that the disease is spreading and that control measures should be applied in the region. The cluster of infected farms was located in the west region of La Pampa. There, farms have lower animal densities and smaller cow/calf indices compared to the rest of the province. Although western La Pampa has more infected herds, within-farm prevalence was not higher, which suggests that the control program has been relatively successful in controlling the disease at the farm level, and/or that low animal density inherently results in low disease prevalence. Our results provide baseline information on the epidemiology of BB and its potential pattern of transmission in Argentina, which will ultimately

  18. Silencing NKD2 by Promoter Region Hypermethylation Promotes Esophageal Cancer Progression by Activating Wnt Signaling.

    PubMed

    Cao, Baoping; Yang, Weili; Jin, Yongshuai; Zhang, Meiying; He, Tao; Zhan, Qimin; Herman, James G; Zhong, Guanglin; Guo, Mingzhou

    2016-11-01

    Naked cuticle homolog 2 (NKD2) was found to be frequently methylated in human breast and gastric cancers. However, the epigenetic changes and mechanisms of NKD2 in human esophageal cancer remain unclear. Nine esophageal cancer cell lines and 154 cases of primary esophageal cancer samples were analyzed using methylation-specific polymerase chain reaction, immunohistochemical analysis, Western blot, and xenograft mouse models. Loss of NKD2 expression and complete methylation were found in KYSE150 and TE1 cells. Reduced NKD2 expression and partial methylation of the promoter region were observed in KYSE30, KYSE70, KYSE410, KYSE140, and COLO680 cells. High levels of NKD2 expression and unmethylation were detected in KYSE450 and TE8 cells. Reexpression of NKD2 was induced by 5-aza-2'-deoxycytidine in cells in which NKD2 was not expressed or cells in which NKD2 expression was reduced. NKD2 was methylated in 53.2% of human primary esophageal cancer samples (82 of 154), and promoter region hypermethylation was significantly associated with reduced expression of NKD2 (p < 0.01). NKD2 methylation was associated with tumor, node, and metastasis stage and lymph node metastasis (p < 0.01). Our results suggest that NKD2 is regulated by promoter region methylation and that methylation of NKD2 may serve as a prognostic marker in esophageal cancer. Our further studies demonstrate that NKD2 suppresses cell proliferation, colony formation, cell invasion, and migration and also induces G1/S checkpoint arrest in esophageal cancer cells. NKD2 suppressed xenograft tumor growth and inhibited Wnt signaling in human esophageal cancer cells. NKD2 is frequently methylated in human esophageal cancer, and the expression of NKD2 is regulated by promoter region methylation. NKD2 suppresses esophageal cancer progression by inhibiting Wnt signaling both in vitro and in vivo. Copyright © 2016 International Association for the Study of Lung Cancer. Published by Elsevier Inc. All rights reserved.

  19. Characterisation of regional skin temperatures in recreational surfers wearing a 2-mm wetsuit.

    PubMed

    Corona, Luis J; Simmons, Grant H; Nessler, Jeff A; Newcomer, Sean C

    2018-05-01

    The purpose of this study was to investigate skin temperatures across surfers' bodies while wearing a wetsuit during recreational surfing. Forty-six male recreational surfers participated in this study. Participants were instrumented with eight wireless iButton thermal sensors for the measurement of skin temperature, a Polar RCX5 heart rate monitor and a 2-mm full wetsuit. Following instrumentation, participants were instructed to engage in recreational surfing activities as normal. Significant differences (p < 0.001) in skin temperature (T sk ) were found across the body while wearing a wetsuit during recreational surfing. In addition, regional skin temperature changed across the session for several regions of the body (p < 0.001), and the magnitude of these changes varied significantly between regions. We show for the first time that significant differences exist in skin temperature across the body while wearing a wetsuit during a typical recreational surfing session. These findings may have implications for future wetsuit design. Practitioner Summary: This study investigated the impact of wearing a wetsuit during recreational surfing on regional skin temperatures. Results from this study suggest that skin temperatures differ significantly across the body while wearing a 2-mm wetsuit during recreational surfing. These findings may have implications for future wetsuit design.

  20. Genetic map of the spinocerebellar ataxia type 2 (SCA2) region on chromosome 12

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Nechiporuk, A.; Frederick, T.; Pulst, S.M.

    1994-09-01

    The autosomal dominant ataxias (SCAs) are a clinically and genetically heterogeneous group of neurodegenerative diseases characterized by progressive ataxia. At least four gene loci have been identified: SCA1 on chromosome (CHR) 6, SCA2 on CHR12, Machado-Joseph disease on CHR14, and SCA families that are not linked to any of the above loci. In addition, the gene causing dentato-rubro-pallido-luysian atrophy has been identified as an expanded CAG repeat on CHR 12p. As a necessary step in identifying the gene for SCA2, we now identified closer flanking markers. To do this we ordered microsatellite markers in the now identified closer flanking markers.more » To do this we ordered microsatellite markers in the region and then determined pairwise and multipoint lod scores between the markers and SCA2 in three large pedigrees with SCA. The following order was established with odds > 1,000:1 using six non-SCA pedigrees: D12S101-7.1cM-D12S58-0cM-IGF1-3.6cM-D12S78-1.4cM-D12S317-3.7cM-D12S84-0cM-D12S105-7.2cM-D12S79-7.0cM-PLA2. Using this ordered set of markers we examined linkage to SCA2 in three pedigrees of Italian, Austrian and French-Canadian descent. Pairwise linkage analysis resulted in significant positive lod scores for all markers. The highest pairwise lod score was obtained with D12S84/D12S105 (Z{sub max}=7.98, theta{sub max}=0.05). To further define the location of SCA2, we performed multipoint linkage analysis using the genetic map established above. The highest location score was obtained between D12S317 and D12S84/D12S105. A location of SCA2 between these loci was favored with odds > 100:1. These data likely narrow the SCA2 candidate region to approximately 3.7 cM. The relatively large large number of markers tightly linked to SCA2 will facilitate the assignment of additional SCA pedigrees to CHR12, and will help in the presymptomatic diagnosis of individuals in families with proven linkage to CHR12.« less

  1. The minimal promoter region of the dense-core vesicle protein IA-2: transcriptional regulation by CREB.

    PubMed

    Cai, Tao; Hirai, Hiroki; Xu, Huanyu; Notkins, Abner L

    2015-06-01

    IA-2 is a transmembrane protein found in the dense-core vesicles (DCV) of neuroendocrine cells and one of the major autoantigens in type 1 diabetes. DCV are involved in the secretion of hormones (e.g., insulin) and neurotransmitters. Stimulation of pancreatic β cells with glucose upregulates the expression of IA-2 and an increase in IA-2 results in an increase in the number of DCV. Little is known, however, about the promoter region of IA-2 or the transcriptional factors that regulate the expression of this gene. In the present study, we constructed eight deletion fragments from the upstream region of the IA-2 transcription start site and linked them to a luciferase reporter. By this approach, we have identified a short bp region (-216 to +115) that has strong promoter activity. We also identified a transcription factor, cAMP responsive element-binding protein (CREB), which binds to two CREB-related binding sites located in this region. The binding of CREB to these sites enhanced IA-2 transcription by more than fivefold. We confirmed these findings by site-directed mutagenesis, chromatin immunoprecipitation assays and RNAi inhibition. Based on these findings, we conclude that the PKA pathway is a critical, but not the exclusive signaling pathway involved in IA-2 gene expression.

  2. Isolation and identification of bovine Brucella isolates from Pakistan by biochemical tests and PCR.

    PubMed

    Ali, Shahzad; Ali, Qurban; Melzer, Falk; Khan, Iahtasham; Akhter, Shamim; Neubauer, Heinrich; Jamal, Syed M

    2014-01-01

    Brucellosis is endemic in bovines in Pakistan. The Brucella species and biovars involved, however, are unknown. The objectives of the present study were to isolate and characterize brucellae from seropositive milk samples, aborted fetuses, and vaginal swabs of cattle and buffaloes which had recently aborted. The seropositive milk samples, aborted fetuses, and vaginal swabs of cattle and buffaloes were collected from the Potohar Plateau, Pakistan. Isolation of brucellae was done on modified Farrell's serum dextrose agar. Isolates were characterized by conventional biotyping methods, while molecular typing was done by genus (B4/B5) and species-specific (Brucella abortus, Brucella melitensis, Brucella ovis, and Brucella suis) polymerase chain reaction (PCR). A total of 30 isolates were recovered from milk (n = 5), aborted fetuses (n = 13), and vaginal swabs (n = 12). Most isolates were from cattle (56.7 %). All of them were identified as B. abortus biovar 1 based on conventional biotyping methods and genus and species-specific PCR. This preliminary study provides the first report on the prevalence of B. abortus biovar 1 in cattle and buffaloes in Pakistan.

  3. Regional climatic effects of atmospheric SO2 on Mars

    NASA Technical Reports Server (NTRS)

    Postawko, S. E.; Fanale, F. P.

    1992-01-01

    The conditions under which the valley networks on Mars may have formed remains controversial. The magnitude of an atmospheric greenhouse effect by an early massive CO2 atmosphere has recently been questioned by Kasting. Recent calculations indicate that if solar luminosity were less than about 86 percent of its current value, formation of CO2 clouds in the Martian atmosphere would depress the atmospheric lapse rate and reduce the magnitude of surface warming. In light of recent revisions of magma generation on Mars during each Martian epoch, and the suggestions by Wanke et al. that the role of liquid SO2 should be more carefully explored, we have recalculated the potential greenhouse warming by atmospheric SO2 on Mars, with an emphasis on more localized effects. In the vicinity of an active eruption, the concentration of atmospheric SO2 will be higher than if it is assumed that the erupted SO2 is instantaneously globally distributed. The local steady-state concentration of SO2 is a function of the rate at which it is released, its atmospheric lifetime, and the rate at which local winds act to disperse the SO2. We have made estimates of eruption rates, length of eruption, and dispersion rates of volcanically released SO2, for a variety of atmospheric conditions and atmospheric lifetimes of SO2 to explore the maximum regional climatic effect of SO2.

  4. 12-Digit Watershed Boundary Data 1:24,000 for EPA Region 2 and Surrounding States (NAT_HYDROLOGY.HUC12_NRCS_REG2)

    EPA Pesticide Factsheets

    12 digit Hydrologic Units (HUCs) for EPA Region 2 and surrounding states (Northeastern states, parts of the Great Lakes, Puerto Rico and the USVI) downloaded from the Natural Resources Conservation Service (NRCS) Geospatial Gateway and imported into the EPA Region 2 Oracle/SDE database. This layer reflects 2009 updates to the national Watershed Boundary Database (WBD) that included new boundary data for New York and New Jersey.

  5. Application of the Ribosomal DNA ITS2 Region of Physalis (Solanaceae): DNA Barcoding and Phylogenetic Study

    PubMed Central

    Feng, Shangguo; Jiang, Mengying; Shi, Yujun; Jiao, Kaili; Shen, Chenjia; Lu, Jiangjie; Ying, Qicai; Wang, Huizhong

    2016-01-01

    Recently, commercial interest in Physalis species has grown worldwide due to their high nutritional value, edible fruit, and potential medicinal properties. However, many Physalis species have similar shapes and are easily confused, and consequently the phylogenetic relationships between Physalis species are poorly understood. This hinders their safe utilization and genetic resource conservation. In this study, the nuclear ribosomal ITS2 region was used to identify species and phylogenetically examine Physalis. Eighty-six ITS2 regions from 45 Physalis species were analyzed. The ITS2 sequences were aligned using Clustal W and genetic distances were calculated using MEGA V6.0. The results showed that ITS2 regions have significant intra- and inter-specific divergences, obvious barcoding gaps, and higher species discrimination rates (82.2% for both the BLASTA1 and nearest distance methods). In addition, the secondary structure of ITS2 provided another way to differentiate species. Cluster analysis based on ITS2 regions largely concurred with the relationships among Physalis species established by many previous molecular analyses, and showed that most sections of Physalis appear to be polyphyletic. Our results demonstrated that ITS2 can be used as an efficient and powerful marker in the identification and phylogenetic study of Physalis species. The technique provides a scientific basis for the conservation of Physalis plants and for utilization of resources. PMID:27486467

  6. A survey of brucellosis and tuberculosis in bison in and around Wood Buffalo National Park, Canada

    PubMed Central

    Tessaro, Stacy V.; Forbes, Lorry B.; Turcotte, Claude

    1990-01-01

    Examinations of complete or partial remains of 72 bison found dead in and around Wood Buffalo National Park, Canada, revealed evidence of brucellosis in 18 (25%) and tuberculosis in 15 (21%), with a combined prevalence of 42%. Urease-positive and ureasenegative strains of Brucella abortus biovar 1, and strains of biovar 2, were isolated from tissues of bison, including synovium and exudate from severe arthritic lesions. Mycobacterium bovis was isolated from a range of granulomatous lesions that were similar to those reported in tuberculous cattle. Diseased bison had a broad geographical distribution, and were found outside the park on at least three natural corridors. The diseases have a deleterious effect on this population of bison, and pose a health risk to other bison herds, livestock, and native hunters in the region. ImagesFigure 3.Figure 4. PMID:17423532

  7. Seasonal Variations of Atmospheric CO2 over Fire Affected Regions Based on GOSAT Observations

    NASA Astrophysics Data System (ADS)

    Shi, Y.; Matsunaga, T.

    2016-12-01

    Abstract: The carbon dioxide (CO2) emissions released from biomass burning significantly affect the temporal variations of atmospheric CO2 concentrations. Based on a long-term (July 2009-June 2015) retrieved datasets by the Greenhouse Gases Observing Satellite (GOSAT), the seasonal cycle and interannual variations of column-averaged volume mixing ratios of atmospheric carbon dioxide (XCO2) in four fire affected continental regions were investigated. The results showed Northern Africa had the largest seasonal variations after removing its regional long-term trend of XCO2 with peak-to-peak amplitude of 6.2 ppm within the year, higher than central South America (2.4 ppm), Southern Africa (3.8 ppm) and Australia (1.7 ppm). The detrended regional XCO2 was found to be positively correlated with the fire CO2 emissions during fire activity period and negatively correlated with vegetation photosynthesis activity with different seasonal variabilities. Northern Africa recorded the largest change of seasonal variations of detrended XCO2 with a total of 12.8 ppm during fire seasons, higher than central South America, Southern Africa and Australia with 5.4 ppm, 6.7 ppm and 2.2 ppm, respectively. During fire episode, the positive detrended XCO2 was noticed during June-November in central South America, December-June in Northern Africa, May-November in Southern Africa. The Pearson correlation coefficients between the variations of detrended XCO2 and fire CO2 emissions from GFED4 (Global Fire Emissions Database v4) achieved best correlations in Southern Africa (R=0.77, p<0.05). Meanwhile, Southern Africa also experienced a significant negative relationship between the variations of detrended XCO2 and vegetation activity (R=-0.84, p<0.05). This study revealed that fire CO2 emissions and vegetation activity contributed greatly to the seasonal variations of GOSAT XCO2 dataset.

  8. A framework for delineating the regional boundaries of PM2.5 pollution: A case study of China.

    PubMed

    Liu, Jianzheng; Li, Weifeng; Wu, Jiansheng

    2018-04-01

    Fine particulate matter (PM 2.5 ) pollution has been a major issue in many countries. Considerable studies have demonstrated that PM 2.5 pollution is a regional issue, but little research has been done to investigate the regional extent of PM 2.5 pollution or to define areas in which PM 2.5 pollutants interact. To allow for a better understanding of the regional nature and spatial patterns of PM 2.5 pollution, This study proposes a novel framework for delineating regional boundaries of PM 2.5 pollution. The framework consists of four steps, including cross-correlation analysis, time-series clustering, generation of Voronoi polygons, and polygon smoothing using polynomial approximation with exponential kernel method. Using the framework, the regional PM 2.5 boundaries for China are produced and the boundaries define areas where the monthly PM 2.5 time series of any two cities show, on average, more than 50% similarity with each other. These areas demonstrate straightforwardly that PM 2.5 pollution is not limited to a single city or a single province. We also found that the PM 2.5 areas in China tend to be larger in cold months, but more fragmented in warm months, suggesting that, in cold months, the interactions between PM 2.5 concentrations in adjacent cities are stronger than in warmer months. The proposed framework provides a tool to delineate PM 2.5 boundaries and identify areas where PM 2.5 pollutants interact. It can help define air pollution management zones and assess impacts related to PM 2.5 pollution. It can also be used in analyses of other air pollutants. Copyright © 2017 Elsevier Ltd. All rights reserved.

  9. The fast oxidation of SO2 in oil sands regions of Alberta,Canada

    NASA Astrophysics Data System (ADS)

    Amiri, N.; Norman, A. L.

    2016-12-01

    Secondary aerosols in the atmosphere play a significant role in the Earth's radiation budget and in human health. It is important to understand how secondary aerosols are formed. Atmospheric SO2 oxidation leads to secondary sulfate aerosols. The SO2 oxidation rate needs to be well defined to better understand aerosols and their effects and oxidation varies depending on the oxidants present. This research presents the results of a field campaign from 13 Aug to 5 Sep 2013 at the Wood Buffalo Air Monitoring Station 13 (AMS13) site just south of Fort MacKay, in which two lines of evidence show fast oxidation of SO2 in the region. Size-segregated sulfate aerosols and SO2 gas were collected on microfiber glass filters and filters treated by K2CO3 and glycerin respectively. The sulfur isotopic composition of sulfate aerosols and SO2 were measured. Periods when a nearby instrument was in operation (20m away), displayed markedly distinct d34S values from periods when it was not operational. The nearby instrument used enriched 34SO2, and this affected the resulting d34S values for all sulfate size fractions but not SO2 from our high volume sampler. The most pronounced contamination was observed for sulfate aerosols D<490nm, which are expected to be derived mostly from secondary sulfate. Furthermore, the concentration of SO2 collected on the high volume filters was significantly lower than the concentration measured by a co-located optical analyzer (m=0.4). These observations show that the isotopically enriched SO2 can be used as an unintentional ambient tracer experiment in the Oil Sands region, and that this SO2 was oxidized before reaching the high volume sampler. The results from our study show that SO2 oxidation in the Oil Sands regions in the presence of pollutants such as hydrocarbons is rapid.

  10. 75 FR 69030 - Airworthiness Directives; Bombardier, Inc. Model CL-600-2C10 (Regional Jet Series 700, 701, & 702...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-11-10

    .... Model CL-600-2C10 (Regional Jet Series 700, 701, & 702) Airplanes, Model CL-600-2D15 (Regional Jet Series 705) Airplanes, and Model CL-600-2D24 (Regional Jet Series 900) Airplanes AGENCY: Federal Aviation... DEPARTMENT OF TRANSPORTATION Federal Aviation Administration 14 CFR Part 39 [Docket No. FAA-2010...

  11. 76 FR 13536 - Airworthiness Directives; Bombardier, Inc. Model CL-600-2C10 (Regional Jet Series 700, 701, & 702...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-03-14

    .... Model CL-600-2C10 (Regional Jet Series 700, 701, & 702), Model CL-600-2D15 (Regional Jet Series 705), and Model CL-600-2D24 (Regional Jet Series 900) Airplanes AGENCY: Federal Aviation Administration (FAA... DEPARTMENT OF TRANSPORTATION Federal Aviation Administration 14 CFR Part 39 [Docket No. FAA-2011...

  12. Harsh Corporal Punishment Is Associated With Increased T2 Relaxation Time in Dopamine-Rich Regions

    PubMed Central

    Sheu, Yi-Shin; Polcari, Ann; Anderson, Carl M.; Teicher, Martin H.

    2010-01-01

    Harsh corporal punishment (HCP) was defined as frequent parental administration of corporal punishment (CP) for discipline, with occasional use of objects such as straps, or paddles. CP is linked to increased risk for depression and substance abuse. We examine whether long-term exposure to HCP acts as sub-traumatic stressor that contributes to brain alterations, particularly in dopaminergic pathways, which may mediate their increased vulnerability to drug and alcohol abuse. Nineteen young adults who experienced early HCP but no other forms of maltreatment and twenty-three comparable controls were studied. T2 relaxation time (T2-RT) measurements were performed with an echo planar imaging TE stepping technique and T2 maps were calculated and analyzed voxel-by-voxel to locate regional T2-RT differences between groups. Previous studies indicated that T2-RT provides an indirect index of resting cerebral blood volume. Region of interest (ROI) analyses were also conducted in caudate, putamen, nucleus accumbens, anterior cingulate cortex, dorsolateral prefrontal cortex, thalamus, globus pallidus and cerebellar hemispheres. Voxel-based relaxometry showed that HCP was associated with increased T2-RT in right caudate and putamen. ROI analyses also revealed increased T2-RT in dorsolateral prefrontal cortex, substantia nigra, thalamus and accumbens but not globus pallidus or cerebellum. There were significant associations between T2-RT measures in dopamine target regions and use of drugs and alcohol, and memory performance. Alteration in the paramagnetic or hemodynamic properties of dopaminergic cell body and projection regions were observed in subjects with HCP, and these findings may relate to their increased risk for drug and alcohol abuse. PMID:20600981

  13. Harsh corporal punishment is associated with increased T2 relaxation time in dopamine-rich regions.

    PubMed

    Sheu, Yi-Shin; Polcari, Ann; Anderson, Carl M; Teicher, Martin H

    2010-11-01

    Harsh corporal punishment (HCP) was defined as frequent parental administration of corporal punishment (CP) for discipline, with occasional use of objects such as straps, or paddles. CP is linked to increased risk for depression and substance abuse. We examine whether long-term exposure to HCP acts as sub-traumatic stressor that contributes to brain alterations, particularly in dopaminergic pathways, which may mediate their increased vulnerability to drug and alcohol abuse. Nineteen young adults who experienced early HCP but no other forms of maltreatment and twenty-three comparable controls were studied. T2 relaxation time (T2-RT) measurements were performed with an echo planar imaging TE stepping technique and T2 maps were calculated and analyzed voxel-by-voxel to locate regional T2-RT differences between groups. Previous studies indicated that T2-RT provides an indirect index of resting cerebral blood volume. Region of interest (ROI) analyses were also conducted in caudate, putamen, nucleus accumbens, anterior cingulate cortex, dorsolateral prefrontal cortex, thalamus, globus pallidus and cerebellar hemispheres. Voxel-based relaxometry showed that HCP was associated with increased T2-RT in right caudate and putamen. ROI analyses also revealed increased T2-RT in dorsolateral prefrontal cortex, substantia nigra, thalamus and accumbens but not globus pallidus or cerebellum. There were significant associations between T2-RT measures in dopamine target regions and use of drugs and alcohol, and memory performance. Alteration in the paramagnetic or hemodynamic properties of dopaminergic cell body and projection regions were observed in subjects with HCP, and these findings may relate to their increased risk for drug and alcohol abuse. Copyright 2010 Elsevier Inc. All rights reserved.

  14. Empirically constrained estimates of Alaskan regional Net Ecosystem Exchange of CO2, 2012-2014

    NASA Astrophysics Data System (ADS)

    Commane, R.; Lindaas, J.; Benmergui, J. S.; Luus, K. A.; Chang, R. Y. W.; Miller, S. M.; Henderson, J.; Karion, A.; Miller, J. B.; Sweeney, C.; Miller, C. E.; Lin, J. C.; Oechel, W. C.; Zona, D.; Euskirchen, E. S.; Iwata, H.; Ueyama, M.; Harazono, Y.; Veraverbeke, S.; Randerson, J. T.; Daube, B. C.; Pittman, J. V.; Wofsy, S. C.

    2015-12-01

    We present data-driven estimates of the regional net ecosystem exchange of CO2 across Alaska for three years (2012-2014) derived from CARVE (Carbon in the Arctic Reservoirs Vulnerability Experiment) aircraft measurements. Integrating optimized estimates of annual NEE, we find that the Alaskan region was a small sink of CO2 during 2012 and 2014, but a significant source of CO2 in 2013, even before including emissions from the large forest fire season during 2013. We investigate the drivers of this interannual variability, and the larger spring and fall emissions of CO2 in 2013. To determine the optimized fluxes, we couple the Polar Weather Research and Forecasting (PWRF) model with the Stochastic Time-Inverted Lagrangian Transport (STILT) model, to produce footprints of surface influence that we convolve with a remote-sensing driven model of NEE across Alaska, the Polar Vegetation Photosynthesis and Respiration Model (Polar-VPRM). For each month we calculate a spatially explicit additive flux (ΔF) by minimizing the difference between the measured profiles of the aircraft CO2 data and the modeled profiles, using a framework that combines a uniform correction at regional scales and a Bayesian inversion of residuals at smaller scales. A rigorous estimate of total uncertainty (including atmospheric transport, measurement error, etc.) was made with a combination of maximum likelihood estimation and Monte Carlo error propagation. Our optimized fluxes are consistent with other measurements on multiple spatial scales, including CO2 mixing ratios from the CARVE Tower near Fairbanks and eddy covariance flux towers in both boreal and tundra ecosystems across Alaska. For times outside the aircraft observations (Dec-April) we use the un-optimized polar-VPRM, which has shown good agreement with both tall towers and eddy flux data outside the growing season. This approach allows us to robustly estimate the annual CO2 budget for Alaska and investigate the drivers of both the

  15. Differential requirements for Gli2 and Gli3 in the regional specification of the mouse hypothalamus

    PubMed Central

    Haddad-Tóvolli, Roberta; Paul, Fabian A.; Zhang, Yuanfeng; Zhou, Xunlei; Theil, Thomas; Puelles, Luis; Blaess, Sandra; Alvarez-Bolado, Gonzalo

    2015-01-01

    Secreted protein Sonic hedgehog (Shh) ventralizes the neural tube by modulating the crucial balance between activating and repressing functions (GliA, GliR) of transcription factors Gli2 and Gli3. This balance—the Shh-Gli code—is species- and context-dependent and has been elucidated for the mouse spinal cord. The hypothalamus, a forebrain region regulating vital functions like homeostasis and hormone secretion, shows dynamic and intricate Shh expression as well as complex regional differentiation. Here we asked if particular combinations of Gli2 and Gli3 and of GliA and GliR functions contribute to the variety of hypothalamic regions, i.e., we wanted to approach the question of a possible hypothalamic version of the Shh-Gli code. Based on mouse mutant analysis, we show that: (1) hypothalamic regional heterogeneity is based in part on differentially stringent requirements for Gli2 or Gli3; (2) another source of diversity are differential requirements for Shh of neural vs. non-neural origin; (3) the medial progenitor domain known to depend on Gli2 for its development generates several essential hypothalamic nuclei plus the pituitary and median eminence; (4) the suppression of Gli3R by neural and non-neural Shh is essential for hypothalamic specification. Finally, we have mapped our results on a recent model which considers the hypothalamus as a transverse region with alar and basal portions. Our data confirm the model and are explained by it. PMID:25859185

  16. [Variation of insulin receptor substrate-2 gene 3'-untranslated region in patients with type 2 diabetes mellitus].

    PubMed

    Zeng, Wei-Min; Chen, Shu-Hua; Xie, Ping; Liu, Mei-Lian; Song, Hui-Ping

    2003-08-01

    Insulin receptor substrate-2(IRS-2) belongs to a family of cytoplasmic adaptor proteins, which link insulin, insulin-like growth factor-1(IGF-1), and cytokine receptor tyrosine kinases to signaling pathways regulating metabolism, growth, differentiation, reproduction, and homestasis. Deficiency of IRS-2 in mice causes type 2 diabetes mellitus (T2DM), suggesting that abnormal structure and dysfunction of the IRS-2 gene may contribute to the pathogenesis of T2DM. Variations in the open reading frame (ORF) and promoter region of IRS-2 gene in patients with T2DM have been reported over the past few years. These genetic variations are from ethnically different patients, confounding any analysis of the contribution of IRS-2 gene variations to the development of T2DM. The 3'-untranslated region(3'-UTR) of IRS-2 gene variation may be contribute to the T2DM. So far, the relationship between 3'-UTR of IRS-2 gene variations and T2DM have not been investigated. Based on the 3'-UTR of eukaryotic gene plays an important role in the eukaryotic gene regulation, we investigated abnormalities of IRS-2 gene 3'-UTR and their relation with T2DM in the Chinese population. Genomic DNA was extracted from leukocyte of 128 patients with T2DM and 125 control subjects in Hunan, China. A segment of IRS-2 gene 3'-UTR was scanned by polymerase chain reaction (PCR)-denaturing high-performance liquid chromatography (DHPLC). All PCR products with abnormal DHPLC pattern were submitted to DNA sequence analysis. A T-->C mutation at 4064 bp of IRS-2 gene 3'-UTR was found in 18 patients with T2DM, while it was only found in 5 control subjects. The incidence of the mutation in patients with T2DM were much higher than that in contol subjects (14.1% vs 4.0%, x2 = 7.748, P = 0.005). These results indicate that the T4064-->C in IRS-2 gene 3'-UTR may be related to Chinese patients with T2DM.

  17. Umgang mit Antithrombotika bei Operationen an der Haut vor und nach Publikation der entsprechenden S3-Leitlinie.

    PubMed

    Gaskins, Matthew; Dittmann, Martin; Eisert, Lisa; Werner, Ricardo Niklas; Dressler, Corinna; Löser, Christoph; Nast, Alexander

    2018-03-01

    Laut einer Befragung im Jahre 2012 war der Umgang mit Antithrombotika bei dermatochirurgischen Eingriffen in Deutschland sehr heterogen. 2014 wurde erstmals eine evidenzbasierte Leitlinie zu diesem Thema veröffentlicht. Es wurde eine anonyme Befragung derselben Stichprobe zum Umgang mit Antithrombotika sowie zu Kenntnissen der Leitlinie durchgeführt. Die Ergebnisse wurden als relative Häufigkeiten berichtet und denen aus 2012 gegenübergestellt. 208 Antwortbögen wurden ausgewertet (Rücklaufquote: 36,6 %). Die große Mehrheit der Dermatologen erklärte, kleinere Eingriffe unter Fortführung der Therapie mit Phenprocoumon, niedrig dosierter Acetylsalicylsäure (≤ 100 mg) und Clopidogrel sowie mit direkten oralen Antikoagulanzien durchzuführen. Bei größeren Eingriffen war der Umgang hingegen weiterhin heterogen, insbesondere unter niedergelassenen Dermatologen. Der Anteil der Dermatologen, die Phenprocoumon, Acetylsalicylsäure und Clopidogrel leitlinienkonform verwendeten, hat sich insgesamt vergrößert. Führten 2012 beispielsweise 53,8 % der Klinikärzte bzw. 36,3 % der niedergelassenen Dermatologen eine große Exzision unter Fortführung der Therapie mit niedrig dosierter Acetylsalicylsäure durch, taten dies 2017 90,2 % bzw. 57,8 % (Phenprocoumon: 33,8 % bzw. 11,9 % auf 63,9 % bzw. 29,9 %; Clopidogrel: 36,9 % bzw. 23,2 % auf 63,9 % bzw. 30,6 %). Unter den Klinikärzten war ein hoher Anteil mit der Leitlinie vertraut und fand diese hilfreich. Eine Zunahme des leitlinienkonformen Verhaltens war bei allen Eingriffen zu verzeichnen. Bei größeren Eingriffen zeigte sich trotz deutlicher Verbesserung die Notwendigkeit verstärkter Anstrengungen zur Leitlinienumsetzung bzw. zur Identifizierung von Implementierungsbarrieren. © 2018 Deutsche Dermatologische Gesellschaft (DDG). Published by John Wiley & Sons Ltd.

  18. Irradiated interfaces in the Ara OB1, Carina, Eagle Nebula, and Cyg OB2 massive star formation regions

    DOE PAGES

    Hartigan, P.; Palmer, J.; Cleeves, L. I.

    2012-09-05

    Regions of massive star formation offer some of the best and most easily-observed examples of radiation hydrodynamics. Boundaries where fully-ionized H II regions transition to neutral/molecular photodissociation regions (PDRs) are of particular interest because marked temperature and density contrasts across the boundaries lead to evaporative flows and fluid dynamical instabilities that can evolve into spectacular pillar-like structures. Furthermore, when detached from their parent clouds, pillars become ionized globules that often harbor one or more young stars. H2 molecules at the interface between a PDR and an H II region absorb ultraviolet light from massive stars, and the resulting fluoresced infraredmore » emission lines are an ideal way to trace this boundary independent of obscuring dust. This paper presents H2 images of four regions of massive star formation that illustrate different types of PDR boundaries. The Ara OB1 star formation region contains a striking long wall that has several wavy structures which are present in H2, but the emission is not particularly bright because the ambient UV fluxes are relatively low. In contrast, the Carina star formation region shows strong H2 fluorescence both along curved walls and at the edges of spectacular pillars that in some cases have become detached from their parent clouds. The less-spectacular but more well-known Eagle Nebula has two regions that have strong fluorescence in addition to its pillars. And while somewhat older than the other regions, Cyg OB2 has the highest number of massive stars of the regions surveyed and contains many isolated, fluoresced globules that have head–tail morphologies which point towards the sources of ionizing radiation. Our images provide a collection of potential astrophysical analogs that may relate to ablated interfaces observed in laser experiments of radiation hydrodynamics.« less

  19. H II REGIONS, EMBEDDED PROTOSTARS, AND STARLESS CORES IN SHARPLESS 2-157

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Chen, Chian-Chou; Williams, Jonathan P.; Pandian, Jagadheep D., E-mail: ccchen@ifa.hawaii.edu, E-mail: jpw@ifa.hawaii.edu, E-mail: jagadheep@iist.ac.in

    2012-06-20

    We present arcsecond resolution 1.4 mm observations of the high-mass star-forming region, Sharpless 2-157, that reveal the cool dust associated with the first stages of star formation. These data are compared with archival images at optical, infrared, and radio wavelengths, and complemented with new arcsecond resolution mid-infrared data. We identify a dusty young H II region, numerous infrared sources within the cluster envelope, and four starless condensations. Three of the cores lie in a line to the south of the cluster peak, but the most massive one is right at the center and associated with a jumble of bright radiomore » and infrared sources. This presents an interesting juxtaposition of high- and low-mass star formation within the same cluster which we compare with similar observations of other high-mass star-forming regions and discuss in the context of cluster formation theory.« less

  20. Very Large Array H I Zeeman Observations of the Cygnus X Region: DR 22 and ON 2

    NASA Astrophysics Data System (ADS)

    Mayo, E. A.; Troland, T. H.

    2012-02-01

    We have used the Very Large Array to study the Zeeman effect in 21 cm H I absorption lines from two star-forming regions in the Cygnus X complex, DR 22 and ON 2. We measure the line-of-sight magnetic field toward these regions, finding B los = -84 ± 11 μG toward the DR 22 H II region and B los < 50 μG toward each of the two H II regions in ON 2. We interpret these results in terms of two different models. In one model, we assume that the H I Zeeman effect is a measure of magnetic fields in the associated molecular clouds. If so, then the DR 22 molecular cloud is magnetically subcritical, that is, magnetically dominated. The ON 2 molecular clouds are magnetically supercritical. In a second model, we assume that the H I Zeeman effect is a measure of magnetic fields in photon-dominated regions where the gas has been compressed (and the field amplified) by absorption of stellar radiation. We find that this second model, where the measured field strength has been affected by star formation, accounts well for the DR 22 H I Zeeman effect. This same model, however, overpredicts the magnetic field in ON 2. ON 2 may be a region where the magnetic field is energetically insignificant or where the field happens to lie nearly in the plane of the sky.