Disaggregated N2O emission factors in China based on cropping parameters create a robust approach to the IPCC Tier 2 methodology

Acknowledgements This work was funded by Chinese Ministry of Agriculture and the United Kingdom Department for Environment, Food and Rural Affairs (DEFRA), UK under the UK-China Sustainable Agriculture Innovation Network (SAIN; Project DC09-06). Rothamsted Research receives strategic funding by the Biotechnology and Biological Sciences Research Council (BBSRC).Peer reviewedPublisher PD

Frequency and Distribution of Pseudomonas aeruginosa Serotypes 03, 06, 011 in Three Northwestern Ohio Hospitals as Determined by ELISA Using Specific Monoclonal Antibodies

Author Institution: Medical Technology and Biological Sciences, Bowling Green State UniversityHybridoma producing monoclonal antibodies (mAbs), specific for three clinically significant Pseudomonas aeruginosa (serotypes 03, 06, and Oil), were generated to investigate the prevalence of these serotypes in three Northwestern Ohio hospitals. Fusion products reacting with bacterial cells or membrane extracts were detected by enzyme-linked-immunosorbent assay (ELISA). Three mAbs designated: 72C, 11 H (IgM) and HE (IgG2b), were selected. These mAbs reacted with approximately 40% of the clinical isolates of P. aeruginosa in each hospital. The incidence of serotype Oil varied in these hospitals, ranging from 13.2%-23.8%. Serotype Oil predominated in two of the three hospitals. The prevalence of serotype 06 was similar in all three hospitals (13.6-15%). In one of the hospitals (Hospital 2), the occurrence of serotype 06 was slightly higher (15%) than serotype Oil (13-2%). Serotype 03 occurred less frequently (1.5%) in one of the hospitals than in the other two (10-11%). None of the serotypes showed clear predilection toward any body site. The mAbs did not react with other strains of P. aeruginosa, nor with other gram-negative or gram-positive organisms. The results of Immunofluorescence and Western blot correlated well with ELISA. However, ELISA showed a higher sensitivity, indicating the usefulness of this technique for serotyping P. aeruginosa

Substrate Fragmentation for the Design of M. tuberculosis CYP121 Inhibitors.

The cyclo-dipeptide substrates of the essential M. tuberculosis (Mtb) enzyme CYP121 were deconstructed into their component fragments and screened against the enzyme. A number of hits were identified, one of which exhibited an unexpected inhibitor-like binding mode. The inhibitory pharmacophore was elucidated, and fragment binding affinity was rapidly improved by synthetic elaboration guided by the structures of CYP121 substrates. The resulting inhibitors have low micromolar affinity, good predicted physicochemical properties and selectivity for CYP121 over other Mtb P450s. Spectroscopic characterisation of the inhibitors' binding mode provides insight into the effect of weak nitrogen-donor ligands on the P450 heme, an improved understanding of factors governing CYP121-ligand recognition and speculation into the biological role of the enzyme for Mtb.M.E.K. was supported by a Commonwealth (University of Cambridge) Scholarship awarded in conjunction with the Cambridge Commonwealth Trust and Cambridge Overseas Trust. A.G.C. and K.J.M. were supported by grants from the Biotechnology and Biological Sciences Research Council (BBSRC) (grant nos. BB/I019669/1 and BB/I019227/1). We acknowledge the support of the Wellcome Trust (Translation Award GR080083/Z/06).This is the final version of the article. It first appeared from Wiley at http://dx.doi.org/10.1002/cmdc.201600248

Characterization of Ribosomal Frameshifting in Theiler's Murine Encephalomyelitis Virus.

Theiler's murine encephalomyelitis virus (TMEV) is a member of the genus Cardiovirus in the Picornaviridae, a family of positive-sense single-stranded RNA viruses. Previously, we demonstrated that in the related cardiovirus, Encephalomyocarditis virus, a programmed-1 ribosomal frameshift (1 PRF) occurs at a conserved G_GUU_UUU sequence within the 2B-encoding region of the polyprotein open reading frame (ORF). Here we show that-1 PRF occurs at a similar site during translation of the TMEV genome. In addition, we demonstrate that a predicted 3= RNA stem-loop structure at a noncanonical spacing downstream of the shift site is required for efficient frameshifting in TMEV and that frameshifting also requires virus infection. Mutating the G_GUU_UUU shift site to inhibit frameshifting results in an attenuated virus with reduced growth kinetics and a small-plaque phenotype. Frameshifting in the virus context was found to be extremely efficient at 74 to 82%, which, to our knowledge, is the highest frameshifting efficiency recorded to date for any virus. We propose that highly efficient-1 PRF in TMEV provides a mechanism to escape the confines of equimolar expression normally inherent in the single-polyprotein expression strategy of picornaviruses.Work in the A.E.F. lab is supported by the Wellcome Trust [088789], [106207]; and the Biotechnology and Biological Sciences Research Council [BB/J007072/1]. L.F. is supported by a Biotechnology and Biological Sciences Research Council PhD studentship.This is the final published version. It first appeared at http://jvi.highwire.org/content/early/2015/06/05/JVI.01043-15.abstract

Protease inhibitors extracted from caesalpinia echinata lam. Affect kinin release during lung inflammation

Inflammation is an essential process in many pulmonary diseases in which kinins are generated by protease action on kininogen, a phenomenon that is blocked by protease inhibitors. We evaluated kinin release in an in vivo lung inflammation model in rats, in the presence or absence of CeKI (C. echinata kallikrein inhibitor), a plasma kallikrein, cathepsin G, and proteinase-3 inhibitor, and rCeEI (recombinant C. echinata elastase inhibitor), which inhibits these proteases and also neutrophil elastase. Wistar rats were intravenously treated with buffer (negative control) or inhibitors and, subsequently, lipopolysaccharide was injected into their lungs. Blood, bronchoalveolar lavage fluid (BALF), and lung tissue were collected. In plasma, kinin release was higher in the LPS-treated animals in comparison to CeKI or rCeEI groups. rCeEI-treated animals presented less kinin than CeKI-treated group. Our data suggest that kinins play a pivotal role in lung inflammation and may be generated by different enzymeshowever, neutrophil elastase seems to be the most important in the lung tissue context. These results open perspectives for a better understanding of biological process where neutrophil enzymes participate and indicate these plant inhibitors and their recombinant correlates for therapeutic trials involving pulmonary diseases.Fundacao de Amparo a Pesquisa do Estado de Sao Paulo [04/11015-0, 07/55496-0, 01/02457-0]Conselho Nacional de Desenvolvimento Cientifico e Tecnologico [304923/2006-0, 304719/2009-9]Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior/Ministerio da Educacao Superior de Cuba (CAPES/MES), Brazil [011/06, 077/09]Department of Biochemistry, Universidade Federal de S˜ao Paulo, Rua Trˆes de Maio, No. 100, 04044-020 S˜ao Paulo, SP, BrazilSchool of Arts, Sciences and Humanities, Universidade de São Paulo, Avenida Arlindo Bettio, No. 1000, 03828-000 São Paulo, SP, BrazilDepartment of Marine Sciences, Universidade Federal de São Paulo, Rua Doutor Carvalho de Mendonça, No. 144, 11070-100 Santos, SP, BrazilJapan Health Care College, Sinei 434-1, Kiyota-ku, Sapporo, JapanDepartment of Marine Sciences, Universidade Federal de São Paulo, Rua Doutor Carvalho de Mendonça, No. 144, 11070-100 Santos, SP, BrazilFAPESP: 04/11015-0FAPESP: 07/55496-0FAPESP: 01/02457-0CNPq: 304923/2006-0CNPq: 304719/2009-9CAPES/MES: 011/06 and 077/09Web of Scienc

Selective Inhibition of Heparan Sulphate and Not Chondroitin Sulphate Biosynthesis by a Small, Soluble Competitive Inhibitor

From MDPI via Jisc Publications RouterHistory: accepted 2021-06-19, pub-electronic 2021-06-29Publication status: PublishedFunder: Biotechnology and Biological Sciences Research Council; Grant(s): 978724Funder: Medical Research Council; Grant(s): MR/L007525/1The glycosaminoglycan, heparan sulphate (HS), orchestrates many developmental processes. Yet its biological role has not yet fully been elucidated. Small molecule chemical inhibitors can be used to perturb HS function and these compounds provide cheap alternatives to genetic manipulation methods. However, existing chemical inhibition methods for HS also interfere with chondroitin sulphate (CS), complicating data interpretation of HS function. Herein, a simple method for the selective inhibition of HS biosynthesis is described. Using endogenous metabolic sugar pathways, Ac4GalNAz produces UDP-GlcNAz, which can target HS synthesis. Cell treatment with Ac4GalNAz resulted in defective chain elongation of the polymer and decreased HS expression. Conversely, no adverse effect on CS production was observed. The inhibition was transient and dose-dependent, affording rescue of HS expression after removal of the unnatural azido sugar. The utility of inhibition is demonstrated in cell culture and in whole organisms, demonstrating that this small molecule can be used as a tool for HS inhibition in biological systems

Fossil biomass preserved as graphitic carbon in a late paleoproterozoic banded iron formation metamorphosed at more than 550°C

Metamorphism is thought to destroy microfossils, partly through devolatilization and graphitization of biogenic organic matter. However, the extent to which there is a loss of molecular, elemental and isotope signatures from biomass during high-temperature metamorphism is not clearly established. We report on graphitic structures inside and coating apatite grains from the c. 1850 Ma Michigamme silicate banded iron formation from Michigan, metamorphosed above 550°C. Traces of N, S, O, H, Ca and Fe are preserved in this graphitic carbon and X-ray spectra show traces of aliphatic groups. Graphitic carbon has an expanded lattice around 3.6 Å, forms microscopic concentrically-layered and radiating polygonal flakes and has homogeneous δ13C values around −22‰, identical to bulk analyses. Graphitic carbon inside apatite is associated with nanometre-size ammoniated phyllosilicate. Precursors of these metamorphic minerals and graphitic carbon originated from ferruginous clayrich sediments with biomass. We conclude that graphite coatings and inclusions in apatite grains indicate fluid remobilization during amphibolite-facies metamorphism of precursor biomass. This new evidence fills in observational gaps of metamorphosed biomass into graphite and supports the existence of biosignatures in the highly metamorphosed iron formation from the Eoarchean Akilia Association, which dates from the beginning of the sedimentary rock record

Evidence-based Decision-making in Canada’s Protected Areas Organizations: Implications for Management Effectiveness

Aichi Biodiversity Target 19 calls on Parties to the United Nations Convention on Biological Diversity (CBD) to improve, share, transfer, and apply knowledge. In this study, we provide an initial assessment of the state of evidence-based decision-making in Canada’s protected areas organizations by examining (1) the value and use of various forms of evidence by managers and (2) the extent to which institutional conditions enable or inhibit the use of evidence in decision-making. Results revealed that although managers value and use many forms of evidence in their decision-making, information produced by staff and their organizations are given priority. Other forms of evidence, such as Indigenous knowledge and peer-reviewed information, are valued and used less. The most significant barriers to evidence-based decision-making were limited financial resources, lack of staff, inadequate timeframes for decision-making, a lack of monitoring programs, and a disconnect between researchers and decision-makers. Overall, our results suggest that the potential benefits of evidence-based approaches are not being maximized in Canada’s protected areas organizations. We propose several recommendations to introduce or improve the use of diverse forms of evidence to enhance management effectiveness of Canada’s protected areas and by extension conservation outcomes