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Plasmid instability when the hsp60 gene promoter is used to express the protective non-toxic fragment b of the diphtheria toxin in recombinant bCG

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American Journal of molecular biology 2013年第2期3卷 81-86页

作者： Dilzamar V.Nascimento Odir A.Dellagostin Raphael Hirata Jr geraldo m.b.pereira Ana Luiza mattos-Guaraldi geraldo R.Garcia Armoa Faculdade de Ciencias médicas Universidade do Estado do Rio de JaneiroRio de JaneiroBrazil Centro de biotecnologia Universidade Federal de PelotasCampus UniversitárioPelotasBrazil Instituto Oswaldo Cruz Fundacao Oswaldo CruzRio de JaneiroBrazil

The genetic modification of the live attenuated mycobacterium bovis bCG to deliver a protective Corynebacterium diphtheriae antigen in vivo could be a safer and less costly alternative to the new and more expensive DTP vaccines available today, in particular to third world-countries. The stability of expression of heterologous antigens in bCG, however, is a major challenge to the use of live recombinant bacteria in vaccine development and appears to be dependent to a certain extent, on a genetic compatibility between the expression cassette within the plasmid construct and the mycobacterium host. In the quest for the best recombinant bCG transformant to express the dtb gene of C. diphtheriae we generated two new rbCG strains by transforming the moreau substrain of bCG with the mycobacterial expression vectors pUS973 and pUS977, each one carrying a different promoter to drive the expression of the target antigen. After transformation recombinant bCG clones were selected on middlebrook 7H10 kanamycin Agar plates, expanded in middlebrook 7H9 kanamycin broth and analyzed by agarose gel electrophoresis and immunoblotting. rbCGs transformed with the construct carrying the weak PAN promoter from m. paratuberculosis stably expressed the dtb gene. Conversely, rbCGs transformed with the construct carrying the strong mycobacterium hsp60 promoter were unstable and consequently unfit for the expression of the C. diphtheriae gene.

关键词： bCG Vaccine Recombinant bCG Fragment b of Diphtheria Toxin Anti-Diphtheria Vaccine

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Carbon monoxide as C1 building block in Fine Chemical Synthesis

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Chinese Journal of Chemistry 2024年第2期42卷 199-221页

作者： Rui m.b.Carrilho mario J.F.Calvete Gabor mikle Laszlo Kollar mariette m.pereira University of Coimbra Coimbra Chemistry CentreDepartment of ChemistryRua Larga3004-535 CoimbraPortugal Department of General and Inorganic Chemistry and ELKH-PTE Research Groupfor Selective Chemical Syntheses University of PecsIfjusag utja 6H-7624 PecsHungary

Carbon monoxide(CO)has become one of the most relevant and versatile renewable C1 building blocks for chemical synthesis,especially in the fine chemicals industry,due to the development of efficient and selective catalysts for its *** this review,we present a comprehensive critical analysis of the last 10 years literature on the use of CO as a renewable feedstock for fine chemicals *** review is organized by type of catalytic reaction,namely alkene and alkyne carbonylation,hydroformylation,carbonylation of aryl halides,carbonylative cross-coupling and C-H *** examples of the synthesis of relevant building blocks and/or known pharmaceuticals are *** is placed on examples of utilizing CO as the C1 building block in one or more catalytic *** catalyst used and the reaction conditions are consistently presented throughout all of the examples.

关键词： Carbon monoxide Fine chemicals Carbonylation Hydroformylation C-H functionalization

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mycobacterium bovis bCG as a Delivery System for the dtb Gene Antigen from Diphtheria Toxin

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American Journal of molecular biology 2017年第4期7卷 176-189页

作者： Dilzamar V.Nascimento Odir A.Dellagostin Denise C.S.matos Douglas mcIntosh Raphael Hirata Jr. geraldo m.b.pereira Ana Luíza mattos-Guaraldi geraldo R.G.Armoa Faculdade de Ciências médicas Universidade do Estado do Rio de JaneiroRio de JaneiroBrazil Núcleo de biotecnologia Universidade Federal de PelotasCampus UniversitárioPelotasBrazil Instituto de Tecnologia em Imunobiológicos Fundacao Oswaldo CruzRio de JaneiroBrazil Instituto Oswaldo Cruz Fundacao Oswaldo CruzRio de JaneiroBrazil Instituto de Veterinária Universidade Federal Rural do Rio de JaneiroSeropedicaRio de JaneiroBrazil

Diphtheria is a fulminant bacterial disease caused by toxigenic strains of Corynebacterium diphtheriae whose local and systemic manifestations are due to the action of the diphtheria toxin (DT). The vaccine which is used to prevent diphtheria worldwide is a toxoid obtained by detoxifying DT. Although associated with high efficacy in the prevention of disease, the current anti-diphtheria vaccine, one of the components of DTP (diphtheria, tetanus and pertussis triple vaccine), may present post vaccination effects such as toxicity and reactogenicity resulting from the presence of contaminants in the vaccine that originated during the process of production and/or detoxification. Therefore, strategies to develop a less toxic and at the same time economically viable vaccine alternatives are needed to improve existing vaccines in use worldwide. In this study, the moreau substrain of bCG which is used in brazil as a live vaccine against human tuberculosis was genetically modified to carry and express the gene encoding for the diphtheria toxin fragment b (DTb). As such, the DNA sequence encoding the dtb gene was cloned into the pUS977 shuttle vector for cytoplasmic expression and successfully introduced into bCG cells by electroporation. mice immunized with recombinant bCG expressing DTb showed seroconversion with the detection of specific antibodies against DTb. Also, rbCGs stably expressing DTb persisted up to 60 days in the absence of selective pressure in mice and cell viability did not change significantly during the period tested. Finally, immune sera from bALb/c mice vaccinated with rbCGpUS977dtbPW8 were preliminarily tested for their capacity of neutralizing the diphtheria toxin in the Vero Cells assay.

关键词： Recombinant bCG Diphtheria Toxin dtb Gene Park Williams 8(PW8) Corynebacterium diphtheriae rDTbPW8 pUS977 Vector

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A geomorphological model of susceptibility to the effect of human interventions for environmental licensing determination(SHIELD)

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Geoscience Frontiers 2022年第2期13卷 284-294页

作者： Cristina I.pereira Celene b.milanes Ivan Correa Enzo Pranzini benjamin Cuker Camilo m.botero EAFIT University Earth Sciences DepartmentMedellínColombia Coastal Systems Research Group Playas Corporation Ltda.BogotáColombia Civil and Environmental Department Universidad De La CostaBarranquillaColombia Department of Earth Sciences University of FlorenceFirenzeItaly Department of marine and Environmental Science Hampton UniversityHamptonUnited States University Sergio Arboleda School of LawSanta

Almost every country requires some form of environmental licensing prior to the inception of development projects that may affect the integrity of the environment and its social *** developed a new conceptual and methodological model to instruct the assessment of the potential impacts posed by proposed *** to Human Interventions for Environmental Licensing Determination(SHIELD)includes a novel geomorphological interpretation of the Environmental Impact Assessment(EIA).It considers the impact of human interventions on geomorphological processes and landscape functioning in the context of the entire ecosystem,going further than the classical concept of *** susceptibility of the site informs the screening stage,allowing local conditions to help define the criteria used in the ***,the level of detail of the environmental baseline is scoped by considering the degree of disturbance of natural processes posed by human *** this geomorphological susceptibility model on different kinds of environments would allow shifting the environmental licensing practices from the prevailing anthropocentric and static conception of the environment towards an Ecosystem *** addresses the need to improve the screening and scoping stages that form the basis of the rest of any *** introduces several innovations to EIA including the incorporation of fuzzy logic,a preassembled database of contributions form experts,and a shifting of emphasis from the type of proposed intervention to the type of environment and its relative susceptibility.

关键词： Processes and landforms Expert knowledge Ecosystem approach Screening Scoping

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Large ferro-pyro-phototronic effect in 0.5ba(Zr_(0.2)Ti_(0.8))O_(3)-0.5(ba_(0.7)Ca_(0.3))TiO_(3)thin films integrated on silicon for photodetection

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Carbon Energy 2023年第6期5卷 1-10页

作者： JoséP.b.Silva Katarzyna Gwozdz Luís S.marques mario pereira maria J.m.Gomes Judith L.macmanus‐Driscoll Robert L.Z.Hoye Centre of Physics of Universities of minho and Porto(CF‐Um‐UP) University of MinhoBragaPortugal Laboratory of Physics for materials and Emergent Technologies LapMETUniversity of MinhoBragaPortugal Department of Quantum Technologies Faculty of Fundamental Problems of TechnologyWroclaw University of Science and TechnologyWroclawPoland Department of materials Science and metallurgy CambridgeUK Inorganic Chemistry Laboratory Department of ChemistryUniversity of OxfordOxfordUK Department of materials Imperial College LondonLondonUK

Coupling together the ferroelectric,pyroelectric,and photovoltaic characteristics within a single material is a novel way to improve the performance of *** this work,we take advantage of the triple multifunctionality shown by 0.5ba(Zr_(0.2)Ti_(0.8))O_(3)-0.5(ba_(0.7)Ca_(0.3))TiO_(3)(bCZT),as demonstrated in an Al/Si/SiOx/bCZT/ITO thin-film *** Si/SiOx acts as an n-type layer to form a metal-ferroelectric-insulator-semiconductor heterostructure with the bCZT,and with Al and ITO as *** photo-response of the device,with excitation from a violet laser(405 nm wavelength),is carefully investigated,and it is shown that the photodetector performance is invariant with the chopper frequency owing to the pyro-phototronic effect,which corresponds to the coupling together of the pyroelectric and photovoltaic ***,the photodetector performance was significantly better than that of the devices operating based only on the pyro-phototronic effect by a factor of 4,due to the presence of ferroelectricity in the ***,after a poling voltage of−15 V,for a laser power density of 230mW/cm^(2)and at a chopper frequency of 400Hz,optimized responsivity,detectivity,and sensitivity values of 13.1mA/W,1.7×10^(10)Jones,and 26.9,respectively,are ***,ultrafast rise and fall times of 2.4 and 1.5μs,respectively,are obtained,which are 35,000 and 36,000 times faster rise and fall responses,respectively,than previous reports of devices with the ferro-pyro-phototronic *** is understood based on the much faster ferroelectric switching in ferroelectric thin films owing to the predominant 180°domains in a single direction out of plane.

关键词： ferroelectric ferro-pyro-phototronic effect photodetector polarization-controlled response

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Assessing tree crown fire damage integrating linear spectral mixture analysis and supervised machine learning on Sentinel-2 imagery

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International Journal of Digital Earth 2023年第1期16卷 3162-3198页

作者： Giandomenico De Luca Giuseppe modica Joao m.N.Silva Salvatore Praticò Josém.C.pereira Dipartimento di Agrari1 Universitàdegli Studi Mediterranea di Reggio Calabri1Reggio Calabri1Italy Dipartimento di Scienze veterinarie Universitàdegli Studi di Messin1Messin1Italy Forest Research Centre School of AgricultureUniversity of LisbonLisbonPortugal

Crown fire damage is a mixture of three principal fire-related components:charred material,scorched foliage,and unaltered green *** study estimated the abundance of these physical alterations in two immediate post-fire mediterranean forest contexts(Portugal and Italy)by applying linear spectral mixture analysis(LSmA)on Sentinel-2 *** tree crowns fire damage was subsequently mapped,integrating fractional abundance information in a random forest(RF)algorithm,comparing the accuracy resulting from the adoption of generic or image spectral libraries as the primary investigative *** image-derived endmembers resulted in more effectiveness in terms of fire-related components abundance quantification(LmSAderived RmSEmembers or image endmembers derived information was *** environmental heterogeneity of the two study areas affected the fire severity gradients,with a prevalence of the charred(PT)(45–46%)and green class(IT)(44–53%).Post-fire temporal monitoring was initialized by applying the proposed strategies,and the preliminary results showed a positive recovery trend in forest vegetation from the first year following the fire event,with a reduced charcoal predominance and an increasing proportion of green components.

关键词： Post-fire assessment fire severity post-fire vegetation recovery random forest(RF) scikit-learn fraction image extraction spectral unmixing endmembers crown fire damage mapping fully constrained least squares pixel purity index

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Challenges at the early stages of the environmental licensing procedure and potential contributions from geomorphology

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Geoscience Frontiers 2021年第6期12卷 36-45页

作者： Cristina I.pereira Celene b.milanes Rafael Sarda benjamin Cuker Camilo m.botero Coastal Systems Research Group Playas Corporation Ltda.BogotáColombia Corporación para el Desarrollo Sostenible del Urabá–CORPOURAbA Barrio ManzanaresCl.92No 98-39ApartadóColombia Civil and Environmental Department Universidad de la CostaBarranquillaColombia Centro de Estudios Avanzados de blanes Consejo Superior de Investigaciones CientíficasCEAB-CSICSpain Department of marine and Environmental Science Hampton UniversityHamptonVA 23668USA University Sergio Arboleda School of LawSanta MartaColombia

Defining impact significance is the main technical task that influences decision-making during the Environmental Licensing Procedure(ELP).The ELP begins with screening to determine potentially significant impacts of the proposed *** then follows to address any interventions deemed worthy of attention in the production of an Environmental Impact Assessment(EIA).This will include consideration of relevant landforms and geomorphological ***,preliminary assessments of environmental impacts often lack the scientific robustness to procure substantive and transactive *** review presents an examination of the established practices of screening and scoping while highlighting the foremost challenges to improve the technical grounds of the *** analysis of screening and scoping practices stresses the need for novel methods that ensure the sequential reasoning between their criteria while improving the preliminary evaluation of impact *** the inherent subjectivity of discretionary judgment requires scientific methodologies that acknowledge the interaction between the natural system and human interventions,which has been addressed by geomorphological *** knowledge consolidated in this review opens the gate to explore the compatibility between the United Nations strategy of Ecosystem Approach(EA)with the ELP through a novel geomorphological interpretation of the ***,this diagnosis demonstrate that screening and scoping practices would benefit from reliable methods that balance the precautionary principle with the efficient character required in the ELP.

关键词： EIA effectiveness Screening and scoping practices Landforms Geomorphological processes Ecosystem approach

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Performance and calibration of quark/gluon-jet taggers using 140 fb^(-1) of pp collisions at √s=13 TeV with the ATLAS detector

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Chinese Physics C 2024年第2期48卷 1-25页

作者： G. Aad b. Abbott K. Abeling N.J. Abicht S.H. Abidi A. Aboulhorma H. Abramowicz H. Abreu Y. Abulaiti b.S. Acharya C. Adam bourdarios L. Adamczyk S.V. Addepalli m.J. Addison J. Adelman A. Adiguzel T. Adye A.A. Affolder Y. Afik m.N. Agaras J. Agarwala A. Aggarwal C. Agheorghiesei A. Ahmad F. Ahmadov W.S. Ahmed S. Ahuja X. Ai G. Aielli A. Aikot m. Ait Tamlihat b. Aitbenchikh I. Aizenberg m. Akbiyik T.P.A. Åkesson A.V. Akimov D. Akiyama N.N. Akolkar K. Al Khoury G.L. Alberghi J. Albert P. Albicocco G.L. Albouy S. Alderweireldt m. Aleksa I.N. Aleksandrov C. Alexa T. Alexopoulos F. Alfonsi m. Algren m. Alhroob b. Ali H.m.J. Ali S. Ali S.W. Alibocus m. Aliev G. Alimonti W. Alkakhi C. Allaire b.m.m. Allbrooke J.F. Allen C.A. Allendes Flores P.P. Allport A. Aloisio F. Alonso C. Alpigiani m. Alvarez Estevez A. Alvarez Fernandez m. Alves Cardoso m.G. Alviggi m. Aly Y. Amaral Coutinho A. Ambler C. Amelung m. Amerl C.G. Ames D. Amidei S.P. Amor Dos Santos K.R. Amos V. Ananiev C. Anastopoulos T. Andeen J.K. Anders S.Y. Andrean A. Andreazza S. Angelidakis A. Angerami A.V. Anisenkov A. Annovi C. Antel m.T. Anthony E. Antipov m. Antonelli F. Anulli m. Aoki T. Aoki J.A. Aparisi Pozo m.A. Aparo L. Aperio bella C. Appelt A. Apyan N. Aranzabal S.J. Arbiol Val C. Arcangeletti A.T.H. Arce E. Arena J-F. Arguin S. Argyropoulos J.-H. Arling O. Arnaez H. Arnold G. Artoni H. Asada K. Asai S. Asai N.A. Asbah J. Assahsah K. Assamagan R. Astalos S. Atashi R.J. Atkin m. Atkinson H. Atmani P.A. Atmasiddha K. Augsten S. Auricchio A.D. Auriol V.A. Austrup G. Avolio K. Axiotis G. Azuelos D. babal H. bachacou K. bachas A. bachiu F. backman A. badea P. bagnaia m. bahmani A.J. bailey V.R. bailey J.T. baines L. baines O.K. baker E. bakos D. bakshi Gupta V. balakrishnan R. balasubramanian E.m. baldin P. balek E. ballabene F. balli L.m. baltes W.K. balunas J. balz E. banas m. bandieramonte A. bandyopadhyay S. bansal L. barak m. barakat E.L. barberio D. barberis m. barbero m.Z. barel K.N. barends T. barillari m-S. barisits T. barklow P. baron D.A. baron mo The ATLAS Collaboration

The identification of jets originating from quarks and gluons,often referred to as quark/gluon tagging,plays an important role in various analyses performed at the Large Hadron Collider,as Standard model measurements and searches for new particles decaying to quarks often rely on suppressing a large gluon-induced *** paper describes the measurement of the efficiencies of quark/gluon taggers developed within the ATLAS Collaboration,using √s=13 TeV proton–proton collision data with an integrated luminosity of 140 fb^(-1) collected by the ATLAS *** taggers with high performances in rejecting jets from gluon over jets from quarks are studied:one tagger is based on requirements on the number of inner-detector tracks associated with the jet,and the other combines several jet substructure observables using a boosted decision tree.A method is established to determine the quark/gluon fraction in data,by using quark/gluon-enriched subsamples defined by the jet *** in tagging efficiency between data and simulation are provided for jets with transverse momentum between 500 GeV and 2 TeV and for multiple tagger working points.

关键词： ATLAS JET QUARK GLUON TAGGING

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Improving wood volume predictions in dry tropical forest in the semi-arid brazil

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Journal of Arid Land 2020年第6期12卷 1046-1055页

作者： Robson b de LImA Patrícia A b bARRETO-GARCIA Alessandro de PAULA Jhuly E S pereira Flávia F de CARVALHO Silvio H m GOmES Department of Forest Engineering State University of AmapáMacapá68900070Brazil Department of Forest Science State University of Southwest BahiaVitória da Conquista 45083900Brazil Universidade Federal de Lavras Lavras 37200900Brazil Universidade de São Paulo Piracicaba 13418900Brazil

The volumetric variability of dry tropical forests in brazil and the scarcity of studies on the subject show the need for the development of techniques that make it possible to obtain adequate and accurate wood volume *** this study,we analyzed a database of thinning trees from a forest management plan in the Contendas de SincoráNational Forest,southwestern bahia State,*** data set included a total of 300 trees with a trunk diameter ranging from 5 to 52 ***,validation and statistical selection of four volumetric models were *** to the difference in height values for the same diameter and the low correlation between both variables,we do not suggest models which only use the diameter at breast height(DbH)variable as a predictor because they accommodate the largest estimation *** comparing the best single entry model(Hohenald-Krenn)with the Spurr model(best fit model),it is noted that the exclusion of height as a predictor causes the values of 136.44 and 0.93 for Akaike information criterion(AIC)and adjusted determination coefficient(R2 adj),which are poorer than the second best model(Schumacher-Hall).Regarding the minimum sample size,errors in estimation(root mean square error(RmSE)and bias)of the best model decrease as the sample size increases,especially when a larger number of trees with DbH≥15.0 cm are randomly *** sampling by diameter class produces smaller volume prediction errors than random sampling,especially when considering all *** summary,the Spurr and Schumacher-Hall models perform *** models suggest that the total variance explained in the estimates is not less than 95%,producing reliable forecasts of the total volume with *** estimates indicate that the bias around the average is not greater than 7%.Our results support the decision to use regression methods to build models and estimate their parameters,seeking stratification strategies in diameter classes for the sample trees.

关键词： volume modeling minimal sample size Caatinga Spurr model forest management

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Imidazolium-based ionic liquids as additives to preserve the Enhanced Green Fluorescent Protein fluorescent activity

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Green Chemical Engineering 2021年第4期2卷 412-422页

作者： Nathalia V.Veríssimo Carolina F.Saponi Timothy m.Ryan Tamar L.Greaves Jorge F.b.pereira Department of Engineering of bioprocesses and biotechnology School of Pharmaceutical SciencesSao Paulo State University(UNESP)Rodovia Araraquara-Jaú/Km 0114800-903AraraquaraSPBrazil School of Science College of ScienceEngineering and HealthRMIT University124 La Trobe StreetMelbourneVIC 3000Australia School of Pharmaceutical Sciences Sao Paulo University(USP)Av.Prof.Lineu Prestesno.580B1605508-000Cidade de UniversitariaSao PauloSPBrazil Australian Synchrotron 800 Blackburn RoadClaytonVIC 3168Australia Univ Coimbra CIEPQPFDepartment of Chemical EngineeringRua Sílvio LimaPolo II–Pinhal de Marrocos3030-790 CoimbraPortugal

Fluorescent Proteins(FP)can be applied as biomarkers and biosensors in the industrial and medical fields,but their large-scale use,especially for new industrial applications,is limited due to their low ***,the discovery of additives capable of preserving the activity of FP at room temperature and under stress conditions can help to expand and facilitate their commercial *** this goal,we evaluated the application of 1-alkyl-3-methylimidazolium chloride-based([C_(n)mim]Cl)ionic liquids(ILs)as additives to preserve the activity of Enhanced Green Fluorescent Protein(EGFP)at different storage times and under unfavorable ***[C_(n)mim]Cl solutions(at 0.100 mol L^(-1))were able to preserve EGFP fluorescence for longer than the phosphatesaline buffer(PbS)and NaCl solutions,increasing its fluorescence manifestation from 1 to 3 months.[C_(n)mim]Cl with shorter to medium cationic alkyl chains were the most effective in preserving EGFP fluorescence.[C_(n)mim]Cl also protected EGFP activity in the presence of the surfactant SDS,the acid guanidine hydrochloride,and H_(2)O_(2).Therefore,[C_(n)mim]Cl can be added to aqueous solutions to preserve EGFP fluorescence activity at room temperature for longer storage times and to reduce the negative impact of denaturing agents on ***,there is a massive potential for the application of ILs as additives to preserve FP in the long-term without refrigeration and under unfavorable conditions,and this is fundamental to enable expansion of FP in industrial and commercial applications.

关键词： Green Fluorescent Protein Protein stability Fluorescence Ionic liquids Preservatives

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