Surface plasmon resonance as a tool in the functional analisys of an immunodominant site in foot-and-mouth disease virus

A fast and direct surface plasmon resonance (SPR) method for the kinetic analysis of the interactions between peptide antigens and immobilised monoclonal antibodies (mAb) has been established. Protocols have been developed to overcome the problems posed by the small size of the analytes (< 1600 Da). The interactions were well described by a simple 1:1 bimolecular interaction and the rate constants were self-consistent and reproducible. The key features for the accuracy of the kinetic constants measured were high buffer flow rates, medium antibody surface densities and high peptide concentrations. The method was applied to an extensive analysis of over 40 peptide analogues towards two distinct anti-FMDV antibodies, providing data in total agreement with previous competition ELISA experiments. Eleven linear 15-residue synthetic peptides, reproducing all possible combinations of the four replacements found in foot-and-mouth disease virus (FMDV) field isolate C-S30, were evaluated. The direct kinetic SPR analysis of the interactions between these peptides and three anti-site A mAbs suggested additivity in all combinations of the four relevant mutations, which was confirmed by parallel ELISA analysis. The four-point mutant peptide (A15S30) reproducing site A from the C-S30 strain was the least antigenic of the set, in disagreement with previously reported studies with the virus isolate. Increasing peptide size from 15 to 21 residues did not significantly improve antigenicity. Overnight incubation of A15S30 with mAb 4C4 in solution showed a marked increase in peptide antigenicity not observed for other peptide analogues, suggesting that conformational rearrangement could lead to a stable peptide-antibody complex. In fact, peptide cyclization clearly improved antigenicity, confirming an antigenic reversion in a multiply substituted peptide. Solution NMR studies of both linear and cyclic versions of the antigenic loop of FMDV C-S30 showed that structural features previously correlated with antigenicity were more pronounced in the cyclic peptide. Twenty-six synthetic peptides, corresponding to all possible combinations of five single-point antigenicity-enhancing replacements in the GH loop of FMDV C-S8c1, were also studied. SPR kinetic screening of these peptides was not possible due to problems mainly related to the high mAb affinities displayed by these synthetic antigens. Solution affinity SPR analysis was employed and affinities displayed were generally comparable to or even higher than those corresponding to the C-S8c1 reference peptide A15. The NMR characterisation of one of these multiple mutants in solution showed that it had a conformational behaviour quite similar to that of the native sequence A15 and the X-ray diffraction crystallographic analysis of the peptide ? mAb 4C4 complex showed paratope ? epitope interactions identical to all FMDV peptide ? mAb complexes studied so far. Key residues for these interactions are those directly involved in epitope ? paratope contacts (141Arg, 143Asp, 146His) as well as residues able to stabilise a particular peptide global folding. A quasi-cyclic conformation is held up by a hydrophobic cavity defined by residues 138, 144 and 147 and by other key intrapeptide hydrogen bonds, delineating an open turn at positions 141, 142 and 143 (corresponding to the Arg-Gly-Asp motif).

Site de escola, uma janela aberta para o mundo

Revista Lusófona de Educação

Application of GIS-based multi-criteria analysis for site selection of aquifer recharge with reclaimed water

Reclaimed water from small wastewater treatment facilities in the rural areas of the Beira Interior region (Portugal) may constitute an alternative water source for aquifer recharge. A 21-month monitoring period in a constructed wetland treatment system has shown that 21,500 m(3) year(-1) of treated wastewater (reclaimed water) could be used for aquifer recharge. A GIS-based multi-criteria analysis was performed, combining ten thematic maps and economic, environmental and technical criteria, in order to produce a suitability map for the location of sites for reclaimed water infiltration. The areas chosen for aquifer recharge with infiltration basins are mainly composed of anthrosol with more than 1 m deep and fine sand texture, which allows an average infiltration velocity of up to 1 m d(-1). These characteristics will provide a final polishing treatment of the reclaimed water after infiltration (soil aquifer treatment (SAT)), suitable for the removal of the residual load (trace organics, nutrients, heavy metals and pathogens). The risk of groundwater contamination is low since the water table in the anthrosol areas ranges from 10 m to 50 m. Oil the other hand, these depths allow a guaranteed unsaturated area suitable for SAT. An area of 13,944 ha was selected for study, but only 1607 ha are suitable for reclaimed water infiltration. Approximately 1280 m(2) were considered enough to set up 4 infiltration basins to work in flooding and drying cycles.

A perspective on LCA application in site remediation services: critical review of challenges

The remediation of contaminated sites supports the goal of sustainable development but may also have environmental impacts at a local, regional and global scale. Life cycle assessment (LCA) has increasingly been used in order to support site remediation decision-making. This review article discusses existing LCA methods and proposed models focusing on critical decisions and assumptions of the LCA application to site remediation activities. It is concluded that LCA has limitations as an adequate holistic decisionmaking tool since spatial and temporal differentiation of non-global impacts assessment is a major hurdle in site remediation LCA. Moreover, a consequential LCA perspective should be adopted when the different remediation services to be compared generate different site’s physical states, displacing alternative post-remediation scenarios. The environmental effects of the post-remediation stage of the site is generally disregarded in the past site remediation LCA studies and such exclusion may produce misleading conclusions and misdirected decision-making. In addition, clear guidance accepted by all stakeholders on remediation capital equipment exclusion and on dealing with multifunctional processes should be developed for site remediation LCA applications.

Um site, um conteúdo e incontáveis dispositivos: as estratégias de design utilizadas para ambientes multiplataformas

Dissertação apresentada à Escola Superior de Comunicação Social como parte dos requisitos para obtenção de grau de mestre em Audiovisual e Multimédia.

Soils of the Jurassic formations in the central Algarve; their occurrence, distribution, genesis and site qualities

In the central Algarve different soils have developed dependent on petrography of the parent material, slope position and recent erosion. The general patterns of occurence and distribution of different soils are described. The age of an eutric Nitosol is estimated and the relation between the soil and the parent material is investigated. Some different soils are described as examples with their chemical and physical properties. The water budget of soils is described in general with considerations concerning ground water recharge and run-off as well as in dependence of climate and of different site conditions.

Assessment of climate change statistical downscaling methods: Application and comparison of two statistical methods to a single site in Lisbon

Dissertação apresentada na Faculdade de Ciências e Tecnologia da Universidade Nova de Lisboa para a obtenção do grau de Mestre em Engenharia do Ambiente

A high level e-maintenance architecture to support on-site teams

Emergent architectures and paradigms targeting reconfigurable manufacturing systems increasingly rely on intelligent modules to maximize the robustness and responsiveness of modern installations. Although intelligent behaviour significantly minimizes the occurrence of faults and breakdowns it does not exclude them nor can prevent equipment’s normal wear. Adequate maintenance is fundamental to extend equipments’ life cycle. It is of major importance the ability of each intelligent device to take an active role in maintenance support. Further this paradigm shift towards “embedded intelligence”, supported by cross platform technologies, induces relevant organizational and functional changes on local maintenance teams. On the one hand, the possibility of outsourcing maintenance activities, with the warranty of a timely response, through the use of pervasive networking technologies and, on the other hand, the optimization of local maintenance staff are some examples of how IT is changing the scenario in maintenance. The concept of e-maintenance is, in this context, emerging as a new discipline with defined socio-economic challenges. This paper proposes a high level maintenance architecture supporting maintenance teams’ management and offering contextualized operational support. All the functionalities hosted by the architecture are offered to the remaining system as network services. Any intelligent module, implementing the services’ interface, can report diagnostic, prognostic and maintenance recommendations that enable the core of the platform to decide on the best course of action.

Development of a meso-microscale coupling procedure for site assessment in complex terrain

A procedure for coupling mesoscale and CFD codes is presented, enabling the inclusion of realistic stratification flow regimes and boundary conditions in CFD simulations of relevance to site and resource assessment studies in complex terrain. Two distinct techniques are derived: (i) in the first one, boundary conditions are extracted from mesoscale results to produce time-varying CFD solutions; (ii) in the second case, a statistical treatment of mesoscale data leads to steady-state flow boundary conditions believed to be more representative than the idealised profiles which are current industry practice. Results are compared with measured data and traditional CFD approaches.

Finding Aedes aegypti in a natural breeding site in an urban zone, Sao Paulo, Southeastern Brazil

ABSTRACT This is the description of how nine Aedes aegypti larvae were found in a natural breeding site in the Pinheiros neighborhood, city of Sao Paulo, SP, Southeastern Brazil. The record was conducted in December 2014, during an entomological surveillance program of dengue virus vectors, with an active search of potential breeding sites, either artificial or natural. Finding Ae. aegypti larvae in a tree hole shows this species’ ability to use both artificial and natural environments as breeding sites and habitats, which points towards the importance of maintaining continuous surveillance on this mosquito in all kinds of water-holding containers.

Industry energy optimization: A case study in a biodiesel production site

This paper presents a case study of heat exchanger network (HEN) retrofit with the objective to reduce the utilities consumption in a biodiesel production process. Pinch analysis studies allow determining the minimum duty utilities as well the maximum of heat recovery. The existence of heat exchangers for heat recovery already running in the process causes a serious restriction for the implementation of grassroot HEN design based on pinch studies. Maintaining the existing HEN, a set of alternatives with additional heat exchangers was created and analysed using some industrial advice and selection criteria. The final proposed solution allows to increase the actual 18 % of recovery heat of the all heating needs of the process to 23 %, with an estimated annual saving in hot utility of 35 k(sic)/y.

Benchmarking de identificação de ano: criação de um site para benchmarking de técnicas de identificação de dano

Dissertação para obtenção do grau de Mestre em Engenharia Mecânica na Área de Manutenção e Produção

Experimental paracoccidioidomycosis in hamster: transmission electron microscopy of inoculation site lesion

Interaction between Paracoccidioides brasiliensis (Pb) and inflammatory cells in hamster testis was studied sequentially by transmission electron microscopy. In early lesions (six hours after inoculation), polymorphonuclear neutrophils (PMNs) were the major and mononuclear cells and eosinophils were the minor constituents of the inflammatory cells. PMNs were later replaced by mononuclear cells. Viable Pb cells were phagocytosed or surrounded by inflammatory cells. Preserved Pb cells usually had broad host-parasite interphases, whereas dying ones had narrow interphases. The outer layer of the fungus wall was sometimes broken by PMN in some focal points, broken pieces being peeled off and phagocytosed. Small Pb cells were uninuclear, and were often related to broad interphase. Large Pb cells were multinucleated with irregularly shaped wall, and sometimes had lomasome and/or myelin like structures. Different interaction patterns of Pb with inflammatory cells may be due to functionally different host cell flow to the inoculation site or due to the age of Pb cells or both.

The hamster cheek pouch: an immunologically privileged site suitable to the study of granulomatous infections

The hamster check pouch is an invagination of oral mucosa, characterized histologically as skin-like. In this paper we describe anatomical, histological and embriological features of the pouch and coment on the pouch as an immunologically privileged site since it lacks lymphatic drainage and has few Langerhans cells. We present the review from literature and our observations after inoculation in the pouch of mycobacteriae (BCG, Mycobacterium tuberculosis and Mycobacterium leprae) and a fungus (Paracoccidioides brasiliensis). Lesions in the pouch were granulomatous but smaller and long lasting; even granulomatous, the reaction was inefficient to control the proliferation of agents compared with inoculation in other sites, except for BCG. Appearance of immunity was also delayed or absent and, when it was detected, a sharp decrease in number of agents in pouch lesions was observed. These observations make the pouch an interesting site for the study of the role of immune system in infeccious diseases and in granuloma formation.