RESUMO
During the secondary production of aluminum, upon melting the scrap in a furnace, there is the possibility of developing an aluminothermic reaction, which produces oxides in the molten metal bath. Aluminum oxides must be identified and removed from the bath, as they modify the chemical composition and reduce the purity of the product. Furthermore, accurate measurement of molten aluminum level in a casting furnace is crucial to obtain an optimal liquid metal flow rate which influences the final product quality and process efficiency. This paper proposes methods for the identification of aluminothermic reactions and molten aluminum levels in aluminum furnaces. An RGB Camera was used to acquire video from the furnace interior, and computer vision algorithms were developed to identify the aluminothermic reaction and melt level. The algorithms were developed to process the image frames of video acquired from the furnace. Results showed that the proposed system allowed the online identification of the aluminothermic reaction and the molten aluminum level present inside the furnace at a computation time of 0.7 s and 0.4 s per frame, respectively. The advantages and limitations of the different algorithms are presented and discussed.
Assuntos
Óxido de Alumínio , Alumínio , Alumínio/química , Óxidos/químicaRESUMO
Chicken avidin and bacterial streptavidin are widely employed in vitro for their capacity to bind biotin, but their pharmacokinetics and immunological properties are not always optimal, thereby limiting their use in medical treatments. Here we investigate the biochemical and biological properties of a new modified avidin, obtained by ligand-assisted sodium periodate oxidation of avidin. This method allows protection of biotin-binding sites of avidin from inactivation caused by the oxidation step and delay of avidin clearance from injected tissue by generation of aldehyde groups from avidin carbohydrate moieties. Oxidized avidin shows spectroscopic properties similar to that of native avidin, indicating that tryptophan residues are spared from oxidation damage. In strict agreement with these results, circular dichroism and isothermal titration calorimetry analyses confirm that the ligand-assisted oxidation preserves the avidin protein structure and its biotin binding capacity. In vitro cell binding and in vivo tissue residence experiments demonstrate that aldehyde groups provide oxidized avidin the property to bind cellular and interstitial protein amino groups through Schiff's base formation, resulting in a tissue half-life of 2 weeks, compared with 2 h of native avidin. In addition, the efficient uptake of the intravenously injected (111)In-BiotinDOTA (ST2210) in the site previously treated with modified avidin underlines that tissue-bound oxidized avidin retains its biotin binding capacity in vivo. The results presented here indicate that oxidized avidin could be employed to create a stable artificial receptor in diseased tissues for the targeting of biotinylated therapeutics.
Assuntos
Avidina/química , Bioquímica/métodos , Aldeídos/química , Animais , Biotinilação/métodos , Galinhas , Feminino , Citometria de Fluxo , Ligantes , Camundongos , Camundongos Endogâmicos BALB C , Oxigênio/química , Ácido Periódico/química , Ligação Proteica , Espectrofotometria Ultravioleta/métodosRESUMO
Using a combinatorial chemistry approach, we identified a tetrameric tripeptide, denoted Protein A Mimetic (PAM) or TG19318, able to bind to immunoglobulins of different classes and species. The inverso variant, with the tripeptide in the all-D configuration (D-PAM or TG19320), is described as retaining binding properties to Ig. This peptide has now been assayed as a binder for E class immunoglobulins, in linear and competitive ELISA experiments, dot-blot and surface plasmon resonance (SPR) analyses. We show that D-PAM binds IgE with high specificity and selectivity, the interaction being sufficient to inhibit anaphylactic release of beta-hexosaminidase from RBL 2H3 cells, with an IC50 value of 10 microg/mL. Intradermal administration of D-PAM suppresses PCA in the rat, with an IC50 of 1.25 microg/kg dose of peptide, while its intraperitoneal injection inhibits mouse PCA with an IC50 of about 7 mg/kg and an efficacy comparable to that of ketotifen. Similarly, D-PAM inhibits ACA in the mouse, with 50% suppression at 10 mg/kg. The results presented here show that the peptide is active on the studied models, with effective doses below toxicity level, hence the molecule is a promising candidate for development of a new class of anti-allergic drugs.
Assuntos
Antialérgicos/farmacologia , Imunoglobulina E/metabolismo , Peptídeos/química , Peptídeos/farmacologia , 2,4-Dinitrofenol , Animais , Biotinilação , Hexosaminidases/metabolismo , Humanos , Camundongos , Anafilaxia Cutânea Passiva/imunologia , Ligação Proteica/efeitos dos fármacos , Conformação Proteica/efeitos dos fármacos , Ratos , Ressonância de Plasmônio de SuperfícieRESUMO
In recent years, the study of decision making has provided a paradigmatic case of "crossbreeding" of different disciplines. The integration of economics, psychology and neurosciences within neuroeconomics calls for more accurate and comprehensive models of human rationality, which may be obtained by combining diverse theoretical approaches and experimental techniques. In this respect, neuroeconomics contributes to a naturalistic, brain-based, explanation of human agency. However, although contemporary naturalism insists on the unitary aspect of reality, we stress that supporting unitary study of nature is not the same as supporting a single, fundamental discipline to which all higher-order analyses could (or should) be reduced. We argue for integration, rather than reduction, as the best approach to a naturalistic explanation of human decision making, and we claim that supporting epistemological pluralism does not mean being committed to any specific ontological position. However, we suggest that an "emergentist" ontology is the best candidate to integrate the epistemological analysis here endorsed.
Assuntos
Encéfalo/fisiologia , Tomada de Decisões/fisiologia , Conhecimento , Neurobiologia/métodos , Humanos , Modelos Neurológicos , Neurofisiologia , Filosofia Médica , Teoria PsicológicaRESUMO
Esterified hyaluronic acid (HYAFF) is routinely used for clinical tissue-engineering applications such as skin and cartilage. The material is degraded by neotissue formation and degradation products are highly biocompatible. In the present article we investigate the possibility to culture vascular smooth muscle cells on this biodegradable material for the generation of tubular constructs to be used for vascular tissue engineering. We have evaluated cell attachment and growth, and the possibility to obtain a three-dimensional tubular shape culture from flat HYAFF sheets. We also evaluated the mechanical properties of the cell constructs, using a specific testing protocol, and compared them with the properties of segments of porcine coronary artery. Morphology and viability tests demonstrated that vascular cells, either from porcine or human origin, adhere and grow on nonwoven meshes of HYAFF, and that precoating of the material with fibronectin or collagen had a modest effect on cell growth and extracellular matrix production. Cell growth reached a maximum 7 days after seeding. Simple wrapping of flat sheets of nonwoven meshes containing vascular cells around a cylindrical mandrel, and culture under static conditions for 14 days, yielded tubular constructs suitable for mechanical tests. Despite cell colonization, constructs showed lower mechanical resistance as compared with porcine coronary arteries. The material used and the technique developed result in highly cellularized tubular constructs. Whether the mechanical properties may be improved by dynamic culture conditions is worthy of investigation.
Assuntos
Prótese Vascular , Técnicas de Cultura de Células/métodos , Células Endoteliais/citologia , Células Endoteliais/fisiologia , Ácido Hialurônico , Engenharia Tecidual/métodos , Animais , Materiais Biocompatíveis , Adesão Celular/fisiologia , Proliferação de Células , Tamanho Celular , Sobrevivência Celular/fisiologia , Células Cultivadas , Vasos Coronários/fisiologia , Elasticidade , Teste de Materiais , Membranas Artificiais , Suínos , Resistência à Tração/fisiologiaRESUMO
Intravascular stents are small tube-like structures expanded into stenotic arteries to restore blood flow perfusion to the downstream tissues. The stent is mounted on a balloon catheter and delivered to the site of blockage. When the balloon is inflated, the stent expands and is pressed against the inner wall of the coronary artery. After the balloon is deflated and removed, the stent remains in place, keeping the artery open. Hence, the stent expansion defines the effectiveness of the surgical procedure: it depends on the stent geometry, it includes large displacements and deformations and material non-linearity. In this paper, the finite element method is applied (i) to understand the effects of different geometrical parameters (thickness, metal-to-artery surface ratio, longitudinal and radial cut lengths) of a typical diamond-shaped coronary stent on the device mechanical performance, (ii) to compare the response of different actual stent models when loaded by internal pressure and (iii) to collect suggestions for optimizing the device shape and performance. The stent expansion and partial recoil under balloon inflation and deflation were simulated. Results showed the influence of the geometry on the stent behavior: a stent with a low metal-to-artery surface ratio has a higher radial and longitudinal recoil, but a lower dogboning. The thickness influences the stent performance in terms of foreshortening, longitudinal recoil and dogboning. In conclusion, a finite element analysis similar to the one herewith proposed could help in designing new stents or analyzing actual stents to ensure ideal expansion and structural integrity, substituting in vitro experiments often difficult and unpractical.
Assuntos
Simulação por Computador , Estenose Coronária/cirurgia , Análise de Falha de Equipamento/métodos , Modelos Cardiovasculares , Stents , Elasticidade , Análise de Elementos Finitos , Humanos , Pressão , Desenho de Prótese , Sensibilidade e Especificidade , Estresse MecânicoRESUMO
The evaluation of contact areas and pressures in total knee prosthesis is a key issue to prevent early failure. The first part of this study is based on the hypothesis that the patterns of contact stresses on the tibial insert of a knee prosthesis at different stages of the gait cycle could be an indicator of the wear performances of a knee prosthesis. Contact stresses were calculated for a mobile bearing knee prosthesis by means of finite element method (FEM). Contact areas and stresses were also measured through in vitro tests using Fuji Prescale film in order to support the FEM findings. The second part of this study addresses the long-term structural integrity of metal tibial components in terms of fatigue life by means of experimental tests and FEM simulations. Fatigue experimental evaluations were performed on Cr-Co alloy tibial tray, based on ISO standards. FEM models were used to calculate the stress patterns. The failure risk was estimated with a standard fatigue criterion on the basis of the results obtained from the FEM calculations. Experimental and computational results showed a positive matching.
Assuntos
Análise de Falha de Equipamento/métodos , Marcha , Articulação do Joelho/fisiopatologia , Articulação do Joelho/cirurgia , Prótese do Joelho , Modelos Biológicos , Falha de Prótese , Suporte de Carga , Simulação por Computador , Desenho Assistido por Computador , Análise de Elementos Finitos , Humanos , Pressão , Desenho de Prótese/métodos , Estresse MecânicoRESUMO
The most promising approach in Alzheimer disease immunotherapy is represented by amyloid beta derivatives with low intrinsic neurotoxicity and minimal overall T cell responses. To avoid toxicity and autoimmune response, we have designed a new class of Abeta derivatives through segmentation of the original Abeta[1-42] peptide and application of the glycine substitution modification technology. Abeta[1-16], Abeta[13-28] and Abeta[25-42] fragments were selected in order to retain the major immunogenic sites of the Abeta[1-42] peptide. All peptides showed comparable immunogenicity, and raised antibodies were all able to cross-recognize both Abeta[1-42] and Abeta[1-40] synthetic amyloid forms. Polyclonal antibodies produced against the simplified variants were able to recognize the parent peptide, but not the opposite simplified forms, in strict agreement with the model of independent surfaces of recognition. All Abeta simplified derivatives showed reduced fibrillogenic properties, thus underlining that the introduction of glycine residues in alternating positions allows to obtain modified peptides maintaining the main immunogenic properties of the parent peptides, but with reduced ability to adopt a beta-sheet conformation and therefore a much lower risk of toxicity in humans. In addition, in vitro studies on peripheral blood mononuclear cells (PBMCs) from healthy donors showed that only the Abeta[13-28]+G peptide failed to induce IFN-gamma production, thus suggesting that this molecule could represent a good candidate for potentially safer vaccine therapy to reduce amyloid burden in Alzheimer's disease instead of using toxic Abeta[1-42].
Assuntos
Peptídeos beta-Amiloides/imunologia , Oligopeptídeos/imunologia , Fragmentos de Peptídeos/imunologia , Sequência de Aminoácidos , Amiloide/química , Amiloide/imunologia , Peptídeos beta-Amiloides/química , Animais , Anticorpos/imunologia , Especificidade de Anticorpos/imunologia , Linhagem Celular , Células Cultivadas , Reações Cruzadas/imunologia , Ensaio de Imunoadsorção Enzimática , Citometria de Fluxo , Humanos , Imunização , Imunoglobulina G/imunologia , Interferon gama/metabolismo , Leucócitos Mononucleares/citologia , Leucócitos Mononucleares/efeitos dos fármacos , Leucócitos Mononucleares/imunologia , Ativação Linfocitária/efeitos dos fármacos , Ativação Linfocitária/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Dados de Sequência Molecular , Oligopeptídeos/síntese química , Oligopeptídeos/farmacologia , Fragmentos de Peptídeos/farmacologia , Linfócitos T/citologia , Linfócitos T/imunologia , Linfócitos T/metabolismoRESUMO
PTX3 is a secreted multimeric glycoprotein which plays a key role in innate immunity by activating the classical complement pathway through specific recognition of the C1q subunit. A method is described for the high level expression of the recombinant human PTX3 in Chinese hamster ovary cells (CHO), adapted to a suspension growth in spinner flasks containing a serum-free chemically defined medium and producing about 50 mg of PTX3/L of culture. A purification procedure to produce a homogeneous protein preparation from the supernatant, by means of anion exchange, hydroxyapatite and size exclusion chromatography, is also reported. This three-step protocol allows us to obtain PTX3 with a recovery yield close to 70%, a purity degree exceeding 95%, and a final host cell protein (HCP) content lower than 150 ppm. The recombinant purified PTX3 retains its biological activity, as demonstrated by C1q binding ELISA assay, and displays a complex quaternary structure characterized by a high secondary structure content quite different from human short pentraxin C-reactive protein (CRP) and serum amyloid P component (SAP), as determined by circular dichroism, fluorescence analysis, and native and SDS-PAGE experiments.