Role of inflammatory mediators in priming, activation, and deformability of bovine neutrophils

Objective-To determine the capacity of inflammatory mediators tumor necrosis factor-alpha (TNF-alpha), interleukin-8 (IL-8), platelet-activating factor (PAF), lipopolysaccharide (LPS), and leukotoxin to prime, activate, or alter deformability of adult bovine neutrophils.Sample Population-Blood collected from 5 healthy adult Holstein cows.Procedure-Isolated neutrophils or whole brood was incubated with TNF-alpha, IL-8, PAF, LPS, or leukotoxin, and neutrophil chemiluminescence, degranulation, deformability, shape change, CD11b expression, and size distribution was measured.Results-incubation with TNF-alpha, IL-8; PAF, and IFS primed neutrophils for oxygen radical release but caused minimal oxygen radical release by themselves. None of the inflammatory mediators induced degranulation. Incubation with TNF-alpha and PAF resulted in a decrease in neutrophil deformability and induced shape change in neutrophils. incubation with PAF consistently resulted in an increase in neutrophil size as measured by use of flow cytometry. Only IL-8 caused an increase in expression of CD11b by neutrophils.Conclusions and Clinical Relevance-Inflammatory mediators tested had minimal effects on neutrophil oxygen radical production or degranulation but did prime neutrophils for oxygen radical production. Incubation with PAF and TNF-alpha caused a decrease in neutrophil deformability and altered neutrophil shape and size. Results of our study indicate that PAF- and TNF-alpha-induced changes in neutrophil deformability and size may cause integrin- and setectin-independent trapping of neutrophils in the lungs of cattle with pneumonic pasteurellosis.

Active and exo-site inhibition of human factor Xa: Structure of des-Gla factor Xa inhibited by NAP5, a potent nematode anticoagulant protein from Ancylostoma caninum

Hookworms are hematophagous nematodes capable of growth, development and subsistence in living host systems such as humans and other mammals. Approximately one billion, or one in six, people worldwide are infected by hookworms causing gastrointestinal blood loss and iron deficiency anemia. The hematophagous hookworm Ancylostoma caninum produces a family of small, disulfide-linked protein anticoagulants (75-84 amino acid residues). One of these nematode anticoagulant proteins, NAP5, inhibits the amidolytic activity of factor Xa (fXa) with K-i = 43 pM, and is the most potent natural fXa inhibitor identified thus far. The crystal structure of NAP5 bound at the active site of gamma-carboxyglutamic acid domainless factor Xa (des-fXa) has been determined at 3.1 angstrom resolution, which indicates that Asp189 (fXa, S1 subsite) binds to Arg40 (NAP5, P1 site) in a mode similar to that of the BPTI/trypsin interaction. However, the hydroxyl group of Ser39 of NAP5 additionally forms a hydrogen bond (2.5 angstrom) with His57 NE2 of the catalytic triad, replacing the hydrogen bond of Ser195 OG to the latter in the native structure, resulting in an interaction that has not been observed before. Furthermore, the C-terminal extension of NAP5 surprisingly interacts with the fXa exosite of a symmetry-equivalent molecule forming a short intermolecular beta-strand as observed in the structure of the NAPc2/fXa complex. This indicates that NAP5 can bind to fXa at the active site, or the exosite, and to fX at the exosite. However, unlike NAPc2, NAP5 does not inhibit fVIIa of the fVIIa/TF complex. (c) 2007 Elsevier Ltd. All rights reserved.

Determination of site-site distance and site concentration within polymer beads: A combined swelling-electron paramagnetic resonance study

[GRAPHICS]This work proposes a combined swelling-electron paramagnetic resonance (EPR) approach aiming at determining some unusual polymer solvation parameters relevant for chemical processes occurring inside beads. Batches of benzhydrylamine-resin (BHAR), a copolymer of styrene-1% divinylbenzene containing phenylmethylamine groups were, labeled with the paramagnetic amino acid 2,2,6,6-tetramethylpiperidine-1-oxyl-4-amine-4-carboxylic acid (TOAC), and their swelling properties and EPR spectra were examined in DCM and DMF. By taking into account the BHARs labeling degrees, the corresponding swelling values, and some polymer structural characteristics, it was possible to calculate polymer swelling parameters, among them, the volume and the number of sites per bead, site-site distances and site concentration. The latter values ranged from 17 to 170 angstrom and from 0.4 to 550 mM, respectively. EPR spectroscopy was applied to validate the multistep calculation strategy of these swelling parameters. Spin-spin interaction was detected in the labeled resins at site-site distances less than approximately 60 A or probe concentrations higher than approximately 1 x 10(-2) M, in close agreement with the values obtained for the spin probe free in solution. Complementarily, the yield of coupling reactions in different resins indicated that the greater the inter-site distance or the lower the site concentration, the faster the reaction. The results suggested that the model and the experimental measurements developed for the determination of solvation parameters represent a relevant step forward for the deeper understanding and improvement of polymer-related processes.

Métodos lineares e não lineares de análise de séries temporais e sua aplicação no estudo da variabilidade da frequência cardíaca de jovens saudáveis

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Papel dos receptores Toll-Like na atividade dos neutrófilos humanos desafiados com a cepa de alta virulência do Paracoccidioides brasiliensis

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Clonagem e expressão de fragmentos de anticorpos (scFV) contra o vírus da leucemia felina (FeLV) por phage display

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Ontopolíticas e diagramas históricos do poder: maioria e minoria segundo Deleuze e a teoria das multidões segundo Peirce

This article aims at developing the so-called ontopolitics as G. Deleuze s innovative contribution to contemporary political philosophy. This objective will lead us to inspect the concept of power that Deleuze borrowed from Foucault and extended in order to assign to it an ontological adequacy. The concept of power opens access to another important element of the Deleuzean political philosophy, that is, the study of the historical diagrams of the power in the so-called discipline and control societies. With the combined dynamical diagram of both, we become aware of the portrait Deleuze draws for the democracy in contemporary societies. Digging into the Deleuzean ontopolitics, we will devote ourselves to the concepts of majority, minority and minor-becoming. It is in this point that the meeting between Deleuze s ontoplitics and Ch. Sanders Peirce s mathematical ontology becomes sound. It happens that Deleuze s ontopolitical concepts, besides their bond to an ontology of the power, receive also a mathematical treatment related to certain arithmetical (denumerable and nondenumerable) and geometrical notions (lines). The majorities and minorities are denumerable sets which are crossed by nondenumerable becomings. This step done, we will reach the stand point of the present paper, where we carry out initial approach with regard to an image for the concepts of majority and minority on the basis of Peirce s theory of collections and multitudes, mostly envisaging the mathematical ontology included in it. Accordingly, the main operation to be accomplished is that the Deleuzean distinction between the denumerable majorities/minorities and the nondenumerable mino-becoming may be mapped out in terms of discrete collections called enumerable, denumerable and abnumerable or postnumerable, in compliance with Peirce s terminology.

Dilemas da produção de conhecimento no campo acadêmico: entre especialização e difusão

The paper explores some issues related to the production of knowledge in the academic fi eld, relating it to the problem of knowledge dissemination beyond this restricted universe. Taking on theoretical sources from Bourdieu’s fi eld concept and his studies concerning the university fi eld in the work Homo academicus, seeks to examine, in an exploratory approach, the operation modes from academic and diffusion fi elds, opposing the actions from so-called experts and intellectual media, or intermediaries.

Morphometric evaluation by ultrasonographic exam of the portal veinm vaudal vein and abdominal aorta in healthy dogs of diferent body weights

The diameters and areas of portal vein, caudal vena cava and abdominal aorta are useful measurements in dogs. These values can be easily measured by ultrasonographic exam, and variations of normality can be an important indicator of hepatic or extra-hepatic alterations. This study aimed to measure the diameter and areas of portal vein, caudal vena cava and abdominal aorta inhealthy dogs, with normal corporal score, divided in groups according to the body weight, and assess whether the data are influenced by animal weight. Thirty dogs were examined and divided into three groups (Group A: ≤ 10 kg Group B: from 10.1 to 20.0 kg; Group C: ≥ 20.1 kg). To measure thediameters and areas of portal vein, caudal vena cava and abdominal aorta, the animal was kept in left lateral decubitus position and the transducer was placed on the right lateral abdominal wall, at approximately the 10th or 11th intercostal space, in the porta hepatis region. The diameters and areas of the portal vein, caudal vena cava and abdominal aorta were significantly lower for dogs in Group A with respect to other groups and the dogs from Groups B and C had similar results with each other. The diameters and areas of the portal vein, caudal vena cava and abdominal aorta may vary with the animal size, and reference values must be specific for small, medium and large dogs. Key words: abdominal vessels; area; diameter; measurement; ultrasonographic exam

Interleukin-10 production by tumor infiltrating macrophages plays a role in Human Papillomavirus 16 tumor growth

Abstract Background Human Papillomavirus, HPV, is the main etiological factor for cervical cancer. Different studies show that in women infected with HPV there is a positive correlation between lesion grade and number of infiltrating macrophages, as well as with IL-10 higher expression. Using a HPV16 associated tumor model in mice, TC-1, our laboratory has demonstrated that tumor infiltrating macrophages are M2-like, induce T cell regulatory phenotype and play an important role in tumor growth. M2 macrophages secrete several cytokines, among them IL-10, which has been shown to play a role in T cell suppression by tumor macrophages in other tumor models. In this work, we sought to establish if IL-10 is part of the mechanism by which HPV tumor associated macrophages induce T cell regulatory phenotype, inhibiting anti-tumor activity and facilitating tumor growth. Results TC-1 tumor cells do not express or respond to IL-10, but recruit leukocytes which, within the tumor environment, produce this cytokine. Using IL-10 deficient mice or blocking IL-10 signaling with neutralizing antibodies, we observed a significant reduction in tumor growth, an increase in tumor infiltration by HPV16 E7 specific CD8 lymphocytes, including a population positive for Granzyme B and Perforin expression, and a decrease in the percentage of HPV specific regulatory T cells in the lymph nodes. Conclusions Our data shows that in the HPV16 TC-1 tumor mouse model, IL-10 produced by tumor macrophages induce regulatory phenotype on T cells, an immune escape mechanism that facilitates tumor growth. Our results point to a possible mechanism behind the epidemiologic data that correlates higher IL-10 expression with risk of cervical cancer development in HPV infected women.

O edifício da FAUUSP e os materiais do brutalismo

O trabalho analisa a história da produção do edifício da Faculdade de Arquitetura e Urbanismo da Universidade de São Paulo, projetado por João Batista Vilanova Artigas (1915-1985) em 1961 e concluído em 1969. Mais especificamente, o artigo se volta aos materiais do edifício, oferecendo insumos para uma discussão sobre o papel do chamado “brutalismo paulista” na década de 1960. O edifício da FAUUSP, como se sabe, é um marco na arquitetura moderna brasileira. Seu caráter paradigmático consiste na síntese das posições programáticas de Artigas, tanto em relação ao ensino de arquitetura como em relação à poética moderna, que ultrapassa o limite autoral e constitui uma escola. São características dessa escola – por vezes chamada de “paulista”, “brutalista” ou “artiguista” – alguns princípios como a continuidade espacial, o elogio das formas estruturais, a verdade dos materiais e o desenvolvimento das forças produtivas através da superação tecnológica. Esses princípios respondiam a algumas das questões mais urgentes da arquitetura moderna brasileira naquele período, deslocando a nova monumentalidade para uma dimensão construtiva. A nova poética, claramente exposta no projeto da FAUUSP, coincide com o surgimento de uma estética que atribuiu um novo valor político e crítico à produção. Passam a ser frequentes, por exemplo, interpretações do concreto armado brasileiro como síntese do subdesenvolvimento, por suas marcas de feitura artesanal e seus recordes tecnológicos. O objetivo deste artigo é contribuir com esse debate caracterizando em detalhe a produção do edifício em seus materiais: o concreto armado, a esquadria, os materiais de acabamento e os componentes industriais. O exame de aspectos históricos da produção do edifício, recolhidos em documentos originais e depoimentos, permite verificarmos de que modo a poética arquitetônica participa das decisões da obra e qual o seu impacto na economia do edifício e em sua forma de produção. Aprendemos, por exemplo, que o concreto armado apresenta manifestações visuais distintas e independentes de seu modo de produção (que foram pelo menos três: moldado in loco, protendido, e com agregados leves, sem função estrutural); que a empena da fachada exigiu uma fundação própria para seu cimbramento; que a modulação dos caixilhos não correspondia à modulação da estrutura; que a resina epóxi foi usada de modo pioneiro e experimental; que as fôrmas foram produzidas por um hábil carpinteiro português, mas as relações de trabalho eram precárias; que a contratação do projeto básico previa o detalhamento durante a obra pelo Escritório Técnico do Fundo para a Construção da Cidade Universitária. Essa noção ampla de material, entendida como a matéria mais o trabalho social que a define historicamente, nos permite tratar, simultaneamente, de aspectos técnicos, econômicos e artísticos da obra e assim compreender com maior exatidão dos termos do debate acerca da produção na arquitetura “brutalista” e seu papel político.

Interconnection systems for highly integrated computation devices

The sustained demand for faster,more powerful chips has beenmet by the availability of chip manufacturing processes allowing for the integration of increasing numbers of computation units onto a single die. The resulting outcome, especially in the embedded domain, has often been called SYSTEM-ON-CHIP (SOC) or MULTI-PROCESSOR SYSTEM-ON-CHIP (MPSOC). MPSoC design brings to the foreground a large number of challenges, one of the most prominent of which is the design of the chip interconnection. With a number of on-chip blocks presently ranging in the tens, and quickly approaching the hundreds, the novel issue of how to best provide on-chip communication resources is clearly felt. NETWORKS-ON-CHIPS (NOCS) are the most comprehensive and scalable answer to this design concern. By bringing large-scale networking concepts to the on-chip domain, they guarantee a structured answer to present and future communication requirements. The point-to-point connection and packet switching paradigms they involve are also of great help in minimizing wiring overhead and physical routing issues. However, as with any technology of recent inception, NoC design is still an evolving discipline. Several main areas of interest require deep investigation for NoCs to become viable solutions: • The design of the NoC architecture needs to strike the best tradeoff among performance, features and the tight area and power constraints of the on-chip domain. • Simulation and verification infrastructure must be put in place to explore, validate and optimize the NoC performance. • NoCs offer a huge design space, thanks to their extreme customizability in terms of topology and architectural parameters. Design tools are needed to prune this space and pick the best solutions. • Even more so given their global, distributed nature, it is essential to evaluate the physical implementation of NoCs to evaluate their suitability for next-generation designs and their area and power costs. This dissertation focuses on all of the above points, by describing a NoC architectural implementation called ×pipes; a NoC simulation environment within a cycle-accurate MPSoC emulator called MPARM; a NoC design flow consisting of a front-end tool for optimal NoC instantiation, called SunFloor, and a set of back-end facilities for the study of NoC physical implementations. This dissertation proves the viability of NoCs for current and upcoming designs, by outlining their advantages (alongwith a fewtradeoffs) and by providing a full NoC implementation framework. It also presents some examples of additional extensions of NoCs, allowing e.g. for increased fault tolerance, and outlines where NoCsmay find further application scenarios, such as in stacked chips.

Lesion detection and classification in breast cancer: evaluation of approaches based on morphological features, tracer kinetic modelling and semi-quantitative parameters in MR functional imaging (DCE-MRI)

The diagnosis, grading and classification of tumours has benefited considerably from the development of DCE-MRI which is now essential to the adequate clinical management of many tumour types due to its capability in detecting active angiogenesis. Several strategies have been proposed for DCE-MRI evaluation. Visual inspection of contrast agent concentration curves vs time is a very simple yet operator dependent procedure, therefore more objective approaches have been developed in order to facilitate comparison between studies. In so called model free approaches, descriptive or heuristic information extracted from time series raw data have been used for tissue classification. The main issue concerning these schemes is that they have not a direct interpretation in terms of physiological properties of the tissues. On the other hand, model based investigations typically involve compartmental tracer kinetic modelling and pixel-by-pixel estimation of kinetic parameters via non-linear regression applied on region of interests opportunely selected by the physician. This approach has the advantage to provide parameters directly related to the pathophysiological properties of the tissue such as vessel permeability, local regional blood flow, extraction fraction, concentration gradient between plasma and extravascular-extracellular space. Anyway, nonlinear modelling is computational demanding and the accuracy of the estimates can be affected by the signal-to-noise ratio and by the initial solutions. The principal aim of this thesis is investigate the use of semi-quantitative and quantitative parameters for segmentation and classification of breast lesion. The objectives can be subdivided as follow: describe the principal techniques to evaluate time intensity curve in DCE-MRI with focus on kinetic model proposed in literature; to evaluate the influence in parametrization choice for a classic bi-compartmental kinetic models; to evaluate the performance of a method for simultaneous tracer kinetic modelling and pixel classification; to evaluate performance of machine learning techniques training for segmentation and classification of breast lesion.

Pulse or hyaline ring granuloma. Review of the literature on etiopathogenesis of oral and extraoral lesions

Since the late 1950s, reports on an unusual giant-cell granulomatous lesion affecting the jaws, lungs, stomach and intestines have been published. Histopathologically, the lesions showed the presence of structureless hyaline rings with multinucleated giant cells. The aim of this review was to summarize the literature on the etiopathogenesis of the so-called oral and extraoral pulse or hyaline ring granuloma. Literature was searched using PubMed and Medline. In addition, hand search was performed. Search words were oral and extraoral hyaline ring granuloma, giant-cell hyaline angiopathy, pulse granuloma and chronic periostitis. Numerous terms for hyaline ring granuloma have been introduced over time (1971-2008). One hundred seventy-three cases of oral hyaline ring granuloma have been retrieved from the literature. In the mandible, 72.3% occurred . Two theories for etiopathogenesis have been proposed: (1) the origin of the hyaline rings is due to a foreign material (pulse and legumes) having penetrated the oral mucosa or gastrointestinal tract and lungs (exogenous theory) and (2) the rings are due to hyaline degenerative changes in walls of blood vessels (endogenous theory). Experimental production of oral and extraoral hyaline ring granulomas is consistent with the exogenous origin. Particles or remains of leguminous cells having been implanted or aspirated into human tissues whether located to the oral cavity or throughout the entire digestive tract and respiratory system are thought to be causative. Pulse or hyaline ring granulomas are rare but are well-defined oral and extraoral lesions due to implantation of the cellulose moiety of plant foods in contrast to the starch components.

Reduced incidence of insect-bite hypersensitivity in Icelandic horses is associated with a down-regulation of interleukin-4 by interleukin-10 and transforming growth factor-beta1

Insect bite hypersensitivity (IBH) is an allergic dermatitis of horses caused by IgE-mediated reactions to bites of insects of the genus Culicoides. IBH does not occur in Iceland due to the absence of Culicoides. However, Icelandic horses exported to mainland Europe as adults (1st generation) have a >/=50% incidence of developing IBH. In contrast, their progeny (2nd generation) has a <10% incidence of IBH. Here we show that peripheral blood mononuclear cells (PBMC) from Icelandic horses born in mainland Europe and belonging either to the IBH or healthy subgroup produce less interleukin (IL)-4 after polyclonal or allergen-specific stimulation when compared with counterparts from horses born in Iceland. We examined a role of IL-10 and transforming growth factor (TGF)-beta1 in down-regulation of IL-4 in healthy 2nd generation Icelandic horses. Supernatants of PBMC from 2nd generation healthy horses down-regulated the proportion of IL-4-producing cells and IL-4 production in stimulated cultures of PBMC from 1st generation IBH. This inhibition was mimicked by a combination of IL-10 and TGF-beta1 but not by the single cytokines. Cultures of stimulated PBMC of healthy 2nd generation horses produced a low level of IL-4, but IL-4 production was increased by anti-equine IL-10 and anti-human TGF-beta1. This shows for the first time that in horses, IL-10 and TGF-beta1 combined regulate IL-4 production in vitro. It is suggested that in this naturally occurring IgE-mediated allergy, IL-10 and TGF-beta1 have a role in the down-regulation of IL-4-induced allergen-specific Th2 cells, thereby reducing the incidence of IBH.