33 resultados para Constructs

em Repositório Institucional UNESP - Universidade Estadual Paulista "Julio de Mesquita Filho"


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The use of transposable elements (TEs) as genetic drive mechanisms was explored using Drosophila melanogaster as a model system. Alternative strategies, employing autonomous and nonautonomous P element constructs were compared for their efficiency in driving the ry(+) allele into populations homozygous for a ry(-) allele at the genomic rosy locus. Transformed flies were introduced at 1%, 5%, and 10% starting frequencies to establish a series of populations that were monitored over the course of 40 generations, using both phenotypic and molecular assays. The transposon-borne ry(+) marker allele spread rapidly in almost all populations when introduced at 5% and 10% seed frequencies, but 1% introductions frequently failed to become established. A similar initial rapid increase in frequency of the ry(+) transposon occurred in several control populations lacking a source of transposase. Constructs carrying ry(+) markers also increased to moderate frequencies in the absence of selection on the marker. The results of Southern and in situ hybridization studies indicated a strong inverse relationship between the degree of conservation of construct integrity and transposition frequency. These finding have relevance to possible future applications of transposons as genetic drive mechanisms.

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Background: Current classifications of Mental Disorders are centered on Westernized concepts and constructs. Cross-cultural sensitivity emphasizes culturally-appropriate translations of symptoms and questions, assuming that concepts and constructs are applicable.Methods: Groups and individual psychiatrists from various cultures from Asia, Latin America, North Africa and Eastern Europe prepared descriptions of main symptoms and complaints of treatment-seeking women in their cultures, which are interpreted by clinicians as a manifestation of a clinically-relevant dysphoric disorder. They also transliterated the expressions of DSM IV criteria of main dysphoric disorders in their cultures.Results: In many non-western cultures the symptoms and constructs that are interpreted and treated as dysphoric disorders are mostly somatic and are different from the Western-centered DSM or ICD systems. In many cases the DSM and ICD criteria of depression and anxieties are not even acknowledged by patients.Limitations: the descriptive approach reported here is a preliminary step which involved local but Westernized clinicians-investigators following a biomedical thinking. It should be followed by a more systematic-comprehensive surveys in each culture.Conclusions: Westernized concepts and constructs of mental order and disorders are not necessarily universally applicable. Culturallysensitive phenomena, treatments and treatment responses may be diversified. Attempts at their cross-cultural harmonization should take into consideration complex interactional multi-dimensional processes. (c) 2006 Elsevier B.V. All rights reserved.

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Actinobacillus actinomycetemcomitans plays a major role in the pathogenesis of aggressive periodontitis. Lipopolysaccharide (LPS) derived from A. actinomycetemcomitans is a key factor in inflammatory cytokine generation within periodontal tissues. In this study, we identify major mitogen-activated protein kinase (MAPK) signaling pathways induced by A. actinomycetemcomitans LPS, Escherichia coli LPS and interleukin-1 beta (IL-1 beta) in a murine periodontal ligament (mPDL) fibroblast cell line. Immunoblot analysis was used to assess the phosphorylated forms of p38, extracellular-regulated kinase (ERK) and c-jun N-terminal kinase (JNK) MAPK following stimulation with A. actinomycetemcomitans LPS, E. coli LPS and IL-1 beta. IL-6 mRNA induction was detected via reverse transcription-polymerase chain reaction, while protein levels were quantified via enzyme-linked immunosorbent assays (ELISA). We utilized biochemical inhibitors of p38, ERK and JNK MAPK to identify the MAPK signaling pathways needed for IL-6 expression. Additional use of stable mPDL cell lines containing dominant negative mutant constructs of MAPK kinase-3 and -6 (MKK-3/6) and p38 null mutant mouse embryonic fibroblast (MEF) cells were used to substantiate the biochemical inhibitor data. Blocking p38 MAPK with SB203580 reduced the induction of IL-6 mRNA by A. actinomycetemcomitans LPS, E. coli LPS and IL-1 beta by > 70%, > 95% and similar to 60%, respectively. IL-6 ELISA indicated that blocking p38 MAPK reduced the IL-6 protein levels induced by A. actinomycetemcomitans LPS, E. coli LPS and IL-1 beta by similar to 60%, similar to 50% and similar to 70%, respectively. All MAPK inhibitors significantly reduced the IL-6 protein levels induced by A. actinomycetemcomitans LPS, E. coli LPS and IL-1 beta whereas only p38 inhibitors consistently reduced the A. actinomycetemcomitans LPS, E. coli LPS and IL-1 beta induction of IL-6 mRNA steady-state levels. The contribution of p38 MAPK LPS-induced IL-6 expression was confirmed using MKK-3/6 dominant negative stable mPDL cell lines. Wild-type and p38 alpha(-/-) MEF cells provided additional evidence to support the role of p38 alpha MAPK in A. actinomycetemcomitans LPS-stimulated IL-6. Our results indicate that induction of IL-6 by E. coli LPS, IL-1 beta and A. actinomycetemcomitans LPS requires signaling through MKK-3-p38 alpha ERK, JNK and p38 MAPK in mPDL cells.

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Coupled bone turnover is directed by the expression of receptor-activated NF-kappa B ligand (RANKL) and its decoy receptor, osteoprotegerin (OPG). Proinflammatory cytokines, such as interleukin-1 beta (IL-1 beta) and tumor necrosis factor-alpha (TNF-alpha) induce RANKL expression in bone marrow stromal cells. Here, we report that IL-1 beta and TNF-alpha-induced RANKL requires p38 mitogen-activating protein kinase (MAPK) pathway activation for maximal expression. Real-time PCR was used to assess the p38 contribution toward IL-1 beta and TNF-alpha-induced RANKL mRNA expression. Steady-state RANKL RNA levels were increased approximately 17-fold by IL-1 beta treatment and subsequently reduced similar to 70%-90% when p38 MAPK was inhibited with SB203580. RANKL mRNA stability data indicated that p38 MAPK did not alter the rate of mRNA decay in IL-1 beta-induced cells. Using a RANKL-luciferase cell line receptor containing a 120-kB segment of the 5' flanking region of the RANKL gene, reporter expression was stimulated 4-5-fold by IL-1 beta or TNF-alpha treatment. IL-1 beta-induced RANKL reporter expression was completely blocked with specific p38 inhibitors as well as dominant negative mutant constructs of MAPK kinase-3 and -6. In addition, blocking p38 signaling in bone marrow stromal cells partially inhibited IL-1 beta and TNF-alpha-induced osteoclastogenesis in vitro. Results from these studies indicate that p38 MAPK is a major signaling pathway involved in IL-1 beta and TNF-alpha-induced RANKL expression in bone marrow stromal cells.

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Este trabalho, de natureza qualitativa e de cunho etnográfico, tem como cenário um Projeto de Línguas Estrangeiras, implementado nos Ciclos 1 e 2 do Ensino Fundamental Público de uma cidade do interior paulista. O objetivo principal do estudo é investigar as visões implícitas da avaliação proposta pelo livro didático em uso no mencionado contexto de ensino, confrontando-as com a abordagem explícita do material acerca do processo avaliatório e com as crenças dos professores sobre a avaliação em foco. Para tanto, tomamos como referencial os princípios sociointeracionistas (Vygotsky, 1978) e comunicacionais (Almeida Filho, 1993, 2005) de linguagem e aprendizagem, assim como construtos referentes às crenças sobre ensino e aprendizagem de línguas (Barcelos, 1995, 2004a, b; Silva, 2005) e também referentes à avaliação (Scaramucci, 1997, 2004) no ensino de línguas para crianças (Cameron, 2001).

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A new concept of fault detection and isolation using robust observation for systems with random noises is presented. The method selects the parameters from components that may fault during the process and constructs well conditioned robust observers, considering sensors faults. To isolate component failures via robust observation, a bank of detection observers is constructed, where each observer is only sensitive to one specified component failure while robust to all other component failures.

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Asian soybean rust is a formidable threat to soybean (Glycine max) production in many areas of the world, including the United States. Only five sources of resistance have been identified (Resistance to Phakopsora pachyrhizi1 [Rpp1], Rpp2, Rpp3, Rpp4, and Rpp5). Rpp4 was previously identified in the resistant genotype PI459025B and mapped within 2 centimorgans of Satt288 on soybean chromosome 18 (linkage group G). Using simple sequence repeat markers, we developed a bacterial artificial chromosome contig for the Rpp4 locus in the susceptible cv Williams82 (Wm82). Sequencing within this region identified three Rpp4 candidate disease resistance genes (Rpp4C1-Rpp4C3 [Wm82]) with greatest similarity to the lettuce (Lactuca sativa) RGC2 family of coiled coil-nucleotide binding site-leucine rich repeat disease resistance genes. Constructs containing regions of the Wm82 Rpp4 candidate genes were used for virus-induced gene silencing experiments to silence resistance in PI459025B, confirming that orthologous genes confer resistance. Using primers developed from conserved sequences in the Wm82 Rpp4 candidate genes, we identified five Rpp4 candidate genes (Rpp4C1-Rpp4C5 [PI459025B]) from the resistant genotype. Additional markers developed from the Wm82 Rpp4 bacterial artificial chromosome contig further defined the region containing Rpp4 and eliminated Rpp4C1 (PI459025B) and Rpp4C3 (PI459025B) as candidate genes. Sequencing of reverse transcription-polymerase chain reaction products revealed that Rpp4C4 (PI459025B) was highly expressed in the resistant genotype, while expression of the other candidate genes was nearly undetectable. These data support Rpp4C4 (PI459025B) as the single candidate gene for Rpp4-mediated resistance to Asian soybean rust.

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We address the different "personalities" of the rational number and the concept of proportionality, analyzing the possibilities for using the Mathematics Teaching and Learning through Problem-solving Method. This method is based on the principle that knowledge can be constructed through the use of problems that generate new concepts and new contents. The different meanings of rational number - rational point, quotient, fraction, ratio, and operator - are constructs that depend on mathematical theories in which they are imbedded and the situations that evoke them in problem-solving. Some data will be presented from continuing education courses for teachers, aiming to contribute to understanding regarding the different "personalities" of the rational number. In general, these "personalities" are not easily identified by teachers and students, which is the reason for the many difficulties encountered during problem-solving involving rational numbers. One of these "personalities", the ratio, provides the basis for the concept of proportionality, which is relevant because it is a unifying idea in mathematics.

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The main characteristics of 55 nests of Protopolybia exigua exigua were periodically evaluated in Ribeirão Preto, southeastern Brazil. Although the queens' life-span can eventually reach one year, nests barely thrive for six months because most of them are periodically invaded by the scavenger phorid fly, Megaselia aff. picta. Under heavy infestation the adult wasp population partially closes the nest entrance, migrates and constructs a new nest in the neighborhood. Afterwards the old-nest's entrance can be completely closed by the wasps which results in blocking the parasitoid flies inside. In the tropics, predation by ants is considered the main selective pressure which shaped most of the bionomic characteristics of Polistinae wasps. It is suggested that Megaselia can easily overcome the usual wasps defensive tactics just because they are so specialized against ants. Aerial raids, fast erratic-jerking movements, rapid and efficient oviposition behavior, etc. can eventually turn Megaselia into a very important enemy of some social wasps.

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A new concept of fault detection and isolation using robust observation for systems with random noises is presented. The method selects the parameters from components that may fault during the process and constructs well conditioned robust observers, considering sensors faults. To isolate component failures via robust observation, a bank of detection observers is constructed, where each observer is only sensitive to one specified component failure while robust to all other component failures.

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The state observers can reconstruct and monitor unmeasurable states. A new concept of fault detection and isolation using state observers is presented. The method selects the parameters from components that may fail during the process and constructs optimized robust observers. To isolate component failures via robust observation, a bank of detection observers is organized, in which each observer is only sensitive to one specified component failure while robust to all other component failures. This paper analyzes the performance of transient and steady-state behavior of the state observer.

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This article presents a short study about the literary work of Brazilian writer Caio Fernando Abreu (1946-1996). In order to study his formal and thematic choices, we established criteria based on concepts like motif and plot (dramatic situation), as developed by Tomachevski (1976). A specific articulation between motives and dramatic situations establishes four nuclei of Abreu's literary production, which is characterized by a literary experimentation with these elements and it constructs an organic work that makes itself and/or is made in a dialogic way (Bakhtin, 1981). Firstly, the main characteristics of the four nuclei and the books that embody them will be presented. Then, we will describe how the motives and dramatic situations are re-elaborated from one nucleus to another. Not only does each nucleus features some new elements, but also contains remainders of previous ones. As such, each of Abreu's works can be considered as both the same and unique.

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The importance of thrombosis and anticoagulation in clinical practice is rooted firmly in several fundamental constructs that can be applied both broadly and globally. Awareness and the appropriate use of anticoagulant therapy remain the keys to prevention and treatment. However, to assure maximal efficacy and safety, the clinician must, according to the available evidence, choose the right drug, at the right dose, for the right patient, under the right indication, and for the right duration of time. The first International Symposium of Thrombosis and Anticoagulation in Internal Medicine was a scientific program developed by clinicians for clinicians. The primary objective of the meeting was to educate, motivate and inspire internists, cardiologists and hematologists by convening national and international visionaries, thought-leaders and dedicated clinician-scientists in Sao Paulo, Brazil. This article is a focused summary of the symposium proceedings. © Springer Science+Business Media, LLC 2009.

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Background: Protein-calorie malnutrition (PCM) is the most common type of malnutrition. PCM leads to immunodeficiency and consequent increased susceptibility to infectious agents. In addition, responses to prophylactic vaccines depend on nutritional status. This study aims to evaluate the ability of undernourished mice to mount an immune response to a genetic vaccine (pVAXhsp65) against tuberculosis, containing the gene coding for the heat shock protein 65 from mycobacteria. Methods: Young adult female BALB/c mice were fed ad libitum or with 80% of the amount of food consumed by a normal diet group. We initially characterized a mice model of dietary restriction by determining body and spleen weights, hematological parameters and histopathological changes in lymphoid organs. The ability of splenic cells to produce IFN-gamma and IL-4 upon in vitro stimulation with LPS or S. aureus and the serum titer of specific IgG1 and IgG2a anti-hsp65 antibodies after intramuscular immunization with pVAXhsp65 was then tested. Results: Dietary restriction significantly decreased body and spleen weights and also the total lymphocyte count in blood. This restriction also determined a striking atrophy in lymphoid organs as spleen, thymus and lymphoid tissue associated with the small intestine. Specific antibodies were not detected in mice submitted to dietary restriction whereas the well nourished animals produced significant levels of both, IgG1 and IgG2a anti-hsp65. Conclusion: 20% restriction in food intake deeply compromised humoral immunity induced by a genetic vaccine, alerting, therefore, for the relevance of the nutritional condition in vaccination programs based on these kinds of constructs. © 2009 Ishikawa et al; licensee BioMed Central Ltd.

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Purpose: This paper aims to perform an empirical investigation about the constructs and indicators of the supply chain management practices framework. Design/methodology/approach: The measuring framework proposed is based on a survey that was carried out on 107 Brazilian companies. Statistical techniques were employed to verify, validate, and test the reliability of the constructs and their indicators. To validate this framework principal component analysis and structural equation modeling techniques were used. Findings: In general, previous studies suggest six constructs for measuring the supply chain management practices framework. However, in this study a framework was achieved with four constructs of supply chain management practices, namely, supply chain (SC) integration for production planning and control (PPC) support, information sharing about products and targeting strategies, strategic relationship with customer and supplier, and support customer order. This framework has adequate levels of validity and reliability. Research limitations/implications: The main limitation of this study was that only a small sample of companies in a single sector and country were surveyed, and therefore there needs to be further research considering the special conditions in other countries. Originality/value: This study investigated statistically set indicators to discuss the topic supply chain management practices. The framework obtained has good quality of validity and reliability indicators. Thus, an alternative framework has been added to measure supply chain management practices, which is currently a popular topic in the supply chain mainstream literature. Both defined constructs and the validated indicators can be used in other studies on supply chain management. © Emerald Group Publishing Limited.