Existence of multiple solutions for finite difference approximations to second-order boundary value problems

Error condition detected We consider discrete two-point boundary value problems of the form D-2 y(k+1) = f (kh, y(k), D y(k)), for k = 1,...,n - 1, (0,0) = G((y(0),y(n));(Dy-1,Dy-n)), where Dy-k = (y(k) - Yk-I)/h and h = 1/n. This arises as a finite difference approximation to y" = f(x,y,y'), x is an element of [0,1], (0,0) = G((y(0),y(1));(y'(0),y'(1))). We assume that f and G = (g(0), g(1)) are continuous and fully nonlinear, that there exist pairs of strict lower and strict upper solutions for the continuous problem, and that f and G satisfy additional assumptions that are known to yield a priori bounds on, and to guarantee the existence of solutions of the continuous problem. Under these assumptions we show that there are at least three distinct solutions of the discrete approximation which approximate solutions to the continuous problem as the grid size, h, goes to 0. (C) 2003 Elsevier Science Ltd. All rights reserved.

Estoque e frações de carbono orgânico e fertilidade de solo sob floresta, agricultura e pecuária

O entendimento dos fenômenos de trocas de carbono entre solo, água e atmosfera é fundamental para o manejo e a conservação de ecossistemas naturais e agrários. Os objetivos deste trabalho foram determinar os estoques de carbono orgânico, as suas frações e a fertilidade do solo sob os manejos florestal (mata remanescente), integrando agricultura e florestas (eucalipto e cutieira) e em monoculturas agrícola (sob pivô central) e pastoril (braquiária), na Bacia do Rio Paraopeba, em Florestal-MG, visando a propor indicadores para a avaliação de manejo e conservação do solo. As amostras de solo foram coletadas em remanescente de mata nativa (Mata), em área agrícola cultivada sob irrigação de pivô central (Pivô) e em áreas de cultivo de Corymbia citriodora (Corymbia citriodora (Hook.) K.D. Hill & L.A.S. Johnson) (Eucalipto), Joannesia princeps Vell. (Cutieira) e Brachiaria decumbens Stapf (Pasto), nas profundidades de 0 a 20 e 20 a 40 cm. Os resultados indicaram que as estimativas dos estoques e de estabilidade do carbono orgânico aumentaram com a fertilidade do solo. Os sistemas florestais, principalmente a Mata, acompanhada pelos sistemas de manejo do solo integrando florestas à pastagem, em comparação com o cultivo contínuo ou a monocultura de pastagem, apresentaram estoques de carbono maiores, mais estáveis e menos solúveis, com formas mais aromáticas e hidrofóbicas (maior relação AH/AF), indicando menor potencial de lixiviação de carbono para o sistema aquático adjacente.

Chromatographic behaviour of monoclonal antibodies against wild-type amidase from Pseudomonasaeruginosa on immobilized metal chelates

The aim of this work was to devise a one-step purification procedure for monoclonal antibodies (MAbs) of IgG class by immobilized metal affinity chromatography (IMAC). Therefore, several stationary phases were prepared containing immobilized metal chelates in order to study the chromatographic behaviour of MAbs against wild-type amidase from Pseudomonas aeruginosa. Such MAbs adsorbed to Cu(II), Ni(II), Zn(II) and Co(II)-IDA agarose columns. The increase in ligand concentration and the use of longer spacer arms and higher pH values resulted in higher adsorption of MAbs into immobilized metal chelates. The dynamic binding capacity and the maximum binding capacity were 1.33 +/- 0.015 and 3.214 +/- 0.021 mg IgG/mL of sedimented commercial matrix, respectively. A K(D) of 4.53 x 10(-7) M was obtained from batch isotherm measurements. The combination of tailor-made stationary phases of IMAC and the correct selection of adsorption conditions permitted a one-step purification procedure to be devised for MAbs of IgG class. Culture supernatants containing MAbs were purified by IMAC on commercial-Zn(II) and EPI-30-IDA-Zn(II) Sepharose 6B columns and by affinity chromatography on Protein A-Sepharose CL-4B. This MAb preparation revealed on SDS-PAGE two protein bands with M(r) of 50 and 22 kDa corresponding to the heavy and light chains, respectively. Copyright (C) 2011 John Wiley & Sons, Ltd.

Predictivity Strength of the Spatial Variability of Phenanthrene Sorption Across Two Sandy Loam Fields

Sorption is commonly agreed to be the major process underlying the transport and fate of polycyclic aromatic hydrocarbons (PAHs) in soils. However, there is still a scarcity of studies focusing on spatial variability at the field scale in particular. In order to investigate the variation in the field of phenanthrene sorption, bulk topsoil samples were taken in a 15 × 15-m grid from the plough layer in two sandy loam fields with different texture and organic carbon (OC) contents (140 samples in total). Batch experiments were performed using the adsorption method. Values for the partition coefficient K d (L kg−1) and the organic carbon partition coefficient K OC (L kg−1) agreed with the most frequently used models for PAH partitioning, as OC revealed a higher affinity for sorption. More complex models using different OC compartments, such as non-complexed organic carbon (NCOC) and complexed organic carbon (COC) separately, performed better than single K OC models, particularly for a subset including samples with Dexter n < 10 and OC <0.04 kg kg−1. The selected threshold revealed that K OC-based models proved to be applicable for more organic fields, while two-component models proved to be more accurate for the prediction of K d and retardation factor (R) for less organic soils. Moreover, OC did not fully reflect the changes in phenanthrene retardation in the field with lower OC content (Faardrup). Bulk density and available water content influenced the phenanthrene transport mechanism phenomenon.

Providing Adaptation for an Adaptive Hypermedia System

O ensino à distância cresceu consideravelmente nos últimos anos e a tendência é para que continue a crescer em anos vindouros. No entanto, enquanto que a maioria das plataformas de ensino à distância utilizam a mesma abordagem de ensino para todos os utilizadores, os estudantes que as usam são na realidade pessoas de diferentes culturas, locais, idades e géneros, e que possuem diferentes níveis de educação. Ao contrário do ensino à distância tradicional, os sistemas de hipermédia adaptativa educacional adaptam interface, apresentação de conteúdos e navegação, entre outros, às características, necessidades e interesses específicos de diferentes utilizadores. Apesar da investigação na área de sistemas de hipermédia adaptativa já estar bastante desenvolvida, é necessário efetuar mais desenvolvimento e experimentação de modo a determinar quais são os aspetos mais eficazes destes sistemas e avaliar o seu sucesso. A Plataforma de Aprendizagem Colaborativa da Matemática (PCMAT) é um sistema de hipermédia adaptativa educacional com uma abordagem construtivista, que foi desenvolvido com o objetivo de contribuir para a investigação na área de sistemas de hipermédia adaptativa. A plataforma avalia o conhecimento do utilizador e apresenta conteúdos e atividades adaptadas às características e estilo de aprendizagem dominante de estudantes de matemática do segundo ciclo. O desenvolvimento do PCMAT tem também o propósito de auxiliar os alunos Portugueses com a aprendizagem da matemática. De acordo com o estudo PISA 2012 da OCDE [OECD, 2014], o desempenho dos alunos Portugueses na área da matemática melhorou em relação à edição anterior do estudo, mas os resultados obtidos permanecem abaixo da média da OCDE. Por este motivo, uma das finalidades deste projeto é desenvolver um sistema de hipermédia adaptativa que, ao adequar o ensino da matemática às necessidades específicas de cada aluno, os assista com a aquisição de conhecimento. A adaptação é efetuada pelo sistema usando a informação constante no modelo do utilizador para definir um grafo de conceitos do domínio específico. Este grafo é adaptado do modelo do domínio e utilizado para dar resposta às necessidades particulares de cada aluno. Embora a trajetória inicial seja definida pelo professor, o percurso percorrido no grafo por cada aluno é determinado pela sua interação com o sistema, usando para o efeito a representação do conhecimento do aluno e outras características disponíveis no modelo do utilizador, assim como avaliação progressiva. A adaptação é conseguida através de alterações na apresentação de conteúdos e na estrutura e anotações das hiperligações. A apresentação de conteúdos é alterada mostrando ou ocultando cada um dos vários fragmentos que compõe as páginas dum curso. Estes fragmentos são compostos por diferentes objetos de aprendizagem, tais como exercícios, figuras, diagramas, etc. As mudanças efetuadas na estrutura e anotações das hiperligações têm o objetivo de guiar o estudante, apontando-o na direção do conhecimento mais relevante e mantendo-o afastado de informação inadequada. A escolha de objectos de aprendizagem adequados às características particulares de cada aluno é um aspecto essencial do modelo de adaptação do PCMAT. A plataforma inclui para esse propósito um módulo responsável pela recomendação de objectos de aprendizagem, e um módulo para a pesquisa e recuperação dos mesmos. O módulo de recomendação utiliza lógica Fuzzy para converter determinados atributos do aluno num conjunto de parâmetros que caracterizam o objecto de aprendizagem que idealmente deveria ser apresentado ao aluno. Uma vez que o objecto “ideal” poderá não existir no repositório de objectos de aprendizagem do sistema, esses parâmetros são utilizados pelo módulo de pesquisa e recuperação para procurar e devolver ao módulo de recomendação uma lista com os objectos que mais se assemelham ao objecto “ideal”. A pesquisa é feita numa árvore k-d usando o algoritmo k-vizinhos mais próximos. O modelo de recomendação utiliza a lista devolvida pelo módulo de pesquisa e recuperação para seleccionar o objecto de aprendizagem mais apropriado para o aluno e processa-o para inclusão numa das páginas Web do curso. O presente documento descreve o trabalho desenvolvido no âmbito do projeto PCMAT (PTDS/CED/108339/2008), dando relevância à adaptação de conteúdos.

Gd(III) chelates as NMR probes of protein-protein interactions. Case study: rubredoxin and cytochrome c3

Inorganic Chemistry 50(21):10600-7

A copper protein and a cytochrome bind at the same site on bacterial cytochrome c peroxidase

Biochemistry, 2004, 43 (46), pp 14566–14576 DOI: 10.1021/bi0485833

Polyanalgesic Consensus Conference--2012: recommendations on trialing for intrathecal (intraspinal) drug delivery: report of an interdisciplinary expert panel.

INTRODUCTION: Trialing for intrathecal pump placement is an essential part of the decision-making process in placing a permanent device. In both the United States and the international community, the proper method for trialing is ill defined. METHODS: The Polyanalgesic Consensus Conference (PACC) is a group of well-published experienced practitioners who meet to update the state of care for intrathecal therapies on the basis of current knowledge in the literature and clinical experience. Anexhaustive search is performed to create a base of information that the panel considers when making recommendations for best clinical practices. This literature, coupled with clinical experience, is the basis for recommendations and for identification of gaps in the base of knowledge regarding trialing for intrathecal pump placement. RESULTS: The panel has made recommendations for the proper methods of trialing for long-term intrathecal drug delivery. CONCLUSION: The use of intrathecal drug delivery is an important part of the treatment algorithm for moderate to severe chronic pain. It has become common practice to perform a temporary neuroaxial infusion before permanent device implantation. On the basis of current knowledge, the PACC has developed recommendations to improve care. The need to update these recommendations will be very important as new literature is published.

Identification and analysis of functional elements in 1% of the human genome by the ENCODE pilot project.

We report the generation and analysis of functional data from multiple, diverse experiments performed on a targeted 1% of the human genome as part of the pilot phase of the ENCODE Project. These data have been further integrated and augmented by a number of evolutionary and computational analyses. Together, our results advance the collective knowledge about human genome function in several major areas. First, our studies provide convincing evidence that the genome is pervasively transcribed, such that the majority of its bases can be found in primary transcripts, including non-protein-coding transcripts, and those that extensively overlap one another. Second, systematic examination of transcriptional regulation has yielded new understanding about transcription start sites, including their relationship to specific regulatory sequences and features of chromatin accessibility and histone modification. Third, a more sophisticated view of chromatin structure has emerged, including its inter-relationship with DNA replication and transcriptional regulation. Finally, integration of these new sources of information, in particular with respect to mammalian evolution based on inter- and intra-species sequence comparisons, has yielded new mechanistic and evolutionary insights concerning the functional landscape of the human genome. Together, these studies are defining a path for pursuit of a more comprehensive characterization of human genome function.

Prevalence of and potential influencing factors for alcohol dependence in europe.

Alcohol use disorders (AUDs), and alcohol dependence (AD) in particular, are prevalent and associated with a large burden of disability and mortality. The aim of this study was to estimate prevalence of AD in the European Union (EU), Iceland, Norway, and Switzerland for the year 2010, and to investigate potential influencing factors. The 1-year prevalence of AD in the EU was estimated at 3.4% among people 18-64 years of age in Europe (women 1.7%, men 5.2%), resulting in close to 11 million affected people. Taking into account all people of all ages, AD, abuse and harmful use resulted in an estimate of 23 million affected people. Prevalence of AD varied widely between European countries, and was significantly impacted by drinking cultures and social norms. Correlations with level of drinking and other drinking variables and with major known outcomes of heavy drinking, such as liver cirrhosis or injury, were moderate. These results suggest a need to rethink the definition of AUDs.

SHP-1 phosphatase activity counteracts increased T cell receptor affinity.

Anti-self/tumor T cell function can be improved by increasing TCR-peptide MHC (pMHC) affinity within physiological limits, but paradoxically further increases (K(d) < 1 μM) lead to drastic functional declines. Using human CD8(+) T cells engineered with TCRs of incremental affinity for the tumor antigen HLA-A2/NY-ESO-1, we investigated the molecular mechanisms underlying this high-affinity-associated loss of function. As compared with cells expressing TCR affinities generating optimal function (K(d) = 5 to 1 μM), those with supraphysiological affinity (K(d) = 1 μM to 15 nM) showed impaired gene expression, signaling, and surface expression of activatory/costimulatory receptors. Preferential expression of the inhibitory receptor programmed cell death-1 (PD-1) was limited to T cells with the highest TCR affinity, correlating with full functional recovery upon PD-1 ligand 1 (PD-L1) blockade. In contrast, upregulation of the Src homology 2 domain-containing phosphatase 1 (SHP-1/PTPN6) was broad, with gradually enhanced expression in CD8(+) T cells with increasing TCR affinities. Consequently, pharmacological inhibition of SHP-1 with sodium stibogluconate augmented the function of all engineered T cells, and this correlated with the TCR affinity-dependent levels of SHP-1. These data highlight an unexpected and global role of SHP-1 in regulating CD8(+) T cell activation and responsiveness and support the development of therapies inhibiting protein tyrosine phosphatases to enhance T cell-mediated immunity.

Genetic variants in novel pathways influence blood pressure and cardiovascular disease risk.

Blood pressure is a heritable trait influenced by several biological pathways and responsive to environmental stimuli. Over one billion people worldwide have hypertension (≥140 mm Hg systolic blood pressure or  ≥90 mm Hg diastolic blood pressure). Even small increments in blood pressure are associated with an increased risk of cardiovascular events. This genome-wide association study of systolic and diastolic blood pressure, which used a multi-stage design in 200,000 individuals of European descent, identified sixteen novel loci: six of these loci contain genes previously known or suspected to regulate blood pressure (GUCY1A3-GUCY1B3, NPR3-C5orf23, ADM, FURIN-FES, GOSR2, GNAS-EDN3); the other ten provide new clues to blood pressure physiology. A genetic risk score based on 29 genome-wide significant variants was associated with hypertension, left ventricular wall thickness, stroke and coronary artery disease, but not kidney disease or kidney function. We also observed associations with blood pressure in East Asian, South Asian and African ancestry individuals. Our findings provide new insights into the genetics and biology of blood pressure, and suggest potential novel therapeutic pathways for cardiovascular disease prevention.

Prostaglandin transporter mutations cause pachydermoperiostosis with myelofibrosis.

Pachydermoperiostosis, or primary hypertrophic osteoarthropathy (PHO), is an inherited multisystem disorder, whose features closely mimic the reactive osteoarthropathy that commonly accompanies neoplastic and inflammatory pathologies. We previously described deficiency of the prostaglandin-degrading enzyme 15-hydroxyprostaglandin dehydrogenase (HPGD) as a cause of this condition, implicating elevated circulating prostaglandin E(2) (PGE(2) ) as causative of PHO, and perhaps also as the principal mediator of secondary HO. However, PHO is genetically heterogeneous. Here, we use whole-exome sequencing to identify recessive mutations of the prostaglandin transporter SLCO2A1, in individuals lacking HPGD mutations. We performed exome sequencing of four probands with severe PHO, followed by conventional mutation analysis of SLCO2A1 in nine others. Biallelic SLCO2A1 mutations were identified in 12 of the 13 families. Affected individuals had elevated urinary PGE(2) , but unlike HPGD-deficient patients, also excreted considerable quantities of the PGE(2) metabolite, PGE-M. Clinical differences between the two groups were also identified, notably that SLCO2A1-deficient individuals have a high frequency of severe anemia due to myelofibrosis. These findings reinforce the key role of systemic or local prostaglandin excess as the stimulus to HO. They also suggest that the induction or maintenance of hematopoietic stem cells by prostaglandin may depend upon transporter activity. Hum Mutat 33:1175-1181, 2012. © 2012 Wiley Periodicals, Inc.