Targeting of EBNA1 for rapid intracellular degradation overrides the inhibitory effects of the Gly-Ala repeat domain and restores CD8+T cell recognition

Epstein-Barr virus (EBV)-encoded nuclear antigen 1 (EBNA1) includes a unique glycine-alanine repeat domain that inhibits the endogenous presentation of cytotoxic T lymphocyte (CTL) epitopes through the class I pathway by blocking proteasome-dependent degradation of this antigen. This immune evasion mechanism has been implicated in the pathogenesis of EBV-associated diseases. Here, we show that cotranslational ubiquitination combined with N-end rule targeting enhances the intracellular degradation of EBNA1, thus resulting in a dramatic reduction in the half-life of the antigen. Using DNA expression vectors encoding different forms of ubiquitinated EBNA1 for in vivo studies revealed that this rapid degradation, remarkably, leads to induction of a very strong CTL response to an EBNA1-specific CTL epitope. Furthermore, this targeting also restored the endogenous processing of HLA class I-restricted CTL epitopes within EBNA1 for immune recognition by human EBV-specific CTLs. These observations provide, for the first time, evidence that the glycine-alanine repeat-mediated proteasomal block on EBNA1 can be reversed by specifically targeting this antigen for rapid degradation resulting in enhanced CD8+ T cell-mediated recognition in vitro and in vivo.

Adoptive transfer of autologous Epstein-Barr virus-specific cytotoxic T cells for nasopharyngeal carcinoma

Tumor cells from NPC patients are regularly and latently infected with EBV. To examine whether the virus serves as target for immune intervention of the cancer, we determined levels of EBV-specific CTLp in peripheral blood from NPC patients, long-term survivors of the cancer and healthy subjects. CTLp levels of test subjects varied between 3-3,000/10(6) PBMCs. The plasma EBV burden increased when the CTLp level fell below 150, whereas the EBV burden of PBMCs was not correlated with CTLp level. Compared with healthy carriers, CTLp levels of patients were lower and varied over a wider range, between 3-1,500/10(6) PBMCs. The quantitative immune deficit was probably attributed to the tumor because, first, CTLp in survivors was restored to levels similar to those in healthy carriers after the tumor had been successfully treated. Second, the CTLp level changed as disease progressed, being lower in local disease, increased in locoregional disease and decreased again when the tumor metastasized. Based on these findings, 4 patients with advanced disease were infused with 5 x 10(7)-3 x 10(8) autologous EBV CTLs. The treatment was safe and unaccompanied by inflammatory or other complications, but whether it improved tumor control could not be discerned from the large tumor bulk. Nevertheless, the treatment regularly increased CTLp levels of patients, maintained it at higher levels for protracted periods and, in 3 patients, restored host surveillance of EBV replication, reducing the plasma EBV burden. Taken together, these results provided a rationale to further explore EBV as a target of immune intervention of NPC. (C) 2001 Wiley-Liss, Inc.

The immunology of Epstein-Barr virus infection

Epstein-Barr virus is a classic example of a persistent human virus that has caught the imagination of immunologists, virologists and oncologists because of the juxtaposition of a number of important properties. First, the ability of the virus to immortalize B lymphocytes in vitro has provided an antigen presenting cell in which all the latent antigens: of the virus are displayed and are available for systematic study. Second, the virus presents an ideal system for studying the immune parameters that maintain latency and the consequences of disturbing this cell-virus relationship. Third, this wealth of immunological background has provided a platform for elucidating the role of the immune system in protection from viral-associated malignancies of B cell and epithelial cell origin. Finally attention is now being directed towards the development of vaccine formulations which might have broad application in the control of human malignancies.

Neonatal hepatic drug elimination

After the transition from in utero to newborn life, the neonate becomes solely reliant upon its own drug clearance processes to metabolise xenobiotics. Whilst most studies of neonatal hepatic drug elimination have focussed upon in vitro expression and activities of drug-metabolising enzymes, the rapid physiological changes in the early neonatal period of life also need to be considered. There are dramatic changes in neonatal liver blood how and hepatic oxygenation due to the loss of the umbilical blood supply, the increasing portal vein blood flow, and the gradual closure of the ductus venosus shunt during the first week of life. These changes which may well affect the capacity of neonatal hepatic drug metabolism. The hepatic expression of cytochromes P450 1A2, 2C, 2D6, 2E1 and 3A4 develop at different rates in the postnatal period, whilst 3A7 expression diminishes. Hepatic glucuronidation in the human neonate is relatively immature at birth, which contrasts with the considerably more mature neonatal hepatic sulfation activity. Limited in vivo studies show that the human neonate can significantly metabolise xenobiotics but clearance is considerably less compared with the older infant and adult. The neonatal population included in pharmacological studies is highly heterogeneous with respect to age, body weight, ductus venosus closure and disease processes, making it difficult to interpret data arising from human neonatal studies. Studies in the perfused foetal and neonatal sheep liver have demonstrated how the oxidative and conjugative hepatic elimination of drugs by the intact organ is significantly increased during the first week of life, highlighting that future studies will need to consider the profound physiological changes that may influence neonatal hepatic drug elimination shortly after birth.

Thiopalmitoylation of myelin proteolipid protein epitopes enhances immunogenicity and encephalitogenicity

Proteolipid protein (PLP) is the most abundant protein of CNS myelin, and is posttranslationally acylated by covalent attachment of long chain fatty acids to cysteine residues via a thioester linkage. Two of the acylation sites are within epitopes of PLP that are encephalitogenic in SJL/J mice (PLP104-117 and PLP139-151) and against which increased immune responses have been detected in some multiple sclerosis patients. It is known that attachment of certain types of lipid side chains to peptides can result in their enhanced immunogenicity. The aim of this study was to determine whether thioacylated PLP peptides, as occur in the native protein, are more immunogenic than their nonacylated counterparts, and whether thioacylation influences the development of autoreactivity and experimental autoimmune encephalomyelitis. The results show that in comparison with nonacylated peptides, thioacylated PLP lipopeptides can induce greater T cell and Ab responses to both the acylated and nonacylated peptides. They also enhanced the development and chronicity of experimental autoimmune encephalomyelitis. Synthetic peptides in which the fatty acid was attached via an amide linkage at the N terminus were not encephalitogenic, and they induced greater proportions of CD8(+) cells in initial in vitro stimulation. Therefore, the lability and the site of the linkage between the peptide and fatty acid may be important for induction of encephalitogenic CD4(+) T cells. These results suggest that immune responses induced by endogenous thioacylated lipopeptides may contribute to the immunopathogenesis of chronic experimental demyelinating diseases and multiple sclerosis.

Method for determining the shear stress in cylindrical systems

We develop a method for determining the elements of the pressure tensor at a radius r in a cylindrically symmetric system, analogous to the so-called method of planes used in planar systems [B. D. Todd, Denis J. Evans, and Peter J. Daivis, Phys. Rev. E 52, 1627 (1995)]. We demonstrate its application in determining the radial shear stress dependence during molecular dynamics simulations of the forced flow of methane in cylindrical silica mesopores. Such expressions are useful for the examination of constitutive relations in the context of transport in confined systems.

A hist??ria de Andrea: como manter uma postura profissional quando n??o se sabe como agir?

Andrea, apesar de bem jovem, foi promovida ?? chefia de uma equipe com profissionais experientes e qualificados, mas as dificuldades come??aram logo. Sem saber lidar com isso, ela come??ou a se esquivar de situa????es dif??ceis, deixando a solu????o para a sua experiente equipe. Apesar disso, Andrea continuava com uma boa imagem perante seus superiores. Ao final do caso, prop??e-se um exerc??cio de avalia????o sobre as habilidades de Andrea

A hist??ria de Henrique: quando as habilidades de l??der s??o testadas ap??s a ascens??o profissional

Henrique, funcion??rio antigo de um importante programa governamental, ?? promovido ao mais alto cargo do programa. Devido a algumas posi????es e a????es ortodoxas, colegas e superiores come??aram a questionar o estilo e as iniciativas de Henrique. Ele, por sua vez, ao identificar as cr??ticas, voltou-se para fora da institui????o, transformando-se em ???embaixador do programa???, com viagens semanais e cada vez mais ausente. Seus subordinados tamb??m come??aram a se queixar e o Ministro finalmente mostrou insatisfa????o a respeito. Ao final do caso, prop??e-se um exerc??cio de avalia????o sobre as habilidades de Henrique

A hist??ria de Ricardo: raz??es pretensamente boas podem render falta de motiva????o

Ap??s torna-se Diretor-Geral da empresa, Ricardo desejava transform??-la num centro de excel??ncia. Para tanto, empenhou-se em focar em resultados financeiros, deixando de lado a criatividade e a autonomia profissional da equipe, o que gerou descontentamento e demiss??es. Ao final do caso, prop??e-se um exerc??cio de avalia????o sobre as habilidades de Ricardo

Disciplina 4.6: gest??o, avalia????o e revis??o do PPA

Ciclo de gest??o do PPA; gest??o integrada do plano; a responsabiliza????o por resultados. Modelo e Gest??o do PPA 2012-2015: os atores, arranjos para a gest??o dos programas e Sistema de Monitoramento e Avalia????o. Monitoramento de Programas: conceitos gerais utilizados no governo federal, tipos de monitoramento, informa????es gerenciais e gest??o de restri????es. Instrumentos de monitoramento e sistemas da Uni??o. Monitoramento intensivo. Avalia????o de Programas: conceito e tipos de avalia????o. Avalia????o do PPA. Revis??o do Plano

O plano plurianual municipal no sistema de planejamento e or??amento brasileiro

A partir da perspectiva do New Public Financial Management, o artigo traz os aspectos t??cnicos e pol??ticos que permearam o desenvolvimento do sistema de planejamento e or??amento brasileiro. Considerando os condicionantes dos momentos hist??ricos que interferiram em seu desenvolvimento, s??o identificadas caracter??sticas pol??ticas, t??cnicas e metodol??gicas incidentes no controle financeiro, nas melhorias administrativas, no planejamento e, finalmente, na tend??ncia participativa decorrente da amplia????o de espa??os democr??ticos. Em seguida, s??o apresentados o tratamento constitucional dado ao Plano Plurianual e o aperfei??oamento do sistema de planejamento e or??amento trazido pela Lei de Responsabilidade Fiscal. S??o destacadas, para o caso municipal, as inova????es contidas no Estatuto da Cidade, com reflexos no Plano Plurianual Municipal. Ressalta-se a import??ncia do processo de planejamento e or??amento na execu????o das pol??ticas p??blicas, enfatizando as dificuldades ainda existentes quanto ?? regula????o do sistema de planejamento e or??amento para permitir o imbricamento dessas pol??ticas entre as tr??s esferas federativas.

Dacriocistorrinostomia endonasal em crianças

OBJETIVO: Nosso objetivo foi verificar se os resultados com a dacriocistorrinostomia endonasal endoscópica em crianças com obstrução do canal nasolacrimal permitem considerá-la uma alternativa de tratamento aceitável para crianças. FORMA DE ESTUDO: clínico com coorte transversal. MATERIAL E MÉTODO: Vinte e sete dacriocistorrinostomias endonasais endoscópicas foram realizadas em crianças de 2 a 12 anos com obstrução do canal nasolacrimal. Todas as crianças haviam sido tratadas por sondagem lacrimal, sem resultado. A técnica utilizou uncinectomia e uma abertura do saco lacrimal pequena. O seguimento foi de 3 meses. RESULTADOS: Bom resultado foi verificado após 21 cirurgias (77,8% de sucesso). A única complicação verificada foi prolapso da intubação com silicone em 1 caso. CONCLUSÃO: Nossos resultados permitem confirmar a dacriocistorrinostomia endonasal endoscópica como alternativa válida e segura para o tratamento de crianças com obstrução nasolacrimal resistente a sondagens.

Os sites dos ministérios de relações exteriores no mundo e a História

A Internet é aqui considerada uma fonte alternativa do estudo da história, sobretudo história das relações diplomáticas. São analisados os sites dos ministérios das relações exteriores pelo mundo: alguns atribuem grande importância à história, com detalhada visão retrospectiva (Brasil, França, México, Rússia, Alemanha, Estados Unidos), outros têm abordagem seletiva ou restrita a determinados períodos (Espanha, Argentina, Itália, Bélgica, Portugal, China, Japão), uns se voltam para o passado, somente com informações históricas que sejam indispensáveis à imagem nacional (Suíça) e outros mostram sua história voltada para o futuro (Grã-Bretanha).

Real-time 3D interactive segmentation of echocardiographic data through user-based deformation of B-spline explicit active surfaces

Image segmentation is an ubiquitous task in medical image analysis, which is required to estimate morphological or functional properties of given anatomical targets. While automatic processing is highly desirable, image segmentation remains to date a supervised process in daily clinical practice. Indeed, challenging data often requires user interaction to capture the required level of anatomical detail. To optimize the analysis of 3D images, the user should be able to efficiently interact with the result of any segmentation algorithm to correct any possible disagreement. Building on a previously developed real-time 3D segmentation algorithm, we propose in the present work an extension towards an interactive application where user information can be used online to steer the segmentation result. This enables a synergistic collaboration between the operator and the underlying segmentation algorithm, thus contributing to higher segmentation accuracy, while keeping total analysis time competitive. To this end, we formalize the user interaction paradigm using a geometrical approach, where the user input is mapped to a non-cartesian space while this information is used to drive the boundary towards the position provided by the user. Additionally, we propose a shape regularization term which improves the interaction with the segmented surface, thereby making the interactive segmentation process less cumbersome. The resulting algorithm offers competitive performance both in terms of segmentation accuracy, as well as in terms of total analysis time. This contributes to a more efficient use of the existing segmentation tools in daily clinical practice. Furthermore, it compares favorably to state-of-the-art interactive segmentation software based on a 3D livewire-based algorithm.

Disponibilidade de nitrogênio de fontes minerais e orgânicas aplicadas em um Argissolo cultivado com videira

As videiras em produção tendem a aproveitar pequena quantidade do N, derivado do fertilizante mineral, mesmo cultivadas em solos com textura arenosa e baixo teor de matéria orgânica, o que confere a esses solos baixa capacidade de fornecimento de nitrogênio (N). O objetivo deste trabalho foi avaliar, por meio de incubação, a disponibilidade de nitrogênio (N) em solo com histórico de cultivo com videiras e submetido à aplicação de fontes minerais e orgânicas de nutrientes. Foram coletadas amostras de um Argissolo Vermelho, em um vinhedo e em uma área sob campo natural, no município de Rosário do Sul (RS). Após o preparo do solo, foram montados os seguintes tratamentos: solo do campo nativo; solo do vinhedo; solo do vinhedo + composto orgânico; solo do vinhedo + ureia revestida com polímeros e solo do vinhedo + ureia. Os solos foram incubados por 141 dias, e aos 0, 38, 70, 102 e 141 dias de incubação, foram determinados os teores de N-NH4+ e N-NO3-, e calculados os teores de N mineral, mineralização líquida, N mineralizado em relação ao total e o N mineral acumulado. No solo de vinhedo submetido à aplicação de ureia revestida com polímeros e de ureia, a maior mineralização de N aconteceu no período inicial. A liberação de N do composto orgânico tem um maior sincronismo com a necessidade da videira pelo nutriente. As reservas de N total potencialmente mineralizável do solo podem suprir a demanda da videira pelo N.