bluuebunny/crossref_metadata_2025 huggingface dataset:
A tripartição pronominal e o estatuto das proformas cê, ocê e você
Authors: Petersen Carol
Published: September 2009

Este artigo mostra que os argumentos apresentados em Ramos (1997) e Vitral (1996) ao defenderem a proposta de classificação da proforma cê como um clítico não conseguem dar conta do comportamento particular desta proforma no sistema pronominal do português brasileiro. Para resolver o problema, realizo uma reanálise destes argumentos em função da teoria da tripartição pronominal de Cardinaletti e Starke (1999). De acordo com esta perspectiva, as evidências apresentadas pelos autores a favor da cliticização de cê mostram apenas que cê não é um pronome forte. Ao ser identificado como um pronome fraco, pode-se então explicar o contraste entre esta proforma e ocê e você, proformas que possuem uma variedade forte e outra fraca.
Apoptosis in pulp elimination during physiological root resorption in human primary teeth
Authors: Rodrigues Luciana Villela , Vasconcelos Anilton César , Campos Pedro Alves , Brant Juliana Massote Caldeira
Published: September 2009

Pulp samples of 50 healthy human teeth with indication for extraction were examined to evaluate the role of apoptosis in pulp elimination during physiological root resorption. Two groups were formed: a test group (n=30) composed of pulp samples of primary teeth with physiological root resorption and a control group (n=20) composed of pulp samples of permanent maxillary third molars. Morphological evidence of apoptosis as well as in situ detection of cellular DNA fragmentation by TUNEL assay and detection of internucleosomal pattern of fragmentation of the genomic DNA by electrophoresis were observed. The apoptotic index of the primary tooth group was significantly higher than that of the permanent tooth group (51.01 ± 0.52 versus 25.32 ± 0.68) (p<0.001). TUNEL reaction showed intense and diffuse labeling in the pulp samples of primary teeth, which were discrete in the controls. Intense DNA internucleosomal fragmentation, a specific pattern for apoptosis, was observed in primary tooth pulps DNA by electrophoresis, in the permanent tooth pulps this pattern fragmentation of the genomic DNA for apoptosis were not present. These results seem to indicate a role of apoptosis in pulp elimination during the physiological root resorption of human primary teeth.
"Isso é política, meu!" socialização militante e institucionalização dos movimentos sociais
Authors: Moreno Rosangela Carrilo , Almeida Ana Maria F.
Published: September 2009

Como se cria uma associação militante? Que elementos e/ou processos são necessários para que indivíduos ajam como um grupo para criar uma organização destinada a atuar no espaço político? Esta questão foi discutida numa pesquisa que focalizou um grupo de jovens rappers negros que fundou uma associação no final dos anos 1990 em Campinas, SP. Por meio de longas, repetidas entrevistas e observações de alguns dos seus encontros e práticas, mostramos que a associação foi o resultado de um processo de aprendizagem por que passaram os jovens, ao longo de um contato estreito com políticos profissionais mais velhos e mais experientes. Nesse processo eles aprenderam como um movimento de protesto deveria se articular para influenciar o governo. Aprenderam também certas técnicas de mobilização que lhes permitiram ser respeitados e construir apoios em outros segmentos de ativismo partidário e sindical. As ações da associação deram concretude e visibilidade à idéia de que havia um "movimento hip hop" na cidade. Em 2001, ao final de uma longa negociação, seus membros obtiveram o apoio da prefeitura municipal da cidade, assumida pelo PT, para a criação da Casa do Hip Hop de Campinas, onde eles foram empregados para desenvolver atividades artísticas e políticas.
A vocação pedagógica dos museus de Filadélfia
Authors: Magalhães Roberto Carvalho de
Published: September 2009

A ação educativa dos museus de Filadélfia é extraordinariamente ampla. Dos museus menores, como o Benjamin Franklin Underground Museum, ao imenso Philadelphia Museum of Art, eles oferecem um leque vasto e diversificado de materiais pedagógicos, seja dirigidos à escola, seja ao público em geral. Este ensaio relata e analisa as suas estratégias educativas, especialmente as ações educativas dos museus de arte.
Descrição de dois anos de atuação da Comissão de Ética no Uso de Animais do Setor de Ciências Agrárias da Universidade Federal do Paraná
Authors: Silla Vanessa Carli Bones , Marthos Silmara Maldonado , Molento Carla Forte Maiolino
Published: September 2009

A Comissão de Ética no Uso de Animais do Setor de Ciências Agrárias (CEUA-SCA) da Universidade Federal do Paraná (UFPR) foi criada em 2005. Este artigo teve por objetivo estudar os dois primeiros anos do trabalho dessa Comissão, principalmente quanto à redução do sofrimento animal. Foram avaliadas a composição e a metodologia de trabalho da CEUA-SCA, além da efetividade na redução do sofrimento animal, pela razão emissão de certificados por projetos protocolados (C:P) e pelo estudo dos projetos que receberam ofícios. Foram avaliados 42 protocolos de pesquisa de 2006 e 24 de 2007. Animais vertebrados foram os mais usados, submetidos com maior frequência a procedimentos grau B de invasividade. Os resultados sugerem que a CEUA-SCA, no período estudado, colaborou para o controle do uso de animais nas pesquisas sob a responsabilidade do SCA-UFPR. Destaca-se a necessidade do preenchimento adequado dos formulários pelos proponentes, o aumento da pluralidade na composição da Comissão e a importância de um órgão federal que centralize informações relativas à atuação das CEUAs no Brasil.
Política e economia na ação coletiva: uma crítica etnográfica às premissas dicotômicas
Authors: Quirós Julieta
Published: September 2009

Este artigo procura, distanciando-se dos modelos formais que têm servido de eixo para os estudos sobre ação coletiva e movimentos sociais, restituir a dimensão vivida do engajamento político. Com base em análise etnográfica centrada num conjunto de bairros da Grande Buenos Aires, explora os modos - múltiplos e heterogêneos - como as pessoas se envolvem nos chamados movimentos piqueteros, e indica que essas experiências ganham inteligibilidade ao serem inscritas numa trama mais ampla de relações e possibilidades de vida. A partir de uma perspectiva figuracional, o texto discute alguns pressupostos da literatura sobre organizações piqueteras, em particular, e protestos sociais, em geral; questiona a dicotomia entre razão material e razão político-moral com que se tem abordado a questão das motivações da ação coletiva; desconfia de uma oposição rígida entre Estado e movimentos sociais, apontando para o papel criador - e não só cooptador - das políticas estatais; e, finalmente, propõe sociologizar o lugar do "prazer de fazer" na origem e na continuidade do engajamento político.
Produção e composição química do leite, consumo e digestibilidade de forragens tropicais manejadas em sistema de lotação intermitente
Authors: Porto Petrônio Pinheiro , Deresz Fermino , Santos Geraldo Tadeu dos , Lopes Fernando César Ferraz , Cecato Ulysses , Cóser Antônio Carlos
Published: September 2009

Os objetivos neste trabalho foram comparar a composição química, a digestibilidade in vitro da matéria seca (DIVMS), as frações lâminas foliares e bainhas-colmos e a disponibilidade de forragem e avaliar a produção e composição do leite, o peso vivo, o escore corporal e o consumo de matéria seca de forrageiras tropicais. As forrageiras avaliadas foram capim-tanzânia (Panicum maximum Jacq.), grama-estrela (Cynodon nlemfuensis Vanderyst) e capim-marandu (Brachiaria brizantha Staf). As pastagens foram adubadas com nitrogênio (200 kg/ha.ano), K2O e P2O5 (50 kg/ha.ano) e manejadas em sistema de lotação intermitente com três dias de ocupação de piquete e dois intervalos de desfolha: janeiro e fevereiro com 24 dias e março, abril e maio com 30 dias. Foram utilizadas 24 vacas em delineamento de blocos ao acaso, com oito vacas por tratamento. As lâminas foliares da grama-estrela tinham em média 18,5% de PB e as bainhas-colmos, 79,9% de fibra em detergente neutro (FDN), valores superiores aos obtidos nas demais forrageiras. A grama-estela também apresentou maior teor de FDN (72,8%) e lignina (7,1%) na forragem disponível. A digestibilidade in vitro da matéria seca do capim-marandu (64,%) foi superior à da grama-estrela (59,4%). Nos capins-tanzânia e marandu, as produções de leite foram de 9,8, valor superior ao de 8,4 kg/vaca.dia da grama-estrela. O teor de proteína do leite das vacas manejadas no capim-tanzânia (2,9%) foi superior ao obtido com grama-estrela (2,8%), mas os teores de gordura e sólidos totais do leite, o escore corporal, o peso vivo e o consumo de matéria seca não diferiram entre as forrageiras avaliadas. A grama-estrela teve maior porcentagem das frações fibrosas, menor DIVMS e menor produção de leite em comparação aos capins-tanzânia e marandu.
Avaliação de indicadores em estudos com ruminantes: estimativa de consumos de concentrado e de silagem de milho por vacas em lactação
Authors: Ferreira Marcelo de Andrade , Valadares Filho Sebastião de Campos , Silva Luiz Fernando Costa e , Nascimento Fernando Barros , Detmann Edenio , Valadares Rilene Ferreira Diniz
Published: September 2009

Dois trabalhos foram conduzidos para estimativa do consumo individual de vacas em lactação alimentadas em grupo. No primeiro, para validar a metodologia, foram utilizadas oito vacas, alojadas em baias individuais recebendo silagem de milho e 4 kg de concentrado. Dois indicadores externos (óxido crômico e dióxido de titânio) foram avaliados para estimativa do consumo individual de concentrado. A fibra em detergente ácido indigestível (FDAi) foi utilizada para estimativa do consumo de silagem. A produção de matéria seca fecal (PMSF) foi determinada pela coleta total de fezes e estimada pelo LIPE®. Tanto o óxido crômico quanto o dióxido de titânio permitiram estimar com eficiência o consumo de concentrado, independentemente do método para estimativa da PMSF. A FDAi estimou de forma satisfatória o consumo de silagem de milho. No segundo experimento, foram utilizadas 31 vacas, distribuídas em três grupos de alimentação, alojadas em estábulo tipo free stall e alimentadas com silagem de milho e concentrado de acordo com a produção de leite (8,0; 5,5 e 4 kg, para os grupos 1, 2 e 3, respectivamente). Não houve diferença entre o óxido crômico e o dióxido de titânio no consumo de MS do concentrado, que foi muito próximo da quantidade média fornecida por vaca/dia (6,99 vs 7,12; 4,81 vs 4,96 e 3,49 vs 3,52 kg/vaca/dia). Verificou-se grande variação individual no consumo de concentrado, silagem e matéria seca total, independentemente do grupo de alimentação. Não foi verificada relação entre o consumo de matéria seca, a produção de leite e o peso metabólico dos animais. O óxido crômico e o dióxido de titânio podem ser usados para estimativa do consumo individual de concentrado e a FDAi é adequada para estimativa do consumo de silagem de milho por vacas em lactação alimentadas em grupo.
Importance of retardation and fatigue/interest domains for the diagnosis of major depressive episode after stroke: a four months prospective study
Authors: Terroni Luisa de Marillac Niro , Fráguas Renério , Lucia Mara de , Tinone Gisela , Mattos Patricia , Iosifescu Dan V. , Scaf Milberto
Published: September 2009

OBJECTIVE: Post-stroke major depressive episode is very frequent, but underdiagnosed. Researchers have investigated major depressive episode symptomatology, which may increase its detection. This study was developed to identify the depressive symptoms that better differentiate post-stroke patients with major depressive episode from those without major depressive episode. METHOD: We screened 260 consecutive ischemic stroke patients admitted to the neurology clinic of a university hospital. Seventy-three patients were eligible and prospectively evaluated. We assessed the diagnosis of major depressive episode using the Structured Clinical Interview for DSM-IV and the profile of depressive symptoms using the 31-item version of the Hamilton Depression Rating Scale. For data analysis we used cluster analyses and logistic regression equations. RESULTS: Twenty-one (28.8%) patients had a major depressive episode. The odds ratio of being diagnosed with major depressive episode was 3.86; (95% CI, 1.23-12.04) for an increase of one unit in the cluster composed by the domains of fatigue/interest and retardation, and 2.39 (95% CI, 1.21-4.71) for an increase of one unit in the cluster composed by the domains of cognitive, accessory and anxiety symptoms. The domains of eating/weight and insomnia did not contribute for the major depressive episode diagnosis. CONCLUSION: The domains of retardation and interest/fatigue are the most relevant for the diagnosis of major depressive episode after stroke.
Avaliação nutricional e autopercepção corporal de praticantes de musculação em academias de Caxias do Sul - RS
Authors: Theodoro Heloísa , Ricalde Simone Rufatto , Amaro Francisco Stefani
Published: September 2009

O objetivo deste estudo foi diagnosticar o estado nutricional e a autopercepção corporal de frequentadores de academias de Caxias do Sul. A amostra foi composta por 87 homens, na faixa etária entre 20 e 25 anos. Para o diagnóstico do estado nutricional foi feita a análise da adequação da ingestão de macronutrientes e micronutrientes - vitaminas A, C e E, cálcio, ferro, selênio e zinco, por meio do método recordatório de 24 horas, além da caracterização do perfil antropométrico e da autopercepção corporal, que foi avaliada através de um questionário com questões fechadas e escalas de silhuetas. Para análise estatística foram utilizadas análise descritiva e o teste t de Student pareado. Valores de p < 0,05 foram considerados significantes. Os resultados indicaram que a alimentação dessa população foi caracterizada como normoglicídica, hiperlipídica e hiperproteica; o consumo de suplementos foi de 29,9%, sendo a maioria à base de proteínas. Quanto aos micronutrientes, mostrou adequação em selênio, zinco e ferro e inadequação em vitaminas A, C e E, e cálcio. Apenas 19,5% dos indivíduos ingeriram o recomendado desse mineral por dia. O percentual de gordura corporal dos praticantes de musculação (14,08 ± 4,25%) foi classificado como dentro da média. Em relação à autopercepção corporal, a maioria deseja ter uma silhueta igual a ou maior que a representada por um indivíduo eutrófico. Essa população apresenta importantes inadequações nutricionais, sendo ou não decorrente do objetivo de ganho de massa muscular, necessitando assim de orientação nutricional.
Extreme storm surges in the south of Brazil: atmospheric conditions and shore erosion
Authors: Parise Cláudia Klose , Calliari Lauro Júlio , Krusche Nisia
Published: September 2009

The region under study is regularly subject to the occurrence of storms associated with frontal systems and extratropical cyclones, since it is located near one of the cyclogenetic regions in South America. These storms can generate storm surges that cause anomalous high sea level rises on Cassino Beach. The use of reanalysis data along with an efficient technique for the location of the cyclone, using a vorticity threshold, has provided a new classification based upon the trajectories of events that produce positive sea level variation. Three patterns have been identified: 1) Cyclogenesis to the south of Argentina with displacement to the east and a trajectory between 47.5ºS and 57.5ºS; 2) Cyclogenesis to the south of Uruguay with displacement to the east and a trajectory between 35ºS and 42.5ºS; and 3) Cyclogenesis to the south of Uruguay with displacement to the southeast and a trajectory between 35ºS and 57.5ºS. Maximum water level elevation above the mean sea level and beach erosion were associated, respectively, with winter and summer storms. Cassino beach displayed a seasonal morphological behavior, with short periods of episodic erosion associated with winter storm events followed by long periods of accretion characterized by the dominance of fair weather conditions.
The Effects of Adolescent Health on Educational Outcomes: Causal Evidence Using Genetic Lotteries between Siblings
Authors: Fletcher Jason M. , Lehrer Steven F
Published: September 2009

There has been growing interest in using specific genetic markers as instrumental variables in attempts to assess causal relationships between health status and socioeconomic outcomes, including human capital accumulation. In this paper, we use a combination of family fixed effects and genetic marker instruments to estimate the causal effects of poor adolescent mental and physical health status on years of completed schooling. By exploiting the genetic variation in inheritance within families, this empirical strategy presents a unique opportunity to isolate the variation in genetic factors from other dynastic and family characteristics. We present evidence that inattentive symptoms in early childhood have large lasting effects in reducing completed schooling. We also find little consistent evidence that adolescent overweight status influences years of schooling completed.
Climate Change and Conservation in Brazil: CGE Evaluation of Health and Wealth Impacts
Authors: Pattanayak Subhrendu K. , Ross Martin T. , Depro Brooks M. , Bauch Simone C. , Timmins Christopher , Wendland Kelly J. , Alger Keith
Published: September 2009

Ecosystem services are public goods that frequently constitute the only source of capital for the poor, who lack political voice. As a result, provision of ecosystem services is sub-optimal and estimation of their values is complicated. We examine how econometric estimation can feed computable general equilibrium (CGE) modeling to estimate health-related ecosystem values. Against a back drop of climate change, we analyze the Brazilian policy to expand National Forests (FLONAS) by 50 million hectares. Because these major environmental changes can generate spillovers in other sectors, we develop and use a CGE model that focuses on land and labor markets. Compared to climate change and deforestation in the baseline, the FLONAS scenario suggests relatively small declines in GDP, output (including agriculture) and other macro indicators. Urban households will experience declines in their welfare because they own most of the capital and land, which allows them to capture most of the deforestation benefits. In contrast, even though rural households have fewer opportunities for subsistence agriculture and face additional competition with other rural agricultural workers for more limited employment, their welfare improves due to health benefits from conservation of nearby forests. The efficiency vs. equity tradeoffs implied by the FLONAS scenario suggests that health-related ecosystem services will be underprovided if the rural poor are politically weaker than the urban rich. In conclusion, we briefly discuss the pros and cons of the CGE strategy for valuing ecosystem-mediated health benefits and evaluating contemporary policies on climate change mitigation.
Analysis of Revival Phenomenon for Strong-Field Excitation of Rydberg Atom
Authors: Veilande R. , Bersons I.
Published: September 2009

Analysis of Revival Phenomenon for Strong-Field Excitation of Rydberg Atom

The autocorrelation function for a one-dimensional Rydberg atom irradiated by half-cycle pulse (HCP) is investigated. At weak field only the full revival is observed, at a medium field strength - a fractional revival, whereas at a strong field (when the continuous energy states are also populated) the revival phenomenon disappears. The contribution of the continuous energy states in the autocorrelation function decreases dramatically with time after the wave packet formation.
Stopover Ecology of Some Passerines at Ankara (Central Turkey)
Authors: Keşaplı Can Özge , Can Bilgin Cemal
Published: September 2009

Stopover Ecology of Some Passerines at Ankara (Central Turkey)

In autumn 2002 we studied little known passerine migration at a woodland patch within the Middle East Technical University (METU) campus in Ankara (Turkey). A total of 954 individuals of 35 passerine species were mist-netted, ringed, measured, weighed and fat-scored (after Busse 2000). Blackcap (Sylvia atricapilla) and Willow Warbler (Phylloscopus trochilus) were the two most common species, with 308 and 145 individuals caught, respectively. Both are passage migrants at METU, recorded from mid-August to late October, representing several waves.

Only 11.5% of Blackcaps had the fat score of T6
and above, and among 20 retraps only 2 gained fat significantly. All the other retraps lost fat, stayed the same, or increased 1-2 scores at most. In contrast, Willow Warblers, as true trans-Saharan migrants, had the much higher proportion (46%) of individuals with fat scores of T6
and above. Most retrapped individuals gained fat, some with already high levels stayed the same, while none lost fat. We interpret these data in terms of known migratory ranges, diet types and habitat patch quality.

Although daily catches were low, a diverse range of species used METU as a stopover site. Fat deposition rates (of up to 50% of body weight within a week) suggest that the study site provided a high quality stopover habitat for most migrants. In Central Turkey, such suitable habitats with trees or tall shrubs are scarce, and therefore, crucial for migrants.
Revolution and the End of History: Caryl Churchill'sMad Forest
Authors: Adiseshiah Siân
Published: September 2009

Caryl Churchill's Mad Forest, written and performed very soon after the Romanian revolution in 1990 and performed both in London and Bucharest, is a dynamic, inter-cultural play that represents a variety of perspectives on the revolutionary events, as well as oscillating between English and Romanian cultural and language coordinates. It has a peculiar topicality in its detailed and specific uses of different aspects of the revolutionary narrative, its sketches of family life before and after the revolution, and its inclusion of the revolution as reported in quasi-documentary-style testimony. The perspective in this article is one that places the play within a framework that thinks through Mad Forest's relationship to the triumphant, neo-liberalist heralding of “the end of history,” most famously argued by Francis Fukuyama in his 1989 article of that name. This discourse gained further confidence from the collapse of Eastern Europe, a collapse that was viewed by proponents of the end-of-history argument as signalling the permanent disintegration of communism and a victory for capitalism. However, Mad Forest is considered here as a play that reflects multiple perspectives on the revolutionary period and, while declining to provide political solutions as such, simultaneously refuses to accede to the implications of the end-of-history argument.
Making the “Unstageable” Stageable: English Rewritings of Chekhov's First Play
Authors: Young Stuart
Published: September 2009

This essay examines three adaptations for the English stage of Chekhov's “unstageable” first play. As well as reflecting something of the broader context of the translation and reception of foreign plays in the English theatre, these adaptations are very illuminating on fundamental aspects of English Chekhov tradition, in particular its largely veiled politics. Michael Frayn's Wild Honey, which highlights connexions with Chekhov's later plays and reinscribes cardinal elements of English Chekhov, exemplifies the strategy of domesticating the foreign. In sharp contrast, taking issue with a tradition that sentimentalizes and de-politicizes Chekhov, Trevor Griffiths's Piano reconceives Untitled Play as a political play that comments on contemporary Britain. Meanwhile, David Hare's Platonov, which purports to stick as closely as possible to the original text, ends up, like Wild Honey, reiterating the familiar hallmarks of English Chekhov. The choices made in all three adaptations attest to the fundamentally political nature of the translator's intervention as well as to the translator's contribution not merely to the transmission of culture but to its production.
Before Katrina: Archiving Performative Downpours and Fallen Women Named Sadie inRainandThe Deluge(1922)
Authors: Johnson Katie N.
Published: September 2009

In 1922, two popular plays – Rain and The Deluge – literally took the stage by storm. In what may strike post-Katrina audiences as a somewhat uncanny coincidence, these plays were based on the aftermaths of natural disasters: specifically, torrential downpours and the breaching of a levee. Both plays also featured a down-on-her-luck “fallen” woman called Sadie. Indeed, after this season, Sadie became an iconic character-type, appearing in various Broadway hits, musicals, and films for years. In focusing on just two of the performative iterations of Sadie, this essay shows that, at the same time that Sadie rivals heteronormative sexuality (as well as codified notions of racialization), so too she finds herself in narratives that often reinscribe such ideologies. And yet, while, narratively, efforts are made to contain Sadie, such efforts often fail. Performatively, then, Sadie is a figure that often resists these oppressions and performs sexual subversions, if only momentary ones.
Extrapulmonary Tuberculosis: Experience in Rajshahi Chest Disease Clinic and Chest Disease Hospital
Authors: Hussain Md Wasim , Haque Md Azizul , Banu Salma Arjumand , Ekram ARM Saifuddin , Rahman M Fazlur
Published: September 2009

The study was carried out to see the prevalence and pattern of extrapulmonary tuberculosis in Rajshahi. Twenty five hundred eighty two tuberculosis patients (sputum positive, sputum negative and extrapulmonary) who were diagnosed and treated in between January 1999 and December 2003 in Rajshahi Chest Disease Clinic and Chest Disease Hospital were included in this study. Extrapulmonary tuberculosis accounted for about 16% of total tuberculosis cases. Tuberculous lymphadenitis was found to be the commonest form of extrapulmonary tuberculosis, comprising about 75% of the extrapulmonary cases. Tuberculous pleural effusion was noted in about 9% of cases, intestinal tuberculosis in 7%, bone and joint tuberculosis in 5% and other forms in 4% cases. 24% patients with extrapulmonary tuberculosis were afebrile, 14% had normal ESR, and 22% had negative Mantoux test.   doi: 10.3329/taj.v17i1.3484 TAJ 2004; 17(1) : 16-19
Primary Segmental Colonic Tuberculosis: A Case Report
Authors: Haque MM , Siddique AB , Badruddoza SM , Amin MR , Rahman MG , Hasan MZ , Abedin MJ , Islam M Takbirul
Published: September 2009

Tuberculosis is known to involve any segment of gastrointestinal tract. While tuberculosis of the small bowel is common, isolated primary tuberculosis of the large bowel is quite uncommon. We report two cases of isolated colonic tuberculosis, successfully treated with primary resection.   doi: 10.3329/taj.v17i1.3491 TAJ 2004; 17(1) : 52-54
Anti-Siglec-F Antibody Reduces Allergen-Induced Eosinophilic Inflammation and Airway Remodeling
Authors: Song Dae Jin , Cho Jae Youn , Lee Sang Yeub , Miller Marina , Rosenthal Peter , Soroosh Pejman , Croft Michael , Zhang Mai , Varki Ajit , Broide David H.
Published: September 2009

Siglec-F is a sialic acid-binding Ig superfamily receptor that is highly expressed on eosinophils. We have investigated whether administration of an anti-Siglec-F Ab to OVA-challenged wild-type mice would reduce levels of eosinophilic inflammation and levels of airway remodeling. Mice sensitized to OVA and challenged repetitively with OVA for 1 mo who were administered an anti-Siglec-F Ab had significantly reduced levels of peribronchial eosinophilic inflammation and significantly reduced levels of subepithelial fibrosis as assessed by either trichrome staining or lung collagen levels. The anti-Siglec-F Ab reduced the number of bone marrow, blood, and tissue eosinophils, suggesting that the anti-Siglec-F Ab was reducing the production of eosinophils. Administration of a F(ab′)2 fragment of an anti-Siglec-F Ab also significantly reduced levels of eosinophilic inflammation in the lung and blood. FACS analysis demonstrated increased numbers of apoptotic cells (annexin V+/CCR3+ bronchoalveolar lavage and bone marrow cells) in anti-Siglec-F Ab-treated mice challenged with OVA. The anti-Siglec-F Ab significantly reduced the number of peribronchial major basic protein+/TGF-β+ cells, suggesting that reduced levels of eosinophil-derived TGF-β in anti-Siglec-F Ab-treated mice contributed to reduced levels of peribronchial fibrosis. Administration of the anti-Siglec-F Ab modestly reduced levels of periodic acid-Schiff-positive mucus cells and the thickness of the smooth muscle layer. Overall, these studies suggest that administration of an anti-Siglec-F Ab can significantly reduce levels of allergen-induced eosinophilic airway inflammation and features of airway remodeling, in particular subepithelial fibrosis, by reducing the production of eosinophils and increasing the number of apoptotic eosinophils in lung and bone marrow.
Prevention of Inflammatory Activation of Human Gestational Membranes in an Ex Vivo Model Using a Pharmacological NF-κB Inhibitor
Authors: Keelan Jeffrey A. , Khan Sadiqa , Yosaatmadja Francisca , Mitchell Murray D.
Published: September 2009

Intrauterine inflammation plays a major role in the etiology of preterm labor and birth. We established an ex vivo model employing perfused full-thickness term gestational membranes to study membrane transport, function, and inflammatory responses. Exposure of the maternal (decidual) face of the membranes to LPS (5 μg/ml) resulted in increased accumulation of proinflammatory cytokines in the maternal compartment within 4 h, followed by a response in the fetal (amniotic) compartment. Using cytokine arrays, exposure to LPS was found to result in increased secretion of a large number of cytokines and chemokines in both compartments, most notably IL-5, IL-6, IL-7, MDC (macrophage-derived chemokine), MIG (monokine induced by IFN-γ), TARC (thymus and activation-regulated chemokine), TGF-β, and TNF-α. PGE2 accumulation also increased in response to LPS, particularly in the fetal compartment. Cotreatment with sulfasalazine, which inhibited nuclear translocation of NF-κB p65, had a rapid and marked inhibitory effect on the rate of cytokine accumulation in the maternal compartment, with lesser but significant effects observed in the fetal compartment. While membrane integrity was not discernibly impaired with LPS or sulfasalazine exposure, rates of chorionic apoptosis after 20 h were doubled in sulfasalazine-treated tissues. We conclude that the system described provides a means of accurately modeling human gestational membrane functions and inflammatory activation ex vivo. Decidual LPS exposure was shown to elicit a robust inflammatory response in both the maternal and fetal compartments. Sulfasalazine was an effective antiinflammatory agent in this model, but also exerted proapoptotic effects that raise concerns regarding its placental effects when administered in pregnancy.
MAIL Regulates Human Monocyte IL-6 Production
Authors: Seshadri Sudarshan , Kannan Yashaswini , Mitra Srabani , Parker-Barnes Jennifer , Wewers Mark D.
Published: September 2009

IL-6 is a pleiotropic cytokine implicated in the pathogenesis of disorders such as sepsis and cancer. We noted that human monocytes are excellent producers of IL-6 as compared with monocyte-derived macrophages. Because macrophages from molecule containing ankyrin repeats induced by LPS (MAIL) knockout animals have suppressed IL-6 production, we hypothesized that regulation of MAIL is key to IL-6 production in humans and may explain the differences between human monocytes and macrophages. To test this hypothesis fresh human monocytes and monocyte-derived macrophages were compared for MAIL expression in response to LPS. LPS-induced monocyte MAIL expression was highly inducible and transient. Importantly for our hypothesis MAIL protein expression was suppressed during differentiation of monocytes to macrophages. Of note, the human MAIL protein detected was the 80 kDa MAIL-L form and human MAIL showed nuclear localization. Human MAIL-L bound to p50 subunit of the NF-κB and increased IL-6 luciferase promoter activity in a cEBPβ, NF-κB, and AP-1-dependent fashion. Like the differences in MAIL induction, monocytes produced 6-fold more IL-6 compared with macrophages (81.7 ± 29.7 vs 12.6 ± 6.8 ng/ml). Furthermore, suppression of MAIL by small interfering RNA decreased the production of IL-6 significantly in both THP-1 cells and in primary monocytes. Costimulation of monocytes with LPS and muramyl dipeptide induced an enhanced IL-6 response that was suppressed by siMAIL. Our data suggests that MAIL is a key regulator of IL-6 production in human monocytes and plays an important role in both TLR and NOD-like receptor ligand induced inflammation.
Lymphoid Enhancer Binding Factor 1 Regulates Transcription through Gene Looping
Authors: Yun Kangsun , So Jae-Seon , Jash Arijita , Im Sin-Hyeog
Published: September 2009

Efficient transcription depends upon efficient physical and functional interactions between transcriptosome complexes and DNA. We have previously shown that IL-1β-induced lymphoid enhancer binding factor 1 (Lef1) regulates the transcription of its target genes COX2 and MMP13 in mouse chondrocytes by binding to the Lef1 binding sites located in the 3′ region. In this study, we investigated how the 3′ region-bound Lef1 regulates expression of target genes. IL-1β stimulation induced gene looping in COX2 and MMP13 genomic loci, which is mediated by the physical interaction of Lef1 with its binding partners, including β-catenin, AP-1, and NF-κB. As shown by chromosome conformation capture (3C) assay, the 5′ and 3′ genomic regions of these genes were juxtaposed in an IL-1β-stimulation dependent manner. Lef1 played a pivotal role in this gene looping; Lef1 knockdown decreased the incidence of gene looping, while Lef1 overexpression induced it. Physical interactions between the 3′ region-bound Lef1 and promoter-bound transcription factors AP-1 or NF-κB in COX2 and MMP13, respectively, were increased upon stimulation, leading to synergistic up-regulation of gene expression. Knockdown of RelA or c-Jun decreased the formation of gene loop and down-regulated cyclooxygenase 2 (COX2) or matrix metalloproteinase 13 (MMP13) transcription levels. However, overexpression of RelA or c-Jun along with Lef1 increased the looping and their expression levels. Our results indicate a novel function of Lef1, as a mediator of gene looping between 5′ and 3′ regions. Gene looping may serve to delineate the transcription unit in the inducible gene transcription of mammalian cells.
Development of Allergen-Induced Airway Inflammation in the Absence of T-bet Regulation Is Dependent on IL-17
Authors: Durrant Douglas M. , Gaffen Sarah L. , Riesenfeld Erik P. , Irvin Charles G. , Metzger Dennis W.
Published: September 2009

Dysfunctional expression of T-bet, a transcription factor that is critical for IFN-γ production, has been implicated in the development of asthma. To investigate in detail the mechanisms responsible for exacerbated disease in the absence of T-bet expression, BALB/c wild-type (WT) and T-bet−/− mice were used in a murine model of OVA-induced allergic lung inflammation. Following OVA challenge, T-bet−/− mice displayed increased histological inflammation in the lungs as well as greater thickening of the bronchiole linings, increased numbers of eosinophils and neutrophils in the lung, and enhanced airway hyperresponsiveness, compared with WT mice. However, the production of Th2 cytokines in T-bet−/− mice did not appear to be significantly greater than in WT mice. Interestingly, a marked increase in the levels of the proinflammatory cytokine IL-17 was observed in T-bet−/− mice. Neutralization of pulmonary IL-17 in T-bet−/− mice by anti-IL-17 mAb treatment during OVA challenge resulted in decreased levels of neutrophilic infiltration into the airways and decreased airway inflammation, essentially reversing the development of allergic asthma development. These findings indicate that IL-17 is a key mediator of airway inflammation in the absence of T-bet. The results of this study suggest a possible target for therapeutic intervention of human asthma.
IFN-β Inhibits Human Th17 Cell Differentiation
Authors: Ramgolam Vinod S. , Sha Yonggang , Jin Jianping , Zhang Xin , Markovic-Plese Silva
Published: September 2009

IFN-β-1a has been used over the past 15 years as a primary therapy for relapsing-remitting multiple sclerosis (MS). However, the immunomodulatory mechanisms that provide a therapeutic effect against this CNS inflammatory disease are not yet completely elucidated. The effect of IFN-β-1a on Th17 cells, which play a critical role in the development of the autoimmune response, has not been extensively studied in humans. We have investigated the effect of IFN-β-1a on dendritic cells (DCs) and naive CD4+CD45RA+ T cells derived from untreated MS patients and healthy controls in the context of Th17 cell differentiation. We report that IFN-β-1a treatment down-regulated the expression of IL-1β and IL-23p19 in DCs, whereas it induced the gene expression of IL-12p35 and IL-27p28. We propose that IFN-β-1a-mediated up-regulation of the suppressor of cytokine signaling 3 expression, induced via STAT3 phosphorylation, mediates IL-1β and IL-23 down-regulation, while IFN-β-1a-induced STAT1 phosphorylation induces IL-27p28 expression. CD4+CD45RA+ naive T cells cocultured with supernatants from IFN-β-1a-treated DCs exhibited decreased gene expression of the Th17 cell markers retinoic acid-related orphan nuclear hormone receptor c (RORc), IL-17A, and IL-23R. A direct IFN-β-1a treatment of CD45RA+ T cells cultured in Th17-polarizing conditions also down-regulated RORc, IL-17A, and IL-23R, but up-regulated IL-10 gene expression. Studies of the mechanisms involved in the Th17 cell differentiation suggest that IFN-β-1a inhibits IL-17 and induces IL-10 secretion via activated STAT1 and STAT3, respectively. IFN-β’s suppression of Th17 cell differentiation may represent its most relevant mechanism of selective suppression of the autoimmune response in MS.
IL-4Rα, a New Member that Associates with Syk Kinase: Implication in IL-4-Induced Human Neutrophil Functions
Authors: Ennaciri Jamila , Girard Denis
Published: September 2009

Although Syk has been reported to be associated with IL-2Rα and IL-15Rα in some hematopoietic cells, its association has never been investigated in the IL-4/IL-4R system. In this study, we demonstrate for the first time that Syk is constitutively associated with IL-4Rα in human polymorphonuclear neutrophils (PMNs) and that IL-4 stimulation increases the amount of Syk associated with IL-4Rα. Moreover, upon IL-4 treatment, a pool of Syk associated with IL-4Rα is phosphorylated. We also report that such association is not unique to PMNs because Syk associates with IL-4Rα in Raji and in PBMC cells. Stimulation of PMNs by IL-4 increased the amount of Syk associated with PLC-γ2, pAkt, and α-tubulin. Pretreatment of cells with the Syk-selective inhibitor piceatannol or Syk inhibitor II, significantly inhibited the ability of IL-4 to enhance phagocytosis and cell adhesion and to delay apoptosis, and these results correlate with the ability of piceatannol to reduce Syk activation and its association with IL-4Rα. Down-regulation of Syk by antisense techniques demonstrates the importance of Syk in the antiapoptotic effect of IL-4. We conclude that association of Syk to IL-4Rα is of biological significance and that IL-4Rα is a new candidate to be added to the few cytokine receptor components which associate with Syk.
Identification of Mycobacterial α-Glucan As a Novel Ligand for DC-SIGN: Involvement of Mycobacterial Capsular Polysaccharides in Host Immune Modulation
Authors: Geurtsen Jeroen , Chedammi Sunita , Mesters Joram , Cot Marlène , Driessen Nicole N. , Sambou Tounkang , Kakutani Ryo , Ummels Roy , Maaskant Janneke , Takata Hiroki , Baba Otto , Terashima Tatsuo , Bovin Nicolai , Vandenbroucke-Grauls Christina M. J. E. , Nigou Jérôme , Puzo Germain , Lemassu Anne , Daffé Mamadou , Appelmelk Ben J.
Published: September 2009

Mycobacterium tuberculosis possesses a variety of immunomodulatory factors that influence the host immune response. When the bacillus encounters its target cell, the outermost components of its cell envelope are the first to interact. Mycobacteria, including M. tuberculosis, are surrounded by a loosely attached capsule that is mainly composed of proteins and polysaccharides. Although the chemical composition of the capsule is relatively well studied, its biological function is only poorly understood. The aim of this study was to further assess the functional role of the mycobacterial capsule by identifying host receptors that recognize its constituents. We focused on α-glucan, which is the dominant capsular polysaccharide. Here we demonstrate that M. tuberculosis α-glucan is a novel ligand for the C-type lectin DC-SIGN (dendritic cell-specific ICAM-3-grabbing nonintegrin). By using related glycogen structures, we show that recognition of α-glucans by DC-SIGN is a general feature and that the interaction is mediated by internal glucosyl residues. As for mannose-capped lipoarabinomannan, an abundant mycobacterial cell wall-associated glycolipid, binding of α-glucan to DC-SIGN stimulated the production of immunosuppressive IL-10 by LPS-activated monocyte-derived dendritic cells. By using specific inhibitors, we show that this IL-10 induction was DC-SIGN-dependent and also required acetylation of NF-κB. Finally, we demonstrate that purified M. tuberculosis α-glucan, in contrast to what has been reported for fungal α-glucan, was unable to activate TLR2.
Critical Role of IL-17RA in Immunopathology of Influenza Infection
Authors: Crowe Christopher R. , Chen Kong , Pociask Derek A. , Alcorn John F. , Krivich Cameron , Enelow Richard I. , Ross Ted M. , Witztum Joseph L. , Kolls Jay K.
Published: September 2009

Acute lung injury due to influenza infection is associated with high mortality, an increase in neutrophils in the airspace, and increases in tissue myeloperoxidase (MPO). Because IL-17A and IL-17F, ligands for IL-17 receptor antagonist (IL-17RA), have been shown to mediate neutrophil migration into the lung in response to LPS or Gram-negative bacterial pneumonia, we hypothesized that IL-17RA signaling was critical for acute lung injury in response to pulmonary influenza infection. IL-17RA was critical for weight loss and both neutrophil migration and increases in tissue myeloperoxidase (MPO) after influenza infection. However, IL-17RA was dispensable for the recruitment of CD8+ T cells specific for influenza hemagglutinin or nucleocapsid protein. Consistent with this, IL-17RA was not required for viral clearance. However, in the setting of influenza infection, IL-17RA−/− mice showed significantly reduced levels of oxidized phospholipids, which have previously been shown to be an important mediator in several models of acute lung injury, including influenza infection and gastric acid aspiration. Taken together, these data support targeting IL-17 or IL-17RA in acute lung injury due to acute viral infection.
CCR7 Deficiency on Dendritic Cells Enhances Fungal Clearance in a Murine Model of Pulmonary Invasive Aspergillosis
Authors: Hartigan Adam J. , Westwick John , Jarai Gabor , Hogaboam Cory M.
Published: September 2009

Aspergillus fumigatus is a sporulating fungus found ubiquitously in the environment and is easily cleared from immunocompetent hosts. Invasive aspergillosis develops in immunocompromised patients, and is a leading cause of mortality in hematopoietic stem cell transplant recipients. CCR7 and its ligands, CCL19 and CCL21, are responsible for the migration of dendritic cells from sites of infection and inflammation to secondary lymphoid organs. To investigate the role of CCR7 during invasive aspergillosis, we used a well-characterized neutropenic murine model. During invasive aspergillosis, mice with a CCR7 deficiency in the hematopoietic compartment exhibited increased survival and less pulmonary injury compared with the appropriate wild-type control. Flow cytometric analysis of the chimeric mice revealed an increase in the number of dendritic cells present in the lungs of CCR7-deficient chimeras following infection with Aspergillus conidia. An adoptive transfer of dendritic cells into neutropenic mice provided a protective effect during invasive aspergillosis, which was further enhanced with the adoptive transfer of CCR7-deficient dendritic cells. Additionally, CCR7-deficient dendritic cells activated in vitro with Aspergillus conidia expressed higher TNF-α, CXCL10, and CXCL2 levels, indicating a more activated cellular response to the fungus. Our results suggest that the absence of CCR7 is protective during invasive aspergillosis in neutropenic mice. Collectively, these data demonstrate a potential deleterious role for CCR7 during primary immune responses directed against A. fumigatus.
Antigen-Induced Increases in Pulmonary Mast Cell Progenitor Numbers Depend on IL-9 and CD1d-Restricted NKT Cells
Authors: Jones Tatiana G. , Hallgren Jenny , Humbles Alison , Burwell Timothy , Finkelman Fred D. , Alcaide Pilar , Austen K. Frank , Gurish Michael F.
Published: September 2009

Pulmonary mast cell progenitor (MCp) numbers increase dramatically in sensitized and aerosolized Ag-challenged mice. This increase depends on CD4+ T cells, as no MCp increase occurs in the lungs of sensitized wild-type (WT) mice after mAb depletion of CD4+ but not CD8+ cells before aerosol Ag challenge. Neither the genetic absence of IL-4, IL-4Rα chain, STAT-6, IFN-γ, or IL-12p40 nor mAb blockade of IFN-γ, IL-3, IL-4, IL-5, IL-6, IL-10, IL-13, IL-17A, IL-12p40, or IL-12p40Rβ1 before Ag challenge in WT mice reduces the pulmonary MCp increase. However, sensitized and Ag-challenged IL-9-deficient mice and sensitized WT mice given mAb to IL-9 just before Ag challenge show significant reductions in elicited lung MCp/106 mononuclear cells of 47 and 66%, respectively. CD1d-deficient mice and WT mice receiving anti-CD1d before Ag challenge also show significant reductions of 65 and 59%, respectively, in elicited lung MCp/106 mononuclear cells, revealing an additional requirement for MCp recruitment. However, in Jα18-deficient mice, which lack only type 1 or invariant NKT cells, the increase in the numbers of lung MCp with Ag challenge was intact, indicating that their recruitment must be mediated by type 2 NKT cells. Furthermore, anti-CD1d treatment of IL-9-deficient mice or anti-IL-9 treatment of CD1d-deficient mice does not further reduce the significant partial impairment of MCp recruitment occurring with a single deficiency. These findings implicate type 2 NKT cells and IL-9 as central regulators that function in the same pathway mediating the Ag-induced increase in numbers of pulmonary MCp.