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1.
Sci Data ; 8(1): 141, 2021 05 26.
Artigo em Inglês | MEDLINE | ID: mdl-34040008

RESUMO

We report high resolution measurements of the stable isotope ratios of ancient ice (δ18O, δD) from the North Greenland Eemian deep ice core (NEEM, 77.45° N, 51.06° E). The record covers the period 8-130 ky b2k (y before 2000) with a temporal resolution of ≈0.5 and 7 y at the top and the bottom of the core respectively and contains important climate events such as the 8.2 ky event, the last glacial termination and a series of glacial stadials and interstadials. At its bottom part the record contains ice from the Eemian interglacial. Isotope ratios are calibrated on the SMOW/SLAP scale and reported on the GICC05 (Greenland Ice Core Chronology 2005) and AICC2012 (Antarctic Ice Core Chronology 2012) time scales interpolated accordingly. We also provide estimates for measurement precision and accuracy for both δ18O and δD.

2.
Exp Gerontol ; 45(10): 779-87, 2010 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-20600781

RESUMO

The purpose of this study was to evaluate the relevance of long-term endothelial cell culture as a model system of vascular ageing. Micro- and macrovascular endothelial cells were serially passaged until replicative senescence and their ability to form tube-like structures when cultured on Matrigel was assessed throughout their lifespan. For both cell types low passage cultures adopted a homogeneous cobblestone morphology, while senescent cultures were extremely heterogeneous. Furthermore, both cell types showed a reduction in tube formation ability with in vitro ageing, which is in accordance with the reduction in angiogenic potential observed with ageing in vivo. Examination of senescence associated ß-galactosidase activity revealed an increased activity in cells forming tubes as compared to cells cultured on plastic, which could be attributed to an increased lysosomal content of cells undergoing tube formation. As this increased senescence associated ß-galactosidase activity was unrelated to the replicative age of the cells, senescence associated ß-galactosidase activity may not be a relevant senescence marker for differentiating endothelial cells. The age-related reduction in tube formation ability suggested that long-term culture of endothelial cells may be a valid model system of vascular ageing, which makes it an ideal platform for high throughput screening of compounds influencing angiogenesis.


Assuntos
Envelhecimento/fisiologia , Senescência Celular/fisiologia , Células Endoteliais/citologia , Microvasos/citologia , Microvasos/fisiologia , Adulto , Biomarcadores/metabolismo , Diferenciação Celular/fisiologia , Divisão Celular/fisiologia , Células Cultivadas , Dano ao DNA/fisiologia , Derme/irrigação sanguínea , Humanos , Lisossomos/metabolismo , Modelos Biológicos , beta-Galactosidase/metabolismo
3.
FEMS Immunol Med Microbiol ; 40(2): 129-37, 2004 Mar 08.
Artigo em Inglês | MEDLINE | ID: mdl-14987731

RESUMO

Chlamydia pneumoniae is an obligate intracellular bacterium that causes upper and lower respiratory tract infection in humans. C. pneumoniae harbors the polymorphic membrane protein (Pmp) family with 21 different proteins with a molecular mass around 100 kDa. The Pmps are species-specific, abundant and, together with major outer membrane protein and outer membrane protein 2, the dominant proteins in the C. pneumoniae outer membrane complex. Nevertheless, it is unknown whether Pmps are recognized by the cell-mediated immune response. To address this issue, C57BL/6J mice were infected intranasally with C. pneumoniae and the immune response to primary infection was investigated. We demonstrate, as expected, that the primary response is of the Th1 type by IgG2a- and IgG1-specific sELISA (Medac) on serum. In vivo-primed spleen lymphocytes were found to be reactive to Pmp8, Pmp20 and Pmp21 in an interferon-gamma ELISpot assay. The responses were shown to be mediated by CD4(+) T cells. To our knowledge, this is the first identification of antigens recognized by CD4(+) T cells during murine C. pneumoniae infection.


Assuntos
Antígenos de Bactérias/imunologia , Proteínas da Membrana Bacteriana Externa/imunologia , Linfócitos T CD4-Positivos/imunologia , Infecções por Chlamydophila/imunologia , Chlamydophila pneumoniae/química , Animais , Proteínas da Membrana Bacteriana Externa/química , Proteínas da Membrana Bacteriana Externa/genética , Contagem de Linfócito CD4 , Chlamydophila pneumoniae/genética , Chlamydophila pneumoniae/imunologia , Genoma Bacteriano , Camundongos , Camundongos Endogâmicos BALB C
4.
J Infect Dis ; 188(1): 108-13, 2003 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-12825178

RESUMO

We tested whether polymorphic membrane proteins (PMPs) of Chlamydia pneumoniae might play a role in triggering an inflammatory response in human endothelial cells. Of 15 purified, recombinant chlamydial PMPs tested, 2 (PMP 20 and PMP 21) dose-dependently increased the production of the inflammatory mediators interleukin (IL)-6 and monocyte chemoattractant protein-1 (MCP-1), in cultured human endothelial cells; production of IL-8 was also increased. When endothelial cells were infected by live C. pneumoniae, an increase in the production of IL-6, IL-8, and MCP-1 was seen. We used adenovirus-induced overexpression of IkappaBalpha-an inhibitor of nuclear factor (NF)-kappaB-to demonstrate that PMP 20 and PMP 21 increase the production of IL-6 and MCP-1 in human endothelial cells by activation of the NF-kappaB pathway, because, in cells overexpressing IkappaBalpha, treatment with the respective PMP did not result in increased production of IL-6 and MCP-1. Thus, C. pneumoniae could, by interactions of its PMPs with the endothelium, contribute to the process of vascular injury during the development and progression of atherosclerotic lesions.


Assuntos
Proteínas da Membrana Bacteriana Externa/farmacologia , Chlamydophila pneumoniae/química , Citocinas/biossíntese , Endotélio Vascular/efeitos dos fármacos , Endotélio Vascular/metabolismo , NF-kappa B/metabolismo , Células Cultivadas , Quimiocina CCL2/biossíntese , Humanos , Inflamação/metabolismo , Interleucina-6/biossíntese , Interleucina-8/biossíntese , Transdução de Sinais/efeitos dos fármacos , Veias Umbilicais/citologia
5.
BMC Microbiol ; 2: 36, 2002 Nov 26.
Artigo em Inglês | MEDLINE | ID: mdl-12453305

RESUMO

BACKGROUND: Chlamydiae are obligate intracellular bacteria, which are important human pathogens. Genome sequences of C. trachomatis and C. pneumoniae have revealed the presence of a Chlamydia specific gene family encoding polymorphic outer membrane proteins, Pmps. In C. pneumoniae the family comprises twenty-one members, which are all transcribed. In the present study, the expression, processing and localisation of the sixteen full-length Pmps in C. pneumoniae strain CWL029 have been further investigated by two-dimensional gel electrophoresis and immunofluorescence microscopy. RESULTS: Ten Pmps were identified in elementary bodies (EBs). Eight of these were investigated with respect to time dependent expression and all were found to be up-regulated between 36 and 48 hours post infection. Antibodies against Pmp6, 8, 10, 11 and 21 reacted with chlamydiae when infected cells were formalin fixed. Pmp6, Pmp20 and Pmp21 were found in cleaved forms, and the cleavage sites of Pmp6 and Pmp21 were identified. CONCLUSIONS: The Pmps are heavily up-regulated at the time of conversion of RB to EB, and at least ten Pmps are present in EBs. Due to their reaction in formalin fixation it is likely that Pmp6, 8, 10, 11 and 21 are surface exposed. The identified cleavage sites of Pmp6 and Pmp21 are in agreement with the theory that the Pmps are autotransporters.


Assuntos
Proteínas da Membrana Bacteriana Externa/metabolismo , Proteínas de Bactérias/metabolismo , Chlamydophila pneumoniae/metabolismo , Proteínas da Membrana Bacteriana Externa/química , Proteínas da Membrana Bacteriana Externa/genética , Proteínas de Bactérias/genética , Proteínas de Bactérias/isolamento & purificação , Chlamydophila pneumoniae/genética , DNA Bacteriano/genética , Eletroforese em Gel de Poliacrilamida , Regulação Bacteriana da Expressão Gênica , Humanos , Espectrometria de Massas , Microscopia de Fluorescência , Reação em Cadeia da Polimerase , Análise de Sequência de Proteína , Células Tumorais Cultivadas
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