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1.
BMC Res Notes ; 15(1): 23, 2022 Jan 28.
Artigo em Inglês | MEDLINE | ID: mdl-35090555

RESUMO

OBJECTIVE: The study of the circadian clock and its mechanisms is easily facilitated through clock resetting in cell culture. Among the various established synchronizers of the circadian clock in cell culture (temperature, serum shock, glucocorticoids), the artificial glucocorticoid Dexamethasone (DEX) is the most widely used. DEX treatment as a protocol to reset the circadian clock in culture gives simple readout with minimal laboratory requirements. Even though there are many studies regarding clock resetting in culture using DEX, reference points or expression patterns of core clock genes and their protein products are scarce and sometimes contradict other works with similar methodology. We synchronise a cell line of human origin with DEX to be used for studies on circadian rhythms. RESULTS: We treat HEK 293T cells with DEX and describe the patterns of mRNA and proteins of core clock regulators, while making a clear point on how CLOCK is less than an ideal molecule to help monitor rhythms in this cell line.


Assuntos
Relógios Circadianos , Proteínas Circadianas Period , Ritmo Circadiano , Dexametasona/farmacologia , Células HEK293 , Humanos , Proteínas Circadianas Period/metabolismo
2.
In Vivo ; 32(1): 109-112, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29275306

RESUMO

BACKGROUND: Human exposure to engineered nanoparticles has been linked to pleural effusion, inflammation and fibrosis. Silver nanoparticles (AgNPs) are widely used in medical and domestic products, increasing the risk of occupational and domestic exposure. We assessed the influence of AgNPs on adhesion and proliferation of sheep primary pleural mesothelial cells. MATERIALS AND METHODS: Cells were used for cell adhesion (90 min) and proliferation experiments (3 days) while exposed to 20 nm and 60 nm AgNPs (0.2 µg/ml and 2 µg/ml) using colorimetric assays. RESULTS: Exposure to 0.2 µg/ml of 20 nm and 60 nm AgNPs significantly increased cell adhesion, while at 2 µg/ml this effect was not elicited. Cell proliferation was significantly increased by both 20 nm and 60 nm AgNPs at 0.2 µg/ml, while at 2 µg/ml this effect was only elicited by the 60 nm AgNPs. CONCLUSION: AgNPs alter the adhesive and proliferative properties of primary pleural mesothelial cells.


Assuntos
Proliferação de Células/efeitos dos fármacos , Células Epiteliais/efeitos dos fármacos , Nanopartículas Metálicas/administração & dosagem , Prata/administração & dosagem , Animais , Adesão Celular/efeitos dos fármacos , Células Cultivadas , Relação Dose-Resposta a Droga , Nanopartículas Metálicas/química , Tamanho da Partícula , Pleura/citologia , Ovinos
3.
Environ Toxicol Pharmacol ; 50: 212-215, 2017 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-28192750

RESUMO

Nanoparticles have been implicated in the development of pleural effusions in exposed factory workers while in experimental animal studies it has been shown that they induce inflammation, fibrosis and carcinogenesis in the pleura. The scope of this study was to investigate the direct effects of silver nanoparticles exposure on the membrane permeability of sheep parietal pleura, of primary sheep pleural cell monolayers and on a human mesothelial cell line. Our findings suggest that acute (30min) exposure increases the pleural permeability ex vivo, while longer (24h) exposure in vivo leads to late decrease of the pleural cell monolayers permeability.


Assuntos
Células Epiteliais/efeitos dos fármacos , Pleura/citologia , Prata/toxicidade , Animais , Células Cultivadas , Células Epiteliais/citologia , Humanos , Nanopartículas Metálicas/química , Permeabilidade , Ovinos , Testes de Toxicidade Aguda
4.
Cytokine ; 69(2): 284-8, 2014 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-24986423

RESUMO

Vascular endothelial growth factor (VEGF), a cytokine that increases vascular permeability to water and proteins and induces angiogenesis, has been implicated in the development of pleural effusions. Inflammatory and malignant pleural effusions are rich in VEGF content while mesothelial cells produce and excrete VEGF. In this report we aimed at investigating by means of electrophysiology the direct effects of VEGF on the parietal and visceral sheep pleura as well as the type of receptors that mediate this effect. Our findings show that VEGF has a direct effect on the pleural mesothelium rendering it more permeable and this effect is mediated through the stimulation of VEGF receptor 2. Our findings shed more light to the role of VEGF in the pathogenesis of pleural effusions and provide functional evidence for a role of VEGFR2 on the pleural mesothelium that has never been studied before.


Assuntos
Pleura/efeitos dos fármacos , Pleura/metabolismo , Fator A de Crescimento do Endotélio Vascular/farmacologia , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/metabolismo , Animais , Impedância Elétrica , Feminino , Técnicas In Vitro , Masculino , Permeabilidade/efeitos dos fármacos , Ovinos , Fatores de Tempo
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