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1.
Microbiol Immunol ; 63(7): 261-268, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31209918

RESUMO

Pasteurella multocida is one of the most important bacteria responsible for diseases of animals. Crude extracts from sonicated P. multocida strain Dainai-1, which is serotype A isolated from bovine pneumonia, were found to inhibit proliferation of mouse spleen cells stimulated with Con A. The crude extract was purified by cation and anion exchange chromatography and hydroxyapatite chromatography. Its molecular weight was 27 kDa by SDS-PAGE and it was named PM27. PM27 was found to inhibit proliferation of mouse spleen cells stimulated with Con A as effectively as did the crude extract; however, its activity was lost after heating to 100°C for 20 min. PM27 did not directly inhibit proliferation of HT-2 cells, which are an IL-2-dependent T cell line, nor did it modify IL-2 production by Con A-stimulated mouse spleen cells. The N-terminal amino acid sequence of PM27 was determined and BLAST analysis revealed its identity to uridine phosphorylase (UPase) from P. multocida. UPase gene from P. multocida Dainai-1 was cloned into expression vector pQE-60 in Escherichia coli XL-1 Blue. Recombinant UPase (rUPase) tagged with His at the C-terminal amino acid was purified with Ni affinity chromatography. rUPase was found to inhibit proliferation of mouse spleen cells stimulated with Con A; however, as was true for PM27, its activity was lost after heating to 100°C for 20 min. Thus, PM27/UPase purified from P. multocida has significant antiproliferative activity against Con A-stimulated mouse spleen cells and may be a virulence factor.


Assuntos
Proteínas da Membrana Bacteriana Externa/farmacologia , Proliferação de Células/efeitos dos fármacos , Pasteurella multocida/metabolismo , Uridina Fosforilase/isolamento & purificação , Uridina Fosforilase/farmacologia , Sequência de Aminoácidos , Animais , Proteínas da Membrana Bacteriana Externa/genética , Proteínas da Membrana Bacteriana Externa/metabolismo , Bovinos , Linhagem Celular/efeitos dos fármacos , Escherichia coli/genética , Humanos , Interleucina-2/metabolismo , Camundongos , Peso Molecular , Pasteurella multocida/genética , Fosforilases , Proteínas Recombinantes , Baço , Linfócitos T/efeitos dos fármacos , Uridina Fosforilase/genética , Uridina Fosforilase/metabolismo
2.
PLoS One ; 10(12): e0143880, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26623647

RESUMO

Homeostatic control of blood glucose is regulated by a complex feedback loop between glucose and insulin, of which failure leads to diabetes mellitus. However, physiological and pathological nature of the feedback loop is not fully understood. We made a mathematical model of the feedback loop between glucose and insulin using time course of blood glucose and insulin during consecutive hyperglycemic and hyperinsulinemic-euglycemic clamps in 113 subjects with variety of glucose tolerance including normal glucose tolerance (NGT), impaired glucose tolerance (IGT) and type 2 diabetes mellitus (T2DM). We analyzed the correlation of the parameters in the model with the progression of glucose intolerance and the conserved relationship between parameters. The model parameters of insulin sensitivity and insulin secretion significantly declined from NGT to IGT, and from IGT to T2DM, respectively, consistent with previous clinical observations. Importantly, insulin clearance, an insulin degradation rate, significantly declined from NGT, IGT to T2DM along the progression of glucose intolerance in the mathematical model. Insulin clearance was positively correlated with a product of insulin sensitivity and secretion assessed by the clamp analysis or determined with the mathematical model. Insulin clearance was correlated negatively with postprandial glucose at 2h after oral glucose tolerance test. We also inferred a square-law between the rate constant of insulin clearance and a product of rate constants of insulin sensitivity and secretion in the model, which is also conserved among NGT, IGT and T2DM subjects. Insulin clearance shows a conserved relationship with the capacity of glucose disposal among the NGT, IGT and T2DM subjects. The decrease of insulin clearance predicts the progression of glucose intolerance.


Assuntos
Intolerância à Glucose/fisiopatologia , Glucose/metabolismo , Insulina/metabolismo , Adulto , Glicemia/metabolismo , Diabetes Mellitus Tipo 2/metabolismo , Diabetes Mellitus Tipo 2/fisiopatologia , Progressão da Doença , Jejum/metabolismo , Jejum/fisiologia , Feminino , Técnica Clamp de Glucose/métodos , Intolerância à Glucose/metabolismo , Teste de Tolerância a Glucose/métodos , Humanos , Resistência à Insulina/fisiologia , Masculino , Pessoa de Meia-Idade , Período Pós-Prandial/fisiologia
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