RESUMEN
Aim: Determining the stability of biomarkers continues to present challenges. Disease states, complex matrices and differences between recombinant and endogenous analytes require new approaches to maintain stability and measure it. In this report, we determine stability for two assays using trending and statistical analysis. Methodology & results: Monitoring trends helps identify out of specification measurements and determine whether concerns are due to the stability of the analyte. We also describe challenges presented when measuring arginase activity in human sputum, a complex matrix, for respiratory diseases. We controlled preanalytical protease activity and collection heterogeneity and monitored incurred sample stability to improve stability of arginine. Conclusion: These new approaches to achieving and determining biomarker stability may provide solutions for increasingly complex biomarker measurements.
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Biomarcadores/análisis , Técnicas de Química Analítica/métodos , Arginasa/química , Arginasa/metabolismo , Biomarcadores/química , Humanos , Subunidad alfa1 del Receptor de Interleucina-13/análisis , Subunidad alfa1 del Receptor de Interleucina-13/química , Estabilidad Proteica , Control de Calidad , Esputo/enzimología , Estadística como Asunto , Factor de Crecimiento Transformador beta1/análisis , Factor de Crecimiento Transformador beta1/químicaRESUMEN
Interleukin (IL)-13-associated signal pathway plays an important role in schistosomiasis hepatic fibrosis. In this study we tried to investigate the effects of corilagin to ameliorate schistosomiasis hepatic fibrosis through regulating IL-13-associated signal pathway in vitro and in vivo. Cellular model was set up with hepatic stellate cells-T6 cells stimulated by rIL-13 and male Balb/c mice were infected with Schistosoma japonicum cercariaeas as animal model. Liver histological changes were observed with haematoxylin and eosin staining. Masson staining was employed to observe the change of egg granulomas. Expression of Col (collagen) and Col III were examined with Immunohistochemistry. Western bolt was employed to detect the JAK-1 and IL13Rα1 proteins. The mRNA expression of Col I, Col III, IL-13, JAK-1 and IL13Rα1 were tested by quantitative polymerase chain reaction. As a result, less inflammatory changes were found in all corilagin groups compared with model group and praziquantel group. The mRNA levels of Col I, Col III, IL-13, JAK-1 and IL13Rα1 were significantly decreased after corilagin intervention (P < 0·01). JAK-1 and IL-13Rα1 protein levels were also greatly decreased in the corilagin groups (P < 0·01). In conclusion, corilagin could ameliorate schistosomiasis hepatic fibrosis by down-regulating the expression of IL-13 and signal molecules in IL-13 pathway.
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Fármacos Gastrointestinales/administración & dosificación , Glucósidos/administración & dosificación , Taninos Hidrolizables/administración & dosificación , Interleucina-13/metabolismo , Cirrosis Hepática/patología , Cirrosis Hepática/terapia , Esquistosomiasis/complicaciones , Transducción de Señal , Animales , Western Blotting , Línea Celular , Colágeno/análisis , Modelos Animales de Enfermedad , Perfilación de la Expresión Génica , Histocitoquímica , Inmunohistoquímica , Subunidad alfa1 del Receptor de Interleucina-13/análisis , Janus Quinasa 1/análisis , Hígado/patología , Ratones Endogámicos BALB C , Microscopía , Modelos Biológicos , Ratas , Reacción en Cadena en Tiempo Real de la Polimerasa , Resultado del TratamientoRESUMEN
BACKGROUND AND AIM OF THE STUDY: Interleukin (IL)-13 is a major inducer of fibrosis in many chronic infectious diseases, yet few studies have reported its role in valvular fibrosis in patients with rheumatic heart disease (RHD). The study aim was to investigate the role of IL-13 in mitral valvular fibrosis in patients with RHD. METHODS: Peripheral blood samples were collected from surgical patients with RHD (n = 18) and from healthy controls (n = 9). Serum levels of IL-13 and interferon (IFN)-gamma were analyzed using ELISA. Rheumatic mitral valves removed from surgical patients with RHD, and normal mitral valves, were obtained at autopsy. The expression and distribution of collagen I, collagen III, and IL-13Ralpha1 were examined by immunohistochemical staining, the degree of which was measured using computed imaging analysis. RESULTS: Higher IL-13 levels were observed in RHD patients (15.16 +/- 9.62 pg/ml; p < 0.05) than in healthy controls (7.78 +/- 3.87 pg/ml). RHD patients had high levels of IFN-gamma (9.95 +/- 0.77 pg/ml; p <0.05) compared to healthy controls (5.95 +/- 0.69 pg/ml). Immunohistochemistry showed that, compared to normal valves, rheumatic mitral valves expressed high levels of collagen I (0.01931 +/- 0.00159 versus 0.01183 +/- 0.00207; p < 0.05), collagen III (0.00726 +/- 0.00078 versus 0.00342 +/- 0.00124; p <0.05), and IL-13Rcxl (0.00454 +/- 0.00086 versus 0.00017 +/- 0.00008; p <0.01). Collagens I and III were each expressed in heart interstitial cells, while IL-13Ralpha1 was expressed in the endothelial cells and smooth muscle cells of the blood vessels, and in interstitial cells. CONCLUSION: Patients with RHD showed increased serum levels of IL-13 compared to healthy controls. IFN-gamma levels were clearly different among RHD patients and healthy controls. The expression of collagens I and III and IL-13Ralpha1 was higher in rheumatic mitral valves compared to normal mitral valves. IL-13 may induce mitral valvular fibrosis in RHD.
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Enfermedades de las Válvulas Cardíacas/diagnóstico , Subunidad alfa1 del Receptor de Interleucina-13/análisis , Interleucina-13/sangre , Válvula Mitral/química , Cardiopatía Reumática/diagnóstico , Adulto , Biomarcadores/sangre , Estudios de Casos y Controles , Colágeno Tipo I/análisis , Colágeno Tipo III/análisis , Ensayo de Inmunoadsorción Enzimática , Femenino , Fibrosis , Enfermedades de las Válvulas Cardíacas/sangre , Enfermedades de las Válvulas Cardíacas/patología , Humanos , Procesamiento de Imagen Asistido por Computador , Inmunohistoquímica , Interferón gamma/sangre , Masculino , Persona de Mediana Edad , Válvula Mitral/patología , Cardiopatía Reumática/sangre , Cardiopatía Reumática/patología , Transducción de Señal , Regulación hacia Arriba , Adulto JovenRESUMEN
The earliest thymic progenitors (ETPs) were recently shown to give rise to both lymphoid and myeloid cells. Whereas the majority of ETPs are derived from IL-7Rα-positive cells and give rise exclusively to T cells, the origin of the myeloid cells remains undefined. In this study, we show both in vitro and in vivo that IL-13Rα1(+) ETPs yield myeloid cells with no potential for maturation into T cells, whereas IL-13Rα1(-) ETPs lack myeloid potential. Moreover, transfer of lineage-negative IL-13Rα1(+) bone marrow stem cells into IL-13Rα1-deficient mice reconstituted thymic IL-13Rα1(+) myeloid ETPs. Myeloid cells or macrophages in the thymus are regarded as phagocytic cells whose function is to clear apoptotic debris generated during T cell development. However, the myeloid cells derived from IL-13Rα1(+) ETPs were found to perform Ag-presenting functions. Thus, IL-13Rα1 defines a new class of myeloid restricted ETPs yielding APCs that could contribute to development of T cells and the control of immunity and autoimmunity.