RESUMEN
RNA synthesis can be detected by 5-ethynyl uridine (EU) incorporation and click chemistry. Despite identifying a fundamental functional process, this technique has yet to be widely applied to complex human tissue systems. By incorporating EU into human hair follicle (HF) organs cultured ex vivo, nascent RNA synthesis was detected in situ. EU differentially incorporated across the HF epithelium. Interestingly, RNA synthesis did not correlate with protein synthesis, proliferation or epithelial progenitor cell marker expression. By treating human HFs with the cytotoxic cell cycle inhibitor (R)-CR8, which inhibits transcriptional regulators CDK7 and CDK9, it was further shown that this technique can be used to sensitively detect changes in global RNA synthesis in situ. Together, this work delineates new insights into nascent RNA synthesis within a human (mini)- organ and describes a novel read-out parameter that will enrich future ex vivo human tissue research studies.
Asunto(s)
Química Clic , Folículo Piloso/metabolismo , Microscopía Fluorescente , Técnicas de Cultivo de Órganos , ARN/análisis , Uridina/análogos & derivados , Proliferación Celular , Química Clic/métodos , Regulación de la Expresión Génica , Folículo Piloso/citología , Humanos , Microscopía Fluorescente/métodos , Técnicas de Cultivo de Órganos/métodos , ARN/genética , Transcripción Genética , Uridina/genéticaRESUMEN
The quantification of particle emission factors under controlled laboratory conditions for burning of the following five common tree species found in South East Queensland forests has been studied: Spotted Gum (Corymbia citriodora), Blue Gum (Eucalyptus tereticornis), Bloodwood (Eucalyptus intermedia), Iron Bark (Eucalyptus crebra), and Stringybark (Eucalyptus umbra). The results of the study show that the particle number emission factors and PM2.5 mass emission factors depend on the type of tree and the burning rate. For fast burning conditions, the average particle number emission factors are in the range of 3.3-5.7 x 10(15) particles/kg for woods and 0.5-6.9 x 10(15) particles/kg for leaves and branches, and the PM2.5 emission factors are in the range of 140-210 mg/kg for woods and 450-4700 mg/kg for leaves and branches. For slow burning conditions, the average particle number emission factors are in the range of 2.8-44.8 x 10(13) particles/kg for woods and 0.5-9.3 x 10(13) particles/kg for leaves and branches, and the PM2.5 emissions factors are in the range of 120-480 mg/kg for woods and 3300-4900 mg/kg for leaves and branches.