RESUMEN
Progesterone receptor (PR) signaling is required for mammary gland development and homeostasis. A major bottleneck in studying PR signaling is the lack of sensitive assays to measure and visualize PR pathway activity both quantitatively and spatially. Here, we develop new tools to study PR signaling in human breast epithelial cells. First, we generate optimized Progesterone Responsive Element (PRE)-luciferase constructs and demonstrate that these new reporters are a powerful tool to quantify PR signaling activity across a wide range of progesterone concentrations in two luminal breast cancer cell lines, MCF7 and T47D. We also describe a fluorescent lentiviral PRE-GFP reporter as a novel tool to visualize PR signaling at the single-cell level. Our reporter constructs are sensitive to physiological levels of progesterone. Second, we show that low background signaling, and high levels of PR expression are a prerequisite for robustly measuring PR signaling. Increasing PR expression by transient transfection, stable overexpression in MCF7 or clonal selection in T47D, drastically improves both the dynamic range of luciferase reporter assays, and the induction of endogenous PR target genes as measured by qRT-PCR. We find that the PR signaling response differs per cell line, target gene and hormone concentration used. Taken together, our tools allow a more rationally designed approach for measuring PR signaling in breast epithelial cells.
Asunto(s)
Progesterona , Receptores de Progesterona , Humanos , Transducción de Señal , Células MCF-7 , LuciferasasRESUMEN
Isolation and restraint both aim to protect the patient. Their practice, monitored and assessed, must meet ethical, legal, clinical and organisational requirements. It has its advantages (pacification) and its limits (loss of freedom to make choices about one's care).
Asunto(s)
Trastornos Mentales , Aislamiento de Pacientes , Psiquiatría , Restricción Física , Humanos , Trastornos Mentales/terapiaRESUMEN
Few researchers have investigated the relation of children's sleep problems to their parents' sleep problems. Children with autism have been reported to evidence greater sleep problems than do typically developing children (C. D. Hoffman, D. P. Sweeney, J. E. Gilliam, & M. C. Lopez-Wagner, 2006; P. G. William, L. L. Sears, & A. Allard, 2004). In the present study, parents (N = 106) of children independently diagnosed with autism (4-16 years of age; M= 8.20 years, SD = 2.69 years) reported greater sleep problems for themselves than did parents (N = 168) of typically developing children (4-15 years of age; M = 8.62 years, SD = 3.28 years). Children's sleep problems were related to parents' sleep problems for both groups; in the autism group, children's level of symptomatology was not related to their parents' sleep. The authors suggest areas for further research on the sleep problems of children and their parents, the potential interaction of these problems with children's symptomatic behavior, and the relations of these factors to child, parent, and family functioning.