ABSTRACT
OBJECTIVES: To evaluate the impact of a debate on pharmacy students' perceptions, using online pharmacy practice as the debate topic. METHODS: This is a quasi-experimental interrupted time-series study. A 60 min debate was organized as a lunchtime meeting. A four-category Likert scale questionnaire (fully agree, partially agree, partially disagree, fully disagree) measured the debate participants' level of agreement with 25 statements (main issues associated with online pharmacy) in the pre-phase (before the debate), post-phase 1 (after the debate) and post-phase 2 (6 months after the debate). One hundred and seventy-seven students were recruited (response rate of 100% in the pre-phase and post-phase 1, 31% in post-phase 2). Four questions measured the perceptions of the students on this pedagogical technique. KEY FINDINGS: The overall proportion of respondents in favour of online pharmacy practice showed little variation among the three phases. However, on average (mean ± SD) 43 ± 8% of the respondents changed their opinion, 21 ± 7% reversed their opinion, 22 ± 4% nuanced their opinion and 1 ± 1% radically changed their opinion. Respectively 98% (post-phase 1) and 96% (post-phase 2) of the respondents were of the opinion that debate was a very useful teaching formula in their pharmacist training and 79 and 66% thought debate significantly changed their opinion of the issue. CONCLUSIONS: Few data have been collected on the use of debates as part of healthcare professional training. The impact of a debate on how pharmacy students feel about online pharmacy practice is described.
Subject(s)
Education, Pharmacy/methods , Internet , Pharmaceutical Services/organization & administration , Students, Pharmacy/psychology , Adolescent , Adult , Attitude of Health Personnel , Female , Humans , Male , Perception , Surveys and Questionnaires , Young AdultABSTRACT
In efforts to define mechanisms of transcriptional activation by the orphan nuclear receptor NGFI-B (Nur77), we identified TIF1beta by mass spectrometry within a nuclear protein complex containing NGFI-B. TIF1beta, also known as KAP-1 (KRAB domain-associated protein) or KRIP-1, acts as a transcriptional corepressor for many transcription factors, in particular for the Krüppel-associated box domain-containing zinc finger transcription factors. TIF1beta is also an intrinsic component of two chromatin remodeling and histone deacetylase complexes, the N-CoR1 and nucleosome remodeling and deacetylation complexes. In contrast to these activities, we report that TIF1beta is a coactivator of NGFI-B and that it is as potent as the SRC coactivators in this context. Using pull-down assays and immunoprecipitation, we showed that TIF1beta interacts directly with NGFI-B and with other Nur family members. NGFI-B is an important mediator of hypothalamic corticotropin-releasing hormone (CRH) activation of proopiomelanocortin (POMC) transcription, and TIF1beta enhances transcription mediated through the NGFI-B target, the Nur response element (NurRE). The NurRE binds Nur factor dimers and is responsive to signaling pathways. In keeping with the role of NGFI-B as mediator of CRH signaling, we found that TIF1beta is recruited to the POMC promoter following CRH stimulation and that TIF1beta potentiates CRH and protein kinase A signaling through the NurRE; it acts synergistically with the SRC2 coactivator. However, the actions of TIF1beta and SRC2 were mapped to different NGFI-B AF-1 subdomains. Taken together, these results indicate that TIF1beta is an important coactivator of NGFI-B-dependent transcription.
Subject(s)
DNA-Binding Proteins/metabolism , Receptors, Steroid/metabolism , Repressor Proteins/metabolism , Cell Extracts , Cell Line , Cell Nucleus/metabolism , Corticotropin-Releasing Hormone/metabolism , Cyclic AMP-Dependent Protein Kinases/metabolism , DNA-Binding Proteins/chemistry , Humans , Molecular Weight , Nuclear Receptor Coactivator 2/metabolism , Nuclear Receptor Subfamily 4, Group A, Member 1 , Nuclear Receptor Subfamily 4, Group A, Member 2 , Pro-Opiomelanocortin/genetics , Protein Binding , Protein Structure, Tertiary , Repressor Proteins/chemistry , Response Elements/genetics , Signal Transduction , Transcription Factors/metabolism , Transcription, Genetic , Tripartite Motif-Containing Protein 28ABSTRACT
The retinoblastoma tumor suppressor protein (Rb) is best known as a repressor of genes involved in cell cycle progression. Rb has also been implicated in activation of transcription, in particular by nuclear receptors (NRs) and by differentiation-related transcription factors, but the relevance of this activity is unclear. We show that Rb and the related proteins p107 and p130 enhance the activity of NRs related to NGFI-B (Nur factors) through direct interactions with NGFI-B and SRC-2. Although recruitment of SRC/p160 coactivators to the NGFI-B AF1 domain is independent of Rb, its presence enhances SRC-dependent transcription. Rb potentiation of SRC coactivators is exerted on a subset (Nur factors, hepatocyte nuclear factor-4 (HNF-4), SF-1, and ER) but not all NRs. The levels of Rb-related proteins modulate hormone responsiveness of the NGFI-B-dependent pituitary proopiomelanocortin gene and HNF-4-dependent transcription during enterocyte differentiation. Increased Rb expression upon cell differentiation may promote differentiated functions, at least in part, by potentiation of NR activity.
Subject(s)
Receptors, Cytoplasmic and Nuclear/metabolism , Retinoblastoma Protein/metabolism , Transcription Factors/metabolism , Animals , Base Sequence , Caco-2 Cells , Cell Line , DNA-Binding Proteins/chemistry , DNA-Binding Proteins/genetics , DNA-Binding Proteins/metabolism , Humans , Kinetics , L Cells , Mice , Models, Biological , Multiprotein Complexes , Nuclear Receptor Coactivator 2 , Nuclear Receptor Subfamily 4, Group A, Member 1 , Pro-Opiomelanocortin/genetics , Promoter Regions, Genetic , Protein Structure, Tertiary , RNA, Small Interfering/genetics , Receptors, Cytoplasmic and Nuclear/chemistry , Receptors, Cytoplasmic and Nuclear/genetics , Receptors, Steroid/chemistry , Receptors, Steroid/genetics , Receptors, Steroid/metabolism , Recombinant Proteins/genetics , Recombinant Proteins/metabolism , Retinoblastoma Protein/chemistry , Retinoblastoma Protein/genetics , Transcription Factors/chemistry , Transcription Factors/genetics , Transcription, Genetic , TransfectionABSTRACT
Gene inactivation studies have suggested that the product of the retinoblastoma gene, Rb, is particularly limiting in pituitary pro-opiomelanocortin (POMC)-expressing cell lineages. Indeed, in Rb knock-out mice, these cells develop tumors with high frequency. To understand the implication of limiting Rb expression in these cells, we investigated the action of Rb and its related pocket proteins, p107 and p130, on POMC gene transcription. This led to the identification of the neurogenic basic helix-loop-helix transcription factor, NeuroD1, as a target of Rb action. Rb and to a lesser extent p107, but not p130, enhance NeuroD1-dependent transcription, and this activity appears to depend on direct protein interactions between the Rb pocket and the helix-loop-helix domain of NeuroD1. In vivo, NeuroD is found in a complex that includes Rb and also the orphan nuclear receptor NGFI-B, which mediates corticotropin-releasing hormone activation of POMC transcription. The formation of a similar complex in vitro requires the presence of Rb as a bridge between NeuroD and NGFI-B. In POMC-expressing AtT-20 cells, Rb and p107 are present on the POMC promoter and inhibition of their expression through small interfering RNA decreases POMC mRNA levels. The action of Rb and its related proteins on POMC transcription may contribute to the establishment and/or maintenance of the differentiation phenotype.
