ABSTRACT
Cushing's syndrome (CS) is a rare endocrine disease, due to cortisol hypersecretion. CS patients have comorbidities, often still present after biochemical cure. Specific nursing healthcare programs to address this disease and achieve improved health related quality of life (HRQoL) are lacking. Thus, an educational nursing intervention, through the development and promotion of specific educational tools, appears to be justified. The objective of this study is to assess the effectiveness of an educational nursing program in CS patients on HRQoL, clinical parameters, level of pain and physical activity, patterns of rest, and use of health resources. A prospective, randomized study was conducted in two reference hospitals for CS. Sixty-one patients (mean age 47 ± 12.7 years, 83.6 % females) were enrolled and divided into 2 groups: an "intervention" group where educational sessions were performed over 9 months and a "control" group, without these sessions. Specific questionnaires were used at the beginning and end of the study. After educational sessions, the intervention group had a better score in the CushingQoL questionnaire (p < 0.01), reduced level of pain (p < 0.05), improved physical activity (p < 0.01) and healthy lifestyle (p < 0.001) compared to the control group. A correlation between the CushingQoL score and reduced pain (r = 0.46, p < 0.05), improved physical activity (r = 0.89, p < 0.01), and sleep (r = 0.53, p = 0.01) was observed. This educational nursing program improved physical activity, healthy lifestyle, better sleep patterns, and reduced pain in CS patients, influencing HRQoL and reducing consumption of health resources. Moreover, the brief nature of the program suggests it as a good candidate to be used in CS patients.
Subject(s)
Cushing Syndrome/nursing , Education, Nursing , Quality of Life/psychology , Adult , Exercise , Female , Health Status , Humans , Life Style , Male , Middle Aged , Pain Measurement , Prospective Studies , Sleep , Surveys and QuestionnairesABSTRACT
Understanding how hematopoietic stem cells (HSCs) are generated and the signals that control this process is a crucial issue for regenerative medicine applications that require in vitro production of HSC. HSCs emerge during embryonic life from an endothelial-like cell population that resides in the aorta-gonad-mesonephros (AGM) region. We show here that ß-catenin is nuclear and active in few endothelial nonhematopoietic cells closely associated with the emerging hematopoietic clusters of the embryonic aorta during mouse development. Importantly, Wnt/ß-catenin activity is transiently required in the AGM to generate long-term HSCs and to produce hematopoietic cells in vitro from AGM endothelial precursors. Genetic deletion of ß-catenin from the embryonic endothelium stage (using VE-cadherin-Cre recombinase), but not from embryonic hematopoietic cells (using Vav1-Cre), precludes progression of mutant cells toward the hematopoietic lineage; however, these mutant cells still contribute to the adult endothelium. Together, those findings indicate that Wnt/ß-catenin activity is needed for the emergence but not the maintenance of HSCs in mouse embryos.
Subject(s)
Hematopoietic Stem Cells/cytology , Hematopoietic Stem Cells/metabolism , Wnt Proteins/metabolism , beta Catenin/metabolism , Animals , Aorta/embryology , Aorta/growth & development , Aorta/metabolism , Cell Differentiation/physiology , Endothelial Cells/metabolism , Gonads/embryology , Gonads/growth & development , Gonads/metabolism , Hematopoietic Stem Cell Transplantation/methods , Mesonephros/embryology , Mesonephros/growth & development , Mesonephros/metabolism , Mice , Mice, Inbred C57BL , Mutation/genetics , Wnt Proteins/genetics , beta Catenin/geneticsABSTRACT
14-3-3σ is frequently lost in human breast cancers by genetic deletion or promoter methylation. We have now investigated the involvement of 14-3-3σ in the termination of NF-κB signal in mammary cells and its putative role in cancer relapse and metastasis. Our results show that 14-3-3σ regulates nuclear export of p65-NF-κB following chronic TNFα stimulation. Restoration of 14-3-3σ in breast cancer cells reduces migration capacity and metastatic abilities in vivo. By microarray analysis, we have identified a genetic signature that responds to TNFα in a 14-3-3σ-dependent manner and significantly associates with different breast and other types of cancer. By interrogating public databases, we have found that over-expression of this signature correlates with poor relapse-free survival in breast cancer patients. Finally, screening of 96 human breast tumors showed that NF-κB activation strictly correlates with the absence of 14-3-3σ and it is significantly associated with worse prognosis in the multivariate analysis. Our findings identify a genetic signature that is important for breast cancer prognosis and for future personalized treatments based on NF-κB targeting.
Subject(s)
14-3-3 Proteins/metabolism , Breast Neoplasms/metabolism , NF-kappa B/antagonists & inhibitors , Tumor Suppressor Proteins/metabolism , 14-3-3 Proteins/genetics , Active Transport, Cell Nucleus , Animals , Breast Neoplasms/genetics , Breast Neoplasms/mortality , Cell Line, Tumor , Cell Movement/genetics , Cluster Analysis , Enzyme Activation/drug effects , Female , Gene Expression , Gene Expression Profiling , Humans , Mice , NF-kappa B/metabolism , Neoplasm Metastasis , Prognosis , Protein Binding/drug effects , Survival Analysis , Transcription, Genetic , Tumor Necrosis Factor-alpha/pharmacology , Tumor Suppressor Proteins/geneticsABSTRACT
It was previously shown that the NF-κB pathway is downstream of oncogenic Notch1 in T cell acute lymphoblastic leukemia (T-ALL). Here, we visualize Notch-induced NF-κB activation using both human T-ALL cell lines and animal models. We demonstrate that Hes1, a canonical Notch target and transcriptional repressor, is responsible for sustaining IKK activation in T-ALL. Hes1 exerts its effects by repressing the deubiquitinase CYLD, a negative IKK complex regulator. CYLD expression was found to be significantly suppressed in primary T-ALL. Finally, we demonstrate that IKK inhibition is a promising option for the targeted therapy of T-ALL as specific suppression of IKK expression and function affected both the survival of human T-ALL cells and the maintenance of the disease in vivo.
Subject(s)
Basic Helix-Loop-Helix Transcription Factors/metabolism , Homeodomain Proteins/metabolism , Leukemia, T-Cell/metabolism , NF-kappa B/metabolism , Receptors, Notch/metabolism , Tumor Suppressor Proteins/antagonists & inhibitors , Animals , Basic Helix-Loop-Helix Transcription Factors/genetics , Cell Differentiation/physiology , Cell Growth Processes/physiology , Cell Survival/physiology , Deubiquitinating Enzyme CYLD , Genes, Tumor Suppressor , Homeodomain Proteins/genetics , Humans , Leukemia, T-Cell/genetics , Leukemia, T-Cell/pathology , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , Mice, Knockout , NF-kappa B/genetics , Receptors, Notch/genetics , Signal Transduction , Transcription Factor HES-1 , Transcription Factor RelA/metabolism , Tumor Suppressor Proteins/genetics , Tumor Suppressor Proteins/metabolismABSTRACT
Specific deletion of Notch1 and RBPjkappa in the mouse results in abrogation of definitive haematopoiesis concomitant with the loss of arterial identity at embryonic stage. As prior arterial determination is likely to be required for the generation of embryonic haematopoiesis, it is difficult to establish the specific haematopoietic role of Notch in these mutants. By analysing different Notch-ligand-null embryos, we now show that Jagged1 is not required for the establishment of the arterial fate but it is required for the correct execution of the definitive haematopoietic programme, including expression of GATA2 in the dorsal aorta. Moreover, successful haematopoietic rescue of the Jagged1-null AGM cells was obtained by culturing them with Jagged1-expressing stromal cells or by lentiviral-mediated transduction of the GATA2 gene. Taken together, our results indicate that Jagged1-mediated activation of Notch1 is responsible for regulating GATA2 expression in the AGM, which in turn is essential for definitive haematopoiesis in the mouse.
Subject(s)
Aorta/embryology , Calcium-Binding Proteins/metabolism , Hematopoiesis , Intercellular Signaling Peptides and Proteins/metabolism , Membrane Proteins/metabolism , Animals , Calcium-Binding Proteins/genetics , Embryo, Mammalian/metabolism , GATA2 Transcription Factor/metabolism , Hematopoietic Stem Cells/metabolism , Intercellular Signaling Peptides and Proteins/genetics , Jagged-1 Protein , Jagged-2 Protein , Membrane Proteins/genetics , Mice , Mice, Inbred C57BL , Mice, Inbred Strains , Mutation , Serrate-Jagged ProteinsABSTRACT
Objetivo. Identificar las condiciones laborales de los graduados de la Escuela Universitaria de Enfermería de Sant Pau a los 4 años de la graduación y comparar esta situación con la que tenían a los 7 meses de ésta. Identificar los elementos que han favorecido la contratación laboral y comprobar si han logrado emanciparse. Método. Diseño descriptivo, observacional y longitudinal. Se compara la situación laboral a los 4 años (fase 2 [F2]) de la graduación, de 62 enfermeras graduadas el año 2000, con la que tenían a los 7 meses (fase 1 [F1]) de ésta. Instrumento: cuestionario semiestructurado cumplimentado mediante entrevista. Resultados. A los 4 años de graduarse trabajaba el 95%, mientras que a los 7 meses lo hacía el 82%. La media de contratos firmados fue de 7 en F1 y de 87 en F2. No hay diferencias respecto a los principales factores que favorecen la ocupación: "disponibilidad para cualquier turno" (F2: 69%; F1: 65%) y "preparación teórica y práctica" (F2: 53%; F1: 57%). Presentarse personalmente en los centros sigue siendo la mejor estrategia para conseguir trabajo (F2: 85%; F1: 79%). El 48% sigue sin emanciparse. Conclusiones. A los 4 años de la graduación, la inserción laboral es casi completa, pero las condiciones de trabajo siguen siendo inestables y precarias, y comparativamente a los 7 meses ésta ha empeorado. Los elementos que más favorecen la contratación laboral son la disponibilidad horaria y la preparación académica. Sólo la mitad de los graduados ha conseguido emanciparse
Objective. To identify the working conditions of Sant Pau Nursing School graduates 4 years after graduation, and to compare these conditions with those at 7 months after graduation. To determine the elements favoring employment and to quantify graduates who had gained independence from the home. Method. We performed an observational, descriptive, longitudinal study in two phases. Working conditions at 4 years after graduation (phase 2) were compared with those at 7 months after graduation (phase 1). The instrument used was a semi-structured questionnaire completed in an individual interview. Subjects consisted of 62 nurses who graduated in 2000. Results. Four years after graduation, 95% of the nurses were working compared with 82% at 7 months after graduation. The mean number of jobs held was seven for phase 1 and 89 for phase 2. No differences were observed between phases 1 and 2 in the main factors favoring employment: "availability for any shift" (phase 1: 65%, phase 2: 69%;), and "theoretical and practical training" (phase 1: 57%, phase 2: 53%;). The most effective strategy for finding a job continued to be visiting a center. Forty-eight percent continued to live at home. Conclusions. Four years after graduation, almost all the graduates are employed, but working conditions such as job stability and security remain poor. The situation at phase 2 was worse than that at phase F1. Factors favoring employment are hours of availability and the nursing school attended. Only half of the graduates have been able to leave home
Subject(s)
Humans , Nursing Staff/statistics & numerical data , 16360 , Career Mobility , Achievement , Job SatisfactionABSTRACT
IkappaB are responsible for maintaining p65 in the cytoplasm under non-stimulating conditions and promoting the active export of p65 from the nucleus following NFkappaB activation to terminate the signal. We now show that 14-3-3 proteins regulate the NFkappaB signaling pathway by physically interacting with p65 and IkappaBalpha proteins. We identify two functional 14-3-3 binding domains in the p65 protein involving residues 38-44 and 278-283, and map the interaction region of IkappaBalpha in residues 60-65. Mutation of these 14-3-3 binding domains in p65 or IkappaBalpha results in a predominantly nuclear distribution of both proteins. TNFalpha treatment promotes recruitment of 14-3-3 and IkappaBalpha to NFkappaB-dependent promoters and enhances the binding of 14-3-3 to p65. Disrupting 14-3-3 activity by transfection with a dominant-negative 14-3-3 leads to the accumulation of nuclear p65-IkappaBalpha complexes and the constitutive association of p65 with the chromatin. In this situation, NFkappaB-dependent genes become unresponsive to TNFalpha stimulation. Together our results indicate that 14-3-3 proteins facilitate the nuclear export of IkappaBalpha-p65 complexes and are required for the appropriate regulation of NFkappaB signaling.
Subject(s)
14-3-3 Proteins/metabolism , Active Transport, Cell Nucleus/physiology , I-kappa B Proteins/metabolism , Transcription Factor RelA/metabolism , 14-3-3 Proteins/genetics , Amino Acid Sequence , Animals , Cell Line , Chromatin/metabolism , Humans , I-kappa B Proteins/genetics , Mice , Mice, Knockout , Multiprotein Complexes , NF-KappaB Inhibitor alpha , Protein Binding , RNA, Small Interfering/genetics , RNA, Small Interfering/metabolism , Sequence Alignment , Signal Transduction/physiology , Transcription Factor RelA/genetics , Transcription, Genetic , Tumor Necrosis Factor-alpha/metabolismABSTRACT
Objetivo. Describir la situación laboral de los graduados, conocer las causas de desocupación, explicar las causas de abandono profesional, conocer los elementos que favorecen la ocupación, identificar la adecuación de los estudios respecto a la práctica profesional y ver si existe relación entre el tipo de contrato laboral y la independencia del hogar familiar.Diseño. Estudio observacional-longitudinal.Instrumento. Cuestionario semiestructurado cumplimentado mediante entrevista presencial.Emplazamiento. Escuela Universitaria de Enfermería.Sujetos de estudio. Graduados del año 2000.Resultados principales. Se ha entrevistado a 70 graduados. El 86,6 por ciento había encontrado trabajo al mes de haberse graduado, el 95,5 por ciento firmó contratos temporales, el 40,4 por ciento firmó entre 6 y 10 contratos, y el 52,5 por ciento, contratos de un día. Para el 61,4 por ciento, tener disponibilidad para cualquier turno y horario es lo que más contribuye a conseguir trabajo. Para el 55,95 por ciento, la relación entre la formación recibida y los requerimientos del puesto de trabajo está entre el 50 y el 70 por ciento. El 82,9 por ciento manifestó vivir todavía con los padres.Conclusiones. La inserción laboral de nuestros graduados es muy elevada y el período que tardan en encontrar el primer trabajo es muy corto. La movilidad, la polivalencia exigida y la duración de los contratos evidencian unas condiciones laborales precarias que, a su vez, dificultan la adquisición de la cualidad de experto profesional. En el momento de la recogida de los datos no existe abandono de la profesión. La realización de prácticas clínicas ha sido un elemento favorecedor de la inserción laboral. La diferencia de adecuación entre la formación recibida y los requerimientos del lugar de trabajo aumenta cuando el recién graduado trabaja en áreas de mayor especialización. No se observa relación entre el tipo de contrato laboral y la independencia económica y del hogar familiar (AU)
Subject(s)
Adult , Female , Male , Humans , Nursing/statistics & numerical data , Work/statistics & numerical data , Job Application , Longitudinal Studies , Education, Nursing/statistics & numerical data , 16360 , Job Satisfaction , Nurses/statistics & numerical dataABSTRACT
Phosphorylation of Notch proteins has been indirectly correlated with Notch activation and nuclear translocation as well as cellular transformation. There is evidence that the Wnt signaling pathway, which results in glycogen synthase kinase-3 beta (GSK-3 beta) inhibition, cross-talks with the Notch pathway. In this study, we show that GSK-3 beta is able to bind and phosphorylate Notch2 in vitro and in vivo. We identify three specific phosphorylation sites in the Notch2 serine/threonine-rich domain that are dependent on GSK-3 beta activity. Phosphorylation of the serine/threonine-rich domain has been shown previously to be crucial in regulating cytokine-specific cell differentiation. Coimmunoprecipitation experiments show that full-length Notch2 binds more efficiently than intracellular Notch2 to GSK-3 beta. Nevertheless, only the processed Notch2 is a substrate for the kinase, thus suggesting that GSK-3 beta-dependent phosphorylation may be specifically regulating the activated Notch molecule. Consistent with this, GSK-3 beta inhibits the transcriptional activation of Notch target genes both in vitro and in vivo, whereas lithium chloride treatment or Wnt-1 overexpression that results in GSK-3 beta inhibition leads to the up-regulation of the Hes-1 promoter. Together, our results suggest that cross-talk between Notch and Wnt pathways may be partially mediated by specific regulation of GSK-3 beta-dependent Notch phosphorylation.
Subject(s)
Down-Regulation , Glycogen Synthase Kinase 3/metabolism , Proto-Oncogene Proteins/metabolism , Receptors, Cell Surface/metabolism , Zebrafish Proteins , Ankyrin Repeat , Base Sequence , DNA Primers , Glycogen Synthase Kinase 3 beta , Phosphorylation , Promoter Regions, Genetic , Receptor, Notch2 , Receptors, Cell Surface/antagonists & inhibitors , Saccharomyces cerevisiae Proteins/metabolism , Subcellular Fractions/enzymology , Wnt Proteins , Wnt1 ProteinABSTRACT
Notch and NFkappaB pathways are key regulators of numerous cellular events such as proliferation, differentiation, or apoptosis. In both pathways, association of effector proteins with nuclear corepressors is responsible for their negative regulation. We have previously described that expression of a p65-NFkappaB mutant that lacks the transactivation domain (p65DeltaTA) induces cytoplasmic translocation of N-CoR leading to a positive regulation of different promoters. Now, we show that cytoplasmic sequestration of p65 by IkappaBalpha is sufficient to both translocate nuclear corepressors SMRT/N-CoR to the cytoplasm and upregulate transcription of Notch-dependent genes. Moreover, p65 and IkappaBalpha are able to directly bind SMRT, and this interaction can be inhibited in a dose-dependent manner by the CREB binding protein (CBP) coactivator and after TNF-alpha treatment, suggesting that p65 acetylation is modulating this interaction. In agreement with this, TNF-alpha treatment results in downregulation of the Hes1 gene. Finally, we present evidence on how this mechanism may influence cell differentiation in the 32D myeloid progenitor system.
Subject(s)
Calcium-Binding Proteins , Cell Nucleus/metabolism , Cytoplasm/metabolism , I-kappa B Proteins/physiology , Membrane Glycoproteins/physiology , Membrane Proteins/metabolism , NF-kappa B/metabolism , Nerve Tissue Proteins/physiology , 3T3 Cells , Active Transport, Cell Nucleus , Animals , Apoptosis , Binding, Competitive , Blotting, Northern , Blotting, Western , Cell Differentiation , Cell Division , Cell Line , Dose-Response Relationship, Drug , Down-Regulation , Flow Cytometry , Humans , I-kappa B Proteins/metabolism , Luciferases/metabolism , Mice , Microscopy, Fluorescence , NF-KappaB Inhibitor alpha , Plasmids/metabolism , Precipitin Tests , Protein Binding , Protein Structure, Tertiary , Receptors, Notch , Synaptotagmin I , Synaptotagmins , Transcriptional Activation , Tumor Necrosis Factor-alpha/metabolism , Up-RegulationABSTRACT
Notch/RBP-Jkappa and nuclear factor-kappaB (NFkappaB) complexes are key mediators of the progression of many cellular events through the activation of specific target gene transcription. Independent observations have shown that activation of Notch-dependent transcription generally correlates with inhibition of differentiation. In contrast, activated NFkappaB complexes are required for progression of differentiation in several systems. Although some interactions between both pathways have been observed, the physiological significance of their connection is unclear. We have now demonstrated that the increase in p65-NFkappaB protein levels enhances Notch-mediated activation of the Hes1 promoter up to three-fold. This effect does not require NFkappaB transcriptional activity, and it is independent of the previously described interaction between Notch and p50-NFkappaB. Furthermore, we show that p65-NFkappaB can modulate subcellular localization of the transcriptional corepressor N-CoR, abrogating N-CoR mediated repression of the Hes1 promoter. In addition, p65-NFkappaB is able to upregulate not only the Hes1 but also other promoters containing SRE and AP-1 sites, which are repressed by N-CoR. Thus, we conclude that p65-NFkappaB can regulate gene expression by a general mechanism that involves cytoplasmic translocation of the transcriptional corepressor protein N-CoR.
