To our knowledge, the data provide the first indication for the operation of bcl-2 in ventricular myocytes as an antiapoptotic factor.

Introduction: The risks to patient safety and quality of care faced by members of linguistic minority groups have been well-documented. However, little research has focused on the experience of official language minorities in Canada. Methods: This multiple method study (online and paper-based surveys combined with semi-structured individual interviews with patients and interpreters-health navigators) explored the experience of minority Francophones living in 4 Canadian provinces. Results: Patients and interpreters-navigators described experiences where language barriers contributed to poorer patient assessment, misdiagnosis and/or delayed treatment, incomplete understanding of patient condition and prescribed treatment, and impaired confidence in services received. Reliance on Google Translate and ad hoc, untrained interpreters are commonly reported, in spite of evidence highlighting the risks associated with such practice. Conclusion: Increased awareness that the risks of language barriers apply to official language minorities is essential.

Nuclear factor B (NF B) is a ubiquitously expressed transcription factor that is regulated by the cytoplasmic inhibitor protein I B␣. Biological agents such as tumor necrosis factor ␣ (TNF␣), which activate NF B, result in the rapid degradation of I B␣. Adenoviral-mediated gene transfer of Bcl-2 prevents apoptosis of neonatal ventricular myocytes induced by TNF␣. In view of the growing evidence that NF B may play an important role in regulating apoptosis, we determined whether TNF␣ and Bcl-2 could modulate the activity of NF B in ventricular myocytes. Stimulation of myocytes with TNF␣ resulted in a 2.1-fold increase (p < 0.001) in NF B-dependent gene transcription and nuclear DNA binding. Similarly, a 1.9-fold increase (p < 0.0002) in NF B-dependent gene transcription was observed in myocytes expressing Bcl-2. Nuclear DNA binding activity of NF B was significantly increased in myocytes expressing Bcl-2, with a concomitant reduction in I B␣ protein level. The Bcl-2-mediated loss of I B␣ could be prevented by the proteasome inhibitor lactacystin, consistent with the notion that the targeted degradation of I B␣ consequent to overexpression of Bcl-2 utilizes the ubiquitin-proteasome pathway. This was further tested in human 293 cells in which the N-terminal region of I B␣ was identified to be an important regulatory site for Bcl-2. Deletion of this region or a serine to alanine substitution mutant at amino acids 32 and 36, which are defective for both phosphorylation and degradation, were more resistant than wild type I B␣ to the inhibitory effects of Bcl-2. To our knowledge, this provides the first evidence for the regulation of I B␣ by Bcl-2 and suggests a link between Bcl-2 and the NF B signaling pathway in the suppression of apoptosis.The nuclear factor B (NF B) 1 was first identified as a key regulatory molecule necessary for the activation of B lymphocytes gene transcription (1, 2). Because of these initial observations, it is now widely appreciated that NF B is a ubiquitously expressed transcription factor involved in the activation of genes associated with inflammation, cell adhesion, and viral gene transcription (reviewed in Refs. 3 and 4). NF B belongs to a family of transcription factors with Rel homology and include Rel-A, c-Rel, RelB, and Drosophilia dorsal proteins (5-7). The predominant form of NF B exists in mammalian cells as a heterodimeric complex of 50-kDa and 65-kDa/ RelA protein subunits (8 -10). NF B activity can be induced in a number of cell types by a variety of agents, including ionizing radiation, phorbol esters, and proinflammatory cytokines such as interleukin-1 and tumor necrosis factor alpha (TNF␣) (11,12).In contrast to other transcription factors that are typically located within the nucleus of the cell, NF B is sequestered in the cytoplasm by the inhibitor protein I B␣ (5, 13-15). I B␣ prevents the nuclear targeting of NF B by interaction via its conserved ankyrin repeats (7,16,17).The mechanism by which biological signals activate NF B in vivo remains elusive; however, recent stud...

Nuclear factor-kappa B (NF-kappa B) is a ubiquitously expressed cellular factor regulated by the cytoplasmic factor inhibitor protein kappa B alpha (I kappa B alpha). Activation of NF-kappa B by cytokines, including tumor necrosis factor-alpha (TNF-alpha), requires the phosphorylation and degradation of I kappa B alpha. An anti-apoptotic role for NF-kappa B has recently been suggested. In the present study, we ascertained whether death-promoting signals and apoptosis mediated by TNF-alpha are suppressed by NF-kappa B in postnatal ventricular myocytes. Stimulation of myocytes with TNF-alpha resulted in a 12.1-fold increase (P < 0.01) in NF-kappa B-dependent gene transcription and DNA binding compared with controls. This was accompanied by a corresponding increase in the NF-kappa B target protein A20 as determined by Western blot analysis. Vital staining revealed that TNF-alpha was not cytotoxic to myocytes and did not provoke apoptosis. Adenovirus-mediated delivery of a nonphosphorylatable form of I kappa B alpha to inactivate NF-kappa B prevented TNF-alpha-stimulated NF-kappa B-dependent gene transcription and nuclear NF-kappa B DNA binding. Importantly, myocytes stimulated with TNF-alpha and defective for NF-kappa B activation resulted in a 2.2-fold increase (P < 0.001) in apoptosis. To our knowledge, the data provide the first indication that a functional NF-kappa B signaling pathway is crucial for suppressing death-promoting signals mediated by TNF-alpha in ventricular myocytes.

Background: Emerging evidence that meaningful relationships with knowledge users are a key predictor of research use has led to promotion of partnership approaches to health research. However, little is known about health system experiences of collaborations with university-based researchers, particularly with research partnerships in the area of health system design and health service organization. The purpose of the study was to explore the experience and perspectives of senior health managers in health service organizations, with health organization-university research partnerships.Methods: In-depth, semi-structured interviews (n = 25) were conducted with senior health personnel across Canada to explore their perspectives on health system research; experiences with health organization-university research partnerships; challenges to partnership research; and suggested actions for improving engagement with knowledge users and promoting research utilization. Participants, recruited from organizations with regional responsibilities, were responsible for system-wide planning and support functions.Results: Research is often experienced as unhelpful or irrelevant to decision-making by many within the system. Research, quality improvement (QI) and evaluation are often viewed as separate activities and coordinated by different responsibility areas. Perspectives of senior managers on barriers to partnership differed from those identified in the literature: organizational stress and restructuring, and limitations in readiness of researchers to work in the fast-paced healthcare environment, were identified as major barriers. Although the need for strong executive leadership was emphasized, "multi-system action" is needed for effective partnerships.Conclusion: Common approaches to research and knowledge translation are often not appropriate for addressing issues of health service design and health services organization. Nor is the research community providing expertise to many important activities that the healthcare system is taking to improve health services. A radical rethinking of how we prepare health service researchers; position research within the health system; and fund research activities and infrastructure is needed if the potential benefits of research are to be achieved. Lack of response to health system needs may contribute to research and ‘evidence-informed’ practice being further marginalized from healthcare operations. Interventions to address barriers must respond to the perspectives and experience of health leadership.

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Nuclear factor (NF)B

The interactions of the phosphorylated derivatives of hydroquinone (HQN-P2), resorcinol (RSN-P2), 4-hydroxybenzaldehyde (HBA-P) and 2, 4-dihydroxybenzaldehyde (DHBA-P; phosphate group at position 4) with fructose bisphosphate aldolase were analysed by enzyme kinetics, UV/visible difference spectroscopy and site-directed mutagenesis. Enzyme activity was competitively inhibited in the presence of HQN-P2, RSN-P2 and HBA-P, whereas DHBA-P exhibited slow-binding inhibition. Inhibition by DHBA-P involved active-site Schiff-base formation and required a phenol group ortho to the aldehyde moiety. Rates of enzyme inactivation and of Schiff-base formation by DHBA-P were identical, and corresponded to 3.2-3.5 DHBA-P molecules covalently bound per aldolase tetramer at maximal inactivation. Site-directed mutagenesis of the active-site lysine residues at positions 107, 146 and 229 was found to be consistent with Schiff-base formation between DHBA-P and Lys-146, and this was promoted by Lys-229. Mutation of Glu-187, located vicinally between Lys-146 and Lys-229 in the active site, perturbed the rate of Schiff-base formation, suggesting a functional role for Glu-187 in Schiff-base formation and stabilization. The decreased cleavage activity of the active-site mutants towards fructose 1, 6-bisphosphate is consistent with a proton-transfer mechanism involving Lys-229, Glu-187 and Lys-146.