IκB kinase inhibition as a potential treatment of osteoarthritis - results of a clinical proof-of-concept study.

OBJECTIVE: This publication summarizes the clinical development of the compound SAR113945, an IκB kinase inhibitor injected intra-articularly in a slow-release formulation to treat patients with symptomatic osteoarthritis (OA) of the knee. RESULTS: In vitro experiments demonstrated a specific inhibition of the IκB kinase complex. Profiling of SAR113945 on kinases, enzymes and ion channels supported the initiation of a clinical development. Cellular assay systems also revealed an inhibition in the synthesis of interleukin 1ß, tumor necrosis factor α (TNFα) and the prostaglandin E2 (PGE2). In vivo studies demonstrated positive effects of SAR113945 on thermal and mechanical hyperalgesia and even showed superiority in comparison with triamcinolone. Pharmacokinetic measurements showed a sustained release of dissolved SAR113945 locally supporting a comparably high exposure in the knee joint combined with a low systemic exposure. Three phase 1 studies with a dose-escalating design confirmed safety and tolerability of SAR113945. In those studies SAR113945 showed a positive trend on the WOMAC scores. The proof-of-concept or phase 2a study failed to show any effect in the overall group of recruited study participants for the primary endpoint, the WOMAC pain subscore at day 56, but showed a statistically significant difference in a subgroup of patients who had presented with effusion at baseline. CONCLUSION: Inhibiting the NFκB signaling pathway is an attractive method to treat patients with signs and symptoms of OA. The preclinical work and the results of the phase 1 studies appeared promising for a full clinical development, however, the proof-of-concept study failed to show efficacy in a larger patient sample size.

Acrylamide formation and antioxidant level in biscuits related to recipe and baking.

Heated plant foods may contain compounds with adverse health effects (e.g. acrylamide). On the other hand, health-promoting compounds (e.g. antioxidants) have also been identified in such foods. Therefore, a baking experiment with biscuits was carried out to study the potential impact of both acrylamide and antioxidants in that food. Two different wheat flour types - wholemeal (WMF) and white flour type 550 (T550; 0.55% mineral content) - as well as recipe (fat and leavening agent) and thermal input (temperature × time) were changed. Furthermore, the effect of an enzymatic asparagine hydrolysation was tested. Antioxidants were determined with two independent procedures ABTS - (2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonate)) and FRAP-assay (ferric reducing ability of plasma). WMF samples resulted in an unchanged acrylamide level, but in a significantly higher antioxidant concentration when compared with T550 samples (149 and 141 µg kg(-1) acrylamide and 9.1 and 5.1 mmol TE kg(-1) FW ABTS for WMF and T550, respectively). A reduced fat content yielded in an increased volume. Raising agents had no effect on acrylamide levels, but antioxidants were higher in samples with sodium bicarbonate (SBC) than with ammonium bicarbonate (ABC). Thermal input (temperature × time; 150°C × 25 min to 240°C × 9 min) indicated an exponential acrylamide increase especially at higher temperatures (from 75 to 236 µg kg(-1)), whereas antioxidant increase was linear (from 7.0 to 7.7 mmol TE kg(-1) FW, ABTS). FRAP and ABTS values were correlated on a low level, whereas acrylamide content of biscuits was correlated with FRAP and lightness (R (2 )= 0.62 and 0.47, and 0.71 and 0.85 for WMF and T550, respectively). The enzyme asparaginase reduced acrylamide formation by about 50% for both raising agents (SBC and ABC, respectively), whereas antioxidant levels were not affected. An evaluation of recipe variants with low acrylamide and high antioxidants indicated the advantage of wholemeal biscuits.

Clinical prioritisations of healthcare for the aged--professional roles.

BACKGROUND: Although fair distribution of healthcare services for older patients is an important challenge, qualitative research exploring clinicians' considerations in clinical prioritisation within this field is scarce. OBJECTIVES: To explore how clinicians understand their professional role in clinical prioritisations in healthcare services for old patients. DESIGN: A semi-structured interview-guide was employed to interview 45 clinicians working with older patients. The interviews were analysed qualitatively using hermeneutical content analysis. PARTICIPANTS: 20 physicians and 25 nurses working in public hospitals and nursing homes in different parts of Norway. RESULTS AND INTERPRETATIONS: The clinicians struggle with not being able to attend to the comprehensive needs of older patients, and being unfaithful to professional ideals and expectations. There is a tendency towards lowering the standards and narrowing the role of the clinician. This is done in order to secure the vital needs of the patient, but is at the expense of good practice and holistic role modelling. Increased specialisation, advances and increase in medical interventions, economical incentives, organisational structures, and biomedical paradigms, may all contribute to a narrowing of the clinicians' role. CONCLUSION: Distributing healthcare services in a fair way is generally not described as integral to the clinicians' role in clinical prioritisations. If considerations of justice are not included in clinicians' role, it is likely that others will shape major parts of their roles and responsibilities in clinical prioritisations. Fair distribution of healthcare services for older patients is possible only if clinicians accept responsibility in these questions.

In quest of justice? Clinical prioritisation in healthcare for the aged.

BACKGROUND: A fair distribution of healthcare services for older patients is an important challenge, but qualitative research exploring clinicians' consideration in daily clinical prioritisation in healthcare services for the aged is scarce. OBJECTIVES: To explore what kind of criteria, values, and other relevant considerations are important in clinical prioritisations in healthcare services for older patients. DESIGN: A semi-structured interview-guide was used to interview 45 clinicians working with older patients. The interviews were analysed qualitatively using hermeneutical content analysis and template organising style. PARTICIPANTS: 20 physicians and 25 nurses working in public hospitals and nursing homes in different parts of Norway. RESULTS AND INTERPRETATIONS: Important dilemmas relate to under-provision of community care and comprehensive approaches, and over-utilisation of certain specialised services. Overt ageism is generally not reported, but the healthcare services for the aged seem to be inadequate due to more subtle processes, for example, dominating considerations and ideals and operating conditions that do not pay sufficient attention to older patients' needs and considerations of justice. Clinical prioritisations are described as being dominated by adapting traditional biomedical approaches to the operating conditions. Many of the clinicians indicate that there is a potential for improving end of life decisions and for reducing exaggerated use of life-prolonging treatment and hospitalisations. CONCLUSION: The interviews in this study indicate that considerations of justice and patients' perspectives should be given more attention to strike a balance between specialised medical approaches and more general and comprehensive approaches in healthcare services for older patients.

Functional characterization and localization of protein phosphatase type 2C from Paramecium.

We cloned a protein phosphatase 2C gene from Paramecium (PtPP2C), which codes for one of the smallest PP2C isoforms (Klumpp, S., Hanke, C., Donella-Deana, A., Beyer, A., Kellner, R., Pinna, L. A., and Schultz, J. E. (1994) J. Biol. Chem. 269, 32774-32780). After mutation of 9 ciliate Q codons (TAA) to CAA PtPP2C was expressed as an active protein in Escherichia coli. The catalytic core region contains 284 amino acids as defined by C- and N-terminal deletions. The C terminus from amino acid 200-300 of PP2C isoforms has only about 20% similarity. To demonstrate that the carboxy end is in fact needed for activity, we generated an enzymatically active PtPP2C containing a C-terminally located tobacco etch virus-protease site. Upon proteolytic truncation enzyme activity was lost, i.e. the C terminus of PP2C is indispensable for enzyme activity. During these experiments isoleucine 214 was fortuitously identified to be essential for PP2C catalysis. Mutation of the hydrophobic amino acid to glycine in the ciliate or bovine isoforms resulted in inactive protein. Because Ile214 is in a loop region without defined secondary structure, our data clearly go beyond the x-ray structure. The functional equivalence of the 180 amino acid long C terminus from the bovine PP2C with the 100 amino acid long carboxy end of the PtPP2C was demonstrated by producing an active chimera, i.e. the PP2C from Paramecium has no obvious regions which may be specifically involved in subcellular localization or substrate recognition. Using antibodies against recombinant PtPP2C we localized the enzyme by immunogold labeling in the cytosol and nucleus and very distinctly on the ciliary microtubule/dynein complex. The data suggest a role for PtPP2C in the regulation of dyneins, i.e. in cellular cargo transport and ciliary motility.