Rosiglitazone induces the unfolded protein response, but has no significant effect on cell viability, in monocytic and vascular smooth muscle cells.

Given the safety concerns expressed over negative cardiovascular outcomes resulting from the clinical use of rosiglitazone, and the view that rosiglitazone exerts PPARγ-independent effects alongside its insulin-sensitising PPARγ-dependent effects, we hypothesised that rosiglitazone may trigger Unfolded Protein Responses (UPRs) due to disruptions in [Ca(2+)](i) homeostasis within two cardiovascular cell types: monocytic (MM6) and vascular smooth muscle (A7r5) cells. In microsomal samples derived from both cell types, pre-incubation with rosiglitazone rapidly (30min) brought about concentration-dependent PPARγ-independent inhibition of Ca(2+)ATPase activity (IC(50) ∼2µM). Fluo-3 fluorimetric data demonstrated in intact cells that 1h treatment with 1 or 10µM rosiglitazone caused Ca(2+) ions to leak into the cytoplasm. Gene expression analysis showed that within 4h of rosiglitazone exposure, the UPR transcription factor XBP-1 was activated (likely due to corresponding ER Ca(2+) depletion), and the UPR target genes BiP and SERCA2b were subsequently upregulated within 24-72h. After 72h 1 or 10µM rosiglitazone treatment, microsomal Ca(2+)ATPase activity increased to >2-fold of that seen in control microsomes, while [Ca(2+)](i) returned to basal, indicating that UPR-triggered SERCA2b upregulation was responsible for enhanced enzymatic Ca(2+) sequestration within the ER. This appeared to be sufficient to replenish ER Ca(2+) stores and restore normal cell physiology, as cell viability levels were not decreased due to rosiglitazone treatment throughout a 2-week study. Thus, incubation with 1-10µM rosiglitazone triggers the UPR, but does not prove cytotoxic, in cells of the cardiovascular system. This observation provides an important contribution to the current debate over the use of rosiglitazone in the clinical treatment of Type-2 Diabetes.

Rosiglitazone transiently disturbs calcium homeostasis in monocytic cells.

The PPARgamma agonist Rosiglitazone exerts anti-hyperglycaemic effects by regulating the long-term expression of genes involved in metabolism, differentiation and inflammation. In the present study, Rosiglitazone treatment rapidly inhibited (5-30 min) the ER Ca(2+) ATPase SERCA2b in monocytic cells (IC(50)=1.88 microM; p<0.05), thereby disrupting short-term Ca(2+) homeostasis (resting [Ca(2+)](cyto)=121.2+/-2.9% basal within 1h; p<0.05). However, extended Rosiglitazone treatment (72 h) induced dose-dependent SERCA2b up-regulation, and restored calcium homeostasis, in monocytic cells (SERCA2b mRNA: 138.7+/-5.7% basal (1 microM)/215.0+/-30.9% basal (10 microM); resting [Ca(2+)](cyto)=97.3+/-8.3% basal (10 microM)). As unfavourable cardiovascular outcomes, possibly related to disrupted cellular Ca(2+) homeostasis, have been linked to Rosiglitazone, this effect may be of clinical interest. In contrast, in PPRE-luciferase reporter-gene assays, Rosiglitazone induced non-dose-dependent PPARgamma-dependent effects (1 microM: 152.5+/-4.9% basal; 10 microM: 136.1+/-5.1% basal (p<0.05 for 1 microM vs. 10 microM)). Thus, we conclude that Rosiglitazone can exert PPARgamma-independent non-genomic effects, such as the SERCA2b inhibition seen here, but that long-term Rosiglitazone treatment did not perturb resting [Ca](cyto) in this study.

The tumour-suppressor Scribble dictates cell polarity during directed epithelial migration: regulation of Rho GTPase recruitment to the leading edge.

Altered expression of human Scribble is associated with invasive epithelial cancers, however, its role in tumour development remains unclear. Mutations in Drosophila Scribble result in loss of polarity, overproliferation and 3D-tumourous overgrowth of epithelial cells. Using complementation studies in Drosophila we recently demonstrated that expression of human Scribble can also regulate polarity and restrict tissue overgrowth. Here, we have undertaken a detailed study of human Scribble function in the polarized mammary cell line, MCF10A. We show that although Scribble does not seem to be required for apical-basal polarity or proliferation control in MCF10A cells, Scribble is essential for the control of polarity associated with directed epithelial cell migration. Scribble-depleted MCF10A cells show defective in vitro wound closure and chemotactic movement. The cells at the wound edge fail to polarize, show reduced lamellipodia formation and impaired recruitment of Cdc42 and Rac1 to the leading edge. Furthermore, we show that this function is relevant in vivo as Scribble mutant mice show defective epidermal wound healing. This data identifies an essential role for mammalian Scribble in the regulation of the polarity specifically involved in directed epithelial migration.

This is more difficult than we thought! The responsibility of scientists, managers and stakeholders to mitigate the unsustainability of marine fisheries.

The management of marine fisheries needs to undergo dramatic change in the new millennium, in response to the well-documented evidence of global overfishing and the general depletion of commercial fish stocks. The axioms of sustainable development and equilibrium productivity of wild ecosystems are identified as misleading concepts, which nonetheless underlie current approaches to the management of living marine resources. Current trends in marine fisheries landings worldwide provide little evidence of sustainability of marine resources under current management paradigms, where biological, economic and social aspects of fisheries are usually treated as different disciplines. While open-access conditions are less widespread than formerly, except for many straddling and highly migratory resources, fishers usually have access to the resource year-round throughout its range. Despite quotas, the nominal control of capacity and technical measures protecting juveniles, top-down management has generally been unable to prevent stock depletion, particularly of the older spawners that for demersal stocks often support recruitment. An integrated solution to the complexity of managing wild resources seems not to have been achieved. Any new paradigm should assert the basic unpredictability of fisheries at the system level and require a broader range of performance indicators to be incorporated into the decisional framework. This must reflect the non-equilibrium nature of marine systems, and give greater importance to resource (as opposed to harvest) continuity in the face of regime shifts, and promote habitat restoration and conservation of genetic resources. The new management framework requires co-management and collective decision-making to be incorporated within a precautionary and pre-negotiated management framework. This must explicitly recognize that decision-making occurs in conditions of model-based uncertainty and precautionary approaches should be incorporated at all levels, not least of which is to avoid the assumption that all resources can be harvested in a sustainable fashion through time. Redundancy in data inputs to management are needed to avoid the surprises that model-based sampling occasionally leads to, for example, when regime changes reduce productivity in response to climatic fluctuations. Emergency frameworks imposing non-discretionary rules must be invoked when overfishing and/or regime change trigger reference points indicating stock depletion. Non-discretionary recovery plans should then override rights-based systems and persist until fish populations recover to pre-established healthy levels, which may in turn need to await the return of a favourable regime. In fact, some stocks may require periodic rebuilding after regime-induced collapses or because of a combination of ecological or economic impacts, hence a constant harvest policy may not always be possible. It will probably also be necessary to discard the axiom that a stock should be available to harvesting throughout its range and seasonal cycle. Technological advances mean that time- and area-specific access rights are now practical options, through satellite monitoring of vessel operations, even offshore. More fundamentally, the basic axiom of "enlightened self interest" underlying current methods of management will need to be tempered by an increased ethical concern for the fragility of natural ecosystems.

Thermoregulation, plasma catecholamine and metabolite levels during submaximal work in individuals susceptible to malignant hyperpyrexia.

Measurements of body temperature, respiratory gas exchange, sweat evaporation rate and circulating levels of catecholamines, lactate, pyruvate, free fatty acids and glucose were made in seven patients susceptible to malignant hyperpyrexia (MHS) and in seven control subjects during 2 h of treadmill walking at 40% of maximum oxygen consumption. These studies took place in an ambient temperature of 22 degrees C. The MHS patients displayed the same thermoregulatory, plasma catecholamine and metabolic responses as the control subjects. The results of the present study suggest that non-competitive, low-intensity, steady-state exercise need not be contraindicated for MHS patients.