MAG induces regulated intramembrane proteolysis of the p75 neurotrophin receptor to inhibit neurite outgrowth.

The three known inhibitors of axonal regeneration present in myelin--MAG, Nogo, and OMgp--all interact with the same receptor complex to effect inhibition via protein kinase C (PKC)-dependent activation of the small GTPase Rho. The transducing component of this receptor complex is the p75 neurotrophin receptor. Here we show that MAG binding to cerebellar neurons induces alpha- and then gamma-secretase proteolytic cleavage of p75, in a protein kinase C-dependent manner, and that this cleavage is necessary for both activation of Rho and inhibition of neurite outgrowth.

Activated CREB is sufficient to overcome inhibitors in myelin and promote spinal axon regeneration in vivo.

Inhibitors in myelin play a major role in preventing spontaneous axonal regeneration after CNS injury. Elevation of cAMP overcomes this inhibition, in a transcription-dependent manner, through the upregulation of Arginase I (Arg I) and increased synthesis of polyamines. Here, we show that the cAMP effect requires activation of the transcription factor cAMP response element binding protein (CREB) to overcome myelin inhibitors; a dominant-negative CREB abolishes the effect, and neurons expressing a constitutively active form of CREB are not inhibited. Activation of CREB is also required for cAMP to upregulate Arg I, and the ability of constitutively active CREB to overcome inhibition is blocked by an inhibitor of polyamine synthesis. Finally, expression of constitutively active CREB in DRG neurons is sufficient to promote regeneration of subsequently lesioned dorsal column axons. These results indicate that CREB plays a central role in overcoming myelin inhibitors and so encourages regeneration in vivo.

The cytokine interleukin-6 is sufficient but not necessary to mimic the peripheral conditioning lesion effect on axonal growth.

Lesioning the peripheral branch of a dorsal root ganglion (DRG) neuron before injury of the central branch of the same neuron enables spontaneous regeneration of these spinal axons. This effect is cAMP and transcription dependent. Here, we show that the cytokine interleukin-6 (IL-6) is upregulated in DRG neurons after either a conditioning lesion or treatment with dibutyryl-cAMP. In culture, IL-6 allows neurons to grow in the presence of inhibitors of regeneration present in myelin. Importantly, intrathecal delivery of IL-6 to DRG neurons blocks inhibition by myelin both in vitro and in vivo, effectively mimicking the conditioning lesion. Blocking IL-6 signaling has no effect on the ability of cAMP to overcome myelin inhibitors. Consistent with this, IL-6-deficient mice respond to a conditioning lesion as effectively as wild-type mice. We conclude that IL-6 can mimic both the cAMP effect and the conditioning lesion effect but is not an essential component of either response.

Endothelial repair in stented arteries is accelerated by inhibition of Rho-associated protein kinase.

AIMS: Stent deployment causes endothelial cells (EC) denudation, which promotes in-stent restenosis and thrombosis. Thus endothelial regrowth in stented arteries is an important therapeutic goal. Stent struts modify local hemodynamics, however the effects of flow perturbation on EC injury and repair are incompletely understood. By studying the effects of stent struts on flow and EC migration, we identified an intervention that promotes endothelial repair in stented arteries. METHODS AND RESULTS: In vitro and in vivo models were developed to monitor endothelialization under flow and the influence of stent struts. A 2D parallel-plate flow chamber with 100 µm ridges arranged perpendicular to the flow was used. Live cell imaging coupled to computational fluid dynamic simulations revealed that EC migrate in the direction of flow upstream from the ridges but subsequently accumulate downstream from ridges at sites of bidirectional flow. The mechanism of EC trapping by bidirectional flow involved reduced migratory polarity associated with altered actin dynamics. Inhibition of Rho-associated protein kinase (ROCK) enhanced endothelialization of ridged surfaces by promoting migratory polarity under bidirectional flow (P < 0.01). To more closely mimic the in vivo situation, we cultured EC on the inner surface of polydimethylsiloxane tubing containing Coroflex Blue stents (65 µm struts) and monitored migration. ROCK inhibition significantly enhanced EC accumulation downstream from struts under flow (P < 0.05). We investigated the effects of ROCK inhibition on re-endothelialization in vivo using a porcine model of EC denudation and stent placement. En face staining and confocal microscopy revealed that inhibition of ROCK using fasudil (30 mg/day via osmotic minipump) significantly increased re-endothelialization of stented carotid arteries (P < 0.05). CONCLUSIONS: Stent struts delay endothelial repair by generating localized bidirectional flow which traps migrating EC. ROCK inhibitors accelerate endothelial repair of stented arteries by enhancing EC polarity and migration through regions of bidirectional flow.

Heart rate reduction with ivabradine promotes shear stress-dependent anti-inflammatory mechanisms in arteries.

Blood flow generates wall shear stress (WSS) which alters endothelial cell (EC) function. Low WSS promotes vascular inflammation and atherosclerosis whereas high uniform WSS is protective. Ivabradine decreases heart rate leading to altered haemodynamics. Besides its cardio-protective effects, ivabradine protects arteries from inflammation and atherosclerosis via unknown mechanisms. We hypothesised that ivabradine protects arteries by increasing WSS to reduce vascular inflammation. Hypercholesterolaemic mice were treated with ivabradine for seven weeks in drinking water or remained untreated as a control. En face immunostaining demonstrated that treatment with ivabradine reduced the expression of pro-inflammatory VCAM-1 (p<0.01) and enhanced the expression of anti-inflammatory eNOS (p<0.01) at the inner curvature of the aorta. We concluded that ivabradine alters EC physiology indirectly via modulation of flow because treatment with ivabradine had no effect in ligated carotid arteries in vivo, and did not influence the basal or TNFα-induced expression of inflammatory (VCAM-1, MCP-1) or protective (eNOS, HMOX1, KLF2, KLF4) genes in cultured EC. We therefore considered whether ivabradine can alter WSS which is a regulator of EC inflammatory activation. Computational fluid dynamics demonstrated that ivabradine treatment reduced heart rate by 20 % and enhanced WSS in the aorta. In conclusion, ivabradine treatment altered haemodynamics in the murine aorta by increasing the magnitude of shear stress. This was accompanied by induction of eNOS and suppression of VCAM-1, whereas ivabradine did not alter EC that could not respond to flow. Thus ivabradine protects arteries by altering local mechanical conditions to trigger an anti-inflammatory response.

Modelling the glycocalyx-endothelium-erythrocyte interaction in the microcirculation: a computational study.

A novel, coarse-grained, single-framework 'Eulerian' model for blood flow in the microvascular circulation is presented and used to estimate the variations in flow properties that accrue from all of the following: (i) wall position variation, associated with the endothelial cells' (ECs) shape, (ii) glycocalyx layer (GL) effects and (iii) the particulate nature of blood. We stress that our new model is fully coupled and uses only a single Eulerian computational framework to recover complex effects, dispensing altogether with the need for, e.g. re-meshing and advected sets of Lagrangian points. Physically, blood is modelled as a two-component, incompressible fluid - the plasma and corpuscular elements dispersed in it. The latter are modelled as deformable liquid droplets of increased viscosity. Interfacial membrane effects are present to mimic key blood properties and to avoid droplets' coalescence. The model is encapsulated within a multi-component lattice Boltzmann method that uses a sub-lattice 'wavy wall' closure to represent the ECs. Between this boundary and the flow domain, the model incorporates a coarse-grained representation of the endothelial GL, which is known to cover microvessel walls. The endothelial glycocalyx is modelled as a medium of variable and adaptive porosity, with approaching droplets being subject to a repulsive elastic force. Numerical simulations are presented to show the combined and simultaneous influence on fundamental flow properties of the EC wall undulation, the glycocalyx compression and repulsion and the particulate nature of blood. Several characteristic hemodynamical features of microvessel flow are successfully reproduced, including the deformability of particulates and the Fahraeus-Lindqvist effect. Moreover, the importance of modelling the GL is manifest in the magnitude of and the temporal variations in the flow rate and wall shear stresses.

Central venous catheter insertion by a clinical nurse consultant or anaesthetic medical staff: a single-centre observational study.

OBJECTIVE: To compare clinical outcomes of elective central venous catheter (CVC) insertions performed by either a clinical nurse consultant (CNC) or anaesthetic medical staff (AMS). DESIGN, SETTING AND PARTICIPANTS: Prospective audit of a convenience sample of consecutive CVC insertions between July 2005 and October 2007 at a metropolitan teaching hospital in Sydney, Australia. The sample included all outpatients and inpatients requiring a CVC for either acute or chronic conditions. MAIN OUTCOME MEASURES: Number of CVC lines inserted; differences between outcomes in the CNC and AMS groups; complications during and after insertion. RESULTS: Over a 28-month period, 245 CVCs were inserted by AMS and 123 by the CNC. The most common indications for CVC placement in both groups were for the treatment of oncology and autoimmune disorders (61%) and for antibiotic therapy (27%). Other indications were parenteral nutrition (2%) and other therapies (10%). There was no significant difference in complications on insertion between the CNC and AMS groups. AMS failed to obtain access in 12 attempted procedures compared with eight by the CNC. The rate of CVCs investigated for infection was twice as high in the AMS group as in the CNC group (19% v 8%). The confirmed catheter-related bloodstream infection (CRBSI) rate was 2.5/1000 catheters in the AMS group and 0.4/1000 catheters in the CNC group (P = 0.04). CONCLUSION: Insertion outcomes were favourable in both the AMS and CNC groups. Infection outcomes differed between groups, with a higher rate of CRBSI in the AMS group.

A role for cAMP in regeneration of the adult mammalian CNS.

Injury to the adult mammalian central nervous system (CNS) often results in permanent loss of sensory and motor function. This is due to the failure of injured axons to regenerate. The inhibitory nature of the CNS can be attributed to several factors, including formation of the glial scar, the presence of several molecules, associated with myelin, which inhibit axonal regrowth, and the intrinsic growth state of these neurons. Encouraging regeneration in the adult mammalian CNS therefore will require targeting one or all of these factors following injury. Here we illustrate recent work from our laboratory that identifies some of the signalling components involved in modulation of the intrinsic growth state of adult neurons. When activated, these signalling pathways can induce axonal regeneration in the presence of the myelin-associated inhibitors both in vitro and in vivo.

Endometrial adenocarcinoma following the insertion of a Mirena IUCD.

BACKGROUND: The case histories of two patients who developed endometrial adenocarcinoma with a Mirena intrauterine system (IUS) in place are reported. CASE: Two patients had a Mirena IUS inserted to treat abnormal vaginal bleeding. After 12 and 36 months with the device in place they presented with a recurrence of their heavy, irregular menstrual bleeding. Endometrial biopsies confirmed the presence of endometrial adenocarcinoma, and hysterectomies were performed. In one patient the cancer was localized and below the stem of the device, in the other metastatic disease had developed. CONCLUSION: These cases raises important issues regarding the use of the Mirena IUS to treat menorrhagia.