ANNINE-6plus, a voltage-sensitive dye with good solubility, strong membrane binding and high sensitivity

We present a novel voltage-sensitive hemicyanine dye ANNINE-6plus and describe its synthesis, its properties and its voltage-sensitivity in neurons. The dye ANNINE-6plus is a salt with a double positively charged chromophore and two bromide counterions. It is derived from the zwitterionic dye ANNINE-6. While ANNINE-6 is insoluble in water, ANNINE-6plus exhibits a high solubility of around 1 mM. Nonetheless, it displays a strong binding to lipid membranes. In contrast to ANNINE-6, the novel dye can be used to stain cells from aqueous solution without surfactants or organic solvents. In neuronal membranes, ANNINE-6plus exhibits the same molecular Stark effect as ANNINE-6. As a consequence, a high voltage-sensitivity is achieved with illumination and detection in the red end of the excitation and emission spectra, respectively. ANNINE-6plus will be particularly useful for sensitive optical recording of neuronal excitation when organic solvents and surfactants must be avoided as with intracellular or extracellular staining of brain tissue

New Role, New Country: introducing US physician assistants to Scotland

This paper draws from research commissioned by the Scottish Executive Health Department (SEHD). It provides a case study in the introduction of a new health care worker role into an already well established and "mature" workforce configuration It assesses the role of US style physician assistants (PAs), as a precursor to planned "piloting" of the PA role within the National Health Service (NHS) in Scotland

Dual Triumphalist Heritage Narrative And The Sungai Buloh Leprosy Settlement

Unlike other heritage movements in Malaysia, which are largely ethnicbased and culture obsessed (Cartier 1996; Worden 2001), the preservation movement of the Sungai Bulah Leprosy Settlement (SBLS thereafter), also widely referred to as the "Valley of Hope", 1 is concerned with the conservation of a site that is associated with a socially stigmatised disease. Built at a jungle fringe in Selangor in 1930, SBLS was constructed as a place for the treatment, and forced isolation from wider society, of people suffering from leprosy. Although leprosy knows no racial boundaries as people of any background can be afflicted with the disease, nearly eighty per cent of the patients admitted to SBLS have been ethnic Chinese. Of the rest, about fifteen per cent were ethnic Malays with ethnic Indians making up five per cent. Former patients who were cured but left with differing degrees of disfigurement and disability are also residents of the SBLS today.2 SBLS's population reached its peal<. with 2400 people in 1958, but today their number is just slightly over one hundred (JoshuaRaghavar 1983; Wong and Phang 2006)

Total Energy Expenditure and Body Composition Changes Following Peripheral Blood Stem Cell Transplantation and Participation in an Exercise Programme

The purpose of this investigation was to assess changes in total energy expenditure (TEE), body weight (BW) and body composition following a peripheral blood stem cell transplant and following participation in a 3-month duration, moderate-intensity, mixed-type exercise programme. The doubly labelled and singly labelled water methods were used to measure TEE and total body water (TBW). Body weight and TBW were then used to calculate percentage body fat (%BF), and fat and fat-free mass (FFM). TEE and body composition measures were assessed pretransplant (PI), immediately post-transplant (PII) and 3 months post-PII (PIII). Following PII, 12 patients were divided equally into a control group (CG) or exercise intervention group (EG). While there was no change in TEE between pre- and post-transplant, BW (P<0.01) and FFM (P<0.05) significantly decreased during the same period. Participation in the exercise programme led to increases in TEE to levels that were both higher than pre- and post-transplant measures (P<0.01). By PIII, the exercising patients also showed gains in FFM (P<0.01) in association with a reduction in %BF (P<0.05). Exercise has a functionally important role in preserving and increasing skeletal mass in the rehabilitation phase of cancer patients

Incomplete pneumolysin oligomers form membrane pores

Pneumolysin is a member of the cholesterol-dependent cytolysin (CDC) family of pore-forming proteins that are produced as water-soluble monomers or dimers, bind to target membranes and oligomerize into large ring-shaped assemblies comprising approximately 40 subunits and approximately 30 nm across. This pre-pore assembly then refolds to punch a large hole in the lipid bilayer. However, in addition to forming large pores, pneumolysin and other CDCs form smaller lesions characterized by low electrical conductance. Owing to the observation of arc-like (rather than full-ring) oligomers by electron microscopy, it has been hypothesized that smaller oligomers explain smaller functional pores. To investigate whether this is the case, we performed cryo-electron tomography of pneumolysin oligomers on model lipid membranes. We then used sub-tomogram classification and averaging to determine representative membrane-bound low-resolution structures and identified pre-pores versus pores by the presence of membrane within the oligomeric curve. We found pre-pore and pore forms of both complete (ring) and incomplete (arc) oligomers and conclude that arc-shaped oligomeric assemblies of pneumolysin can form pores. As the CDCs are evolutionarily related to the membrane attack complex/perforin family of proteins, which also form variably sized pores, our findings are of relevance to that class of proteins as well