Prion aggregate structure in yeast cells is determined by the Hsp104-Hsp110 disaggregase machinery

Prions consist of misfolded proteins that have adopted an infectious amyloid conformation. In vivo, prion biogenesis is intimately associated with the protein quality control machinery. Using electron tomography, we probed the effects of the heat shock protein Hsp70 chaperone system on the structure of a model yeast [PSI+] prion in situ. Individual Hsp70 deletions shift the balance between fibril assembly and disassembly, resulting in a variable shell of nonfibrillar, but still immobile, aggregates at the surface of the [PSI+] prion deposits. Both Hsp104 (an Hsp100 disaggregase) and Sse1 (the major yeast form of Hsp110) were localized to this surface shell of [PSI+] deposits in the deletion mutants. Elevation of Hsp104 expression promoted the appearance of this novel, nonfibrillar form of the prion aggregate. Moreover, Sse1 was found to regulate prion fibril length. Our studies reveal a key role for Sse1 (Hsp110), in cooperation with Hsp104, in regulating the length and assembly state of [PSI+] prion fibrils in vivo

Anisamide-targeted cyclodextrin nanoparticles for siRNA delivery to prostate tumours in mice

A hepta-guanidino-β-cyclodextrin (G-CD), its hepta-PEG conjugate (G-CD-PEG), and the corresponding anisamide-terminated PEG conjugate (G-CD-PEG-AA) have been synthesised and compared as delivery vectors for siRNA to prostate cancer cells and tumours in vivo. The G-CD-PEG-AA.siRNA formulations (in which anisamide targets the sigma receptor), but not the non-targeted formulations, induced prostate cell-specific internalisation of siRNA resulting in approximately 80% knockdown in vitro of the reporter gene, luciferase. Following intravenous administration of the anisamide-targeted formulation in a mouse prostate tumour model significant tumour inactivation with corresponding reductions in the level of vascular endothelial growth factor (VEGF) mRNA were achieved, without demonstrating enhanced toxicity. This data imply significant potential for anisamide-conjugated cyclodextrin vectors for targeted delivery of therapeutic siRNAs in the treatment of prostate cancer

Femtosecond pulse generation in passively mode locked InAs quantum dot lasers

Optical pulse durations of an InAs two-section passively mode-locked quantum dot laser with a proton bombarded absorber section reduce from 8.4 ps at 250K to 290 fs at 20 K, a factor of 29, with a corresponding increase in optical bandwidth. Rate equation analysis of gain and emission spectra using rate equations suggests this is due to the very low emission rate of carriers to the wetting layer in the low temperature, random population regime which enables dots across the whole inhomogeneous distribution to act as independent oscillators. (C) 2013 AIP Publishing LLC

Intersecting brane world models from D8-branes on (T^2 x T^4/Z_3)/Omega R_1 type IIA orientifolds

We present orientifold models of type IIA string theory with D8-branes compactified on a two torus times a four dimensional orbifold. The orientifold group is chosen such that one coordinate of the two torus is reversed when applying worldsheet parity. RR tadpole cancellation requires D8-branes which wrap 1-cycles on the two torus and transform non-trivially under the orbifold group. These models are T-dual to orientifolds with D4-branes only which admit large volume compactifications. The intersections of the D8-branes are chosen in such a way that supersymmetry is broken in the open string sector and chiral fermions arise. Stability of the models is discussed in the context of NSNS tadpoles. Two examples with the SM gauge group and two left-right symmetric models are given.Comment: 30 pages, 7 figure

A click chemistry route to 2-functionalised PEGylated and cationic beta-cyclodextrins: co-formulation opportunities for siRNA delivery

A new approach to the synthesis of amphiphilic beta-cyclodextrins has used 'click' chemistry to selectively modify the secondary 2-hydroxyl group. The resulting extended polar groups can be either polycationic or neutral PEGylated groups and these two amphiphile classes are compatible in dual cyclodextrin formulations for delivery of siRNA. When used alone with an siRNA, a cationic cyclodextrin was shown to have good transfection properties in cell culture. Co-formulation with a PEGylated cyclodextrin altered the physicochemical properties of nanoparticles formed with siRNA. Improved particle properties included lower surface charges and reduced tendency to aggregate. However, as expected, the transfection efficiency of the cationic vector was lowered by co-formulation with the PEGylated cyclodextrin, requiring future surface modification of particles with targeting ligands for effective siRNA delivery

Cationic and PEGylated amphiphilic cyclodextrins: co-formulation opportunities for neuronal siRNA delivery

Optimising non-viral vectors for neuronal siRNA delivery presents a significant challenge. Here, we investigate a co-formulation, consisting of two amphiphilic cyclodextrins (CDs), one cationic and the other PEGylated, which were blended together for siRNA delivery to a neuronal cell culture model. Co-formulated CD-siRNA complexes were characterised in terms of size, charge and morphology. Stability in salt and serum was also examined. Uptake was determined by flow cytometry and toxicity was measured by MTT assay. Knockdown of a luciferase reporter gene was used as a measure of gene silencing efficiency. Incorporation of a PEGylated CD in the formulation had significant effects on the physical and biological properties of CD. siRNA complexes. Co-formulated complexes exhibited a lower surface charge and greater stability in a high salt environment. However, the inclusion of the PEGylated CD also dramatically reduced gene silencing efficiency due to its effects on neuronal uptake. The co-formulation strategy for cationic and PEGylated CDs improved the stability of the CD. siRNA delivery systems, although knockdown efficiency was impaired. Future work will focus on the addition of targeting ligands to the co-formulated complexes to restore transfection capabilities

Click-modified cyclodextrins as non-viral vectors for neuronal siRNA delivery

RNA interference (RNAi) holds great promise as a strategy to further our understanding of gene function in the central nervous system (CNS) and as a therapeutic approach for neurological and neurodegenerative diseases. However, the potential for its use is hampered by the lack of siRNA delivery vectors, which are both safe and highly efficient. Cyclodextrins have been shown to be efficient and low toxicity gene delivery vectors in various cell types in vitro. However, to date they have not been exploited for delivery of oligonucleotides to neurons. To this end, a modified β-cyclodextrin (CD) vector was synthesised, which complexed siRNA to form cationic nanoparticles of less than 200nm in size. Furthermore, it conferred stability in serum to the siRNA cargo. The in vitro performance of the CD in both immortalised hypothalamic neurons and primary hippocampal neurons was evaluated. The CD facilitated high levels of intracellular delivery of labelled siRNA, whilst maintaining at least 80% cell viability. Significant gene knockdown was achieved, with a reduction in luciferase expression of up to 68% and a reduction in endogenous glyceraldehyde phosphate dehydrogenase (GAPDH) expression of up to 40%. To our knowledge, this is the first time that a modified CD has been used as a safe and efficacious vector for siRNA delivery into neuronal cells

Body mass index mortality paradox in chronic kidney disease patients with suspected cardiac chest pain

Background: Chronic kidney disease (CKD) is a silent clinical condition associated with adverse comorbidity and high cardiovascular disease (CVD) risk. An inverse relationship with body mass index (BMI) and mortality has been demonstrated in hemodialysis patients. However, it is unclear if this risk‐factor paradox is evident in non‐dialysis CKD patients. The aims of this study were to explore the relationship between, nutritional status, markers of inflammation, autonomic and cardiac function with BMI. Longitudinal follow‐up explored the relationship between BMI and allcause mortality. Methods: 211‐consecutive CKD patients referred for dobutamine stress echocardiography to detect or exclude myocardial ischemia were recruited. BMI, albumin, C‐reactive protein (CRP) and haemoglobin (Hb) were recorded as markers of nutritional and inflammatory status. Left ventricular ejection fraction (LVEF) and heart rate variability (HRV) as an indicator of cardiac function was recorded. All subjects were followed prospectively until November 2014 and study end‐point was all‐cause mortality. Results: BMI was inversely associated with CKD status. After covariate adjustment, this association remained. During a mean follow‐up period of 3.3±0.9 years there were 35 deaths (17%). BMI was inversely associated with all‐cause mortality (HR 0.81, 95% CI 0.71‐0.9). Other important independent predictors of mortality were heart rate variability (HR 0.98, 95% CI 0.97‐0.99), myocardial ischemia (HR 1.37, 95% CI 1.17‐1.81), and albumin (HR 0.86, 95% CI 0.81‐ 0.92). Conclusions: The presence of a BMI paradox exists in non‐dialysis CKD patients. This risk‐factor paradox was an independent predictor of all‐cause mortality and may have significant clinical implications relevant to screening, assessment and treatment and requires further study