A Study on the Analytical Sensitivity of 6 BSE Tests Used by the Canadian BSE Reference Laboratory

Bovine spongiform encephalopathy (BSE) surveillance programs have been employed in numerous countries to monitor BSE prevalence and to protect animal and human health. Since 1999, the European Commission (EC) authorized the evaluation and approval of 20 molecular based tests for the rapid detection of the pathological prion protein (PrPsc) in BSE infection. The diagnostic sensitivity, convenience, and speed of these tests have made molecular diagnostics the preferred method for BSE surveillance. The aim of this study was to determine the analytical sensitivity of 4 commercially available BSE rapid-test kits, including the Prionics®-Check WESTERN, the Prionics® Check-PrioSTRIP™, the BioRad® TeSeE™ ELISA, and the IDEXX® HerdChek™ EIA. Performances of these tests were then compared to 2 confirmatory tests, including the BioRad® TeSeE™ Western Blot and the modified Scrapie Associated Fibrils (SAF)/OIE Immunoblot. One 50% w/v homogenate was made from experimentally generated C-type BSE brain tissues in ddH2O. Homogenates were diluted through a background of BSE-negative brainstem homogenate. Masses of both positive and negative tissues in each dilution were calculated to maintain the appropriate tissue amounts for each test platform. Specific concentrated homogenization buffer was added accordingly to maintain the correct buffer condition for each test. ELISA-based tests were evaluated using their respective software/detection platforms. Blot-protocols were evaluated by manual measurements of blot signal density. Detection limitations were determined by fitted curves intersecting the manufacturers' positive/negative criteria. The confirmatory SAF Immunoblot displayed the highest analytical sensitivity, followed by the IDEXX® HerdChek™ EIA, Bio-Rad® TeSeE™ Western Blot, the Bio-Rad® TeSeE™ ELISA, Prionics®-Check PrioSTRIP™, and Prionics®-Check WESTERN™, respectively. Although the tests performed at different levels of sensitivity, the most sensitive and least sensitive of the rapid tests were separated by 2 logs in analytical sensitivity, meeting European performance requirements. All rapid tests appear suitable for targeted BSE surveillance programs, as implemented in Canada

Synchronous vs Asynchronous Chain Motion in α-Synuclein Contact Dynamics

α-Synuclein (α-syn) is an intrinsically unstructured 140-residue neuronal protein of uncertain function that is implicated in the etiology of Parkinson’s disease. Tertiary contact formation rate constants in α-syn, determined from diffusion-limited electron-transfer kinetics measurements, are poorly approximated by simple random polymer theory. One source of the discrepancy between theory and experiment may be that interior-loop formation rates are not well approximated by end-to-end contact dynamics models. We have addressed this issue with Monte Carlo simulations to model asynchronous and synchronous motion of contacting sites in a random polymer. These simulations suggest that a dynamical drag effect may slow interior-loop formation rates by about a factor of 2 in comparison to end-to-end loops of comparable size. The additional deviations from random coil behavior in α-syn likely arise from clustering of hydrophobic residues in the disordered polypeptide

Evidence of Strong-Coupled Superconductivity in CaC6 from Tunneling Spectroscopy

Point-contact tunneling on CaC$_6$ crystals reproducibly reveals superconducting gaps, $\Delta$, of 2.3$\pm$0.2 meV which are $\sim$~40% larger than earlier reports. That puts CaC$_6$ into the class of very strong-coupled superconductors since 2$\Delta$/kT$_c\sim$~4.6. Thus soft Ca phonons will be primarily involved in the superconductivity, a conclusion that explains the large Ca isotope effect found recently for CaC$_6$. Consistency among superconductor-insulator-normal metal (SIN), SIS and Andreev reflection (SN) junctions reinforces the intrinsic nature of this result.Comment: 2nd version, 4 pages, 4 figures, re-submitted to Physical Review Letter

Effect of stress state and simultaneous hot corrosion on the crack propagation and fatigue life of single crystal superalloy CMSX-4

Operating conditions within industrial gas turbines are changing in response to pressures to reduce environmental impact and enable use of renewable sources. This is driving an increase in the operational temperatures and pressures of combustion in turbine systems. Additionally, diverse operating environments can result in higher sulphur and trace metal contaminant levels, exacerbating hot corrosion in GT systems. Low cycle fatigue (LCF) cycling can also be intensified as a result of increased start/stop shutdowns. The combined effects of hot corrosion and stress are experimentally studied on CMSX-4 single crystal (SC) γ/γ' system under both fatigue and static stress conditions, with either a multi-axial bending or uniaxial stress state. The associated stress intensity thresholds (KTH) under the various stress conditions were evaluated using finite element analysis (FEA). Cracking was observed both under static and fatigue stress conditions in a hot corrosion environment. Crack morphologies were analysed using SEM techniques. Bending stresses and fatigue cycles demonstrated increased crack propagation in the presence of hot corrosion with static uniaxial stresses showing the longest nucleation times and lowest propagation rates

Self-heating effect in intrinsic tunneling spectroscopy of HgBr2 intercalated Bi2.1Sr1.4Ca1.5Cu 2O8+δ single crystals

We report tunneling results in intrinsic Josephson junction (IJJ) stacks fabricated in the form of square micromesas on HgBr2 intercalated Bi2.1Sr1.4Ca1.5Cu2O 8+δ (Bi2212) single crystals using photolithography and Ar ion milling techniques. Self-heating is the most common problem encountered in interlayer tunneling and it is likely to reduce the reliability of IJJ data. Although intercalation reduces heating a hundredfold, it still needs to be minimized substantially in order to approach the authentic superconducting energy gap observed by tunneling using more conventional junctions. We report tunneling characteristics of two mesas with the same height but different sizes (5 × 5 μm2 and 10 × 10 μm2) to show that heating effects are strongly related to IJJ stack size. For the smaller mesa, we observed an energy gap close to that seen in single SIN (S: superconductor, I: insulator, N: normal metal) and SIS break junctions as well as the dip and hump structures at high bias. The subgap data of 5 × 5 μm2 mesa were successfully fit with a momentum averaged d-wave model using convenient parameters. Thus our data is consistent with the predominant pairing symmetry suggested by point contact tunneling, break junction, scanning tunneling microscopy/spectroscopy and angle resolved photoemission measurements in Bi2Sr2CaCu2O8+δ

Geometrical analysis of cytochrome c unfolding

A geometrical model has been developed to study the unfolding of iso-1 cytochrome c. The model draws on the crystallographic data reported for this protein. These data were used to calculate the distance between specific residues in the folded state, and in a sequence of extended states defined by n = 3, 5, 7, 9, 11, 13, and 15 residue units. Exact calculations carried out for each of the 103 residues in the polypeptide chain demonstrate that different regions of the chain have different unfolding histories. Regions where there is a persistence of compact structures can be identified, and this geometrical characterization is fully consistent with analyses of time-resolved fluorescence energy-transfer (TrFET) data using dansyl-derivatized cysteine side-chain probes at positions 39, 50, 66, 85, and 99. The calculations were carried out assuming that different regions of the polypeptide chain unfold synchronously. To test this assumption, lattice Monte Carlo simulations were performed to study systematically the possible importance of asynchronicity. Calculations show that small departures from synchronous dynamics can arise if displacements of residues in the main body of the chain are much more sluggish than near-terminal residues

Counterintuitive Consequence of Heating in Strongly-Driven Intrinsic Junctions of Bi₂Sr₂CaCu₂O\u3csub\u3e8+δ\u3c/sub\u3e mesas

Anomalously high and sharp peaks in the conductance of intrinsic Josephson junctions in Bi2 Sr2 CaCu2 O 8+δ (Bi2212) mesas have been commonly interpreted as superconducting energy gaps but here we show they are a result of strong self-heating. This conclusion follows directly from a comparison to the equilibrium gap measured by tunneling in single break junctions on equivalent crystals. As the number of junctions in the mesa, N, and thus heating increase, the peak voltages decrease and the peak width abruptly sharpens for N≥12. Clearly these widely variable features vs N cannot all represent the equilibrium properties. Our data imply that the sharp peaks represent a transition to the normal state. That it occurs at the same dissipated power for N=12-30 strongly implicates heating as the cause. Although peak sharpening due to heating is counterintuitive, as tunneling spectra usually broaden at higher temperatures, a lateral temperature gradient, leading to coexistence of normal hot spots and superconductive regions, qualitatively explains the behavior. However, a more uniform temperature profile cannot be ruled out. As the peak\u27s width and voltage in our shortest mesa (N=6) are more consistent with the break junction data, we propose a figure of merit for Bi2212 mesas, the relative conductance peak width, such that small values signal a crossover into the strong self-heating regime

The fundamental cycle of concept construction underlying various theoretical frameworks

In this paper, the development of mathematical concepts over time is considered. Particular reference is given to the shifting of attention from step-by-step procedures that are performed in time, to symbolism that can be manipulated as mental entities on paper and in the mind. The development is analysed using different theoretical perspectives, including the SOLO model and various theories of concept construction to reveal a fundamental cycle underlying the building of concepts that features widely in different ways of thinking that occurs throughout mathematical learning

Reply to Comment on \u27Counterintuitive Consequence of Heating in Strongly-Driven Intrinsic Junctions of Bi₂Sr₂CaCu₂O \u3csub\u3e8+δ\u3c/sub\u3e Mesas\u27

The main criticism raised in the preceding Comment concerns our suggestion that sharp conduction peaks in Bi2Sr2CaCu2O 8+δ mesas, along with absent dip-hump features, may, in general, be a result of self-heating. The author points to the variety of experimental configurations, matrix-element effects, and doping dependencies that might allow a diversity of conductance spectra. We argue that numerous mesa studies (with fixed matrix elements) firmly establish the systematic development of sharp conductance peaks with increased self-heating, and thus, the issue of nonuniversality of tunneling characteristics is not relevant. The author mentions a number of studies that show that the mesa is superconducting near the conductance peak voltage. This is not in dispute and indicates a misinterpretation of our analysis that is clarified here. To address further comments on the technical details of our heating model, we reiterate that our conclusions are independent of our model but rather are based solely on experimental data that are not in dispute