Microwave-Accelerated Metal-Enhanced Fluorescence (MAMEF): Application to Ultra Fast and Sensitive Clinical Assays

In this rapid communication we describe an exciting platform technology that promises to fundamentally address two underlying constraints of modern assays and immunoassays, namely sensitivity and rapidity. By combining the use of Metal-enhanced Fluorescence (MEF) with low power microwave heating (Mw), we can significantly increase the sensitivity of surface assays as well as >95% kinetically complete the assay within a few seconds. This technology is subsequently likely to find significant importance in certain clinical assays, such as in the clinical assessment of myoglobin, where both the assay rapidity and sensitivity are paramount for the assessment and treatment of acute myocardial infarction.     

Applications of cavity ring-down spectroscopy to high precision isotope ratio measurement of 13C/12C in carbon dioxide

Recent measurements of carbon isotopes in carbon dioxide using near-infrared, diode-laser-based cavity ring-down spectroscopy (CRDS) are presented. The CRDS system achieved good precision, often better than 0.2 per thousand, for 4% CO2 concentrations, and also achieved 0.15-0.25 per thousand precision in a 78 min measurement time with cryotrap-based pre-concentration of ambient CO2 concentrations (360 ppmv). These results were obtained with a CRDS system possessing a data rate of 40 ring-downs per second and a loss measurement of 4.0 x 10(-11) cm(-1) Hz(-1/2). Subsequently, the measurement time has been reduced to under 10 min. This standard of performance would enable a variety of high concentration (3-10%) isotopic measurements, such as medical human breath analysis or animal breath experiments. The extension of this ring-down to the 2 microm region would enable isotopic analysis at ambient concentrations, which, combined with the small size, robust design, and potential for frequent measurements at a remote site, make CRDS technology attractive for remote atmospheric measurement applications.     

Tri-partite complex for axonal transport drug delivery achieves pharmacological effect

Background

Targeted delivery of pharmaceutical agents into selected populations of CNS (Central Nervous System) neurons is an extremely compelling goal. Currently, systemic methods are generally used for delivery of pain medications, anti-virals for treatment of dermatomal infections, anti-spasmodics, and neuroprotectants. Systemic side effects or undesirable effects on parts of the CNS that are not involved in the pathology limit efficacy and limit clinical utility for many classes of pharmaceuticals. Axonal transport from the periphery offers a possible selective route, but there has been little progress towards design of agents that can accomplish targeted delivery via this intraneural route. To achieve this goal, we developed a tripartite molecular construction concept involving an axonal transport facilitator molecule, a polymer linker, and a large number of drug molecules conjugated to the linker, then sought to evaluate its neurobiology and pharmacological behavior.

Results

We developed chemical synthesis methodologies for assembling these tripartite complexes using a variety of axonal transport facilitators including nerve growth factor, wheat germ agglutinin, and synthetic facilitators derived from phage display work. Loading of up to 100 drug molecules per complex was achieved. Conjugation methods were used that allowed the drugs to be released in active form inside the cell body after transport. Intramuscular and intradermal injection proved effective for introducing pharmacologically effective doses into selected populations of CNS neurons. Pharmacological efficacy with gabapentin in a paw withdrawal latency model revealed a ten fold increase in half life and a 300 fold decrease in necessary dose relative to systemic administration for gabapentin when the drug was delivered by axonal transport using the tripartite vehicle.

Conclusion

Specific targeting of selected subpopulations of CNS neurons for drug delivery by axonal transport holds great promise. The data shown here provide a basic framework for the intraneural pharmacology of this tripartite complex. The pharmacologically efficacious drug delivery demonstrated here verify the fundamental feasibility of using axonal transport for targeted drug delivery.     

Structural Aspects of the O-glycosylation Linkage in Glycopeptides via MD Simulations and Comparison with NMR Experiments

A powerful conformational searching and enhanced sampling simulation method, and unbiased molecular dynamics simulations have been used along with NMR spectroscopic observables to provide a detailed structural view of O-glycosylation. For four model systems, the force-field parameters can accurately predict experimental NMR observables (J couplings and NOE's). This enables us to derive conclusions based on the generated ensembles, in which O-glycosylation affects the peptide backbone conformation by forcing it towards to an extended conformation. An exception is described for β-GalNAc-Thr where the α content is increased and stabilized via hydrogen bonding between the sugar and the peptide backbone, which was not observed in the rest of the studied systems. These observations might offer an explanation for the evolutionary preference of α-linked GalNAc glycosylation instead of a β link.     

Changes in Silk Feedstock Rheology during Cocoon Construction: The Role of Calcium and Potassium Ions

Variation in silk feedstocks is a barrier both to our understanding of natural spinning and biomimetic endeavors. To address this, compositional changes are investigated in feedstock specimens from the domesticated silkworm (Bombyx mori). It is found that the feedstock viscosity decreased systematically by over two orders of magnitude during cocoon construction. Potential factors such as protein concentration, molecular weight, pH, or the presence of trehalose are excluded, whereas a clear correlation appear between viscosity and the relative concentrations of Ca²⁺ and K⁺ ions. It is expected that Ca²⁺ ions would favor “salt bridges” between acidic (Asp and Glu) amino acids, leading to an increased viscosity, whereas K⁺ ions would compete for these sites, thereby reducing viscosity. Thus, these findings suggest a simple, systematic yet sophisticated control of feedstock viscosity in the silkworm, which in turn can be applied to future industrial silk production.     

Differential Scanning Calorimetry of Native Silk Feedstock

Native silk proteins, extracted directly from the silk gland prior to spinning, offer access to a naturally hydrated protein that has undergone little to no processing. Combined with differential scanning calorimetry (DSC), it is possible to probe the thermal stability and hydration status of silk and thus investigate its denaturation and solidification, echoing that of the natural spinning process. It is found that native silk is stable between ?10 °C and 55 °C, and both the high‐temperature enthalpy of denaturation (measured via modulated temperature DSC) and a newly reported low‐temperature ice‐melting transition may serve as useful quality indicators in the future for artificial silks. Finally, compared to albumin, silk's denaturation enthalpy is much lower than expected, which is interpreted within a recently proposed entropic desolvation framework which can serve to unveil the low‐energy aquamelt processing pathway.     

Immunoassay utilizing imaging surface plasmon resonance for the detection of cyclopiazonic acid (CPA) in maize and cheese

α-Cyclopiazonic acid (CPA) is a tremorgenic mycotoxin produced by Aspergillus and Penicillium fungal species, commonly found on agricultural commodities or fermented food products. A sensitive and rapid imaging surface plasmon resonance (iSPR) assay was developed to detect CPA in maize and cheese by combining an indirect competitive immunoassay and signal amplification based upon a secondary antibody (Ab₂) conjugated with gold nanoparticles. Matrix-matched calibration curves were used to determine CPA content in maize and cheese samples. Recoveries, at two spiking levels in maize and cheese, were 89 to 126%, with standard deviations of repeatability (RSDr) of less than 16%. The limits of detection were 17 and 6 μg/kg in maize and cheese, respectively. To separate the CPA-contaminated samples from uncontaminated samples, a cutoff validation level of 40 μg/kg was introduced. The assay was applied to samples of naturally contaminated maize and was compared with competitive inhibition enzyme-linked immunosorbent assay (CI-ELISA). This is the first report to detect CPA using an immuno-biosensor iSPR format.

     

A mechanism for mitigation of blade–vortex interaction using leading edge blowing flow control

The interaction of a vortical unsteady flow with structures is often encountered in engineering applications. Such flow structure interactions (FSI) can be responsible for generating significant loads and can have many detrimental structural and acoustic side effects, such as structural fatigue, radiated noise and even catastrophic results. Amongst the different types of FSI, the parallel blade–vortex interaction (BVI) is the most common, often encountered in helicopters and propulsors. In this work, we report on the implementation of leading edge blowing (LEB) active flow control for successfully minimizing the parallel BVI. Our results show reduction of the airfoil vibrations up to 38% based on the root-mean-square of the vibration velocity amplitude. This technique is based on displacing an incident vortex using a jet issued from the leading edge of a sharp airfoil effectively increasing the stand-off distance of the vortex from the body. The effectiveness of the method was experimentally analyzed using time-resolved digital particle image velocimetry (TRDPIV) recorded at an 800 Hz rate, which is sufficient to resolve the spatio-temporal dynamics of the flow field and it was combined with simultaneous accelerometer measurements of the airfoil, which was free to oscillate in a direction perpendicular to the freestream. Analysis of the flow field spectra and a Proper Orthogonal Decomposition (POD) of the TRDPIV data of the temporally resolved planar flow fields indicate that the LEB effectively modified the flow field surrounding the airfoil and increased the convecting vortices stand-off distance for over half of the airfoil chord length. It is shown that LEB also causes a redistribution of the flow field spectral energy over a larger range of frequencies.     

Output response identification in a multistable system for piezoelectric energy harvesting

In this paper we examine in detail the multiple responses of a novel vibrational energy harvester composed of a vertical bistable beam whose complex non-linear behavior is tuned via magnetic interaction. The beam was excited horizontally by a harmonic inertial force while mechanical vibrational energy is converted to electrical power through a piezoelectric element. The bistable laminate beam coupled to the piezoelectric transducer showed a variety of complex responses in terms of the beam displacement and harvested power output. The range of vibration patterns in this non-linear system included single-well oscillations and snap-through vibrations of periodic and chaotic character. Harvested power was found to be strongly dependent on the vibration pattern with nonlinearities providing a broadband response for energy harvesting. Wavelet analysis of measured voltage, displacement and velocity time histories indicated the presence of a variety of nonlinear periodic and also chaotic phenomena. To measure the complexity of response time series we applied phase portraits and determine stroboscopic points and multiscale entropy. It is demonstrated that by changing parameters such as the magnetic interaction, the characteristics of the bistable laminate harvester, such as the natural frequency, bandwidth, vibration response and peak power can be readily tailored for harvesting applications.     

Conditions for T<Superscript>2</Superscript> resistivity from electron-electron scattering

Many complex oxides (including titanates, nickelates and cuprates) show a regime in which resistivity follows a power law in temperature (ρ ∝ T ²). By analogy to a similar phenomenon observed in some metals at low temperature, this has often been attributed to electron-electron (Baber) scattering. We show that Baber scattering results in a T ² power law only under several crucial assumptions which may not hold for complex oxides. We illustrate this with sodium metal (ρ _el?el ∝ T ²) and strontium titanate (ρ _el?el \(\hbox{$\not\propto$}\) T ²). We conclude that an observation of ρ ∝T ² is not always sufficient evidence for electron-electron scattering.