Investigation of fibrillin microfibrils in the canine cruciate ligament in dogs with different predispositions to ligament rupture.

Cranial cruciate ligament disease (CCLD) is the most common cause of pelvic limb lameness in dogs but its precise aetiopathogenesis is uncertain. Fibrillin microfibrils (FM) are complex macro-molecular assemblies found in many tissues including ligaments, where they are thought to play an important mechanical role. We hypothesised that FM ultrastructural variation correlates with the differing predisposition of canine breeds to CCLD. Non-diseased cranial and caudal cruciate ligaments (CCLs and CaCLs) were obtained from Greyhound (GH) and Staffordshire Bull Terrier (SBT) cadavers. Fibrillin microfibrils were extracted from the ligaments by bacterial collagenase digestion, purified by size-exclusion chromatography and subsequently visualized by atomic force microscopy (AFM). With AFM, FMs have a characteristic beads-on-a-string appearance. For each FM, periodicity (bead-bead distance) and length (number of beads/FM) was measured. Fibrillin microfibril length was found to be similar for GH and SBT, with non-significant inter-breed and inter-ligament differences. Fibrillin microfibril periodicity varied when comparing GH and SBT for CCL (GH 60.2 ± 1.4 nm; SBT 56.2 ± 0.8 nm) and CaCL (GH 55.5 ± 1.6 nm; SBT 61.2 ± 1.2 nm). A significant difference was found in the periodicity distribution when comparing CCL for both breeds (P < 0.00001), further, intra-breed differences in CCL vs CaCL were statistically significant within both breeds (P < 0.00001). The breed at low risk of CCLD exhibited a periodicity profile which may be suggestive of a repair and remodelling within the CCL.

The role of subcutaneous tissue stiffness on microneedle performance in a representative in vitro model of skin.

There has been growing interest in the mechanical behaviour of skin due to the rapid development of microneedle devices for drug delivery applications into skin. However, most in vitro experimentation studies that are used to evaluate microneedle performance do not consider the biomechanical properties of skin or that of the subcutaneous layers. In this study, a representative experimental model of skin was developed which was comprised of subcutaneous and muscle mimics. Neonatal porcine skin from the abdominal and back regions was used, with gelatine gels of differing water content (67, 80, 88 and 96%) to represent the subcutaneous tissue, and a type of ballistic gelatine, Perma-Gel®, as a muscle mimic. Dynamic nanoindentation was used to characterize the mechanical properties of each of these layers. A custom-developed impact test rig was used to apply dense polymethylmethacrylate (PMMA) microneedles to the skin models in a controlled and repeatable way with quantification of the insertion force and velocity. Image analysis methods were used to measure penetration depth and area of the breach caused by microneedle penetration following staining and optical imaging. The nanoindentation tests demonstrated that the tissue mimics matched expected values for subcutaneous and muscle tissue, and that the compliance of the subcutaneous mimics increased linearly with water content. The abdominal skin was thinner and less stiff as compared to back skin. The maximum force decreased with gel water content in the abdominal skin but not in the back skin. Overall, larger and deeper perforations were found in the skin models with increasing water content. These data demonstrate the importance of subcutaneous tissue on microneedle performance and the need for representative skin models in microneedle technology development.

Anion replacement at Au(110)/electrolyte interfaces.

A characteristic reflection anisotropy spectrum (RAS) is observed from a Au(110) surface in a wide range of electrolytes and combinations of pH and applied potentials. It is suggested that this common RAS profile arises from an interaction between the potential applied to the Au(110) electrode and the dipole moments of oxidized species that locates the Fermi level at a common position with respect to the electronic band structure of Au. Rapid changes in this RAS profile are observed for Au(110)/H2SO4 as the potential is switched between 0.3 V and 0.6 V, a potential range in which the surface is not reconstructed and below the potential range of surface oxidation. The spectral changes are completed in less than 10 ms, are reversible and are attributed to the replacement of adsorbed anions by an oxygenated species.

The stability of the Au(1 1 0)-(1 × 3) surface reconstruction in electrochemical environments.

Changes in the reflection anisotropy (RAS) profile of the Au(1 1 0)-(1 × 3)/Na2SO4 interface over 25 h are attributed to the slow accumulation of impurities on the Au(1 1 0) surface which reduce the intensity of a transition involving a surface state that makes a positive contribution to the RAS profile at 1.8 eV. The growth in the intensity of a feature that makes a negative contribution to the RAS profile at 2.6 eV and the reduction in the intensity of contributions to higher energy is attributed to shifts in the energy of the surface band structure relative to the Fermi level caused by the accumulation of impurities. There is no clear explanation of the subsequent decay of the 2.6 eV feature or the long term reduction in intensity to high energy of the RAS profile.

Localized micro- and nano-scale remodelling in the diabetic aorta.

Diabetes is strongly associated with cardiovascular disease, but the mechanisms, structural and biomechanical consequences of aberrant blood vessel remodelling remain poorly defined. Using an experimental (streptozotocin, STZ) rat model of diabetes, we hypothesized that diabetes enhances extracellular protease activity in the aorta and induces morphological, compositional and localized micromechanical tissue remodelling. We found that the medial aortic layer underwent significant thickening in diabetic animals but without significant changes in collagen or elastin (abundance). Scanning acoustic microscopy demonstrated that such tissue remodelling was associated with a significant decrease in acoustic wave speed (an indicator of reduced material stiffness) in the inter-lamellar spaces of the vessel wall. This index of decreased stiffness was also linked to increased extracellular protease activity (assessed by semi-quantitative in situ gelatin zymography). Such a proteolytically active environment may affect the macromolecular structure of long-lived extracellular matrix molecules. To test this hypothesis, we also characterized the effects of diabetes on the ultrastructure of an important elastic fibre component: the fibrillin microfibril. Using size exclusion chromatography and atomic force microscopy, we isolated and imaged microfibrils from both healthy and diabetic aortas. Microfibrils derived from diabetic tissues were fragmented, morphologically disrupted and weakened (as assessed following molecular combing). These structural and functional abnormalities were not replicated by in vitro glycation. Our data suggest that proteolysis may be a key driver of localized mechanical change in the inter-lamellar space of diabetic rat aortas and that structural proteins (such as fibrillin microfbrils) may be biomarkers of diabetes induced damage.

Raman phonon emission in a driven double quantum dot.

The compound semiconductor gallium-arsenide (GaAs) provides an ultra-clean platform for storing and manipulating quantum information, encoded in the charge or spin states of electrons confined in nanostructures. The absence of inversion symmetry in the zinc-blende crystal structure of GaAs however, results in a strong piezoelectric interaction between lattice acoustic phonons and qubit states with an electric dipole, a potential source of decoherence during charge-sensitive operations. Here we report phonon generation in a GaAs double quantum dot, configured as a single- or two-electron charge qubit, and driven by the application of microwaves via surface gates. In a process that is a microwave analogue of the Raman effect, phonon emission produces population inversion of the two-level system and leads to rapid decoherence of the qubit when the microwave energy exceeds the level splitting. Comparing data with a theoretical model suggests that phonon emission is a sensitive function of the device geometry.

Optical reflectance anisotropy of the growth of Fe monolayers on W(110).

We report measurements of the optical anisotropy of Fe layers grown on the W(110) surface using reflection anisotropy spectroscopy (RAS). As the first monolayer of Fe is deposited onto W(110), the resonance-like RAS profile of the clean surface is reduced in intensity. We find evidence for the surface state on W(110) surviving as an interface state following Fe deposition. We observe an anisotropic optical response from Fe layers grown on top of the first two monolayers, where a broad peak at 3 eV dominates the RAS response. The results are simulated in terms of a layered Fresnel reflection model incorporating either a strained Fe overlayer or an Fe overlayer whose dielectric properties are approximated by a simple Lorentzian oscillator. Both approaches are found to produce simulated RA spectra that are in good agreement with experiment. The former approach provides evidence that RAS can detect anisotropy in strained overlayers and that 7 ML films have bulk-like electronic and optical properties.

Continuous measurement of a microwave-driven solid state qubit.

We analyze the dynamics of a continuously observed, damped, microwave-driven solid state charge qubit, consisting of a single electron in a double well potential. The microwave field induces transitions between the qubit eigenstates, which have a profound effect on the detector output current. Useful information about the qubit dynamics, such as dephasing and relaxation rates, and the Rabi frequency, can be extracted from the detector conductance and output noise power spectrum. We also propose a technique for single-shot electron spin readout, for spin based quantum information processing, which has a number of practical advantages over existing schemes.

Population inversion of a driven two-level system in a structureless bath.

We derive a master equation for a driven double quantum dot damped by an unstructured phonon bath, and calculate the spectral density. We find that bath-mediated photon absorption is important at relatively strong driving, and may even dominate the dynamics, inducing population inversion of the double-dot system. This phenomenon is consistent with recent experimental observations.

The structure of the mixed OH + H2O overlayer on Pt[111].

The structure of the mixed p(3 x 3)-(3OH + 3H2O) phase on Pt[111] has been investigated by low-energy electron diffraction-IV structure analysis. The OH + H2O overlayer consists of hexagonal rings of coplanar oxygen atoms interlinked by hydrogen bonds. Lateral shifts of the O atoms away from atop sites result in different O-O separations and hexagons with only large separations (2.81 and 3.02 angstroms) linked by hexagons with alternating separations of 2.49 and 2.813.02 angstroms. This unusual pattern is consistent with a hydrogen-bonded network in which water is adsorbed in cyclic rings separated by OH in a p(3 x 3) structure. The top-most two layers of the Pt atoms relax inwards with respect to the clean surface and both show vertical buckling of up to 0.06 angstroms. In addition, significant shifts away from the lateral bulk positions have been found for the second layer of Pt atoms.

Selective spin coupling through a single exciton.

We present a novel scheme for performing a conditional phase gate between two spin qubits in adjacent semiconductor quantum dots through delocalized single exciton states, formed through the interdot Förster interaction. We consider two resonant quantum dots, each containing a single excess conduction band electron whose spin embodies the qubit. We demonstrate that both the two-qubit gate and arbitrary single-qubit rotations may be realized to a high fidelity with current semiconductor and laser technology.

Continuous quantum measurement: inelastic tunneling and lack of current oscillations.

We study the dynamics of a charge qubit, consisting of a single electron in a double well potential coupled to a point-contact (PC) electrometer, using the quantum trajectories formalism. Contrary to previous predictions, we show formally that, in the sub-Zeno limit, coherent oscillations in the detector output are suppressed, and the dynamics is dominated by inelastic processes in the PC. Furthermore, these reduce the detector efficiency and induce relaxation even when the source-drain bias is zero. This is of practical significance since it means the detector will act as a source of decoherence. Finally, we show that the sub-Zeno dynamics is divided into two regimes: low and high bias in which the PC current power spectra show markedly different behavior.

A software tool to straighten curved chromosome images.

A software tool for straightening curved chromosomes has been developed and integrated into the freely available image analysis application Image SXM (available via the Internet at http://reg.ssci.liv.ac.uk). This new tool straightens curvilinear objects in one simple step after minimal input from the user. The ends of a curvilinear chromosome are identified by the user using the mouse and a window is opened displaying the object as it would appear if it was straightened out. This image processing produces linear images of chromosomes with no loss of resolution or spatial calibration, making subsequent analysis significantly more straightforward.

CI-1017, a functionally M1-selective muscarinic agonist: design, synthesis, and preclinical pharmacology.

The five muscarinic receptor subtypes (M1-M5) are characterized by seven helices that define a transmembrane cavity which serves as the binding pocket for agonists and antagonists. The five cavities appear to be topographically different enough to permit subtype selectivity among antagonists but not among classical agonists which tend to be smaller in size than antagonists. It was reasoned that synthesis of muscarinic agonists longer/larger than their classical counterparts might result in subtype selectivity. M1 subtype selectivity was found in a class of 1-azabicyclo[2.2.1]heptan-3-one, O-(3-aryl-2-propynyl) oximes. One of these, CI-1017, improved spatial memory of hippocampally deficient mice and nbM-lesioned rats at doses of 1.0-3.2 and 0.1-0.3 mg/kg, respectively, while producing parasympathetic side effects only at very high doses (100-178 mg/kg). Additionally, CI-1017 inhibited production of amyloidogenic A beta and increased secretion of soluble APP. Thus, CI-1017, besides treating AD symptomatically, may also retard its progression. CI-1017 has recently completed phase I clinical trials.

Blockade of the MAP kinase pathway suppresses growth of colon tumors in vivo.

The mitogen-activated protein kinase pathway is thought to be essential in cellular growth and differentiation. Here we report the discovery of a highly potent and selective inhibitor of the upstream kinase MEK that is orally active. Tumor growth was inhibited as much as 80% in mice with colon carcinomas of both mouse and human origin after treatment with this inhibitor. Efficacy was achieved with a wide range of doses with no signs of toxicity, and correlated with a reduction in the levels of activated mitogen-activated protein kinase in excised tumors. These data indicate that MEK inhibitors represent a promising, noncytotoxic approach to the clinical management of colon cancer.

Design and synthesis of m1-selective muscarinic agonists: (R)-(-)-(Z)-1-Azabicyclo[2.2.1]heptan-3-one, O-(3-(3'-methoxyphenyl)-2-propynyl)oxime maleate (CI-1017), a functionally m1-selective muscarinic agonist.

The synthesis and SAR of a series of (Z)-(+/-)-1-azabicyclo[2.2. 1]heptan-3-one, O-(3-aryl-2-propynyl)oximes are described. The biochemistry and pharmacology of 24Z (PD 142505) and its enantiomers are highlighted. 24Z is functionally an m1-selective muscarinic agonist. Efficacy and m1 selectivity reside in the R enantiomer, (R)-24Z (CI-1017).

Photoemission Study of CO Adsorption on Gd.

Angle-resolved UV photoemission spectroscopy has been used to investigate the interaction of CO with Gd films, grown on W(110). The results suggest the CO absorbs dissociatively, initially forming Gd(2)O(3), with subsequent catalytic oxidation of CO to form carbonate.

Utilization of Photoemission Cross-Section Effects for Monitoring Thin-Film Growth in UHV.

Given the tunability of synchrotron radiation, photoemission spectroscopy can be used to monitor the growth modes of thin films in UHV. Cross-section effects such as Cooper minima and resonant photoemission can be exploited to maximize the adsorbate signal and minimize that from the substrate. Under favourable circumstances growth can be monitored in real time at coverages of <1%. As an example the growth of Gd on W(110) is investigated.