Biomedical subjects

R E Palmer

Publications and source records attributed to R E Palmer.

At least 19 recordsLinked to original sources

PubMed2007

Broad spectrum of Pompe disease in patients with the same c.-32-13T->G haplotype.

BACKGROUND: Pompe disease (acid maltase deficiency, glycogen storage disease type II; OMIM 232300) is an autosomal recessive lysosomal storage disorder characterized by acid alpha-glucosidase deficiency due to mutations in the GAA gene. Progressive skeletal muscle weakness affects motor and respiratory functions and is typical for all forms of Pompe disease. Cardiac hypertrophy is an additional fatal symptom in the classic infantile subtype. c.-32-13T-->G is the most common mutation in adults. OBJECTIVE: To delineate the disease variation among patients with this mutation and to define the c.-32-13T-->G haplotypes in search for genotype-phenotype correlations. METHODS: We studied 98 compound heterozygotes with a fully deleterious mutation (11 novel mutations are described) and the common c.-32-13T-->G mutation. RESULTS: All patients were Caucasian. None had the classic infantile form of Pompe disease. The clinical course varied far more than anticipated (age at diagnosis <1 to 78 years; age at onset: <1 to 52 years). The acid alpha-glucosidase activities in a subset of patients ranged from 4 to 19.9 nmol/mg/h. Twelve different c.-32-13T-->G haplotypes were identified based on 17 single-nucleotide polymorphisms located in the GAA gene. In 76% of the cases, c.-32-13T-->G was encountered in the second most common GAA core haplotype (DHRGEVVT). In only one case was c.-32-13T-->G encountered in the major GAA core haplotype (DRHGEIVT). CONCLUSION: Patients with the same c.-32-13T-->G haplotype (c.q. GAA genotype) may manifest first symptoms at different ages, indicating that secondary factors may substantially influence the clinical course of patients with this mutation.

Adolescent↗

PubMed2006

Pinning of size-selected Pd nanoclusters on graphite.

The production of stable cluster arrays on smooth surfaces has several potential technological applications. We report a study of the pinning of size-selected palladium nanoclusters on the graphite surface. The clusters formed during gas aggregation in vacuum are projected with sufficient kinetic energy to create a defect in the graphite surface. The energy necessary to create such an immobilizing defect is investigated as a function of the palladium cluster size. The palladium pinning energy is found to deviate from the simple binary collision model as appropriate to previously reported silver and gold results. This finding is in agreement with the deviation of nickel clusters and points to the influence of the interatomic cluster bonding on the mechanics of the collision.

Journal Article↗

PubMed2006

Bioluminescence of monolayers of firefly luciferase immobilized on graphite.

We report on the immobilization of the firefly protein luciferase on the hydrophobic surface of graphite. Observation by liquid-phase atomic force microscopy of islands with a height consistent with the size of a single molecule confirmed that the protein was contained within a monomolecular layer. The enzyme activity was assayed by single-photon counting of the bioluminescence, which is the catalytic product of luciferase. Attachment to the surface modified the efficiency of the enzyme, but the introduction of the substrates luciferin and ATP resulted in the reactivation of the enzyme. The functionalized graphite surface was employed as a cathode in a bioelectrochemical cell. This demonstrated that the electric field caused a substantial loss of enzyme catalytic activity.

Adenosine Triphosphate↗

PubMed2005

Two-electron dissociation of single molecules by atomic manipulation at room temperature.

Using the tip of a scanning tunnelling microscope (STM) to mechanically manipulate individual atoms and molecules on a surface is now a well established procedure. Similarly, selective vibrational excitation of adsorbed molecules with an STM tip to induce motion or dissociation has been widely demonstrated. Such experiments are usually performed on weakly bound atoms that need to be stabilized by operating at cryogenic temperatures. Analogous experiments at room temperature are more difficult, because they require relatively strongly bound species that are not perturbed by random thermal fluctuations. But manipulation can still be achieved through electronic excitation of the atom or molecule by the electron current tunnelling between STM tip and surface at relatively high bias voltages, typically 1-5 V. Here we use this approach to selectively dissociate chlorine atoms from individual oriented chlorobenzene molecules adsorbed on a Si(111)-7 x 7 surface. We map out the final destination of the chlorine daughter atoms, finding that their radial and angular distributions depend on the tunnelling current and hence excitation rate. In our system, one tunnelling electron has nominally sufficient energy to induce dissociation, yet the process requires two electrons. We explain these observations by a two-electron mechanism that couples vibrational excitation and dissociative electron attachment steps.

Journal Article↗

PubMed2005

Electrostatically stabilised nanoparticles: self-organization and electron-beam patterning.

The self-organisation of citrate- and magnesium oleate-stabilised gold nanoparticles on SiO2/Si substrates was investigated. In drop deposition, nucleation of citrate-stabilised gold nanoparticles was observed at the rim of the droplet, symmetric or multibranched dendroid gold structures were found in the area between the rim and the central part of the droplet, depending on the drying temperature. Homogeneous submonolayer nanoparticle coverage was obtained by immersion of amineterminated SiO2/Si surfaces into a citrate-stabilised colloidal gold acidic solution. Drop deposition of magnesium oleate-stabilised gold nanoparticles onto the SiO2/Si surfaces resulted in the formation of uniformly close-packed nanoparticle arrays. Under electron beam irradiation, no apparent changes were found for monolayer films of citrate-stabilized particles, but sintering of the nanoparticles was observed in multilayer films. In contrast, coalescence of magnesium oleate-stabilised gold nanoparticle occurred in monolayer films after electron irradiation.

Citric Acid↗

PubMed2004

Decoration of surfaces with size-selected clusters and molecular manipulation at room temperature: precision and uncertainty in organizing atoms.

The deposition onto surfaces of clusters of atoms, prepared and size-selected in the gas phase, is, like atomic or molecular manipulation with the scanning tunnelling microscope, an appealing (but parallel) route to the creation of nanoscale surface features. Both of these seemingly orthogonal approaches allow, in principle, a selected number of atoms to be organized, and both are strongly affected by the lateral thermal diffusion of the constituent atoms, molecules or clusters over the surface. In this sense, the room-temperature (as opposed to cryogenic-temperature) regime can be regarded as a hostile environment for organizing atoms. In this paper we review recent achievements in size-selected cluster deposition and molecular manipulation at room temperature and thus address the fundamental question: with what precision can we organize atoms at room temperature?

Journal Article↗

PubMed2004

Deposition of size-selected metal clusters generated by magnetron sputtering and gas condensation: a progress review.

This paper presents a topical review of the production and deposition of size-selected metal clusters generated by magnetron sputtering and gas condensation. Clusters with up to 75,000 atoms can be obtained by controlling the gas pressure in the sputtering source, prior to size selection with a novel (time-of-flight) mass filter. The clusters are deposited on the model graphite substrate to study the cluster-surface interaction. Two regimes for cluster deposition have been identified at elevated impact energies: (i) 'pinning' (ca. 10 eV per atom) and (ii) implantation (ca. 100 eV per atom). Of particular importance is the pinning regime, since this allows the fabrication of monodispersed cluster arrays, which are stable against diffusion at room temperature (and above). The deposition of size-selected metal clusters represents a novel method of preparing surface nanostructures, with potential applications including model catalyst studies and the immobilization of biological molecules.

Journal Article↗

PubMed2003

Insight into the partial oxidation of propene: the reactions of 2-propen-1-ol on clean and O-covered Mo(110).

The reactions of 2-propen-1-ol (allyl alcohol) were studied on clean and O-covered Mo(110) to understand the effect of resonance stabilization and the presence of surface oxygen on reaction selectivity. Propene is the only gaseous hydrocarbon product evolved from allyl alcohol reaction on O-covered Mo(110). Water and dihydrogen are also produced, along with a small amount of adsorbed carbon. We estimated, using X-ray photoelectron spectroscopy, that approximately 70% of the 0.11 ML of 2-propen-1-ol that reacts forms propene. In contrast, the dominant reaction pathway on the clean surface is nonselective decomposition to adsorbed carbon and hydrogen, leading to a 23% selectivity for propene formation. On both clean and O-covered Mo(110), X-ray photoelectron spectroscopy and infrared spectroscopy identify allyloxy as the reaction intermediate yielding propene. These results are discussed in the context of propene oxidation and periodic trends in reactivity.

Journal Article↗

PubMed2003

Mechanisms of molecular manipulation with the scanning tunneling microscope at room temperature: chlorobenzene/si(111)-(7 x 7).

We report a systematic experimental investigation of the mechanism of desorption of chlorobenzene molecules from the Si(111)-(7 x 7) surface induced by the STM at room temperature. We measure the desorption probability as a function of both tunneling current and a wide range of sample bias voltages between -3 V and +4 V. The results exclude field desorption, thermally induced desorption, and mechanical tip-surface effects. They indicate that desorption is driven by the population of negative (or positive) ion resonances of the chemisorbed molecule by the tunneling electrons (or holes). Density functional calculations suggest that these resonant states are associated with the pi orbitals of the benzene ring.

Journal Article↗

PubMed2003

Scaling relations for implantation of size-selected Au, Ag, and Si clusters into graphite.

The deposition of size-selected clusters represents a new route to the fabrication of truly nanometer-scale surface architectures, e.g., nanopores. We report a systematic experimental study, coupled with molecular dynamics simulations, of the implantation depths of size-selected Au7, Ag7, and Si7 clusters in the model graphite substrate. For impact energies between 1.0 and 5.5 keV, we find that the implantation depth scales linearly with the momentum of the clusters for all three types of cluster. This "universal" behavior is consistent with a (viscous) retarding force proportional to the velocity of the cluster, akin to Stokes's law.

Journal Article↗

PubMed2003

Nanostructured surfaces from size-selected clusters.

The deposition of ionized beams of size-selected atomic clusters onto well-defined substrates represents a new method of preparing nanostructured surfaces, with lateral feature sizes in the range 1-10 nm. 'Pinning' of the incident clusters prevents the diffusion of the clusters on the surface, and thus preserves the gas-phase cluster size, even at room temperature and above. At the same time, advances in diblock copolymer techniques allow the preparation of ordered two-dimensional arrays of clusters. Here we discuss the creation and applications of these nanostructured surfaces, ranging from the fabrication of semiconductor nanostructures to the immobilization of protein molecules.

Manufactured Materials↗

PubMed2002

Fabrication of ordered arrays of InP microstructures by wet chemical etching with Au masks.

Ordered arrays of InP microstructures have been fabricated on InP(001) substrates by wet chemical etching in aqueous HCl with patterned Au masks. The masks were produced by Au deposition through copper grids or a monolayer of polystyrene microspheres. Square InP mesas (20 x 20 microns) and pillars (approximately 100 nm in both diameter and height) were both produced and characterized by scanning electron microscopy and atomic force microscopy.

Crystallization↗

PubMed2001

WT1 regulates the expression of the major glomerular podocyte membrane protein Podocalyxin.

The WT1 tumor suppressor gene encodes a zinc finger transcription factor expressed in differentiating glomerular podocytes. Complete inactivation of WT1 in the mouse leads to failure of mesenchymal induction and renal agenesis, an early developmental phenotype that prevents analysis of subsequent stages in glomerular differentiation [1]. In humans with Denys-Drash Syndrome, a heterozygous germline mutation in WT1 is associated with specific defects in glomeruli and an increased risk for developing Wilms Tumor [2,3]. WT1 target genes implicated in cell cycle regulation and cellular proliferation have been proposed [4], but the link between WT1 function and glomerular differentiation is unexplained. Here, we show that inducible expression of WT1 in rat embryonic kidney cell precursors leads to the induction of endogenous Podocalyxin, the major structural membrane protein of glomerular podocytes, which is implicated in the maintenance of filtration slits. Binding of WT1 to conserved elements within the Podocalyxin gene promoter results in potent transcriptional activation, and the specific expression pattern of Podocalyxin in the developing kidney mirrors that of WT1 itself. These observations support a role for WT1 in the specific activation of a glomerular differentiation program in renal precursors and provide a molecular basis for the glomerulonephropathy that is characteristic of Denys-Drash Syndrome.

3T3 Cells↗

PubMed2001

Cascades of transcriptional induction during human lymphocyte activation.

Lymphocyte activation is known to be associated with the induction of genes implicated in cytokine signaling and cellular proliferation. High-density microarrays offer the means to monitor global cellular expression profiles, temporal relationships between classes of transcripts, and alterations associated with human disease or immunosuppression. We sought to determine whether microarray analysis would accurately reflect the normal pattern of gene expression following human T cell activation, and whether the complex expression patterns identified could be analyzed to produce a functional profile of lymphocyte activation. We examined a time course of sequential expression profiles for 6,800 cellular transcripts in human lymphocytes activated with concanavalin A. Expression patterns were grouped using clustering analysis and validated using Northern blotting. Genes known to be induced following T cell activation were accurately identified, and the qualitative patterns of gene expression were well correlated between Northern and microarray analyses. Quantitative differences in gene expression levels were less well correlated between these two techniques. Expression profile analysis revealed the sequential induction of groups of functionally similar genes, whose temporal coregulation underscores known cellular events during T cell activation. This functional "fingerprint" of lymphocyte activation may prove useful for comparisons of lymphocyte responses under experimental conditions and in disease states.

Cluster Analysis↗

PubMed2001

Miniature heart cell force transducer system implemented in MEMS technology.

A fully submersible force transducer system for use with isolated heart cells has been implemented using microelectromechanical systems (MEMS) technology. By using integrated circuit fabrication techniques to make mechanical as well as electrical components, the entire low-mass transducer is only a few cubic millimeters in size and is of higher fidelity (approximately 100 nN and 13.3 kHz in solution) than previously available. When chemically activated, demembranated single cells attached to the device contract and slightly deform a strain gauge whose signal is converted to an amplified electrical output. When integrated with a video microscope, the system is capable of optical determination of contractile protein striation periodicity and simultaneous measurement of heart cell forces in the 100-nN to 50-microN range. The average measured maximal force was Fmax = 5.77 +/- 2.38 microN. Normalizing for the cell's cross-sectional area, Fmax/area was 14.7 +/- 7.7 mN/mm2. Oscillatory stiffness data at frequencies up to 1 kHz has also been recorded from relaxed and contracted cells. This novel MEMS force transducer system permits higher fidelity measurements from cardiac myocytes than available from standard macro-sized transducers.

Animals↗

PubMed1999

A diazo-2 study of relaxation mechanisms in frog and barnacle muscle fibres: effects of pH, MgADP, and inorganic phosphate.

The aim of this study was to compare the effects of increased concentrations of MgADP, inorganic phosphate (Pi) and H+ ([MgADP], [Pi] and [H+], respectively) on the rate of relaxation in two different muscle types: skinned muscle fibres from the frog Rana temporaria and myofibrillar bundles from the giant Pacific acorn barnacle Balanus nubilus. Relaxation transients are produced by the photolysis of diazo-2 and are well fitted with a double exponential curve, giving two rate constants: k1 [5.6+/-0.1 s-1 for barnacle, n=30; 26.3+/-0.7 s-1 for frog, n=14 (mean+/-SEM)] and k2 [0.6+/-0.1 s-1 in barnacle, n=30; 10.4+/-1.0 s-1 in frog, n=14 (mean+/-SEM)], at 10 degrees C. Decreasing the pH by 0.5 pH units did not significantly affect k1 for barnacle relaxation [5.6+/-0.1 s-1 (mean+/-SEM), n=15] compared to the decrease in k1 of 40% seen in frog. Use of the Ca2+-sensitive fluorescent label acrylodan on barnacle wild-type troponin C demonstrated that decreasing the pH from 7.0 to 6.6 only alters the pCa50 value by 0.23 in the cuvette, while stopped-flow experiments with acrylodan revealed no significant change in koff from the labelled protein [322+/-32 s-1 at pH 7.0 and 381+/-24 s-1 (mean+/-SEM) at pH 6.6]. Increasing [MgADP] by 20 microM (50 microM added ADP) from control values of 50 microM in frog decreased k1 to 12.3+/-0.4 s-1 (mean+/-SEM, n=8), and at 400 microM MgADP, k1=9.6+/-0.1 s-1 (mean+/-SEM, n=12). In barnacle, 500 microM MgADP had a much smaller effect on k1 (4.0+/-0. 9 s-1, mean+/-SEM, n=8). Increasing the free [Pi] from the contaminant level of 0.36 mM to 1.9 mM slowed k1 by approximately 15% in barnacle [4.8+/-0.8 s-1, mean+/-SEM, n=7], compared to a approximately 30% reduction seen in frog. We conclude that the differences between barnacle and frog seen here are most probably due to different isomers of the contractile proteins, and that events underlying the crossbridge cycle are the same or similar. We interpret our results according to a model of crossbridge transitions during relaxation.

2-Naphthylamine↗

PubMed1999

The effect of phosphate on the relaxation of frog skeletal muscle.

The effect of phosphate on the relaxation of isometrically contracting single skinned fibres from the semitendinosus muscle of the frog Rana temporaria has been investigated using laser pulse photolysis of the photolabile caged calcium-chelator diazo-2 to rapidly reduce the Ca2+ (<2 ms) within the fibre and produce >90% relaxation of force. Relaxation occurred in two phases - an initial linear shoulder which lasted approximately 20 ms followed by a double-exponential phase which gave two rate constants, k1 (43.4+/-1. 8 s-1, mean +/-SEM, n=14) and k2 (15.6+/-0.3 s-1, mean +/-SEM, n=14) at 12 degreesC. Increased phosphate concentrations did not affect the linear phase, but slowed the double-exponential phase following photolysis of diazo-2 in a dose-dependent fashion (k50= 0.9 mM for k1, 1.15 mM for k2). Reducing the concentration of contaminating phosphate (from 640 microM to 100 microM) led to an increase in the rate of the double-exponential phase (k1=67.1+/-4.4 s-1, k2=19.7+/-0. 6 s-1, mean +/-SEM, n=12). Time-resolved measurements of sarcomere length during relaxation, both in control fibres and in the presence of a raised phosphate concentration, reveal a <2% change throughout the whole relaxation transient, and less than 0.1% at the end of the linear phase. This finding implies that gross changes in sarcomere length do not contribute to the decay of the relaxation transient seen upon diazo-2 photolysis. Our results suggest that cross-bridges in states prior to phosphate release are already committed to force generation and must relax by releasing phosphate, rather than by a reversal of the force-generating step to a weakly bound, low-force phosphate-bound state. These findings also indicate that an increase in the phosphate concentration within muscle fibres plays an important part in the slowing of relaxation observed in skeletal muscle fatigue and that the relaxation transients observed upon diazo-2 photolysis represent a disengagement of the cross-bridges.

Adenosine Diphosphate↗

PubMed1998

Joint injuries of the hand in athletes.

The variety of injuries that occur in the joints of the hand range from skin abrasions to displaced intra-articular fractures. This article discusses serious injuries involving ligaments, volar plate, and bones at specific anatomic regions of the joints of the hand.

Athletic Injuries↗