A synchrotron computed tomography dataset for validation of longitudinal tensile failure models based on fibre break and cluster development.

We performed in-situ tensile tests on two carbon fibre/epoxy composites with continuous scanning using synchrotron computed tomography (CT). Both composites were cross-ply laminates, and two specimens were tested for each composite. The voxel size was sufficiently small to recognize individual fibres and fibre breaks. For each test, 16-19 volumes were reconstructed, cropped down to the 0° plies and analysed to track fibre break and cluster development. This dataset provides the last CT volume before failure for each of the four specimens as well as the individual fibre break locations in all reconstructed volumes. These data are then plotted against predictions from six state-of-the-art strength models. The target is that these data become a benchmark for the development of new models, inspiring researchers to set up refined experiments and develop improved models.

An approach to the elucidation of metabolic breakdown products of the luteinizing hormone-releasing hormone.

Metabolic breakdown of the luteinizing hormone-releasing hormone (LH-RH) could lead to the following fragments containing pyroglutamic acid: pyroglutamic acid (1), pGlu-His (2), pGLu-His-Trp (3), pGlu-His-Trp-Ser (4), etc., and finally pGlu-His-Trp-Ser-Tyr-Gly-Leu-Arg-Pro-Gly (10). We have synthesized fragments 2-10 and successfully separated all ten metabolites and LH-RH by high-performance liquid chromatography (HPLC) with a muBondapak C18 column. In a test of the viability of the method, cochromatography of fragments 1-10 and LH-RH with the products of chymotryptic digestion of tritiated LH-RH showed radioactive peaks corresponding to the expected products, fragments 3 and 5. Analysis of the products of incubation of a rat kidney homogenate supernatant with LH-RH showed fragments 1-4 and LH-RH. The finding of breakdown at position 4 uncovers a new site of LH-RH breakdown and points the way to the design of potential LH-RH antagonists and agonists where the 4 position would be substituted with unnatural amino acids to prevent breakdown.

Fluorescence nanotomography using resonance energy transfer: demonstration with a protein-sugar complex.

A new approach to structural sensing, fluorescence resonance energy transfer nanotomography, which interprets fluorescence decay measurement in terms of site density analysis of molecular distributions, has been applied to a glucose sensor based on competitive binding with malachite green labelled dextran to the sugar binding protein concanavalin A labelled with allophycocyanin. Opportunities for structural sensing in clinical medicine are highlighted.

A time-resolved near-infrared fluorescence assay for glucose: opportunities for trans-dermal sensing.

We report a time-resolved near-infrared fluorescence assay for glucose detection that incorporates pulsed diode laser excitation. Reduction in fluorescence resonance energy transfer to a malachite green-Dextran complex from allophycocyanin bound to concanavalin A (ConA) due to displacement of the complex by glucose from ConA provides the basis of the assay. The fluorescence quenching kinetics are analysed and discussed in detail. The change in fluorescence decay kinetics in the presence of glucose is found from dimensionality studies to be brought about by a change in the distribution of malachite green-Dextran acceptors. Glucose concentrations are measured in solution to within +/- 10% over the range 0-30 mM.

Potential for ammonia recapture by farm woodlands: design and application of a new experimental facility.

There has been increasing pressure on farmers in Europe to reduce the emissions of ammonia from their land. Due to the current financial climate in which farmers have to operate, it is important to identify ammonia control measures that can be adopted with minimum cost. The planting of trees around farmland and buildings has been identified as a potentially effective and low-cost measure to enhance ammonia recapture at a farm level and reduce long-range atmospheric transport. This work assesses experimentally what fraction of ammonia farm woodlands could potentially remove from the atmosphere. We constructed an experimental facility in southern Scotland to simulate a woodland shelterbelt planted in proximity to a small poultry unit. By measuring horizontal and vertical ammonia concentration profiles within the woodland, and comparing this to the concentration of an inert tracer (SF6) we estimate the depletion of ammonia due to dry deposition to the woodland canopy. Together with measurements of mean ammonia concentrations and throughfall fluxes of nitrogen, this information is used to provide a first estimate of the fraction of emitted ammonia that is recaptured by the woodland canopy. Analysis of these data give a lower limit of recapture of emitted ammonia, at the experimental facility, of 3%. By careful design of shelterbelt woodlands this figure could be significantly higher.

Molecular distribution sensing in a fluorescence resonance energy transfer based affinity assay for glucose.

A newly developed method for determining molecular distribution functions is applied to a widely researched glucose affinity sensor. The reduction in fluorescence resonance energy transfer (FRET) to a malachite green (MG)-dextran complex from allophycocyanin (APC) bound to concanavalin A (ConA) due to displacement of the complex by glucose from ConA provides the basis of the assay. The higher sensitivity and specificity of a new approach to fluorescence decay analysis, over the methods based on conventional Forster-type models, is demonstrated and critical parameters in competitive binding FRET sensing derived.

Coronary risk factor prevalence in a high incidence area: results from the Belfast MONICA Project.

Northern Ireland remains at the top of the world mortality league for ischaemic heart disease. The Province is providing a centre for the World Health Organisation's MONICA Project. Registration of coronary heart disease events began in 1983 and the first of three population surveys took place in 1983-4. A total of 2,361 men and women aged 25-64 years was screened. Subjects were shorter and heavier than their fellow citizens in Great Britain. The estimated mean cholesterol levels in the 25-64-year-old population (5.80 mmol/l in males and 5.85 mmol/l in females) were similar to those reported from Great Britain. Although mean systolic blood pressures were lower, mild diastolic hypertension was considerably more common; cigarette smoking levels were similar. The results were consistent with those expected for an area with a high coronary heart disease mortality, with more than 80% of subjects being at increased risk in terms of the three major factors (cigarette smoking, hypertension and raised cholesterol). Public concern about coronary heart disease has grown and recently the Department of Health and Social Services (NI) has launched a 10-year prevention programme which will primarily employ a population approach.

In-situ analysis of pectic polysaccharides in seed mucilage and at the root surface of Arabidopsis thaliana.

Pectic polysaccharides are a complex set of macromolecules of the primary cell wall matrix with distinct structural domains. The biosynthesis, organisation and function of these domains within cell wall matrices are poorly understood. An immersion immunofluorescence labelling technique was developed for the in-situ analysis of pectic polysaccharides at the surface of seeds and seedlings of Arabidopsis thaliana (L.) Heynh., and used to investigate the occurrence of pectic homogalacturonan (HG) and rhamnogalacturonan-I (RG-I) epitopes. Seed mucilage appeared to consist of two regions: a highly methyl-esterified HG was a major component throughout the mucilage, while an inner region with relatively low porosity was stabilized by calcium-based HG cross-linking. The small size and transparency of Arabidopsis roots allowed the occurrence of pectic HG and RG-I epitopes at root surfaces to be directly determined on whole-mount preparations. Pectic epitopes were not distributed evenly over root surfaces and were notably absent from lateral root apices and from the surface of root hairs. The use of defined antibody probes in the immersion immunolabelling protocol will be useful for the analysis of the influence of growth conditions and genetic factors on pectic polysaccharides in Arabidopsis.

Temporal and spatial regulation of pectic (1-->4)-beta-D-galactan in cell walls of developing pea cotyledons: implications for mechanical properties.

Modifications in cell wall pectic polysaccharides are thought to influence cell-cell adhesion and the mechanical properties of plant tissues. Monoclonal antibodies to epitopes occurring in homo- galacturonan and side chains of rhamnogalacturonan I have been used in an immunolocalization study of cell wall architecture of developing pea cotyledons. Pectic (1-->4)-beta-D-galactan appears in cotyledon cell walls at a defined stage late in development (approximately 26-30 days after anthesis), whereas homogalacturonan and pectic (1-->5)-alpha-L-arabinan are present in cotyledon cell walls throughout development. (1-->4)-beta-galactan was restricted to a distinct thin layer at the plasma membrane face of the cell wall. Anion exchange and immunoaffinity chromatography indicated that the (1-->4)-beta-galactan was associated with acidic pectic components. Mechanical compressive testing of pea cotyledons, before and after (1-->4)-beta-galactan appearance, indicated that the cotyledons with the galactan-rich cell wall layer were twice as firm as those with no detectable (1-->4)-beta-galactan.

Near-infrared fluorescence lifetime assay for serum glucose based on allophycocyanin-labeled concanavalin A.

We describe an assay scheme for glucose based on fluorescence resonance energy transfer (FRET) between concanavalin A (con A), labeled with the near-infrared fluorescent protein allophycocyanin (APC) as donor, and dextran labeled with malachite green (MG) as acceptor. Glucose competitively displaces dextran-MG and leads to reduction in FRET, assessed by time-domain fluorescence lifetime measurements using time-correlated single-photon counting. The assay is operative in the glucose concentration range 2.5-30 mM, and therefore suitable for use in monitoring diabetes control. Albumin and serum inhibit FRET but the interference can be prevented by removal of high molecular weight substances by membrane filters. APC shows promise for incorporation in an implanted glucose sensor which can be interrogated from outside the body.

Pectin: cell biology and prospects for functional analysis.

Pectin is a major component of primary cell walls of all land plants and encompasses a range of galacturonic acid-rich polysaccharides. Three major pectic polysaccharides (homogalacturonan, rhamnogalacturonan-I and rhamnogalacturonan-II) are thought to occur in all primary cell walls. This review surveys what is known about the structure and function of these pectin domains. The high degree of structural complexity and heterogeneity of the pectic matrix is produced both during biosynthesis in the endomembrane system and as a result of the action of an array of wall-based pectin-modifying enzymes. Recent developments in analytical techniques and in the generation of anti-pectin probes have begun to place the structural complexity of pectin in cell biological and developmental contexts. The in muro de-methyl-esterification of homogalacturonan by pectin methyl esterases is emerging as a key process for the local modulation of matrix properties. Rhamnogalacturonan-I comprises a highly diverse population of spatially and developmentally regulated polymers, whereas rhamnogalacturonan-II appears to be a highly conserved and stable pectic domain. Current knowledge of biosynthetic enzymes, plant and microbial pectinases and the interactions of pectin with other cell wall components and the impact of molecular genetic approaches are reviewed in terms of the functional analysis of pectic polysaccharides in plant growth and development.

Upper-limb function in Australian children with traumatic brain injury: A controlled, prospective study.

OBJECTIVE: To describe upper-limb function in children with mild and severe traumatic brain injury (TBI), by using both quantitative and qualitative measures. DESIGN: Controlled, prospective cohort study with assessment points initially, at 6 months, and at 2 years after TBI. SETTING: A tertiary pediatric trauma center in Australia. PATIENTS: Fifty-one children, ranging in age up to 14 years, who were consecutive admissions with TBI. On the basis of initial and persisting abnormal coma score and persistence of posttraumatic amnesia, they were assigned to either a mild (n = 26) or a severely injured (n = 25) group. Thirty children admitted with non-TBI trauma were recruited as a control group. MAIN OUTCOME MEASURES: Quantitative measures included Bruininks-Oseretsky Test of Motor Proficiency and Peabody Developmental Motor Scales. Qualitative measures included Brunnstrom Recovery Stages (adapted), categoric scales of muscle tone, grasp used when handwriting, quality of writing product, bilateral activity, and splint use. RESULTS: There was little difference between the groups on the standardized assessments for subjects who could complete the tests. Qualitative measures showed the severe TBI group to have more difficulties with gross arm control, hand control, and hand function. CONCLUSION: Children with severe TBI experience more and persisting difficulties with upper-limb function. It is essential to include both quantitative and qualitative measures in this type of research.

Carvedilol, a novel cardiovascular agent, inhibits development of vascular and ventricular hypertrophy in spontaneously hypertensive rats.

The effects of carvedilol, a novel cardiovascular agent, were evaluated in developing spontaneously hypertensive rats (SHR) for effects on hemodynamics, and the ability to effect the development of left ventricular, and vascular hypertrophy associated with chronic hypertension. Chronic oral administration of low dose carvedilol (20 mg/kg/day) was initiated when rats were 5 weeks of age, and experiments progressed until 14 weeks of age. Carvedilol-treated SHR had significantly reduced systolic blood pressures and heart rates throughout the duration of the experiment, and had significantly reduced ventricle/body weights by approximately 9.0%. Morphologic analysis of tertiary branches of the mesenteric artery revealed that carvedilol-treated SHR had significant reductions in medial cross-sectional area. Carvedilol produced concentration-dependent inhibition of basal [3H]thymidine incorporation in cultured SHR vascular smooth muscle cells, as well as by stimulation produced by PDGF (1 nM), EDGF (1 nM), thrombin (0.5 U/ml), or endothelin-1 (1 nM), indicating that carvedilol had direct anti-mitogenic activity. The present studies demonstrate that low dose carvedilol produced sustained reductions in blood pressure and heart rate in developing SHR that were accompanied by significant inhibition in the development of vascular and myocardial hypertrophy. The morphological changes induced by carvedilol may be mediated by a combination of hemodynamic effects, as well as by direct anti-mitogenic effects on vascular smooth muscle.

The Westmead Pediatric TBI Multidisciplinary Outcome study: use of functional outcomes data to determine resource prioritization.

OBJECTIVE: To measure functional outcome in the 2 years after traumatic brain injury (TBI) in 2 groups of children and to determine the usefulness of a TBI severity classification system for resource allocation. DESIGN: Prospective inception cohort study with 3 assessment points during the 2 years after trauma. SETTING: Tertiary pediatric trauma center in Sydney, Australia. PARTICIPANTS: Eighty-one consecutive admissions aged 0 to 14 years. Fifty-one were allocated to the Mild (n = 26) or Severe (n = 25) TBI groups, according to preset determinants of severity; 30 admissions with non-TBI trauma constituted the control group. MAIN OUTCOME MEASURES: Standardized psychometric and clinical assessments of cognition, communication and feeding ability, motor performance (ambulation, fine and gross motor), neurologic status, self-care independence, and school/academic performance. RESULTS: Those with Mild TBI severity had no significant deficits at the 2-year data point. In contrast, those in the Severe TBI group demonstrated continued problems with fine motor performance, neurologic status, self care, and school/academic performance. CONCLUSIONS: A classification system has been developed that may be useful in the allocation of children with a TBI, age younger than 15 years, to 1 of 2 severity groups early in their rehabilitation. This classification system may be useful in determining areas of high and low resource prioritization.