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Biomedical subjects

E J Williams

Publications and source records attributed to E J Williams.

At least 19 recordsLinked to original sources
PubMed2006

Comparison of ultraviolet absorbance, chemiluminescence, and DOAS instruments for ambient ozone monitoring.

This paper evaluates the accuracy of ozone measurements made by monitors that determine ozone concentrations in ambient air by UV absorption. These monitors are typically used to measure ozone for the purpose of establishing local compliance to air-quality standards. The study was predicated by the concern that commercially available UV absorbance O3 monitors may be subject to interference from volatile organic carbon (VOC) species that absorb light at 254 nm. To test for these and other effects, we compared simultaneous O3 measurements made by a commercial UV O3 monitor with an O3-NO chemiluminescence instrument, which is not subject to interference by VOC compounds. The comparisons were carried out in the summers of 1999 and 2000 at urban/industrial sites in Nashville and Houston, and in 2004 aboard a ship in the Gulf of Maine. In the two urban areas, we also compared the 03 measurements from these two methods with O3 measurements made by a long-path differential optical absorption spectrometer (DOAS). Our tests indicate that, with well-maintained monitors, there are no significant interferences even in areas with significant ambient concentrations of potentially interfering VOCs.

Environmental Monitoring↗
PubMed2005

A review of troponin assay performance in Wales: can the same (method-dependent) decision limits be used in different sites?

An audit of troponin measurement protocols in use in Wales showed significant variations in practice with no consensus on analytical methods, or in the selection of a decision limit for the diagnosis of myocardial damage. Peer review data on assay imprecision at concentrations approaching the analytical limit of detection are lacking. The objective of the study was to establish clinically relevant precision profiles for the troponin methods used throughout Wales, which could be used to develop a standardized approach to the selection of a decision limit. A series of five pools of human serum spiked with troponin I-T complex were prepared and stored at -70 degrees C until dispatch. Five sets of each pool were dispatched to all Welsh laboratories and stored at -20 degrees C until analysis. The analysis protocol consisted of two replicates of each pool per batch, and two batches per day for five days (n=20). All the laboratories performed all the measurements in the same week. The lowest concentration providing a 10% coefficient of variation (CV) was 0.02 microg/L for the Roche method and 0.11 microg/L for the Beckman AccuTnI. The lowest concentrations that could be distinguishable from the 99th percentile reference limit were 0.02 microg/L for the Roche method and 0.07 microg/L for the Beckman AccuTnI. Current methods do not achieve the 10% CV at the 99th percentile reference limit proposed by the European Society of Cardiology. Use of the lowest concentration that can be reliably distinguished from the 99th percentile reference limit offers a novel alternative decision limit, which provides a slightly lower concentration than at the 10% CV but maintaining confidence in the assay that false-positive rates will be minimized.

Chemistry, Clinical↗
PubMed2005

Recommended standards for biochemical markers of myocardial damage: All Wales Clinical Biochemistry Audit Group.

The effective use of cardiac-specific troponin estimations in the diagnosis of acute myocardial infarction (AMI) is clouded by the imprecise definition surrounding the decision limits. This has led to a wide variation of criteria for the diagnosis of myocardial infarction. A survey of troponin measurements in Welsh laboratories, undertaken in 2003 under the auspices of the All Wales Clinical Biochemistry Audit Group, revealed significant variations in laboratory and clinical practice. Extensive discussion and consultation led by a working group of clinical biochemists and cardiologists in Wales culminated in recommendations concerning the use of troponin assays to establish myocardial damage. The key recommendations are: Cardiac troponin (T or I) should be the first-line test for myocardial damage; Two samples should be collected, at admission and 12-24 h later. The first sample is used for 'rule in' purposes, but not to 'rule out' myocardial damage; Only one threshold (cut-off) value for troponin should be quoted on laboratory reports, values above which are indicative of myocardial damage. A study by the Wales External Quality Assurance Scheme (WEQAS) enabled the derivation of the recommended cut-off concentrations of troponin for defining myocardial damage, defined for each assay as the concentration that can be reliably distinguished, with a confidence interval of 99%, from the 99th percentile reference limit. These recommended standards provide a rationale for a uniform approach for troponin assays for patients with chest pain, working towards a standardized approach to the diagnosis and management of patients presenting with acute coronary syndromes.

Biomarkers↗
PubMed2003

Influence of baseline hematocrit on between-subject BOLD signal change using gradient echo and asymmetric spin echo EPI.

The dependence of BOLD signal change (BSC) on baseline hematocrit is in the process of being characterized, primarily using conventional Gradient Echo (GE) echo planar imaging (EPI). We describe the first empiric exploration of this relationship using, in addition to GE, Spin Echo (SE) and two Asymmetric Spin Echo EPI sequences (ASE10 and ASE20), which are less susceptible to large vessel noise. Motor cortex BSC was measured (N = 17) and regressed against hematocrit and hemoglobin concentration using linear and non-linear functions. GE measurements of BSC yielded a positive linear relationship (r(2) = 0.240, p = 0.0459) whereas a positive non-linear relationship was observed using ASE10 (r(2) = 0.571, p = 0.0146). Results suggest that between-subjects BSC is significantly dependent on baseline hematocrit. The nature of dependence, and implications for quantitative studies vary with the vessel size selectivity of the imaging sequence, and with the effect of hematocrit on blood viscosity in the imaged vessels.

Brain Mapping↗
PubMed2002

Severe panniculitis caused by homozygous ZZ alpha1-antitrypsin deficiency treated successfully with human purified enzyme (Prolastin).

Severe panniculitis caused by alpha1-antitrypsin deficiency is very rare even though the ZZ phenotype occurs in 1 : 3500 of the population of northern Europe. We describe a 33-year-old woman with rapidly progressing panniculitis and extensive skin necrosis with multiple life-threatening complications. Initial treatment followed by maintenance therapy with human purified enzyme (Prolastin, Bayer, Bridgend, U.K.) has been life-saving.

Adult↗
PubMed2001

Identification of an N-cadherin motif that can interact with the fibroblast growth factor receptor and is required for axonal growth.

In this study, we show that the neurite outgrowth response stimulated by N-cadherin is inhibited by a recently developed and highly specific fibroblast growth factor receptor (FGFR) antagonist. To test whether the N-cadherin response also requires FGF function, we developed peptide mimetics of the receptor binding sites on FGFs. Most mimetics inhibit the neurite outgrowth response stimulated by FGF in the absence of any effect on the N-cadherin response. The exceptions to this result were two mimetics of a short FGF1 sequence, which has been shown to interact with the region of the FGFR containing the histidine-alanine-valine motif. These peptides inhibited FGF and N-cadherin responses with similar efficacy. The histidine-alanine-valine region of the FGFR has previously been implicated in the N-cadherin response, and a candidate interaction site has been identified in extracellular domain 4 of N-cadherin. We now show that antibodies directed to this site on N-cadherin inhibit the neurite outgrowth response stimulated by N-cadherin, and peptide mimetics of the site inhibit N-cadherin and FGF responses. Thus, we can conclude that N-cadherin contains a novel motility motif in extracellular domain 4, and that peptide mimetics of this motif can interact with the FGFR.

Amino Acid Motifs↗
PubMed2001

Effect of slice orientation on reproducibility of fMRI motor activation at 3 Tesla.

The effect of slice orientation on reproducibility and sensitivity of 3T fMRI activation using a motor task has been investigated in six normal volunteers. Four slice orientations were used; axial, oblique axial, coronal and sagittal. We applied analysis of variance (ANOVA) to suprathreshold voxel statistics to quantify variability in activation between orientations and between subjects. We also assessed signal detection accuracy in voxels across the whole brain by using a finite mixture model to fit receiver operating characteristic (ROC) curves to the data. Preliminary findings suggest that suprathreshold cluster characteristics demonstrate high motor reproducibility across subjects and orientations, although a significant difference between slice orientations in number of activated voxels was demonstrated in left motor cortex but not cerebellum. Subtle inter-orientation differences are highlighted in the ROC analyses, which are not obvious by ANOVA; the oblique axial slice orientation offers the highest signal detection accuracy, whereas coronal slices give the lowest.

Analysis of Variance↗
PubMed2001

Local similarity in evolutionary rates extends over whole chromosomes in human-rodent and mouse-rat comparisons: implications for understanding the mechanistic basis of the male mutation bias.

The sex chromosomes and autosomes spend different times in the germ line of the two sexes. If cell division is mutagenic and if the sexes differ in number of cell divisions, then we expect that sequences on the X and Y chromosomes and autosomes should mutate at different rates. Tests of this hypothesis for several mammalian species have led to conflicting results. At the same time, recent evidence suggests that the chromosomal location of genes on autosomes affects their rate of evolution at synonymous sites. This suggests a mutagenic source different from germ cell replication. To correctly interpret the previous estimates of male mutation bias, it is crucial to understand the degree and range of this local similarity. With a carefully chosen randomization protocol, local similarity in synonymous rates of evolution can be detected in human-rodent and mouse-rat comparisons. However, the synonymous-site similarity in the mouse-rat comparison remains weak. Simulations suggest that this difference between the mouse-human and the mouse-rat comparisons is not artifactual and that there is therefore a difference between humans and rodents in the local patterns of mutation or selection on synonymous sites (conversely, we show that the previously reported absence of a local similarity in nonsynonymous rates of evolution in the human-rodent comparison was a methodological artifact). We show that linkage effects have a long-range component: not one in a million random genomes shows such levels of autosomal heterogeneity. The heterogeneity is so great that more autosomes than expected by chance have rates of synonymous evolution comparable with that of the X chromosome. As autosomal heterogeneity cannot be owing to different times spent in the germ line, this demonstrates that the dominant determiner of synonymous rates of evolution is not, as has been conjectured, the time spent in the male germ line.

Animals↗
PubMed2001

Relaxin inhibits effective collagen deposition by cultured hepatic stellate cells and decreases rat liver fibrosis in vivo.

BACKGROUND: Following liver injury, hepatic stellate cells (HSC) transform into myofibroblast-like cells (activation) and are the major source of type I collagen and the potent collagenase inhibitors tissue inhibitors of metalloproteinases 1 and 2 (TIMP-1 and TIMP-2) in the fibrotic liver. The reproductive hormone relaxin has been reported to reduce collagen and TIMP-1 expression by dermal and lung fibroblasts and thus has potential antifibrotic activity in liver fibrosis. AIMS: To determine the effects of relaxin on activated HSC. METHODS: Following isolation, HSC were activated by culture on plastic and exposed to relaxin (1-100 ng/ml). Collagen deposition was determined by Sirius red dye binding and radiolabelled proline incorporation. Matrix metalloproteinase (MMP) and TIMP expression were assessed by zymography and northern analysis. Transforming growth factor beta1 (TGF-beta1) mRNA and protein levels were quantified by northern analysis and ELISA, respectively. RESULTS: Exposure of activated HSC to relaxin resulted in a concentration dependent decrease in both collagen synthesis and deposition. There was a parallel decrease in TIMP-1 and TIMP-2 secretion into the HSC conditioned media but no change in gelatinase expression was observed. Northern analysis demonstrated that primary HSC, continuously exposed to relaxin, had decreased TIMP-1 mRNA expression but unaltered type I collagen, collagenase (MMP-13), alpha smooth muscle actin, and TGF-beta1 mRNA expression. CONCLUSION: These data demonstrate that relaxin modulates effective collagen deposition by HSC, at least in part, due to changes in the pattern of matrix degradation.

Animals↗
PubMed2001

Selective inhibition of fibroblast growth factor (FGF)-stimulated mitogenesis by a FGF receptor-1-derived phosphopeptide.

The activated fibroblast growth factor receptor (FGFR)-1 is phosphorylated on five tyrosine residues outside the catalytic site. Although one such residue, Tyr730, is flanked by potential binding sites for phosphotyrosine-interacting molecules, a physiological role for this region is still controversial. We report that a cell-permeant phosphopeptide mimic of this site, FGFR730(p)Y, inhibits FGF-mediated mitogenesis in cells with no effect on responses stimulated by other growth factors. A similar phosphopeptide corresponding to the phospholipase Cgamma binding site on the receptor had no effect on the mitogenic response. The FGFR730(p)Y peptide did not inhibit phosphorylation of p90/FRS2 or Erk, suggesting that it does not act by inhibiting the Erk-kinase cascade. However, the FGFR730(p)Y peptide bound Shc in a manner requiring both phosphorylated tyrosine and a putative PTB domain binding determinant. These data suggest that the peptide might inhibit mitogenesis by competing with the corresponding site on the FGFR for the ability to bind SHC.

3T3 Cells↗
PubMed2000

Covariation of GC content and the silent site substitution rate in rodents: implications for methodology and for the evolution of isochores.

Many attempts to test selectionist and neutralist models employ estimates of synonymous (Ks) and non-synonymous (Ka) substitution rates of orthologous genes. For example, a stronger Ka-Ks correlation than expected under neutrality has been argued to indicate a role for selection and the absence of a Ks-GC4 correlation has been argued to be inconsistent with neutral models for isochore evolution. However, both of these results, we have shown previously, are sensitive to the method by which Ka and Ks are estimated. Using a maximum likelihood (ML) estimator (GY94) we found a positive correlation between Ks and GC4 and only a weak correlation between Ka and Ks, lower than expected under neutral expectations. This ML method is computationally slow. Recently, a new ad hoc approximation of this ML method has been provided (YN00). This is effectively an extension of Li's protocol but that also allows for codon usage bias. This method is computationally near-instantaneous and therefore potentially of great utility for analysis of large datasets. Here we ask whether this method might have such applicability. To this end we ask whether it too recovers the two unusual results. We report that when the ML and earlier ad hoc methods disagree, YN00 recovers the results described by the ML methods, i.e. a positive correlation between GC4 and Ks and only a weak correlation between Ks and Ka. If the ML method can be trusted, then YN00 can also be considered an adequately reliable method for analysis of large datasets. Assuming this to be so we also analyze further the patterns. We show, for example, that the positive correlation between GC4 and Ks is probably in part a mutational bias, there being more methyl induced CpG-->TpG mutations in GC rich regions. As regards the evolution of isochores, it seems inappropriate to use the claimed lack of a correlation between GC and Ks as definitive evidence either against or for any model. If the positive correlation is real then, we argue, this is hard to reconcile with the biased gene conversion model for isochore formation as this predicts a negative correlation.

Animals↗
PubMed2000

The proteins of linked genes evolve at similar rates.

Much more variation in the rate of protein evolution occurs than is expected by chance. But why some proteins evolve rapidly but others slowly is poorly resolved. It was proposed, for example, that essential genes might evolve slower than dispensable ones, but this is not the case; and despite earlier claims, rates of evolution do not correlate with amino-acid composition. A few patterns have been found: proteins involved in antagonistic co-evolution (for example, immune genes, parasite antigens and reproductive conflict genes) tend to be rapidly evolving, and there is a correlation between the rate of protein evolution and the mutation rate of the gene. Here we report a new highly statistically significant predictor of a protein's rate of evolution, and show that linked genes have similar rates of protein evolution. There is also a weaker similarity of rates of silent site evolution (see ref. 13), which appears to be, in part, a consequence of the similarity in rates of protein evolution. The similarity in rates of protein evolution is not a consequence of underlying mutational patterns. A pronounced negative correlation between the rate of protein evolution and a covariant of the recombination rate indicates that rates of protein evolution possibly reflect, in part, the local strength of stabilizing selection.

Animals↗
PubMed2000

The molecular evolution of signal peptides.

Signal peptides direct mature peptides to their appropriate cellular location, after which they are cleaved off. Very many random alternatives can serve the same function. Of all coding sequences, therefore, signal peptides might come closest to being neutrally evolving. Here we consider this issue by examining the molecular evolution of 76 mouse-rat orthologues, each with defined signal peptides. Although they do evolve rapidly, they evolve about half as fast as neutral sequences. This indicates that a substantial proportion of mutations must be under stabilizing selection. A few putative signal sequences lack a hydrophobic core and these tend to be more slowly evolving than others, indicating even stronger stabilizing selection. However, closer scrutiny suggests that some of these represent mis-annotations in GenBank. It is also likely that some of the substitutions are not neutral. We find, for example, that the rate of protein evolution correlates with that of the mature peptide. This may be a result of compensatory evolution. We also find that signal peptides of immune genes tend to be faster evolving than the average, which suggests an association with antagonistic co-evolution. Previous reports also indicated that the signal peptide of the imprinted gene, Igf2r, is also unusually fast evolving. This, it was hypothesized, might also be indicative of antagonistic co-evolution. Comparison of Igf2r's signal peptide evolution shows that, although it is not an outlier, its rate of evolution is comparable to that of many of the faster evolving immune system signal sequences and 5/6 of the amino acid changes do not conserve hydrophobicity. This is at least suggestive that there is something unusual about Igf2r's signal sequence.

Amino Acid Substitution↗
PubMed2000

Distribution of the interleukin-8 receptors, CXCR1 and CXCR2, in inflamed gut tissue.

There is increasing evidence to suggest that the potent neutrophil chemoattractant interleukin-8 (IL-8) has an important role in the pathogenesis of inflammatory bowel disease. IL-8 mediates its actions via two cell surface receptors, CXCR1 and CXCR2. This paper describes the distribution of these IL-8 receptors in the normal gastrointestinal tract and how this is modified in ulcerative colitis (UC). Paraffin-embedded colonic resection specimens were stained with monoclonal antibodies directed against CXCR1 and CXCR2 in ten cases of total UC, 16 cases of appendicitis, and 11 histologically normal sections. A semiquantitative scale of 0-4 was used to assess the proportion and intensity of positively stained cells within certain defined areas of tissue. A comparative assessment was made of the distribution of various cell populations. Dual immunostaining was used to confirm the phenotype of positively staining cells. In the histologically normal colon, the antibody against CXCR1 stained a subpopulation of macrophages deep to the epithelium and germinal centre lymphocytes. A similar pattern of staining was seen in acute appendicitis, with in addition some positively stained neutrophil polymorphs. In UC, there was up-regulation of CXCR1, with a striking increase in positively stained macrophages throughout the mucosa and of B and T lymphocytes outside the germinal centre areas. There was also intense up-regulation of CXCR1 expression by the luminal epithelium, reflected in the epithelial staining score (mean+/-SE=1.8+/-0.44 for UC cases, vs. 0.23+/-0.16 for controls and 0.25+/-0.14 for acute appendicitis). CXCR2 was only expressed on a small population of lamina propria mononuclear cells and crypt epithelial cells, with no significant differences observed between the groups. These results suggest that IL-8 may, through CXCR1, have a role beyond neutrophil recruitment in mediating the immune response in UC and that this is not merely a consequence of the acute inflammation seen in UC.

Acute Disease↗
PubMed2000

INP, a novel N-cadherin antagonist targeted to the amino acids that flank the HAV motif.

The classical cadherins are homophilic binding molecules that play fundamental roles in several biological processes, including axonal growth and synaptic plasticity. The structures of the amino-terminal homophilic binding domains of N-cadherin and E-cadherin have been resolved. However, the mechanisms that govern cadherin binding and specificity remain contentious. In the present study we have used a peptide competition approach to probe for small linear determinants of cadherin binding. We demonstrate that a linear peptide mimetic of a short sequence in ECD1 of N-cadherin (INPISGQ) functions as a highly specific and potent antagonist of N-cadherin function with an IC(50) value of approximately 15 microM. Peptide mimetics of the corresponding motif in chick R-cadherin also inhibited N-cadherin function, albeit with lower efficacy. In contrast, peptide mimetics of the corresponding motif in E- or P-cadherin failed to inhibit N-cadherin function. A short cyclic peptide that contained only the INP motif from N-cadherin was also a potent N-cadherin antagonist (IC(50) approximately 15 microM). Analysis of existing crystal structures suggests that the peptides are likely to antagonize N-cadherin function by binding to the region that flanks the HAV motif at the adhesion dimer interface.

3T3 Cells↗
PubMed2000

CAMs and axonal growth: a critical evaluation of the role of calcium and the MAPK cascade.

Calcium has long been recognized as a key player in the control of axonal growth and guidance. Recent studies lend support to this pivotal role by showing that local changes in calcium can directly induce the formation of filopodia in vivo and turn a growth cone in vitro. Under normal growth conditions, the L1 adhesion molecule has now been shown to induce local rather than global changes in calcium in growth cones, and this suggests that cell adhesion molecules (CAMs) use localized calcium transients to stimulate axonal growth and guidance. A number of recent reports have demonstrated that the neurite outgrowth response stimulated by L1 and other adhesion molecules (NCAM, N-cadherin, laminin) also depends in part upon the integrity of the MAPK cascade in cells. In this review we consider the recent data and suggest that calcium and the MAPK cascade might be required for very distinct growth cone functions. Finally, we will consider the contentious issue of how the above CAMs activate signaling cascades in growth cones and review the recently available data that support the hypothesis that at least one of these CAMs (N-cadherin) might promote growth cone motility by directly interacting with the FGFR in growth cones.

Animals↗
PubMed2000

Increased expression of connective tissue growth factor in fibrotic human liver and in activated hepatic stellate cells.

BACKGROUND/AIMS: Connective tissue growth factor is a recently described mitogenic protein implicated in a variety of fibrotic disorders. Connective tissue growth factor may be a downstream mediator of the pro-fibrotic and mitogenic actions of transforming growth factor-beta, promoting extracellular matrix deposition and fibrogenesis. As transforming growth factor-beta is considered important to the pathogenesis of hepatic fibrosis, we examined the possible contribution of connective tissue growth factor to this process. METHODS: Connective tissue growth factor expression was examined in normal and fibrotic human and rat livers using RT-PCR and ribonuclease protection assays, and in primary cultures of rat hepatic stellate cells by Northern and Western blotting. RESULTS: Ribonuclease protection assays demonstrated connective tissue growth factor mRNA was increased 3-5-fold in human fibrotic liver compared with normal. RT-PCR showed this mRNA was increased in carbon-tetrachloride-treated rat liver. Northern analysis showed connective tissue growth factor mRNA was increasingly expressed during progressive activation of cultured rat hepatic stellate cells. Western analysis confirmed that freshly isolated hepatic stellate cells secreted relatively little connective tissue growth factor compared with hepatic stellate cells activated in culture. Hepatic stellate cells stimulated with transforming growth factor-beta showed increased expression of connective tissue growth factor mRNA and protein. CONCLUSIONS: Connective tissue growth factor mRNA is consistently upregulated in human liver cirrhosis of various aetiologies, supporting a role for this growth factor in hepatic fibrogenesis. Our studies suggest that hepatic stellate cells may be an important source of hepatic connective tissue growth factor in vivo, particularly following stimulation with transforming growth factor-beta.

Animals↗