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R A Walker

Publications and source records attributed to R A Walker.

At least 19 recordsLinked to original sources
PubMed2007

The aged breast.

There is a clear association between the development of breast cancer and increasing age, with 80% of cancers occurring in women more than 50 years of age and one-third in women over 70 years. Following the menopause the breast undergoes involution, with the main changes affecting the terminal ductal lobular unit. There is an increase in oestrogen receptor alpha (ERalpha)-positive cells, a decrease in proliferation but, in comparison to premenopausal breasts, a greater number of ERalpha-proliferating cells. The breast cancers that occur in women >/= 75 years are more likely to be ER-positive, with a low growth rate and limited expression of HER-2 and p53. It is proposed that uneven involution of the breast, the persistence of at-risk lesions, the presence of ERalpha-proliferating cells and local oestrogen metabolism in breast adipose tissue are factors in the development of breast cancers with a well-differentiated phenotype.

Adipose Tissue↗
PubMed2006

Quantification of immunohistochemistry--issues concerning methods, utility and semiquantitative assessment I.

Immunohistochemistry is no longer a technique used only for research but is employed increasingly for diagnosis and for the assessment of therapeutic biomarkers. The latter, in particular, often require a semiquantitative evaluation of the extent of their presence. There are many factors that can affect this that relate to the method: fixation of tissue, duration and type of antigen retrieval, antibody specificity, antibody dilution and detection systems. Other complexities relate to assessment. Different scoring systems are used for either the same or different antigens. Cut-off levels for assessing whether a tissue is 'positive' or 'negative' can vary for the same antigen. Whilst there are quality assurance schemes for the methodology that have improved standards of staining, there are no similar schemes that relate to interpretation, although errors here can create as many problems. There have been improvements in automated analysis but availability is limited and it is still predominantly a research tool. In order for quantification of immunohistochemistry to be a reliable and reputable tool, there must be easy to use, reproducible, standardized protocols for assessment which are international. Improvements in automated analysis with wider applicability could lead to standardization.

Biomarkers↗
PubMed2006

Impact of a national external quality assessment scheme for breast pathology in the UK.

BACKGROUND: This article presents the results and observed effects of the UK National Health Service Breast Screening Programme (NHSBSP) external quality assurance scheme in breast histopathology. AIMS/METHODS: The major objectives were to monitor and improve the consistency of diagnoses made by pathologists and the quality of prognostic information in pathology reports. The scheme is based on a twice yearly circulation of 12 cases to over 600 registered participants. The level of agreement was generally measured using kappa statistics. RESULTS: Four main situations were encountered with respect to diagnostic consistency, namely: (1) where consistency is naturally very high-this included diagnosing in situ and invasive carcinomas (and certain distinctive subtypes) and uncomplicated benign lesions; (2) where the level of consistency was low but could be improved by making guidelines more detailed and explicit-this included histological grading; (3) where consistency could be improved but only by changing the system of classification-this included classification of ductal carcinoma in situ; and (4) where no improvement in consistency could be achieved-this included diagnosing atypical hyperplasia and reporting vascular invasion. Size measurements were more consistent for invasive than in situ carcinomas. Even in cases where there is a high level of agreement on tumour size, a few widely outlying measurements were encountered, for which no explanation is readily forthcoming. CONCLUSIONS: These results broadly confirm the robustness of the systems of breast disease diagnosis and classification adopted by the NHSBSP, and also identify areas where improvement or new approaches are required.

Breast Neoplasms↗
PubMed2006

The NHS breast screening programme (pathology) EQA: experience in recent years relating to issues involved in individual performance appraisal.

BACKGROUND: The original role of the National Health Service breast screening programme (pathology) external quality assessment (EQA) scheme was educational; it aimed to raise standards, reinforce use of common terminology, and assess the consistency of pathology reporting of breast disease in the UK. AIMS/METHODS: To examine the performance (scores) of pathologists participating in the scheme in recent years. The scheme has evolved to help identify poor performers, reliant upon setting an acceptable cutpoint. Therefore, the effects of different cutpoint strategies were evaluated and implications discussed. RESULTS/CONCLUSIONS: Pathologists who joined the scheme improved over time, particularly those who did less well initially. There was no obvious association between performance and the number of breast cancer cases reported each year. This is not unexpected because the EQA does not measure expertise, but was established to demonstrate a common level of performance (conformity to consensus) for routine cases, rather than the ability to diagnose unusual/difficult cases. A new method of establishing cutpoints using interquartile ranges is proposed. The findings also suggest that EQA can alter a pathologist's practice: those who leave the scheme (for whatever reason) have, on average, marginally lower scores. Consequently, with the cutpoint methodology currently used (which is common to several EQA schemes) there is the potential for the cutpoint to drift upwards. In future, individuals previously deemed competent could subsequently be erroneously labelled as poor performers. Due consideration should be given to this issue with future development of schemes.

Breast Neoplasms↗
PubMed2005

Surface charge effects on solvation across liquid/liquid and model liquid/liquid interfaces.

Resonance enhanced second harmonic generation (SHG) coupled with novel solvatochromic surfactants has been used to explore solvation across liquid/liquid interfaces in the presence of excess surface charge in the aqueous phase. The surfactants--dubbed "molecular rulers"--consist of hydrophobic, solvatochromic chromophores connected to charged headgroups via variable length alkyl spacers. Interfacial dipolar width is monitored as a function of chromophore/headgroup separation. Data show that cationic and anionic surfactants of equivalent lengths sample very different environments across a water/cyclohexane interface. The effective excitation wavelengths of cationic surfactants are shifted persistently to higher energies than in bulk cyclohexane while anionic surfactants solvation converges smoothly from the aqueous to the organic limit with increasing spacer length. To further evaluate the effect of surface charge on interfacial solvation, SHG was used to probe the environment surrounding molecular ruler chromophores adsorbed to the aqueous liquid/vapor interface in the presence of densely packed monolayers of 1-octanol. These monolayer systems are shown to reproduce qualitatively many of the features associated with bulk water/alkane interfaces and are logistically easier to assemble. Changing the ionic strength of the underlying aqueous sub-phase suggests that the headgroup of cationic molecular ruler surfactants alters the electronic structure of the chromophore rather than properties of the surrounding nonpolar environment.

Journal Article↗
PubMed2005

Perfluorochemicals: potential sources of and migration from food packaging.

Perfluorochemicals are widely used in the manufacturing and processing of a vast array of consumer goods, including electrical wiring, clothing, household and automotive products. Furthermore, relatively small quantities of perfluorochemicals are also used in the manufacturing of food-contact substances that represent potential sources of oral exposure to these chemicals. The most recognizable products to consumers are the uses of perfluorochemicals in non-stick coatings (polytetrafluoroethylene (PTFE)) for cookware and also their use in paper coatings for oil and moisture resistance. Recent epidemiology studies have demonstrated the presence of two particular perfluorochemicals, perfluorooctane sulfonate (PFOS) and perfluorooctanoic acid (PFOA) in human serum at very low part per billion levels. These perfluorochemicals are biopersistent and are the subject of numerous studies investigating the many possible sources of human exposure. Among the various uses of these two chemicals, PFOS is a residual impurity in some paper coatings used for food contact and PFOA is a processing aid in the manufacture of PTFE used for many purposes including non-stick cookware. Little information is available on the types of perfluorochemicals that have the potential to migrate from perfluoro coatings into food. One obstacle to studying migration is the difficulty in measuring perfluorochemicals by routine conventional analytical techniques such as GC/MS or LC-UV. Many perfluorochemicals used in food-contact substances are not detectable by these conventional methods. As liquid chromatography-mass spectrometry (LC/MS) develops into a routine analytical technique, potential migrants from perfluoro coatings can be more easily characterized. In this paper, data will be presented on the types of perfluoro chemicals that are used in food packaging and cookware. Additionally, research will be presented on the migration or potential for migration of these chemicals into foods or food simulating liquids. Results from migration tests show mg kg(-1) amounts of perfluoro paper additives/coatings transfer to food oil. Analysis of PTFE cookware shows residual amounts of PFOA in the low microg kg(-1) range. PFOA is present in microwave popcorn bag paper at amounts as high as 300 microg kg(-1).

Caprylates↗
PubMed2004

Reliability of routine disc susceptibility testing by the British Society for Antimicrobial Chemotherapy (BSAC) method.

OBJECTIVES: To ascertain the agreement between MICs determined at a central laboratory, and susceptible, intermediate and resistant categorizations based on zone diameters recorded at diagnostic laboratories using the BSAC standardized method. METHODS: Standardized disc susceptibility tests were performed at sentinel laboratories in three surveys, with MIC tests performed on the collected isolates at a reference laboratory. The organisms comprised over 3300 Enterobacteriaceae, Acinetobacter spp., pseudomonads, staphylococci and enterococci, with over 29 000 antibiotic/organism tests in total. RESULTS: More than 90% of the antibiotic/organism combinations classed as susceptible by disc tests in the sentinel laboratories were confirmed by MIC testing. Disagreements were more frequent where disc tests indicated resistance, with half of the piperacillin/tazobactam resistance and one-third of the cephalosporin resistance found in Enterobacteriaceae by disc tests not being confirmed, and with three-quarters of teicoplanin resistance in enterococci not confirmed. None of the few apparent cases of meropenem resistance in Enterobacteriaceae or linezolid, quinupristin/dalfopristin or vancomycin resistance in staphylococci were confirmed by MIC testing. When disagreements were found between disc- and MIC-based categorization, MICs were commonly, although not invariably, one to three doubling dilutions above or below the breakpoint. However, many of the disagreements where MICs were three or more dilutions from the breakpoint were not seen when disc tests were repeated in the central laboratory. CONCLUSIONS: The BSAC disc method seems adequate for confirming susceptibility to guide therapy and to monitor resistance trends. Nevertheless, there must be concern about the over-estimation of many resistances, and frequent zone:MIC disagreements for isolates with borderline susceptibility.

Bacteria↗
PubMed2003

Emergence of a fluoroquinolone-resistant strain of Streptococcus pneumoniae in England.

OBJECTIVE: To determine the epidemiological relationship between pneumococci of serotype 9V, with reduced susceptibility to ciprofloxacin, penicillin and erythromycin, referred to the Reference Laboratory during 1997-2001. METHODS: Isolates were characterized by multilocus sequence typing (MLST), PFGE, and sequencing of parC and gyrA. Relevant clinical data were sought. RESULTS: Forty-eight isolates were received from nine laboratories in England, but 35 (73%) were from one laboratory in Birmingham, and were mostly from elderly patients receiving ofloxacin or ciprofloxacin for respiratory infections. There were two quinolone resistance phenotypes, with ciprofloxacin, moxifloxacin and gemifloxacin MICs of 8-32, 0.5-1 and 0.125-0.25 mg/L, and 64-256, 4-16 and 1-4 mg/L, respectively. Each of three isolates from the former group had mutations in parC, whereas each of nine isolates from the more resistant group had mutations in both parC and gyrA. Several also had increased quinolone efflux. Typing of 27 quinolone-resistant isolates showed that eight were indistinguishable from the epidemic Spain9V-3 (ST156) clone, while the remainder belonged to a novel but related type (ST609), that differed from Spain9V-3 at 2/7 alleles (2 bp changes in aroE and 1 bp change in gdh). Both MLST types were represented among isolates with high- and low-level quinolone resistance. Three of five serotype 9V isolates from Birmingham, with reduced susceptibility to penicillin and erythromycin, and ciprofloxacin MICs of 1-2 mg/L, belonged to MLST type ST609, while another was indistinguishable from the Spain9V-3 clone. Review of records of 32 patients from Birmingham indicated that some isolates were nosocomial, whereas others were acquired in the community. CONCLUSIONS: In the late 1990s, a quinolone-resistant strain, clonally related to Spain9V-3, emerged in England, principally in Birmingham.

Aged↗
PubMed2003

Identification of Ncd tail domain-binding sites on the tubulin dimer.

The Drosophila non-claret disjunctional (Ncd) kinesin-like protein is required for spindle assembly in oocytes and spindle maintenance in early embryos. Through the action of ATP-dependent microtubule (MT)-binding sites in the head and ATP-independent MT-binding sites in the tail, Ncd may bundle and, perhaps, slide MTs relative to each other. Our previous work on the MT-binding site of the Ncd tail domain demonstrated that this site, like the MT-binding sites of tau, contains basic residues flanked by proline residues and can promote MT assembly and stability. Here, we characterize the interactions of a monomeric Ncd tail protein with subtilisin-digested MTs in order to identify sites on the tubulin dimer that interact with the Ncd tail. The results provide evidence for four such binding sites per tubulin dimer and support the hypothesis that each binding site consists of a cluster of acidic residues in the C-terminal regions of alpha- and beta-tubulin.

Animals↗
PubMed2003

Primary breast myoepithelial cells exert an invasion-suppressor effect on breast cancer cells via paracrine down-regulation of MMP expression in fibroblasts and tumour cells.

In normal breast and ductal carcinoma in situ, myoepithelial cells form an incomplete layer separating the epithelial compartment from the stromal environment. Transition to invasive disease is marked by penetration of the myoepithelial-basement membrane (BM) interface. One mechanism involved in tumour invasion is breakdown of extracellular matrices by matrix metalloproteinases (MMPs). It was hypothesized that myoepithelial cells may modulate tumour invasion by controlling MMP gene expression, both in tumour cells and in peri-ductal fibroblasts. To investigate this, myoepithelial cells from normal breast were purified and characterized and their effect on tumour cell invasive potential was assessed. The effect on MMP gene expression of breast cancer cells cultured alone or in combination with primary normal breast fibroblasts was also analysed using RT-PCR with ELISA quantitation, with zymographic analysis to measure enzyme activity. Normal breast myoepithelial cells significantly reduced invasion by the breast cancer cell lines MCF-7, T47D, MDA-MB 231, and MDA-MB 468 when they were cultured alone or in the presence of a fibroblast population. Reduced invasion was associated with changes in MMP gene expression. In those tumour cells expressing MMP, there was a significant down-regulation of MMP-2 (MDA-MB 468, p<0.001), MMP-9 (MDA-MB 231, p=0.05; MDA-MB 468, p<0.001), and MT1-MMP (p<0.001 for both MDA-MB 231 and MDA-MB 468). Myoepithelial cells also caused a significant decrease in MMP gene expression in co-cultured fibroblasts. Furthermore, this was associated with reduced gelatinolytic activity as identified by zymography. This study demonstrates for the first time that primary myoepithelial cells from normal breast reduce breast cancer cell invasion and that this is mediated via modulation of both tumour cell and fibroblast function. This emphasizes the importance of the myoepithelial cell in controlling the breast microenvironment and focuses on the potential significance of the loss of this population with disease progression.

Antineoplastic Agents↗
PubMed2003

LightCycler gyrA mutation assay (GAMA) identifies heterogeneity in GyrA in Salmonella enterica serotypes Typhi and Paratyphi A with decreased susceptibility to ciprofloxacin.

Mutations in gyrA in strains of Salmonella enterica serotypes Typhi and Paratyphi A have been characterised by a LightCycler-based PCR-hybridisation gyrA mutation assay (GAMA) and by DNA sequencing. Four mutations (Ser-83 to Phe, Asp-87 to Asn, Ser-83 to Tyr and Asp-87 to Gly) have been identified in 13 strains of Typhi and three strains of Paratyphi A resistant to nalidixic acid (=nal(r)) and with decreased susceptibility to ciprofloxacin (=Cp(L)), with the mutation Ser-83 to Phe predominating. The results have demonstrated heterogeneity in gyrA in nal(r) Cp(L) strains of Typhi and Paratyphi A and may be useful for epidemiological investigations. No mutations in gyrA were identified in four Cp(L) strains of S. Typhi that were sensitive to nalidixic acid. The mechanism of decreased susceptibility to ciprofloxacin in these strains is under investigation.

Anti-Bacterial Agents↗
PubMed2003

In vitro activity of piperacillin/tazobactam and other broad-spectrum antibiotics against bacteria from hospitalised patients in the British Isles.

Piperacillin/tazobactam is used for empirical therapy of severe and complex infections. We assessed its activity, 9 years after launch, against consecutive, clinically significant isolates from in-patients in UK and Ireland. Standardised disc susceptibility tests were performed on 5031 isolates at 28 hospitals. For quality assurance purposes, 5% of these isolates were collected centrally for MIC tests, as were those with exceptional resistances. Compared with a similar pre-launch survey in 1991, there were major increases in the proportions of Staphylococcus aureus, Pseudomonas aeruginosa, beta-haemolytic streptococci and Enterococcus faecium isolates collected, balanced by decreases in Escherichia coli, Proteus mirabilis and coagulase-negative staphylococci. These changes in species prevalence mostly favoured organisms with inherent resistance(s) or-in the case of S. aureus-reflected the massive increase of MRSA, up from 0.7% of all isolates in 1991 to 14.8% in 2001. Based on the disc tests, piperacillin/tazobactam retained activity against 87% of Enterobacteriaceae isolates, 95% of P. aeruginosa, 99% of streptococci and 96% of Enterococcus faecalis. Resistance nevertheless had increased since 1991 in E. coli from 4 to 10%, Klebsiella spp. (5 to 21%) and in AmpC-inducible Enterobacteriaceae (17 to 23%), though not in P. mirabilis or P. aeruginosa. MIC tests confirmed most of the piperacillin/tazobactam resistance found by disc tests in Enterobacter spp., but indicated susceptibility for about half of the E. coli isolates recorded as resistant in disc tests. This situation might be remedied by reducing the zone breakpoint, but this would increase the "false susceptible" rate unacceptably. Thus, if disc tests suggest that an isolate is marginally resistant to piperacillin/tazobactam and the drug is sought as therapy, it is recommended that MIC be determined with, e.g., an Etest.

Bacteria↗
PubMed2003

P-cadherin as a marker in the differential diagnosis of breast lesions.

AIMS: To assess the value of the calcium dependent cell adhesion molecule P-cadherin as a myoepithelial marker in the differential diagnosis of benign and malignant breast lesions. METHODS: Immunohistochemical analysis of normal breast, sclerotic breast lesions, tubular carcinomas, and ductal carcinoma in situ using a P-cadherin specific antibody and comparison with smooth muscle actin. RESULTS: All myoepithelial cells in normal breast ducts, ductules, and lobules and sclerotic lesions showed strong staining for P-cadherin. There was no staining of tubular carcinomas; myoepithelial cells were demonstrated around in situ carcinomas. Weaker reactivity was seen in a proportion of cells in some hyperplasias and in situ carcinomas. This weak reactivity in these tissues was not seen for smooth muscle actin but in radial scars, tubular carcinomas, and ductal carcinoma in situ staining of stromal cells caused difficulties in the identification of myoepithelial cells. CONCLUSION: P-cadherin is a useful marker in the differential diagnosis of breast lesions.

Adenocarcinoma↗
PubMed2003

There is more than one kind of myofibroblast: analysis of CD34 expression in benign, in situ, and invasive breast lesions.

AIMS: Smooth muscle actin (SMA) positive myofibroblasts have been implicated in tumour invasion; however, acquisition of SMA is not limited to peritumorous fibroblasts and other changes in fibroblasts may be more specifically related to the malignant environment. CD34 is a sialomucin expressed by normal breast fibroblasts but lost in invasive carcinomas. The aim of this study was to establish the relation between CD34 and SMA expression in breast fibroblasts and to analyse whether loss of CD34 is specific for invasive disease. METHODS: Immunohistochemistry for CD34 and SMA was performed on 135 cases including 10 normal, 10 fibroadenomas, 40 infiltrating ductal carcinomas, 55 cases of ductal carcinoma in situ (DCIS), and 20 radial scar/complex sclerosing lesions. The relation between staining pattern and histopathological features was recorded as positive, negative, or reduced. RESULTS: Fibroblasts around all normal duct-lobule units and those showing epithelial hyperplasia were CD34 positive and mainly SMA negative. In fibroadenomas, fibroblasts retained CD34 but acquired SMA expression. In contrast, fibroblasts around invasive carcinoma were CD34 positive and SMA negative. In DCIS, loss of CD34 was significantly more frequent in high grade tumours than in low or intermediate grade ones (p < 0.001). The acquisition of SMA was seen more frequently than the loss of CD34, particularly in non-high grade DCIS. In all radial scars, fibroblasts were SMA positive but CD34 negative, and a similar pattern was seen in stromal cells in areas of fibrosis following core biopsy. CONCLUSIONS: These results show that SMA positive myofibroblasts exhibit variable expression of CD34, indicating that these markers are not coordinately controlled. Loss of CD34 is strongly related to the malignant phenotype, in both invasive and preinvasive disease, but is not entirely specific because radial scar fibroblasts and fibroblasts in reactive fibrosis exhibit a similar phenotype. The functional relevance of altered CD34 expression is unclear but the very focal changes implicate local signalling mechanisms probably of epithelial origin.

Actins↗
PubMed2003

Expression of P-cadherin, but not E-cadherin or N-cadherin, relates to pathological and functional differentiation of breast carcinomas.

AIMS: To compare the expression of the cell adhesion molecules P-cadherin, N-cadherin, and E-cadherin in invasive and in situ breast carcinomas relative to clinicopathological features (size, node status, type, grade, and receptors) to determine whether expression patterns relate to specific tumour characteristics. METHODS: Using immunohistochemistry, 110 invasive and in situ breast carcinomas were examined for the presence, extent, and localisation of all three cadherins. Findings were related to tumour size, type, grade, node status, oestrogen (ER), progesterone, and epidermal growth factor receptor (EGFR) expression for invasive carcinomas and to grade and receptors for in situ carcinomas. RESULTS: P-cadherin was detected in 40% of invasive carcinomas, N-cadherin in 30%, and E-cadherin in 81%. For invasive carcinomas, the presence of P-cadherin significantly correlated with high grade, lack of ER and presence of EGFR, but not tumour size or node status. Carcinomas containing P-cadherin could be put into three categories dependent upon receptor and E-cadherin profile. There were no correlations between E/N-cadherin and size, grade, node status, or receptors. Three of 16 infiltrating lobular carcinomas expressed cytoplasmic but none membranous E-cadherin, and P-cadherin and N-cadherin were present in four carcinomas of this type. E-cadherin was found in all ductal carcinomas in situ, P-cadherin in a proportion of high grade tumours, and N-cadherin in a mixture of grades. CONCLUSION: P-cadherin but not E/N-cadherin expression in breast carcinomas shows a strong correlation with higher grade (poorer differentiation), lack of ERs, and presence of EGFR, and its expression may aid in the further subdivision of high grade carcinomas.

Biomarkers, Tumor↗
PubMed2002

The addition of glyceryltrinitrate to capsaicin cream reduces the thermal allodynia associated with the application of capsaicin alone in humans.

The aim of this study was to determine whether topical application of capsaicin cream causes thermal allodynia and the extent to which this is attenuated by the addition of glyceryltrinitrate (GTN). This was a double blind placebo controlled study of 40 consenting adult subjects. Each of four cream combinations (GTN, capsaicin, GTN/capsaicin and vehicle) were applied to the subjects with at least a 1 day interval between each application. Water at a known temperature was applied to the standard area of skin where cream had been applied. Subjects rated the resulting thermal allodynia using a 0-10 Likhert score. Thermal allodynia is usually apparent when warm water is applied to skin containing capsaicin. The thermal allodynia caused by the topical application of capsaicin was significantly reduced by the addition of GTN. The addition of GTN to capsaicin cream significantly reduces the thermal allodynia associated with the application of capsaicin cream alone.

Capsaicin↗
PubMed2002

The role of pedometers in the assessment of intermittent claudication.

OBJECTIVES: to evaluate the role of pedometers in the assessment of patients with intermittent claudication by comparing them with treadmill testing, post-exercise ankle brachial pressure index (ABPI) and physical function questionnaires. PATIENTS AND METHODS: fifty patients with intermittent claudication were prospectively assessed by treadmill examination measuring maximum walking distance (MWD) and post-exercise ABPI. Three physical function questionnaires: (1) Medical Outcome Study Short Form-36 (MOS SF-36), Physical Function (PF). (2) MOS SF-36 Role limitation due to Physical Problems (RP). (3) Walking Impairment Questionnaire (WIQ) were completed. Pedometers were used for one week and the daily average number of steps taken by each patient recorded. Pedometers scores were compared with MWD, post-exercise ABPI and physical function questionnaires scores using Spearman correlation test. RESULTS: pedometers estimation of daily physical activity moderately correlated with MWD (Rho=0.35, p=0.01), post-exercise ABPI (Rho=0.35, p=0.01), MOS-SF 36 PF (Rho=0.31, p=0.02), MOS-SF RP (Rho=0.28, p=0.05) and with WIQ (Rho=0.42, p=0.003). CONCLUSIONS: they moderately correlate with treadmill testing, post-exercise ABPI, and with physical function questionnaires. Pedometers may provide a simple and cheap alternative for the assessment of physical and functional disability in intermittent claudication.

Aged↗