A comparison of three standard methods of identifying mast cells in endobronchial biopsies in normal and asthmatic subjects

Reported mast-cell counts in endobronchial biopsies from asthmatic subjects are conflicting, with different methodologies often being used. This study compared three standard methods of counting mast cells in endobronchial biopsies from asthmatic and normal subjects. Endobronchial biopsies were obtained from atopic asthmatic subjects (n=17), atopic nonasthmatic subjects (n=6), and nonatopic nonasthmatic control subjects (n=5). After overnight fixation in Carnoy's fixative, mast cells were stained by the short and long toluidine blue methods and antitryptase immunohistochemistry and were counted by light microscopy. Method comparison was made according to Bland & Altman. The limits of agreement were unacceptable for each of the comparisons, suggesting that the methods are not interchangeable. Coefficients of repeatability were excellent, and not different for the individual techniques. These results suggest that some of the reported differences in mast-cell numbers in endobronchial biopsies in asthma may be due to the staining method used, making direct comparisons between studies invalid. Agreement on a standard method is required for counting mast cells in bronchial biopsies, and we recommend the immunohistochemical method, since fixation is less critical and the resultant tissue sections facilitate clear, accurate, and rapid counts.

A nonradioactive method for the assay of polyphosphate kinase activity and its application in the study of polyphosphate metabolism in Burkholderia cepacia

Studies of polyphosphate (polyP) metabolism in microorganisms have been hampered by the lack of a convenient method for the assay in cell extracts of the activity of polyphosphate kinase (PPK), the enzyme principally responsible for microbial polyP biosynthesis. We report the development of such an assay, based on the well-established metachromatic reaction, with toluidine blue, of the polyP formed during the PPK-catalyzed reaction. The method was successfully used in the characterization of PPK activity in crude extracts of an environmental Burkholderia cepacia isolate. The development of a protocol for the physical recovery of polyP from solution is also reported. (C) 2002 Elsevier Science (USA). All rights reserved.

Enhancement of Ca2+-dependent outward current in sheep bladder myocytes by evans blue dye.

Whole-cell and inside-out patch-clamp techniques were used to assess the action of a well-known dye, Evans blue, on membrane currents in bladder isolated smooth muscle cells from sheep. In whole cells Evans blue dose-dependently increased the outward current by up to fivefold. In contrast, Evans blue had no effect on inward Ca2+ current. The effect on outward current was abolished or reduced if the cells were bathed in Ca2+-free solution, iberiotoxin (5 x 10(-8) M), or charybdotoxin (5 x 10(-8) M), but was unaffected by externally applied caffeine (5 mM) or in cells exposed to heparin (1 mg/ml) via the patch pipette. In inside-out patches bathed in a Ca2+ concentration of 5 x 10(-7) M, Evans blue (10(-4) M) increased the open probability of large-conductance (298-pS) Ca2+-dependent K+ channels (BK channels), shifting the half maximal-activation voltage by -70 mV. We conclude that Evans blue dye acts as an opener of BK channels.

A UKST survey of blue objects towards the Galactic centre - seven additional fields

We are searching for early-type stars towards the Galactic centre which are potentially young objects situated within the inner few kiloparsecs of the disk. Photographic photometry from the UK Schmidt Telescope has been used to identify the bluest candidates in nineteen Schmidt fields (centred close to the Galactic centre). We have previously obtained FLAIR low dispersion spectroscopy for three of these fields to estimate spectral types and here we present spectroscopy for an additional seven fields. Combining the results for all ten fields, 56 stars were initially classified as early-B type. Estimates of the equivalent widths of their Balmer and He I lines have been used to estimate atmospheric parameters and 32 targets have effective temperatures greater than or equal to 17 000 K (corresponding to a spectral type of B3 or earlier). The spectra of seven of these targets also have absorption lines due to O II and Si III and can be reliably classified as early- B type. Additionally 78 stars have estimated effective temperatures between 11 000 and 16 000 K with a further a further 50 objects identified as late-B (or early-A) type. All but two of the early B-type candidates have magnitudes in the range 12.0 less than or equal to V less than or equal to 16.0, and our best estimates of their distance suggest that they could be close to (i.e. R-g <3 kpc), or even beyond the Galactic centre.

Luminous blue variables as the progenitors of supernovae with quasi-periodic radio modulations

The interaction between supernova ejecta and circumstellar matter, arising from previous episodes of mass loss, provides us with a means of constraining the progenitors of supernovae. Radio observations of a number of supernovae show quasi-periodic deviations from a strict power-law decline at late times. Although several possibilities have been put forward to explain these modulations, no single explanation has proven to be entirely satisfactory. Here we suggest that Luminous blue variables undergoing S-Doradus type variations give rise to enhanced phases of mass loss that are imprinted on the immediate environment of the exploding star as a series of density enhancements. The variations in mass loss arise from changes in the ionization balance of Fe, the dominant ion that drives the wind. With this idea, we find that both the recurrence timescale of the variability and the amplitude of the modulations are in line with the observations. Our scenario thus provides a natural, single-star explanation for the observed behaviour that is, in fact, expected on theoretical grounds.

Between the devil and the deep blue sea: Nationality, power and symbolic trade-offs among evangelical Protestants in contemporary Northern Ireland

National identity is symbolically complex configuration, with shifts of emphasis and reprioritisations of content negotiated in contexts of power. This paper shows how they occur in one post-conflict situation - Northern Ireland - among some of the most extreme of national actors - evangelical Protestants. In-depth interviews reveal quite radical shifts in the content of their British identity and in their understanding of and relation to the Irish state, with implications for their future politics. The implications for understanding ethno-religious nationalism, nationality shifts and the future of Northern Ireland are drawn out.

Early birds are sexy: male age, dawn song and extrapair paternity in blue tits, Cyanistes (formerly Parus) caeruleus

Sexual selection theory predicts that signals reflecting the relative quality of individuals should be used in mate choice. Females could base their choice of copulation partners on male secondary sexual traits that honestly signal male age, as predicted by the age-based indicator mechanism. Studies have shown that female blue tits prefer older males and that aspects of dawn song reflect male quality, but it remains unknown whether dawn song characteristics correlate with male age. We compared dawn song characteristics of second-year (SY) and older (ASY) male blue tits (cross-sectional analysis), and tested for age-related changes within individuals (longitudinal analysis) and differential overwinter survival of SY males. We further investigated the relation between dawn song and paternity gain and loss. We found that ASY male blue tits began to sing earlier relative to sunrise than did SY males. This difference in the onset of dawn singing was due to age-related changes in individual performance rather than differential survival of individuals with varying expression of the trait. Males that began to sing earlier at dawn had more mating partners, and were more likely to gain extrapair paternity. Our findings suggest that the onset of dawn song can provide a simple mechanism for females to assess the relative quality of their mate and of neighbouring males. We propose that females use the onset of singing as a cue for their choice of extrapair partners. (c) 2006 The Association for the Study of Animal Behaviour. Published by Elsevier Ltd. All rights reserved.

Experimentally elevated plasma testosterone levels do not influence singing behaviour of male blue tits (Parus caeruleus) during the early breeding season

Sexual selection theory suggests that females might prefer males on the basis of testosterone (T)-dependent secondary sexual traits such as song. Correlational studies have linked high plasma T-levels to high diurnal song output. This has been confirmed in experiments where T-levels were kept high at times when natural T-levels have decreased. However, surprisingly little is known about the relation between T-levels during the early breeding season and song. In many passerine birds males sing at a high rate at dawn early in the breeding season, referred to as the dawn chorus. In blue tits (Parus caeruleus), the dawn chorus coincides with the fertile period of the female, whereas diurnal song occurs throughout the breeding season. Previous studies on blue tits showed that characteristics of the dawn chorus correlate with male reproductive success. We experimentally elevated plasma T-levels in male blue tits during the pre-fertile and fertile period. Our aim was to test whether increased plasma T-levels affect dawn song characteristics and increase the amount of diurnal song. Although T-implants successfully raised circulating T-levels, we did not find any difference between T- and control males in temporal performance measures of dawn song or in diurnal song output. Our results suggest that either there is no direct causal link between song output or quality and individual T-levels, or experimental manipulations of T-levels using implants do not permit detection of such effects during the early breeding season. Although we cannot exclude that individual T-levels are causally linked to other (e.g. structural) song parameters, our results cast doubt on T-dependence as the mechanisms that enforces honesty on song as a sexually selected trait.

The natural plasma testosterone profile of male blue tits during the breeding season and its relation to song output

In male birds, the gonadal hormone testosterone (T) is known to influence territorial and mating behaviour. Plasma levels of T show seasonal fluctuations which vary in relation to mating system and social instability. First, we determined the natural T profile of male blue tits Parus caeruleus during the breeding season. We found that plasma levels of T increased at the onset of nest building. Thus, the increase in circulating T was not associated with territory establishment, nor with the fertile period of the males' mates. In most individuals, T levels dropped to values close to zero during the period of chick feeding. Second, we investigated the relationship between plasma levels of T and male age, size, and singing behaviour. During the mating period, T levels did not differ between 1 yr old and older males and did not correlate with body size or condition. However, song output during the dawn chorus tended to be positively correlated with T levels. Therefore, if high T levels are costly, song output might be an honest indicator of male quality in blue tits. Finally, we show that plasma levels of T are significantly higher during the night than during the day. This pattern has also been observed in captive non-passerine birds, but its functional significance remains unknown.