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S E Bryan

Publications and source records attributed to S E Bryan.

At least 19 recordsLinked to original sources

Field and model investigations of external gamma dose rates along the Cumbrian coast, NW England.

A survey of the contribution to external dose from gamma rays originating from intertidal sediments in the vicinity of the British Nuclear Group Sellafield site showed that the major anthropogenic contributions were due to (137)Cs and (60)Co. At some sites, traces of other anthropogenic radionuclides were detected, namely (106)Ru, (125)Sb, and (154)Eu. The proportions of fine grained material (<63 microm) were used to improve model predictions of dose contribution due to external exposure to gamma rays, using the CUMBRIA77/DOSE77 model. Model dose predictions were compared to those directly measured in the field. Using the new proportions of fine grained material (1-17.5%) in conjunction with field gamma-ray spectra, model predictions were improved considerably for most sites. Exceptions were at Drigg Barn Scar and Whitehaven Coal Sands sites, which had their own unique characteristics. The highest (60)Co activity concentrations in this study were detected at Drigg Barn Scar. These relatively high activity concentrations of (60)Co were due to the presence of (60)Co in mussels and barnacles, hence upsetting the fine sediment relationships used in previous dose calculations. Whitehaven Coal Sands was unusual in that it contained higher levels of radionuclides than would be expected in sandy sediment. The mineralogy of these sediments was the controlling factor on (137)Cs binding, rather than the proportion of fine grained material. By adjusting the effective fine grained sediment proportions for calculations involving (60)Co and (137)Cs at Drigg Barn Scar and Whitehaven Coal Sands respectively, the CUMBRIA77/DOSE77 model predictions could be improved upon significantly for these sites. This work highlights the influence of particle size and sediment composition on external dose rate calculations, as well as the potential for external dose contributions from biota.

Animals↗

Comparison of measured and modelled copper binding by natural organic matter in freshwaters.

Fifteen freshwater samples containing significant concentrations of dissolved organic carbon-[DOC]-were titrated with copper under standardised conditions (pH 6 and 7), and concentrations of Cu(2+)-[Cu(2+)]-were measured with an ion-selective electrode. Measured values of [Cu(2+)], which were in the range 10(-11)-10(-5) moll(-1), were compared with those simulated using Humic Ion-Binding Models V and VI. It was assumed that copper speciation was controlled by the organic matter, represented by fulvic acid (FA), together with inorganic solution complexation (calculated with an inorganic speciation model). The models were calibrated by adjusting a single quantity, the concentration of FA. The optimised value-[FA](opt)-was that giving the best agreement, according to least squares, between measured and simulated [Cu(2+)]. The calculations took into account competition by other dissolved (filterable) metals (Mg, Al, Ca, Fe(II), Fe(III), Zn); in the case of Fe(III) it was assumed either that all the dissolved metal was truly in solution, or that the activity of Fe(3+) was controlled by equilibrium with Fe(OH)(3). The assumption about Fe(III) had relatively small effects on the fitting of Model V, but was significant for Model VI, because Model VI represents low-abundance, high-affinity binding sites in humic matter, which are sensitive to Fe(III) competition. Because of its inclusion of the high-affinity sites, Model VI provided better fits of the data than did Model V. Furthermore, Model VI with Fe(3+) activity controlled by Fe(OH)(3) gave smaller variation in the ratio of [FA](opt) to [DOC] than Model VI with all Fe(III) assumed to be in solution. The average [FA](opt)/[DOC] found from the Cu titrations was 1.30, which implies that 65% of the organic matter is 'active' with respect to metal binding. The average ratio of 1.30 is in reasonable agreement with ratios obtained by applying the model to field data sets for charge balance (1.22), Al speciation (1.56) and base titrations of Cu-amended waters (1.45). It is concluded that Model VI/Fe(OH)(3) provides the most reliable predictions of dissolved metal speciation in natural waters; at a total Cu concentration of 1 microM, the predicted concentration of Cu(2+) is expected to be correct to within a factor of 3.6 in 95% of cases.

Binding Sites↗

Infraspecific taxonomy and essential oil chemotypes in sweet basil, Ocimum basilicum.

Essential oil compositions of fresh and freeze-dried leaves were determined for 16 accessions of Ocimum basilicum belonging to different varieties to see whether they could be used as infraspecific taxonomic characters. One accession of O. x citriodorum was also studied. Some 30 monoterpenoids, sesquiterpenoids and phenylpropanoids were identified, the major components (more than 20% of the total essential oil composition in one or more accessions) being geranial and neral in O. x citriodorum, and linalool, methyl chavicol, eugenol, methyl eugenol and geraniol in O. basilicum. Based on a combination of the latter compounds, five major essential oil profiles could be distinguished in the accessions studied for O. basilicum. These profiles were largely the same for fresh and freeze-dried material of the same plant, although in the dried leaves, methyl chavicol and eugenol concentrations had generally declined in comparison to those of linalool. There appeared to be little correlation between essential oil patterns and varietal classification within O. basilicum. In view of the chemical heterogeneity of O. basilicum and its use as an essential oil-producing crop, culinary herb, medicinal plant and insect-controlling agent, in all of which chemicals play an important role, the infraspecific classification of this taxon should take chemical characters into consideration. A system for the classification of essential oil chemotypes in O. basilicum is proposed.

Gas Chromatography-Mass Spectrometry↗

Effects of ionic strength on endogenous nuclease activity in chelated and nonchelated chromatin.

Calf thymus chromatin, isolated using a standard (low ionic strength, but nonchelating) isolation protocol, dialyzed against either Tris-PMSF or Tris-EDTA, was reconstituted in a high salt compacting buffer (COM) or a low salt dispersing buffer (DIS) prior to digestion with endogenous nucleases. A greater level of enzyme activity occurred when chromatin was in a condensed state (COM buffer) and not chelated prior to digestion. In contrast, chromatin chelated by dialysis against Tris-EDTA prior to digestion showed higher levels of enzyme activity in the dispersed state (DIS buffer). Nonchelated undigested chromatin contained 0.280 +/- 0.16 ug copper/mg DNA and and 0.305 +/+- 0.09 ug zinc/mg DNA. Chelation removed about 78% of copper per mg DNA and approximately 65% of zinc per mg DNA. In COM buffer after a 20 min digestion, the solubilized fraction was enriched in copper showing about 20 X more metal per mg DNA than nonchelated chromatin. Approximately the same amount of zinc was found in both chelated and nonchelated chromatin while there was less zinc in chelated chromatin solubilized in DIS buffer. Thus, chelation has important effects on the digestibility of chromatin and on the type of ionic environment that provides the most favorable conditions for endogenous nuclease activity.

Animals↗

Angiotensin II receptors in chromatin fragments generated by micrococcal nuclease.

Rat liver nuclei were digested with micrococcal nuclease following incubation with 125I-angiotensin II (AII) or with 125I-AII and excess unlabeled hormone. Chromatin enriched in 125I was solubilized after 3 min and was applied to a BIO-GEL A-5 M column. Labeled hormone was 40-60% displaceable by unlabeled hormone, in nucleoprotein eluting with a V/Vo near 1.9, indicating that these solubilized chromatin fragments contained specific receptors for AII. Furthermore, a discrete AII binding nucleoprotein particle was resolved on DNP gel electrophoresis. Additionally, binding to specific AII nuclear receptors appeared to bring about changes in chromatin structure consistent with the induction of transcriptional activity.

Angiotensin II↗

Angiotensin II receptors in chromatin.

When isolated rat hepatic nuclei or bovine thymus nuclei were incubated with 125I-angiotensin II (ANG II) in the presence or absence of cold hormone, displaceable binding was consistently detected in micrococcal nuclease generated fragments Nanomolar concentrations of ANG II produced detectable displacement. Little or no specific binding was found when nuclei were first digested and then treated with hormone, suggesting that ANG II solubilizes its own receptor. The binding moiety was partially purified by DNP gel electrophoresis. These studies indicate the existence in chromatin of high affinity receptors for ANG II, and further suggest that hormone binding to these receptors produces conformational changes in chromatin similar to those seen during enhanced transcriptional activity. Thus, the present studies suggest the existence of functional intracellular renin-angiotensin systems.

Angiotensin II↗

Nuclear-hormone mediated changes in chromatin solubility.

Rat liver nuclei were incubated with either thyroid hormone or angiotensin (AII) at varying concentrations or with buffer (control) prior to digestion with micrococcal nuclease. Concentrations of hormones greater than 10(-10)M were effective in increasing the solubilization of chromatin with physiological levels (10(-9)M) of AII showing an approximate 2.4 fold increase over control. Nuclei were also isolated from animals treated in-vivo with either AII or buffer (control) and chromatin solubility was increased in the AII treated nuclei even prior to the addition of exogenous nuclease, presumably from the action of endogenous nucleases. The data suggest that hormone-induced increases in solubility are a reflection of structural changes in chromatin which enhance the accessibility of DNA to endonuclease attack.

Angiotensin II↗

Partitioning of zinc and copper within subnuclear nucleoprotein particles.

Nuclei from frozen calf thymus suspended in buffer were analyzed for metal content prior to and after repeated washing. After three such extractions about 0.1 micrograms Zn/mg DNA and 0.025 micrograms Cu/mg DNA remained tightly associated with chromatin. This level of metal was essentially unchanged with subsequent washings. Digestion of extracted nuclei with micrococcal nuclease yielded soluble nucleoprotein containing zinc and copper. Metal enriched regions of chromatin appeared to be preferentially solubilized by digestion, and the solubilized metal was only partially dializable either with or without EDTA. Metal profiles generated from gel (A-5m) chromatography analysis of chelated and non-chelated solubilized chromatin were distinctive in that copper was undetectable (by flame AA) while zinc was associated only with low molecular weight products when EDTA was used. In contrast, both metals were detected with higher molecular weight oligonucleosomes in the absence of chelating agents. Additionally, the two metals localized within nucleoprotein peaks and these metal-containing regions were only resolved by gel chromatography when EDTA was omitted throughout the procedure. A discrete Cu-rich species in a region of the profile suggests a subset of Cu-rich nucleoprotein complexes.

Animals↗

Cadmium, an effector in the synthesis of thionein.

Cadmium can elicit the synthesis of thionein in liver cells independent of tissue-organ interactions. The metal diffuses across the plasma membrane and is partitioned between subcellular components in a time dependent manner such that thionein synthesis responds to levels of nonspecifically and specifically bound cytoplasmic metal. Cadmium appears to function at the transcriptional level, and the metal may act to increase the pool of specific m-RNA's.

Animals↗

Induction of cadmium-thionein in isolated rat liver cells.

The uptake of cadmium by isolated liver cells was linearly related to the cadmium concentration to which the cells were exposed in the medium. Cadmium-treated cells synthesized proteins de novo with the characteristics of cadmium-thionein induced in the liver of cadmium-treated animals. Thionein from liver cells incorporated cadmium and [35S]cysteine, had a Ve/Vo (Sephadex G-50) of 1.8-1.9, and was separated into two subfractions by DEAE-cellulose ion-exchange chromatography. Cycloheximide and actinomycin D when added after a cadmium exposure prevented the synthesis of thionein. However, addition of actinomycin D after synthesis had started only decreased the total amount of thionein synthesized. The concentration of cadmium to which the cells were exposed affected the amount of cadmium-thionein synthesized in 6h. The maximum response occurred when cells were exposed to 0.5 microgram of cadmium/ml; at higher metal concentrations the total amount of cadmium-thionein synthesized declined. The system described in the present paper can be used to study the mode of metal toxicity and the mechanism of cadmium-thionein synthesis.

Animals↗

Interactions of mercury and copper with constitutive heterochromatin and euchromatin in vivo and in vitro.

Mouse liver nuclei were fractionated into (condensed) heterochromatin and (noncondensed) euchromatin by differential centrifugation of sonicated nuclei. The fractions were subsequently characterized as unique nuclear species by thermal denaturation derivative profile analysis, which revealed the heterochromatin fraction enriched in satellite DNA and by endogenous metal content, which displayed partitioning of mercury in euchromatin over heterochromatin by a 10:1 ratio, with a comparatively uniform distribution of copper in both fractions. Fractionation of nuclei following in vivo challenge with copper showed enrichment of copper in heterochromatin, relative to euchromatin, while in vivo exposure to mercury resulted in a 20-fold accumulation of mercury in euchromatin, relative to heterochromatin. Using gel filtration and equilibrium dialysis to measure in vitro binding under relatively physiologic conditions of pH (6.0-7.0) and ionic strength (standard saline citrate or saline), the condensed and noncondensed chromatin fractions exhibited binding specificities toward mercury and copper similar to that observed in the in vivo metal challenge experiments. The level of mercury which binds to euchromatin in vitro, when measured either in physiologic [standard saline citrate (SSC)] or in dilute (1:100 SSC) salt solutions, was comparable (approximately 3 mug of Hg/mg of DNA) to that of in vivo euchromatin-bound mercury after 1 month of challenge with dietary metal. In contrast, copper showed little or no preference for the nuclear fractions in dilute salt solutions and displayed patterns which mimic in vivo binding only at higher ionic strengths (saline). Removal of proteins from the chromatin fractions resulted in a loss of binding specificity toward both metals. Therefore, the binding selectivity of condensed and noncondensed chromatin toward both mercury and copper appears to arise from protein or from protein-DNA associations. The state of chromatin condensation is especially critical in the case of copper.

Animals↗