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Ion chromatographic determination of transition metals in irradiated nuclear reactor surveillance samples.

The determination of transition metal ions in radioactive (+/-25 microCi/g) low-alloy steels (nuclear reactor surveillance samples) by ion chromatography (IC) is described. The analysis has been done directly without prior separation of the iron matrix. The eluted metal ions have been detected with a UV-visible spectrophotometric detector after post-column complexation with 4-(2-pyridylazo)resorcinol. The results are in a good agreement with the certified values for the standard reference material used. The method was applied to nuclear reactor surveillance samples for the determination of Cu, Mn, Co and Ni.

Journal Article↗

Identification of the subunits of bovine heart mitochondrial NADH dehydrogenase that are exposed to the phospholipid bilayer by photo-labelling with 5-iodonaphth-1-yl azide.

Mitochondrial NADH dehydrogenase may be isolated from bovine heart as a lipoprotein complex (Complex I or NADH-ubiquinone oxidoreductase). Polypeptide subunits that are exposed to the hydrophobic region of the phospholipid bilayer were identified by photolabelling with the hydrophobic probe, 5-[125I]iodonaphth-1-yl azide. Chaotropic resolution of the labelled enzyme showed that the hydrophilic flavoprotein and iron-protein fragments of the enzyme were not in contact with the phospholipid bilayer. When complex I that had been partially depleted of phospholipids was photolabelled, incorporation of radioactivity into certain polypeptides was increased, indicating either conformational changes in protein or preferential association of these polypeptides with residual cardiolipin. A model NADH dehydrogenase structure is proposed on the basis of these results and those obtained with hydrophilic probes by Smith & Ragan (1980) Biochem. J. 185, 315-326.

Affinity Labels↗

Preservation of the pylorus in pancreaticoduodenectomy a follow-up evaluation.

We have previously reported our efforts to minimize postgastrectomy symptoms in two patients with benign disease who underwent resection of the head of the pancreas and the duodenum. In these cases the pylorus and first portion of the duodenum were preserved during pancreaticoduodenectomy. Our experience has now been extended to encompass 18 patients, eight of whom were available for comprehensive evaluation an average of six months postoperation. These studies have attempted to differentiate malabsorption of pancreatic insufficiency from possible gastrointestinal dysfunction of the new alimentary connection. Pancreatic insufficiency was evaluated by a 72-hour stool collection and radioactive trioctanoate (RATO) test. Gastrointestinal absorption was evaluated by D-xylose excretion and the Schilling test, as well as serum vitamin. A, vitamin B-12, carotene, folate, iron, and total iron binding capacity. Gastrointestinal secretion and motility were assessed by using pyloric fluoroscopy, gastric barium emptying, the Hunt test, and gastric acid analysis. Finally, a questionnaire regarding clinical symptoms of postgastrectomy syndromes and malabsorption was answered. Although every patient exhibited marked pancreatic insufficiency by laboratory tests, 88% described normal formed bowel movements, and weight loss was claimed by only 25%. Other test findings were generally normal. While the follow-up period has been limited to three years, the current data demonstrate that gastrointestinal function subsequent to preservation of the pylorus has not thus far predisposed to postgastrectomy syndromes or marginal ulcers. All of the patients required intensive pancreatic enzyme replacement.

Adolescent↗

The degradation of haem by carbon tetrachloride: metabolic activation requires a free axial coordination site on the haem iron and electron donation.

1. The ability of haem to catalyse the reductive activation of carbon tetrachloride (CCl4) in vitro has been investigated under anaerobic conditions, using methaemalbumin (MHA) and either sodium dithionite or NADPH together with NADPH-cytochrome P-450 reductase (EC 1.6.2.4) as the reducing agents. 2. In the non-enzymic system protohaem and other non-physiological haem analogues underwent rapid and extensive CCl4-dependent degradation, due to irreversible modification of their porphyrin tetrapyrrolic structure. 3. This mechanism of non-enzymic activation of CCl4 by protohaem mimics that catalyzed by cytochrome P-450 in that it requires a free, reduced haem iron and electron donation and it is largely prevented by carbon monoxide. 4. H.p.l.c. analysis of 14C-haem after anaerobic incubation with CCl4 and sodium dithionite gave radioactive products which eluted before and after haem, and exhibited significantly lower absorbance at 400 nm compared with authentic haem. When the products of CCl4-dependent haem degradation were methylated and applied to silica for t.l.c., two non-fluorescent pigments were isolated, purified and partially characterized. 5. On incubation of haem with 14CCl4 and sodium dithionite a 1:1 stoichiometry could be calculated for haem loss and 14CCl4-derived adduct formation, indicating that, as with microsomes, the loss of haem may be the result of a typical 'suicidal' inactivation reaction where the same haem moiety is both the site of CCl4 activation and the target of CCl4 reactive metabolites.

Animals↗

The binding of human lactoferrin to mouse peritoneal cells.

Human iron-saturated Lf (FeLf), which was labeled with 125I or 50Fe, was found to combine with the membrane of mouse peritoneal cells (MPC) which consisted of 70% macrophages. The following experimental data suggested the involvement of a specific receptor. (a) The binding of FeLf to MPC reached a saturation point. (b) The binding of radioactive FeLf was inhibited by preincubating the cells with cold FeLf but not with human Tf, human aggregated and nonaggregated IgG, or beef heart cytochrome c (c) Succinylation and carbamylation of FeLf resulted in a loss of its inhibiting activity on the binding of radioactive FeLf. Removal of neuraminic acid from FeLf increased its inhibitory activity. (d) The ability of apoLf to inhibit the binding of FeLf to MPC was significantly lower than that of FeLf. The existence of a Lf receptor capable of concentrating Lf released from neutrophils on the membrane of macrophages could explain the apparent blockade of the release of iron from the reticuloendothelial system, which accounts for the hyposideremia of inflammation. A receptor for FeLf was also found on mouse peritoneal lymphocytes. The affinity constant of FeLf for both lymphocytes and macrophages was 0.9 X 12(6) liter/mol. Howerver, macrophages bound three times more FeLf molecules (20 X 10(6)) per cell than did lymphocytes (7 X 10(6)).

Animals↗

Induction of an outer membrane protein of 78 kDa in Vibrio vulnificus cultured in the presence of desferrioxamine B under iron-limiting conditions.

Although Vibrio vulnificus is known to be able to utilize ferrioxamine B as an iron source, its outer membrane receptor remains to be determined. In this study, we found that V. vulnificus expressed a new outer membrane protein of 78 kDa when grown in the presence of desferrioxamine B under iron-limiting conditions. The desferrioxamine B-dependent iron uptake was only observed in bacterial cells expressing this protein. Furthermore, non-denaturing polyacrylamide gel electrophoresis followed by autoradiography of the outer membrane preparation containing the 78-kDa protein preincubated with [(55)Fe]ferrioxamine B provided a single radioactive band in which the 78-kDa outer membrane protein was present as the major component. These lines of evidence suggest that the inducible 78-kDa protein may serve as the cell-surface receptor for ferrioxamine B.

Amino Acid Sequence↗

In vitro assay of erythropoietin in fetal mouse liver cultures. I. Comparison of radioactive tracers and evidence of assay specificity.

In a 13 d fetal mouse liver tissue culture system for assay of erythropoietin, stimulation of incorporation of 3H-thymidine and of 125I-iodo-2'-deoxy uridine into DNA and of 59Fe iron into haem were compared as metameters of erythropoietin effect. With both DNA and haem tracers, log-dose response curves given by two preparations of human urinary erythopoietin purified to different extents (respectively 2 and 1300 IU, by bioassay in vivo, per mg of protein) were essentially identical at low dose levels although at high dose levels the less pure preparation decreased stimulation of both DNA and haem synthesis. Pre-incubation of erythropoietin with an antiserum to human erythropoietin abolished the effects of the hormones on both DNA and haem synthesis.

Animals↗

Biodynamic study of americium-241 accumulation in the cytosol of the hepatopancreas of the lobster Homarus gammarus.

In the lobster, most of the radionuclides ingested with contaminated food are concentrated in the digestive gland. Americium-241 accumulation in the hepatopancreas of the lobster was studied during the digestive cycle. Fractionations of cytosols at different times after ingestion of radioactive preys were performed by gel permeation chromatography to determine the distribution of 241Am in the different macromolecular components. 241Am was associated with ferritin during the whole digestive cycle. This observation suggests a correlation between 241Am distribution pathways and iron metabolism. The distribution of 241Am present in the other cytosolic proteins followed two major steps of accumulation which may be correlated to the evolution of the two main cellular types playing an important role in the digestive cycle (B and R type cells).

Americium↗

Modeling ligand-gated receptor activity. FhuA-mediated ferrichrome efflux from lipid vesicles triggered by phage T5.

An in vitro assay of iron-ferrichrome translocation across the FhuA protein of outer membranes from Escherichia coli has been devised. Upon reconstitution into large lipid vesicles, bacteriophage T5 binds to this polyvalent receptor, triggering a conformational change that resulted in channel opening. This facilitates the translocation of an iron(III)-siderophore, without the complexities involved in the in vivo process. Efflux of 55Fe(III)-ferrichrome across FhuA channels was determined quantitatively by monitoring the release of trapped radioactivity. The assay is rapid, reliable, and specific, because other bacteriophages, such as Phi80, fail to trigger channel opening of the FhuA receptor.

Bacterial Outer Membrane Proteins↗

Risk of lung cancer among iron ore miners: a proportional mortality study of 1,075 deceased miners in Lorraine, France.

A proportional mortality study of 1,075 iron ore miners in Lorraine, France, who died between 1960 and 1976 showed a significant excess of lung cancer mortality (proportionate mortality ratio = 2.25). Moreover, proportionate lung cancer mortality increased with the duration of work underground (proportionate mortality ratio = 4.24 for subjects who worked underground for more than 30 years) and was higher among pneumoconiotic (siderotic) miners (proportionate mortality ratio = 3.85) than among nonpneumoconiotic miners. These results were confirmed by a case-control study nested in the mortality study. Smoking habits could not be estimated retrospectively with sufficient accuracy to be taken into account. Although the proportion of smokers among contemporary iron ore miners is larger than in the French male population, occupational factors may also play a role; radiation exposure can be ruled out because there is no detectable radioactivity in the Lorraine mines, but dust exposure may be considered as an etiologic factor owing to the relationship between siderosis and lung cancer.

Adult↗

Uptake and conversion of the antibiotic albomycin by Escherichia coli K-12.

The antibiotic albomycin is transported into cells of Escherichia coli K-12 by the same uptake system as the iron-supplying ferrichrome complex. The iron-complexing hydroxamate moieties of albomycin and ferrichrome are structurally similar. During the phase of rapid iron uptake the chelators were not found in the cells. In order to understand the antibiotic activity of albomycin, it was labeled in the hydroxamate with tritium and in the presumed antibiotically active area with radioactive sulfur. While the tritium label was not retained by the cells, part of the sulfur label was taken up and concentrated 500-fold within the cell. The sulfur was not incorporated into proteins or nucleic acids since it was recovered as a low molecular weight component. Gel filtration on Bio-Gel P-2 revealed one tritium-labeled and two sulfur-labeled cleavage products in the incubation medium. We conclude that albomycin is actively transported via its ferrichrome-like portion into the cells and that the growth-inhibitory moiety is released by hydrolysis intracellularly and remains there.

Anti-Bacterial Agents↗

Riboflavin deficiency in the rat: effects on iron utilization and loss.

Iron absorption and daily loss of Fe were measured in riboflavin-deficient (B2-) Norwegian hooded rats and controls (B2+). Animals were fed on a test meal extrinsically labelled with 59Fe and whole-body radioactivity measured for 15 d. Riboflavin deficiency led to a reduction in the percentage of the 59Fe dose absorbed and an increased rate of 59Fe loss. All post-absorption 59Fe loss could be accounted for by faecal 59Fe, confirming that the loss was gastrointestinal. Fe concentrations and 59Fe as a percentage of retained whole-body 59Fe were higher in the small intestine of riboflavin-deficient animals than their controls, 14 d after the test meal. A separate experiment demonstrated that riboflavin deficiency was associated with a significant proliferative response of the duodenal crypts of the small intestine. These observations may explain the enhanced Fe loss in riboflavin deficiency.

Absorption↗

Relationship between tracheobronchial particle clearance rates and sites of initial deposition in man.

Mucociliary clearance was compared in three groups of normal subjects; each group inhaled a different type of aerosol (sebecate, iron oxide, and aqueous) labelled with 99Tc, to determine the relationship between deposition pattern and the subsequent total lung clearance. Standardization for deposition was accomplished by measuring a central to peripheral ratio (C/P) of deposited radioactivity in the thorax. Lung particle retention at 30, 60, 90, 120 min, and 24 h was measured, and compared at each time period to the C/P ratio. A linear relationship between the retention of aerosol at each time period and the C/P ratio was seen among all aerosols, e.g., aerosol with high C/P ratios had less retention of aerosol at a given time period than aerosols with more diffuse deposition patterns and lower C/P ratios. Furthermore, the relationship was similar to that of the more traditional measurement of regional deposition, the 24-h percentage retention. Lung particle retention was compared among aerosols, using analysis of covariance to standardize for C/P ratio. The intersubject variability was similar among all three aerosol groups. Thus, mucociliary clearance can be standardized in healthy subjects by using an initial deposition index, the C/P ratio, with similar accuracy to the 24-h percentage retention, as an index of aerosol deposition. The use of the C/P ratio, however, has distinct advantages over the 24-h retention, in that studies can be conveniently shortened, used with short lived isotopes, and be adapted to techniques investigating lung permeability instead of tracheobronchial clearance. An unexpected finding was the more rapid clearance of sebecate particles, compared with iron oxide or aqueous particles.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

241Am, 137Cs, Sr and Pb uptake by tobacco as influenced by application of Fe chelators to soil.

To determine the potential for phytoextraction of 241Am and other contaminants from soil, accumulation of 241Am, 137Cs, Sr, Fe, Al, Pb, and Mg by tobacco was determined for soil applications of two concentrations of ethylenediaminetetraacetic acid (EDTA), citric acid, and ascorbic acid. In tobacco receiving EDTA at 3.1 mmol/kg of soil, 241Am content of plants averaged 15 Bq/kg (ranging up to 26 Bq/kg) while Fe concentrations became constant at 4.5 mmol/kg. Soil treatment with 18.8 mmol/kg EDTA resulted in average 241Am concentrations of 29 Bq/kg (19 times higher than controls). Uptake of Pb was similar to 241Am. In these samples, Fe increased to a maximum of almost 18 mmol/kg and 241Am content increased linearly with both Fe and Al. Plants receiving ascorbic and citric acids took up smaller quantities of 241Am, Pb, and Fe, even though these reagents were able to elute about as much Fe from the soil as EDTA. Synchrotron microbeam X-ray fluorescence (SXRF) was used to determine radial distributions of elements in roots and stems with and without EDTA treatment. SXRF maps indicate differences in behavior between Fe and Pb that are consistent with the bulk plant observations and provide insight into changes in metal content of the roots in the presence of EDTA.

Americium↗

Behaviour of endocrine disrupting chemicals during the treatment of municipal sewage sludge.

Agricultural application of municipal sewage sludge has been emotionally discussed in the last decades, because the latter contains endocrine disrupting chemicals (EDCs) and other organic micropollutants with unknown fate and risk potential. Bisphenol A (BPA) was chosen as a model substance to investigate the influence of sludge conditioning on the end-concentration of EDCs in sludge. Adsorption studies with radioactive-labelled BPA showed that more than 75% BPA in anaerobically digested sludge is bound to solids (log Kd = 2.09-2.30; log Koc = 2.72-3.11). Sludge conditioning with polymer or iron (III) chloride alone had no influence on the adsorption of BPA. After conditioning with iron (III) chloride and calcium hydroxide desorption of BPA took place. Apparently, it occurred due to the deprotonation of BPA (pKa= 10.3) as the pH-value reached 12.4 during the process. The same behaviour is expected for other phenolic EDCs with similar pKa (nonylphenol, 17beta-estradiol, estron, estriol, 17alpha-ethinylestradiol). This study shows high affinity of BPA to the anaerobically digested sludge and importance of conditioning in the elimination of EDCs during the sludge treatment. Addition of polymer is favourable in the case of sludge incineration. Conditioning with iron (III) chloride and calcium hydroxide shows advantages for the use of sludge as fertiliser.

Absorption↗

Pharmacokinetics of superparamagnetic iron-oxide MR contrast agents in the rat.

The in vivo pharmacokinetics and the biodistribution of superparamagnetic iron-oxide particles (AMI25, Advanced Magnetics, Cambridge, MA) were investigated in anesthetized rats. Four different dose concentrations, ranging from 49.8 to 408.9 mumol of Fe (or 2.78-22.84 mg Fe) per kilogram, radiolabeled with 6.0 microCi of 59Fe-AMI25 were injected intravenously into 18 rats. The radioactivity cleared from the circulation with a fast component with a half-life of approximately 10 minutes and a slower component with a half-life of 92 minutes. Both half-lives were independent of the injected dose (ID) in the range of 105.4-408.9 mumol (5.89-22.84 mg) Fe/kg. The relative uptake in the liver, spleen, and kidneys was 57%, 2.9%, and 2.0% of the ID, respectively. At a dose of 52.1 mumol (2.91 mg) of Fe/kg, the relative concentration of iron significantly increased in the liver and decreased in the blood. Within the kidney, autoradiography showed that the iron was selectively taken up by the cortex. In the kidney, a concentration of 0.23 mumol (0.013 mg) Fe/g resulted in a 30% reduction in image intensity in a single echo magnetic resonance image obtained using a spin-echo sequence and an echo time of 70 ms.

Animals↗