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Combined action of uranium and stress in the rat. I. Behavioral effects.

The influence of restraint stress on uranium (U)-induced behavioral effects was assessed in adult male rats. Eight groups of animals received uranyl acetate dihydrate (UAD) in the drinking water at doses of 0, 10, 20 and 40 mg/kg/day during 3 months. Rats in four groups were concurrently subjected to restraint during 2 h per day throughout the study. At the end of the period of uranium exposure, the following behavioral tests were carried out: open-field activity, passive avoidance and Morris water maze. Uranium concentrations in brain were also determined. At 10 and 20 mg/kg/day of UAD restraint significantly affected the total distance traveled in the open-field during the first and third periods tested, respectively, while no significant differences between groups were observed on the passive avoidance test. In the Morris water maze test, the influence of restraint was only significant on the latency time measured on Day 3 in rats exposed at 10 mg/kg/day. Restraint stress did not affect significantly the uranium levels in brain of rats. Although the results of the present study scarcely show uranium-induced behavioral effects at the oral doses of UAD here administered, these effects, as well as the slight influence of restraint stress noted in some tests should not be underrated.

Animals↗

Synthesis and reactivity of uranium(IV) amide complexes supported by a triamidotriazacyclononane ligand.

Reaction of [U{(SiMe2NPh)3-tacn}Cl] with LiNEt2 or LiNPh2 affords the corresponding amide compounds, [U{(SiMe2NPh)3-tacn}(NR2)] (R = Et (1), R = Ph (2)). The complexes have been fully characterized by spectroscopic methods and the solid-state structure of 1 was determined by single-crystal X-ray diffraction analysis. The six nitrogen atoms of the tris(dimethylsilylanilide)triazacyclononane ligand are in a trigonal prismatic configuration with the nitrogen atom of the diethylamide ligand capping one of the trigonal faces of the trigonal prism. Crystallization of 2 from CH3CN solution gave crystals of the six-membered heterocycle [U{(SiMe2NPh)3-tacn}{kappa2-(HNC(Me))2CC[triple bond]N}] (3). The reactivity of the amides was investigated. Both compounds undergo acid-base reactions with protic substrates such as HOC6H2-2,4,6-Me3, 3,5-Me2pzH (pz = pyrazolyl) and HSC5H4N to give the corresponding [U{(SiMe2NPh)3-tacn}X] (X = OC6H2-2,4,6-Me3 (4), 3,5-Me2pzH (5), kappa2-SC5H4N (6)) complexes. The solid-state structures of and were determined by single-crystal X-ray diffraction and revealed that the compounds are eight-coordinate with dodecahedral geometry.

Journal Article↗

[Bronchopulmonary precancerous conditions and tumors--risk groups from the occupational medicine viewpoint].

Risk groups with regard to bronchopulmonary precancerous and tumor diseases of occupational origin can be deduced from current occupational disease statistics. Most prominent are those working with asbestos. Each year about 250 asbestos-associated bronchial carcinomas and 400 mesotheliomas are recognized and compensated; the tendency is increasing. Because of the long latency time, the frequency peak will probably be reached in about 15 years in spite of the prohibition of asbestos usage. The second place is probably taken by malignomas among the underground uranium mine workers in Thuringia and Saxony (SDAG Wismut). Next come bronchial carcinomas with silicosis (carcinoma in scar tissue) after exposure to chromium(VI) and arsenic compounds as well as various other chemicals and metals. Dose-activity relationships are significant for all occupational carcinogenic agents, as there are also often syncancerogenic influences (especially smoking). From the data on previous loading, high risk groups, for example, among the insulation workers exposed to asbestos or uranium miners in the so-called "wild years", can be defined. A suitable screening method for the detection of bronchopulmonary tumors in the early stages has not yet been established. Medical checkups for the respective risk groups concentrate on the early X-ray detection of circular foci. As shown by recent studies, cytological sputum diagnosis, (fluorescence) bronchoscopy, and BAL cytology must be employed much more frequently in the high risk groups so that the prognostically more favorable stages of preneoplasm and carcinoma in situ can be detected and possibly treated curatively. These procedures are currently reaching a considerably higher sensitivity with the help of modern molecular biology techniques (e.g. detection of tumor-associated genetic changes and gene products). This contributes to an improvement in surveillance examinations with increasing detection of the curable early forms of tumors. However, only the further development of primary prevention, i.e. the greatest possible minimization or, if possible, total elimination of contact with carcinogenic agents and the consequent control of occupational protection will lead to a drastic reduction in the occupational risk of cancer.

Carcinoma, Bronchogenic↗

Partitioning and availability of uranium and nickel in contaminated riparian sediments.

The effects of iron oxides and organic matter on the partitioning and chemical lability of U and Ni were examined for contaminated riparian sediments from the U.S. Department of Energy's Savannah River Site. In sequential extractions of four sediments that ranged from 12.7 to 82.2 g kg(-1) in organic carbon, U was found almost exclusively in moderately labile fractions (93% in acid-soluble + organically bound). Nickel was distributed across all operationally defined fractions, including substantial amounts in the very labile fractions (4-15% in water-soluble + exchangeable), noncrystalline and crystalline iron oxides (38-49%), and in the nonlabile residual fraction (25-34%). Aqueous U concentrations in 1:1 sediment-water extracts were highly correlated to dissolved organic carbon (DOC) (R2 = 0.96; p < 0.0001) and ranged from 29 to 410 microg L(-1). Aqueous concentrations of Ni exceeded U by two to three orders of magnitude (124-2227 microg L(-1)) but were not correlated with DOC (R2 = 0.04; p = 0.53). Partitioning and solubility trends suggest that Ni availability is controlled primarily by iron-oxide phases, whereas U availability is dominated by naturally occurring organic carbon. Discrete mineral phases were also identified as nonlabile reservoirs of anthropogenic metals. In spite of comparably high sediment concentrations, Ni appears to be significantly more available than U in riparian sediments and therefore warrants greater consideration in terms of environmental consequences (i.e., transport, biological uptake, and toxicity).

Biological Availability↗

X-ray absorption spectroscopy studies of reactions of technetium, uranium and neptunium with mackinawite.

Technetium, uranium and neptunium may all occur in the environment in more than one oxidation state (IV or VII, IV or VI and IV or V respectively). The surface of mackinawite, the first-formed iron sulfide phase in anoxic conditions, can promote redox changes so a series of laboratory experiments were carried out to explore the interactions of Tc, U and Np with this mineral. The products of reaction were characterised using X-ray absorption spectroscopy. Technetium, added as TcO4(-), is reduced to oxidation state IV and forms a TcS(2)-like species. On oxidation of the mackinawite in air to form goethite, Tc remains in oxidation state IV but in an oxide, rather than a sulfide environment. At low concentrations, uranium forms uranyl surface complexes on oxidised regions of the mackinawite surface but at higher concentrations, the uranium promotes surface oxidation and forms a mixed oxidation state oxide phase. Neptunium is reduced to oxidation IV and forms a surface complex with surface sulfide ions. The remainder of the Np coordination sphere is filled with water molecules or hydroxide ions.

Environmental Monitoring↗

The effect of stress on the temporal and regional distribution of uranium in rat brain after acute uranyl acetate exposure.

Long-term exposure to depleted uranium (DU) has been shown to increase brain uranium and alter hippocampal function; however, little is known about the short-term kinetics of DU in the brain. To address this issue, temporal and regional distribution of brain uranium was investigated in male Sprague-Dawley rats treated with a single intraperitoneal injection of 1 mg uranium/kg as uranyl acetate. Due to the inherent stress of combat and the potential for stress to alter blood-brain barrier permeability, the impact of forced swim stress on brain uranium distribution was also examined in this model. Uranium in serum, hippocampus, striatum, cerebellum, and frontal cortex was quantified by inductively coupled plasma-mass spectrometry (ICP-MS) at 8 h, 24 h, 7 d, and 30 d after exposure. Uranium entered the brain rapidly and was initially concentrated in hippocampus and striatum. While multiple phases of uranium clearance were observed, overall clearance was relatively slow and the uranium content of hippocampus, cerebellum, and cortex remained elevated for more than 7 d after a single exposure. Prior exposure to stress significantly reduced hippocampal and cerebellar uranium 24 h post-exposure and tended to reduce uranium in all brain regions 7 d after exposure. The application of stress appeared to increase brain uranium clearance, as initial tissue levels were similar in stressed and unstressed rats.

Animals↗

Nephrotoxicity of simultaneous exposure to mercury and uranium in comparison to individual effects of these metals in rats.

Both inorganic mercury and uranium are known nephrotoxicants in mammals. In this study, the renal toxicity of a concurrent exposure to inorganic mercury and uranium was compared with the nephrotoxic effects of the individual metals in a rat model. Eight groups of rats, 10 animals per group, were subcutaneously given a single administration of mercuric chloride (HgCl2, 0.34 mg/kg and 0.68 mg/kg), uranyl acetate dihydrate (UAD, 2.5 mg/kg and 5 mg/kg), or combinations of both compounds at the same doses. A ninth group of rats received sc injections of 0.9% saline and was designated as the control group. Necrosis of proximal tubules, which was observed in all experimental groups, was the most relevant morphologic abnormality. Marked changes, which were remarkably greater than those induced by the individual elements, were noted in some urinary parameters in the groups concurrently exposed to HgCl2 and UAD. It could be an indicator of a synergistic interaction between mercury and uranium. In contrast, compared with the urinary levels found after individual administration of the highest doses of mercury and uranium, significant reductions in the urinary concentrations of these elements were noted following simultaneous exposure to both metals. At these doses, the reduction in the urinary metal excretion was also accompanied by significant decreases in the renal content of mercury and uranium. Whereas the results of some parameters pointed out a possible synergistic interaction between mercury and uranium, other measures hinted that an antagonistic interaction between these elements is also present.

Analysis of Variance↗

Comparison of early lung clearance of yellowcake aerosols in rats with in vitro dissolution and IR analysis.

The lung retention of uranium was determined in rats that inhaled aerosols of commercial yellowcake powders obtained from two mills (Mill A and Mill D) and whose chemical composition and solubilities in vitro were significantly different. Analysis by IR absorption indicated Mill A yellowcake contained 82% ammonium diuranate (ADU) and 18% U3O8. The Mill D powder contained 25% ADU and 75% U3O8. In vitro dissolution studies indicated for the Mill A sample, approximately 85% of the uranium had a dissolution half-time (T 1/2) of less than one day, with the remainder dissolving with a half-time of 500 days. For the Mill D sample, 25% had T 1/2 less than one day and 75% had T 1/2 of 300 days. Groups of 50 rats were exposed by nose-only inhalation to aerosols of either the Mill A or the Mill D yellowcake. Rats were sacrificed in groups of five at intervals through six months after exposure. Selected tissues and excreta samples were assayed by fluorometry to determine their uranium contents. For the Mill A yellowcake, 78% initial lung (broncho-alveolar) burden cleared with T 1/2 of 0.5 days, and 22% with T 1/2 of 240 days. For the Mill D yellowcake, 25% initial lung burden cleared with T 1/2 of 3.5 days and 75% with T 1/2 of 110 days. Thus, the lung clearance of uranium in the rat mimicked the in vitro dissolution data and supported the contention that ADU should be considered as a Class D compound (T 1/2 = 0.5 days) and U3O8 behaves in the lung as a Class Y (T 1/2 greater than 100 days) material.

Aerosols↗

Multiple influences of nitrate on uranium solubility during bioremediation of uranium-contaminated subsurface sediments.

Microbiological reduction of soluble U(VI) to insoluble U(IV) has been proposed as a remediation strategy for uranium-contaminated groundwater. Nitrate is a common co-contaminant with uranium. Nitrate inhibited U(VI) reduction in acetate-amended aquifer sediments collected from a uranium-contaminated site in New Mexico. Once nitrate was depleted, both U(VI) and Fe(III) were reduced concurrently. When nitrate was added to sediments in which U(VI) had been reduced, U(VI) reappeared in solution. Parallel studies with the dissimilatory Fe(III)-, U(VI)- and nitrate-reducing microorganism, Geobacter metallireducens, demonstrated that nitrate inhibited reduction of Fe(III) and U(VI) in cell suspensions of cells that had been grown with nitrate as the electron acceptor, but not in Fe(III)-grown cells. Suspensions of nitrate-grown G. metallireducens oxidized Fe(II) and U(IV) with nitrate as the electron acceptor. U(IV) oxidation was accelerated when Fe(II) was also added, presumably due to the Fe(III) being formed abiotically oxidizing U(IV). These studies demonstrate that although the presence of nitrate is not likely to be an impediment to the bioremediation of uranium contamination with microbial U(VI) reduction, it is necessary to reduce nitrate before U(VI) can be reduced. These results also suggest that anaerobic oxidation of U(IV) to U(VI) with nitrate serving as the electron acceptor may provide a novel strategy for solubilizing and extracting microbial U(IV) precipitates from the subsurface.

Ferric Compounds↗

Ultrastructural aspects of cytoplasmic ribosomes from Histoplasma capsulatum and Blastomyces dermatitidis as revealed by heavy metal staining.

Cytoplasmic ribosomes were isolated and purified from sonicates of the mycelial and yeastlike growth forms of the pathogenic dimorphic fungi, Histoplasma capsulatum and Blastomyces dermatitidis. Similar ribosomal fractions were prepared from Neurospora crassa and Saccharomyces cerevisiae. These latter organisms were selected as typical filamentous and yeastlike monophasic fungi, and their ribosomes were used as reference standards. High resolution electron microscopy permitted a comparison of both positively and negatively-stained ribosomes to those dehydrated without heavy metal salt. Such studies revealed statistically significant differences in physical dimensions. Cautious interpretations of substructural detail of the various ribosomal preparations suggested both interphasic and interspecies differences.

Blastomyces↗

Extraction of osmium-containing lipids by section staining for TEM.

Postfixation with osmium-ferrocyanide or OSO4 renders lipid droplets in rat liver and kidney homogeneously electron dense without additional section staining. In sections of the same block that have been single stained by uranyl acetate or lead citrate, lipid droplets show a more electron translucent center surrounded by a dense rim. In sections double stained with uranyl acetate and lead citrate, lipid droplets frequently appear as empty vacuoles, from which the electron dense content has been completely extracted.

Animals↗

The electron density of light and dark lysosomes in the proximal convoluted tubule of the rat kidney.

The present paper deals with the electron density of the lysosomal matrices in the renal proximal-convoluted-tubule cells of Wistar rats (20-60 days of age). Its purpose was to determine which physicochemical factors influence the electron density of lysosomes, and how their electron density is affected by various methods of fixation.--3 types of lysosomes can be distinguished in the proximal-convoluted-tubule cell; namely light, intermediate and dark lysosomes, which have an electron density lower, equal to or higher than the surrounding cytoplasm. All 3 types of lysosomes were invariably present after all methods of fixation tested. Light, intermediate and dark lysosomes differ in several respects. Staining intensity with uranyl acetate, lead citrate and potassium permanganate, osmiophilia, basophilia (in semithin sections) and --probably most importantly--the physical mass density of the lysosomal matrix are all low in light lysosomes, higher in intermediate lysosomes and highest in dark lysosomes. Light, intermediate and dark lysosomes of the proximal convoluted tubule do not form discrete classes, but one continuous spectrum of lysosomes of increasing electron density.

Animals↗

Mechanism of rhythmic contractions induced by uranyl ion in the ileal longitudinal muscle of guinea-pig.

The uranyl ion (UO2(2+)) produces rhythmic contractions of the longitudinal muscle of the ileum, similar to those induced by repetitive transmural stimulation. Hexamethonium inhibited the action of UO2(2+), indicating a preganglionic site of action of UO2(2+) and interneurons possibly being involved in the ACh-releasing effect of UO2(2+). In addition, the action of UO2(2+) was enhanced by physostigmine but antagonized by atropine, ATP, adrenaline and morphine suggesting multiple sites of action of UO2(2+). The effects of Ba2+ were studied simultaneously in order to compare them with those of UO2(2+). Atropine and hexamethonium had no effect on the rhythmic contractions with Ba2+ but physostigmine enhanced contractions induced by low concentration of Ba2+ indicating that Ba2+ induces contractions mainly by acting on the smooth muscle and releases ACh from the nerve to a lesser extent.

Acetylcholine↗

Examination of compact bone microdamage using back-scattered electron microscopy.

A new staining technique using heavy metals (lead-uranyl acetate) has been developed to allow visualization of bone microdamage using both light microscopy and scanning electron microscopy operated in its back-scattered mode (BSE). At the light microscopic level, the number of microcracks counted in sequential sections of human ribs is the same for both the traditional basic fuchsin method of differentially staining microcracks and the new lead-uranyl acetate procedure. With BSE study, however, the number of microcracks observed is significantly reduced in all samples, because of the reduction of projection effect error associated with surface based imaging techniques. Application of the lead-uranyl acetate staining technique to ex vivo-loaded crack propagation specimens showed an extensive ultrastructurally disrupted region associated with the crack path through bone, consistent with the damage process zone around cracks in toughened composite materials.

Adult↗

U(VI) adsorption to heterogeneous subsurface media: application of a surface complexation model.

The pH-dependent adsorption of U(VI) onto three heterogeneous, subsurface media from the Department of Energy (DOE) Oak Ridge Reservation, Savannah River Site, and Hanford Reservation was investigated. The three materials contained significant quantities of iron and manganese oxides with nearly identical extractable iron oxide contents (25.3-25.8 g/kg). A model independently developed for the adsorption of U(VI) to synthetic ferrihydrite (Waite, T. D.; Davis, J. A.; Payne, T. E.; Waychunas, G. A.; Xu, N. Geochim. Cosmochim. Acta 1994, 58, 5465-5478) was able to predict the major features of the pH-dependent U(VI) adsorption to the materials under the assumption that all the dithionite-citrate-bicarbonate extractable iron oxide was present as ferrihydrite. Further experiments with the Oak Ridge soil as a function of carbonate and U(VI) concentration indicated that the model could predict pH-dependent U(VI) adsorption to within a root mean square error of 0.163-0.408, even under conditions outside of those for which the model was developed. These results indicate that this model could be used as a first approximation in predicting U(VI) adsorption and transport in the subsurface. U(VI) adsorption also decreased at pH >10, even in the absence of carbonate, which is of potential importance to U(VI) mobility in extreme environments present in the subsurface at some DOE facilities. The pH-dependent adsorption of U(VI) was fundamentally different in systems with a constant CO2 partial pressure as compared to a constant total carbonate concentration. Experiments at constant CO2 partial pressure may not be representative of the conditions present in the subsurface, and a constant carbonate concentration does not always result in decreased U(VI) adsorption at higher pH values.

Adsorption↗

Luminescence quartz dating of lime mortars. A first research approach.

Lime mortars mixed with sand are well suited for connecting structural materials, like stones and bricks, due to the mechanical properties this material exhibits. Their extensive use in architectural and decorative works during the last 4000 years motivated the introduction of the 'Luminescence clock' for age determination of mortars. The same principles as for quartz optically stimulated luminescence (OSL) dating of sediments were applied for age estimation of a mortar fragment removed from a Byzantine church monument dated by archaeological means to 1050-1100 years ago (the first half of the 10th century). The OSL from the quartz was monitored under blue light stimulation and UV detection, using a single-aliquot-regenerative-dose protocol. The quartz-OSL dating of the mortar resulted in 870 +/- 230 a. TL polymineral fine grain dating was also performed on a brick fragment which was connected to the mortar, resulting in a TL age of 1095 +/- 190 a.

Calcium Compounds↗

Phosphorylation of vesicular stomatitis virus proteins as a possible contributing factor in virion uncoating.

The relationship between the in vitro phosphorylation of vesicular stomatitis virus (VSV) proteins and virion uncoating was examined. Activation of the VSV virion kinase with low concentrations of melittin, the active peptide component of bee venom, in the presence of gamma-[32P] ATP resulted in the phosphorylation of virion proteins. Following the in vitro phosphorylation of VSV proteins in the presence of melittin and deoxyadenosine triphosphate, the virion envelope was disrupted based on the accessibility of the internal ribonucleoprotein core (RNP) to the heavy metal stain, uranyl acetate, as determined by electron microscopic observation. The RNP structure was not observed in unphosphorylated virions treated with melittin and uranyl acetate. Phosphorylated virions treated with uranyl acetate subsequently lost the capacity for transcription whereas unphosphorylated virions treated with the stain retained transcriptase activity. These observations suggest that phosphorylation of VSV proteins may contribute to virion uncoating by disrupting the virus envelope.

DNA-Directed RNA Polymerases↗