Structural studies on heart muscle malate dehydrogenases.
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Transmission electron microscope observations on porcine enamel and secretory ameloblasts showed that silver methenamine material was located inside secretory vesicles in secretory ameloblasts and along the enamel crystals inside immature enamel. It is concluded that silver methenamine is able to stain enamel proteins selectively inside these tissues.
Proton-ligand dissociation and metal-ligand formation constants of azobenzene-N-malonic acid (I), p-Cl-azobenzene-N-malonic acid (II), p-Br-azobenzene-N-malonic acid (III) and p-COOH- azobenzene-N-malonic acid (IV) with UO2(2+), Th4+ and Ce3+ were evaluated potentiometrically using Bjerrum's method at 25, 35 and 45 +/- 0.5 degrees C and ionic strength 0.1 M in 40% v/v ethanol-water medium. The order of stability constants was found to be Ce3+ > Th4+ > UO2(2+). The effect of temperature on the dissociation and stability constants of the formed complexes was studied and the corresponding thermodynamic functions were derived and discussed. The ratio of metal-ligand was determined conductometrically. The structure of the ligands under investigation as well as their metal complexes has been elucidated by elemental analysis, IR and 1HNMR spectroscopy.
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Densitometric scans of electron micrographs of type I collagen segment long spacing crystallites stained with uranyl acetate or phosphotungstic acid and uranyl acetate have been correlated with computer-synthesized scans derived from the sequence of the alpha 1(I) and alpha 2 chains. Three models that differ in the location of the alpha 2 chain were used in the computer synthesis; Models A, B, and C have the alpha 2 chain in the A, B, and C chain positions, respectively. For all 13 experimental scans, the order of decreasing correlation with the models was found to be A,B,C. The probability of getting the same order of decreasing correlation all 13 times is 6/6(13). It was also determined at the 0.99 confidence level that the mean of the differences in the correlation coefficients among the models is greater than 0, supporting the conclusion that sequence-derived models best fit the experimental data when the alpha 2 chain is in the A position. Our results also agree with recent studies that show that uranyl ions bind to both positively and negatively charged residues on collagen type I.
Comparison of data from morphometric studies dealing with the heart is complicated by the fact that little information dealing with cell size changes during tissue processing is available. To investigate these changes, isolated cardiac myocytes were adhered to glass cover slips of Sykes Moore chambers and photographed after each step of processing for transmission electron microscopy. Six different experiments with a minimum of 10 cells each were followed through the entire procedure after fixation with isoosmolar glutaraldehyde. Cellular dimension changes were determined by tracing individual isolated myocytes after each step of the procedure with a sonic digitizer. Significant cell volume changes occurred after osmium (16 per cent swelling), postosmium wash (10 per cent swelling), and uranyl acetate (25 per cent shrinkage). Hypertonic aldehyde solutions resulted in cellular shrinkage during fixation not found with isotonic solutions. Changes in cell cross-sectional area rather than length were largely responsible for altered cell volumes during any given phase of processing. The results indicate that, although cell volume changes occur during processing, final cell dimensions of embedded cells were not different from unfixed cells. In whole tissue blocks, inclusion of propylene oxide in the procedure resulted in tissue shrinkage which was not observed in isolated myocytes, suggesting that different tissue components react in a variable manner to propylene oxide. After each of the other steps in processing, tissue blocks reacted in a similar manner to the isolated myocytes.
The modeling of geo-chemical processes needs the detailed and comprehensive knowledge of all chemical interactions existing in the flow path of waste dumps, subsoil and aquifer. This includes the adsorption, displacement and transport of heavy metal species of fulvic and humic acids, which represent the main amount of DOC in the liquid/solid system of the flow path. Comparative measurements of DOC concentrations in the input and output flow at the three waste dumps in the district Schlema/Alberoda indicated that DOC is produced and/or supplied within the waste dumps. The speciation of heavy metal compounds of fulvic and humic acids was achieved by means of sequential chromatographic analysis SCA and isoelectric focusing IEF. Between 5% and 20% of uranium exist in the flow path in the form of stable fulvic and humic acid species. Their distribution coefficients are strongly correlated with the pH values of the geo-chemical system.
It has recently been noted that a diversity of hyperthermophilic microorganisms have the ability to reduce Fe(III) with hydrogen as the electron donor, but the reduction of Fe(III) or other metals by these organisms has not been previously examined in detail. When Pyrobaculum islandicum was grown at 100 degrees C in a medium with hydrogen as the electron donor and Fe(III)-citrate as the electron acceptor, the increase in cell numbers of P. islandicum per mole of Fe(III) reduced was found to be ca. 10-fold higher than previously reported. Poorly crystalline Fe(III) oxide could also serve as the electron acceptor for growth on hydrogen. The stoichiometry of hydrogen uptake and Fe(III) oxide reduction was consistent with the oxidation of 1 mol of hydrogen resulting in the reduction of 2 mol of Fe(III). The poorly crystalline Fe(III) oxide was reduced to extracellular magnetite. P. islandicum could not effectively reduce the crystalline Fe(III) oxide minerals goethite and hematite. In addition to using hydrogen as an electron donor for Fe(III) reduction, P. islandicum grew via Fe(III) reduction in media in which peptone and yeast extract served as potential electron donors. The closely related species P. aerophilum grew via Fe(III) reduction in a similar complex medium. Cell suspensions of P. islandicum reduced the following metals with hydrogen as the electron donor: U(VI), Tc(VII), Cr(VI), Co(III), and Mn(IV). The reduction of these metals was dependent upon the presence of cells and hydrogen. The metalloids arsenate and selenate were not reduced. U(VI) was reduced to the insoluble U(IV) mineral uraninite, which was extracellular. Tc(VII) was reduced to insoluble Tc(IV) or Tc(V). Cr(VI) was reduced to the less toxic, less soluble Cr(III). Co(III) was reduced to Co(II). Mn(IV) was reduced to Mn(II) with the formation of manganese carbonate. These results demonstrate that biological reduction may contribute to the speciation of metals in hydrothermal environments and could account for such phenomena as magnetite accumulation and the formation of uranium deposits at ca. 100 degrees C. Reduction of toxic metals with hyperthermophilic microorganisms or their enzymes might be applied to the remediation of metal-contaminated waters or waste streams.
The geochemistry and microbiology of a uranium-contaminated subsurface environment that had undergone two seasons of acetate addition to stimulate microbial U(VI) reduction was examined. There were distinct horizontal and vertical geochemical gradients that could be attributed in large part to the manner in which acetate was distributed in the aquifer, with more reduction of Fe(III) and sulfate occurring at greater depths and closer to the point of acetate injection. Clone libraries of 16S rRNA genes derived from sediments and groundwater indicated an enrichment of sulfate-reducing bacteria in the order Desulfobacterales in sediment and groundwater samples. These samples were collected nearest the injection gallery where microbially reducible Fe(III) oxides were highly depleted, groundwater sulfate concentrations were low, and increases in acid volatile sulfide were observed in the sediment. Further down-gradient, metal-reducing conditions were present as indicated by intermediate Fe(II)/Fe(total) ratios, lower acid volatile sulfide values, and increased abundance of 16S rRNA gene sequences belonging to the dissimilatory Fe(III)- and U(VI)-reducing family Geobacteraceae. Maximal Fe(III) and U(VI) reduction correlated with maximal recovery of Geobacteraceae 16S rRNA gene sequences in both groundwater and sediment; however, the sites at which these maxima occurred were spatially separated within the aquifer. The substantial microbial and geochemical heterogeneity at this site demonstrates that attempts should be made to deliver acetate in a more uniform manner and that closely spaced sampling intervals, horizontally and vertically, in both sediment and groundwater are necessary in order to obtain a more in-depth understanding of microbial processes and the relative contribution of attached and planktonic populations to in situ uranium bioremediation.
Since the Gulf war exposure to depleted uranium, a known nephrotoxic agent, there is a renewed interest in the toxic effects of uranium in general and its mechanism of nephrotoxicity which is still largely unknown in particular. In order to investigate the mechanism responsible for uranium nephrotoxicity and the therapeutic effect of urine alkalization, we utilized rat renal brush border membrane vesicles (BBMV). Uranyl acetate (UA) caused a decrease in glucose transport in BBMV. The apparent K (i) of uranyl was 139+/-30 microg uranyl/mg protein of BBMV. Uranyl at 140 microg/mg protein of BBMV reduced the maximal capacity of the system to transport glucose [V (max) 2.2+/-0.2 and 0.96+/-0.16 nmol/mg protein for control and uranyl treated BBMV (P<0.001), respectively] with no effect on the apparent K (m) (1.54+/-0.33 and 1.54+/-0.51 mM for control, and uranyl treated BBMV, respectively). This reduction in V(max) is at least partially due to a decrease in the number of sodium-coupled glucose transporters as apparent from the reduction in phlorizin binding to the uranyl treated membranes, V (max) was reduced from 247+/-13 pmol/mg protein in control BBMV to 119+/-3 pmol/mg protein in treated vesicles (P<0.001). The pH of the medium has a profound effect on the toxicity of UA on sodium-coupled glucose transport in BBMV: higher toxicity at neutral pH (around pH 7.0), and practically no toxicity at alkaline pH (7.6). This is the first report showing a direct inhibitory dose and pH dependent effect of uranyl on the glucose transport system in isolated apical membrane from kidney cortex.
A series of uranyl aryloxide complexes has been prepared via metathesis reactions between [UO(2)Cl(2)(THF)(2)](2) and di-ortho-substituted phenoxides. Reaction of 4 equiv of KO-2,6-(t)()Bu(2)C(6)H(3) with [UO(2)Cl(2)(THF)(2)](2) in THF produces the dark red uranyl compound, UO(2)(O-2,6-(t)()Bu(2)C(6)H(3))(2)(THF)(2).THF, 1. Single-crystal X-ray diffraction analysis of 1 reveals a monomer in which the uranium is coordinated in a pseudooctahedral fashion by two apical oxo groups, two cis-aryloxides, and two THF ligands. A similar product is prepared by reaction of KO-2,6-Ph(2)C(6)H(3) with [UO(2)Cl(2)(THF)(2)](2) in THF. Single-crystal X-ray diffraction analysis of this compound reveals it to be the trans-monomer UO(2)(O-2,6-Ph(2)C(6)H(3))(2)(THF)(2), 2. Dimeric structures result from the reactions of [UO(2)Cl(2)(THF)(2)](2) with less sterically imposing aryloxide salts, KO-2,6-Cl(2)C(6)H(3) or KO-2,6-Me(2)C(6)H(3). Single-crystal X-ray diffraction analyses of [UO(2)(O-2,6-Cl(2)C(6)H(3))(2)(THF)(2)](2), 3, and [UO(2)Cl(O-2,6-Me(2)C(6)H(3))(THF)(2)](2), 4, reveal similar structures in which each U atom is coordinated by seven ligands in a pseudopentagonal bipyramidal fashion. Coordinated to each uranium are two apical oxo groups and five equatorial ligands (3, one terminal phenoxide, two bridging phenoxides, and two nonadjacent terminal THF ligands; 4, one terminal chloride, two bridging phenoxides, and two nonadjacent terminal THF ligands). Apparently, the phenoxide ligand steric features exert a greater influence on the solid-state structures than the electronic properties of the substituents. Emission spectroscopy has been utilized to investigate the molecularity and electronic structure of these compounds. For example, luminescence spectra taken at liquid nitrogen temperature allow for a determination of the dependence of the molecular aggregation of 3 on the molecular concentration. Electronic and vibrational spectroscopic measurements have been analyzed to examine trends in emission energies and stretching frequencies. However, comparison of the data for compounds 1-4 reveals that the innate electron-donating capacity of phenoxide ligands is only subtly manifest in either the electronic or vibrational energy distributions within these molecules.
A study was carried out of the resonance Raman scattering spectra of uranyl chloride (UO2Cl2) in dimethyl sulfoxide ((CH3)2SO) (DMSO) under laser excitation of the UO2(2+) ion in resonance with the 1sigma(g)+ --> 1phi(g) Laport-forbidden f-f electronic transitions span from 530 to 450 nm by using ten output lines of the argon-ion laser at room temperature. The resonance Raman excitation profile of the totally symmetric stretching vibrational mode of uranyl observed at 832 cm(-1) is presented and analyzed in terms of transform theory within the non-Condon model to give relatively good agreement with experimental results. The disagreement between the experimental data and the calculated resonance Raman excitation profile, at the long-wave part of the the 1sigma(g)+ --> 1phi(g) electronic transitions, may be referred to interference between the weak scattering from the neighboring forbidden electronic states (1delta(g)) and strong preresonance scattering from allowed electronic states at higher levels. An amount of change in the experimental resonance Raman excitation profile of the uranyl-DMSO system depends considerably upon the ligands (L) bound to the uranyl group. Elongation of the U-O equilibrium bond length resulting from the 1sigma(g)+ --> 1phi(g) electronic transitions is related to the magnitude of the change in the excitation profile of UO2L2 (L = NO3, CH3COO, Cl) type uranyl compounds in (DMSO).
The concentrations of several toxic heavy metals and volatile organic compounds (VOCs) in various types of Hungarian fly-ash fine particulates were investigated by means of instrumental neutron activation analysis, X-ray fluorescence analysis and gas chromatography, coupled with mass spectrometry. Within a power station, particulate samples were taken from the boiler zone (BO), from the electrostatic dust filter chamber (FI) and from the flue-gas at the top of the stack (ST). Enrichment rates of the toxic metals both in FI and ST particulate fractions related to the BO concentrations were calculated to enable the temperature dependence on the adsorption of the toxic components to be studied. In addition, both the total amounts of the VOCs and their partial distributions in accordance with the number of carbon atoms were also studied in fly-ash particulates. From them, 31 organic species were identified and determined. Since Hungarian brown coals have high uranium and thorium contents, the specific radioactivities of the daughter isotopes of both the 232Th and 238U decay series were also measured and are discussed.
The Geobacteraceae citrate synthase is phylogenetically distinct from those of other prokaryotes and is a key enzyme in the central metabolism of Geobacteraceae. Therefore, the potential for using levels of citrate synthase mRNA to estimate rates of Geobacter metabolism was evaluated in pure culture studies and in four different Geobacteraceae-dominated environments. Quantitative reverse transcription-PCR studies with mRNA extracted from cultures of Geobacter sulfurreducens grown in chemostats with Fe(III) as the electron acceptor or in batch with electrodes as the electron acceptor indicated that transcript levels of the citrate synthase gene, gltA, increased with increased rates of growth/Fe(III) reduction or current production, whereas the expression of the constitutively expressed housekeeping genes recA, rpoD, and proC remained relatively constant. Analysis of mRNA extracted from groundwater collected from a U(VI)-contaminated site undergoing in situ uranium bioremediation revealed a remarkable correspondence between acetate levels in the groundwater and levels of transcripts of gltA. The expression of gltA was also significantly greater in RNA extracted from groundwater beneath a highway runoff recharge pool that was exposed to calcium magnesium acetate in June, when acetate concentrations were high, than in October, when the levels had significantly decreased. It was also possible to detect gltA transcripts on current-harvesting anodes deployed in freshwater sediments. These results suggest that it is possible to monitor the in situ metabolic rate of Geobacteraceae by tracking the expression of the citrate synthase gene.
A modification of the tannic acid-metal salt method was applied as an ultrastructural stain for elastin. Thin sections of glutaraldehyde-fixed, embedded rat aorta and rabbit elastic cartilage, with and without osmication, were examined. Raising the pH of the tannic acid solution from 2.7 to 9.0 progressively increased the electron-density of elastic fibres and collagen fibrils in osmicated and unosmicated specimens. The maximum tannic acid staining of elastic fibres was observed in the pH range 7.0-9.0. Collagen staining, although less intense than that of elastic fibres, was also greatest in this pH range. Elastic fibres in osmicated specimens demonstrated the strongest tannic acid staining with a minimal increase in density of collagen and cell nuclei when compared to the unosmicated specimens. Sequential treatments of osmicated specimens with tannic acid pH 7.0-9.0, and uranyl acetate, pH 4.1, enhanced the density of the elastin intensely, increased collagen staining moderately, but hardly increased the density of nuclei and microfibrils. In elastase-digested osmicated specimens, all tannic acid (pH 7.0)-uranyl acetate-reactive elastin was selectively removed. These results demonstrate that all the neutral and alkaline tannic acid-uranyl acetate methods can be used as a postembedment stain for elastin specimens fixed in glutaraldehyde and osmium tetroxide.
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Responses in the frog glossopharyngeal nerve induced by electrical stimulation of the tongue were compared with those induced by chemical stimuli under various conditions. (a) Anodal stimulation induced much larger responses than cathodal stimulation, and anodal stimulation of the tongue adapted to 5 mM MgCl2 produced much larger responses than stimulation with the tongue adapted to 10 mM NaCl at equal current intensities, as chemical stimulation with MgCl2 produced much larger responses than stimulation with NaCl at equal concentration. (b) The enhansive and suppressive effects of 8-anilino-1-naphthalenesulfonate, NiCl2, and uranyl acetate on the responses to anodal current were similar to those on the responses to chemical stimulation. (c) Anodal stimulation of the tongue adapted to 50 mM CaCl2 resulted in a large response, whereas application of 1 M CaCl2 to the tongue adapted to 50 mM CaCl2 produced only a small response. This, together with theoretical considerations, suggested that the accumulation of salts on the tongue surface is not the cause of the generation of the response to anodal current. (d) Cathodal current suppressed the responses induced by 1 mM CaCl2, 0.3 M ethanol, and distilled water. (e) The addition of EGTA or Ca-channel blockers (CdCl2 and verapamil) to the perfusing solution of the lingual artery reversibly suppressed both the responses to chemical stimulus (NaCl) and to anodal current with 10 mM NaCl. (f) We assume from the results obtained that electrical current from the microvillus membrane of a taste cell to the synaptic area supplied by anodal stimulation or induced by chemical stimulation activates the voltage-dependent Ca channel at the synaptic area.
To monitor workers for external neutron radiation dose, the Y-12 National Security Complex utilises the thermoluminescence dosemeters (TLDs) manufactured by Harshaw. At Y-12, the majority of external dose to workers is due to low-energy photon and/or beta particles emitted from uranium and its progeny. However, some neutron dose is expected since neutrons are produced from (alpha,n) reactions in various compounds found at the plant, including UF4 and UF6. Neutron sources, such as 252Cf, are also used throughout the complex. The Harshaw neutron dosemeter consists of two gamma-sensitive elements (7Li) and two neutron-sensitive elements enriched in 6Li with various shielding/filter materials placed around each of them. In this work, the energy response of the dosemeter to neutrons has been calculated using the Monte Carlo transport code MCNP Version 4-C and, these results are compared with the measured response of the dosemeter to unmoderated and D2O-moderated 252Cf neutrons. The response of the dosemeter has also been determined in terms of the personal absorbed dose and personal dose equivalent as a function of neutron energy based on the recommendations of the ICRP Publication 60 and ICRU Report 49. The energy response of the dosemeter characteristics can be used to generate spectral conversion coefficients for routine neutron absorbed dose and dose equivalent calculations.