The correlation between bismuth and uranyl staining and phosphorus content of intracellular structures as determined by electron spectroscopic imaging

Four groups of intracellular structures can be recognized according to bismuth and uranyl staining and phosphorus content. (1) Those which contain phosphorus and stain strongly with uranyl acetate but not with bismuth (ribosomes, heterochromatin and mature ribosomal precursor granules), presumably because of their nucleic acid content. (2) Those which contain phosphorus and stain with uranyl acetate and bismuth (interchromatin granules, immature ribosomal precursor granules and mitochondrial granules), presumably because at least some of their phosphate is available to react with bismuth. (3) Those which contain little phosphorus but which stain strongly with bismuth and weakly with uranyl acetate (Golgi complex beads), perhaps because some ligand in addition to phosphate reacts with bismuth, and (4) those which do not contain phosphorus and stain with neither uranyl acetate nor bismuth (portasomes). Uranyl staining correlates strongly with the phosphorus content of nucleic acids, proteins and inorganic deposits. Bismuth will stain some phosphorylated molecules but not all. Thus only some phosphates stain with bismuth.     

Toxicity of depleted uranium on isolated rat kidney mitochondria

Background

Kidney is known as the most sensitive target organ for depleted uranium (DU) toxicity in comparison to other organs. Although the oxidative stress and mitochondrial damage induced by DU has been well investigated, the precise mechanism of DU-induced nephrotoxicity has not been thoroughly recognized yet.

Methods

Kidney mitochondria were obtained using differential centrifugation from Wistar rats and mitochondrial toxicity endpoints were then determined in both in vivo and in vitro uranyl acetate (UA) exposure cases.

Results

Single injection of UA (0, 0.5, 1 and 2 mg/kg, i.p.) caused a significant increase in blood urea nitrogen and creatinine levels. Isolated mitochondria from the UA-treated rat kidney showed a marked elevation in oxidative stress accompanied by mitochondrial membrane potential (MMP) collapse as compared to control group. Incubation of isolated kidney mitochondria with UA (50, 100 and 200 μM) manifested that UA can disrupt the electron transfer chain at complex II and III that leads to induction of reactive oxygen species (ROS) formation, lipid peroxidation, and glutathione oxidation. Disturbances in oxidative phosphorylation were also demonstrated through decreased ATP concentration and ATP/ADP ratio in UA-treated mitochondria. In addition, UA induced a significant damage in mitochondrial outer membrane. Moreover, MMP collapse, mitochondrial swelling and cytochrome c release were observed following the UA treatment in isolated mitochondria.

General significance

Both our in vivo and in vitro results showed that UA-induced nephrotoxicity is linked to the impairment of electron transfer chain especially at complex II and III which leads to subsequent oxidative stress.     

Rapid incorporation of functional rhodopsin into nanoscale apolipoprotein bound bilayer (NABB) particles

Human apolipoprotein A-I (apo A-I) and its engineered constructs form discoidal lipid bilayers upon interaction with lipids in vitro. We now report the cloning, expression, and purification of apo A-I derived from zebrafish (Danio rerio), which combines with phospholipids to form similar discoidal bilayers and may prove to be superior to human apo A-I constructs for rapid reconstitution of seven-transmembrane helix receptors into nanoscale apolipoprotein bound bilayers (NABBs). We characterized NABBs by gel-filtration chromatography, native polyacrylamide gradient gel electrophoresis, UV-visible photobleaching difference spectroscopy, and fluorescence spectroscopy. We used electron microscopy to determine the stoichiometry and orientation of rhodopsin (rho)-containing NABBs prepared under various conditions and correlated stability and signaling efficiency of rho in NABBs with either one or two receptors. We discovered that the specific activity of G protein coupling for single rhos sequestered in individual NABBs was nearly identical with that of two rhos per NABB under conditions where stoichiometry and orientation could be inferred by electron microscopy imaging. Thermal stability of rho in NABBs was superior to that of rho in various commonly used detergents. We conclude that the NABB system using engineered zebrafish apo A-I is a native-like membrane mimetic system for G-protein-coupled receptors and discuss strategies for rapid incorporation of expressed membrane proteins into NABBs.     

On the chemiluminescence in the oxidation of tetravalent uranium to the uranyl ion by dimethyldioxirane

The reaction of the tetravalent uranium [U(IV)] with dimethyldioxirane (DMD) in strongly acidic water–acetone solutions is accompanied by chemiluminescence (CL) in the visible (Vis) and infra‐red (IR) regions. At least three independent reaction pathways are involved in the U(IV)–DMD oxidation: the first entails the non‐chemiluminescent oxidation of U(IV) to the uranyl ion (UO₂²⁺); the second involves the catalytic decomposition of DMD by U(IV) to afford singlet oxygen, as manifested by its characteristic IR‐CL; and in the third process, slow Vis‐CL (510–540 nm) is emitted, following DMD consumption. Copyright © 2002 John Wiley & Sons, Ltd.     

Purification and Characterization of Clathrin-Coated Vesicles from Chlamydomonas

Clathrin-coated vesicles, identified by negative staining with uranyl acetate, were purified from Chlamydomonas rein-hardtii. Isolated coated vesicles had diameters ranging from 70 to 140 nm (mean diameter±SD of 95±17 nm, n=300). These vesicles were markedly heterogeneous in both density and surface charge, as indicated by equilibrium density sedimentation and elution from anion-exchange columns. Highly-purified coated-vesicle fractions contained 2 major polypeptides, identified as the clathrin heavy chain (185 kDa) and the clathrin light chain (40 kDa). Chlamydomonas clathrin heavy chain cross-reacts weakly with an antibody against bovine brain clathrin heavy chain. Coat stability in several buffers was compared to that of bovine brain coated vesicles. Stability was similar, except for a greater stability of Chlamydomonas coated vesicles in 0.5 M Tris at pH 7.0.     

Synthesis and characterisation of a trinuclear uranyl complex: Crystal structure of (CN3H6)5[(UO2)3O(OH)2(CH3COO)(C2O4)3]

The reaction of uranyl oxalate trihydrate with guanidinium acetate at room temperature in water yields known uranyl complex with composition (CN₃H₆)₂[UO₂(C₂O₄)₂(H₂O)]·H₂O as a first phase and a novel complex (CN₃H₆)₅[(UO₂)₃O(OH)₂(CH₃COO)(C₂O₄)₃] as a second. The second phase was investigated by means of IR spectroscopy and X-ray diffraction. The trinuclear discrete complex contains two symmetrically independent uranyl ions with a pentagonal bipyramid structure and has a nonplanar geometry. The distortion of its equatorial plane is caused by substitution of a monodentate bridge hydroxide anion by a bidentate bridge acetate-anion. The acidic ligands found in the complex are usually in competition for a place in coordination sphere of an uranyl ion, thus peculiarities of the complex formation are discussed in terms of ‘crystallochemical analysis’.     

生物含磷大分子与铀酰相互作用模拟计算研究

【目的】针对UO₂²⁺_主要在生物细胞表面生成微纳米片状磷酸氢铀酰矿化体这一前期研究基础。【方法】本文采用模拟计算的方法,探究了磷脂（生物共有）、脂多糖（革兰氏阴性菌特有）、磷壁酸（革兰氏阳性菌特有）、植酸（植物特有）4种生物壁膜及胞外含磷大分子（biological macromolecules with phosphorous,BCP）与UO₂²⁺_之间的相互作用力、作用能和间距等。【结果】蒙特卡罗分析结果表明：在弱酸性(UO₂²⁺₎、中性至弱碱性(UO₂²⁺_与CO32–共存)两种体系下,BCP对UO₂²⁺_的吸附均容易发生,吸附能分布在–109 kcal/mol至–114 kcal/mol之间。分子动力学结果显示,UO₂²⁺     

The carbohydrates in frog retinal rod outer segments

Frog retinal rod outer segments appear to contain uncharacterized chemical components whose mass is roughly equivalent to 12--51% of the rhodopsin mass. Available data suggest that such components include soluble proteins and complex polysaccharides, and that hyaluronic acid accounts for a substantial fraction of this mass. Electron microscopic histochemical staining studies suggest that these polysaccharide components are located within the ROS disks. The oligosaccharide moieties of rhodopsin also appear localized within the disks. The interdisk cytoplasm may contain carbohydrates, but their quantity and identity are uncertain. Rhodopsin oligosaccharides as well as some fraction of the intradisk polysaccharide appear to have extended saccharide chains preferentially oriented perpendicular to the surface of the disk membrane. Possible roles for these polysaccharides in disk development and photoexcitation are discussed. The immediate need for complete rod outer segment chemical composition data is emphasized.     

Uranyl tris nitrato as a luminescent probe for trace water detection in acetonitrile

Uranyl tris nitrato i.e. [UO₂(NO₃)₃]^– was formed by adding tetramethylammonium nitrate to uranyl nitrate in acetonitrile medium. The luminescence features of this complex in acetonitrile are very sensitive to water content, which could lead to the use of it as a luminescent probe for water present in acetonitrile. The luminescence intensity ratio of 507 to 467 nm peak of uranyl tris nitrato showed a linear response in the range 0–5% (v/v) water content in acetonitrile. The present method was applied for three synthetic samples of acetonitrile for water detection and the results obtained were compared using Karl Fischer titration. There was a good agreement in the values obtained by both the methods.     

Effects of uranium poisoning on cultured preimplantation embryos

The toxic effect of uranium in cultured preimplantation embryos of the mouse is presented. Embryos were obtained from hybrid females CBA×C57 BL following induction of superovulation and were incubated in M16 cultured medium. Two different experiments were performed. In one, embryos in a one-cell stage were placed in culture media with final concentrations of uranyl nitrate of 104 and 208 μg/mL during 120 h in the same dish. In the other experiment, embryos in a one-cell stage were placed in culture medium with uranyl nitrate with final U concentrations of 26, 52, 104, and 208 μg/mL. At 24 h, those embryos which had reached the two-cell stage were transferred to another culture dish to which fresh solutions with uranyl nitrate were added. The percentage of embryos in two-cell stage, morula, early blastocyst, expanded blastocyst, and hatched blastocyst were recorded at 24, 72, 96 and 120 h of culture. The results obtained showed that concentrations as from 26 μg U/mL induced the delay of embryo development and the impairment of blastomere proliferation. The toxic effect of uranium increased in those experiments in which the embryos were transferred to a new medium. This embryo-culture system appears to be appropriate to evaluate the toxic effect of uranium on embryos removed from maternal influences and represents a suitable test system for environmental pollutants.