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Induction of internucleosomal DNA fragmentation by carcinogenic chromate: relationship to DNA damage, genotoxicity, and inhibition of macromolecular synthesis.

Hexavalent chromium (Cr) compounds are respiratory carcinogens in humans and animals. Treatment of Chinese hamster ovary cells with 150 and 300 microM sodium chromate (Na2CrO4) for 2 hr decreased colony-forming efficiency by 46 and 92%, respectively. These treatments induced dose-dependent internucleosomal fragmentation of cellular DNA beyond 24 hr after chromate treatment. This fragmentation pattern is characteristic of apoptosis as a mechanism of cell death. These treatments also induced an immediate inhibition of macromolecular synthesis and delayed progression of cells through S-phase of the cell cycle. Cell growth (as evidenced by DNA synthesis) was inhibited for at least 4 days and transcription remained suppressed for at least 32 hr. Many of the cells that did progress to metaphase exhibited chromosome damage. Chromate caused the dose-dependent formation of DNA single-strand breaks and DNA-protein cross-links, but these were repaired 8 and 24 hr after removal of the treatment, respectively. In contrast, Cr-DNA adducts (up to 1/100 base-pairs) were extremely resistant to repair and were still detectable even 5 days after treatment. Compared with other regions of the genome, DNA-protein cross-links and Cr adducts were preferentially associated with the nuclear matrix DNA of treated cells, which was 4.5-fold enriched in actively transcribed genes. Chromium adducts, formed on DNA in vitro at a similar level to that detected in nuclear matrix DNA, arrested the progression of a DNA polymerase in a sequence-specific manner, possibly through the formation of DNA-DNA cross-links.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Chromium(III) induced abnormalities in human lymphocyte cell proliferation: evidence for apoptosis.

The occupational hazards and respiratory problems of workers associated with the chromium related industries like mining, electroplating and tanning have received much focus. Although the cytotoxic and mutagenic effects of common chromium(VI) compounds are now established, to the best of our knowledge, this is the first report of apoptosis (programmed cell death) caused by a set of Cr(III) complexes in human lymphocyte cells. The effects of five Cr(III) complexes with differing ligand environment and structure as well as K2Cr2O7 on Phytohaemagglutinin (PHA) induced lymphocyte cell proliferation have been investigated. Two of the Cr(III) complexes and K2Cr2O7 are found to cause apoptosis of lymphocytes but not the others. A case for the importance of species specific effects rather than non specific metal oxidation state dependent processes has now been made.

Apoptosis↗

Tannic acid and chromic chloride-induced binding of protein to red cells: a preliminary study of possible binding sites and reaction mechanisms.

The binding mechanisms and binding sites involved in the tannic acid and chromic chloride-induced binding of protein to red cells were investigated using the binding of IgA paraprotein to red cells as model systems. Inhibition studies of these model systems using amino acid homopolymers and compounds (common as red cell membrane constituents) suggest that the mechanisms involved are similar to those proposed for the conversion of hide or skin collagen to leather, as in commercial tanning. These studies also suggest that tannic acid-induced binding of IgA paraprotein to red cells involves the amino acid residues of L-arginine, L-lysine, L-histidine, and L-proline analogous to tanning with phenolic plant extracts. The amino acid residues of L-aspartate, L-glutamate and L-asparagine are involved in a similar manner in chronic chloride-induced binding of protein to red cells.

Binding Sites↗

[Immediate type allergies due to metal compounds: platinum salts].

Immediate type allergies due to metal compounds containing chromium, nickel, cobalt and platinum are rare. However, allergies to platinum salts represent a major health hazard in few precious metal refineries and catalyst productions. Sensitization can be shown with a highly specific and sensitive skin prick test. A dose-response relationship has clearly been established by prospective longitudinal studies, however the definition of a valid threshold limit is difficult due to the high variability of exposure. Smoking is a predicting factor, whereas atopy is not. Prognosis depends on the duration of symptomatic exposure in high exposure areas. Secondary prevention (medical surveillance) is effective in the prevention of persistent symptoms and bronchial hyperresponsiveness.

Humans↗

Evaluation of immunotoxic and immunodisruptive effects of inorganic arsenite on human monocytes/macrophages.

A trivalent inorganic arsenic, arsenite, has been causing chronic inflammation in humans through the consumption of contaminated well water. The total peripheral blood arsenic concentrations of chronic arsenic-exposed patients, who had inflammatory-like immune responses, are less than 1 microM, thus, nM concentrations may be very important regarding the chronic inflammatory effects by arsenite. However, there are few reports about the biological effects of low concentrations of arsenite in mammalian cells, especially in normal immune effector cells. In this study, we examined whether arsenite has any biological and/or toxicological effects on the differentiation of human peripheral blood monocytes into macrophages using the colony-stimulating factor (CSF) in vitro compared with that of other metallic compounds, and found that arsenite sensitively inhibited the CSF-induced in vitro maturation of monocytes into macrophages at nM levels, and it also induced small, nonadhesive and CD14-positive abnormal macrophage generation from monocytes with granulocyte-macrophage CSF (GM-CSF) at 50-500 nM without cell death. The addition of other metallic compounds, including chromium, selenium, mercury, cadmium, nickel, copper, zinc, cobalt, manganese and other human pentavalent arsenic metabolites, such as inorganic arsenate, monomethylarsonic acid and dimethylarsinic acid, could not induce the same abnormal cell generation from monocytes with CSFs at any concentration and any additional time schedules; they showed only simple cytolethality in monocytes and macrophages at n-mM levels accompanied by cell death. This work may have implications in the arsenic-induced chronic inflammation in humans.

Arsenates↗

Bond strengths of three chemical adhesive cements adhered to a nickel-chromium alloy for direct bonded retainers.

Sandblasted surfaces of a beryllium-free, nickel-chromium alloy were bonded with one of three chemical adhesives. After either immersion in water for up to 6 months or thermal cycling between 5 degrees and 60 degrees C for 500 cycles, the bonded specimens were tested for both shear and tensile strength. The highest values of tensile and shear bond strengths were found with Panavia Ex material, and these values showed no significant changes after thermal cycling. For ABC cement and for Super-Bond C & B material, the strength of the bond was significantly improved with thermal cycling. However, immersion in water for 6 months caused a significant decrease in the strength of the bond of specimens adhered with ABC cement.

Acrylates↗

Surface preparations for metal frameworks of composite resin veneered prostheses made with an adhesive opaque resin.

Bond strengths of a laboratory developed light-cured composite resin to dental casting alloys were evaluated with a new adhesive opaque resin. The metal specimens were type III gold, nickel-chromium, and cobalt-chromium alloys, while the surface treatments for bonding were heating, Sn plating, and ion coating. The cast metal specimens were "particle blasted" with aluminum oxide and were surface treated. Adhesive 4-META/MMA-TBB opaque resin was applied and a light-cured composite resin was placed over the opaque layer. The prepared specimens were thermocycled in water and shear bond strengths were recorded. The light-cured composite resin was bonded strongly to heated or Sn-plated type III alloy with 4-META/MMA-TBB opaque resin. Copper ion coating in a sputter coater was effective for all three alloys, with only slightly diminished bond strengths. These methods were satisfactory for making composite resin veneered prostheses.

Acrylic Resins↗

Spectroscopic approach in characterization of chromium(III), manganese(II), iron(III) and copper(II) complexes with a nitrogen donor tetradentate, 14-membered azamacrocyclic ligand.

The complexes of Cr(III), Mn(II), Fe(III) and Cu(II) were synthesized with the macrocyclic ligand i.e. 2,3,9,10-tetraketo-1,4,8,11-tetraazacyclotetradecane. The ligand was prepared by the [2 + 2] condensation reaction of diethyloxalate and 1,3-diamino propane. These complexes were found to have the general composition M(L)X3 and M'(L)X2 [where M = Mn(II) and Cu(II), M' = Cr(III) and Fe(III), L = ligand (N4) and X = Cl-, NO3-, 1/2SO4(2-) and [CH3COO-]. The ligand and its transition metal complexes were characterized by the elemental analyses, molar conductance, magnetic susceptibility, mass, IR, electronic, and EPR spectral studies. On the basis of IR, electronic and EPR spectral studies an octahedral geometry has been assigned for Cr(III), Mn(II) and Fe(III) and a tetragonal geometry for Cu(II) complexes.

Aza Compounds↗

Apoptosis of lymphocytes in the presence of Cr(V) complexes: role in Cr(VI)-induced toxicity.

Cr(VI) compounds have been declared as a potent occupational carcinogen by IARC (1990) through epidemiological studies among workers in chrome plating, stainless-steel, and pigment industries. Studies relating to the role of intermediate oxidation states such as Cr(V) and Cr(IV) in Cr(VI)-induced carcinogenicity are gaining importance. In this study, issues relating to toxicity elicited by Cr(V) have been addressed and comparisons made with those relating to Cr(VI) employing human peripheral blood lymphocytes. Lymphocytes have been isolated from heparinized blood by Ficoll-Hypaque density gradient centrifugation and exposed to Cr(V) complexes viz. sodium bis(2-ethyl-2-hydroxybutyrato)oxochromate(V), Na[Cr(V)O(ehba)(2)], 1 and sodium bis(2-hydroxy-2-methylbutyrato)oxochromate(V), Na[Cr(V)O(hmba)(2)], 2 and Cr(VI). The phytohemagglutinin (PHA)-induced proliferation of lymphocytes has been found to be inhibited by the two complexes of Cr(V) and chromate Cr(VI) in a time- and concentration-dependent manner. Viability of cells decreases in the presence of Cr(V). Apoptosis appears to be the mode of cell death in the presence of both Cr(V) and Cr(VI). Pretreatment of cells with antioxidants before exposure to chromium(V) complexes reverse apoptosis partially. Possibility for the formation and implication of reactive oxygen species in Cr(V)-induced apoptosis of human lymphocyte cells has been indicated in this investigation. The intermediates of Cr(V) and radical species in the biotoxic pathways elicited by Cr(VI) seems feasible.

Antioxidants↗

Shear bond strengths of three resin cements used with three adhesive primers for metal.

This study compared the durability and shear bond strengths of combinations of three adhesive primers and three resin cements bonded to silver-palladium-copper-gold (Ag-Pd-Cu-Au) and cobalt-chromium (Co-Cr) alloys. The adhesive luting cements Imperva Dual, Panavia 21, and Super-Bond C&B and the adhesive primers Metal Primer material, V-Primer material, and Cesead Opaque Primer material were used. The application of Metal Primer material was effective in improving the shear bond strengths between each of the three resin cements and Ag-Pd-Cu-Au alloy compared with nonprimed specimens. Co-Cr alloy primed with Cesead Opaque Primer, followed by cementation with Imperva Dual or Super-Bond C&B luting cements yielded the strongest shear bond strengths after 50,000 thermocycles, and Panavia 21 cement did not reveal any significant differences in bond strengths between nonprimed specimens and those primed with Cesead Opaque Primer at all thermocycles.

Adhesives↗

Evaluation of inert markers for the determination of ileal and faecal apparent digestibility values in the pig.

A series of experiments was designed to evaluate inert markers employed in studies of ileal and faecal apparent digestibilities of nitrogen and amino acids in pigs fitted with simple 'T' piece cannulas. Trial 1 assessed the palatability of diets containing (a) 5 g chromic oxide/kg, (b) 1 and (c) 5 g titanium dioxide/kg at two levels of feeding. The results indicated that there were slight problems in terms of period of feed consumption associated with diets (a) and (c), but that these effects were transitory and disappeared following acclimatization, although higher levels of feeding may compound the problem. Trial 2 estimated N balance, faecal apparent digestibility of amino acids as determined by both total collection and ratio of markers, ileal apparent digestibility of amino acids and recovery of markers using diets containing (a) 1 and (b) 5 g Cr2O3/kg, (c) 1 and (d) 5 g TiO2/kg. An additional assessment of acid-insoluble lignin present within the diet was also undertaken. Cr2O3 was associated with the lowest recovery, whilst calculations based on TiO2 gave lower standard errors. Trial 3 examined diurnal variation in digesta sampling and its effect on faecal apparent digestibility of N and amino acids. There were no evident effects. In general, it was concluded that the most appropriate marker to use in studies of this nature was TiO2 at a rate of 1 g/kg.

Amino Acids↗

APFIM investigations on site occupancies of the ternary alloying elements Cr, Fe, and Re in NiAl.

The site occupancies of the transition metals Cr, Fe, and Re dissolved in NiAl of stoichiometric composition have been determined by atomic layer resolved atom probe field-ion microscopy (APFIM). The investigations were supported by X-ray diffraction studies to evaluate the lattice parameters. These are influenced by atomic size effects and constitutional lattice defects like Ni antistructure atoms in the Al sublattice and vacancies in the Ni sublattice. The APFIM results were compared with ALCHEMI data and calculated site preference energies published in the literature. Chromium additions to stoichiometric NiAl with 0.8 at% in solid solution exhibit a strong preference for Al sites. The lattice parameter of NiAl(Cr) solid solution is decreased. Iron atoms dissolved in higher concentrations of 5 at % in NiAl are almost equally distributed within both sublattices. They are possessing a weak preference for Al sites, which causes a lattice expansion of NiAl(Fe) solid solution. ALCHEMI results and site preference energy data show a strong site preference of Cr atoms for the Al sublattice. In contrast, iron atoms exhibit a weak site preference for Ni sites depending upon the stoichiometry of the NiAl host lattice. Re solutes in low concentrations of about 0.2 at % in NiAl possess a strong site preference for the Ni sublattice. The increase of the lattice parameter of NiAl(Re) is due to the pronounced size effect of Re atoms. For these species no ALCHEMI and site preference energy data are available in the literature.

Alloys↗

Adhesive bonding of stainless steels and their component metals.

The purpose of the current study was to evaluate the bond strength of a metal adhesive system bonded to stainless steels and their component metals. Two sizes of disk specimens (10 and 8 mm in diameter x 2.5 mm thickness) were machined from two stainless steels designed for magnetic attachment (AUM20 and SUS 316L), as well as from high-purity chromium (Cr) and nickel (Ni) metals for reference. The specimens were air-abraded with alumina, either primed with a metal conditioner (Cesead II Opaque Primer) or left unprimed, and bonded with an adhesive resin (Super-Bond Opaque). Shear bond strengths were determined before and after thermocycling, and the results were analysed by analysis of variance (ANOVA). Post-thermocycling bond strengths of the unprimed groups were 16.3 MPa for the AUM20 alloy, 7.5 MPa for the SUS 316L alloy, 31.1 MPa for Cr and 3.1 MPa for Ni. Those of the conditioned groups were 30.3 MPa for the AUM20 alloy, 32.9 MPa for the SUS 316L alloy, 39.3 MPa for Cr and 13.1 MPa for Ni. Application of the conditioner elevated the bond strengths of all groups (P<0.05). It can be concluded that combined use of the conditioner and the Super-Bond adhesive is effective for bonding the stainless steels examined, and that Cr is a suitable component for the bonding system in question.

Analysis of Variance↗

Reduction of Cr(VI) at a polyaniline film: influence of film thickness and oxidation state.

Hexavalent chromium [Cr(VI)] is highly toxic, carcinogenic, and mutagenic to living organisms. In this paper, the reduction of Cr(VI) to the much less toxic trivalent state [Cr(III)] was studied at polyaniline films grown to different thickness. Much higher rates of Cr(VI) reduction were observed for the "thick" polyaniline films. This was explained in terms of the morphology of the polymer and the higher surface area of polymer in contact with the Cr(VI) solution. For "thin" polyaniline films, the Cr(VI) reduction reaction was found to obey pseudo-first-order kinetics for the duration of exposure. However, in the case of thick polyaniline layers, the Cr(VI) reduction reaction followed a two-stage process, with each stage obeying pseudo-first-order kinetics. This was explained in terms of oxidation of the polymer from the leucoemeraldine to the emeraldine state and then further oxidation of the polymer from the emeraldine to the pernigraniline state. Much higher rates of Cr(VI) reduction were observed on oxidation of the polymer from the leucoemeraldine to the emeraldine state.

Aniline Compounds↗

The DNA repair inhibitors hydroxyurea and cytosine arabinoside enhance the sensitivity of the alkaline single-cell gel electrophoresis ('comet') assay in metabolically-competent MCL-5 cells.

We have found previously that the metabolically-competent human MCL-5 cell line did not appear to be usefully sensitive to the DNA-damaging effects of several carcinogens, as measured by the alkaline single-cell gel electrophoresis ('comet') assay. We therefore sought to increase its sensitivity by inhibiting DNA repair during exposure to test compounds, using 10 mM hydroxyurea (HU) and 1.8 mM cytosine arabinoside (ara-C), which inhibit DNA resynthesis during nucleotide excision repair. The following compounds were tested, using a 30-min exposure, in the absence or presence of HU/ara-C: 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (8-MeIQx), 2-amino-3,4,8-trimethylimidazo[4,5-f]quinoxaline (4, 8-DiMeIQx), 2-amino-3-methylimidazo[4,5-f]quinoline (IQ), 2-amino-9H-pyrido[2,3-b]indole (A[alpha]C), 2-amino-3-methyl-9H-pyrido[2,3-b]indole (MeA[alpha]C), 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), benzo[a]pyrene (B[a]P), 3-methylcholanthrene (3-MCA), 7, 12-dimethylbenz[a]anthracene (DMBA), 1-nitropyrene (1-NP), 2-nitrofluorene (2-NF), aniline, o-toluidine, benzene, lindane, bleomycin, cisplatin, N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), sodium chromate, chromic chloride, and diethylstilboestrol (DES). We made the following observations. The background level of comet formation was reasonably constant over several months and was increased only slightly, but significantly, in the presence of the DNA-repair inhibitors. All compounds that induced comet formation did so without appreciable cytotoxicity as assessed by trypan blue exclusion. Of the compounds tested, the heterocyclic amines and polycyclic aromatic hydrocarbons (with the exceptions of PhIP and B[a]P) failed to induce convincing levels of comet formation in the absence of repair inhibitors. In their presence the heterocyclic amines tested induced comet formation (with the exception of 8-MeIQx), with widely differing potencies. 1-NP failed to elicit marked comet formation even in the presence of HU/ara-C. Aniline and o-toluidine produced significant levels of comet formation in the absence of HU/ara-C, but in their presence comet formation was markedly increased. Benzene, lindane, bleomycin, cisplatin, MNNG, sodium chromate and chromic chloride induced comet formation in the absence of HU/ara-C, but, with the exception of cisplatin, their presence enhanced comet formation. Neither sucrose nor DES elicited comet formation under the conditions used in this study. Many more agents need to be tested in order to determine how well the comet assay using MCL-5 cells (or modified versions of it) can distinguish genotoxins from non-genotoxins.

Amines↗

Bond strength of composite to alloy treated with bonding systems.

PURPOSE: The in-vitro bond strengths of a composite bonded to a nickel-chromium-beryllium alloy treated by eight bonding systems were measured after three storage conditions. MATERIALS AND METHODS: Nickel-chromium-beryllium alloy samples were treated by eight commercial bonding systems including adhesive composite cements, all-purpose bonding agents, and silica-coating systems. A composite was bonded to the alloy samples. Sample groups were stored in water for 24 hours at 23 degrees C, or thermocycled, or stored for 6 months at 23 degrees C and then debonded in tension. RESULTS: Bond strengths after 24 hours storage were: adhesive composite cements, 14.2 to 22.1 MPa; all-purpose bonding agents, 11.4 to 14.6 MPa; and silica-coating systems, 18.6 to 20.2 MPa. Bond strengths after thermocycling were: adhesive composite cements, 12.6 to 20.6 MPa; all-purpose bonding agents, 9.9 to 17.7 MPa; and silica-coating systems, 11.1 to 19.2 MPa. Bond strengths after 6 months were: adhesive composite cements, 12.0 to 13.1 MPa; all-purpose bonding agents, 8.7 to 14.1 MPa; and silica-coating systems, 14.8 to 18.4 MPa. CONCLUSIONS: Only two bonding systems showed decreased bond strength after thermocycling, as compared with 24 hours' storage; whereas four bond systems showed decreased bond strength after 6 months' storage. Bond strengths measured after thermocycling were not predictive of those measured after 6 months' storage.

Analysis of Variance↗

Chromium-mediated synthesis of polycyclic aromatic compounds from halobiaryls.

[reaction: see text] Reaction of 2,2'-dihalobiphenyl with butyllithium followed by the addition of chromium(III) chloride and alkynes afforded the corresponding phenanthrene derivatives via formal [4 + 2] cycloaddition. A variety of alkynes could be used for this reaction, such as alkyl, aryl, silyl, and alkoxycarbonyl alkynes. Repetitive process of the reaction gave more extended polycyclic compounds such as benzo[g]chrysene and azacyclopentaphenalene derivatives.

Journal Article↗

Comparison of bond strengths of three denture base resins to treated nickel-chromium-beryllium alloy.

PURPOSE: In-vitro bond strengths of 3 denture base resins (Trutone, Lucitone 199, and Triad) to a nickel-chromium-beryllium removable partial denture alloy (Ticonium) were tested with 3 surface pretreatments: sandblast, acid etch, and Rocatec (silica blasting), with or without primers (Dentsply, CR inlay cement, and Super Bond). MATERIAL AND METHODS: Lucitone 199 denture base resin bonded to the nonprimed sandblasted alloy specimen served as the control group. Alloy specimens were prepared, surface treated, and primed or not primed with primer. The treated specimens were then packed and processed with the denture base resin. Bonded specimens were stored in the distilled water at 37 degrees C for 24 hours, and then debonded in tension. The force at which the bond failed was noted, and bond strength was calculated in megapascals (MPa). Five replications for each condition (180 specimens total) were tested. RESULTS: Significant differences in bond strength were observed with primer, the most important factor, followed by pretreatment and denture base resin. Without primer for all 3 denture base resins, the Met-Etch and Rocatec treated group showed significantly higher bond strengths than the sandblasted groups. For Trutone denture base resin, nonprimed treated groups produced significantly higher bond strength than those for the other 2 denture base resin, nonprimed treated groups produced significantly higher bond strength than those for the other 2 denture base resins. The control group had zero bond strength. For Dentsply primer, the Rocatec treated group bonded to Lucitone 199 resin produced the highest bond strength value (14.8 +/- 1.8 MPa). For CR inlay cement, the Met-Etch and Rocatec treated groups for Lucitone denture base resin demonstrated the highest bond strength (19.3 +/- 4.8 MPa, and 19.3 +/- 1.8 MPa, respectively). For super Bond primer, the Met-Etch treated group for Trutone resin demonstrated the highest bond strength (19.8 +/- 6.2 MPa). CONCLUSIONS: Without primer, the control had the lowest bond strength (0 MPa), and the Trutone groups showed the highest bond strength (11.7 +/- 4.1 MPa). Met-Etch and Rocatec treated groups produced higher bond strengths than the sandblasted groups. The primed specimens demonstrated significantly higher bond strengths than nonprimed specimens, except for Trutone resin, for which primed specimens produced lower bond strengths than the nonprimed specimens.

Acid Etching, Dental↗