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Formation of the paramagnetic complex [Cr(OH)5O2]5- in the reaction between chromium(VI) oxide, and hydrogen peroxide or superoxide anion radicals studied by EPR spectroscopy.

Hydrogen peroxide or superoxide anion radicals form a paramagnetic complex in the reaction with chromium(VI) oxide in an alkaline water solution at room temperature. The complex [Cr(OH)5O2]5- with the g-value equal to 1.9734 is believed to contain hydroxyl groups derived from the alkaline solution and dioxygen derived from hydrogen peroxide or superoxide anion radicals.

Chromium Compounds↗

Effect of chromium on apolipoprotein A-I expression in HepG2 cells.

OBJECTIVE: Chromium is a key micronutrient required for lipid and carbohydrate metabolism. Some but not all clinical trials have associated use of chromium supplements with improved insulin sensitivity and lipid profile including increased high-density lipoprotein cholesterol levels. METHODS: Because apolipoprotein A-I (apoA-I) is the principal protein of high-density lipoprotein, the molecular pathways underlying chromium-related changes in apoA-I expression were studied in a human hepatoma cell line (HepG2) transfected with full-length apoA-I promoter attached to the reporter chloramphenicol acetyl transferase gene. RESULTS: Exposure of these cells to different concentrations of chromium chloride (0, 0.5, 1.0, and 3.0 mM) resulted in a dose-dependent decrease in apoA-I promoter activity (chloramphenicol acetyl transferase activity expressed as a percentage of an internal control was 99.4 +/- 7.2% in control cells versus 87.6 +/- 5.0%, 73.4 +/- 2.3%, and 36.6 +/- 3.9%, respectively, P < 0.01). Chromium chloride at 10 mM concentration was toxic and caused death in a large number of cells. Treating HepG2 cells with other minerals known to have insulin-sensitizing effects such as magnesium (1 mM), zinc (0.2 mM), and vanadyl sulfate (0.1 mM) significantly reduced apoA-I promoter activity in the presence and absence of 100 microU/mL of insulin. Northern blot analyses showed that the apoA-I mRNA content of cells treated with 0.2 mM of chromium chloride relative to G3PDH mRNA was not significantly increased compared with controls (0.652 +/- 0.122 versus 0.745 +/- 0.143, the ratio of apoA-I to glyceraldehyde 3-phosphate dehydrogenase (G3PDH) mRNA in control and chromium-treated cells, respectively). Western blot analyses of proteins secreted in culture media indicated that neither chromium treatment of the HepG2 cells (858.0 +/- 151.4 arbitrary units) nor treatment with magnesium (1323.3 +/- 175.7) or vanadium (1102 +/- 78.7) significantly altered apoA-I concentrations compared with controls (1061.7 +/- 114.7). However treatment of HepG2 cells with 0.2 mM of zinc significantly reduced apoA-I concentrations (291.0 +/- 29.2 versus 1061.7 +/- 114.7; P < 0.001). CONCLUSIONS: Supraphysiologic concentrations of chromium and other minerals with known insulin-sensitizing activity may reduce apoA-I promoter activity in cultured cells. Whether similar changes may occur in vivo remains to be shown. However, these observations do not support the use of pharmacologic amounts of chromium supplementation to enhance the cardioprotective lipid profile.

Antioxidants↗

Synthesis of chromium-containing pigments from chromium recovered from leather waste.

Cobalt-chromite green and chrome-tin pink pigments have been prepared from chromium extracted from leather shavings produced as a waste product of the leather-tanning industry. The alkaline agent (NaOH, CaO, MgO, NH4OH) used in the extraction process influences the nature of the final product. The effect of the NH4OH:CaO ratio on the final product in the case of cobalt-chromite green was examined. The pigments obtained were characterized by FT-IR and X-ray diffraction (XRD). Colour measurements were compared with those recorded from materials prepared from pure Cr2O3. The leaching of Cr(VI) from the materials was examined by UV spectroscopy.

Chromium↗

Systematic investigations of the contrast results of histochemical stainings of neurons and glial cells in the human brain by means of image analysis.

The investigation of neurohistological specimens by image analysis has become an important tool in morphological neuroscience. The problems which arise during the processing of these images are non-trivial, especially if a pattern recognition of cells in the imaged tissue is intended. One of the major problems faced concerns the segmentation of structures of interest, whether cells or other histologic structures. The segmentation problem is often the result of an inappropriate staining procedure. For serious image analysis to be performed, the material under investigation must be optimally prepared. Spatially complex patterns, e.g. fuzzy-like neighbouring neurons, are easy to recognize for humans. But the integrative and associative performance of current artificial neuronal network schemes is too low to achieve the same recognition quality as humans do. Therefore, a general analysis of staining characteristics was performed, especially with respect to those stains which are relevant to object segmentation. Although most image analytical investigations of tissues are based on stained samples, a study of this type has not been previously conducted. Of the stains and procedures evaluated, the gallocyanin chrome alum combination staining provided the best stain contrast. Furthermore, this staining method shows sufficient constancy within different parts of the human brain. Even the fine nuclear textures are differentiable and can be used for further pattern recognition procedures.

Adult↗

Use of colloids of chromic [32P] phosphate in treatment of solid tumors.

In order to evaluate the effectiveness of an intratumorally single dose of chromic [32P] phosphate for the treatment of solid tumors, studies of bioelimination, biodistribution, and therapeutic action were carried out. Only for comparative purposes were similar studies undertaken using a solution of sodium [32P] orthophosphate-gelatin. Results show that when sodium [32P] orthophosphate-gelatin was intratumorally injected, the percentage of total elimination, after 32 days of treatment, was equal to 85.90 +/- 8.70%, with a higher percentage in urine (64.50 +/- 13.70%) than in feces (21.40 +/- 4.50%). In biodistribution studies, the greater percentage was found in bone (15.54 +/- 2.21%), whereas only 2.51 +/- 0.39% remained in the tumor. When chromic [32P] phosphate was intratumorally injected, we found that the total elimination was equal to 51.70 +/- 6.90%, with a higher amount in feces (32.70 +/- 4.80%) than in urine (19.00 +/- 3.60%). Biodistribution studies demonstrated that 28.93 +/- 1.30% was still in the tumor and 19.01 +/- 1.30% of the injected activity was found in the liver. On the other hand, when therapeutic action was evaluated, no tumoral regression was observed. These results demonstrate that the colloid of chromic [32P] phosphate cannot be used in the treatment of solid tumors as it mobilizes from the injection point, delivering a high dose to the entire organism.

Adenocarcinoma↗

Gastrointestinal transit times in young and old cats.

Ageing results in a decrease in apparent nutrient digestibility in the gastrointestinal (GI) tract. The aim of this study was to investigate whether the rate of gastric emptying or total GI transit times differed between young (3.0+/-0.9 years) and senior (11.6+/-1. 4 years) cats. Gastric emptying rates were measured using [1-(13)C]octanoic acid and total transit times with chromium oxide. No significant differences (P>0.05) were observed in either the rate of gastric emptying or total transit time between young and senior cats although senior cats exhibited a larger variability in total transit time compared to the younger cats (35.71+/-14.06 and 26. 46+/-5.80 h, respectively). The results of this study indicate that the observed reduction in nutrient digestibility in ageing cats is not due to alterations in the rate of passage of digesta through the GI tract.

Aging↗

A comparison of the in vitro genotoxicity of tri- and hexavalent chromium.

Chromium can be found in the environment in two main valence states: hexavalent (Cr(VI)) and trivalent (Cr(III)). Cr(VI) salts are well known human carcinogens, but the results from in vitro studies are often conflicting. Cr(VI) primarily enters the cells and undergoes metabolic reduction; however, the ultimate product of this reduction, Cr(III) predominates within the cell. In the present work, we compared the effects of tri- and hexavalent chromium on the DNA damage and repair in human lymphocytes using the alkaline single cell gel electrophoresis (comet assay). Potassium dichromate induced DNA damage in the lymphocytes, measured as the increase in comet tail moment. The effect was dose-dependent. Treated cells were able to recover within a 120-min incubation. Cr(III) caused greater DNA migration than Cr(VI). The lymphocytes did not show measurable DNA repair. Vitamin C at 50 microM reduced the extent of DNA migration. This was either due to a decrease in DNA strand breaks and/or alkali labile sites induced by Cr(VI) or to the formation of DNA crosslinks by Cr(VI) in the presence of vitamin C. Vitamin C, however, did not modify the effects of Cr(III). Catalase, an enzyme inactivating hydrogen peroxide, decreased the extent of DNA damage induced by Cr(VI) but not the one induced by Cr(III). Lymphocytes exposed to Cr(VI) and treated with endonuclease III, which recognizes oxidized pyrimidines, displayed greater extent of DNA damage than those not treated with the enzyme. Such an effect was not observed when Cr(III) was tested. The results obtained suggest that reactive oxygen species and hydrogen peroxide may be involved in the formation of DNA lesions by hexavalent chromium. The comet assay did not indicate the involvement of oxidative mechanisms in the DNA-damaging activity of trivalent chromium and we speculate that its binding to cellular ligands may play a role in its genotoxicity.

Carcinogens, Environmental↗

Effect of chlorophyllin on chromium trioxide-induced micronuclei in polychromatic erythrocytes in mouse peripheral blood.

The effect of chlorophyllin on micronucleated polychromatic erythrocytes (MN-PCE) induction by chromium trioxide (CrO(3)) exposure in peripheral blood of mice was studied. Animals were treated with a single intraperitoneal dose of chlorophyllin (CHL) (20mg/kg), CrO(3) (20mg/kg), and CHL (20mg/kg) 4h before (CHL-CrO(3)) or 4h before and 20h after chromium treatments (20mg/kg; CHL-CrO(3)-CHL). Peripheral blood samples were drawn from the caudal vein at 0, 12 and 48h, and analyzed by the acridine orange (AO) technique. The results obtained in present study shown that CHL injection did not modify the number of MN-PCE. CrO(3) treatment resulted in a significantly increases 12 and 48h after the injection, reaching a four-fold increase 48h after CrO(3) administration. Whereas treatment with 20mg/kg of CHL prior to chromium, decreased the MN frequency induced by chromium in the 12h samples. When the samples were analyzed 48h after CrO(3) injection, no significant differences between CHL-CrO(3) and CHL-CrO(3)-CHL in comparison with CrO(3) treatment, were observed. These results indicate that increase of MN-PCE by CrO(3) is CHL-blocked in both protocols used (CHL-CrO(3) and CHL-CrO(3)-CHL) at 12h after treatment, but it was unable to modify the frequency of MN-PCE measured 48h after CrO(3) injection. The absence of a protective effect by CHL in the MN-PCE induction by CrO(3) at 48h, show that CHL has action only on one of the times of MN induction and suggests the possible action of CrO(3) by two different mechanisms, and not by CHL time-limited in vivo.

Animals↗

Evidence suggesting that chlorophyllin (CHLN) may act as an inhibitor or a promoter of genetic damage induced by chromium(VI) oxide (CrO3) in somatic cells of Drosophila.

In Drosophila, 48h-old larvae were pretreated for 24h with chlorophyllin (CHLN) or sucrose and then treated with chromium(VI) oxide (CrO(3)) immediately following completion of the pretreatment period (0-day delay) or delayed 1, 2 or 3 days. The effects were scored in the wing spot test. After delays of 0 and 1 day, clear evidence of a protective effect of CHLN was found. Contrarily, after delays of 2 and 3 days, the results showed a reversal, i.e. CHLN-related events appeared more frequently than those in the sucrose control suggesting a promoting effect. It would appear prudent that CHLN be tested in a variety of situations in any given organism before decisions are reached regarding its inhibitor/promoter effects.

Animals↗

Detection of genotoxic effects of heavy metal contaminated soils with plant bioassays.

Aim of the present study was the development of a bioassay which enables the detection of genotoxic effects of heavy metal contaminated soils. In the first part of the present study, the data base on metal effects in plant bioassays was extended. Four metal salts, namely Cr(VI)O3, Cr(III)Cl3, Ni(II)Cl2 and Sb(III)Cl3 were tested comparatively in MN tests with pollen tetrad cells of Tradescantia clone #4430 and in meristematic root tip cells of Vicia faba. With Cr6+ and Ni2+, clear-cut dose-effects were observed in a range between 0.75 and 10.0 mM, whereas this was not the case with Cr3+ (range tested 1.25-10 mM) and Sb3+ (range 0.30-5.25 mM). In Vicia, negative results were obtained with the four metal salts under all conditions of test. To compare the mutagenic potencies of the metals, the increases of the regression curves (k-values) were calculated, they indicate the number of MN induced per mM in 100 tetrad cells. The corresponding values for Cr6+ and Ni2+ are 0.87 and 1.05, respectively. It appears that the Tradescantia system is in particular sensitive towards those metal species which cause DNA damage in animals and man such as Cr6+, Cd2+, Ni2+, and Zn2+, whereas no clear positive results were obtained with less harmful metal ions such as Cu2+, Cr3+ or Sb3+. In the second part of the study, the mutagenic effects of four metal contaminated soils and two types of standardized leachates (pH 4.0 and pH 7.0) of these soils were tested in Tradescantia and in Vicia. In addition, chemical analyses were carried out to determine the metal concentrations in the soils and in the extracts. Two of the samples contained highly elevated levels of a number of metals (Zn, Pb, Cu, Cd, Sb, As), one soil came from the Central Austrian Alps and contained high As levels only. Direct exposure of the Tradescantia plants in the soils resulted in a drastic increase of the MN frequencies over the background. The lowest effect was seen with the Slovakian soil which contained in particular Sb and As (4.5-fold increase over the background), with the other soils, the induced frequencies were 11-15-fold over the control values. On the contrary, negative results were obtained upon exposure of Tradescantia cuttings in the leachates and upon implantation of germinated Vicia beans in the soils. The results of the present study indicate that Trad-MN assays with direct exposure of intact plants is an appropriate method which enables to detect genotoxic effects of metal contaminated soils in situ. This simple and fast biomonitoring assays might be a valuable supplement to analytical analyses of contaminated soils.

Antimony↗

Spectroscopic and ligand-field properties of [L-prolylglycinato][di(3-aminopropyl)amine]chromium(III) perchlorate.

The mixed ligand complex [Cr(L-progly)(dpt)]ClO4 [H2progly = prolylglycine, dpt = di(3-aminopropyl)amine] has been prepared, and its geometric structure was discussed on the basis of electronic absorption and IR spectral data. The emission and excitation spectra taken at 77 K were also reported. The 12 electronic bands due to spin-allowed and spin-forbidden transitions were assigned. Using the electronic transitions, a ligand-field optimization has been performed to determine the metal-ligand bonding properties for the coordinated atoms. We confirmed that the peptide nitrogen of prolylglycinate has moderate strong sigma- and weak pi-donor properties toward chromium(III) ion.

Chemical Phenomena↗

Intestinal absorption in health and disease: micronutrients.

The main theme of this chapter concerns the precise biochemical mechanisms involved in stages up to, and including, gastrointestinal absorption of vitamins and certain selected minerals. Essential data regarding sequential events, immediately following absorption of these micronutrients, are also included. There is reference to water-soluble vitamins that are, in general, either coenzymes in various metabolic reactions or carriers of certain biochemical groupings. In contrast, fat-soluble vitamins frequently function as integral components of cell membranes; they, too, receive ample attention. It is appropriate, nevertheless, to recognize that some minerals required in very small amounts are closely allied biochemically with particular vitamins; these specific associations are apportioned emphasis at relevant places in the text. Although predominant discussion centres on the physiological state, clinical reference is necessarily made to gastrointestinal disorders in which imbalance of vitamins and minerals consequently results in an additional detrimental impact on health.

Ascorbic Acid↗

The use of P-aminobenzoic acid and chromic oxide to confirm complete excreta collection in a carnivore, Felis silvestris catus.

Complete excreta collection is a pre-requisite for several protocols in protein metabolism, and lack of confidence in achieving this may be increased when working with carnivores. Recovery of p-aminobenzoic acid (PABA) as a check for complete urine collection and chromic oxide for complete faeces collection were assessed in the cat. A single oral dose of PABA (4 mg/kg BW) was excreted more slowly than has been reported in the human (82% recovery at 6 h). A daily dose of PABA proved a useful method for confirming complete urine collection in the cat, and was 99% excreted in 72 h. Chromic oxide (500 mg/cat) was administered orally and recovery of chromium in the faeces was 90% after 96 h. A HPLC method for the analysis of PABA in cat urine was developed, and from the application of the techniques to a nitrogen balance study, it was concluded that PABA and chromic oxide are useful checks for complete excreta collection in the cat.

4-Aminobenzoic Acid↗

The effect of dietary carbohydrates with different digestibility on the populations of Oesophagostomum dentatum in the intestinal tract of pigs.

An experiment was undertaken to study the effect of dietary carbohydrates with different digestibility on the populations of Oesophagostomum dentatum in the intestinal tract of pigs. Sixty-four worm-free pigs from a specific pathogen-free farm were randomly divided into 8 equal groups. The animals in 4 groups were assigned to a diet with partially undegradable carbohydrates (diet 1), while the pigs in the 4 remaining groups were given a diet with fermentable carbohydrates (diet 2). Diet 1 was comprised of barley flour, oat husk meal, soybean meal, vitamins and minerals and diet 2 of barley flour, inulin and sugar beet fibre, soybean meal, vitamins and minerals. The pigs in 6 of the groups (n = 48) were inoculated with 6,000 infective larvae of O. dentatum. To determine O. dentatum populations at the early stage of infection, 16 pigs were slaughtered 3 weeks p.i., while the remaining 4 groups continued on the diets for a further 9 weeks after which they were slaughtered. In a diet cross-over experiment 6 weeks after inoculation, 8 pigs changed from diet 1 to diet 2 (diet 1 > diet 2), and 8 pigs from diet 2 to diet 1 (diet 2 > diet 1). The results showed that partially undegradable carbohydrates provided favourable conditions not only for parasite establishment and sustainability, but also for already established O. dentatum infection while, in contrast, the diet composed of highly degradable carbohydrates decreased worm establishment, size and female fecundity. The implications for pastured pigs or pigs fed different complex carbohydrate diets is discussed.

Animals↗

Unsuitability of Cr(II) reduction for the measurement of sulfides in oxic water samples.

After developing a highly sensitive method for detecting acid-volatile sulfides (AVS) in oxic freshwaters, we hoped to apply that method to measuring a different class of dissolved reduced sulfur compounds, chromium-labile sulfides (CLS). A popular method for measuring this pool of sulfides in sediments relies on reduction dissolution of metal sulfides by Cr(II) and has been employed by researchers for over 15 years. Here, we demonstrate that this method is inappropriate for measuring CLS in oxic freshwaters in which sulfate concentrations are large relative to the dissolved metal sulfides. We observe the reduction of sulfate by Cr(II), and this presents a significant interference.

Chromium↗

Chromium(III) porphyrin as a selective ionophore in a salicylate-selective membrane electrode.

The construction, potentiometric response properties, and applications of a novel ion-selective electrode with high selectivity toward salicylate are described. Chromium(III) tetraphenylporphyrin chloride was used as ion carrier into plasticized PVC membrane. This ionophore is capable of serving as both a positively charged and neutral carrier, depending on the pH of the sample solution. The influence of several variables was investigated to optimize the potentiometric response and selectivity of the electrode. The resulting electrode demonstrates a near-Nernstian response over a wide range of salicylate concentration (10(-6)-10(-1) M). This electrode has a fast response time and micromolar detection limit and could be used over a wide pH range (3-9). The proposed electrode showed very high selectivity for salicylate over a number of common inorganic and organic anions. The specific interaction of salicylate with the central metal of porphyrin is described based on UV-visible absorption spectra. The electrode was successfully applied to the determination of salicylate in pharmaceutical preparations and clinical samples.

Aspirin↗

Reduction of chromium(VI) by ascorbate leads to chromium-DNA binding and DNA strand breaks in vitro.

Chromium(VI) is a known human carcinogen which requires intracellular reduction for activation. Ascorbate (vitamin C) has been reported to function as a major reductant of Cr(VI) in animals and cell culture systems. The reaction of Cr(VI) with varying concentrations of ascorbate was studied under physiological conditions in vitro in order to determine the types of reactive intermediates produced and to evaluate the reactivity of these intermediates with DNA. Reactions of 1.8 mM Cr(VI) with 0-18 mM ascorbate at pH 7.0 in N-(2-hydroxyethyl)piperazine-N'-2-ethanesulfonic acid (HEPES; 0.10 M) and tris(hydroxymethyl)aminomethane hydrochloride (Tris.HCl; 0.050 M) buffers were studied by electron paramagnetic resonance and UV/visible spectroscopy. Cr(V) and carbon-based free radical adducts of 5,5-dimethyl-1-pyrroline 1-oxide (DMPO) were observed at 0.5 to 1 and 1 to 1 reactions of ascorbate to Cr(VI). Levels of Cr(V) were higher for reactions in HEPES buffer, and levels of carbon-based radicals were higher in Tris.HCl buffer. Levels of Cr(IV) and Cr(III) increased with increasing concentration of ascorbate in both buffers. Reaction of Cr(VI) with varying ascorbate in the presence of calf thymus DNA or pBR322 DNA resulted in Cr-DNA adducts and plasmid relaxation, respectively. Maximum binding of Cr to DNA was observed for the 1:1 reaction ratio of Cr(VI) with ascorbate in both HEPES and Tris.HCl buffers, but total Cr bound to DNA was 8-fold lower in Tris.HCl than HEPES buffer. Preincubation of Cr(VI) with ascorbate before reaction with DNA decreased Cr-DNA binding to background levels. Preincubation of Cr(III) with ascorbate resulted in only low Cr-DNA binding. Levels of Cr-DNA binding were higher with single-stranded vs double-stranded DNA. Reactions with 14C-labeled ascorbate produced no cross-linking of ascorbate to DNA. Maximum plasmid relaxation was observed for the 1:1 ascorbate to Cr(VI) ratio in both buffers; however, single-strand breaks were 2-fold higher in Tris.HCl than HEPES buffer. Reactions with plasmid in the presence of DMPO quenched formation of single-strand breaks. Interpretation of these results in light of the spectroscopic studies suggested that Cr(V) and carbon-based radicals were responsible for Cr-DNA adducts and DNA single-strand breaks, respectively.

Ascorbic Acid↗

Chromium oligopeptide activates insulin receptor tyrosine kinase activity.

A possible new mechanism for the amplification of insulin receptor tyrosine kinase activity in response to insulin has been identified. The chromium-containing oligopeptide low molecular weight chromium-binding substance (LMWCr) does not effect the tyrosine protein kinase activity of rat adipocytic membrane fragments in the absence of insulin; however, insulin-stimulated kinase activity in the membrane fragments is increased up to 8-fold by the oligopeptide. Using isolated rat insulin receptor, LMWCr has been shown to bind to insulin-activated insulin receptor with a dissociation constant of circa 250 pM, resulting in the increase of its tyrosine protein kinase activity. The ability of LMWCr to stimulate insulin receptor tyrosine kinase activity is dependent on its chromium content. The results appear to explain the previously poorly understood relationship between chromium and adult-onset diabetes and cardiovascular disease.

Adipocytes↗