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Enhancement of nucleolar RNA synthesis by chromium (III) in regenerating rat liver.

Rats pretreated with CrCl3 at an intraperitoneal dose of 5 mg Cr/kg body weight, and then subjected to partial hepatectomy concentrated Cr in the regenerating liver cells, especially in the nucleoli. Hepatic RNA synthesis induced by partial hepatectomy was further enhanced by Cr(III) administration without changes of the nucleotide pool size at least for 12 hr after the hepatectomy. Of the RNA species newly synthesized, the synthesis of nucleolar RNA was predominantly enhanced. The nucleolar RNA thus synthesized in the regenerating liver of Cr(III)-dosed rats followed normal processing into rRNA as estimated by actinomycin D-chase experiments followed by sucrose density gradient centrifugation analysis and also by nucleotide composition analysis of nucleolar and ribosomal RNAs. These results suggest that Cr accumulated in nucleoli may participate in nucleolar gene expression.

Animals↗

Insulin-like activity of chromium-binding fractions from brewer's yeast.

51CrCl3 was added to the incubation medium of Saccharomyces cerevisiae for up to 48 hr. After repeated freezing and thawing, lysing in 9 M urea with 1% NP-40 detergent, and dialysis against water, the lower molecular weight (Mr less than 3500) dialysate was retained on a SE53 cationic exchange column, eluted with 0.25 M NH4OH and fractionated on a Bio-gel P-2 column. The insulin-like biological activity of the fractions was measured by the 14C-glucose oxidation in isolated rat adipocytes. The biological activity that was found in two of nine fractions did not correspond to their chromium content. Moreover, identical findings were obtained when chromium was added not to the live yeast but to the yeast extract, which showed that its binding was a chemical process not requiring cellular activity. No fraction demonstrated insulin-potentiating activity on rat adipocytes.

Adipose Tissue↗

Protective effect of chromium(III) on acute lethal toxicity of carbon tetrachloride in rats and mice.

Trivalent chromium (Cr(III)) preadministered intraperitoneally (5 mg Cr/kg body weight) to rats and mice protected these animals from acute lethal toxicity of carbon tetrachloride (CCl4). Some other metals, Cr(VI), Cu(II), and Zn(II), had no effect on CCl4 lethal toxicity. DL-alpha-tocopherol, one of the antioxidative agents, showed similar protective effects to Cr(III). Activities of serum GOT and GPT in mice were increased sharply by the administration of CCl4, but these elevations were depressed by Cr(III) preadministration. Serum glucose levels of mice increased transiently after CCl4 administration and then in the control group fell to hypoglycemic levels after 6 hr, whereas the Cr(III)-pretreated group kept to homeostatic levels. Lipid peroxidation of microsomes in mice 24 hr after Cr(III) administration was lower than that of the control. These results suggest that Cr(III) preadministered to mice might act as a radical scavenger to CCl4 to form trichloromethyl radicals which are a major initial product of CCl4 in liver cells.

Animals↗

Chromium picolinate increases membrane fluidity and rate of insulin internalization.

The effects of chromium chloride, chromium nicotinate, and chromium picolinate on insulin internalization in cultured rat skeletal muscle cells was examined. Insulin internalization was markedly increased in cells cultured in a medium that contained chromium picolinate and the increased internalization rate was accompanied by a marked increase in the uptake of both glucose and leucine. The effect was specific for chromium picolinate since neither zinc picolinate nor any of the other forms of chromium tested was effective. The increased insulin internalization rate may result from an increase in membrane fluidity since chromium picolinate and to a lesser extent, chromium nicotinate, increased the membrane fluidity of synthetic liposomal membranes.

Animals↗

Clastogenicity of lead chromate particles in hamster and human cells.

Several insoluble compounds of chromium, such as lead chromate, are respiratory carcinogens in experimental animals and suspected to be so in humans. Lead chromate induces neoplastic transformation in cultured cells but the mechanism of genotoxicity is unknown. We examined the effect of lead chromate on the integrity of chromosomes of Chinese hamster ovary (CHO) and human foreskin fibroblasts (HFF) after a 24-h exposure. At 0.4 microgram/cm2, 0.8 microgram/cm2, 2 microgram/cm2 and 8 microgram/cm2 lead chromate particles reduced survival of CHO cells to 86%, 62%, 2% and less than 1% respectively. These concentrations induced a dose-dependent 4-19-fold increase in the percent metaphases with damage. The HFF cells exhibited higher sensitivity in both cytotoxicity and clastogenicity. The spectrum of damage observed for both cell types was primarily achromatic lesions affecting one or both chromatids. To test for particle dissolution effects, CHO cells were treated for 24 h with either clarified medium that had been incubated for 24 h with lead chromate particles, or clarified medium that had been pre-conditioned by CHO cells treated with lead chromate particles for 24 h. No damage was detected in these cells, indicating that extracellular dissolution into ionic lead and chromate did not contribute to the genotoxicity. This is consistent with a previous study in which scanning electron micrographs illustrated internalization of the particles. These results suggest that clastogenesis may be a mechanism for lead chromate induced carcinogenesis.

Animals↗

Chromium chloride induces chromosomal aberrations in human lymphocytes via indirect action.

The aim of this study was to examine the possible clastogenic effects of trivalent chromium chloride (CrCl3) as the results in the literature are non-conclusive. Under the conditions used in this study Cr(III) induces chromosomal aberrations in phytohemagglutinin(PHA)-stimulated human lymphocytes. This activity, however, is suppressed by the antioxidants superoxide dismutase (SOD) (scavenger of O-.2), the SOD-like agents, catalase and mannitol (specific scavenger of OH.). The possibility that oxygen free radicals could evolve through stimulation of the arachidonic acid cascade is suggested using suitable inhibitors.

5,8,11,14-Eicosatetraynoic Acid↗

Differential protection of chlorophyllin against clastogenic effects of chromium and chlordane in mouse bone marrow in vivo.

The anticlastogenic activity of sodium-copper-chlorophyllin was tested against two known clastogens, chromium(VI) oxide and the pesticide chlordane, both of which occur as environmental toxicants. The chemicals were administered to male Swiss albino mice by gavage and chromosomes were studied from bone marrow cells 24 h after the exposure, following colchicine-air drying-Giemsa preparation. The end-points screened were the frequencies of chromosomal aberrations and damaged cells. The clastogenic effects induced by chromium in aqueous solution (20 mg/kg body weight) were reduced to a significant level by chlorophyllin (1.5 mg/kg body weight), when administered both before and simultaneously with the toxicant. The effects of chlordane in olive oil (10 mg/kg body weight) were, however, not reduced by the administration of chlorophyllin (1.5 and 3 mg/kg body weight).

Analysis of Variance↗

Evaluation of methods for chemically coupling foot-and-mouth disease virus to sheep red blood cells for immunological assays.

Six methods of chemically coupling proteins to red blood cells were evaluated for their effectiveness in coupling foot-and-mouth disease virus (FMDV) to sheep red blood cells. The coupling agents tested were potassium periodate, 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (ECDI), chromium chloride, glutaraldehyde, bis-diazotized benzidine (BDB) and N-succinimidyl 3-(2-pyridyldithio) propionate (SPDP). Of these, only the coupling methods using BDB and SPDP resulted in virus-red cell complexes that reacted with FMDV antiserum in passive hemagglutination and passive immune hemolysis assays. The BDB and SPDP methods were studied further to determine optimal coupling conditions, the kinetics of coupling and the effects of chemical couplers on viral integrity. Only the FMDV-red cell complexes formed with SPDP were suitable targets for detecting FMDV antibody producing lymphocytes in a hemolytic plaque assay.

Animals↗

Effects of inhaled chromium dioxide dust on rats exposed for two years.

Rats were exposed by inhalation to chromium dioxide (CrO2) dust at design concentrations of 0, 0.5 mg/m3 (stabilized and unstabilized, respectively), or 25 mg/m3 (stabilized) for 6 hr/day, 5 days/week for 2 years. No dust-exposure-related pathological changes other than lung lesions were observed in all exposed rats. There were no significant differences in pulmonary responses between unstabilized and stabilized CrO2 at the 0.5 mg/m3 exposure level. The lungs showed minute dust deposition in the alveoli adjacent to the alveolar ducts, but maintained an intact general architecture. The pulmonary responses satisfied the biological criteria for a nuisance dust. At 25 mg/m3, dust deposition was sharply confined to the alveoli in the alveolar duct region. Alveolar walls enclosing dust-laden macrophage (dust cell) aggregates were thickened with hyperplastic Type II pneumocytes and slightly collagenized fibrosis. Alveoli adjacent to the terminal bronchioles were lined with bronchiolar epithelium (alveolar bronchiolarization). In addition, lungs showed foamy macrophage response, cholesterol granulomas, alveolar proteinosis, and minute fibrotic pleurisy. These pulmonary lesions occurred predominantly in female rats. Of 108 female rats, 6 developed keratin cysts and 2 had cystic keratinizing squamous cell carcinoma (CKSCC). None of 106 male rats had either a keratin cyst or a CKSCC. The lung tumors developed from metaplastic squamous cells in the areas of alveolar bronchiolarization at the alveolar duct region. The lung tumors were well differentiated and devoid of characteristics of true malignancy. The CKSCC is an experimentally induced, unique tumor type and is different from the type of spontaneous lung tumor seen in man or animals. The relevance to man of this type of lung tumor appears to be negligible.

Aerosols↗

Transfer of chromic chloride to embryonic mice and changes in the embryonic mouse neuroepithelium.

Transfer of CrCl3 to embryonic mice and changes in the embryonic mouse neuroepithelium associated with Cr-induced neural tube defects were studied. A single intraperitoneal injection of 51CrCl3 was administered on the 8th day of gestation and animals were killed at various intervals for measurements of radioactivity in the maternal blood and embryos. Radioactivity per gram of fetal tissues increased during the observed period, while those of maternal blood decreased. Pregnant mice were injected intraperitoneally with a single dose of CrCl3 (non-radioactive) on the 8th day of gestation. Animals were killed at specified times after injection and embryos were removed and examined histologically. At 4, 8 and 12 h after injection many pyknotic cells were observed on the neural plate, especially at 8 h after injection. These data suggest that CrCl3 may affect embryos directly and cause neural tube defects and that pyknotic cells on the neural plate may be an early pathological finding in cases of developing exencephaly.

Animals↗

Carcinogenicity of sodium dichromate and chromium (VI/III) oxide aerosols inhaled by male Wistar rats.

In inhalation chambers, male Wistar rats of the strain TNO-W74 were continuously exposed to submicron aerosols of sodium dichromate and to a pyrolyzed Cr(VI)/Cr(III) (3:2) oxide mixture. The sodium dichromate (Na2Cr2O7) aerosol had the chromium concentrations of 25, 50 and 100 micrograms/m3, the chromium oxide mixture (Cr5O12) had the chromium concentration of 100 micrograms/m3. After 18 months of inhalation the rats were held under conventional conditions for a further year. The experimental groups consisted of 20 rats and the control group of 40 rats. More than 90% of the rats in each group reached 2 years. At the end of the study the mortality rates amounted to 35%, 45% and 25% in the 3 sodium dichromate aerosol groups, respectively, and 50% in the chromium oxide mixture aerosol group, which was not significantly different from that of the controls (42.5%), living under the same conditions in filtered fresh air. In all sodium dichromate exposed groups significant effects were neither found clinically nor from hematology and clinical chemistry compared to the controls. In the chromium oxide mixture group, however, there was a number of significant findings. Elevated white and red blood cell counts and serum cholesterol as well as decreased serum total immunoglobulin levels at different stages of the study were observed together with few local lung effects determined histopathologically in this group. We assume that these effects are mainly due to the increased chromium lung burden of the rats. At the end of the study the lung chromium retention was about 10 times higher for the rats exposed to chromium oxide versus sodium dichromate at an aerosol Cr-concentration of 100 micrograms/m3, while the kidney chromium retention was measured to be nearly equal in both groups. Three primary lung tumors (2 adenomas and 1 adenocarcinoma) and 1 malign tumor of the pharynx were found at the highest Cr-concentration (100 micrograms/m3) of the sodium dichromate aerosol, 1 primary adenoma of the lung was in the chromium oxide mixture group exposed also to a Cr-concentration of 100 micrograms/m3. No primary lung tumors were observed in the other experimental and control groups. These results indicate a weak carcinogenicity at 100 micrograms/m3 for the rats continuously exposed to submicron Na2Cr2O7 and Cr5O12 aerosols. Thus, there may be a small carcinogenic risk from occupational relevant chromium air levels. However, results have to be confirmed with larger animal populations.

Administration, Inhalation↗

The use of inert markers in the measurement of the digestibility of cubed concentrates and of hay given in several proportions to the pony, horse and white rhinoceros (Diceros simus).

1. The rate of passage of chromic oxide (Cr) was similar in the pony and rhinoceros. 2. Higher apparent amounts digested were found using 4N-HCl-insoluble ash (AIA) than those determined by Cr, but overall digestibilities were similar for the two species. 3. An abrupt increase in the starch content of the horse diets increased the number of faecal ciliate protozoa. Only when the overall feed intake was increased in horses receiving a high dietary proportion of starch were the numbers depressed. 4. When the rhinoceros received 109 kJ apparent DE per kg bodyweight daily (716 kJ/W0.75 daily) it maintained normal condition.

Animal Feed↗

Chromic gut suture reduces calcitonin-gene-related peptide and substance P levels in the spinal cord following chronic constriction injury in the rat.

The chronic constriction injury (CCI) is an animal model of an experimental peripheral neuropathy. In this model, a mononeuropathy is produced by loosely ligating the left sciatic nerve of the rat with chromic gut suture (Bennett and Xie 1988). Maves et al. (1993) have proposed that chemical constituents of chromic gut suture influence the behavioral changes of rats with the CCI. Considering their results, we became interested in evaluating whether the type of suture material used to produce the CCI also affected spinal levels of calcitonin-gene-related peptide immunoreactivity (CGRP-ir) and substance P immunoreactivity (SP-ir), peptides that are associated with small primary afferent neurons. Using methods of radioimmunoassay (RIA), we measured levels of CGRP-ir and SP-ir in the dorsal quadrants of approximately the lumbar 4-5 (L4-L5) spinal segments of rats with a CCI induced using polyglactin (Vicryl), plain gut, or chromic gut suture. We observed bilateral decreases in CGRP-ir and SP-ir 60 days after a CCI induced with chromic gut suture, but no changes in peptide levels after a CCI induced with either polyglactin or plain gut suture. These results suggest two possibilities: (1) chromic gut suture, when used to produce the CCI, has more than just a constrictive effect on the sciatic nerve, and/or (2) different suture materials produce changes in CGRP-ir and SP-ir with a differential time-course. Our experiments are unable to distinguish between these two possibilities.

Animals↗

Isolation and structural studies of the neutral oligosaccharide units from bovine glycophorin.

The O-glycosidically linked carbohydrate units of glycophorin from bovine erythrocyte membranes were released as reduced oligosaccharides by alkaline borohydride treatment. These oligosaccharides were separated by ion-exchange chromatography followed by gel filtration. Three oligosaccharides, a penta- a hepta- and a decasaccharide were obtained as the major components from the neutral fraction, and seven fractions were separated from the acidic fractions. All of the fractions were found to contain galactose and N-acetylglucosamine in variable amounts, as well as N-acetylgalactosaminitol. Studies of the neutral oligosaccharides by methylation analyses, nitrous acid deamination and Smith degradation, indicated the structure of the pentasaccharide to be Gal(1 leads to 3)Gal(1 leads to 4)GlcNAc(1 leads to 3)GalNAcol and that of the heptasaccharide to be Gal(1 leads to 3)Gal(1 leads to 4)GlcNAc(1 leads to 3)Gal(1 leads to 4)GlcNAc(1 leads to 3)Gal(1 leads to 3)GalNAcol. The highest molecular weight fraction, decasaccharide in the neutral fraction had a branching point at C-6 of a galactose residue.

Animals↗