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Urinary chromium excretion of human subjects: effects of chromium supplementation and glucose loading.

The utilization of inorganic chromium by free-living human subjects was studied in 76 volunteers (male, 48; female, 28) who were supplemented with 200 micrograms of inorganic chromium as chromic chloride or a placebo tablet for 3 months in a double-blind, cross-over experiment. For all subjects, initial mean +/- SEM urinary chromium (Cr) level was 0.20 +/- 0.01 (range, 0.05 to 0.58) ng/ml and did not differ by sex. Initial chromium/creatinine ratio (Cr/Ct) was 0.15 +/- 0.01 (range 0.03 to 0.36) ng Cr/mg creatinine for females and was significantly lower, 0.10 +/- 0.01 (range 0.03 to 0.36) for males. Mean urinary Cr level increased to 1.0 +/- 0.12 after 2 and to 1.13 +/- 0.08 ng/ml after 3 months' supplementation. The Cr/Ct ratio increased to 0.69 +/- 0.10 for females and to 0.50 +/- 0.04 for males after 2 months' supplementation; values were similar after 3 months. An increase in urinary Cr excretion in response to a glucose load was demonstrated for nonsupplemented normal free-living subjects but not for subjects supplemented daily with trivalent chromium. Urinary Cr excretion after a glucose challenge was not predictable and did not depend on Cr status.

Adult↗

Effect of inorganic chromium supplementation on glucose tolerance, insulin response, and serum lipids in noninsulin-dependent diabetics.

A placebo-controlled double-blind cross-over study was carried out to assess the effect of chromium supplementation (200 micrograms trivalent chromium daily for 6 wk) on glucose tolerance, insulin response, long-term diabetic control, and serum lipids in 10 noninsulin-dependent diabetics aged 37 to 68 yr. After chromium supplementation 24-h urinary chromium excretion showed a 9-fold increase indicating a positive chromium balance in the subjects. There was no significant difference between chromium supplementation and placebo periods in glucose tolerance and in fasting or 2-h postglucose serum insulin levels but the 1-h postglucose serum insulin level was slightly lower on chromium supplementation than on the placebo (55 +/- 9.0 versus 64 +/- 11; p less than 0.01, paired t test). Serum total cholesterol and triglycerides and their high-density, low-density, and very low-density lipoprotein subfractions showed no change after chromium supplementation as compared to the placebo period.

Adult↗

The effects of inorganic chromium and brewer's yeast on glucose tolerance, plasma lipids, and plasma chromium in elderly subjects.

Twenty-three healthy, well-nourished, free-living elderly volunteers were given daily, for 10 wk, 5 g brewer's yeast, 200 micrograms Cr3+ as chromic chloride (CrCl3), or placebo. There were no significant changes in glucose tolerance, insulin, cholesterol or triglycerides after supplementation in any of the three groups. Plasma Cr3+ rose significantly after supplementation with CrCl3 but did not change after yeast or placebo supplementation. Plasma Cr3+ did not increase after an oral glucose load and did not correlate with glucose, insulin or lipid values in any of the groups. Calculated intakes of eight indicator nutrients were well above 100% of the RDA except for calcium. These healthy elderly persons, eating nutritious diets, are not at risk for Cr3+ deficiency as measured by the absence of a clinical response to CrCl3 or brewer's yeast supplementation. This study suggests that age per se is not a factor leading to Cr deficiency.

Aged↗

An improved method for oxidation of chromium(III) oxide-containing fecal samples by using sodium peroxide fusion.

A safer method of oxidation of Cr2O3-containing fecal samples from transit-time studies was developed using sodium peroxide to replace perchloric acid as the oxidizing agent. The percentage recovery of Cr2O3 with this method was compared with that of perchloric acid method for samples containing quantities of fecal ash and Cr2O3 typical of those from rodent transit-time studies. Both methods gave relatively constant percentage recoveries for Cr2O3 contents from 0.4 to 10 mg. Over this range, mean (+/- SD) percentage recoveries of Cr2O3 for sodium peroxide fusion and the perchloric acid method were 75.5 +/- 4.3 and 89.9 +/- 2.5, respectively. As long as percentage recovery is constant, the transit time as determined by calculation of the time of 80% excretion of the total recovered Cr2O3 is not affected. Sodium peroxide fusion provides a useful and safer alternative to perchloric acid oxidation in transit-time studies using Cr2O3 as a nonabsorbable marker.

Animals↗

Transformation of non-tumorigenic osteoblast-like human osteosarcoma cells by hexavalent chromates: alteration of morphology, induction of anchorage-independence and proteolytic function.

Soluble chromium (VI) compounds either alone or in combination with 3-methylcholanthrene (MC) were used to transform non-tumorigenic osteoblast-like human osteosarcoma cells (HOS TE85). The Cr(VI) compounds were highly toxic to these cells with LC50 values in the range of approximately 0.5-1.0 microM. Continuous passaging of the treated cells resulted in sustained increase in anchorage-independent (AI) colony formation. Treatment with Cr(VI) and MC resulted in substantial increase in AI growth. At the XVth passage, a number of individual AI colonies were expanded in culture and used for further studies. The cells are refractory in appearance and grow as 'nests' rather than as monolayers. The cell lines have relatively high plating efficiency (PE) in soft agar and respond to promotional effect of phorbol-12-myristate-13-acetate by an increase in PE and in the size and number of AI colonies. While the isolated cells are not tumorigenic when tested in athymic nude mice, most of the lines possess higher levels of plasminogen activator (PA) activity, considered as one of the markers of transformation. This is also reflected in the increase in the steady state level of urokinase type PA mRNA. These results show that Cr(VI) compounds are capable of promoting human cells to an altered phenotype characteristic of a stage in the carcinogenesis cascade.

Animals↗

Differential binding of chromium(VI) and chromium(III) complexes to salmon sperm nuclei and nuclear DNA and isolated calf thymus DNA.

The binding of CrCl3.6H2O, Cr(NO3)3.9H2O, [Cr(L-His)2] (NO3)3.H2O, [Cr(L-Cys)(L-His)].3.5H2O, [Cr(L-His)(D-Pen)].H2O, Na[Cr(L-Cys)2].2H2O, K2[Cr(GS)2].3H2O, Na2-CrO4.4H2O, and Na2Cr2O7.2H2O to salmon sperm nuclei and nuclear DNA was determined. The Cr(III)-amino acid complexes and Cr(VI) exhibited significantly lower Cr-nuclei and Cr-DNA binding levels relative to the inorganic complexes CrCl3.6H2O and Cr(NO3)3.9H2O. The binding of CrCl3.6H2O, Cr(NO3)3.9H2O and Na2Cr2O7.2H2O to salmon sperm nuclei and nuclear DNA in the presence of rat lung cytosol was determined under the same conditions. For those complexes studied in both buffer and cytosol, the Cr-DNA binding levels for Cr(III) complexes were higher in buffer than in cytosol, while a relatively higher binding level was observed for Cr(VI) in cytosol than in buffer. Slightly lower nuclear protein levels were present in Cr(VI) incubations than in Cr(III) incubations with nuclei both in the presence and the absence of cytosol. The relative binding of CrCl3.6H2O, Cr(NO3)3.9H2O, [Cr(L-His)2](NO)3.H2O, [Cr(L-Cys) (L-His)].3.5H2O, [Cr(L-His)(D-Pen)].H2O, Na[Cr(L-Cys)2].2H2O and Na2CrO4.4H2O to isolated calf thymus DNA in buffer was also determined. Positively charged, labile inorganic Cr(III) complexes, CrCl3.6H2O and Cr(NO3)3.9H2O, exhibited higher binding to DNA than [Cr(L-His) (D-Pen)].H2O, and no binding to DNA was observed with Cr(VI) and the other neutral, positively and negatively charged, inert Cr(III)-amino acid complexes. Although labile aquo chromium(III) complexes are quite reactive with DNA, the reactivity of chromium(III), formed upon intracellular reduction of carcinogenic chromium(VI), toward DNA will be diminished by complexation with cellular proteins, peptides and amino acids.

Amino Acids↗

Two pathways for chromium(VI)-induced DNA damage in 14 day chick embryos: Cr-DNA binding in liver and 8-oxo-2'-deoxyguanosine in red blood cells.

Previous studies have shown that chromium(VI) induced cell-specific types of DNA damage, i.e. DNA cross-links in liver and DNA strand breaks in red blood cells, in 14 day chick embryos. Direct and indirect pathways for chromium(VI)-mediated DNA damage, in the form of Cr-DNA binding and 8-oxo-2'-deoxyguanosine (8-oxo-dG) respectively, were examined in liver and red blood cells of 14 day chick embryos. Levels of hepatic Cr-DNA binding increased in a Cr(VI) dose-dependent manner. Cr-DNA binding in red blood cells was 10-fold lower than in liver, although the Cr-uptake in red blood cells was only 2-fold lower than in liver. The level of 8-oxo-dG formation in red blood cells increased at all Cr(VI) doses tested but peaked at 0.10 mmol Cr(VI)/kg, whereas no increase in 8-oxo-dG levels over background levels was observed in liver of Cr(VI)-treated embryos. The possible role of glutathione in modulating Cr(VI)-induced DNA damage was examined by using L-buthionine-R,S-sulfoximine (BSO) to deplete glutathione. No changes in glutathione levels were observed in either liver or red blood cells of embryos treated with Cr(VI) in the presence or absence of BSO pretreatment. Ascorbate levels in liver and red blood cells were not affected by treatment of embryos with chromium(VI), BSO or Cr(VI) and BSO. Depletion of glutathione by BSO resulted in a small increase of chromium uptake in liver of embryos treated with 0.050 and 0.10 mmol Cr(VI)/kg, but had no effect on hepatic chromium uptake at 0.20 mmol Cr(VI)/kg. BSO had no effect on chromium uptake in red blood cells. Depletion of glutathione had no effect on hepatic or red blood cells Cr-DNA binding in embryos treated with Cr(VI). However, depletion of glutathione significantly decreased the 8-oxo-dG levels in red blood cells at all Cr(VI) doses tested. Levels of 8-oxo-dG in liver of Cr(VI)-treated embryos remained at background in the presence or absence of BSO pretreatment. These results indicate that Cr(VI)-induced DNA damage in 14 day chick embryos is through a direct interaction of chromium with DNA in liver, but is through an indirect oxidative pathway in red blood cells. It appears that glutathione plays an important role in chromium(VI)-induced formation of 8-oxo-dG in red blood cells.

8-Hydroxy-2'-Deoxyguanosine↗

High mobility group proteins 1 and 2 recognize chromium-damaged DNA.

Chromium (Cr) is a human carcinogen and a potent DNA damaging agent. Incubation of DNA with CrCl3 resulted in dose-dependent binding of Cr to DNA and, at concentrations >20 microM, altered the electrophoretic mobility of a 100 bp oligonucleotide. We also demonstrate that high mobility group (HMG) proteins 1 and 2 bind Cr-damaged DNA (Cr-DNA). Protein binding was lesion density-dependent, with maximal binding to DNA treated with 100 microM CrCl3. HMG2 binds to Cr-DNA with a calculated Kd of approximately 10(-9) M. These proteins also bound DNA obtained from chromate-treated cells. These results suggest that the covalent attachment of Cr to DNA induces alterations in DNA structure which are recognized by HMG1 and HMG2. Therefore, these proteins may function as Cr-damaged DNA recognition proteins in vivo and as a consequence of binding, may play a role in directing the cellular response to Cr-DNA adduct formation.

Animals↗

A comparison of a microdiffusion method for ethanol and the Abbott TDx ethanol assay.

A simple, rapid ethanol screen based on a microdiffusion technique (Analytical Systems, Marion Laboratories) has been evaluated and compared to an established enzymatic procedure (Abbott TDx). They showed agreement in 484 of 488 biological specimens and all control specimens. They disagreed on 4 specimens--3 positive by the Marion method and negative by TDx, and 1 positive by TDx and negative by Marion. The discrepancies could not be elucidated because insufficient quantities of samples remained. The microdiffusion method was found to be a dependable, reproducible screening test for the presence of reducing volatiles in urine. There is insufficient data to evaluate its effectiveness in serum or whole blood. It is particularly suited for screening negative specimens before quantitating positive specimens by a suitable alternate quantitative method.

Chromium↗

Carbohydrate utilization by tilapia (Oreochromis niloticus x O. aureus) as influenced by different chromium sources.

An experiment was conducted to study the influence of three different forms of chromium (CrCl3.6H2O, Na2CrO4.4H2O and Cr2O3) on the utilization of two carbohydrates (glucose and cornstarch) by juvenile hybrid tilapia (Oreochromis niloticus x O. aureus). Average initial body weight of the fish was 1.13 +/- 0.02 g. Fish were fed 5% body wt/d. Significantly (P < 0.05) greater body weight gain, food intake, protein retention, energy retention and body lipid concentration were observed in fish fed the starch diet than in those fed the glucose diet. Fish fed the glucose diet supplemented with any type of chromium had significantly greater weight gain than those fed the glucose diet without chromium supplementation. Fish fed the glucose diet supplemented with Cr2O3 had greater weight gain, food intake, protein retention, energy retention and body lipid concentration than those fed the unsupplemented glucose diet or the glucose diet supplemented with CrCl3.6H2O or Na2CrO4.4H2O. Delayed plasma glucose peak time was observed in tilapia fed the glucose diet supplemented with any type of chromium. Chromium supplementation generally lowered the glucose-6-phosphatase activity in tilapia. Phosphofructokinase activity was significantly higher in fish fed the glucose diet supplemented with Cr2O3 than in the other glucose-fed groups. These data suggest that chromium supplementation improved glucose utilization by tilapia and that Cr2O3 supplementation was markedly more effective than other chromium forms.

Animals↗

Carbohydrate utilization and digestibility by tilapia, Oreochromis niloticus x O. aureus, are affected by chromic oxide inclusion in the diet.

A 12-wk feeding trial was conducted to study the influence of chromic oxide (Cr2O3) on carbohydrate utilization and digestibility by tilapia, Oreochromis niloticus x O. aureus. Two levels of chromic oxide (0.5 and 2%) were each incorporated into diets containing glucose or starch. Chromic oxide was added at 0 or 8 wk. The diets were fed to triplicate groups of fish weighing 1.11 +/- 0.05 g. Fish fed the starch diet had greater (P < 0.05) weight gain, feed efficiency ratio, protein efficiency ratio, protein deposition and digestibility of protein, lipid, carbohydrate and dry matter than fish fed the glucose diet irrespective of the time and level of chromic oxide supplementation. Fish fed the glucose diet with 0.5% chromic oxide had higher weight gain, feed efficiency ratio, protein efficiency ratio and protein deposition than fish fed the glucose diet with 2% chromic oxide. The ingredient digestibility estimated using 0.5% chromic oxide as the marker was greater than that estimated with 2% chromic oxide. Higher phosphofructokinase and lower glucose-6-phosphatase activity was found in fish fed the starch diet than in fish fed the glucose diet regardless of the time and level of chromic oxide inclusion. Fish fed the glucose diet with 0.5% chromic oxide had higher phosphofructokinase activity and lower tissue chromium concentration than fish fed the glucose diet with 2% chromic oxide irrespective of chromic oxide inclusion time. These data suggest that the level of chromic oxide in the diet alters glucose utilization by tilapia and affects nutrient digestibility by tilapia. The time of chromic oxide inclusion had no effect on carbohydrate utilization and digestibility.

Animal Feed↗

Chromic oxide inclusion in the diet does not affect glucose utilization or chromium retention by channel catfish, Ictalurus punctatus.

This study was conducted to determine if the level of dietary chromic oxide will affect glucose utilization and tissue chromium retention by channel catfish. Purified diets containing graded levels of supplemental chromic oxide (0, 50, 100, 200, 400, 1000, 5000 and 10,000 mg/kg diet) and glucose as the carbohydrate source were fed to channel catfish fingerlings for 10 wk. Another diet containing dextrin as the carbohydrate source and without chromic oxide supplementation was also fed and served as the control diet. Fish fed the dextrin diet had significantly (P < 0.05) greater weight gain, feed efficiency ratio and protein efficiency ratio but lower plasma glucose concentrations than fish fed the glucose diets irrespective of the level of chromic oxide supplementation. The growth performance and postprandial plasma glucose concentrations of channel catfish fed glucose diets supplemented with various chromic oxide levels were not significantly different. No obvious trends were observed in the whole-body composition of fish fed glucose diets containing various chromic oxide levels. Carbohydrate source or the level of dietary chromic oxide did not significantly affect chromium concentrations in the whole-fish carcass. These results suggest that the level of dietary chromic oxide had no significant effect on glucose utilization or chromium retention by channel catfish. It is suggested that chromic oxide is sufficiently inert to be used as an external marker in digestibility studies in channel catfish.

Animals↗

Recovery of 15N-lactoferrin is higher than that of 15N-casein in the small intestine of suckling, but not adult miniature pigs.

Performance of biological functions of lactoferrin in the small intestine requires at least some resistance to degradation. Therefore, we studied prececal digestibility of lactoferrin in comparison to casein both in suckling and adult miniature pigs, applying 15N-labeled proteins. In study 1, 43 piglets (10-d-old), deprived of food for 12 h received 10 mL of sow's milk supplemented with 120 mg of 15N-labeled protein (porcine or bovine lactoferrin or bovine casein). Piglets were anesthetized 150 min later, after which the small intestine was excised, cut into three sections, and chyme was collected. In study 2, nine food-deprived boars fitted with T-canulae at the terminal ileum were given two semisynthetic experimental meals (204 g) in a cross-over design, 2 wk apart. One contained 7.5% (g/100 g) 15N-labeled bovine casein, the other 1.25% 15N-labeled bovine lactoferrin. Both were adjusted to 15% total protein with nonlabeled casein. Ileal chyme was collected from the canula over 33 h postprandially. All diets contained the indigestible marker chromic oxide. 15N-digestibility of lactoferrin, both porcine (84.4 +/- 3.2%) and bovine (82.3 +/- 4.8%), was significantly lower than casein digestibility (97.6 +/- 0.5%) in the distal small intestine of suckling piglets (P < 0.05). Based on immunoblotting after acrylamide electrophoresis, 4.5% of non- and partially digested lactoferrin was found in the last third of the small intestine of piglets. In adult miniature pigs there was no difference in 15N-digestibility of bovine lactoferrin compared to bovine casein (90.7 +/- 1.9% vs. 93.9 +/- 1.0%, P > 0.05). In suckling miniature pigs, the reduced digestibility of lactoferrin may provide the prerequisite for biological actions along the whole intestinal tract. The source of lactoferrin, porcine or bovine, made no difference in this respect.

Aging↗