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Reduction and precipitation of chromate by mixed culture sulphate-reducing bacterial biofilms.

The ability of sulphate-reducing bacterial biofilms to reduce hexavalent chromium (Cr(VI)) to insoluble Cr(III), a process of environmental and biotechnological significance, was investigated. The reduction of chromate to insoluble form has been quantified and the effects of chromate on the carbon source utilization and sulphate-reducing activity of the bacterial biofilms evaluated. Using lactate as the carbon/energy source and in the presence of sulphate, reduction of 500 micromol l-1 Cr(VI) was monitored over a 48-h period where 88% of the total chromium was removed from solution. Mass balance calculations showed that ca 80% of the total chromium was precipitated out of solution with the bacterial biofilm retaining less than 10% of the chromium. Only ca 12% of the chromate added was not reduced to insoluble form. Although Cr(VI) did not have a significant effect on C source utilization, sulphate reduction was severely inhibited by 500 micromol-1 Cr(VI) and only ca 10% of the sulphate reducing activity detected in control biofilms occurred in the presence of Cr(VI). Low levels of sulphide were also produced in the presence of chromate, with control biofilms producing over 10-times more sulphide than Cr(VI)-exposed biofilms. Sulphide- or other chemically-mediated Cr(VI) reduction was not detected. The biological mechanism of Cr(VI) reduction is likely to be similar to that found in other sulphate-reducing bacteria.

Biodegradation, Environmental↗

Cancer mortality in a pigment plant utilizing lead and zinc chromates.

Several studies of workers exposed to various forms of chromium compounds have suggested an increased incidence of respiratory cancers. Lead and zinc chromates were among the chromium compounds implicated. The Department of Preventive Medicine and Community Health of the New Jersey Medical School undertook a detailed mortality study of a pigment plant in Newark which utilized both of these compounds. We compared observed deaths from each cause among 1296 white and 650 non-white males who were employed at the plant between January 1, 1940 and December 31, 1969, with expected deaths, as computed from cause-, age-, and time-specific standard death rates for the United States. A statistically significant relative risk of 1.6 for lung cancer among white male employees was found, as well as among the cohorts of white males employed 10 yr or more. A relative risk of 1.9 was noted for individuals employed at least 2 yr who were at least moderately exposed to chromates. An increased incidence of lung cancer among non-white males and stomach and pancreatic cancer among the total cohort was also evident. These data support the validity of the association between chromate pigment exposure and lung cancer, suggesting that pigment workers and other exposed to chromates be observed carefully in regard to possible risk or pancreatic and stomach cancers.

Black People↗

Selective removal of chromates by macroporous exchanger Amberlyst A-21.

Sorption studies of chromate on weakly basic macroporous anion exchanger Amberlyst A-21 were performed as a function of pH (2 - 9), temperature (20 - 40 degrees C) and concentration (0.4 - 80 mmol.l(-1)). Chromate sorption was observed to decrease with the increase in initial pH of the solution, with a sorption maxima at pH 2 and increased with the increase in temperature. The changes in pH and in the ratio of choride ions released to chromate ions sorbed suggested that the anion exchange process was accomanied by hydrolysis and Donnan invasion. The FTIR spectra and isoeteric heat of sorption besides showing the hydrolysis / Donnan invasion of resin under different experimental conditions of concentration, pH and temperature also showed the dimerization of the chromate species inside the resin phase.

Adsorption↗

Immunological detection of DNA-protein complexes induced by chromate.

A select group of non-histone proteins becomes complexed to DNA after Chinese hamster ovary (CHO) cells are treated with potassium chromate. The most abundant complexed protein has a mol. wt of approximately 45 kd and is thought to be actin. An antiserum to the chromate-induced DNA-protein complexes (DPCs) was prepared to facilitate the study of these complexes. Rather than detecting the predominant silver-stained proteins of DPCs described in an earlier study, this antiserum reacts primarily with an acidic 95-kd protein (p95) that does not silver stain. The antiserum can be used routinely to assay for the induction of p95-DNA complexes produced by chromate and perhaps by other carcinogens. Immunofluorescent staining of CHO cells and immunoblotting of cell fractions show the reactive antigens are within the cell nucleus. Blotting experiments with the antiserum indicate the chromate-induced p95-DNA complex dissociates in the presence of 2-mercaptoethanol, suggesting similarity to the DPCs formed by carcinogenic platinum compounds. A reduced species of chromium (probably Cr3+) may form DPCs by binding to the nitrogen, oxygen or sulfur atoms of proteins and DNA. These results illustrate the usefulness of immunological detection methods to study specific DNA-protein interactions induced by carcinogens. The possible relevance of DPCs in the carcinogenic process is discussed.

Animals↗

Effect of in vivo chromate, acetone and combined treatment on rat liver in vitro microsomal chromium(VI) reductive activity and on cytochrome P450 expression.

Cytochrome P450 (P450IIE1) in rat has previously been shown to exhibit high chromate [Cr(VI)] reductase activity (Mikalsen et al. 1991). The present study reports on the effect of chromate treatment in vivo in rats and on the modulating effect of acetone + fasting on chromium(VI) toxicity. No effect of intraperitoneal injection with 5 mg chromate/kg was observed, whereas 15 mg chromate/kg decreased the liver microsomal Cr(VI) reductase activity by about 30% in in vitro microsomal incubations. In addition, the P450 and cytochrome b5 contents were decreased by about 30% and 25% respectively. Acetone + fasting caused increases of total microsomal P450 and cytochrome b5 contents, associated with similar increases in apoproteins P450IIE1 and P450IIB1 + 2, and their corresponding mRNA, and apoprotein NADPH-P450 reductase, as well as NADPH-P450 reductase and microsomal Cr(VI) reductive activities. Related to acetone + fasting alone, when given in combination with chromate (15 mg/kg) the Cr(VI) reductive activity was decreased by about 30%, associated with decreases in the P450 and cytochrome b5 contents, 65% and 35% respectively. This further reduced the apoprotein levels of P450IIB1 + 2, P450IIE1, and NADPH-P450 reductase to 90%, 60%, and 40%, respectively, and the mRNA levels of P450IIB2 and P450IIE1. No effect was observed on NADPH-P450 reductase activity. This dose also caused some macroscopic alterations in the liver. In contrast, the P450IIE1 apoprotein level in the lung was apparently stabilized or even increased by chromate in rats treated with acetone + fasting.(ABSTRACT TRUNCATED AT 250 WORDS)

Acetone↗

Chromate resistance plasmid in Pseudomonas fluorescens.

Chromate resistance of Pseudomonas fluorescens LB300, isolated from chromium-contaminated sediment in the upper Hudson River, was found to be plasmid specified. Loss of the plasmid (pLHB1) by spontaneous segregation or mitomycin C curing resulted in a simultaneous loss of chromate resistance. Subsequent transformation of such strains with purified pLHB1 plasmid DNA resulted in a simultaneous re-acquisition of the chromate resistance phenotype and the plasmid. When pLHB1 was transferred by conjugation to Escherichia coli, the plasmid still conferred chromate resistance.

Anti-Bacterial Agents↗

A cohort study of bronchial carcinomas in workers producing chromate pigments.

A cohort study of the incidence of bronchial cancer in male workers in a small company producing chromate pigments is presented. Altogether 133 workers had been employed by the company from the time production was started in 1948 until the end of 1972. Workers with more than three years employment were included in the study, and three cases of bronchial carcinoma were found among the 24 workers who fulfilled this requirement. Based on the data of the Cancer Registry of Norway the risk of bronchial cancer for a corresponding group of the general population was found to be 0.079, which gives a risk ratio for exposed workers of approximately 38. The average age of the cancer patients was as low as 50 years at the time of diagnosis. All workers in the company had been exposed mainly to zinc chromate dust, and the exposure levels of the workers developing bronchial cancers had probably been from 0-5 to 1-5 mg Cr/m-3 for six to nine years. Two of the three patients were smokers. It is assumed that exposure to chromate pigments, and probably to zinc chromate, may be related to the increased incidence of bronchial cancer in this group of workers. The possibility of a contributing effect of tobacco smoking in at least two of the three cases cannot be ruled out.

Adult↗

Insulin-like action of chromate on glucose transport in isolated rat adipocytes.

The effects of chromium compounds on 3-O-methylglucose (3-O-MG) transport were studied in isolated rat adipocytes. Sodium chromate significantly stimulated 3-O-MG uptake into adipocytes in a dose-dependent manner without altering the equilibrium space of 3-O-MG in adipocytes. The stimulatory effect reached the maximum at 300 microM, and the effect was 60-70% of the maximal insulin effect that was obtained with 20 nM insulin. The chromate concentration achieving a half-maximal effect was estimated at 50 microM. The effect of the combination of 1 mM chromate and 20 nM insulin was equipotent to that of 20 nM insulin alone, which showed that these two effects were not additive. The stimulatory effects of 1 mM chromate and 20 nM insulin were entirely abolished in adipocytes deprived of ATP, which indicated that these effects were completely ATP-dependent. Judging from experiments using various chromium compounds, CrO4(2-) was responsible for the insulinomimetic action. These results indicate that the action of CrO4(2-) is exerted through a mechanism analogous to that of insulin action, and that CrO4(2-) is a novel and useful tool for studying issues involved in insulin actions.

3-O-Methylglucose↗

DNA-protein complexes induced by chromate and other carcinogens.

DNA-protein complexes induced in intact Chinese hamster ovary cells by chromate have been isolated, analyzed, and compared with those induced by cis-platinum, ultraviolet light, and formaldehyde. Actin has been identified as one of the major proteins complexed to DNA by chromate based upon its molecular weight, isoelectric point, positive reaction with an actin polyclonal antibody, and proteolytic mapping. Chromate and cis-platinum both complex proteins of similar molecular weight and isoelectric points, and these complexes can be disrupted by chelating agents and sulfhydryl reducing agents, suggesting that the metal itself is participating in binding rather than having a catalytic or indirect role (i.e., oxygen radicals). In contrast, formaldehyde complexed histones to the DNA, and these complexes were not disrupted by chelating or reducing agents. An antiserum raised to chromate-induced DNA-protein complexes reacted primarily with 97,000 kDa protein that did not silver stain. Slot blots, as well as Western blots, were used to detect formation of p97 DNA crosslinks. This protein was complexed to the DNA by all four agents studied.

Animals↗

Generation of hydroxyl radical by chromate in biologically relevant systems: role of Cr(V) complexes versus tetraperoxochromate(V).

While Cr(V) species and .OH radicals have been suggested to play significant roles in the mechanism of chromate-related carcinogenesis, controversy still exists regarding the identity of the Cr(V) species and their role in the generation of .OH radicals. Some recent studies have suggested that the primary Cr(V) species involved is the tetraperoxochromate(V) (CrO8(3-)) ion, which produces .OH radical either on decomposition or by reaction with H2O2. The present study utilized ESR and spin trapping techniques to probe this mechanism. The results obtained show that (i) CrO8(3-) is not formed in any significant quantity in the reaction of chromate with biologically relevant reductants such as glutathione, glutathione reductase, NAD(P)H, ascorbate, vitamin B2, etc. (ii) Decomposition of CrO8(3-), or its reaction with H2O2 does not generate any significant amount of .OH radicals. (iii) The major Cr(V) species formed are complexes of Cr(V) with reductant moieties as ligands. (iv) These Cr(V) complexes generate .OH radicals from H2O2 via Fenton-like reaction. The present study thus disagrees with the recently proposed "tetraperoxochromate(V) theory of carcinogenesis from chromate." Instead, it suggests an alternative mechanism, which might be labeled as "the Cr(V)-complexation-Fenton reaction model of carcinogenesis from chromate.

Ascorbic Acid↗

Chemical models important in understanding the ways in which chromate can damage DNA.

Chromate is an established human carcinogen. There have been many studies of the reactivity of chromate aimed at improving understanding of chromate toxicity. In the present paper a number of conclusions of these studies are reviewed and considered in the light of new results obtained in our laboratories. A number of hypotheses are considered; it is concluded, however, that it is impossible to reconcile the generation of strand breaks by chromate during its reduction by glutathione with any simple mechanism involving the generation of DNA lesions by free hydroxyl radicals. Kinetic, spin-trapping, and competition kinetic studies, based on a strand-breaking assay, are reported in support of this conclusion.

Animals↗

Chromate removal by an iron sorbent: mechanism and modeling.

A solution containing chromate was treated using waste shot-blast fines recovered from surface finishing operations in a cast-iron foundry as a sorbent in batch and fixed-bed modes. Equilibrium experiments for initial chromate concentrations of 5 to 10 ppm produced a pH-adsorption edge that exhibits removal of chromium (Cr) over a broad pH range, with adsorption capacities that compare favorably to those reported for other adsorbents such as activated carbon and commercial iron oxides. Surface complexation modeling of adsorption equilibria suggests the formation of monodentate, inner-sphere complexes with chromate (CrO4(2-)) and bichromate (HCrO4(-)). Adsorption of Cr(VI) at iron oxy-hydroxide sites appears to be the primary mechanism of chromium removal at neutral pH. At lower pH values (for example, pH 4), reduction to Cr(III) is assumed to contribute to the increasing removal as a function of decrease in pH. There is also evidence to support the formation of Cr(III)-iron (Fe)(III) coprecipitate following Cr(VI) reduction by dissolved Fe(II). Using equilibrium constants for the two surface complexation reactions evaluated from a triple-layer model description of the oxide-water interface, chromate removal in a short fixed bed of fines was simulated using a dual mass-transfer kinetic model. Rate coefficients determined from model calibration of the short column were used to predict experimental breakthrough curves in columns with empty bed contact times (EBCTs) up to four times the short column. For an influent chromium concentration and pH of 5 ppm and 7.0, respectively, a solid-phase loading capacity of 9.5 +/- 0.3 mg/g was achieved at exhaustion. Predictive model runs indicate that, for this case, an EBCT of 2.0 to 2.5 minutes is optimum for achieving a target effluent concentration of less than or equal to 0.05 mg/L chromium as Cr(VI).

Adsorption↗

Purification and partial characterization of a chromate reductase from Bacillus.

A soluble NADH-dependent enzyme capable of reducing hexavalent chromium [Cr(VI)] to the trivalent form [Cr(III)] was purified from chromate-resistant Bacillus QC1-2. An enriched single protein band of 24 kDa was observed by SDS-PAGE following HPLC ion-exchange and size-exclusion procedures. In the latter step, the chromate reductase showed a molecular mass of 44 kDa, which suggested that the enzyme consists of two subunits of about 24 kDa. Purified chromate reductase displayed optimal activity at a temperature and pH of 37 degrees C and 7.0, respectively. The enzyme showed a Km of 0.35 mM for chromate and a Vmax of 50 nmol Cr(VI) reduced per minute per mg protein.

Bacillus↗

Chromate nephrotoxicity in developing rats. Significance of Cr(VI) reduction in rat kidney tissue.

Chromate reduction was studied in 9000 x g supernatant of renal tissue. It was shown that GSH plays an important role. Additionally, heat-sensitive factor(s) seem to be involved. Age differences in chromate nephrotoxicity parallel differences in chromate reduction in vitro. The lower reductive activity of young rats can be raised to adult values by GSH-addition. Fasting reduces GSH concentration, as well as chromate nephrotoxicity, in vivo.

Aging↗

Hypersensitivity to chromated catgut sutures: a case report and review of the literature.

Chromated catgut sutures are used extensively in general surgical procedures. Although delayed hypersensitivity reactions to this material are difficult to diagnose post-operatively--ie, they may be interpreted as non-specific complications of surgery--the condition can be readily evaluated by patch testing with chromate or intracutaneous placement of the actual suture material. Multiple reports in the literature suggested that there was an increased frequency of post-operative adhesions, wound dehiscence, infection, and even intestinal obstruction along the line of suture material to which the patient had been sensitized during a previous surgery. Particularly with the widespread availability of fully synthetic suture materials, which appear to be less sensitizing, an increased awareness of chromate hypersensitivity in patients undergoing repeated surgical procedures may decrease the frequency of subsequent inflammatory post-operative complications. We report a patient with a history of multiple abdominal surgical procedures, eosinophilic cystitis surrounding the suture material, prolonged post-operative pain, and inflammatory masses at the sites of previous surgery associated with a positive delayed hypersensitivity skin reaction to patch test chromate and to intradermal chromic catgut.

Abdomen↗

The carcinogen chromate induces DNA cross-links in rat liver and kidney.

DNA cross-links were found in nuclei isolated from the liver and kidney of rats treated with chromate. A dose-dependent relationship between chromate exposure and total DNA cross-links was determined using the alkaline elution technique. DNA cross-links in kidney were mainly DNA-protein in nature. Chromate also induced a small amount of interstrand DNA cross-links in kidney. Liver nuclei contained protein-associated DNA single strand breaks in addition to DNA-protein cross-links. The organotropic DNA damage induced by chromate is discussed in relation to the carcinogenicity and toxicity of chromium(VI) compounds.

Animals↗

The eye of the blue acara (Aequidens pulcher, Cichlidae) grows to compensate for defocus due to chromatic aberration.

By rearing fish in various monochromatic illuminations we investigated (1) the potential for compensation of refractive error due to chromatic aberration, (2) the contributions of the chromatic channels to emmetropization, and (3) the role of color cues in the control of eye growth. Cichlid fish (Aequidens pulcher) were reared for 6 months (12 h light/12 h dark) in monochromatic lights (623.5, 534.1, 485.0 nm; spectral purity 5-10 nm). Light levels were isoirradiant at 1.1.10(12) quanta/s/cm2. Two control groups were reared in white light with down-welling illuminances of 0.2 and 33 lx. Nasotemporal diameters (NTDs) of the eyes were measured in relation to lens size. Due to the oblique axis of highest acuity vision in cichlids, NTD is considered to be a more important dimension than axial length. Variances in NTD were equally small in all rearing groups. NTDs were enlarged with increasing wavelengths of the rearing lights with highly significant values over controls in the red-light group. The wavelength-dependent size of the eyes matched the changes in focal length due to longitudinal chromatic aberration. Complete recovery from eye enlargement was observed after fish reared in red light were exposed to a white light regime for 5 weeks. Small variances in NTD in all groups indicated stringent control of eye growth in the absence of color cues. The reversibility of the increase in NTD in fish reared in red light suggests that the eyes were emmetropized by visually guided mechanisms. Eye size in fish reared in white light was intermediate between the values expected if only blue-sensitive single or the red- and green-sensitive double cones contributed to the control of eye growth. This suggests that all chromatic channels participate in emmetropizing the fish eye.

Adaptation, Ocular↗

Human axial chromatic aberration found not to decline with age.

Millodot (1976) reported a dramatic decline in the amount of axial chromatic aberration of the human eye with age. The present study represents a failure to replicate that finding using a more standard procedure. No difference in chromatic aberration was found between a young and an older group of observers. Also, the chromatic aberrations of two observers which had been measured 25 years previously showed no decline when these measurements were repeated, even though their ages at first and second testing straddled the period over which Millodot reported the most change in chromatic aberration.

Adult↗