Chromium accumulation by living yeast at various environmental conditions

Yeast tolerance to Cr (III) and Cr (VI) as well as chromium accumulation potential were shown to depend on treatment time, metal concentration, biomass density and the phase of growth. Kinetic studies as exemplified by Pichia guilliermondii ATCC 201911 revealed a biphasic mode of Cr (III) uptake: a rapid sorption phase was followed by a slow process of accumulation, in which the contribution of the cell-bound Cr fraction increased, while the total cellular Cr level remained constant. Cr (VI) uptake was characterized by a time-dependent increase of total Cr and by a constant fractional contribution of the cell-adsorbed chromium, which suggests that the amount of cell-accumulated Cr also tended to increase over time. The resistance to Cr and metal accumulation levels were substantially elevated for a given strain when cultures were treated at high initial biomass densities (1 mg dry weight/ml) of exponentially proliferating cells. Maximum accumulation capabilities ranged between 4.0 and 13 mg Cr (III)/g dry weight and 2-6.7 mg Cr (VI)/g dry weight. The total cell-accumulated Cr contained 29.3% and 52.3% of organically bound chromium for the treatment of P. guilliermondii with Cr (III) and Cr (VI), respectively. Selected yeast strains, under specified physiological conditions, can be applied for bioremediation of environmental Cr contamination, and might be useful too for attempts to obtain chromium-enriched biomass containing biostabilized and nontoxic Cr forms for nutritional applications.     

Mitochondrial toxicity and antioxidant activity of a prenylated flavonoid isolated from Dalea elegans

The prenylated flavanone 2'-4'-dihidroxy-5'-(1" '-dimethylallyl)-6-prenylpinocembrin) (6PP), isolated from the roots of Dalea elegans, shows antimicrobial activity. The aim of this study was to evaluate mitochondrial toxicity and antioxidant properties of 6PP. Addition of micromolar concentrations of 6PP to rat liver mitochondria, stimulated O2 uptake in state 4 and inhibited it in state 3 when malate-glutamate was the respiratory substrate, and inhibited O2 uptake in state 3 when succinate was the substrate. Highest concentration of 6PP also inhibited O2 uptake in state 4 in the latter case; in both conditions, respiratory control index values were decreased. This flavanone collapsed the mitochondrial membrane potential in a concentration-dependent manner. 6PP also inhibited F0F1-ATPase activity in coupled mitochondria and in submitochondrial particles. In the latter, this compound also inhibited NADH oxidase and succinate dehydrogenase activities. HEp-2 cells were incubated for 24 h with 6PP in presence or absence of 0.5% albumin. As measured by reduction of the mitochondrial-related probe MTT, in the albumin-free condition, 6PP was cytotoxic in a concentration-dependent manner; on the other hand, albumin decreased 6PP effect. In addition, in rat liver microsomes 6PP: (1) inhibited the enzymatic lipid peroxidation, (2) exhibited significant scavenging activity, measured by DPPH reduction assay and (3) demonstrated significant antioxidant activity by decreasing the reduction of Mo(VI) to Mo(V). We suggest that 6PP impairs the hepatic energy metabolism by acting as mitochondrial uncoupler and by inhibiting enzymatic activities linked to the respiratory chain. 6PP also exerts both antioxidant and antiradical activities. Due to its cytotoxicity, this molecule, and its future structure developments, can be considered as a potentially promising therapeutic agent, for instance in cancer chemotherapy.     

Cytochrome c oxidase inhibition in the rice weevil Sitophilus oryzae (L.) by formate, the toxic metabolite of volatile alkyl formates

Volatile alkyl formates are potential replacements for the ozone-depleting fumigant, methyl bromide, as postharvest insecticides and here we have investigated their mode of insecticidal action. Firstly, a range of alkyl esters, ethanol and formic acid were tested in mortality bioassays with adults of the rice weevil, Sitophilus oryzae (L.) and the grain borer, Rhyzopertha dominica (F.) to determine whether the intact ester or one of its components was the toxic moiety. Volatile alkyl formates and formic acid caused similar levels of mortality (LC(50) 131-165 micromol l(-1)) to S. oryzae and were more potent than non-formate containing alkyl esters and ethanol (LC(50)>275 micromol l(-1)). The order of potency was the same in R. dominica. Ethyl formate was rapidly metabolised in vitro to formic acid when incubated with insect homogenates, presumably through the action of esterases. S. oryzae and R. dominica fumigated with a lethal dose of ethyl formate had eight and 17-fold higher concentrations of formic acid, respectively, in their bodies than untreated controls. When tested against isolated mitochondria from S. oryzae, alkyl esters, alcohols, acetate and propionate salts were not inhibitory towards cytochrome c oxidase (EC 1.9.3.1), but sodium cyanide and sodium formate were inhibitory with IC(50) values of 0.0015 mM and 59 mM, respectively. Volatile formate esters were more toxic than other alkyl esters, and this was found to be due, at least in part, to their hydrolysis to formic acid and its inhibition of cytochrome c oxidase.     

Additive effects and potential inhibitory mechanism of some common aromatic pollutants on in vitro mitochondrial respiration

The in vitro toxicity of multiple hydrophobic compounds was the focus of this study. A mitochondrial respiratory assay, sensitive to perturbations caused by hydrophobic chemicals, was utilized to measure the effects of individual aromatic hydrocarbon pollutants and their mixtures on mitochondrial respiratory function. Benzene, naphthalene, acenaphthene, and 1-chloronaphthalene, common industrial solvents shown to interact additively in vivo, were evaluated using this in vitro assay system. Mitochondrial respiration was inhibited 50% (EC₅₀) by 525 ppm (6.7 mM) benzene, 15 ppm (117 μM) naphthalene, 3.9 ppm (25.5 μM) acenaphthene, or 3.8 ppm (23.4 μM) 1-chloronaphthalene. NADH:O₂ oxidoreductase (NADH → O₂), NADH: ubiquinone oxidoreductase, and ubiquinol:O₂ oxidoreductase activities were inhibited by all four compounds, whereas succinate:O₂ oxidoreductase, cytochrome c oxidase, and duroquinol: O₂ oxidoreductase activities were not inhibited. Inhibition of mitochondrial respiration occurred at the level of ubiquinone (coenzyme Q₁₀) for all four aromatic hydrocarbons. The ultraviolet absorbance spectrum of isolated Q₁₀ was also altered by naphthalene, acenaphthene, or 1-chloronaphthalene, suggesting a specific interaction between that component of the respiratory chain and these aromatic hydrocarbons. Inhibition by a mixture of 2, 3, or 4 of the compounds tested was additive, reflecting a summation effect of each compound present in the mixture. This additive nature is consistent with previously reported effects of these compounds in vivo and with compounds having similar modes of action. The similar mode of action in vitro is a specific interaction with coenzyme Q₁₀, not a generalized membrane perturbation as speculated to occur in vivo, and is the likely mechanism for the observed additive toxicity.     

Comparison of acute absorption of commercially available chromium supplements

Chromium (Cr) supplements are available as picolinate, nicotinate or chloride (the latter primarily in multivitamin-mineral supplements). The picolinate form has been reported to be the best absorbed and most efficacious, but some reports question which form has superior absorption. The present study examined acute Cr absorption, based on 24h urinary Cr values, for picolinate, two types of nicotinate, and chloride in young adult, non-overweight females. College-aged women were given 200 microg of Cr as each of the four supplement types in random order accompanied by a small standardized meal, separated by at least a week washout. Cr picolinate produced significantly higher 24h urinary Cr than either of two nicotinate supplements or Cr chloride given in a multivitamin-mineral supplement. This difference was seen for absolute values of the urinary Cr and for percent increases. In conclusion, based on an indirect measure of acute absorption, Cr picolinate was superior to three other Cr complexes commonly sold as supplements.     

Interactions of chromium with microorganisms and plants

Chromium is a highly toxic non-essential metal for microorganisms and plants. Due to its widespread industrial use, chromium (Cr) has become a serious pollutant in diverse environmental settings. The hexavalent form of the metal, Cr(VI), is considered a more toxic species than the relatively innocuous and less mobile Cr(III) form. The presence of Cr in the environment has selected microbial and plant variants able to tolerate high levels of Cr compounds. The diverse Cr-resistance mechanisms displayed by microorganisms, and probably by plants, include biosorption, diminished accumulation, precipitation, reduction of Cr(VI) to Cr(III), and chromate efflux. Some of these systems have been proposed as potential biotechnological tools for the bioremediation of Cr pollution. In this review we summarize the interactions of bacteria, algae, fungi and plants with Cr and its compounds.     

微生物铬转化和抗性机制与生物修复研究进展

铬(Chromium,Cr)是过渡金属元素,在自然界中以六价[CrO_4~(2-),Cr_2O_7~(2-),Cr(Ⅵ)]和三价[Cr(OH)_3,Cr(Ⅲ)]为主。很多微生物在长期铬胁迫的条件下,进化出了一系列铬转化和抗性机制。微生物对铬的转化包括Cr(Ⅵ)的还原和Cr(Ⅲ)的氧化。微生物的Cr(Ⅵ)还原可以将毒性强的六价铬转化为毒性弱或无毒的三价铬,这类微生物有较强的土壤和水体铬污染治理潜力。Cr(Ⅲ)的氧化也在铬的生物地球化学循环过程中起着至关重要的作用。除了Cr(Ⅵ)的还原,微生物对铬的抗性机制还有:(1)减少摄入;(2)外排;(3)清除胞内氧化压力;(4)DNA修复。本文主要介绍微生物的铬转化和抗性机制,以及其在铬污染生物修复中应用的最新研究进展。     

Chromium uptake,retention and reduction in photosynthetic <Emphasis Type="Italic">Euglena gracilis</Emphasis>

Photosynthetic Euglena gracilis grown with different K₂CrO₄ concentrations was analyzed for its ability to take up, retain and reduce Cr(VI). For comparison, cells were also exposed to CrCl₃. Cellular Cr(VI) uptake at pH 7.2 showed a hyperbolic saturation pattern with K _m of 1.1 mM, V _m of 16 nmol (h × 10⁷ cells)⁻¹, and K _i sulfate of 0.4 mM. Kinetic parameters for sulfate uptake were similar, K _m = 0.83 mM, V _m = 15.9 nmol (h × 10⁷cells)⁻¹ and K _i chromate = 0.3 mM. The capacity to accumulate chromium depended on the ionic species, external concentration and pH of the incubation medium. Cr(VI) or Cr(III) accumulation was negligible in the acidic (pH 3.5) culture medium, in which Cr(VI) was abiotically reduced to Cr(III). At pH 7.2 Cr(VI) was fully stable and high accumulation (>170 nmol/1 × 10⁷ cells at 1 mM K₂CrO₄) was achieved; surprisingly, Cr(III) accumulation was also significant (>35 nmol/1 × 10⁷ cells at 1 mM CrCl₃). Cr(VI) was reduced by cells at pH 7.2, suggesting the presence of an external reductive activity. Cr(VI) induced an increased cysteine and glutathione content, but not in phytochelatins suggesting that chromium accumulation was mediated by monothiol compounds.     

Measurement of mitochondrial respiration in permeabilized murine neuroblastoma (N-2alpha) cells, a simple and rapid in situ assay to investigate mitochondrial toxins

Most mitochondria-based methods used to investigate toxins require the use of relatively large amounts of material and hence compromised sensitivity in assay. We adopted procedures from methods initially developed to diagnose mitochondrial encephalomyopathies and unified these into a single assay. Eukaryotic cell membranes are selectively permeabilized with digitonin to render a system in which mitochondrial respiration can be measured rapidly and with considerable sensitivity. Mitochondria remain intact, uninjured, and in their natural environment where mitochondrial respiration can be measured in situ under physiologically relevant conditions. This approach furthermore allows measurement of toxin effects on individual mitochondrial complexes. Numerous compounds at varying concentrations can be screened for mitochondrial toxicity, while the site of mitochondrial inhibition can be determined simultaneously. We used this assay to investigate, in murine neuroblastoma (N-2alpha) cells, the mitochondrial inhibitory properties of the parkinsonian-inducing proneurotoxin, 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), and its neurotoxic monoamine oxidase-B (MAO-B)-generated metabolite, the 1-methyl-4-phenylpyridinium species (MPP(+)). Within the time frame of each measurement (15 min), MPTP (< or = 1 mM) did not interfere with in situ mitochondrial respiration. As expected, MPP(+) was found to be a potent Complex I inhibitor but surprisingly also found to inhibit Complex IV. Optimized conditions for performing this assay are provided.     

Nutritional supplement chromium picolinate generates chromosomal aberrations and impedes progeny development in Drosophila melanogaster

Chromium picolinate, [Cr(pic)₃], is a popular nutritional supplement found in a variety of consumer products. Despite its popularity, safety concerns over its use have arisen. The supplement has been shown to generate clastogenic damage, mitochondrial damage, oxidative damage, and mutagenic effects in cultured cells and oxidative DNA damage and lipid peroxidation in rats. Recently [Cr(pic)₃] has been demonstrated to generate heritable genetic change and delays in progeny development in Drosophila melanogaster. Based on the damage to chromosomes of cultured cells and of animal models, similar chromosome damage appeared to be a likely source of the mutagenic effects of the supplement in Drosophila. The current three-part study examines the effects of several chromium-containing supplements and their components on hatching and eclosion rates and success of development of first generation progeny of adult Drosophila fed food containing these compounds. It further examines the effects of the compounds on longevity of virgin male and female adults. Finally, the chromosomes in the salivary glands of Drosophila late in the third instar larval stage, which were the progeny of Drosophila whose diets were supplemented with nutritional levels of [Cr(pic)₃], are shown to contain on average over one chromosomal aberration per two identifiable chromosomal arms. No aberrations were observed in chromosomes of progeny of untreated flies. The results suggest that human consumption of the supplement should be a matter of concern and continued investigation to provide insight into the requirements of chromium-containing supplements to give rise to genotoxic effects.     

Hypoglycemic potency of novel trivalent chromium in hyperglycemic insulin-deficient rats

Two sources of chromium III, "chromium 454" and "chromium picolinate," were tested in insulin-deficient Streptozocin-treated diabetic rats. This model was selected in order to evaluate the possibility of any hypoglycemic potency of chromium in a relative absence of blood insulin concentration. Three weeks of the treatment with CRC454 and CrP resulted in a 38% and 11% reduction of blood glucose levels, respectively. Body weight gains were equally improved by both treatments. Blood levels of CK, ALT and AST were significantly reduced by CRC454 and CrP. These results might suggest that any hypoglycemic effect of trivalent chromium under insulin-deficient conditions could be largely dependent upon the type of chromium agent and associated characteristics such as solubility and bioavalibility. In contrast, improvement of body weight gains and blood levels of CK, AST and ALT seems to be less dependent on the type of chromium compound under these experimental conditions. In conclusion, CRC454 showed significant reduction of hyperglycemia under insulin-deficient conditions.     

Effects of Pre- and Postnatal Exposure to Chromium Picolinate or Picolinic Acid on Neurological Development in CD-1 Mice

Chromium picolinate, Cr(pic)3, a popular dietary supplement marketed as an aid in fat loss and lean muscle gain, has also been suggested as a therapy for women with gestational diabetes. The current study investigated the effects of maternal exposure to Cr(pic)3 and picolinic acid during gestation and lactation on neurological development of the offspring. Mated female CD-1 mice were fed diets from implantation through weaning that were either untreated or that contained Cr(pic)3 (200 mg kg(-1) day(-1)) or picolinic acid (174 mg kg(-1) day(-1)). A comprehensive battery of postnatal tests was administered, including a modified Fox battery, straight-channel swim, open-field activity, and odor-discrimination tests. Pups exposed to picolinic acid tended to weigh less than either control or Cr(pic)3-exposed pups, although the differences were not significant. Offspring of picolinic acid-treated dams also appeared to display impaired learning ability, diminished olfactory orientation ability, and decreased forelimb grip strength, although the differences among the treatment groups were not significant. The results indicate that there were no significant effects on the offspring with regard to neurological development from supplementation of the dams with either Cr(pic)3 or picolinic acid.     

Catalytic conversion of glucose to 5-hydroxymethyl furfural using inexpensive co-catalysts and solvents

Efficient conversion of glucose to 5-hydroxymethyl furfural (5-HMF), a platform chemical for fuels and materials, was achieved using CrCl₂ or CrCl₃ as the catalysts with inexpensive co-catalysts and solvents including halide salts in dimethyl sulfoxide (DMSO) and several ionic liquids. 5-HMF (54.8%) yield was achieved with the CrCl₂/tetraethyl ammonium chloride system at mild reaction conditions (120 °C and 1 h). The 5-HMF formation reaction was found to be faster in ionic liquids than in the DMSO system. Effects of water in the reaction system, chromium valence and reaction temperature on the conversion of glucose into 5-HMF were discussed in this work.