vitamin-k-semiquinone-radical and quinone

We synthesized novel vitamin K derivatives by converting the naphthoquinone group to benzene derivatives and benzoquinone. We evaluated their neuronal differentiation activities to investigate the effect of the quinone moiety on this process. We observed that the 1,4-quinone as well as the side chain part play important roles in neuronal differentiation. We also performed QSAR analysis to predict the compounds which would have higher differentiation activity.

Topics: Animals; Benzene Derivatives; Benzoquinones; Cell Differentiation; Dose-Response Relationship, Drug; Mice; Molecular Structure; Naphthoquinones; Neurons; Quantitative Structure-Activity Relationship; Vitamin K

Lactic acid bacteria were examined for their ability to produce quinone compounds, which may include dietary sources of menaquinones. Isoprenyl quinones in bacterial cells grown in a synthetic medium were extracted and analyzed by thin layer chromatography. Lactococcus lactis ssp. cremoris (three strains), Lactococcus lactis ssp. lactis (two strains), and Leuconostoc lactis were selected as high producers of quinone that synthesized more than 230 nmol of quinones/g of dried cells. The quinones were presumed to be menaquinone-7 to -10 by high performance liquid chromatography. Precise molecular weights were determined by mass spectrometry for Lactococcus lactis ssp. cremoris YIT 2011 and Leuconostoc lactis YIT 3001 and identified as menaquinone-8 and -9 for the former and menaquinone-9 and -10 for the latter. Those strains, when grown either in reconstituted nonfat dry milk or a soymilk medium, produced a beneficial quantity for dietary supplement (i.e., 29 to 123 micrograms of menaquinones/L of the fermented medium).

Topics: Benzoquinones; Chromatography, High Pressure Liquid; Chromatography, Thin Layer; Glycine max; Lactococcus lactis; Leuconostoc; Mass Spectrometry; Molecular Weight; Vitamin K

Our previous studies demonstrated that menadione is cytotoxic to rat platelets. In an attempt to assess the relative contributions of enzymatic redox cycling versus arylation in menadione-induced cytotoxicity, we have studied three quinones with different mechanisms of action: 2,3-dimethoxy-1,4-naphthoquinone (DMNQ; pure redox cycler), menadione (both redox cycler and arylator), and 1,4-benzoquinone (BQ; pure arylator). BQ was more toxic to rat platelets than menadione, while DMNQ did not cause LDH leakage at all. Cellular uptake kinetics revealed that DMNQ concentration taken up by the cells was equivalent to that decreased in incubation medium. On the other hand, the concentrations of BQ and menadione taken into the cells were significantly lower than the decreases in concentrations seen in the incubation medium. This suggests indirectly that BQ and menadione may have undergone arylation, binding to glutathione (GSH) or protein thiols. The difference in arylation capacity between BQ and menadione was well correlated with their relative cytotoxicity (LDH leakage) observed in platelets. All three quinones caused a rapid, extensive depletion of intracellular GSH in platelets. Treatments with BQ and menadione did not result in formation of oxidized glutathione (GSSG), whereas DMNQ showed a time-dependent increase in GSSG. Altogether, these results suggest that enzymatic redox cycling does not play a critical role in quinone-induced cytotoxicity in rat platelets, while arylation is likely to be quinone's primary mechanism of action.

Topics: Alkylation; Animals; Benzoquinones; Blood Platelets; Cell Survival; Chromatography, High Pressure Liquid; Female; Glutathione; Homeostasis; L-Lactate Dehydrogenase; Naphthoquinones; Oxidative Stress; Oxygen Consumption; Quinones; Rats; Rats, Sprague-Dawley; Vitamin K

Quinones caused quenching of Chl a fluorescence in native and model systems. Menadione quenched twofold the fluorescence of Chl a and BChl a in pea chloroplasts, chromatophores of purple bacteria, and liposomes at concentrations of 50-80 microM. To obtain twofold quenching in Triton X-100 micelles and in ethanol, the addition of 1.3 mM and 11 mM menadione was required, respectively. A proportional decrease in the lifetime and yield of Chl a fluorescence in chloroplasts, observed as the menadione concentration increased, is indicative of the efficient excitation energy transfer from bulk Chl to menadione. The decrease in the lifetime and yield of fluorescence was close to proportional in liposomes, but not in detergent micelles. The insensitivity of the menadione quenching effect to DCMU in chloroplasts, and similarity of its action in chloroplasts and liposomes indicate that menadione in chloroplasts interacts with antenna Chl, i.e., nonphotochemical quenching of fluorescence occurs.

Topics: Bacterial Chromatophores; Bacteriochlorophylls; Benzoquinones; Chlorophyll; Chlorophyll A; Chloroplasts; Diuron; Fluorescence; Liposomes; Micelles; Pisum sativum; Quinones; Rhodobacter sphaeroides; Rhodospirillum rubrum; Spectrometry, Fluorescence; Ubiquinone; Vitamin K

We report a simple fluorometric method for the continuous monitoring of mitochondrial membrane potential and cell viability in suspensions of hepatocytes exposed in vitro to cytotoxic agents. Suspensions of freshly isolated hepatocytes (10(6) cells/mL) preloaded with rhodamine 123 (Rh 123, 100 mumol/L) are transferred to a thermostatically controlled mixed cuvette to which the desired cytotoxic agent is added. Rh 123 is a cationic fluorophore that is actively accumulated by cells in direct proportion to the mitochondrial membrane potential. Cell viability was estimated by monitoring propidium iodide (PI) fluorescence. Exposure of cell suspensions to the mitochondrial uncoupling agent FCCP caused an immediate and titratable increase in Rh 123 fluorescence. Subsequent treatment with digitonin did not change Rh 123 fluorescence, suggeseting that Rh 123 equilibrates rapidly across the intact cell membrane. Likewise, treatment of hepatocyte suspensions with inhibitors of mitochondrial respiration (rotenone, cyanide, or menadione) caused an immediate increase in Rh 123 fluorescence. This was accompanied by a progressive increase in PI fluorescence, suggesting a causal relationship between mitochondrial depolarization and cell injury. In contrast, 1,4-benzoquinone caused a time-dependent and linear increase in PI fluorescence that paralleled changes in Rh 123 fluorescence. Comparing the time courses for changes in PI and Rh 123 fluorescence suggests that for benzoquinone, the depolarization of the mitochondria is a consequence rather than a cause of the cell injury. This modified procedure provides a simple and specific technique for continuously monitoring mitochondrial membrane potential and cell viability in suspensions of freshly isolated hepatocytes. The advantage is that there is no need to separate cells from the incubation medium, making it possible to record real-time changes in mitochondrial membrane potential and cell viability throughout the in vitro exposure period.

Topics: Animals; Benzoquinones; Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone; Cell Survival; Cyanides; Fluorescent Dyes; Fluorometry; Indicators and Reagents; Liver; Male; Membrane Potentials; Mitochondria, Liver; Propidium; Rats; Rats, Sprague-Dawley; Rhodamine 123; Rhodamines; Rotenone; Transfection; Uncoupling Agents; Vitamin K

Menadione (2-methyl-1,4-naphthoquinone, a redox cycling and arylating quinone; 5-100 microM) inhibited the canalicular vacuolar accumulation (CVA) of a fluorescent bile acid, cholyl-lysyl-fluorescein (CLF), in rat hepatocyte couplets. This was associated with depletion of reduced glutathione and accumulation of oxidized glutathione, the latter indicating that the concentrations of menadione used were able to induce oxidative stress. There was no associated cytotoxicity as indicated by ATP content. Treatment of couplets with the redox cycling quinone 2,3-dimethoxy-1,4-naphthoquinone (up to 100 microM) had relatively little effect on CVA, suggesting that the magnitude of reactive oxygen formation induced by this compound was insufficient to disrupt canalicular integrity. In comparison, the arylation of protein thiol groups by p-benzoquinone (up to 100 microM) proved to be more potent in inhibiting canalicular vacuolar accumulation. The predominant mechanism of menadione-induced inhibition of couplet hepatobiliary function is therefore more likely to involve the arylation of critical thiol groups (such as those in the F-actin cytoskeleton) rather than their oxidation. The oxidative effects of menadione could, however, potentiate the deleterious effects induced by arylation, such as by reduced glutathione depletion.

Topics: Adenosine Triphosphate; Animals; Benzoquinones; Biliary Tract; Cell Membrane; Cell Separation; Glutathione; Homeostasis; Liver; Naphthoquinones; Oxidation-Reduction; Rats; Vacuoles; Vitamin K

1. The effect of a redox cycler and arylator (menadione) and a pure arylator quinone (benzoquinone) was studied on different NADPH generating and consuming processes in isolated mouse hepatocytes. 2. Menadione inhibited gluconeogenesis from alanine but not from fructose or glycerol. 3. Drug oxidation measured as aniline hydroxylation and aminopyrine N-demethylation could be inhibited by menadione in microsomal membrane and in isolated hepatocytes both from fed or fasted animals. 4. Ureogenesis in isolated hepatocytes from fed mice could not be inhibited even by high concentration of menadione, while in cells from fasted animals menadione was inhibitory at high concentration in the presence of gluconeogenic precursor and at lower concentration in the absence of it. 5. Benzoquinone did not inhibit the above mentioned processes.

Topics: Aniline Hydroxylase; Animals; Benzoquinones; Cells, Cultured; Gluconeogenesis; Liver; Male; Mice; NADP; Oxidation-Reduction; Urea; Vitamin K

The effects of quinones (benzoquinone, menadione, and doxorubicin) on the superoxide production in cell free systems (xanthine oxidase and rat liver microsomes) and of polycationic electrolyte- and latex-stimulated rat peritoneal macrophages have been studied. Contradictory results were obtained in cell free systems when two traditional assays for detection of superoxide ion, the cytochrome c reduction and the lucigenin-dependent chemiluminescence (CL), were used: all quinones inhibited the lucigenin-dependent CL at sufficiently large concentrations, but they did not inhibit at all the reduction of cytochrome c. It was proposed that the cytochrome c assay gave erroneous results due to the reversibility of the interaction of semiquinones with dioxygen. The effect of quinones on the superoxide production by peritoneal macrophages was biphasic: all quinones stimulated the O2-. formation at low concentrations and inhibited it at elevated concentrations. It was concluded that among the quinones studied, only menadione was capable of stimulating the superoxide production via a one-electron transfer mechanism in cell free systems, while the stimulatory effect of small concentrations of quinones on the O2-. production in macrophages was possibly due to their action on the activation of NADPH oxidase.

Topics: Animals; Benzoquinones; Cell-Free System; Cytochrome c Group; Doxorubicin; Luminescent Measurements; Macrophages; Microsomes, Liver; Nitroblue Tetrazolium; Oxidation-Reduction; Quinones; Rats; Superoxides; Vitamin K; Xanthine Oxidase; Xanthines

The nucleophilic addition of GSH to quinonoid compounds, characterized as a 1,4-reductive addition of the Michael type, was studied with p-benzoquinone- and 1,4-naphthoquinone epoxides with different degree of methyl substitution. Identification and evaluation of molecular products from the above reaction were assessed by h.p.l.c. with either reductive or oxidative electrochemical detection, based on the redox properties retained in the molecular products formed. It was found that the degree of methyl substitution of the quinone epoxide, from either the 1,4-naphthoquinone- or p-benzoquinone epoxide series, determined their rate of reaction with GSH. The reductive addition implied the rearrangement of the quinone structure with opening of the epoxide ring yielding as the primary product a hydroxy-glutathionyl substituted adduct of either p-benzohydroquinone or 1,4-naphthohydroquinone. The primary product undergoes elimination reactions and redox transitions which bring about a number of secondary molecular products. The distribution pattern of the latter depends on the degree of methyl substitution of the quinone epoxide studied and on the concentration of O2 in the solution. The occurrence of the hydroxy-substituent in position alpha, adjacent to the carbonyl group, enhances the autoxidation properties of the compound resulting in an augmented O2 consumption and H2O2 production. Therefore, it could be expected that the chemical reactivity of the products originating from the thiol-mediated nucleophilic addition to quinone epoxides would be of toxicological interest.

Topics: Benzoquinones; Chemical Phenomena; Chemistry; Chromatography, High Pressure Liquid; Epoxy Compounds; Ethers, Cyclic; Free Radicals; Glutathione; Hydrogen Peroxide; Kinetics; Naphthoquinones; Oxidation-Reduction; Oxygen; Quinones; Vitamin K

Quinones may be toxic by a number of mechanisms, including oxidative stress caused by redox cycling and arylation. This study has compared the cytotoxicity of four quinones, with differing abilities to arylate cellular nucleophiles and redox cycle, in relation to their effects on cellular pyridine nucleotides and ATP levels in rat hepatocytes. Non-toxic concentrations (50 microM) of menadione (redox cycles and arylates), 2-hydroxy-1,4-naphthoquinone (neither arylates nor redox cycles via a one electron reduction) and 2,3-dimethoxy-1,4-naphthoquinone (a pure redox cycler) all caused markedly similar changes in cellular pyridine nucleotides. An initial decrease in NAD+ was accompanied by a small, transient increase in NADP+ and followed by a larger, prolonged increased in NADPH and total NADP+ + NADPH. At toxic concentrations (200 microM), the quinones caused an extensive depletion of NAD(H), an increase in levels of NADP+ and an initial rise in total NADP+ + NADPH, prior to a decrease in ATP levels and cell death. Nucleotide changes were not observed with non-toxic (20 microM) or toxic (100 microM) concentrations of p-benzoquinone (a pure arylator) and ATP loss accompanied or followed cell death. A novel mechanism for the activation of 2-hydroxy-1,4-naphthoquinone has been implicated. Our findings also suggest that a primary event in the response of the cell to redox cycling quinones is to bring about an interconversion of pyridine nucleotides, possibly mediated by an NAD+ reduction, in an attempt to combat the effects of oxidative stress.

Topics: Animals; Benzoquinones; Cell Survival; In Vitro Techniques; Liver; Male; NAD; NADP; Naphthoquinones; Oxidation-Reduction; Quinones; Rats; Rats, Inbred Strains; Vitamin K

Topics: Antifibrinolytic Agents; Benzoquinones; Blood Coagulation; Chemical Phenomena; Chemistry; Naphthoquinones; Pharmacology; Quinones; Rabbits; Research; Vitamin K

Topics: Antifibrinolytic Agents; Benzoquinones; Blood Coagulation; Humans; Naphthoquinones; Quinones; Ubiquinone; Vitamin K

Topics: Amines; Benzoquinones; Blood Pressure; Blood Pressure Determination; Hypertension; Quinones; Vitamin K; Vitamins