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PubMed1988

Relative inhibitory potencies of flavonoids on 12-lipoxygenase of fish gill.

The relative efficacy of 10 flavonoid compounds and some common antioxidants in inhibiting 12-lipoxygenase of fish gill was determined. Lipoxygenase activity was measured by oxygen consumption with polarography and formation of hydroxy fatty acid using thin layer chromatography with autoradiography. Generally, the inhibition of 12-lipoxygenase by flavonoids was noncompetitive. The lipoxygenase product pattern did not change in the presence of flavonoids. Fisetin (3,3',4',7-tetrahydroxyflavone) and quercetin (3,3',4',5,7-pentahydroxyflavone) were the most potent inhibitors of 12-lipoxygenase, with IC50's of 0.25 and 0.4 microM respectively. These were compared with IC50's values of 5,1000 microM for BHA (t-butylhydroxyanisole) and BHT (t-butylhydroxytoluene) respectively. Possible inhibitory mechanisms and relationships between flavonoid structure and inhibitory potencies are discussed.

Animals↗

PubMed1991

Selenium in Kashin-Beck disease areas.

In this article, the duplication portion technique was used to determine the daily intakes of selenium and ten other elements in the 24-h total diets collected in the typical Kashin-Beck endemic areas, i.e., Shanxi Province and Inner Mongolia Autonomous of China. The contents of Ca, Mg, Cu, Fe, Zn, Mn, Al, Sr, Ba, and P in freeze-dried samples were determined by ICP-AES. Se was determined by differential pulse catalytic polarography. The average Se contents in total diets of Shanxi Kashin-Beck endemic and nonendemic areas were 0.009 and 0.021 micrograms/g (dry weight), respectively (P less than 0.001), corresponding to the daily intakes for Se of 4.6 and 10.5 micrograms. After the Se-supplemented fertilizer was applied (225 g of Na2SeO3/ha), the average Se content in total diets of Kashin-Beck disease area was increased to 0.0336 micrograms/g, which corresponded to the average daily intake for Se of 16.8 micrograms. In Inner Mongolia Kashin-Beck endemic and nonendemic areas, the average Se contents in total diets were 0.006 and 0.017 micrograms/g, respectively (p less than 0.001), corresponding to the average daily intakes for Se of 3 and 8.5 micrograms. The contents of other ten elements in total diets in endemic and nonendemic areas were reported and compared.

China↗

PubMed2000

Adenosine and amrinone reverse felypressin-induced depression of myocardial tissue oxygen tension in dogs.

PURPOSE: To determine whether continuous infusion of adenosine triphosphate (ATP), nitroglycerin (NTG) or amrinone (AM) would ameliorate the reductions in coronary blood flow (CBF) and myocardial oxygen tension (PmO2) induced by felypressin. METHODS: Seven open-chest dogs were studied under urethane and alpha-chloralose anesthesia. Hemodynamic variables including heart rate (HR), systolic blood pressure, diastolic blood pressure (DBP), mean pulmonary artery pressure, pulmonary capillary wedge pressure, CBF (ultrasound flowmetry), PmO2(polarography) and cardiac output (thermodilution method) were recorded. Felypressin was infused in a loading dose of 6 mIU x kg(-1) for five minutes and then continued at 0.2 mIU x kg(-1) x min(-1). After 30 min felypressin infusion, each agent was administered for 15 min to evaluate hemodynamic changes. Infusions were 100 and 200 microg x kg(-1) x min(-1) for ATP, 2.5 and 5 microg x kg(-1) x min(-1) for NTG, and 10 and 20 microg x kg(-1) x min(-1) for AM. RESULTS: After felypressin DBP increased by 17 +/- 5 (mean +/- SD) %; CBF decreased by 49 +/- 9%; CI decreased by 40 +/- 13%; HR decreased by 29 +/- 11%; PmO2 in the inner layer decreased by 21 +/- 7%. The Cl and CBF returned to baseline afterATP 100 and 200 microg x kg(-1) x min(-1), AM 10 and 20 microg x kg(-1) x min(-1), but not after NTG. The PmO2 in the inner layer returned to the baseline value by any infusion except for NTG 5 microg x kg(-1) min(-1) CONCLUSION: Adenosine and amrinone, but not nitroglycerin reverses the adverse cardiovascular effects of felypressin.

Adenosine↗

PubMed2003

A high carbohydrate diet does not induce hyperglycaemia in a mitochondrial glycerol-3-phosphate dehydrogenase-deficient mouse.

AIMS/HYPOTHESIS: The electrons of the glycolysis-derived reduced form of NADH are transferred to mitochondria through the NADH shuttle system. There are two NADH shuttles: the glycerol phosphate and malate-aspartate shuttle. Mice with a targeted disruption of mitochondrial glycerol-3-phosphate dehydrogenase, a rate-limiting enzyme of the glycerol phosphate shuttle, are not diabetic and have normal islet glucose-induced secretion. In this study, we analyzed if environmental factors, such as a high carbohydrate diet could contribute to the development of Type 2 diabetes mellitus in mice with a specific defective genetic background. METHODS: The mice were fed with a high carbohydrate diet for 1 and 6 months, and several biochemical parameters were analysed. The mitochondrial respiratory activity was assayed by polarography; and the islet function was studied by islet perifusion and pancreas perfusion. RESULTS: The high carbohydrate diet induced hyperglycaemia, hyperinsulinaemia, and islet hyperplasia in the wild-type and heterozygote mice. Activity of the respiratory chain complex I also increased in these mice. In contrast, these effects were not observed in the null mice fed with the diet; in addition, these null mice had an increased insulin sensitivity compared to wild-type mice. CONCLUSION/INTERPRETATION: The phenotype of the mice with an impairment of NADH shuttles does not worsen when fed a high carbohydrate diet; moreover, the diet does not compromise islet function.

Animals↗

PubMed1999

Protein kinase C effects on nerve function, perfusion, Na(+), K(+)-ATPase activity and glutathione content in diabetic rats.

AIMS/HYPOTHESIS: Increased protein kinase C activity has been linked to diabetic vascular complications in the retina and kidney, which were attenuated by protein kinase C antagonist treatment. Neuropathy has a vascular component, therefore, the aim was to assess whether treatment with WAY151 003 or chelerythrine, inhibitors of protein kinase C regulatory and catalytic domains respectively, could correct nerve blood flow, conduction velocity, Na(+),K(+)-ATPase, and glutathione deficits in diabetic rats. METHODS: Diabetes was induced by streptozotocin. Sciatic nerve conduction velocity was measured in vivo and sciatic endoneurial perfusion was monitored by microelectrode polarography and hydrogen clearance. Glutathione content and Na(+),K(+)-ATPase activity were measured in extracts from homogenised sciatic nerves. RESULTS: After 8 weeks of diabetes, sciatic blood flow was 50 % reduced. Two weeks of WAY151 003 (3 or 100 mg/kg) treatment completely corrected this deficit and chelerythrine dose-dependently improved nerve perfusion. The inhibitors dose-dependently corrected a 20 % diabetic motor conduction deficit, however, at high doses ( > 3.0 mg/kg WAY151003; > 0.1 mg/kg chelerythrine) conduction velocity was reduced towards the diabetic level. Sciatic Na(+),K(+)-ATPase activity, 42 % reduced by diabetes, was partially corrected by low but not high dose WAY151 003. In contrast, only a very high dose of chelerythrine partially restored Na(+),K(+)-ATPase activity. A 30 % diabetic deficit in sciatic glutathione content was unchanged by protein kinase C inhibition. The benefits of WAY151 003 on blood flow and conduction velocity were blocked by nitric oxide synthase inhibitor co-treatment. CONCLUSION/INTERPRETATION: Protein kinase C contributes to experimental diabetic neuropathy by a neurovascular mechanism rather than through Na(+),K(+)-ATPase defects.

Alkaloids↗

PubMed2001

Effects of an extracellular metal chelator on neurovascular function in diabetic rats.

AIMS/HYPOTHESIS: Increased oxidative stress has been causally linked to diabetic neurovascular complications, which are attenuated by antioxidants. There are several possible sources of reactive oxygen species in diabetes. Our aim was to assess the contribution of free radicals, produced by transition metal catalysed reactions, to early neuropathic changes. To this end, we examined, firstly, the effects of an extracellular high molecular weight chelator, hydroxyethyl starch-deferoxamine, which is expected to be confined to vascular space, on nerve perfusion and conduction deficits in diabetic rats and, secondly, the action of a single chelator dose. METHODS: Diabetes was induced by streptozotocin. In vivo measurements comprised sciatic nerve motor conduction velocity and endoneurial perfusion, monitored by hydrogen clearance microelectrode polarography. RESULTS: We found that 8 weeks of diabetes reduced sciatic blood flow and conduction velocity by 48.3 % and 19.9% respectively. Two weeks of intravenous treatment corrected these deficits. Starch vehicle was ineffective. The time-course of action of a single hydroxyethyl starch-deferoxamine injection was examined in diabetic rats. There was a rapid increase in nerve blood flow on day 1, which remained within the non-diabetic range for 9 days before declining to the diabetic level at day 27. In contrast, conduction velocity changes were slower, reaching the non-diabetic range at day 6 and declining to the diabetic level at day 27. CONCLUSION/INTERPRETATION: Extracellular transition metal catalysed reactions play a major role in the neurovascular deficits of experimental diabetes. Given the long-lasting effect of a single treatment, extracellular metal chelator therapy could be suitable for further assessment in clinical trials.

Animals↗

PubMed2001

Effect of the hydroxyl radical scavenger, dimethylthiourea, on peripheral nerve tissue perfusion, conduction velocity and nociception in experimental diabetes.

AIMS/HYPOTHESIS: Increased oxidative stress has been linked to diabetic neurovascular complications, which are reduced by antioxidants. Our aim was to assess the contribution of hydroxyl radicals to early neuropathic changes by examining the effects of treatment with the specific scavenger, dimethylthiourea, on nerve function and neural tissue blood flow in diabetic rats. METHODS: Diabetes was induced by streptozotocin. Measurements comprised sciatic nerve motor and saphenous nerve sensory conduction velocity. Responses to noxious mechanical and thermal stimuli were estimated by Randall-Sellito and Hargreaves tests respectively. Sciatic nerve and superior cervical ganglion blood flow were measured by hydrogen clearance microelectrode polarography. RESULTS: Eight weeks of diabetes reduced motor and sensory conduction velocity by 19.9% and 15.7% respectively, and these were completely corrected by 2 weeks of dimethylthiourea treatment. The ED50 for motor conduction was 9 mg kg(-1) x day(-1). Mechanical and thermal nociceptive sensitivities were 18.9% and 25.0% increased by diabetes, respectively, indicating hyperalgesia which was 70% reduced by dimethylthiourea. Sciatic endoneurial and superior cervical ganglion blood flows were 51.2% and 52.4% reduced by diabetes and there was an approximately 80% improvement with treatment. CONCLUSION/INTERPRETATION: Hydroxyl radicals seem to make a major contribution to neuropathy and vasculopathy in diabetic rats. Treatment with the hydroxyl scavenger, dimethylthiourea, was highly effective. The data suggest that the development of potent hydroxyl radical scavengers suitable for use in man could markedly enhance the potential therapeutic value of an antioxidant approach to the treatment of diabetic neuropathy and vascular disease.

Animals↗

PubMed2002

Electrocatalytic activity of cobalt phthalocyanine stabilized by different matrixes.

The behavior of cobalt phthalocyanine complexes incorporated inside an hydrotalcite-like clay (HT) or a sonogel-carbon composite has been investigated in order to develop chemically modified electrodes suitable for use as amperometric detectors. The electrocatalytic oxidation process of cysteine at this new electrode has been studied by cyclic voltammetry. For comparison, the oxidation of cysteine catalyzed by the cobalt phthalocyanine complex as a redox mediator, either dissolved in solution or entrapped inside the HT structure, has been followed by polarography. The sonogel-carbon composite electrode is stable and its response is repeatable. Cysteine oxidation is actually induced by the electrogenerated Co(III) complex, and the relevant anodic peak current varies linearly with cysteine concentration within the range 9.0x10(-4) to 1.0x10(-2) mol L(-1).

Calibration↗

PubMed2004

Study of the lipophilicity of selenium species.

In this work the lipophilicity of different selenium species occurring in environmental matrices and food, Se(IV), Se(VI), selenomethionine (Se-Met), selenocystamine (Se-CM), selenocystine (Se-Cyst), and dimethyl diselenide (CH3)2Se2, was investigated in the octanol-water system, using the shaking flask method and detection with inductively coupled plasma-atomic emission spectrometry (ICP-AES), in order to assess their environmental fate and tendency to bioaccumulate. Polarography was also used for the electrochemically active Se species, Se(IV), Se-Cyst, Se-CM and (CH3)2Se2, and the results were compared with those measured by ICP-AES. Furthermore, the influence of pH was studied by determining the logarithm of the distribution coefficient, log D, at three pH values, 5, 7, and 9, as was the impact of the marine environment on the lipophilicity profile of the six investigated Se species. The results were compared with those estimated approximately by use of PrologD software, based on the Ghose-Crippen log P (P: partition coefficient) calculation system, the only system which incorporates values-even though approximate-for the atom type of Se. Finally, from our experimental data an indicative value of the Se-Se fragment for log P prediction, for use in drug design, was estimated.

Hydrogen-Ion Concentration↗

PubMedDate not supplied

Copper-complexation index as a polarographic summation parameter for the determination of chelating agents in water.

Polarography of the copper(II)-complexes of chelating agents is shown to be a suitable basis for a summation parameter for the total concentration of these chemicals in water (expressed as the copper-complexation index). Using 12 chelating agents (ATMP, BAED, CDTA, DTPA, DTPMP, EDTA, EDTMP, EDTP, EGTA, HEDP, NTA, PBTC) all listed in DIN 38409-H26 and DIN 38413-P5 a satisfactory reproducibility of the method used was obtained. In addition several commercial tensides, detergents in washing and deliming products were investigated.

Journal Article↗

PubMed2001

Electrochemical monitoring of the thermal decomposition of arenediazonium ions with time in acidic MeOH/H2O mixtures: dediazotization rate constant.

The rate of the spontaneous decomposition of 4-methylbenzenediazonium tetrafluoroborate, 4MBD, was determined. The effects of solvent composition (MeOH/H2O) on its electrochemical processes and on those of the derivatized 2-naphthol-6-sulfonate-4-toluene (6S2NPT) azo dye were investigated by Differential Pulse Polarography (DPP). This electrochemical technique shows an effective sensitivity for detecting arenediazonium ions and derivatized azo dyes.

Journal Article↗

PubMed2001

Electrochemical methods for monitoring of environmental carcinogens.

The use of modern electroanalytical techniques, namely differential pulse polarography, differential pulse voltammetry on hanging mercury drop electrode or carbon paste electrode, adsorptive stripping voltammetry and high performance liquid chromatography with electrochemical detection for the determination of trace amounts of carcinogenic N-nitroso compounds, azo compounds, heterocyclic compounds, nitrated polycyclic aromatic hydrocarbons and aromatic and heterocyclic amines is discussed. Scope and limitations of these methods are described and some practical applications based on their combination with liquid-liquid or solid phase extraction are given.

Azo Compounds↗

PubMed1996

Complexation capacity of aquatic systems in dependence on different ligands and heavy metals - Electroanalytical investigations and statistical evaluation.

Heavy metals exist in natural waters in different species. Mobility and bioavailability of bound metals can be influenced by some complex factors in the aquatic environment. Therefore, it is useful to determine the complexation properties of various ligands influenced by different metals. The copper and zinc complexation capacity was determined for a natural ligand (fulvic acid) and anthropogenic pollutants (e.g. nitrilotriacetic acid and ethylenediaminetetraacetic acid). The interactions between several ligands and metal ions and their effects on the complexation capacity were analyzed in particular. Applying methods of experimental design, such as multifactorial plans, it is possible to determine influence and interaction of various parameters (e.g. concentration of zinc or copper) by a minimum number of experiments. Differential pulse polarography was used for the determination of the complexation capacity. Methods of parametric and robust multiple linear regressions were applied for the interpretation of the measured values.

Journal Article↗

PubMed1996

Polarographic and voltammetric investigations of new catalytic systems of iron for application in trace analysis.

New catalytic systems consisting of complexes of Fe(III)-ions with ligands such as nitrilotriacetic acid, N-(2-hydroxyethyl)iminodiacetic acid, N,N-bis(2-hydroxyethyl)glycine, triisopropanolamine, 2-hydroxy-1,3-diaminopropanetetraacetic acid, tris-(hydroxymethyl)aminomethane, bis-(2-hydroxymethyl) iminotris-(hydroxymethyl)methane, 1,3-bis/tris(hydroxymethyl)methylamino/propane in ammonia buffer and 1,2-dihydroxybenzene in tris-(hydroxymethyl) amino-methane-HCl buffer and ClO(-)(2)-ions as the oxidizing agent were studied by pulse polarography and voltammetry. Optimal conditions such as kind and concentration of the ligand, the composition of supporting electrolyte and concentration of NaClO(2) for the determination of traces of iron in the presence of a large excess of other electrochemically active ions were examined. Examples of acceleration and inhibition of catalytic reactions between Fe(II). L and ClO(-)(2)-ions by electrochemically inactive ions have been found.

Journal Article↗

PubMed2002

Severe depletion of mitochondrial DNA in spinal muscular atrophy.

Spinal muscular atrophy (SMA) is a neuromuscular disorder in childhood leading to a dramatic loss of muscle strength. Functional investigations with high-resolution polarography and enzyme measurements of the respiratory chain revealed lowered activities in muscle tissue of SMA patients. To gain a better understanding of this low energy supply we analyzed the amount of mitochondrial DNA (mtDNA) in skeletal muscle of 20 unrelated children with genetically proven SMA and 31 controls. Quantitative Southern blot analysis revealed a severe and homogeneous decrease in the content of muscle mtDNA in relation to nuclear DNA in SMA patients (90.3+/-7.8%), whereas by immunofluorescence no decrease in the number of mitochondria was detected. In addition, a two- to threefold reduction of the nuclear-encoded complex II (succinate dehydrogenase) activity was detected in SMA muscle tissue. Western blot analysis showed a significant reduction of both mitochondrial- and nuclear-encoded cytochrome c oxidase subunits. Our results indicate that mtDNA depletion in SMA is a consequence of severe atrophy, and has to be differentiated by measurement of complex II from an isolated reduction of mtDNA as found in patients with mitochondriocytopathies and the so-called mtDNA depletion syndrome.

Adolescent↗

PubMed2003

In vitro percutaneous absorption of cobalt.

OBJECTIVES: To evaluate skin absorption of cobalt powder in an in vitro system. Experiments with volunteers show that cobalt powder can permeate through the skin, but there are no data with regard to the mechanism or the amount of permeation. METHODS: Skin permeation was calculated by the Franz diffusion cell method with human skin. A physiological solution was used as receiving phase and the cobalt powder was dispersed in synthetic sweat. The amount of metal passing through the skin was analysed by electro-thermal atomic absorption spectrometry (ETAAS). Parallel polarographic analysis (differential pulse polarography-DPP) allowed evaluation of cobalt present as ions (Co(2+)) in donor and receiving phases. Measurements of cobalt skin content were also performed. RESULTS: Evaluation of metal in the receiving phase allowed us to demonstrate the permeation of cobalt through the skin. Steady-state flow of percutaneous cobalt permeation was calculated as 0.0123+/-0.0054 microg cm(-2) h(-1), with a lag time of 1.55+/-0.71 h. CONCLUSIONS: The experiments show that cobalt powder can pass through the skin when applied as a dispersion in synthetic sweat, oxidising metallic cobalt into ions, which permeate the skin. These experiments show for the first time how cobalt can permeate the skin.

Cobalt↗

PubMed1999

Tumor necrosis factor-alpha plus actinomycin D-induced apoptosis of L929 cells is prevented by nitric oxide.

Treatment with the nitric oxide-(NO)-generating compound S-nitroso-N-acetylpenicillamine protected cul-tured L929 cells from apoptosis induced by tumor necrosis factor-alpha (TNF-alpha) plus actinomycin D, as determined by the detection of DNA fragmentation and morphological changes. NO also prevented an enhancement of the production of reactive oxygen intermediates by TNF-alpha plus actinomycin D, as assessed by the oxidation of dihydrorhodamine 123 and hydroethidine. Because the inhibition of mitochondrial respiration by rotenone or antimycin A suppressed the increased oxidation of both dihydrorhodamine 123 and hydroethidine, it was suggested that TNF-alpha accelerated the leakage of reactive oxygen intermediates from the mitochondrial electron transport system. Polarography showed that NO reversibly inhibited mitochondrial respiration at either complexes I-III, II-III, or IV, thus suggesting the inhibition of cytochrome oxidase. Taken together, these findings indicate that the decreased mitochondrial formation of reactive oxygen intermediates in the presence of NO might have a protective effect against TNF-alpha plus actinomycin D-induced apoptosis.

Animals↗

PubMed2002

Fasting induces cyanide-resistant respiration and oxidative stress in the amoeba Chaos carolinensis: implications for the cubic structural transition in mitochondrial membranes.

Large free-living amoeba (Chaos carolinensis) can survive in spring water without food intake for several weeks. Starvation is associated with a dramatic change in mitochondrial cristae from random tubular to ordered (paracrystalline) cubic morphology. Whole-cell polarography was used to monitor changes in respiratory activity during fasting. Basal respiration per cell decreased progressively during starvation, while the cyanide-resistant fraction increased. Spectrofluorometric assay of H2O2 and reactive oxygen species (ROS) in cell lysates (using the dye 2',7'-dichlorofluorescein diacetate) indicates greater H2O2 and ROS generation in starved than in fed cells. Fluorescence microscopy of intact cells incubated with the same dye demonstrates that H2O2 and ROS tend to accumulate in vacuoles. A remarkable generation of O2 observed with starved cells after addition of KCN may be explained by release of H2O2 from these compartments into the cytosol, where it can react with catalase. Together, these observations suggest that fasting increases oxidative stress in the amoeba and that this organism has several protective mechanisms to deal with it, including activation of a plantlike alternative oxidase. The hypothesis is forwarded that the cubic structural transition of the mitochondrial inner membrane represents another protective mechanism, reducing oxidative damage by enhancing the efflux of H2O2 and ROS and by reducing the susceptibility of membrane lipids to the oxidants.

Amoebida↗