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[Determination of Ag ions in water, urine and blood].

Developments in polarography techniques over the last thirty years have enabled us to register outstanding increases in sensitivity: the order of concentration studies has evolved from 10(-3) g ions/1 to 10(-10) g ions/1. Pulse polarography with anodic stripping voltammetry is therefore particulary suitable for the determination of numerous metals which are to be found in extremely low concentration in biological samples which may be of limited volume e.g. blood. Our objects has been to describe and to apply in the case of silver a determination technique which would be sufficently sensitive, precise, practical and rapid for it to be easily used by biologists and toxicologists. A fortiori, this method may be applied in hydrology to the determination of these metals in water.

Humans↗

Hair sulfur amino acid analysis.

This overview emphasizes present aspects of sulfur-containing amino acids in hair. A selection of analytical procedures to determine cystine, cysteine, S-sulfocysteine, cystine oxide, cysteic acid, lanthionine and lysinoalanine are presented. The methods relate to intact hair or partial and total hydrolysates and comprise chromatography, titration, colorimetry, polarography and spectroscopy. For the analysis of cysteine, cystine and cystine oxides, polarography and spectroscopy are the methods of choice. Cysteic acid, lanthionine and lysinoalanine are analysed by means of ion-exchange chromatography (Spackman et al., 1958) after total hydrolysis.

Alkalies↗

Effects of evening primrose oil treatment on sciatic nerve blood flow and endoneurial oxygen tension in streptozotocin-diabetic rats.

Evening primrose oil (EPO) is rich in the omega-6 essential fatty acid component, gamma-linolenic acid. The aim of the investigation was to determine whether EPO treatment prevented a reduction in sciatic nerve perfusion and oxygenation in streptozotocin-diabetic rats. Rats were treated from diabetes induction with 10 g EPO kg-1 day-1. Sciatic blood flow was measured by microelectrode polarography and hydrogen clearance. Diabetes caused 47.7% +/- 3.4% (P < 0.001) and 58.8% +/- 4.8% (P < 0.001) reduction in the nutritive (capillary) and the non-nutritive (large vessel) components of endoneurial blood flow, respectively, which were prevented by EPO. Treatment had no significant effect on nutritive flow in non-diabetic rats; however, the rate of non-nutritive flow increased by 97.7% +/- 38.9% (P < 0.01). Sciatic endoneurial oxygen tension was measured by microelectrode polarography. Diabetes resulted in a 44.7% +/- 3.4% reduction in mean oxygen tension (P < 0.001), which was largely (82.3% +/- 10.2%) prevented by EPO treatment (P < 0.001). Thus, EPO prevents impairment of blood flow and endoneurial oxygenation in experimental diabetes. It is likely that this neurovascular action accounts for the beneficial effects of treatment on nerve function in diabetic rats and patients.

Animals↗

Polarographic and voltammetric determination of trace amounts of 2-nitronaphthalene.

The polarographic behaviour of 2-nitronaphthalene was investigated by DC tast polarography (DCTP) and differential pulse polarography (DPP), both at a dropping mercury electrode, and differential pulse voltammetry and adsorptive stripping voltammetry, both at a hanging mercury drop electrode. Optimum conditions have been found for the determination of 2-nitronaphthalene by the given methods in the concentration ranges of 2 x 10(-6) - 1 x 10(-4), 2 x 10(-7) - 1 x 10(-4), 1 x 10(-8) - 1 x 10(-4) and 2 x 10(-9) - 1 x 10(-8) M, respectively. Practical applicability of these techniques was demonstrated by the determination of 2-nitronaphthalene in drinking and river water after its preliminary separation and preconcentration using liquid-liquid and solid-phase extraction with limits of determination of 3 x 10(-10) M (drinking water) and 3 x 10(-9) M (river water).

Electrochemistry↗

Evidence for intramitochondrial complementation between deleted and normal mitochondrial DNA in some patients with mitochondrial myopathy.

Twenty-three patients with mitochondrial myopathies and mitochondrial DNA deletions in muscle were studied by means of deletion mapping and sequencing, histochemistry and polarography. Histochemistry showed significantly less focal cytochrome oxidase deficiency relative to number of ragged red fibres when the deletion did not involve reading frames for cytochrome oxidase subunits. Polarography in such patients showed defects exclusively involving complex I, in contrast to the others with larger deletions who generally had more diffuse respiratory chain defects. Analysis of other published histochemical data showed similar findings to our own. It is concluded that translation of a proportion of deleted mitochondrial DNAs occurs in at least some patients with mitochondrial DNA deletions, implying that deleted and normal mitochondrial genomes share transfer RNAs within mitochondria in such cases.

Base Sequence↗

Interaction of nucleic acids with electrically charged surfaces. VII. The effect of ionic strength of neutral medium on the conformation of dna adsorbed on the mercury electrode.

Triangular-wave direct current (d.c.) voltammetry at a hanging mercury drop electrode and phase-selective alternating current (a.c.) polarography at a dropping mercury electrode were used for the investigation of adsorption of double-helical (ds) DNA at mercury electrode surfaces from neutral solutions of 0.05-0.4 M HCOONH4. It was found for the potential region T (from -0.1 V up to ca. -1.0 V) that the height of voltammetric peaks of ds DNA is markedly influenced by the initial potential only at relatively low ionic strength (mu) (from 0.05 up to ca. 0.3). Also a decrease of differential capacity (measured by means of a.c. polarography) in the region T depended markedly on the electrode potential only at relatively low ionic strength. The following conclusions were made concerning the interaction of ds DNA with a mercury electrode charged to potentials of the region T in neutral medium of relatively low ionic strength mu < 0.3). (i) When ds DNA is adsorbed, a significantly higher number of DNA segments is anchored in the positively charged electrode surface than in the surface bearing a negative charge, (ii) In the region T, especially adsorbed labile regions of ds DNA are opened in the electrode surface, which are present in ds DNA already in the bulk of the solution, (iii) In the narrow region of potentials in the Vicinity of the zero charge potential a higher number of ds DNA segments can be opened, probably as a consequence of the strain which could act on the ds DNA molecule in the course of the segmental adsorption/desorption process.

Journal Article↗

Voltammetric study of danazol and its determination in capsules and spiked biological fluids.

The voltammetric behaviour of danazol DZ (antigonadotropin) was studied using cyclic voltammetry, direct current, differential pulse polarography (DPP) and alternating current polarography. Danazol exhibited irreversible cathodic waves over the pH range of 1-5 in Britton Robinson buffers. At pH 1 (the analytical pH), a well-defined wave with E1/2 of -1.04 V versus Ag/AgCl reference electrode was obtained. The diffusion current constant (Id) was 4.8+/-0.14 microA.L.m mole(-1) and the current-concentration plot was rectilinear over the range from 5 x 10(-6) to 1 x 10(-4) M with correlation coefficient (n = 11) of 0.995. The calculated detection limit was 1 x 10(-6) M using the DPP mode. The wave was characterized as being irreversible, diffusion-controlled although adsorption phenomenon played a limited role in the electrode process. The proposed method was applied to commercial capsules and the average percentage recovery was in agreement with that obtained by the official USP method. The method was extended to the in vitro determination of DZ in spiked human urine and plasma samples, the percentage recoveries were 96+/-4 and 97+/-5, respectively. A proposal of the electrode reaction was postulated.

Adult↗

Electrochemical behaviour and determination of acrivastine in pharmaceuticals and human urine.

The differential pulse polarography, DC-tast polarography and cyclic voltammetry behaviour of acrivastine was studied in Britton-Robinson buffer solutions (pH 2-11.7). In acidic media, a non-reversible diffusion controlled reduction process involving four electrons takes place. Two reduction waves appear at a E(1/2)=-0.6 and -0.99 V. The reduction mechanism is discussed. The linear relationship between peak current height and acrivastine concentration allowed the differential pulse polarographic determination of acrivastine over a wide concentration range, from 0.35 to 26.1 mg l(-1)at pH 2.5. The procedure was applied to determination of the drug in pharmaceutical formulations and human urine samples.

Electrochemistry↗

Electrochemical and spectroelectrochemical behavior of the main photodegradation product of nifedipine: the nitrosopyridine derivative.

PURPOSE: To characterize the electrochemical behavior of the photodegradation product of nifedipine, i.e., 2,6-dimethyl-4-(2-nitrosophenyl)-3,5-pyridine-carboxylic acid dimethyl ester (NPD) in different electrolytic media. We also evaluated the interaction between free radicals generated from NPD and xeno/endobiotics. METHODS: Tast polarography, differential pulse polarography, and cyclic voltammetry were used for the characterization. Controlled potential electrolysis and ultraviolet-visible spectroscopy were used to generate and to detect the nitroso radical anion. RESULTS: In protic media, the NPD derivative gave a reversible well-defined peak either on Hg or glassy carbon electrodes in a reaction involving two electrons and two protons to give the hydroxylamine derivative. In mixed aqueous-organic media (pH 9) and in aprotic media, nitroso radical anion was isolated and characterized, exhibiting second-order dimerization rate constant (k2) values of 11,300 +/- 210 [Ms](-1) and 8,820 +/- 78 [Ms](-1), respectively. Reactivity of the nitroso radical anion with relevant pharmacologic targets revealed a significant interaction with the tested endo/xenobiotics (cysteamine, GSH, N-acetylcysteine, and adenine). CONCLUSIONS: Both in mixed and aprotic media, NPD generated free-radical species, the nitroso radical anion. Taking into account their respective interaction rate constants, the following tentative rank order of reactivity can be established as follows: cysteamine > N-acetylcysteine > GSH > adenine.

Electrochemistry↗

Electrochemical characteristics of five quinolone drugs and their effect on DNA damage and repair in Escherichia coli.

The object of this study was to determine whether 4-quinolone antimicrobials were reduced under biologically attainable redox conditions and whether they had any effect on DNA in the absence of the DNA gyrase enzyme. Electrochemical characteristics of the drugs were investigated using d c polarography, differential pulse polarography and cyclic voltammetry. The ability of the drugs to interact with, and cause damage to, naked DNA was investigated by a phi X174 DNA double transfection assay. Induction of DNA SOS repair was assessed using a stain of Escherichia coli in which the synthesis of beta-galactosidase was under the control of the su1A gene. Growth studies were performed using a conductimetric method in a Malthus system. All five 4-quinolones examined had redox potentials lower (more negative) than -1.2 V and thus were incapable of being reduced in biological systems, even under strict anaerobiosis. Exposure of all drugs to single-stranded phi X174 DNA for up to 50 h engendered no detectable damage. However, all the drugs induced DNA SOS repair, in the order ciprofloxacin greater than fleroxacin = pefloxacin greater than norfloxacin greater than nalidixic acid. This rank order corresponds approximately with antibacterial efficiency. The growth studies indicated that redoxyendonuclease III and excision repair enzymes may be involved in the fixation of quinolone-induced damage.

4-Quinolones↗

Electrochemical behavior and detection of plant hormone 6-benzyl adenine in acetate buffer at mercury electrode.

The electrochemical behavior of 6-benzyl adenine (6-BA) has been studied in 0.1 mol L(-1) HAc-NaAc solution (pH 3.8). Cyclic voltammetry, single-sweep polarography and direct current polarography were employed to clarify the mechanism of the electrode process; the kinetic parameters of the rate-determining step were determined. Reduction of 6-BA involves two pH-dependent processes, corresponding to the overall irreversible reduction of the 1,6 and 3,2 N=C bonds. Each reduction stage consists a preprotonation of the nitrogen atom at the electroactive site and a rapid two-electron transfer. In the presence of 6-BA, the reduction potentials of some ions were shifted. Under the given conditions, 6-BA displays one irreversible reduction peak controlled by adsorption. Two linear response were observed in the range 2.0 x 10(-8) - 8.0 x 10(-7) mol L(-1) and the range 1.0 x 10(-6) - 8.0 x 10(-6) mol L(-1), with correlation coefficients of 0.9995 and 0.9998, respectively. The detection limit is 7.1 x 10(-9) mol L(-1). The method had been applied to the determination of 6-BA in bean sprout samples with satisfactory results.

Acetic Acid↗

[Characteristics of the blood supply of suppurative wounds treated by a defocused beam of a CO2 laser].

In examination of 57 patients with the use of a method of complex polarography, by oxygen and hydrogen, the increase in volumetrical rate of blood flow and oxygen tension in the purulent wounds treated by a defocused beam of the CO2 laser was established. This contributed to more favourable course of the wound process and accelerated healing. Complex polarography can be used as a diagnostic method in the purulent surgery.

Adolescent↗

[The characteristics of the trophic neuromicrocirculatory interrelationships in traumatic neuropathies of the upper extremities].

Neurotrophic effects on tissues microcirculation in upper limbs traumas were revealed by 9-year investigations using electroneuromyography, capillaroscopy of the nail matrix, polarography, oxygen test, hydrogen clearance test. It is shown that oxygen polarography is most adequate for evaluation of nerve trophism. A total of 136 patients with neuropathies of 174 nerves have been examined. Of principal importance for the results interpretation is examination of the neurosecretory function of thin nonmyelinated fibers and vasoactive neuropeptides in neuromicrocirculatory interactions. The action potential of neurotrophic substances depends on their tissue levels.

Arm↗

Electrostatic and hydrophobic interactions are involved in factor Va binding to membranes containing acidic phospholipids.

The interaction of factor Va with phospholipid monolayers was studied using alternating current polarography. For these studies a hanging mercury drop electrode is positioned in contact with the monolayer at the air-monolayer interface. Factor Va introduced into the aqueous subphase beneath the monolayer caused alterations in the capacitance of the circuit. When factor Va was introduced beneath monolayers containing 13-25% mole fraction phosphatidylserine (PS), 87-75% mole fraction phosphatidylcholine (PC), a significant capacitance increase occurred, which is interpreted to be the result of the penetration of the factor Va molecules through the monolayer. No penetration or associated capacitance changes were observed with monolayers composed of pure PC or pure PS. Polarography experiments were also conducted with the electrode placed in the aqueous subphase to measure the binding of factor Va to pure PS monolayers. This approach has allowed detection of factor Va (at nM concentrations) adsorption to a pure PS monolayer. Thus the membrane adsorption process can be distinguished from factor Va penetration through the monolayer. These results suggest that there are two types of interaction of the factor Va molecule with PS-containing monolayers. The interaction with the PS component of the monolayer is essential for binding the factor Va (at nM concentrations) that occurs by surface adsorption. For mixed PC/PS monolayers, PS adsorption is followed by penetration of the protein through the membrane.

Animals↗

[Drug analysis with polarographic methods. 33. Electroanalytic studies of a new benzodiazepine: [6(o-chlorophenyl)-2,4-dihydro-2-[(4-methyl-1-piperazinyl) methylene]-8-nitro-1H-imidazol[1,2-a][1,4]benzodiazepine-1-one (WHO) (loprazolam,1)] and content determination its drug forms].

Loprazolam (1) is a tricyclic benzodiazepine containing a new butazadiene moiety, which has not been investigated by polarography up to now. 1 is reduced in three waves at a DME over the whole pH-region. In BRP (pH 2-9) 10 electrons are consumed in this process. The first step is suitable for the determination of 1 in Loprazolam tablets containing 1 or 2 mg. These tablets are on the pharmaceutical market in several European countries. The mechanism of the electrode process will be reported in the communication XXXIV.

Anti-Anxiety Agents↗

Extraction and analysis of superoxide free radicals (.O2-) from whole mammalian liver.

Extraction of whole lobes of normal rat liver with dimethyl sulphoxide (DMSO) under N2 gives extracts which contain 5-10 mumol/l.O2- (50-100 nmol.O2- per 10 ml extract per 4 g liver; 1.25-2.50 nmol.O2- per millilitre per gram liver). Evidence for .O2- in the extracts is given by: (1) electron spin resonance signals (ESR), (2) differential pulse polarography (DPP), (3) chemiluminescence (CL), and (4) nitroblue tetrazolium reduction (NBT). All tests yield results identical with those obtained with authentic .O2-. Extraction of .O2- is enhanced by tetrabutyl ammonium ion, and is maximal at 1-3 min. These results raise the possibility that substantial amounts of .O2- are normally sequestered in protective membranous sites in vivo.

Animals↗

An improved method for isolation and identification of Zn-metallothionein from cadmium-induced rat liver.

Zn-metallothioneins (MT-1 and MT-2) were isolated and purified from Wistar rat liver induced by subcutaneous injection with cadmium chloride over a short time. Instead of Sephadex G-50 and DEAE Sephadex A-50, new chromatographic media produced by Pharmacia, Sephacryl S-200, S-100 and DEAE Sepharose Fast Flow were used in the purification of metallothioneins. The time required for purification with the new method was only 1/3 that required with the usual method and had the same purification effect and rate of recovery. The number of mercapto groups measured with modified Ellman's reagent and cysteine as standard is 20 in MT molecules. Zn and Cd concentrations in each fraction were measured by single sweep polarography rather than atomic absorption spectrophotometry. MT-1 and MT-2 contained 6 gram atoms of zinc, but no cadmium. Purified MT-1 and MT-2 were shown by high performance liquid chromatographic analysis to be highly homogeneous and had an amino acid composition similar to that of Cd-MT.

Amino Acids↗

[Square-wave polarographic determination of heavy metals in vegetables with special regard to cadmium].

The cadmium contents of 104 foods mostly of vegetable origin (market samples from a highly industrialized area in the GDR) were determined by means of square-wave polarography. The values obtained are in good agreement with literature data about the cadmium contents of foods, and they are not indicative of an increased contamination of the products under investigation compared to other industrialized countries.

Cadmium↗