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At least 343 records · Page 19Linked to original sources
PubMed1986

A new membrane-bound b-type cytochrome, cytochrome b-558, from photosynthetically grown Rhodopseudomonas sphaeroides.

A new membrane-bound b-type cytochrome, cytochrome b-558, was removed from chromatophore membranes of photosynthetically grown Rhodopseudomonas sphaeroides strain R-26 by deoxycholate-cholate extraction. The cytochrome was purified by ammonium sulfate fractionation and ion-exchange chromatography. Cytochrome b-558 had absorption maxima at 280 and 405 nm in the oxidized form, and at 558, 528, and 420 nm in the reduced form. It had a midpoint potential of--130 mV at pH 7.0. The minimal molecular weight of this protein was 42,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and it contained one mole heme per mole of protein. The isoelectric point was 8.5. The electrophoretic pattern of heme-carrying proteins and the redox potentiometry showed that cytochrome b-558 was present in membranes from wild type, strain R-26, and strain GA grown photosynthetically, but not from any strain grown aerobically.

Amino Acids↗
PubMed1976

Iron-sulfur proteins of the green photosynthetic bacterium Chlorobium.

The iron-sulfur proteins of the green photosynthetic bacterium Chlorobium have been characterized by oxidation-reduction potentiometry in conjunction with low-temperature electron paramagnetic resonance spectroscopy. Chlorobium ferredoxin was the only iron-sulfur protein detected in the soluble fraction; no high-potential iron-sulfur protein was observed. In addition, high-potential iron-sulfur protein was not detected in the chromatophores. Four chromatophore-bound iron-sulfur proteins were detected. One is the "Rieske" type iron-sulfur protein with a g-value of 1.90 in the reduced state; the protein has a midpoint potential of + 160 mV (pH 7.0), and this potential is pH dependent. Three g=1.94 chromatophore-bound iron-sulfur proteins were observed, with midpoint potentials of -25, -175, and about -550 mV. A possible role for the latter iron-sulfur protein in the primary photochemical reaction in Chlorobium is considered.

Binding Sites↗
PubMed1980

An electrochemical study of the iron storage protein, ferritin.

The electrochemical behavior of iron storage protein, ferritin (molecular weight 700 000) was studied by pulse polarography, cyclic voltammetry and 'potentiometry' monitored by colorimetry. Comparison between in vivo and voltammetric redox reactions of the ferritin Fe(III) core and thermodynamic E'0 values for native ferritin are described.

Electrochemistry↗
PubMed1994

The structure and (local) stability constants of borate esters of mono- and di-saccharides as studied by 11B and 13C NMR spectroscopy.

The formation of borate esters of various mono- and di-saccharides in aqueous solution was studied by 11B and 13C NMR spectroscopy. Association constants K(B-L) at a carbohydrate-borate molar ratio of 1:1, pH 7, and 25 degrees C were determined and compared with literature values obtained from potentiometry. The association constants K(B-L) were converted into local association constants Kloc(B-L) by using the distribution of the various anomeric forms in D2O. In this way, values of Kloc(B-L) were obtained, which appear to be characteristic of the configuration concerned. They explain the favourable effect of borate in the alkaline isomerisation of lactose into lactulose. At a low molar ratio (1:3) of carbohydrate-borate, predominantly diborate esters (B-)2L were formed.

Borates↗
PubMed1988

Measurement of ionized calcium in biological fluids: ion-selective electrode method.

There is now a consensus that Ca2+ measurements are more physiologically and clinically meaningful than CaT measurements. Ca2+ in serum, plasma, whole blood, and other biological fluids can be measured by direct potentiometry with ion-selective electrodes and a number of reliable instruments are commercially available for this measurement. Several factors affect the Ca2+ concentration and must be carefully controlled for the results to be meaningful. The most important of these considerations are the anaerobicity of the sample, the need to concurrently measure pH, and the concentration of heparin, if whole blood or plasma samples are used. The calibration of the Ca2+ ISE is critical to the accuracy of the measurement. The matrix of the calibrator should match that of the sample as closely as possible, particularly in regard to ionic strength and liquid junction potential. The measurement of Ca2+ in urine is complicated by the wide variation in ionic strength encountered in this type of sample; thus, it is more meaningful to standardize the ISE in terms of Ca2+ activity instead of concentration. Instrumentally, the measure of copper in biological samples can be achieved with high accuracy, high precision, without background correction, and with minimum sample pretreatment if care is taken to carefully plan and implement all the critical steps in the analysis procedure.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1993

Relation between the physicochemical characteristics of collagen and its interactions with chitosan: I.

The interaction between bovine atelocollagen and a high molecular weight fully deacetylated chitosan has been studied. Considering the apparent mass of an anionic equivalent of collagen and the results of various techniques (potentiometry, conductometry and IR spectroscopy), a purely electrostatic mechanism is demonstrated. The role of charge density and chain length of collagen chains is discussed, and although the interaction is weak and hindered by gel formation in collagen solutions, a polyanion-polycation complex is obtained. Circular dichroism spectroscopy experiments show an organization in the solid state of the complex which is quite different to those of collagen and chitosan.

Acetylation↗
PubMed1984

Trace metal requirements in total parenteral nutrition (TPN). 5. Formation constants for the copper(II)--histidine ternary complexes with threonine, lysine, glycine, phenylalanine, valine, and cystine, and discussion of their implications regarding the copper distribution in blood plasma during TPN and the evaluation of the daily dose of copper.

Specific metal deficiencies have been reported to affect patients receiving total parenteral nutrition (TPN). Our previous studies on the topic were devoted to the computer-based interpretation of the extra urinary excretion of zinc; a theoretical approach was also proposed, with a view to compensating for the extra losses of this metal. Similarly, the present work deals with the problem of TPN-induced copper deficiency and its remedy. As is the case for zinc, the TPN-induced excretion of copper clearly stems from the relative mobilization of the plasma protein-bound pool of this metal into its diffusable low-molecular-weight fraction; this phenomenon being due to the competitive complexation of copper by the amino acids of the nutritive solution. The computer simulation of this effect thus required that first the equilibrium constants be experimentally determined for the main complexes of copper that might form in the solution as well as in plasma during the infusion. Accordingly, complex formation in the copper-histidine ternary systems with threonine, lysine, glycine, phenylalanine, valine, and cystine was investigated by potentiometry at 37 degrees C in NaCIO4 0.15 mol X dm-3. The implications of the results obtained are discussed with regard to the interpretation of the copper excretion and the estimation of the desirable daily dose of this metal for the TPN mixture under consideration.

Amino Acids↗
PubMed1989

Phytic acid-enhanced metal ion exchange reactions: the effect on carboxypeptidase A.

On the basis of the known interaction of phytic acid to form soluble or insoluble complexes with cations, the effect of this naturally occurring polydentate ligand on carboxypeptidase A, a zinc-containing metalloenzyme, and its Co(II)-substituted derivative, has been studied. Under conditions of rigorous exclusion of adventitious metal ions, phytate showed no inhibitory effect. However, the addition of Cu(II) ions to form soluble phytate-Cu(II) complexes at pH 7.2 and 25 degrees C caused more than a 95% decrease in activity. The Cd(II) ion was nearly as effective but other ions showed only a small or no effect. In the absence of phytate, incubation of the enzyme with Cu(II) or Cd(II) at the same concentration produced only about a 25% reduction in activity. The decrease in activity followed first-order kinetics, and the rate constant was the same (1.2 x 10(-4) sec-1) as seen upon incubation with EDTA. However, in contrast to that observed upon incubation of the enzyme with phytate and Cu(II), exposure to EDTA produced a complete loss in activity which could be regained by addition of Zn(II) to the assay solution. In the former case, not only was there residual activity left after incubation at pH 7.2 for 24 hrs at 25 degrees C, but the initial activity could not be regained under similar assay treatment. An increase in either the Cu(II) or phytate concentration while the other was kept constant, yielded saturation curves with maximal effect at 3 x 10(-5) M for Cu(II) and at 5 x 10(-5) M for phytate (enzyme at ca. 10(-6) M). At these ratios, all of the cupric ions are completely bound to phytate as determined by ion-selective potentiometry. A preparative scale reaction of phytate and Cu(II) with carboxypeptidase A (kcat 8460 min-1; K'M 0.23 mM with CBZ-glycyl-glycyl-L-phenylalanine as substrate at pH 7.5, 25 degrees C) gave a product isolated in 95% yield but with lower activity (kcat 198 min-1; K'M 0.25 mM). A Cu(II)-carboxypeptidase preparation had similar kinetic parameters (kcat 207 min-1; K'M 0.34 mM). This near identity of constants suggested that a metal exchange reaction had occurred, i.e., incubation of Zn(II)-carboxypeptidase with a phytate-Cu(II) complex resulted in not only the removal of the zinc ion from the active site but also the sequential and rapid incorporation of a cupric ion into the apoenzyme so formed.(ABSTRACT TRUNCATED AT 400 WORDS)

Cadmium↗
PubMed1990

Influence of respiratory substrate on the cytochrome content of Shewanella putrefaciens.

Shewanella putrefaciens can use trimethylamine oxide (TMAO) as electron acceptor under anoxic conditions. The associated cytochromes induced during growth under various respiratory conditions have been separated by liquid chromatography (DEAE Sepharose CL6b) and SDS-PAGE and characterized spectrophotometrically and by redox potentiometry. Two major low potential cytochromes and at least three minor low potential cytochromes, likely to be involved in TMAO reduction, were found. No cytochrome specific for TMAO reductase was found.

Chromatography, High Pressure Liquid↗
PubMed1983

Recent applications of electrochemical techniques to the analysis of pharmaceuticals.

The past decade has revealed a remarkable and renewed interest in the application of electrochemical techniques to the analysis of drugs and pharmaceuticals. In addition, several of these techniques are now being employed to elucidate electrode processes by studying the mechanisms involved in organic redox reactions. The renewed interest in electrochemical techniques can be attributed in part to more sophisticated instrumentation and to an increased understanding of the techniques themselves. This paper selectively reviews recent applications of the following electrochemical modes in the analysis of drugs and pharmaceuticals: amperometry, conductometry, coulometry, ion-selective electrodes, potentiometry, stripping voltammetry (anodic and cathodic) and voltammetry.

Journal Article↗
PubMed1985

A quinidine-responsive plastic membrane electrode.

A membrane electrode based on quinidine tetraphenylborate in a PVC matrix is described. The electrode exhibits a rapid and near-Nernstian response in the range 3.5 x 10(-5)-1 x 10(-2) M quinidine sulphate at pH 6-8. In an acidic medium the electrode responds to diprotonated quinidine. In sodium tetraphenylboron (Na TPB) solutions the response is linear in respect of log (TPB-) over the range 10(-4)-10(-2)M . Direct potentiometry and potentiometric titrations are used to determine quinidine in pharmaceutical preparations.

Journal Article↗
PubMed2005

Direct electron transfer reactions of laccases from different origins on carbon electrodes.

Electrochemical studies of laccases from basidiomycetes, i.e., Trametes hirsuta, Trametes ochracea, Coriolopsis fulvocinerea, Cerrena maxima, and Cerrena unicolor, have been performed. Direct (mediatorless) electrochemistry of laccases on graphite electrodes has been investigated with cyclic voltammetry, square wave voltammetry as well as potentiometry. For all mentioned high potential laccases direct electron transfer (DET) has been registered at spectrographic graphite and highly ordered pyrolytic graphite electrodes. The characteristics of DET reactions of the enzymes were analysed under aerobic and anaerobic conditions. It is shown that the T1 site of the laccase is the primary electron acceptor, both in solution (homogenous case) and at surface of the graphite electrode (heterogeneous case). A mechanism of ET for the process of the electro-reduction of oxygen at the laccase-modified graphite electrodes is proposed and the similarity of this heterogeneous process to the laccase catalysed oxygen reduction homogeneous reaction is concluded.

Basidiomycota↗
PubMed2006

Preliminary electrochemical studies of the flavohaemoprotein from Ralstonia eutropha entrapped in a film of methyl cellulose: Activation of the reduction of dioxygen.

A flavohaemoprotein (FHP) from Ralstonia eutropha, obtained in a pure and active form, has been entrapped in a film of methyl cellulose on the electrode surface and gives a stable and reproducible electrochemical response at pH 7.00 when subject to cyclic voltammetry using a glassy carbon electrode. To our knowledge, no previous direct electrochemistry had been achieved with a bacterial flavohaemoglobin, which possess both a FAD and a haem. A single couple is observed which is assigned to the haem moiety of the protein, since the same result is obtained with a semi-apo form of the protein deprived of FAD (semi-apo FHP). The data collected were further confirmed by potentiometry with a platinum electrode, and the homogeneous electron transfer rate estimated by double potential step chronocoulometry at a bare glassy carbon electrode in the presence of methyl viologen (MV). The presence of FAD in the holoprotein is easily confirmed by UV-Vis spectrophotometry, but its expected electron relay role remains elusive. The protein activates the reduction of dioxygen by about 400 mV, the reduction current being proportional to the concentration of dioxygen up to 10% in volume in the gas mixture.

Bacterial Proteins↗
PubMed2004

Glass ionomer cement: evidence pointing to fluorine release in the form of monofluorophosphate in addition to fluoride ion.

The fluoride ion released from glass ionomer cements into water is reportedly, in part, complexed with other elements present in the cement. When measured using ion selective electrode potentiometry (ISE) a decomplexant TISAB IV (T) is used to convert all fluoride to F- ion which the ISE can detect. In this study, an additional decomplexing procedure (H) designed to hydrolyse fluorine covalently bonded to phosphorus in the monofluorophosphate (MFP) ion into F- was also used. The soluble products from three glass ionomers were analysed by both techniques (H & T). Five 1 x 10 mm discs were each immersed in 10 ml of de-ionised water. This was changed and 4 ml analysed by T and 4 ml by H at 1, 2, 3, 6, 10, 13, 17, 21, 24, 28, and 31 days. H was greater than T for 161 of the 165 pairs ( chi2=74.7, p=<0.001 ). The total cumulative F release H (in micromol/g cement) at 31 days for AH2 was 122.3, s.d. 30.8; LG26 44.0, s.d. 1.55; LG30 10.0, s.d. 3.15 as compared T results of 100.1, s.d. 31.1; 30.3, s.d. 1.92; 3.7, s.d. 1.36, respectively. In all three cases the H was significantly greater than T (matched pair 't' test with p=0.01 or less). H-T was show to have a very strong associative relationship with t1/2 (R2=0.98 or greater p<0.001 ). Evaluating the ratio of P:F in the cements in comparison with the ratio of additional F measured by H to that measured by T produced a relationship log[(H-T)/T]=0.28 x log[P/F]-0.45 with R2=0.999. It is concluded that glass ionomers release more fluorine than is detected by ISE using TISAB IV. If this F is in the form of MFP this may be released more completely into saliva than F as F-, release of which is substantially reduced by Ca2+, since calcium monofluorophosphate is more soluble than CaF2.

Electrochemistry↗
PubMed2005

Biosorption of cadmium by biomass of brown marine macroalgae.

Five different brown seaweeds, Bifurcaria bifurcata, Saccorhiza polyschides, Ascophyllum nodosum, Laminaria ochroleuca and Pelvetia caniculata were studied for their ability to remove cadmium from aqueous solution. Kinetics of cadmium adsorption by all the algae were relatively fast, with 90% of total adsorption occurring in less than 1h. These experiments could be accurately described by a pseudo-second-order rate equation, obtaining values between 1.66x10(-3) and 9.92x10(-3) g/mgmin for the sorption rate constant k. Several equilibrium adsorption isotherms were obtained for the quantitative description of cadmium uptake. The use of the Langmuir isotherm led to values between 64 and 95 mg/g for qmax and between 0.036 and 0.094 L/mg for b. The effect of pH on biosorption was also studied. Acid-base properties of algae were studied by potentiometry to determine pK values (from 3.54 to 3.98) and the total number of acid groups.

Adsorption↗
PubMed2006

Sequestration of biogenic amines by alginic and fulvic acids.

The interaction of natural (alginic and fulvic acids) and synthetic (polyacrylic acid 2.0 kDa) polyelectrolytes with some protonated polyamines [diamines: ethylendiamine, 1,4-diaminobutane (or putrescine), 1,5-diaminopentane (or cadaverine); triamines: N-(3-aminopropyl)-1,4-diaminobutane (or spermidine), diethylenetriamine; tetramine: N,N'-bis(3-aminopropyl)-1,4-diaminobutane (or spermine); pentamine: tetraethylene-pentamine; hexamine: pentaethylenehexamine] was studied at T=25 degrees C by potentiometry and calorimetry. Measurements were performed without supporting electrolyte, in order to avoid interference, and results were reported at I=0 mol L(-)(1). For all the systems, the formation of (am)L(2)H(i) species was found (am=amine; L=polyelectrolyte; i=1...4, depending on the amine considered). The stability of polyanion-polyammonium cation complexes is always significant, and for high-charged polycations, we observe a stability comparable to that of strong metal complexes. For example, by considering the formation reaction (am)H(i)+2L=(am)L(2)H(i) we found log K(i)=6.0, 6.5 and 10.8 for i=1, 2 and 3, respectively, in the system alginate-spermidine. Low and positive formation DeltaH(degrees) values indicate that the main contribution to the stability is entropic in nature. The sequestering ability of polyelectrolytes toward amines was modelled by a sigmoid Boltzman type equation. Some empirical relationships between stability, charges and DeltaG(degrees) and TDeltaS(degrees) are reported. Mean values per salt bridge of formation thermodynamic parameters (DeltaX(degrees) (n)) are DeltaG(degrees) (n)=-5.8+/-0.4, DeltaH degrees (n)=0.7+/-0.5 and TDeltaS(degrees) (n)=6.5+/-0.5 kJmol(-)(1) for all the systems studied in this work.

Alginates↗
PubMed2005

Evidence of two-step deprotonation of D-mannitol in aqueous solution.

Deprotonation of D-mannitol was studied in aqueous basic solutions by means of potentiometry and (13)C NMR spectroscopy. Two-step dissociation in the pH range from 12 to 13.8 was shown, and successive dissociation constants K(a1) and K(a2) were determined. In a solution with ionic strength I = 1.0 M (NaOH + NaNO(3)) pK(a1) = 13.1 +/- 0.1 and pK(a2) = 13.8 +/- 0.2. With increasing ionic strength from 0.75 to 3.0 M, both pK(a1) and pK(a2) values decrease. Deprotonation-induced chemical shifts in pH-variable (13)C NMR spectra show that the OH-groups next to internal carbon atoms C-3 and C-4 dissociate to a greater extent compared to OH-groups next to external carbon atoms C-1 and C-6.

Hydrogen-Ion Concentration↗
PubMed2005

Structure-property studies on the antioxidant activity of flavonoids present in diet.

The screening of natural flavonoids for their bioactivity as antioxidants is usually carried out by determinination of their profile as chain-breaking antioxidants, by the evaluation of their direct free radical-scavenging activity as hydrogen- or electron-donating compounds. Since this may not be the only mechanism underlying the antioxidant activity it is important to check the ability of these compounds to act as chelators of transition metal ions. Accordingly, in the present study the acidity constants of catechin and taxifolin, as well as the formation constants of the corresponding copper (II) complexes, were investigated by potentiometry and/or spectrophotometry. Moreover, a detailed quantitative examination of the coordination species formed is presented. In addition, the partition coefficients of both catechin and taxifolin in a biomimetic system (micelles) were determined, since these properties may also contribute to the antioxidant behavior of this type of compound. The log P values determined depend on the electrostatic interactions of the compounds with the differently charged micelles (the highest values were obtained for zwitterionic and cationic micelles). The prooxidant behavior of the compounds was assessed through the oxidation of 2'-deoxyguanosine, induced by a Fenton reaction, catalyzed by copper. The data obtained reveal that the flavonoids under study did not present prooxidant activity, in this particular system. The results obtained are evidence of a clear difference among the pKa, the complexation properties, and the lipophilicity of the flavonoids studied, which can partially explain their distinct antioxidant activity. The most stable geometries of the free compounds were determined by theoretical (ab initio) methods, in order to properly account for the electron correlation effects which occur in these systems, thus allowing a better interpretation of the experimental data.

Antioxidants↗