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pH Dependent (ph + dependent)
Selected AbstractsElectrochemical Preparation of Poly(acriflavine) Film-Modified Electrode and Its Electrolcatalytic Properties Towards NADH, Nitrite and Sulfur OxoanionsELECTROANALYSIS, Issue 9 2007Shen-Ming Chen Abstract Electrochemical polymerization of acriflavine (AF) was carried out onto glassy carbon electrodes (GCE) from the aqueous buffer solution containing 1.5×10,3,M AF monomer (pH,3.5) which produced a thin electrochemically active film. This is noted as poly(AF) film modified electrodes (PAF/GCE). This modified electrode was shown a stable reversible redox couple centered at +0.22,V in pH,3.5 buffer solutions. PAF/GCE was found to be more stable in acidic solutions and its formal potential was found to be pH dependent with a slope close to ,60,mV/pH. The electrochemical deposition kinetics of poly(AF) onto gold coated quartz crystal was studied by using electrochemical quartz crystal microbalance (EQCM) combined with cyclic voltammetry (CV). PAF/GCE was found be good mediator for electrochemical oxidation of reduced nicotinamide adenine dinucleotide (NADH) in pH,5 buffer solutions. The electrocatalytic oxidation of SO and electrocatalytic reduction of NO, SO and S2O were carried out at PAF/GCE electrode in acidic aqueous solutions. The electrocatalytic oxidation of NADH was also investigated by using amperometric method. [source] Solid State Electrochemical Oxidation Mechanisms Of Morin in Aqueous MediaELECTROANALYSIS, Issue 9 2005Patricia Janeiro Abstract The mechanism of electrochemical oxidation of morin has been studied using cyclic, differential pulse and square-wave voltammetry techniques in aqueous electrolyte with solid, insoluble morin hydrate mechanically transferred to a glassy carbon electrode surface, over a wide pH range. The oxidation mechanism proceeds in sequential steps, related with the hydroxyl groups in the three aromatic rings and the oxidation is pH dependent over part of the pH range the oxidation potentials are shifted to lower values with increasing pH. Oxidation of the 2,,4,dihydroxy moiety at the B ring of morin occurs first, at very low positive potentials, and is a one electron one proton reversible reaction. The hydroxyl groups oxidized at more positive potentials were shown to undergo an irreversible oxidation reaction. [source] Electrochemical Oxidation of QuercetinELECTROANALYSIS, Issue 22 2003Maria, Oliveira Brett Abstract The mechanism of electrochemical oxidation of quercetin on a glassy carbon electrode has been studied using cyclic, differential pulse and square-wave voltammetry at different pH. It proceeds in a cascade mechanism, related with the two catechol hydroxyl groups and the other three hydroxyl groups which all present electroactivity, and the oxidation is pH dependent. Quercetin also adsorbs strongly on the electrode surface; and the final oxidation product is not electroactive and blocks the electrode surface. The oxidation of the catechol 3,,4,-dihydroxyl electron-donating groups, occurs first, at very low positive potentials, and is a two electron two proton reversible reaction. The hydroxyl group oxidized next was shown to undergo an irreversible oxidation reaction, and this hydroxyl group can form a intermolecular hydrogen bond with the neighboring oxygen. The other two hydroxyl groups also have an electron donating effect and their oxidation is reversible. [source] Cadmium uptake by earthworms as related to the availability in the soil and the intestineENVIRONMENTAL TOXICOLOGY & CHEMISTRY, Issue 8 2001Leonard A. Oste Abstract The free metal concentration in the soil solution is often considered a key parameter for metal uptake by and toxicity to soft-bodied soil organisms. The equilibrium partitioning theory, which assumes a relationship between the contaminant concentration in pore water and the contaminant concentration in the body tissue, can be used to describe uptake by earthworms. This theory has proved useful for organic chemicals, but its applicability is less clear for metals. In this study, the Cd concentration in soil pore water (pw) was varied by increasing the soil pH by the addition of lime (Ca(OH)2) and by adding manganese oxide (MnO2), which has a high metal binding capacity. Both lime (0.135% w/w) and MnO2 (1% w/w) decreased [Cd2+]pw by a factor of 25, while CdWorm was reduced only by a factor of 1.3 in lime-treated soils and 2.5 in MnO2 -treated soils. Cadmium uptake was weakly related to the free metal concentration (R2adj = 0.66). Adding pH as an explanatory variable increased R2adj to 0.89, indicating that Cd uptake from pore water is pH dependent, which might be attributed to competition of protons and Cd at the surface of the earthworm body. However, previous earthworm experiments in reconstituted groundwater showed a conspicuously smaller pH dependency of Cd uptake. The differences in metal uptake between earthworms in lime- and MnO2 -treated soils are therefore more likely to reflect the predominance of pH-independent intestinal uptake of Cd. Equilibrating the soil with a solution of 0.01 M CaCl2 and 0.1 M triethanolamine (buffered at pH 7.2), simulating the conditions prevailing in the worm intestine, yielded free Cd concentrations that were closely (R2adj = 0.83) and linearly related to the Cd concentration in the earthworm tissue. [source] First evidence of catalytic mediation by phenolic compounds in the laccase-induced oxidation of lignin modelsFEBS JOURNAL, Issue 17 2003Francesca D'Acunzo The sulfonephthalein indicator, phenol red, exhibits an unusually slow rate of oxidation by laccase from Poliporus pinsitus, in spite of the fact that it is a phenol and therefore a natural substrate for this phenoloxidase enzyme. Nevertheless, after prolonged exposure to laccase (24 h) phenol red is oxidized by more than 90%. We found that phenol red, which can be oxidatively converted into a resonance-stabilized phenoxy radical, performs as a mediator in the laccase-catalyzed oxidation of a nonphenolic substrate (4-methoxybenzyl alcohol) and also of a hindered phenol (2,4,6-tri- tert -butylphenol). In particular, phenol red was found to be at least 10 times more efficient than 3-hydroxyanthranilate (a reported natural phenolic mediator of laccase) in the oxidation of 4-methoxybenzyl alcohol. Other phenols, which do not bear structural analogies to phenol red, underwent rapid degradation and did not perform as laccase mediators. On the other hand, several variously substituted sulfonephthaleins, of different pK2 values, mediated the laccase catalysis, the most efficient being dichlorophenol red, which has the lowest pK2 of the series. The mediating efficiency of phenol red and dichlorophenol red was found to be pH dependent, as was their oxidation Ep value (determined by cyclic voltammetry). We argue that the relative abundance of the phenoxy anion, which is easier to oxidize than the protonated phenol, may be one of the factors determining the efficiency of a phenolic mediator, together with its ability to form relatively stable oxidized intermediates that react with the desired substrate before being depleted in undesired routes. [source] Synthesis of pH dependent chitosan-EPI hydrogel films and their application for in vitro release of promethazine hydrochlorideJOURNAL OF APPLIED POLYMER SCIENCE, Issue 1 2008Yolda Abstract Chitosan-epichlorohydrin hydrogel films (ChitEPI) were synthesized by using chitosan in the presence of epichlorohydrin (EPI) as a crosslinking agent at various amounts. SEM, FTIR, TGA, and DSC analysis were conducted for the characterization of the hydrogels. The DSC measurements indicate that ChitEPI hydrogels did not exhibit better thermal stability when compared to chitosan. Swelling behavior of Chitosan-EPI hydrogel film is pH dependent and showed a reversible swelling behavior with a fast response. The hydrogels were used for in vitro release of promethazine hydrochloride (PHCl) in pH = 1.2 and pH = 7.4 phosphate buffer solutions (PBS). The release of PHCl synthesized from hydrogels at pH = 7.4 is quite low while at pH = 1.2, the highest value was observed as 49% for ChitEPI600. It has been also found that PHCl release from ChitEPI thin films is mainly controlled by diffusion control mechanism. ChitEPI hydrogels may be used for the delivery of drug in stomach and gastrointestinal tract. © 2008 Wiley Periodicals, Inc. J Appl Polym Sci, 2008 [source] Separation of cobalt and nickel from acidic sulfate solutions using mixtures of di(2-ethylhexyl)phosphoric acid (DP-8R) and hydroxyoxime (ACORGA M5640)JOURNAL OF CHEMICAL TECHNOLOGY & BIOTECHNOLOGY, Issue 5 2004Arisbel Cerpa Abstract DP-8R and ACORGA M5640 extractants diluted in Exxsol D100 were used to co-extract cobalt and nickel from aqueous acidic sulfate media. The influences of equilibration time, temperature, equilibrium pH and reagent concentrations on the extraction of both metals have been studied. It was observed that both cobalt and nickel extraction are slightly sensitive to temperature but are pH dependent. Metal extraction equilibria are reached within about 5 min contact time. In addition, cobalt extraction depends on the extractant concentration in the organic phase. For a solution containing 0.5 g dm,3 each of cobalt and nickel and an initial pH of 4.1, conditions were established for the co-extraction of both metals and selective stripping (with H2SO4) of cobalt and nickel. Using the appropriate reagent concentrations the yield (extraction stage) for both metals exceeded 90%, and stripping of cobalt and nickel was almost quantitative. Copyright © 2004 Society of Chemical Industry [source] Relative in vitro efficacy of the phosphate binders lanthanum carbonate and sevelamer hydrochlorideJOURNAL OF PHARMACEUTICAL SCIENCES, Issue 10 2007Valerie Autissier Abstract The high tablet burden and poor compliance associated with phosphate-binding drugs has led to a search for more potent agents. In vitro -binding studies were performed on the recently introduced binder, lanthanum carbonate (LC; Fosrenol®), to compare its phosphate-binding affinity with sevelamer hydrochloride (SH; RenagelÔ). Langmuir equilibrium binding affinities (K1) for LC and SH were established using different phosphorus (5,100 mM) and binder (134,670 mg per 50 mL) concentrations at pH 3,7, with or without salts of bile acids present (30 mM). At all pH levels, LC had a higher binding affinity for phosphate than SH. For LC, K1 was 6.1,±,1.0 mM,1 and was independent of pH. For SH, K1 was pH dependent, being 1.5,±,0.8 mM,1 at pH 5,7 and 0.025,±,0.002 mM,1 at pH 3, that is, >200 times lower than for LC. In the presence of 30 mM bile salts, SH lost 50% of its phosphate, whereas no displacement of phosphate occurred for LC. These findings indicate that LC binds phosphate more effectively than SH across the pH range encountered in the gastrointestinal tract, and has a lower propensity for bound phosphate to be displaced by competing anions in the intestine. © 2007 Wiley-Liss, Inc. and the American Pharmacists Association J Pharm Sci 96: 2818,2827, 2007 [source] Site-specific contribution of proton-coupled folate transporter/haem carrier protein 1 in the intestinal absorption of methotrexate in ratsJOURNAL OF PHARMACY AND PHARMACOLOGY: AN INTERNATI ONAL JOURNAL OF PHARMACEUTICAL SCIENCE, Issue 7 2009Tomoharu Yokooji Abstract Objectives Methotrexate is reportedly a substrate for proton-coupled folate transporter/haem carrier protein 1 (PCFT/HCP1) and reduced folate carrier 1 (RFC1). In this study, we examined the contribution of PCFT/HCP1 and RFC1 in the intestinal absorption of methotrexate in rats. Methods Western blot analysis was carried out to evaluate the protein levels of PCFT/HCP1 and multidrug resistance-associated protein 2 in brush-border membrane of rat small intestine. Mucosal uptake of methotrexate was studied in the rat everted small intestine and an in-situ intestinal perfusion study of methotrexate was also carried out in rats. Key findings In transport studies using everted intestine, the mucosal methotrexate influx rate in proximal intestine at pH 5.5 was significantly greater than that at pH 7.4. Coadministration of folate or its analogues, such as folinate and 5-methyltetrahydrofolate, substrates for both PCFT/HCP1 and RFC1, significantly suppressed the methotrexate influx at pH 5.5, whereas thiamine pyrophosphate, an inhibitor for RFC1 alone, exerted no significant effect. Western blot analysis showed higher PCFT/HCP1 expression in proximal than distal small intestine. In distal small intestine, methotrexate influx rate was low and was not pH dependent. Also, folate and its analogues exerted no significant effect on methotrexate absorption. Conclusions Based on the present and our previous results, the site-specific contributions of various transporters including PCFT/HCP1 in methotrexate intestinal absorption were discussed. The variation in luminal pH and the involvement of multiple transporters in methotrexate absorption may cause variation in oral bioavailability among patients. [source] Oral peptide delivery: in-vitro evaluation of thiolated alginate/poly(acrylic acid) microparticlesJOURNAL OF PHARMACY AND PHARMACOLOGY: AN INTERNATI ONAL JOURNAL OF PHARMACEUTICAL SCIENCE, Issue 9 2007Alexander Greimel ABSTRACT The purpose of this study was to develop an oral thiomer-based microparticulate delivery system for insulin by ionic gelation. The microparticulate matrix consisted of either poly(acrylic acid)-cysteine (PAA-Cys) and alginate-cysteine (Alg-Cys) or the corresponding unmodified polymers (PAA, Alg). Two different viscosities of alginates were provided for the study, low and medium. Three different types of microparticles were prepared via ionic gelation with calcium (Alg, AlgPAA and AlgPAA-Cys) and their different properties evaluated in-vitro (particle size and shape, drug loading and release profile, swelling and stability). The mean particle size of all formulations ranged from 400 to 600 ,m, revealing the lowest for thiolated microparticles. SEM micrographs showed different morphological profiles for the three different types of microparticles. Encapsulation efficiency of insulin increased within the following rank order: Alg (15%) < AlgPAA (40%) < AlgPAA-Cys (65%). Alginate and AlgPAA microparticles displayed a burst release after 30 min, whereas the thiolated particles achieved a controlled release of insulin over 3 h. The swelling ratio was pH dependent: in simulated intestinal fluid microparticles exhibited a much higher water uptake compared with simulated gastric fluid. Due to the formation of intraparticulate disulfide bonds during the preparation process, thiolated particles revealed a higher stability. It was also observed that the viscosity of the two alginates used had no influence on the properties of the particles. According to these results AlgPAA-Cys microparticles obtained by ionic gelation and stabilized via disulfide bonds might be an alternative tool for the oral administration of therapeutic peptides. [source] Raman spectroscopy of hydrotalcites with phosphate in the interlayer: implications for the removal of phosphate from waterJOURNAL OF RAMAN SPECTROSCOPY, Issue 7 2006Ray L. Frost Abstract Hydrotalcites with phosphate in the interlayer were prepared at different pH values. At pH > 11.0 (PO4)3, was the intercalated ionic species, whereas at pH < 11.0 a mixture of (PO4)3, and (HPO4)2, ions was intercalated. Powder X-ray diffraction shows that the hydrotalcite formed at pH 9.5 is poorly diffracting with a d-spacing of 11.9 Å; whereas the d(003) spacing for the phosphate interlayered hydrotalcite formed at pH 11.9 and 12.5 was 8.0 and 7.9 Å respectively. The addition of a thermally activated ZnAl-HT to a phosphate solution resulted in the uptake of the phosphate and the reformation of the hydrotalcite. Raman spectroscopy of the phosphate interlayered hydrotalcites shows that the interlayered anion is pH dependent and only above pH 11.9 is the orthophosphate anion intercalated. At lower pH, the monohydrogen phosphate anion is intercalated. Raman spectroscopy shows that upon addition of the thermally activated hydrotalcite to an aqueous phosphate solution, results in the uptake of phosphate anion from the solution. Copyright © 2006 John Wiley & Sons, Ltd. [source] Anti-mitotic activity of colchicine and the structural basis for its interaction with tubulinMEDICINAL RESEARCH REVIEWS, Issue 1 2008Bhabatarak Bhattacharyya Abstract In this review, an attempt has been made to throw light on the mechanism of action of colchicine and its different analogs as anti-cancer agents. Colchicine interacts with tubulin and perturbs the assembly dynamics of microtubules. Though its use has been limited because of its toxicity, colchicine can still be used as a lead compound for the generation of potent anti-cancer drugs. Colchicine binds to tubulin in a poorly reversible manner with high activation energy. The binding interaction is favored entropically. In contrast, binding of its simple analogs AC or DAAC is enthalpically favored and commences with comparatively low activation energy. Colchicine,tubulin interaction, which is normally pH dependent, has been found to be independent of pH in the presence of microtubule-associated proteins, salts or upon cleavage of carboxy termini of tubulin. Biphasic kinetics of colchicines,tubulin interaction has been explained in light of the variation in the residues around the drug-binding site on , -tubulin. Using the crystal structure of the tubulin,DAMAcolchicine complex, a detailed discussion on the pharmacophore concept that explains the variation of affinity for different colchicine site inhibitors (CSI) has been discussed. © 2007 Wiley Periodicals, Inc. Med Res Rev, 28, No. 1, 155,183, 2008 [source] The effect of experimental conditions on the detection of spermine in cell extracts and tissuesNMR IN BIOMEDICINE, Issue 2 2010Nicholas G. Spencer Abstract The aim of this work was to investigate the effect of experimental conditions on the visibility of polyamines. In solution the chemical shift of the three groups of peaks (at approximately 1.8, 2.1 and 3.1,ppm) were found to be pH dependent. Relaxation times in aqueous solution at pH 7.0, 298,K and 11.74,T were measured to be: putrescine (T1,=,2.49,s, T2,=,2.07,s), spermidine (T1,=,1.27,s, T2,=,1.05,s) and spermine (T1,=,1.02,s, T2,=,0.82,s). Simple spin-echo sequences could not be used to measure T2 as the spins also experience phase evolution from homonuclear coupling which imposes a modulation on the T2 decay curve. This modulation is eliminated by using CPMG sequences with an echo spacing of <500,µs. Relaxation times for spermine in solution in presence of metal ions and protein showed that metal ions had little effect on T2; however, addition of 15,mg/ml bovine serum albumin reduced T2 of spermine (0.41,s at 298,K and 0.19,s at 277,K) but was not as short as the T2 of the polyamine peak in prostatic tissue (0.03,s at 277,K). The MR visibility of polyamines in prostate cell extracts, PC-3 xenograft (intact as well as extracted) and intact human prostatic tissues were investigated. Polyamines were not detected in methanol/chloroform extracts, but were visible in perchloric acid extracts of prostate tumour cells. No polyamines were detected in the HR MAS spectra of three samples of whole PC-3 xenograft tissue studied. In summary, the chemical shift of polyamine species is pH dependent, while protein binding causes peak broadening and reduction in T2. Perchloric acid extraction improves visibility of intracellular polyamines, but whole tissue polyamines are not seen in xenografts without epithelial/ ductal structure. Copyright © 2009 John Wiley & Sons, Ltd. [source] Copper binding to octarepeat peptides of the prion protein monitored by mass spectrometryPROTEIN SCIENCE, Issue 2 2000Randy M. Whittal Abstract Electrospray ionization mass spectrometry (ESI-MS) was used to measure the binding of Cu2+ ions to synthetic peptides corresponding to sections of the sequence of the mature prion protein (PrP). ESI-MS demonstrates that Cu2+ is unique among divalent metal ions in binding to PrP and defines the location of the major Cu2+ binding site as the octarepeat region in the N-terminal domain, containing multiple copies of the repeat ProHisGlyGlyGlyTrpGlyGln. The stoichiometries of the complexes measured directly by ESI-MS are pH dependent: a peptide containing four octarepeats chelates two Cu2+ ions at pH 6 but four at pH 7.4. At the higher pH, the binding of multiple Cu2+ ions occurs with a high degree of cooperativity for peptides C-terminally extended to incorporate a fifth histidine. Dissociation constants for each Cu2+ ion binding to the octarepeat peptides, reported here for the first time, are mostly in the low micromolar range; for the addition of the third and fourth Cu2+ ions to the extended peptides at pH 7.4, KD's are <100 nm. n-terminal acetylation of the peptides caused some reduction in the stoichiometry of binding at both ph's. cu2+ also binds to a peptide corresponding to the extreme N-terminus of PrP that precedes the octarepeats, arguing that this region of the sequence may also make a contribution to the Cu2+ complexation. Although the structure of the four-octarepeat peptide is not affected by pH changes in the absence of Cu2+, as judged by circular dichroism, Cu2+ binding induces a modest change at pH 6 and a major structural perturbation at pH 7.4. It is possible that PrP functions as a Cu2+ transporter by binding Cu2+ ions from the extracellular medium under physiologic conditions and then releasing some or all of this metal upon exposure to acidic pH in endosomes or secondary lysosomes. [source] Glutamine deamidation of a recombinant monoclonal antibodyRAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 24 2008Hongcheng Liu Deamidation of glutamine (Gln) proceeds at a much slower rate than deamidation of asparagine (Asn) residues at peptide level. However, deamidation of Gln residues in native proteins may occur faster because of the impact of protein structure and thus plays a significant role in affecting protein stability. Gln deamidation of a recombinant monoclonal IgG1 antibody was investigated in the current study. Deamidation was determined by a molecular weight increase of 1,Da, a retention time shift on reversed-phase chromatography and tandem mass spectrometric (MS/MS) analysis of the peptides. As expected, Gln residues at different locations in the three-dimensional structure had different susceptibilities to deamidation. Gln deamidation was highly pH dependent with the highest level detected in the sample incubated at pH 9, and lowest level at pH 6 in the pH range from 5 to 9. The detection of significant levels of Gln deamidation suggested that it may play an important role in affecting heterogeneity and stability of recombinant monoclonal antibodies. Copyright © 2008 John Wiley & Sons, Ltd. [source] Iron, Manganese and Copper Equilibria with Wood Fibres in Single Salt Aqueous SuspensionsTHE CANADIAN JOURNAL OF CHEMICAL ENGINEERING, Issue 3 2005Robin Susilo Abstract Ions of Fe, Mn and Cu were introduced into suspensions of protonated or metal-free fibres and the equilibrium concentration of each metal in the fibre and surrounding solution was measured. The results were compared with the Donnan equilibrium model. Mn and Cu concentrations on the fibres were found to be pH dependent and in agreement with the model. An increased amount of Fe on the Kraft pulp fibres was found and attributed to iron containing precipitates trapped within the fibres. Precipitates in mechanical pulps had a very small amount (1-2 wt%) of iron. Des ions de Fe, Mn et Cu ont été introduits dans des suspensions de fibres protonatées ou « sans métaux » et la concentration d'équilibre de chaque métal dans la fibre et la solution environnante a été mesurée. Les résultats ont été comparés avec le modèle d'équilibre de Donnan. On a trouvé que les concentrations de Mn et Cu sur les fibres étaient dépendantes du pH et en accord avec le modèle. Une quantité accrue de Fe a été trouvée sur les fibres de pâte kraft et attribuée aux précipitats contenant du fer retenus dans les fibres. Les précipitats dans les pâtes mécaniques ont une très petite quantité (1-2% en poids) de fer. [source] Development of Cellulose-DNA ImmunoadsorbentARTIFICIAL ORGANS, Issue 2 2002Deling Kong Abstract: The aim of this study was to prepare a DNA immunoadsorbent for the specific, extracorporeal removal of anti-DNA antibodies from the blood of patients with systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA). Two kinds of cellulose beads were applied as a carrier. Calf thymus DNA was covalently coupled to the carrier using the epichlorohydrin method. Efforts were focused on optimization of conditions for activation and coupling, trying to couple as much DNA as possible to a certain amount of carrier. It was found that the activation level increased with the increase of NaOH concentration and the amount of epichlorohydrin used. The mole of epichlorohydrin must be in excess of that of NaOH because excess NaOH could react further with the epoxy groups in the beads resulting in a decrease of activation level. High activation level could be obtained in a medium of 3.0 M NaOH. The DNA coupling was found to be mainly temperature and pH dependent. Using 0.1 M Tris-HCl buffer, pH 8 at a temperature of 50,90°C, more than 3 mg of DNA could be coupled to 1 ml of wet beads. Prolonging the coupling reaction under 50°C to 72 h resulted in the same coupling capacity as that obtained under 90°C. To evaluate the adsorption ability for anti-DNA of this immunoadsorbent, batch and circulation tests were applied using SLE patient plasma. The immunoadsorbents showed excellent adsorption capacity, especially the cellulose with smaller size (200,300 ,m). The incubation of 20 ml of patient's plasma with 1 ml of adsorbent resulted in an 80% decline in the anti-DNA antibody level. In the circulation tests, 30 ml of plasma was circulated through a column containing 3 ml of adsorbent. The maximum decline in anti-DNA level, 80%, was obtained after 60 min. Such high adsorption capacity and high adsorption rate suggest this immunoadsorbent may be used for treatment. For comparison, 1,4-butanediol diglycidyl ether activation method and other DNA sources were tested with the same protocol. [source] Comparison of ceftibuten transport across Caco-2 cells and rat jejunum mounted on modified ussing chambersBIOPHARMACEUTICS AND DRUG DISPOSITION, Issue 7 2003R.M. Menon Abstract Ceftibuten uptake into Caco-2 cells and intestinal brush border membrane vesicles is mediated by the dipeptide transport system (PEPT1). The apical to basolateral transport characteristics of ceftibuten across Caco-2 cells and rat jejunum mounted on a modified Ussing chamber was examined. Mannitol was used as a paracellular marker along with trans-epithelial electrical resistance (TEER) for monitoring tight junction permeability. Transport across Caco-2 cells and rat jejunum mounted on a modified Ussing chamber was linear across the concentration range 0.25,10 mm. The net flux of mannitol and ceftibuten was higher across rat jejunum compared with Caco-2 cells. At a donor concentration of 0.25 mm, ceftibuten transport across Caco-2 cells was found to be pH dependent. Glycyl proline, a dipeptide, and 2,4- dinitrophenol, an energy poison, caused a reduction in the permeability of 0.25 mm ceftibuten across Caco-2 cells. Benzoic acid and adipic acid also inhibited transcellular transport of ceftibuten. At a donor concentration of 0.25 mm, passive paracellular transport accounts for about 60% and the active carrier mediated mechanism accounts for about 40% of ceftibuten transport across Caco-2 cells. None of the inhibitors however, had a significant effect on ceftibuten transport across rat jejunum mounted on a modified Ussing chamber at a donor concentration of 0.25 mm. In the concentration range 0.25,10 mm, ceftibuten is predominantly transported by paracellular mechanisms across rat jejunum and a mixture of active and passive transport across Caco-2 cells. Copyright © 2003 John Wiley & Sons, Ltd. [source] Characterization of copper binding to the peptide amyloid-,(1,16) associated with Alzheimer's diseaseBIOPOLYMERS, Issue 1 2006Qing-Feng Ma Abstract Amyloid-, peptide (A,) is the principal constituent of plaques associated with Alzheimer's disease (AD) and is thought to be responsible for the neurotoxicity associated with the disease. Copper binding to A, has been hypothesized to play an important role in the neruotoxicity of A, and free radical damage, and Cu2+ chelators represent a possible therapy for AD. However, many properties of copper binding to A, have not been elucidated clearly, and the location of copper binding sites on A, is also in controversy. Here we have used a range of spectroscopic techniques to characterize the coordination of Cu2+ to A,(1,16) in solution. Electrospray ionization mass spectrometry shows that copper binds to A,(1,16) at pH 6.0 and 7.0. The mode of copper binding is highly pH dependent. Circular dichroism results indicate that copper chelation causes a structural transition of A,(1,16). UV-visible absorption spectra suggest that three nitrogen donor ligands and one oxygen donor ligand (3N1O) in A,(1,16) may form a type II square-planar coordination geometry with Cu2+. By means of fluorescence spectroscopy, competition studies with glycine and L -histidine show that copper binds to A,(1,16) with an affinity of Ka , 107M,1 at pH 7.8. Besides His6, His13, and His14, Tyr10 is also involved in the coordination of A,(1,16) with Cu2+, which is supported by 1H NMR and UV-visible absorption spectra. Evidence for the link between Cu2+ and AD is growing, and this work has made a significant contribution to understanding the mode of copper binding to A,(1,16) in solution. © 2006 Wiley Periodicals, Inc. Biopolymers 83: 20,31, 2006 This article was originally published online as an accepted preprint. The "Published Online" date corresponds to the preprint version. You can request a copy of the preprint by emailing the Biopolymers editorial office at biopolymers@wiley.com [source] Spectroelectrochemical and Computational Studies on the Mechanism of Hypoxia Selectivity of Copper RadiopharmaceuticalsCHEMISTRY - A EUROPEAN JOURNAL, Issue 19 2008Jason Abstract Detailed chemical, spectroelectrochemical and computational studies have been used to investigate the mechanism of hypoxia selectivity of a range of copper radiopharmaceuticals. A revised mechanism involving a delicate balance between cellular uptake, intracellular reduction, reoxidation, protonation and ligand dissociation is proposed. This mechanism accounts for observed differences in the reported cellular uptake and washout of related copper bis(thiosemicarbazonato) complexes. Three copper and zinc complexes have been characterised by X-ray crystallography and the redox chemistry of a series of copper complexes has been investigated by using electronic absorption and EPR spectroelectrochemistry. Time-dependent density functional theory (TD-DFT) calculations have also been used to probe the electronic structures of intermediate species and assign the electronic absorption spectra. DFT calculations also show that one-electron oxidation is ligand-based, leading to the formation of cationic triplet species. In the absence of protons, metal-centred one-electron reduction gives the reduced anionic copper(I) species, [CuIATSM],, and for the first time it is shown that molecular oxygen can reoxidise this anion to give the neutral, lipophilic parent complexes, which can wash out of cells. The electrochemistry is pH dependent and in the presence of stronger acids both chemical and electrochemical reduction leads to quantitative and rapid dissociation of copper(I) ions from the mono- or diprotonated complexes, [CuIATSMH] and [CuIATSMH2]+. In addition, a range of protonated intermediate species have been identified at lower acid concentrations. The one-electron reduction potential, rate of reoxidation of the copper(I) anionic species and ease of protonation are dependent on the structure of the ligand, which also governs their observed behaviour in vivo. [source] Mannose binding lectin and C3 act as recognition molecules for infectious agents in the vaginaCLINICAL & EXPERIMENTAL IMMUNOLOGY, Issue 1 2005V. Pellis Summary In our study we examined the early complement components in patients with bacterial vaginosis (BV), vulvovaginal candidiasis (VVC) and in healthy controls. The levels of C1q, mannose-binding lectin (MBL) and C3 were measured by ELISA in the cervicovaginal lavage (CVL) from gynaecological patients and controls. No significant differences were observed in the levels of these proteins in the three study groups. Immunofluorescence analysis of the clue cells and Candida hyphae from BV and VVC patients for surface-bound complement components showed the presence of C3, while C1q was undetectable. MBL was revealed on clue cells but not on Candida. Binding of MBL to Candida, grown or cytocentrifuged from the CVL of VVC patients, was found to be pH dependent and occurred between pH 4·5 and pH 5·5. In conclusion, we demonstrated that MBL and C3 present in the vaginal cavity act as recognition molecules for infectious agents that colonize the cervicovaginal mucosa. Our finding that MBL, but not C1q, binds to bacteria and fungi in vagina suggests that the lectin and classical pathways of complement activation may play a different role in immune defence in the female genital tract. [source] | |||||||||||||||||||||||||