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Spectrophotometry
Kinds of Spectrophotometry Selected AbstractsComputerized Color Formulation for African-Canadian People Requiring Facial Prostheses: A Pilot StudyJOURNAL OF PROSTHODONTICS, Issue 4 2008FETC, MIMPT, MPhil, T.J. Coward PhD Abstract Purpose: The aim of this study was to investigate the effectiveness of spectrophotometry and a computerized color formulation system to predict pigment formulas for color mixing silicone elastomer to match the skin color of African-Canadian people. Materials and Methods: In a prospective study, reflectance spectrophotometery was used to measure the skin color of 19 African-Canadian subjects. The spectral data for each subject was used in a computerized color formulation system to predict colorants required to mix silicone elastomer to match each subject's skin color. Delta-E values were recorded for each silicone sample in comparison to the subject's skin measurement. An analysis of variance was used to determine significance among variables, and a Tukey HSD post hoc test was used to assess paired comparisons. Results: Delta-E decreased with iterative mixes of colored silicone for each subject, and pigment loading increased with iterative mixes. Delta-E values for the third iterative mix (fourth and final sample) ranged between 1.49 and 8.82. Conclusion: Spectrophotometry and computerized color formulation provide a foundation in the color matching procedure for facial prostheses that offers objectivity to an otherwise subjective task. Through further study of spectrophotometry and computerized color formulation, and with the development of pigment databases appropriate for the African-Canadian population, it may be possible to establish a precise and repeatable color matching system that predicts required colorants and controls metamerism. [source] Straightforward Determination of the Degree of N -Acetylation of Chitosan by Means of First-Derivative UV SpectrophotometryMACROMOLECULAR CHEMISTRY AND PHYSICS, Issue 14 2008Ricardo M. P. da Silva Abstract First-derivative UV spectrophotometry is shown to be a reliable method for the determination of the degree of N- acetylation of chitosan samples. A mathematical expression is derived that allows to determine the DA directly from the mass concentration of a chitosan solution and the first derivative of its UV spectrum at 202 nm, thus eliminating the need for empiric correction curves for highly deacetylated samples. A procedure is proposed for the accurate mass determination of the hygroscopic chitosan. The proposed approach facilitates the routine determination of the DA, especially when using potent multiwell microplate readers, which allow hundreds of samples to be measured in just a few minutes. [source] Comparison of Nonmetal and Metal Hydrophilic Photosensitizer, ATX-S10 (Na) and ATN-2, Binding with Human Serum Proteins Using Spectrophotometry,PHOTOCHEMISTRY & PHOTOBIOLOGY, Issue 2 2004M. Yamaguchi ABSTRACT Intermolecular interactions of human serum proteins with a hydrophilic nonmetalloporphyrin, 13,17-bis(1-carboxy-propionyl)carbomoylethyl-8-ethenyl-2-hydroxy-3-hydroxy-iminoethylidene-2,7,12,18-tetramethylporphyrin sodium salt (ATX-S10 (Na)), or a hydrophilic gallium-metalloporphyrin, diethylenetriamine pentaacetic acid ester of 2-[1-(2-hydroxyethoxy)ethyl]-4-vinyl-deuteroporphyrin (IX) Ga complex (ATN-2), were investigated using spectrophotometry. ATX-S10 (Na) caused a bathochromic shift with albumin, high-density lipoprotein and low-density lipoprotein, but little or no shift was observed with hemopexin, transferrin and immuno-globulin G. In contrast, ATN-2 displayed a bathochromic shift only with hemopexin. These results suggest that the association energy of ATX-S10 (Na) with albumin might be slightly greater than that with lipoproteins and that of ATN-2 with hemopexin might be greater than that with other serum proteins. [source] Solid Phase Extraction of Thallium(III) on Micro Crystalline Naphthalene Modified with N,N, -Bis(3-methylsalicylidene)- ortho -phenylenediamine and Determination by SpectrophotometryCHINESE JOURNAL OF CHEMISTRY, Issue 10 2008Ali MOGHIMI Abstract A novel, simple, sensitive and effective method has been developed for preconcentration of thallium on N,N, -bis(3-methylsalicylidene)- ortho -phenylenediamine (MSOPD) adsorbent in a pH range 5.0,10.0, prior to its spectrophotometric determination, based on the oxidation of bromopyrogallol red at ,=520 nm. This method makes it possible to quantitize thallium in a range of 3.6×10,9 to 2.0×10,5 mol/L, with a detection limit (S/N=3) of 1.42×10,9 mol/L. This procedure has been successfully applied to determine the ultra trace levels of thallium in the environmental samples, free from the interference of some diverse ions. The precision, expressed as relative standard deviation of three measurements, is better than 2.9%. [source] Methods for detecting and identifying retinoids in tissueDEVELOPMENTAL NEUROBIOLOGY, Issue 7 2006Thomas E. Gundersen Abstract Methods for retinoid analysis in tissue include direct spectrophotometry or fluorometry and retinoid responsive reporter constructs in the form of cell reporter assays or transgenic reporter animals, but chromatographic methods dominate and posses several superior features in quantitative analysis. The multitude of extraction protocols used can coarsely be divided into manual liquid-liquid extraction protocols and semi- or fully automated solid phase extraction-based protocols. Liquid chromatographic separation in reversed phase dominates although normal phase is also used. Detection is mainly performed with UV detectors although electrochemical and fluorescence detection is also used. Mass spectrometry in combination with LC is more often used in retinoid analysis and is likely to dominate in the future. © 2006 Wiley Periodicals, Inc. J Neurobiol 66: 631,644, 2006 [source] Simultaneous determination of metronidazole and spiramycin in bulk powder and in tablets using different spectrophotometric techniquesDRUG TESTING AND ANALYSIS, Issue 1 2010Fatma I. Khattab Abstract Metronidazole (MZ) is an anti-infective drug used in the treatment of anaerobic bacterial and protozoa infections in humans. It is also used as a vetinary antiparasitic drug. Spiramycin (SP) is a medium-spectrum antibiotic with high effectiveness against Gram-positive bacteria. Three simple, sensitive, selective and precise spectrophotometric methods were developed and validated for the simultaneous determination of MZ and SP in their pure form and in pharmaceutical formulations. In methods A and B, MZ was determined by the application of direct spectrophotometry and by measuring its zero-order (D0) absorption spectra at its ,max = 311 nm. In method A, SP was determined by the application of first derivative spectrophotometry (D1) and by measuring the amplitude at 218.3 nm. In method B, the first derivative of the ratio spectra (DD1) was applied, and SP was determined by measuring the peak amplitude at 245.6 nm. Method C entailed mean centring of the ratio spectra (MCR), which allows the determination of both MZ and SP. The methods developed were used for the determination of MZ and SP over a concentration range of 5,25 µg ml,1. The proposed methods were used to determine both drugs in their pure, powdered forms with mean percentage recoveries of 100.16 ± 0.73 for MZ in methods A and B, 101.10 ± 0.90 in method C, 100.09 ± 0.70, 100.02 ± 0.88 and 100.49 ± 1.26 for SP in methods A, B and C, respectively. The proposed methods were proved using laboratory-prepared mixtures of the two drugs and were successfully applied to the analysis of MZ and SP in tablet formulation without any interference from each other or from the excipients. The results obtained by applying the proposed methods were compared statistically with a reported HPLC method and no significant difference was observed between these methods regarding both accuracy and precision. Copyright © 2010 John Wiley & Sons, Ltd. [source] Effect of Enzyme and Cofactor Immobilization on the Response of Ethanol Oxidation in Zirconium Phosphate Modified BiosensorsELECTROANALYSIS, Issue 10 2010Mitk'El Abstract Two different self-contained ethanol amperometric biosensors incorporating layered [Ru(phend)2bpy]2+ -intercalated zirconium phosphate (ZrP) as the mediator as well as yeast -alcohol dehydrogenase (y- ADH) and its cofactor nicotinamide adenine dinucleotide (NAD+) were constructed to improve upon a design previously reported where only this mediator was immobilized in the surface of a modified electrode. In the first biosensor, a [Ru(phend)2bpy]2+ -intercalated ZrP modified carbon paste electrode (CPE) was improved by immobilizing in its surface both y- ADH and NAD+ using quaternized Nafion membrane. In the second biosensor, a glassy carbon electrode was modified with [Ru(phend)2bpy]2+ -intercalated ZrP, y- ADH, and NAD+ using Nafion as the holding matrix. Calibration plots for ethanol sensing were constructed in the presence and absence of ZrP. In the absence of ZrP in the surface of the modified glassy carbon electrode, leaching of ADH was observed as detected by UV-vis spectrophotometry. Ethanol sensing was also tested in the presence and absence of ascorbate to measure the selectivity of the sensor for ethanol. These two ethanol biosensors were compared to a previously reported one where the y -ADH and the NAD+ were in solution, not immobilized. [source] Voltammetric Investigation of Zinc Release from Metallothioneins Modulated by the Glutathione Redox Couple and Separated with a Porous MembraneELECTROANALYSIS, Issue 20 2008Lin Liu Abstract Glutathione (GSH), in addition to serving as a redox buffer in cellular environment, has been suggested as a modulator in metal regulation and homeostasis by metallothioneins (MTs). The interactions of MTs with both GSH and its oxidized form GSSG have been shown to govern the direction of metal transfer. Common methods for the determination of zinc release from MTs modulated by GSH/GSSG either involve radioactive species or enzymes or are labor-intensive. In this study, upon separation of Zn2+ from the reaction mixture of MTs and GSH with a centrifugal filter membrane, differential pulse voltammetry (DPV) was used for the Zn2+ quantification. The same approach is extended to the studies of metal transfer between Zn7MT with a GSH/GSSG mixture and that between Zn7MT with GSSG. The concomitant conversion between the free thiol and disulfide bonds was confirmed with UV-vis spectrophotometry. The results demonstrate that GSSG, GSH, and the GSH/GSSG mixture all modulate zinc release from Zn7MT. The percentage of zinc release increases in the order of GSH, GSSG, and the GSH/GSSG mixture. The new approach is demonstrated to be well suited for investigation of redox regulation of MT and its reaction with zinc-containing enzymes. [source] FIA Determination of Paracetamol in Pharmaceutical Drugs by Using Gold Electrodes Modified with a 3-Mercaptopropionic Acid MonolayerELECTROANALYSIS, Issue 9 2006Valber Abstract A flow injection analysis (FIA) method for the determination of paracetamol in pharmaceutical drugs using a gold electrode modified with a self-assembled monolayer (SAM) of 3-mercaptopropionic acid is described. At optimized experimental conditions the dynamic concentration range was 0.15 to 15.0,mg L,1 with a detection limit of 0.2,,g mL,1 (S/N=3). The repeatability of current responses for injections of 10,,mol L,1 paracetamol was evaluated to be 3.2% (n=30) and the analytical frequency was 180,h,1. The lifetime of the modified electrode was found to be 15 days. The results obtained by using the proposed amperometric method for paracetamol determination in four different drug samples compared well with those found by spectrophotometry. [source] Electropolymerized Pyrrole-Substituted Manganese Phthalocyanine Films for the Electroassisted Biomimetic Catalytic Reduction of Molecular OxygenELECTROANALYSIS, Issue 2 2005Nazaré, Pereira Rodrigues Abstract We report for the first time on the electroassisted biomimetic activation of molecular oxygen by a newly prepared electropolymerized polypyrrole-manganese phthalocyanine film. The prepared films and their intervention in the electroassisted catalytic reduction of molecular oxygen were analyzed by cyclic voltammetry and UV-visible spectrophotometry on optically transparent electrodes. The obtained results demonstrate the probable existence of the key-steps responsible for the suggested formation of the highly reactive manganese oxo intermediate. [source] Batch Injection Analysis: An Almost Unexplored Powerful ToolELECTROANALYSIS, Issue 7 2004Maria Abstract The main purpose of this review is to report the development and potentialities of batch injection analysis (BIA) and call the attention of researchers about the power of this (until now) almost unexplored tool. The text focuses on the concepts and potentialities of BIA combined with other techniques for analytical purposes. The association of BIA with amperometry, potentiometry, voltammetry, calorimetry, fluorescence and spectrophotometry is presented and important aspects related with each technique are discussed. Some aspects comparing the similarities and differences between BIA, FIA and wall-jet are presented along the text. [source] Factors affecting biodegradation of 2-chlorophenol by Alcaligenes sp. in aerobic reactorsENVIRONMENTAL TOXICOLOGY, Issue 4 2001A. Gallego Abstract The influence of variations in carbon source concentration, cell inocula, pH, presence of other substrates, and other organisms on the biodegradation of 2-chlorophenol (2-CP) was studied for Alcaligenes sp. isolated from natural sources. Assays of biodegradation were performed in batch and continuous-flow fluidized-bed aerobic reactors. Evaluation of biodegradation was performed by determining total phenols, chemical oxygen demand (COD), and 2-CP by ultraviolet (UV) spectrophotometry. Measurement of microbial growth was carried out by the plate count method. Bioassays of acute toxicity were performed to evaluate detoxification by using Daphnia magna. Results obtained show that under batch conditions with initial inocula of 106 cells/mL the strain grew exponentially with 100, 200, and 300 mg/L of 2-CP within 48 hr. A lag period was observed with low cell density inocula (105 cells/mL). The strain showed marked delay in the biodegradation of 2-CP at pH 5. Removal of target substrate from mixtures containing other carbon sources demonstrated the possibility of concurrent growth. Mineralization of 2-CP was assessed by gas chromatography carried out at the end of the batch assays and at the exit of the continuous-flow reactor. The presence of other organisms (bacteria, rotifers, ciliate, and algae) that developed in the fluidized-bed reactor did not affect the efficacy of the biodegradation of 2-CP. The removal of 2-CP in the two assayed systems was over 97% in all cases. Toxicity was not detected at the exit of the continuous reactor. © 2001 John Wiley & Sons, Inc. Environ Toxicol 16: 306,313, 2001 [source] Adding magnesium to the silver-gill binding model for rainbow trout (Oncorhynchus mykiss)ENVIRONMENTAL TOXICOLOGY & CHEMISTRY, Issue 3 2001Melissa L. Schwartz Abstract Rainbow trout (Oncorhynchus mykiss; 2,17 g) were exposed to approximately 0.1 ,M silver as AgNO3 for 3 to 4 h in synthetic, ion-poor water (20 ,M Ca, 100 ,M Na, 150 ,M Cl, pH 7) to which was added Mg, Ca, or thiosulfate (S2O3). Gills were extracted and assayed for Ag using graphite furnace atomic absorption spectrophotometry. Up to 210 mM Mg (four fold the concentration of Mg in seawater) did not reduce accumulation of Ag by trout gills. The conditional equilibrium stability constant (K) for Mg at silver-binding sites on the gills was calculated to be log KMg-gillAg = 3.0, or approximately half-as-strong binding as for Ca at these sites. The inclusion of the Mg-gill stability constant into the original Ag-gill binding model increases the flexibility of the model, although the competitive effects of Mg are only important in sodium-poor systems. [source] Influence of bacitracin on microbial functions in the gastrointestinal tract of horsesEQUINE VETERINARY JOURNAL, Issue 4 2000E. Collinder Summary This study investigated the influence of zinc bacitracin on the intestinal flora of horses. The functionally active intestinal flora was examined in 6 horses during treatment with zinc bacitracin. Utilising gas chromatography, spectrophotometry, gel electrophoresis and paper chromatography, samples were analysed on biochemical markers reflecting the action of parts of the intestinal flora. The following 5 flora-related functions were studied in faecal samples and intestinal samples from different sections of the hindgut: conversion of cholesterol to coprostanol and of bilirubin to urobilinogens, degradation of mucin and of ,-aspartylglycine and inactivation of tryptic activity. Conversion to coprostanol, conversion to urobilinogens and degradation of mucin were affected by treatment of zinc bacitracin and conversion to coprostanol was most sensitive. All functions were normalised in a short time, in contrast to man and rats. Differences in environmental exposures are probably the reason for a more rapid normalisation of the intestinal flora functions in horses. [source] Stability Constants and Dissociation Rates of the EDTMP Complexes of Samarium(III) and Yttrium(III)EUROPEAN JOURNAL OF INORGANIC CHEMISTRY, Issue 30 2008Ferenc Krisztián Kálmán Abstract The stability constants of Sm(EDTMP) (log,KML = 20.71) and Y(EDTMP) (log,KML = 19.19) were determined by a competition reaction between the Ln3+ ion (Ln3+ = Sm3+ or Y3+) and Cu2+ for the EDTMP ligand by spectrophotometry at pH , 10, in the presence of an excess amount of citrate (0.15 M NaCl, 25 °C). For determining the stability constants of Cu(EDTMP) (log,KML = 19.36) and Ca(EDTMP) (log,KML = 8.71) pH,potentiometry was used. In the pH range 4,9 the EDTMP complexes are present in the form of nonprotonated and mono-, di- and triprotonated species. The Ca2+ ion forms a dinuclear complex with Ln(EDTMP). In a simplified blood plasma model consisting of Sm3+, Ca2+ and Zn2+ metal ions, EDTMP, citrate, cysteine and histidine ligands, Sm3+ is practically present in the form of [Sm(HEDTMP)Ca]2,, whereas Zn2+ predominantly forms [Zn(HEDTMP)]5, and [Zn(H2EDTMP)]4, complexes. For studying the dissociation rates of the complexes, the kinetics of the metal exchange (transmetallation) reactions between the Ln(EDTMP) complexes and Cu2+,citrate were investigated in the pH range 7,9 by the stopped-flow method. The rates of the exchange reactions are independent of the Cu2+ concentration and increase with the H+ concentration. The rate constants, characterizing the proton-assisted dissociation of the Ln(EDTMP) complexes, are several orders of magnitude higher than those of the similar Ln(EDTA) complexes, because the protonation constants of Ln(EDTMP) are high and the protonated Ln(HEDTMP) and Ln(H2EDTMP) species are present in higher concentration. The half-times of dissociation of Sm(EDTMP) and Y(EDTMP) at pH = 7.4 and 25 °C are 4.9 and 7.5 s, respectively. These relatively short dissociation half-time values do not predict the deposition of Ln3+ ions in bones in the form of intact Ln(EDTMP) complexes. It is more probable that sorption of the EDTMP ligand and Sm3+ or Y3+ ions occurs independently after the dissociation of complexes.(© Wiley-VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2008) [source] Synthesis and Metal-Complexation Properties of a New Hydroxypyrimidinone-Functionalized SepharoseEUROPEAN JOURNAL OF INORGANIC CHEMISTRY, Issue 3 2005M. Alexandra Esteves Abstract The 1-hydroxy-2-(1H)-pyrimidinone derivative 4-(3-aminopropylamino)-1-hydroxy-2-(1H)-pyrimidinone (HOPY-PrN) was synthesized and its acid-base and complexation properties towards a set of metal ions (FeIII, AlIII, and ThIV) were studied by potentiometry and spectrophotometry. The ligand was further immobilized in an epoxy-activated sepharose by chemical coupling through the aminoalkyl pendent group with the aim of improving its sequestering capacity for residual amounts of metals. The new hydroxypyrimidinone-functionalized sepharose shows a high chelating capacity for metal ions in the pH range 3,8, thus suggesting good perspectives for potential environmental applications. (© Wiley-VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2005) [source] Lanthanide Chelates Based on Diethylenetriamine Fitted with O -Benzoic Acid Pendant ArmsEUROPEAN JOURNAL OF INORGANIC CHEMISTRY, Issue 7 2003Daniel Imbert Abstract A new polycarboxylate ligand H5L has been synthesized by the attachment of five benzoate subunits onto a diethylenetriamine framework. Seven pKa values have been determined by potentiometry, spectrophotometry and NMR spectroscopy as 1.9(2), 2.8(2), 3.87(5), 4.58(6), 4.87(6), 9.19(6) and 11.68(5), the first four corresponding to the carboxylic functions and the last three to amine sites. The interaction between H5L and LnIII ions in dilute aqueous solution has been examined by UV/Vis absorption and emission spectrometries, and has been found to result in monometallic complexes that are moderately stable in the pH range 3.7,7.5. Conditional stability constants at pH 5.3 are logK11 = 5.3(2), 6.6(1), 6.5(1) and 7.2(3) for La, Eu, Tb and Lu, respectively. In the case of TbIII, the stability constants for [Tb(HL)], and [Tb(H2L)] are log,111 = 22.0(2) and log,121 = 29.8(1), giving a pTb of 10.0. In the pH range 4,7, more than 90% of the TbIII ions are in the form of the neutral species [Tb(H2L)]. Lifetime determinations of the Eu(5D0) and Tb(5D4) excited levels in both H2O and D2O at pH 5.3 indicate 4.8 ± 0.5 (Eu) and 4.5 ± 0.5 (Tb) water molecules being bound in the inner coordination sphere of the LnIII. The triplet state of the ligand in water lies at around 26000 cm,1, resulting in a sizeable sensitisation of the Tb-centred luminescence (absolute quantum yield: ,abs = 10.3%), while the luminescence of EuIII is only poorly sensitised (,abs = 1.5%). (© Wiley-VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2003) [source] Oxidation of oleuropein studied by EPR and spectrophotometryEUROPEAN JOURNAL OF LIPID SCIENCE AND TECHNOLOGY, Issue 2 2008Evaggelia D. Tzika Abstract The autoxidation at alkaline pH and enzymatic oxidation by mushroom tyrosinase of oleuropein, the dominant biophenol present in the fruits and leaves of Olea europea, was followed by both electron paramagnetic resonance (EPR) and absorption spectroscopy. For comparison, the same oxidation processes were applied to 4-methylcatechol, a simple polyphenol present in olive mill wastewaters. EPR spectra of stable o -semiquinone radicals produced during autoxidation at pH,12 and short-lived o -semiquinone free radicals produced during autoxidation at pH,9.0 or tyrosinase action and stabilized by chelation with a diamagnetic metal ion (Mg2+) were recorded for both polyphenols, and the corresponding hyperfine splitting constants were determined. The UV-Vis spectral characteristics of the oxidation of polyphenols were highly dependent on the type of polyphenol, oxidant type and the pH of the reaction. The kinetic behavior of tyrosinase in the presence of oleuropein and 4-methylcatechol was followed by recording spectral changes at 400,nm (absorption maximum) over time. The tysosinase activity with oleuropein showed a pronounced pH optimum at pH,6.5 and a minor one around pH,8. From the data analysis of the initial rate at pH,6.5, the kinetic parameters Km = 0.34,±,0.03,mM and Vmax = 0.029,±,0.002 ,A400,min,1 were determined for oleuropein. [source] Nucleophilicities and Nucleofugalities of Organic Carbonates,EUROPEAN JOURNAL OF ORGANIC CHEMISTRY, Issue 22 2010Nicolas Streidl Abstract The kinetics of the reactions of the methyl carbonate ion with benzhydrylium ions in acetonitrile have been studied by UV/Vis spectrophotometry. Substitution of the resulting second-order rate constants and the electrophilicity parameters E of the benzhydrylium ions into the linear free energy relationship log,k = s(N + E) yielded the nucleophilicity parameters N25 = 16.03 and s25 = 0.64 for methyl carbonate in acetonitrile. The kinetics of the reverse reactions, i.e., of the solvolyses of ring-substituted benzhydryl alkyl carbonates in different aqueous solvents were followed by conductimetry. The obtained first-order rate constants and the known electrofugality parameters Ef of benzhydrylium ions were used to determine the nucleofugality parameters Nf and sf of the ROCO2, groups by using the linear free energy relationship log,k = sf(Nf + Ef). The leaving group abilities of carbonates decrease by a factor of about 300 from PhOCO2, over MeOCO2, and iBuOCO2, to tBuOCO2, in various alcoholic and aqueous solvents. tert -Butyl carbonates (tBocO-R) are, thus, considerably more stable with respect to heterolytic cleavage of the O,R bond than other organic carbonates. [source] Magnesium sulphate treatment decreases blood,brain barrier permeability during acute hypertension in pregnant ratsEXPERIMENTAL PHYSIOLOGY, Issue 2 2008Anna G. Euser Eclampsia is associated with increased blood,brain barrier (BBB) permeability and formation of cerebral oedema. Magnesium sulphate is used to treat eclampsia despite an unclear mechanism of action. This study was to determine the effect of magnesium sulphate on in vivo BBB permeability and formation of cerebral oedema during acute hypertension and on brain aquaporin-4 (AQP4) protein expression. An in vivo model of hypertensive encephalopathy was used in late-pregnant (LP) rats following magnesium sulphate treatment, 270 mg kg,1i.p. injection every 4 h for 24 h. Permeability of the BBB was determined by in situ brain perfusion of Evan's Blue (EB) and sodium fluorescein (NaFl), and dye clearance determined by fluorescence spectrophotometry. Cerebral oedema was determined following acute hypertension by measuring brain water content. The effect of magnesium treatment on AQP4 expression was determined by Western blot analysis. Acute hypertension with autoregulatory breakthrough increased BBB permeability to EB in both brain regions studied (P < 0.05). Magnesium attenuated BBB permeability to EB during acute hypertension by 41% in the posterior cerebrum (P < 0.05) but had no effect in the anterior cerebrum (P > 0.05). Treatment with magnesium did not change NaFl permeability, cerebral oedema formation or AQP4 expression. In summary, BBB permeability to Evan's Blue was increased by acute hypertension in LP rats, and this was attenuated by treatment with magnesium sulphate. The greatest effect on BBB permeability to EB was in the posterior cerebrum, an area particularly susceptible to oedema formation during eclampsia. [source] Molecular responses of Campylobacter jejuni to cadmium stressFEBS JOURNAL, Issue 20 2008Nadeem O. Kaakoush Cadmium ions are a potent carcinogen in animals, and cadmium is a toxic metal of significant environmental importance for humans. Response curves were used to investigate the effects of cadmium chloride on the growth of Camplyobacter jejuni. In vitro, the bacterium showed reduced growth in the presence of 0.1 mm cadmium chloride, and the metal ions were lethal at 1 mm concentration. Two-dimensional gel electrophoresis combined with tandem mass spectrometry analysis enabled identification of 67 proteins differentially expressed in cells grown without and with 0.1 mm cadmium chloride. Cellular processes and pathways regulated under cadmium stress included fatty acid biosynthesis, protein biosynthesis, chemotaxis and mobility, the tricarboxylic acid cycle, protein modification, redox processes and the heat-shock response. Disulfide reductases and their substrates play many roles in cellular processes, including protection against reactive oxygen species and detoxification of xenobiotics, such as cadmium. The effects of cadmium on thioredoxin reductase and disulfide reductases using glutathione as a substrate were studied in bacterial lysates by spectrophotometry and nuclear magnetic resonance spectroscopy, respectively. The presence of 0.1 mm cadmium ions modulated the activities of both enzymes. The interactions of cadmium ions with oxidized glutathione and reduced glutathione were investigated using nuclear magnetic resonance spectroscopy. The data suggested that, unlike other organisms, C. jejuni downregulates thioredoxin reductase and upregulates other disulfide reductases involved in metal detoxification in the presence of cadmium. [source] A novel extracellular esterase from Bacillus subtilis and its conversion to a monoacylglycerol hydrolaseFEBS JOURNAL, Issue 21 2000Thorsten Eggert A novel gene lipB, which encodes an extracellular lipolytic enzyme, was identified in the Bacillus subtilis genomic DNA sequence. We have cloned and overexpressed lipB in B. subtilis and Escherichia coli and have also purified the enzyme from a B. subtilis culture supernatant to electrophoretic homogeneity. Four different lipase assays were used to determine its catalytic activity: pH-stat, spectrophotometry, fluorimetry and the monomolecular film technique. LipB preferentially hydrolysed triacylglycerol-esters and p -nitrophenyl-esters of fatty acids with short chain lengths of ,,10 carbon atoms. Triolein, which is a typical substrate for true lipases, was not hydrolysed at all. These results led us to classify LipB as an esterase rather than a lipase. The catalytic triad of LipB consists of residues Ser78, Asp134, and His157 as demonstrated by amino-acid sequence alignments and site-directed mutagenesis. The nucleophile Ser78 is located in a lipase-specific consensus sequence, which is Ala-X-Ser-X-Gly for most Bacillus lipases. All other bacterial lipases contain a glycine residue instead of the alanine at position-2 with respect to the catalytic serine. We have investigated the role of this alanine residue by constructing LipB variant A76G, thereby restoring the lipase-specific consensus motif. When compared with LipB this variant showed a markedly reduced thermostability but an increased stability at pH 5,7. Determination of the specific activities of wild-type LipB and variant A76G using a monomolecular film of the substrate monoolein revealed an interesting result: the A76G substitution had converted the esterase LipB into a monoacylglycerol hydrolase. [source] Improvements for comparative analysis of changes in diversity of microbial communities using internal standards in PCR-DGGEFEMS MICROBIOLOGY ECOLOGY, Issue 3 2005Dorthe Groth Petersen Abstract The use of internal standards both during DNA extraction and PCR-DGGE procedure gives the opportunity to analyse the relative abundance of individual species back to the original sample, thereby facilitating relative comparative analysis of diversity. Internal standards were used throughout the DNA extraction and PCR-DGGE to compensate for experimental variability. Such variability causes decreased reproducibility among replicate samples as well as compromise comparisons between samples, since experimental errors cannot be differentiated from actual changes in the community abundance and structure. The use of internal standards during DNA extraction and PCR-DGGE is suitable for ecological and ecotoxicological experiments with microbial communities, where relative changes in the community abundance and structure are studied. We have developed a protocol Internal Standards in Molecular Analysis of Diversity (ISMAD) that is simple to use, inexpensive, rapid to perform and it does not require additional samples to be processed. The internal standard for DNA extraction (ExtrIS) is a fluorescent 510-basepair PCR product which is added to the samples prior to DNA extraction, recovered together with the extracted DNA from the samples and analysed with fluorescence spectrophotometry. The use of ExtrIS during isolation of sample DNA significantly reduced variation among replicate samples. The PCR internal standard (PCRIS) originates from the Drosophila melanogaster genome and is a 140-basepair long PCR product, which is amplified by non-competitive primers in the same PCR reaction tubes as the target DNA and analysed together with the target PCR product on the same DGGE gel. The use of PCRIS during PCR significantly reduced variation among replicate samples both when assessing total PCR product and when comparing bands representing species on a DGGE gel. The entire ISMAD protocol was shown to accurately describe changes in relative abundance in an environmental sample using PCR-DGGE. It should, however, be mentioned that despite the use of ISMAD some inherent biases still exist in DNA extraction and PCR-DGGE and these should be taken into consideration when interpreting the diversity in a sample based on a DGGE gel. [source] Study of the mechanism of microwave-assisted extraction of Mahonia bealei (Fort.) leaves and Chrysanthemum morifolium (Ramat.) petalsFLAVOUR AND FRAGRANCE JOURNAL, Issue 3 2004Shan Gao Abstract A study of microwave-assisted extraction (MAE) for berberine in Mahonia bealei (Fort.) was carried out with batch equipment, in order to investigate the mechanism of the extraction related to structural changes in the glands. The extracts were analysed by ultraviolet-visible spectrophotometry at 347 nm. The parameters investigated were solvent types, the intensity of microwave energy and the process ratio (g/ml) of materials to solvent volume. The microwave-assisted extraction of different moisture content of materials was developed and optimized by means of three-factor and three-level orthogonal designs. Electron and optical micrographs of M. bealei (Fort.) leaves and Chrysanthemum morifolium (Ramat.) petals showed that the mechanism of the extractions was related to structural changes in the plant cells. Copyright © 2004 John Wiley & Sons, Ltd. [source] Analysis of adobe wall composition at the Chaves-Hummingbird Site, New Mexico, by diffuse reflectance spectrophotometryGEOARCHAEOLOGY: AN INTERNATIONAL JOURNAL, Issue 8 2007William Balsam This article investigates adobe wall construction materials utilized by prehistoric inhabitants of Chaves-Hummingbird Pueblo, an ancestral Pueblo village located ,20 miles west of Albuquerque, New Mexico. The walls were constructed with native clay-rich soils some time between approximately 1275,1450 A.D. Samples were analyzed with a diffuse reflectance spectrophotometer from the near ultraviolet (NUV) through the visible (VIS) and into the near infrared (NIR) region of the electromagnetic spectrum. Cluster analysis of samples from 275 adobe walls and 36 soil locations surrounding the pueblo room blocks indicates four clusters. Comparison of typical samples from the four clusters indicates that they are very similar and are distinguished by minor variations in the three primary spectrally determined components, Na-Ca montmorillonite, bentonite, and goethite. In general, clusters correspond with room construction episodes that are discernible through patterns of wall bonding and abutment recorded during the archaeological investigation of the site. This suggests that during different phases of construction the source of the wall adobe changed. Many of the soil samples are included in wall clusters and therefore reveal a potential source of material used for adobe, adjacent soils. However, not all the soil surrounding the pueblo grouped with wall clusters indicating a preference for certain soil types and that some soils were probably unsuitable for making adobe. Therefore, diversity in spectrally identified construction materials provides insights into source locations and possible construction preferences of the site inhabitants. © 2007 Wiley Periodicals, Inc. [source] Preparation and Characteristics of Esculin-Imprinted PolymersHELVETICA CHIMICA ACTA, Issue 6 2007Guo-Song Wang Abstract Four molecularly imprinted polymers (MIPs) were prepared in MeOH with esculin (=6,7-dihydroxycoumarin 6-(, - D -glucopyranoside)=6-(, - D -glucopyranosyloxy)-7-hydroxy-2H -1-benzopyran-2-one) as the imprinted molecule, methacrylic acid (=2-methylprop-2-enoic acid; MAA), acrylamide (=prop-2-enamide; AM), 4-vinylpyridine (=4-ethenylpyridine; 4-VP), or 2-vinylpyridine (=2-ethenylpyridine; 2-VP) as the functional monomer, respectively, as well as ethylene glycol dimethacrylate (=2-methylprop-2-enoic acid ethane-1,2-diyl ester; EGDMA) as the cross-linking agent. The interaction between the template and the functional monomers was investigated by fluorescence and UV spectrophotometry, respectively, which revealed the presence of esculin/monomer complexes in the stoichiometric ratio 1,:,2 in the pre-polymerization mixture. The resultant polymers were studied in equilibrium binding experiments to evaluate the recognition ability and the binding capacity towards esculin. The results showed that MIP1, prepared with MAA as the functional monomer, exhibited advantageous characteristics of high binding capacity, optimal imprinting effect, and good selectivity towards esculin. The Scatchard analysis indicated that there are two types of binding sites in MIP1, and its binding parameters including the apparent maximum numbers of binding sites and the dissociation constants were calculated. Finally, by packing an SPE column (SPE=solid-phase extraction) with MIP1, the esculin was separated and enriched successfully by this sorbent from samples of Cortex fraxini, and the average recovery was up to 74.7%. [source] Photoinduced Formation of Reactive Oxygen Species from the Acid Form of 6-(Hydroxymethyl)pterin in Aqueous SolutionHELVETICA CHIMICA ACTA, Issue 6 2006Andrés Abstract The photochemistry of 6-(hydroxymethyl)pterin (HPT; 1) in aqueous solution (pH 5,6) was investigated by irradiation at 350,nm at room temperature. The photochemical reactions of the acidic form 1a were followed by UV/VIS spectrophotometry, thin-layer chromatography (TLC), high-performance liquid chromatography (HPLC), and enzymatic methods for the determination of the superoxide anion radical (O) and hydrogen peroxide (H2O2). When 1a is exposed to UV-A radiation, the intermediates 4 and 4, are formed reacting with O2 to yield 6-formylpterin (FPT; 5) and 6-carboxypterin (CPT; 6) under formation of O and H2O2 (Scheme,3). The quantum yields of the disappearance of HPT (1a) and of the formation of the photoproducts 5 and 6 were determined. HPT was investigated for its efficiency in singlet-oxygen (1O2) production in acidic aqueous solution. The corresponding quantum yield of 1O2 production (,,) was 0.15,±,0.02, as measured by the 1O2 luminescence in the near-IR (1270,nm) upon continuous excitation of the sensitizer. However, 1O2 does not participate in the actual photooxidation of HPT (1a) to FPT (5) and CPT (6). [source] Paraoxonase-1 (PON1) activity as a risk factor for atherosclerosis in chronic renal failure patientsHEMODIALYSIS INTERNATIONAL, Issue 4 2008Saeed Abdelwhab SAEED Abstract Paraoxonase is a high-density lipoprotein-associated enzyme and has been shown to reduce the susceptibility to low-density lipoprotein peroxidation. This study aimed to investigate the activity of serum paraoxonase in uremic patients on hemodialysis (HD) and in the predialysis period, and to evaluate the correlations of vascular disease with paraoxonase activity. Thirty patients with chronic renal failure (CRF) undergoing HD (group 1), 30 patients with CRF under conservative treatment (group 2), and 30 healthy controls (group 3) were included. Basal, salt-stimulated, and arylesterase activity were tested by UV spectrophotometry. Serum lipid parameters were determined. B-Mode Doppler ultrasound was used to assess common carotid intima-media thickness (IMT). Basal paraoxonase, salt-stimulated, and arylesterase activity showed no significant difference between group 1 and group 2. However, it was significantly lower in group 1 and in group 2 than controls. Carotid IMT was significantly higher in group 1 than group 2 and both were significantly higher than controls. Basal paraoxonase-1 (PON1), salt-stimulated PON1, and arylesterase activity correlate with BUN, but only basal PON1 and salt-stimulated PON1 correlate with serum albumin. Linear regression showed that the most significant determinant of carotid IMT was PON1 arylesterase activity in group 1 and arylesterase activity and basal PON1 activity in group 2. Patients with CRF, whether under HD or conservative treatment, have reduced basal and stimulated paraoxonase activities, and this could be an important factor causing increased vascular disease in those patients. Modifying this factor can be of great value to protect against this common complication. [source] Liver damage underlying unexplained transaminase elevation in human immunodeficiency virus-1 mono-infected patients on antiretroviral therapy,HEPATOLOGY, Issue 2 2009Patrick Ingiliz Liver damage associated with chronic unexplained high serum transaminases in human immunodeficiency virus (HIV)-infected patients under combined antiretroviral therapy is unknown. Liver histology was prospectively investigated in patients presenting serum transaminase elevation for more than 6 months, after exclusion of alcohol abuse, hepatitis C virus (HCV) or hepatitis B virus (HBV) infection, autoimmune, and genetic liver diseases. In a subgroup of patients, liver mitochondrial activities were measured by spectrophotometry and mitochondrial DNA (mtDNA) by real-time polymerase chain reaction (PCR). Thirty patients were included with median values of alanine aminotransferase (ALT) levels: 80 U/L, age: 46 years, body mass index: 23 kg/m2, HIV RNA: 200 copies/mL, CD4 count: 365/mm3, duration of HIV infection: 13 years, and duration of treatment exposure: 118, 41, and 53 months for nucleoside reverse transcriptase inhibitors, non-nucleoside reverse transcriptase inhibitors, and protease inhibitors, respectively. Histological anomalies were found in 22 of 30 patients. Steatosis was present in 18 patients, severe in nine patients, and associated with inflammation in 16 patients with a diagnosis of non-alcoholic steatohepatitis (NASH). Fibrosis was found in 18 patients, severe in six patients and associated with steatosis in 13 patients. Significant liver respiratory complex I defect, contrasting with high complex IV activity and normal mitochondrial DNA content, was observed in the group of patients compared with controls. The presence of NASH was correlated with high fasting glycemia and insulin levels, not with liver mitochondrial function or mitochondrial DNA content. Conclusions: HIV-infected patients on combined antiretroviral therapy with chronic transaminase elevation of unknown origin have a high rate of liver lesions, mostly consistent with NASH related to insulin resistance. (HEPATOLOGY 2008.) [source] Decalcification of root canal dentine by citric acid, EDTA and sodium citrateINTERNATIONAL ENDODONTIC JOURNAL, Issue 6 2004L. F. Machado-Silveiro Abstract Aim, To measure the demineralization capability of 1 and 10% citric acid, 10% sodium citrate and 17% EDTA during immersions of 5, 10 and 15 min on root canal dentine. Methodology, Crowns were sectioned from eight maxillary canines. The cementum was removed from the cervical third of the roots to expose the dentine. Canals were prepared using a handpiece-mounted Largo Peeso reamer. A 3-mm thick cross-sectional slice was obtained from the cervical third of each root. Each slice was sectioned into four equal parts. These specimens were assigned to one of four groups (n = 8) for the application of 1% citric acid, 10% citric acid, 10% sodium citrate or 17% EDTA. Each specimen underwent three successive 5-min immersions in each solution at room temperature. The solutions were not renewed between immersions. Two millimetres of solution were collected from the extracts and lanthanum oxide was added for the calcium reading by spectrophotometry. To compare the amounts of calcium removed by each solution, the Friedman test was used for the global comparison and the Wilcoxon test for paired comparisons. Differences between groups were evaluated using the Kruskal,Wallis test for the global comparison and Mann,Whitney test for paired comparisons. Results, Overall, 1 and 10% citric acid were more effective than EDTA or sodium citrate at the three immersion times (P < 0.001); 10% citric acid was more effective than 1% citric acid (P < 0.001). EDTA and 1 and 10% citric acid showed decreasing effectiveness with time, and the decrease was significant for citric acid at both concentrations (P < 0.001). Although sodium citrate removed little calcium during the three time periods, the small increase recorded was significant (P < 0.01). Conclusions, Citric acid at 10% was the most effective decalcifying agent, followed by 1% citric acid, 17% EDTA and 10% sodium citrate. [source] | |||||||||||||||||||||||||||||||||||||||||||