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High-pressure Liquid Chromatography (high-pressure + liquid_chromatography)
Selected AbstractsLeaf Disc Inoculation, a Fast and Precise Test for the Screening of Metalaxyl Tolerance in Sunflower Downy MildewJOURNAL OF PHYTOPATHOLOGY, Issue 6 2001B. Rozynek A new method, based on leaf disc inoculation, was developed for the screening of metalaxyl tolerance in field isolates of Plasmopara halstedii. High-pressure liquid chromatography (HPLC) was used to determine the fungicide concentration in the inoculation medium and in the incubated leaf tissue over the test period. These measurements revealed that the fungicide concentration inside the leaf tissue within 24 h had adjusted to the concentration in the outer medium and remained constant for the time of cultivation over a period of more than 11 days. In contrast to whole seedling tests with application of the fungicide via seed dressing, the leaf disc method allows precise quantification of the effective fungicide concentration at the site of infection and is less space and time consuming. Metalaxyl tolerance of P. halstedii isolates was gradually determined according to the sporulation of the pathogen on sunflower leaf discs in the presence of increasing fungicide concentrations. Isolates collected in South Germany showed no tolerance and sporulation was prohibited when tests were carried out at 0.02 ,g (a.i.)/ml of metalaxyl or more. In contrast, a tolerant French isolate developed sporangia on leaf discs incubated in a metalaxyl solution of 100 ,g (a.i.)/ml. [source] Inhibition of inducible nitric oxide synthase reduces lipopolysaccharide-induced renal injury in the ratCLINICAL AND EXPERIMENTAL PHARMACOLOGY AND PHYSIOLOGY, Issue 12 2004M Kadkhodaee SUMMARY 1.,Gram-negative bacterial lipopolysaccharide (LPS) release and subsequent septic shock is a major cause of death in intensive care units. Lipopolysaccharide has been reported to increase the production of nitric oxide (NO) and the formation of oxygen-derived free radicals (OFR) in different organs. The aim of the present study was to evaluate the role of an inducible form of NO synthase (iNOS) and OFR production in LPS-induced renal impairment. 2.,Measurement of vitamin E as the most important fat-soluble anti-oxidant was used as a marker of tissue oxidative stress. Lipopolysaccharide (10 mg/kg), l -iminoethyl lysine (L-Nil; 3 mg/kg, i.p.; a specific inhibitor of iNOS activity) and dimethyl thiourea (DMTU; 500 mg/kg i.p.; a well-known OFR scavenger) were used. Four groups of eight rats were studied. One group received LPS, whereas a second group received LPS + L-Nil. A third group received LPS + DMTU and the fourth group, receiving saline, acted as a control group. To evaluate renal function, plasma creatinine and blood urea nitrogen (BUN) were measured. High-pressure liquid chromatography and ultraviolet detection were used to measure plasma and tissue vitamin E levels. Light microscopy was used to examine histopathological changes in the four groups. 3.,Lipopolysaccharide markedly decreased the vitamin E content of renal plasma and tissue (P < 0.05). Administration of L-Nil attenuated renal dysfunction and preserved vitamin E levels. However, DMTU failed to prevent renal injury, as indicated by plasma BUN levels and renal histology, despite the fact that it maintained renal vitamin E levels and increased plasma vitamin E levels. Thus, the overproduction of NO by iNOS may have a role in this model of LPS-induced renal impairment. [source] Altered Tryptophan Metabolism in the Brain of Cystatin B -Deficient Mice: A Model System for Progressive Myoclonus EpilepsyEPILEPSIA, Issue 10 2006Annika Vaarmann Summary:,Purpose: Progressive myoclonus epilepsy of the Unverricht,Lundborg type (EPM1) is a rare neurologic disorder, associated with mutations in the Cystatin B (Cstb) gene. Mice lacking Cstb, a cysteine protease inhibitor of the cathepsine family of proteases, provide a mammalian model for EPM1 by displaying similarly progressive ataxia, myoclonic seizures, and neurodegeneration. However, the linkage of Cstb deficit on the molecular level to pathologic features like myoclonic jerks or tonic,clonic seizures has remained unclear. We examined the tryptophan (TRP) metabolism, along the serotonin (5HT) and kynurenine (KYN) pathway in the brain of Cstb -deficient mice, in relation to their possible involvement in the seizure phenotype. Methods: TRP and its metabolites, along the 5HT and KYN pathways, were assayed in brain tissue by high-pressure liquid chromatography (HPLC) with electrochemical detection. The inverted wire grid and mild handling tests were used for evaluation of ataxia and myoclonic activity. Results: The Cstb -deficient mice had constitutively increased TRP, 5HT, and 5-hydroxyindole acetic acid (5HIAA) levels in the cerebral cortex and cerebellum and increased levels of KYN in the cerebellum. These neurochemical changes were accompanied with ataxia and an apparent myoclonic phenotype among the Cstb -deficient mice. Conclusions: Our findings suggest that secondary processes (i.e., overstimulation of serotoninergic transmission) on the cellular level, initiated by Cstb deficiency in specific brain regions, may be responsible for the myoclonic/seizure phenotype in EPM1. [source] Plasma Concentrations of Risperidone and Olanzapine during Coadministration with OxcarbazepineEPILEPSIA, Issue 5 2005Maria Rosaria Muscatello Summary:,Purpose: Oxcarbazepine (OZC) is a second-generation antiepileptic drug (AED) that also may be used as a mood stabilizer. Unlike carbamazepine (CBZ), which is an inducer of the cytochrome P-450 isoforms and may accelerate the elimination of several therapeutic agents, OXC seems to have only a modest inducing action. The aim of this investigation was to evaluate the effect of a treatment with OXC on plasma concentrations of the new antipsychotics risperidone and olanzapine. Methods: OXC, at a dosage of 900,1,200 mg/day, was administered for 5 consecutive weeks to 25 outpatients, 10 men and 15 women, aged 25 to 64 years, with bipolar or schizoaffective disorder. Twelve patients were stabilized on risperidone therapy (2,6 mg/day) and 13 on olanzapine (5,20 mg/day). Steady-state plasma concentrations of risperidone and its active metabolite 9-hydroxyrisperidone (9-OH-risperidone) and olanzapine were measured by high-pressure liquid chromatography (HPLC) before addition of OXC and after 5 weeks from the start of adjunctive treatment. Results: OXC caused only minimal and no significant changes in the mean plasma levels of risperidone (from 5.6 ± 3.6 ng/ml at baseline to 4.8 ± 2.6 ng/ml at week 5), 9-OH-risperidone (from 23.6 ± 7.5 to 24.7 ± 7.4 ng/ml), and olanzapine (from 26.5 ± 5.7 ng/ml at baseline to 27.8 ± 5.1 ng/ml). OXC coadministration with either risperidone or olanzapine was well tolerated. Conclusions: Our findings indicate that OXC does not affect the elimination of risperidone and olanzapine, thus confirming its weak inducing effect on hepatic drug-metabolizing enzymes. [source] Ret deficiency in mice impairs the development of A5 and A6 neurons and the functional maturation of the respiratory rhythmEUROPEAN JOURNAL OF NEUROSCIENCE, Issue 10 2005J. C. Viemari Abstract Although a normal respiratory rhythm is vital at birth, little is known about the genetic factors controlling the prenatal maturation of the respiratory network in mammals. In Phox2a mutant mice, which do not express A6 neurons, we previously hypothesized that the release of endogenous norepinephrine by A6 neurons is required for a normal respiratory rhythm to occur at birth. Here we investigated the role of the Ret gene, which encodes a transmembrane tyrosine kinase receptor, in the maturation of norepinephrine and respiratory systems. As Ret -null mutants (Ret,/,) did not survive after birth, our experiments were performed in wild-type (wt) and Ret,/, fetuses exteriorized from pregnant heterozygous mice at gestational day 18. First, in wt fetuses, quantitative in situ hybridization revealed high levels of Ret transcripts in the pontine A5 and A6 areas. Second, in Ret,/, fetuses, high-pressure liquid chromatography showed significantly reduced norepinephrine contents in the pons but not the medulla. Third, tyrosine hydroxylase immunocytochemistry revealed a significantly reduced number of pontine A5 and A6 neurons but not medullary norepinephrine neurons in Ret,/, fetuses. Finally, electrophysiological and pharmacological experiments performed on brainstem ,en bloc' preparations demonstrated impaired resting respiratory activity and abnormal responses to central hypoxia and norepinephrine application in Ret,/, fetuses. To conclude, our results show that Ret gene contributes to the prenatal maturation of A6 and A5 neurons and respiratory system. They support the hypothesis that the normal maturation of the respiratory network requires afferent activity corresponding to the A6 excitatory and A5 inhibitory input balance. [source] In vitro effects of tacrolimus on human cytochrome P450FUNDAMENTAL & CLINICAL PHARMACOLOGY, Issue 6 2002K. Lecointre Abstract Tacrolimus, a potent immunosuppressive drug, is known to be metabolized predominantly in the liver by cytochrome P450 3A (CYP3A). In order to determine the potential of tacrolimus to inhibit the metabolism of other drugs, we have investigated its inhibitory effects on specific cytochrome reactions. Specific substrates for the seven cytochromes (CYPs) 1A2, 2A6, 2C9, 2C19, 2D6, 2E1 and 3A4/5 were incubated with human hepatic microsome preparations with or without specific inhibitors or tacrolimus and the metabolites were detected by high-pressure liquid chromatography (HPLC) or fluorimetric methods. All the specific inhibitors reduced or abolished the specific CYP activity. Tacrolimus had no effect on any CYP at concentrations below 1 µm, while at higher concentrations it had a mild inhibitory effect on CYP3A4 and 3A5. These observations suggest that tacrolimus is unlikely to potentiate the effect of coadministered drugs through inhibition of their metabolism in the liver. [source] The Influence of Lactobacillus brevis on Ornithine Decarboxylase Activity and Polyamine Profiles in Helicobacter pylori -Infected Gastric MucosaHELICOBACTER, Issue 2 2004Michele Linsalata ABSTRACT Background., Functional probiotics may prevent Helicobacter pylori infection, and some evidence suggests that they also possess antitumor properties. Lactobacillus brevis (CD2) is a functional Lactobacillus strain with peculiar biochemical features, essentially related to the activity of arginine deiminase. This enzyme catalyzes the catabolism of arginine and affects the biosynthesis of polyamines (putrescine, spermidine, and spermine). Polyamines are polycations found in high concentrations in both normal and neoplastic cells. Our aims were: 1, to assess whether oral administration of L. brevis (CD2) affects H. pylori survival in the human gastric mucosa; 2, to evaluate the effects of L. brevis (CD2) on polyamine biosynthesis in gastric biopsies from H. pylori- positive patients. Materials and Methods., For 3 weeks before endoscopy, 22 H. pylori- positive dyspeptic patients randomly received (ratio 1 : 1) high oral doses of L. brevis (CD2) or placebo. Before and after treatment, H. pylori infection was determined by urea breath test (UBT). In gastric biopsies, ornithine decarboxylase activity and polyamine levels were, respectively, evaluated by a radiometric technique and high-pressure liquid chromatography (HPLC). Results.,L. brevis (CD2) treatment did not eradicate H. pylori. However, a reduction in the UBT delta values occurred, suggesting a decrease in intragastric bacterial load. Significantly, L. brevis (CD2) induced a decrease in gastric ornithine decarboxylase activity and polyamine levels. Conclusions., Our data support the hypothesis that L. brevis (CD2) treatment decreases H. pylori colonization, thus reducing polyamine biosynthesis. Alternatively, the arginine deiminase activity following L. brevis (CD2) administration might cause arginine deficiency, preventing polyamine generation from gastric cells. [source] Oxidative damage is increased in human liver tissue adjacent to hepatocellular carcinomaHEPATOLOGY, Issue 6 2004Christoph Jüngst Accumulation of genetic alterations in hepatocarcinogenesis is closely associated with chronic inflammatory liver disease. 8-oxo-2,-deoxyguanosine (8-oxo-dG), the major promutagenic DNA adduct caused by reactive oxygen species (ROS), leads to G:C , T:A transversions. These lesions can be enzymatically repaired mainly by human MutT homolog 1 (hMTH1), human 8-oxo-guanine DNA glycosylase (hOGG1) and human MutY homolog (hMYH). The aim of this study was to evaluate the extent of oxidative damage and its dependence on the cellular antioxidative capacity and the expression of specific DNA repair enzymes in tumor (tu) and corresponding adjacent nontumor (ntu) liver tissue of 23 patients with histologically confirmed hepatocellular carcinoma. 8-oxo-dG levels, as detected by high-pressure liquid chromatography with electrochemical detection, were significantly (P = .003) elevated in ntu tissue (median, 129 fmol/,g DNA) as compared to tu tissue (median, 52 fmol/,g DNA), and were closely associated with inflammatory infiltration. In ntu tissue, the hepatic iron concentration and malondialdehyde levels were significantly (P = .001) higher as compared to tu tissue. Glutathione content, glutathione peroxidase activity and manganese superoxide dismutase messenger RNA (mRNA) expression did not show statistical differences between ntu and tu tissue. Real-time reverse transcription polymerase chain reaction revealed in tu tissue significantly (P = .014) higher hMTH1 mRNA expression compared to ntu tissue. In contrast, hMYH mRNA expression was significantly (P < .05) higher in ntu tissue. No difference in hOGG1 mRNA expression was seen between tu and ntu. In conclusion, these data suggest that ROS generated by chronic inflammation contribute to human hepatocarcinogenesis. The role of DNA repair enzymes appears to be of reactive rather than causative manner. (HEPATOLOGY 2004;39:1663,1672.) [source] Naturally occurring polyphenolic antioxidants modulate IgE-mediated mast cell activationIMMUNOLOGY, Issue 4 2000S.-S. Chen Summary Reactive oxygen species (ROS) are known to modulate activities of a host of kinases, phosphatases and transcription factors. Rutin and chlorogenic acid (CGA) are the major polyphenolic antioxidants present in the small molecular fraction of smokeless tobacco leaf extracts, as ascertained by reverse-phase high-pressure liquid chromatography (HPLC) and mass spectrometry. Levels of intracellular ROS in resting versus antigen,immunoglobulin E (IgE)-challenged murine mast cells were measured at 510 nm by fluorescence-activated cell sorting (FACS) using carboxy-dichlorofluorescein (DCFH-DA). Enhanced ROS production was observed in IgE-sensitized mast cells following antigenic challenge. Rutin and CGA reduced ROS levels in antigen,IgE-activated mast cells. Concomitantly, they also profoundly inhibited histamine release by these activated mast cells. In contrast, rutin and CGA augmented the inducible cytokine messages, i.e. interleukin (IL)-10, IL-13, interferon-, (IFN-,), IL-6 and tumour necrosis factor-, (TNF-,) in IgE-sensitized mast cells following antigen challenge. This study indicates that tobacco polyphenolic antioxidants that quench intracellular ROS, differentially affect two effector functions of antigen,IgE-activated mast cells. This model system may be employed to determine the molecular target of polyphenols. The potential role of these polyphenolic antioxidants on IgE-mediated allergy in vivo depends on a balance of their differential effects on mast cell activation. [source] Prevalence of BRCA1 and BRCA2 mutations in Pakistani breast and ovarian cancer patientsINTERNATIONAL JOURNAL OF CANCER, Issue 12 2006Muhammad U. Rashid Abstract Among Asian countries, Pakistan has the highest rates of breast and ovarian cancer. To assess the contribution of the BRCA1 and BRCA2 germ line mutations to these high rates, we conducted the first study of 176 Pakistani breast and ovarian cancer patients, selected on family history and on age of diagnosis. Comprehensive BRCA mutation screening was performed using a range of techniques, including denaturing high-pressure liquid chromatography, single strand conformational polymorphism analysis and protein truncation test, followed by DNA sequencing. Thirty deleterious germ-line mutations were identified in the 176 families (17.0%), including 23 in BRCA1 and 7 in BRCA2. Four mutations, 185delAG, 185insA, S1503X and R1835X, were recurrent; these accounted for 52% of all identified BRCA1 mutations. Haplotype analyses suggested founder effects for 3 of these. The prevalence of BRCA1 or BRCA2 mutations was 42.8% for families with multiple cases of breast cancer, and was 50.0% for the breast/ovarian cancer families. The prevalence of mutations was 11.9% for single cases of early-onset breast cancer (,30 years) and was 9.0% for single cases of early-onset ovarian cancer (,45 years). Our findings show that BRCA mutations account for a substantial proportion of hereditary breast/ovarian cancer and early-onset breast and ovarian cancer cases in Pakistan. © 2006 Wiley-Liss, Inc. [source] Explicitly correlated SCF study of anharmonic vibrations in (H2O)2INTERNATIONAL JOURNAL OF QUANTUM CHEMISTRY, Issue 4-5 2002Donald D. Shillady Abstract Modeling solvation in high-pressure liquid chromatography (HPLC) requires calculation of anharmonic vibrational frequencies of solvent clusters for a statistical partition function. An efficient computational method that includes electron correlation is highly desirable for large clusters. A modified version of the "soft Coulomb hole" method of Chakravorty and Clementi has recently been implemented in a Gaussian-lobe-orbital (GLO) program (PCLOBE) to include explicit electron,electron correlation in molecules. The soft Coulomb hole is based on a modified form of Coulomb's law: An algorithm has been developed to obtain the parameter "w" from a polynomial in the effective scaling of each primitive Gaussian orbital relative to the best single Gaussian of the H1s orbital. This method yields over 90% of the correlation energy for molecules of low symmetry for which the original formula of Chakravorty and Clementi does not apply. In this work, all the vibrations of the water dimer are treated anharmonically. A quartic perturbation of the harmonic vibrational modes is constrained to be equal to the exact Morse potential eigenvalue based on a three-point fit. This work evaluates the usefulness of fitting a Morse potential to a hydrogen bond vibrational mode and finds it to be slightly better than using MP2 vibrational analysis for this important dimer. A three-point estimate of the depth, De, of a Morse potential leads to a correction formula for anharmonicity in terms of the perturbed harmonic frequency: When scaled by 0.9141, the harmonic Morse method leads to essentially the same results as scaling the BPW91 local density method by 0.9827. © 2002 Wiley Periodicals, Inc. Int J Quantum Chem, 2002 [source] Preparation and in vitro release of D,L -tetrahydropalmatine-loaded graft copolymer nanoparticlesJOURNAL OF APPLIED POLYMER SCIENCE, Issue 6 2008Yinglei Zhai Abstract D,L -tetrahydropalmatine (THP)-loaded poly{[,-maleic anhydride-,-methoxy-poly(ethylene glycol)]- co -(ethy cyanoacrylate)} (PEGECA) amphiphilic graft copolymer nanoparticles (PEGECAT NPs) were prepared by the nanoprecipitation technique. The effects of solvent property, temperature, copolymer composition, and drug feeding on the drug-loaded amount and size of PEGECAT NPs were investigated. The morphological structure of PEGECAT NPs was characterized by transmission electron microscopy (TEM), proton nuclear magnetic resonance (1H NMR), and the size was measured by laser particle size analyzer (LPSA). In vitro release behaviors of drug from PEGECAT NPs were examined by high-pressure liquid chromatography (HPLC). The results demonstrate that PEGECAT NPs take on a spherical morphology with an inner core and outer shell before and after in vitro release. THP can be incorporated into the hydrophobic core of PEGECAT NPs and the drug-loaded amount is higher than 5%. The release of THP from PEGECAT NPs is initially fast and then slows down. The accumulated release is lower than 40% after 48 h. © 2008 Wiley Periodicals, Inc. J Appl Polym Sci, 2008 [source] Local application of n,3 or n,6 polyunsaturated fatty acids in the treatment of human experimental gingivitisJOURNAL OF CLINICAL PERIODONTOLOGY, Issue 4 2002Jörg Eberhard Abstract Background: Polyunsaturated fatty acids have the potential to attenuate inflammation by the synthesis of mediators of the 15-lipoxygenase pathways, which show opposite effects to the pro-inflammatory arachidonic acid metabolites such as leukotriene B4 (LTB4). Aims: The aim of this clinical study was to evaluate the effects of topical application of n,6 or n,6 polyunsaturated fatty acids in patients with experimental gingivitis. Methods: In each subject, similar teeth served as experimental and control over a 21-day non-hygiene phase and a 9-day resolving phase. Efficacy assessment was based on the bleeding on probing frequency (BOP) and the gingivocrevicular fluid volume (GCF). GCF was determined by inserting a filter paper strip for 30 s and measurements were performed on a Periotron 8000. The LTB4 concentration was analyzed by reversed-phase high-pressure liquid chromatography. Results: After 21 days of plaque growth, the BOP, GCF and LTB4 levels were significantly increased in all groups, with no differences between the control and experimental side. Rinsing of an area with established gingivitis for a 9-day period significantly reduced the GCF in the n,6 group (71.9 (18.7) versus 47.4 (11.4) Periotron Units, median (inter quartile range)). Conclusion: The topical application of n,6 or n,6 fatty acids failed to inhibit the development of experimental gingivitis. Rinsing with n,6 fatty acids could reduce the level of GCF in established experimental gingivitis. Zusammenfassung Hintergrund: Vielfach ungesättigte Fettsäuren haben das Potential, die Entzündung durch die Synthese von Mediatoren des 15-Lipoxygenaseweges zu behindern. Dies zeigt Gegeneffekte zu den pro-inflammatorischen Arachnoidonsäuremetaboliten wie Leukotrien B4 (LTB4). Ziele: Das Ziel dieser klinischen Studie war die Überprüfung des Effektes einer topischen Applikation von n,3 oder n,6 vielfach ungesättigten Fettsäuren bie Patienten mit experimenteller Gingivitis. Methoden: Bei jeder Person dienten ähnliche Zähne als Experiment und Kontrollen über eine 21tägige Nichthygiene-Phase und einer 9tägigen Erholungsphase. Wirksamkeitsmessungen basierten auf der Häufigkeit von Provokationsblutung (BOP) und dem Volumen der gingivalen krevikulären Flüssigkeit (GCF). GCF wurde durch Einbringen von Filterpapierstreifen für 30 Sekunden bestimmt. Die Messungen wurden mit einen Periotron 8000 durchgeführt. Die LTB4 Konzentration wurde mit der Umkehrphasen-Hochdruck-Flüssigkeitschromatographie analysiert. Ergebnisse: Nach 21 Tagen des Plaquewachstums waren die Level für BOP, GCF und LTB4 in allen Gruppen signifikant erhöht, ohne Differenzen zwischen den Kontrollen und den experimentellen Flächen. Die Spülung eines Gebietes mit etablierter Gingivitis für eine 9tägige Periode reduzierte die GCF in der n,6 Gruppe signifikant (71.9 (18.7) versus 47.4 (11.4) Peritron-Einheiten, Median (Zwischenquartilstreuung)). Zusammenfassung: Die topische Applikation von n,3 oder n,6 Fettsäuren verhindert die Entwicklung einer experimentellen Gingivitis nicht. Die Spülung mit n,6 Fettsäure konnte den Level der GCF bei einer bestehenden experimentellen Gingivitis reduzieren. Résumé Origine: Les acides gras poly-insaturés ont le potentiel d'atténuer l'inflammation en synthétisant des médiateurs des voies de la lipoxygénase 15 qui montrent des effets opposés aux métabolites de l'acide arachidonique pro-inflammatoire comme la leucotriène B4 (LTB4). But: Le but de cette étude clinique a été d'évaluer les effets de l'application topique d'acide gras poly-insaturés n,3 ou n,6 chez des patients effectuant d'une gingivite expérimentale. Méthodes: Chez chaque sujet, des dents semblables ont servi de sites tests et contrôles durant une phase sans hygiène buccale de 21 jours et une phase de retour à la normale de 9 jours. L'efficacité a été mesurée sur base de la fréquence du saignement au sondage (BOP) et le volume de fluide gingivale (GCF). Le GCF a été déterminé en insérant des papiers filtres pendant 30 s et les mesures ont été lues à l'aide du Périotron 8000. La concentration de LTB4 a été analysée par chromotographie liquide à haute pression à phrase arrière. Résultats: Après 21 jours d'accumulation de plaque dentaire les niveaux de BOP, GCF et LTB4 ont augmenté significativement dans tous les groupes sans aucune différence entre les sites tests et contrôles. Le rinçage d'une zone avec gingivite établie durant une période de 9 journées diminuait les GCF dans le groupe n,6 (unités du Péritron 72 (médian 19) versus 47 (11)). Conclusion: L'application topique d'acide gras n,3 ou n,6 ne permettait pas d'inhiber le développement de la gingivite expérimentale. Le rinçage avec des acides gras n,6 pouvait réduire le niveau de GCF dans la gingivite expérimentale établie. [source] THERMAL DEGRADATION KINETICS OF SUCROSE, GLUCOSE AND FRUCTOSE IN SUGARCANE MUST FOR BIOETHANOL PRODUCTIONJOURNAL OF FOOD PROCESS ENGINEERING, Issue 5 2006J. NOLASCO JR. ABSTRACT Thermal degradation of sugars contained in sugarcane must (21.5°Brix, pH 6.14) was evaluated at temperatures of 110, 120, 130 and 140C, using the thermal-death-time tube method, determining remaining sugars by high-pressure liquid chromatography. The study analyzed thermal degradation kinetics of both the total reducing sugars (TRS) and glucose and fructose individually. All curves of remaining sugars presented strong nonlinearity, with initial shoulder and final tail adjusted by an extended logistic model that was adapted for two species for TRS, and a simple logistic model for the monosaccharides. It was shown that rate constants are influenced by temperature according to two irreconcilable methods: the Arrhenius and the Bigelow methods. Obtained activation energies for fructose and glucose were quite coincident, 140.37 and 140.23 kJ/mol, respectively. Thermal resistance parameters were 21.59 and 21.61C, respectively. Comparison of the rate constants revealed that fructose degraded approximately 9,10 times faster than glucose. [source] The Green Oat Story: Possible Mechanisms of Green Color Formation in Oat Products during CookingJOURNAL OF FOOD SCIENCE, Issue 6 2009D.C. Doehlert ABSTRACT:, Consumers occasionally report greenish colors generated in their oat products when cooking in tap water. Here we have investigated pH and ferrous (Fe2+) ion as possible mechanisms for this color change. Steel-cut oat groats can turn brown-green color when cooked in alkaline conditions (pHs 9 to 12). Extraction of this color with methanol, and high-pressure liquid chromatography indicated a direct association of this color with the phenolic acid or avenanthramide content of the oat. The presence of 50 mM NaHCO3 in water will cause oat/water mixtures to turn alkaline when cooked as CO2 is driven off, generating OH, ion. Although tap water rarely, if ever, contains so much bicarbonate, bicarbonate is used as a leavening agent in baking applications. Industrial interests using baking soda or alkaline conditions during oat processing should be aware of possible off color generation. We have also found that as little as 10 ppm Fe2+ will turn oat products gray-green when cooked. The aleurone stained darker than the starchy endosperm. Other divalent cations, such as Ca2+ or Mg2+ had no effect on cooked oat color. As much as 50 ppm Fe2+ may be found in freshly pumped well water, but Fe2+ reacts quickly with oxygen and precipitates as Fe(OH)3. Thus, some freshly pumped well water may turn oats green when cooked, but if the water is left under atmospheric conditions for several hours, no discoloration will appear in the cooked oats. [source] Safety of Polyethylene Terephthalate Food Containers Evaluated by HPLC, Migration Test, and Estimated Daily IntakeJOURNAL OF FOOD SCIENCE, Issue 6 2008H.-J. Park ABSTRACT:, A comparative high-pressure liquid chromatography (HPLC) analysis of monomers, terephthalic acid (TPA), isophthalic acid (IPA), and dimethyl terephthalate (DMT) from polyethylene terephthalate (PET) food containers was conducted. Monomer linearities and sensitivities were calibrated between established and novel HPLC analyses. Safety of PET containers was evaluated with newly established detection methods for TPA, IPA, and DMT. Migration of the 3 monomers into food simulants (water, 4% acetic acid, 20% alcohol, and n-heptane) from 56 PET containers collected from open markets was monitored. Migrated monomers were not detected over 0.1 ppm of detection limit. The corresponding estimated daily intake was measured to confirm the safety of these publicly available PET containers and to permit comparison to the specific migration limit of the European Union. The estimated daily intake of 3 monomers migrating from PET was 0.0384 mg/kg each. This represented only 0.6% of the European Union's specific migration limit, confirming the safety of the examined containers. [source] An investigation of the 125I-radioiodination of colchicine for medical purposesJOURNAL OF LABELLED COMPOUNDS AND RADIOPHARMACEUTICALS, Issue 1 2009K. M. El-Azony Abstract A procedure for radioiodination of colchicine with iodine-125 is carried out via an electrophilic substitution reaction. The reaction parameters studied were colchicine concentration, pH of the reaction mixture, reaction time, temperature, different oxidizing agents and different organic media to optimize the conditions for the labeling of colchicine and to obtain a high radiochemical yield of the 125I-colchicine (125I-Col). Using 3.7,MBq of Na125I, 1.25,mM of colchicine as substrate, 1.1,mM of chloramine-T (CAT) as oxidizing agent in ethanol at 60°C for 5,min, a maximum radiochemical yield of 125I-Col (60%) was obtained. The specific activity of 125I-Col obtained was 44.4,MBq/0.5,mmol, and the labeled compound was not completely separated and purified from Col by means of high-pressure liquid chromatography (HPLC), so the uncertainty in the purity may affect the distribution and clearance routes due to the expected competition between 125I-Col and Col. The biological distribution in normal mice indicates the suitability of radioiodinated colchicine for imaging of muscles. Copyright © 2008 John Wiley & Sons, Ltd. [source] Synthesis and preliminary biological evaluation of a 99mTc-labeled hypericin derivative as a necrosis avid imaging agentJOURNAL OF LABELLED COMPOUNDS AND RADIOPHARMACEUTICALS, Issue 1 2008Humphrey Fonge Abstract Mono-[123I]iodohypericin and mono-[123I]iodohypericin monocarboxylic acid are iodine-123-labeled hypericin derivatives which have shown great promise in preclinical studies as necrosis avid imaging agents in animal models of infarction. In view of the more attractive properties of a 99mTc-labeled hypericin derivative, we have synthesized a conjugate of protohypericin monocarboxylic acid with S -benzoylmercaptoacetyldiglycyl-diaminopentane in an overall yield of 15%. The conjugate was labeled with technetium-99m by exchange labeling at pH 10 in a labeling yield of 95% followed by photocyclization to yield 99mTc-mercaptoacetyldiglycyl-1,5-diaminopentylene hypericincarboxamide (99mTc-13). The negatively charged 99mTc-13 complex was purified by reversed phase high-pressure liquid chromatography and the log,P7.4 was determined to be 2.36. In normal NMRI mice, the complex showed slow hepatobiliary clearance while plasma clearance was rapid. The tracer was evaluated in rats with reperfused hepatic infarction by ex vivo autoradiography, gamma counting and histochemical techniques. Unlike the radioiodinated hypericin derivatives, the new tracer agent did not show preferential uptake in necrotic tissue on autoradiography and gamma counting techniques. Conjugation of hypericin with a 99mTc-chelate, resulting in a change in size, charge and lipophilicity, had a profound effect on the necrosis avidity of the tracer agent. The results show that 99mTc-13 is not suitable for imaging necrosis. Copyright © 2008 John Wiley & Sons, Ltd. [source] The synthesis of tritiated (R)-2-methoxy- N - n -propyl-nor-apomorphine (MNPA)JOURNAL OF LABELLED COMPOUNDS AND RADIOPHARMACEUTICALS, Issue 13 2007Jonas Malmquist Abstract A method for the preparation of [3H]-MNPA ((R)-2-methoxy- N - n -propyl-norapomorphine) has been developed addressing the regioisomer problem as well as the oxidation problem. (R)-2, 10, 11-trihydroxy- N - n -propyl-norapomorphine was protected with 10, 11-dibenzyl or 10, 11-acetonide. The pure precursor was then methylated using [3H]-methyliodide. The product was isolated after deprotection and high-pressure liquid chromatography (HPLC) purification. Ascorbic acid was used as an antioxidant in the HPLC eluent and the stock solution. Characterization of the intermediates and products with 3H- and 1H-NMR was performed. A specific activity of 3.1,TBq/mmol (83.8,Ci/mmol) and 98.9% purity was obtained. Copyright © 2007 John Wiley & Sons, Ltd. [source] Lateral parabrachial afferent areas and serotonin mechanisms activated by volume expansionJOURNAL OF NEUROSCIENCE RESEARCH, Issue 16 2008Lisandra Oliveira Margatho Abstract Recent evidence has shown that the serotonergic mechanism of the lateral parabrachial nucleus (LPBN) participates in the regulation of renal and hormonal responses to isotonic blood volume expansion (BVE). We investigated the BVE-induced Fos activation along forebrain and hindbrain nuclei and particularly within the serotonergic clusters of the raphé system that directly project to the LPBN. We also examined whether there are changes in the concentration of serotonin (5HT) within the raphé nucleus in response to the same stimulus. With this purpose, we analyzed the cells doubly labeled for Fos and Fluorogold (FG) following BVE (NaCl 0.15 M, 2 ml/100 g b.w., 1 min) 7 days after FG injection into the LPBN. Compared with the control group, blood volume-expanded rats showed a significant greater number of Fos-FG double-labeled cells along the nucleus of the solitary tract, locus coeruleus, hypothalamic paraventricular nucleus, central extended amygdala complex, and dorsal raphé nucleus (DRN) cells. Our study also showed an increase in the number of serotonergic DRN neurons activated in response to isotonic BVE. We also observed decreased levels of 5HT and its metabolite 5-hydroxyindoleacetic acid (measured by high-pressure liquid chromatography) within the raphé nucleus 15 min after BVE. Given our previous evidence on the role of the serotonergic system in the LPBN after BVE, the present morphofunctional findings suggest the existence of a key pathway (DRN-LPBN) that may control BVE response through the modulation of 5HT release. © 2008 Wiley-Liss, Inc. [source] Purification and identification of an impurity in bulk hydrochlorothiazideJOURNAL OF PHARMACEUTICAL SCIENCES, Issue 11 2001Xueguang Fang Abstract Hydrochlorothiazide (6-chloro-3,4-dihydro-2H-1,2,4-benzothiadiazine-7-sulfonamide 1,1-dioxide) (HCTZ) 1 is a widely used diuretic and anti-hypertensive. Recently, the Pharmeuropa recognized a new impurity initially thought to be an HCTZ dimer 6, which consists of the active drug (HCTZ) linked via the former ,-ring methylene to a known degradate, 5-chloro-2,4-disulfamylaniline 2. In an effort to meet a new requirement, an analytical high-pressure liquid chromatography method was developed that was selective and sensitive to the subject impurity. The impurity was concentrated and purified using a combination of solid phase extraction and reverse-phase high-pressure liquid chromatography. Subsequently, the impurity has been identified as a specific HCTZ-CH2 -HCTZ isomer utilizing a variety of analytical techniques, including hydrolysis, ultraviolet spectroscopy, liquid chromatography/mass spectrometry, and 1H and 13C nuclear magnetic resonance (NMR) spectroscopy. The data resulting from the application of these analytical techniques confirm the identity of the impurity as a methylene bridged pair of HCTZ molecules; however, a total of six possible isomers 7a,f exist because of the presence of three reactive amines/sulfonamides on each HCTZ molecule. One unique molecular structure (4-[{6-chloro-3,4,-dihydro-2H-1,2,4-benzothiadiazine-7-sulfonamide-1,1-dioxide}-methyl]-chloro-3-hydro-H-1,2,4-benzothiadiazine-7-sulfonamide-1,1-dioxide) 7f was identified using two-dimensional COSY, NOESY, and TOCSY 1H NMR experiments. © 2001 Wiley-Liss, Inc. and the American Pharmaceutical Association J Pharm Sci 90:1800,1809, 2001 [source] Validation of subcutaneous microdialysis sampling for pharmacokinetic studies of flurbiprofen in the ratJOURNAL OF PHARMACEUTICAL SCIENCES, Issue 11 2001François-xavier Mathy Abstract The objective of this study was to validate subcutaneous (sc) microdialysis sampling to study flurbiprofen pharmacokinetics and plasma protein binding in the awake freely moving rat. A linear microdialysis probe was manufactured using a Hemophane® hollow fiber which was tested in vitro and in vivo for the recovery of flurbiprofen and naproxen used as retrodialysis marker. Flurbiprofen was administered intraperitoneally and intravenously at a dose of 20 mg/kg in rats. In both cases, conventional blood sampling and sc microdialysis sampling were simultaneously performed. The microdialysates were analyzed on-line by high-pressure liquid chromatography. Naproxen, which was shown to have a similar in vivo loss by retrodialysis as flurbiprofen (71.5,±,0.9% and 71.0,±,0.8% respectively, n,=,3), was used to continuously monitor probe recovery. Concentration-dependent protein binding of flurbiprofen was demonstrated in vivo based on experiments with a simultaneous sc microdialysis and blood sampling. Values of unbound fraction were similar to those reported previously by intravenous microdialysis sampling, demonstrating that the sc unbound concentrations are very similar to those in the central compartment. There was no significant difference among pharmacokinetic parameters (AUC, CL, t1/2z, Vd) for total or unbound flurbiprofen determined after intraperitoneal and intravenous administration. Subcutaneous microdialysis is a simple yet powerful tool to study the pharmacokinetics and the in vivo plasma protein binding of flurbiprofen in the awake unrestrained rat. © 2001 Wiley-Liss, Inc. and the American Pharmaceutical Association J Pharm Sci 90:1897,1906, 2001 [source] Effect of Oligosaccharide Alcohol Addition to Alumina Slurry and Translucent Alumina Produced by Slip CastingJOURNAL OF THE AMERICAN CERAMIC SOCIETY, Issue 5 2003Yuji Hotta A slurry used to produce dense green compacts by slip casting should exhibit low viscosity, high solids content, and good dispersion. Slurries with good characteristics were produced in the present study by adding oligosaccharide alcohol to an Al2O3 slurry with an NH4+ salt of poly(methacrylic acid) (NH4+ -PMA). The role of NH4+ -PMA and oligosaccharide alcohol in the Al2O3 slurry was examined by DTA, ,-potential measurement, high-pressure liquid chromatography, and viscometry. The viscosity of the slurry with NH4+ -PMA and oligosaccharide alcohol was lower than that of the slurry with NH4+ -PMA at a high solids content. Oligosaccharide alcohol did not interact with the Al2O3 surface. However, the Al2O3 slurry with NH4+ -PMA was influenced by the addition of oligosaccharide alcohol. We found that the dispersibility of the slurry was greatly improved by adding oligosaccharide alcohol. The transmittance of the Al2O3 ceramics produced by slip casting using the slurry with both NH4+ -PMA and oligosaccharide alcohol was higher than that of ceramics produced by slip casting using the slurry with NH4+ -PMA alone. The increased optical property resulted from low viscosity, which was attributed to the addition of oligosaccharide alcohol, at a high solids content. [source] Lactobacillus GG Does Not Affect D-Lactic Acidosis in Diarrheic Calves, in a Clinical SettingJOURNAL OF VETERINARY INTERNAL MEDICINE, Issue 3 2006Julia B. Ewaschuk D-lactate, produced by gastrointestinal fermentation, is a major contributor to metabolic acidosis in diarrheic calves. Lactobacillus rhamnosus GG survives gastrointestinal transit in the neonatal calf and does not produce D-lactate. To determine whether this probiotic reduces gastrointestinal D-lactate production or severity of diarrhea or both, 48 calves (mean, 11 days old; range, 2,30 days) admitted to the clinic for treatment of diarrhea were randomly allocated to 2 groups. The experimental group was given Lactobacillus rhamnosus GG (1×1011 cfu/d) PO, dissolved in milk or oral electrolyte solution, in addition to clinic treatment protocols; the other group served as a control. Serum and fecal samples were obtained at admission and at 24 and 48 hours after initial administration of Lactobacillus rhamnosus GG. All samples were analyzed for D-and L-lactate by using high-pressure liquid chromatography. Feces were also analyzed for pathogens, Lactobacillus rhamnosus GG recovery, and dry matter. D-lactic acidemia (>3 mmol/L) was present in 37/48 calves at admission. Lactobacillus rhamnosus GG was recovered in the feces of 13 experimental calves and 0 control calves 24 hours after administration. No difference in serum or fecal D- or L-lactate between the groups was detected at any time point. After therapy, D-lactic acidosis was absent at 48 hours in all but 1 calf. No relation between fecal pathogen (viral, bacterial, or protozoal) and degree of D-lactic acidosis was observed. The reduction in mortality and greater fecal dry matter in Lactobacillus rhamnosus GG-treated calves was not statistically significant. [source] Pharmacokinetics of 4-aminopyridine derivatives in dogsJOURNAL OF VETERINARY PHARMACOLOGY & THERAPEUTICS, Issue 5 2009N. J. OLBY Blockade of potassium channels with 4-aminopryidine (4-AP) restores conduction to demyelinated axons and improves function. Unfortunately, 4-AP causes adverse effects and its clinical effects are unpredictable and limited. Derivatives of 4-AP have been tested in models of spinal cord injury in guinea pigs; three derivatives (methyl-, ethyl- and t -butyl carbamate derivatives) showed promise. This study investigates the safety and pharmacokinetics of these derivatives in dogs. Each derivative was administered orally to dogs starting at doses below effective doses in guinea pigs, and increasing the dose on sequential days. Routine blood work was performed prior to and 24 h after drug administration, blood samples were collected at intervals over 24 h after drug administration, and dogs were monitored for side effects. Derivative plasma levels were determined using high-pressure liquid chromatography. Cerebrospinal fluid (CSF) samples were taken to determine CSF levels. No adverse effects were seen even when using doses higher than those that improved conduction in spinal cord injured guinea pigs. Peak plasma levels occurred at 36.6 (ethyl), 87 (t -butyl) and 175 (methyl) min and plasma level was related to drug dose. Penetration of the central nervous system (CNS) was good, with CSF levels higher than plasma levels for the t -butyl derivative. [source] Clinical efficacy of intravenous administration of marbofloxacin in a Staphylococcus aureus infection in tissue cages in poniesJOURNAL OF VETERINARY PHARMACOLOGY & THERAPEUTICS, Issue 6 2006M. VOERMANS Tissue cages (TC), implanted subcutaneously in the neck in eight ponies, were inoculated with Staphylococcus aureus (S. aureus) to determine the clinical efficacy of marbofloxacin in the treatment of this infection. From 21 h after inoculation, marbofloxacin (6 mg/kg) was administered intravenously (i.v.) once daily for 7 days. Samples of the tissue cage fluid (TCF) were taken to determine marbofloxacin concentrations (days 1, 3 and 7), using high-pressure liquid chromatography, and numbers of viable bacteria [colony forming units (CFU)] (days 1, 3, 7, 14 and 21). Statistical analysis was used to compare CFU before and after treatment. Clinical signs and CFU were used to evaluate the efficacy of treatment. Although, there was a slight decrease in CFU in all TC initially, the infection was not eliminated by marbofloxacin treatment in any of the ponies and abscesses formed. As the MIC (0.25 ,g/mL) did not change during treatment and the concentration of marbofloxacin during treatment (mean concentration in TCF was 0.89 ,g/mL on day 1, 0.80 ,g/mL on day 3 and 2.77 ,g/mL on day 7) was above MIC, we consider that the treatment failure might be attributable to the formation of a biofilm by S. aureus. Based on the present results, i.v. administration of marbofloxacin alone is not suitable for the elimination of S. aureus infections from secluded sites. [source] Presence of methylated arginine derivatives in orthotopic human liver transplantation: Relevance for liver functionLIVER TRANSPLANTATION, Issue 1 2003Paloma Martín-Sanz Orthotopic liver transplantation (OLT) is a frequent option in the treatment of liver diseases. During the cold ischemia period of the donor liver, there is an accumulation of metabolites that are potent inhibitors of the cytokine-inducible and endothelial nitric oxide synthase isoenzymes. We identified the presence of L - N -monomethylarginine and asymmetric dimethylarginine (ADMA) as the main inhibitors by means of analytic high-pressure liquid chromatography and mass spectrometry techniques. An average ADMA concentration of 450 ,mol/L was measured in the preservation medium of donor livers with poor outcomes after OLT. A statistically significant relationship was observed between the concentration of methylated arginine derivatives in the graft and liver function after OLT. These data suggest that measurement of methylated arginine, released after liver protein catabolism, might provide an indication of functional status of the liver that can help the development of strategies intended to improve graft viability. [source] The GABAergic-like system in the marine demosponge Chondrilla nuculaMICROSCOPY RESEARCH AND TECHNIQUE, Issue 11 2007Paola Ramoino Abstract Gamma-amino butyric acid (GABA) is believed to be the principal inhibitory neurotransmitter in the mammalian central nervous system, a function that has been extended to a number of invertebrate systems. The presence of GABA in the marine demosponge Chondrilla nucula was verified using immunofluorescence detection and high-pressure liquid chromatography. A strong GABA-like immunoreactivity (IR) was found associated with choanocytes, exopinacocytes, endopinacocytes lining inhalant, and exhalant canals, as well as in archaeocytes scattered in the mesohyl. The capacity to synthesize GABA from glutamate and to transport it into the vesicles was confirmed by the presence in C. nucula of glutamate decarboxylase (GAD) and vesicular GABA transporters (vGATs), respectively. GAD-like and vGAT-like IR show the same distribution as GABA-like IR. Supporting the similarity between sponge and mammalian proteins, bands with an apparent molecular weight of about 65,67 kDa and 57 kDa were detected using antibodies raised against mammalian GAD and vGAT, respectively. A functional metabotropic GABAB -like receptor is also present in C. nucula. Indeed, both GABAB R1 and R2 isoforms were detected by immunoblot and immunofluorescence. Also in this case, IR was found in choanocytes, exopinacocytes, and endopinacocytes. The content of GABA in C. nucula amounts to 1225.75 ± 79 pmol/mg proteins and GABA is released into the medium when sponge cells are depolarized. In conclusion, this study is the first indication of the existence of the GABA biosynthetic enzyme GAD and of the GABA transporter vGAT in sponges, as well as the first demonstration that the neurotransmitter GABA is released extracellularly. Microsc. Res. Tech., 2007. © 2007 Wiley-Liss, Inc. [source] Role of PAX2 gene polymorphisms in Henoch,Schonlein purpura nephritisNEPHROLOGY, Issue 1 2006ZHU-WEN YI SUMMARY: Objective: To investigate the distribution of polymorphisms in the PAX2 gene in children with Henoch,Schonlein purpura with and without nephritis (HSPN and HSP, respectively), with particular attention to the relationship between PAX2 gene polymorphisms and the development of kidney pathology. Methods: Genomic DNA was extracted from the peripheral leukocytes of 39 HSPN patients, 23 HSP patients without nephritis and 100 normal children, and three known single nucleotide polymorphisms (SNP), including 1410C>T, 1521A>C and 1544C>T in exon 8 and exon 9 of the PAX2 gene were studied as the candidate polymorphisms. The above two exons were amplified, the polymerase chain reaction (PCR) products were detected by denatured high-pressure liquid chromatography and direct DNA sequencing was performed for sequences with abnormal elution peaks. Results: In all samples confirmed by direct sequencing, we identified two SNP, which present as complete linkage haplotype 1410C>T + 1521A>C, in exon 8. We did not identify any SNP in exon 9. The frequency of the PAX2 heterozygous genotype 1410CT/1521AC in the HSPN group (28.20%) was significantly higher than in the HSP without HSPN group (4.35%) or in the control group (12.00%) (P < 0.05). The odds ratio (OR) values for HSPN and HSP were 6.05 and 2.62, respectively, and the 95% confidence intervals (CI) were 1.23,29.78 and 1.09,6.30, respectively. However, no differences in the frequency distribution was found between the HSP without nephritis and normal groups. Furthermore, there was no significant correlation between the polymorphism and clinical manifestation or kidney pathology in the HSPN group (P > 0.05). Conclusion: The 1410CT/1521AC PAX2 genotype does not increase susceptibility for HSP, but is likely to increase the susceptibility of kidney involvement, resulting in a HSPN diagnosis. [source] Speciation and distribution of arsenic and localization of nutrients in rice grainsNEW PHYTOLOGIST, Issue 1 2009E. Lombi Summary ,,Arsenic (As) contamination of rice grains and the generally low concentration of micronutrients in rice have been recognized as a major concern for human health. Here, we investigated the speciation and localization of As and the distribution of (micro)nutrients in rice grains because these are key factors controlling bioavailability of nutrients and contaminants. ,,Bulk total and speciation analyses using high-pressure liquid chromatography (HPLC),inductively coupled plasma mass spectrometry (ICP-MS) and X-ray absorption near-edge spectroscopy (XANES) was complemented by spatially resolved microspectroscopic techniques (µ-XANES, µ-X-ray fluorescence (µ-XRF) and particle induced X-ray emission (PIXE)) to investigate both speciation and distribution of As and localization of nutrients in situ. ,,The distribution of As and micronutrients varied between the various parts of the grains (husk, bran and endosperm) and was characterized by element-specific distribution patterns. The speciation of As in bran and endosperm was dominated by As(III),thiol complexes. ,,The results indicate that the translocation from the maternal to filial tissues may be a bottleneck for As accumulation in the grain. Strong similarities between the distribution of iron (Fe), manganese (Mn) and phosphorus (P) and between zinc (Zn) and sulphur (S) may be indicative of complexation mechanisms in rice grains. [source] | ||||||||||||||||||||||||||||||||||