Home About us Contact
|
Home About us Contact | ||
|
|
||
Thiol Levels (thiol + level)
Selected AbstractsQuantitation of reduced glutathione and cysteine in human immunodeficiency virus-infected patientsELECTROPHORESIS, Issue 10-11 2004Elena Sbrana Abstract Plasma viral load (VL) values and CD4+ cell count are employed clinically for initiation of therapy in the treatment of patients infected with human immunodeficiency virus (HIV), as previous clinical studies have shown a marked prevalence of acquired immunodeficiency sydrome (AIDS) development in seropositive individuals with VL values over 30,000 copies/mL. Many studies have shown that reduced glutathione (GSH) and cysteine (Cys) deficiency play an important role in the infection. We have developed capillary zone electrophoresis (CZE)-based assays and have used them to investigate the relationship between plasma and intracellular thiol levels and HIV-1 viremia in plasma. Blood samples from healthy volunteers and seropositive patients undergoing different antiretroviral regimes were analyzed in the study. The VL assay was based on CZE-UV detection of viral RNA at 260 nm. Determination of endogenous reduced Cys and GSH was achieved by CZE-UV detection of their mercurial complexes at 200 nm. We found that a decrease in GSH and Cys levels may be associated with disease progress. In fact, reduced GSH and Cys levels appear progressively reduced with increasing VL. [source] Adverse effects of arecoline and nicotine on human periodontal ligament fibroblasts in vitroJOURNAL OF CLINICAL PERIODONTOLOGY, Issue 3 2001Yu-Chao Chang Abstract Background, aims: The habit of betel nut chewing impinges on the daily lives of approximately 200 million people. Betel quid chewers have a higher prevalence of periodontal diseases than non-chewers. This study examined the pathobiological effects of arecoline, a major component of the betel nut alkaloids, on human periodontal ligament fibroblasts (PDLF) in vitro. Method: Cell viability, proliferation, protein synthesis, and cellular thiol levels were used to investigate the effects of human PDLF exposed to arecoline levels of 0 to 200 ,g/ml. In addition, nicotine was added to test how it modulated the effects of arecoline. Results: Arecoline significantly inhibited cell proliferation in a dose-dependent manner. At concentrations of 10 and 30 ,g/ml, arecoline suppressed the growth of PDLF by 20% and 50% (p<0.05), respectively. Arecoline also decreased protein synthesis in a dose-dependent manner during a 24-h culture period. A 100 ,g/ml concentration level of arecoline significantly inhibited protein synthesis to only 50% of that in the untreated control (p<0.05). Moreover, arecoline significantly depleted intracellular thiols in a dose-dependent manner. At concentrations of 25 ,g/ml and 100 ,g/ml, arecoline depleted about 18% and 56% of thiols (p<0.05), respectively. This suggests that arecoline itself might augment the destruction of periodontium associated with betel nut use. Furthermore, the addition of nicotine acted with a synergistic effect on the arecoline-induced cytotoxicity. At a concentration of 60 ,g/ml, arecoline suppressed the growth of PDLF by about 33%, and 5 mM nicotine enhanced the arecoline-induced cytotoxic response to cause about 66% cell death. Conclusion: During thiol depletion, arecoline may render human PDLF more vulnerable to reactive agents within cigarettes. Taken together, people who combine habits of betel nut chewing with cigarette smoking could be more susceptible to periodontium damage than betel nut chewing alone. Zusammenfassung Zielsetzung: Das Kauen von Betelnüssen gehört zum Alltag von ungefähr 200 Millionen Menschen. Betelnußkauer weisen eine höhere Prävalenz von Parodontalerkrankungen auf als Personen, die keine Betelnüsse konsumieren. In dieser Studie sollte der pathobiologische Effekt des Arekolins, das die Hauptkomponente des Betelnußalkaloides darstellt, auf menschliche Desmodontalfibroblasten (PDLF) in vitro untersuchen. Material und Methoden: Zellvitalität, Proliferationsrate, Proteinsynthese und zelluläre Thiolspiegel wurden genutzt, um zu untersuchen, welche Auswirkungen eine Exposition der PDLF gegenüber Arekolinspiegeln von 0 bis 200 ,g/ml hat. Zusätzlich wurde Nikotin beigefügt, um festzustellen wie das Nikotin den Effekt des Arekolins beeinflußt. Ergebnisse: Arekolin hemmt die Zellproliferation signifikant in dosisabhängiger Weise. Bei Konzentrationen von 10 und 30 ,g/ml unterdrückt Arekolin das Wachstum der PDLF um 20% bzw. 50% (p<0.05). Arekolin unterdrückt ebenfalls dosisabhängig die Proteinsynthese während der 24-stündigen Kultivierungsperiode. Ein Arekolinspiegel von 100 ,g/ml reduzierte die Proteinsynthese auf 50% im Vergleich zur unbehandelten Kontrollkultur (p<0.05). Auch die intrazellulären Thiolspiegel wurden dosisabhängig reduziert. Bei Konzentrationen von 25 und 100 ,g/ml wurden die Thiolspiegel um 18% bzw. 56% reduziert (p<0.05). Bei einer Konzentration von 60 ,g/ml unterdrückte das Arekolin das PDLF-Wachstum um 33%. Die Zugabe von 5 mM Nikotin verstärkte die durch Arekolin induzierte zytotoxische Wirkung, so daß es zum Zelltot von 66% kam. Schlußfolgerungen: Es scheint, daß Arekolin selbst zu der Schädigung des Parodonts beiträgt, die der Betelnuß zugeschreiben wird. Außerdem deuten die Ergebnisse darauf hin, daß Personen, die Betelnußkauen mit Nikotinkonsum kombinieren, empfindlicher für Schädigungen des Parodonts sind als solche, die nur Betelnüsse kauen. Während der Inaktivierung des Thiols könnte das Arekolin PDLF verletzlicher für andere reaktive Substanzen wie Nikotin machen. Résumé L'habitude de mastiquer de la noix de betel affecte la vie quotidienne de près de 200 millions de personnes. Les mâcheurs de betel présentent une prévalence plus élevée de maladies parodontales. Cette étude examine les effets pathologiques de l'arécoline, un composant majeur des alcaloïdes de la noix de betel, sur des fibroblastes du ligament parodontal humain (PDLF) in vitro. La viabilité cellulaire, la prolifération, la synthèse protéique, et les niveaux cellulaires de thiol ont été utilisés pour observer les effets de l'exposition de PDLF humains à des taux d'arécoline de 0 à 200 ,g/ml. De plus, de la nicotine fut ajouté pour tester la façon dont cela modulait les effets de l'arécoline. L'arécoline inhibait significativement la prolifération cellulaire de façon dose dépendante. A des concentrations de 10 à 30 ,g/ml, l'arécoline supprime la croissance des fibroblastes par 20 et 50% (p<0.05), respectivement. L'arécoline dimunuait également la synthèse des protéines de façon dose dépendante pendant une période de culture de 24 h. Une concentration de 100 ,g/ml d'arécoline inhibit la synthèse protéique à seulement 50% de celle du groupe controle non traité (p<0.05). De plus, l'arécoline réduit les thiols intracellulaires de façon dose dépendante. A des concentrations de 25 ,g/ml et 100 ,g/ml, l'arécoline réduit environ 18 à 56% des thiols, respectivement (p<0.05). Cela suggère que l'arécoline, elle même, peut augmenter la destruction du parodonte en association avec l'utilisation de noix de betel. De plus, l'addition de nicotine entrainait un effet synergique sur la cytotoxicité induite par l'arécoline. A une concentration de 60 ,g/ml, l'arécoline supprimait la croissance des PDLF d'environ 33% et 5 mM de nicotine augmentait cette réponse cytotoxique induite par l'arécoline, jusqu'à entrainer 66% de morts cellulaires. Lors de la réduction des thiols, l'arécoline pourrait rendre les PDLF humains plus vulnérables à des agents réactifs entrant dans la composition des cigarettes. Pris ensemble, les gens qui combinent des habitudes de mastication de noix de betel et de tabagisme, pourrait être plus susceptibles à des dommages parodontaux, que les gens qui utiliserait uniquement la noix de betel, mais sans fumer. [source] Gastroprotective properties of Myristica malabarica against indometacin-induced stomach ulceration: a mechanistic explorationJOURNAL OF PHARMACY AND PHARMACOLOGY: AN INTERNATI ONAL JOURNAL OF PHARMACEUTICAL SCIENCE, Issue 11 2007Debashish Banerjee The healing activity of the methanol extract of the spice rampatri, Myristica malabarica, (RM) and omeprazole against indometacin-induced stomach ulceration has been studied in a mouse model. Treatment with RM (40 mg kg,1 per day) and omeprazole (3 mg kg,1 per day) for 3 days could effectively heal the stomach ulceration, as revealed from the ulcer indices and histopathological studies. Compared with the ulcerated group, treatment with RM and omeprazole for 3 days reduced the macroscopic damage score by approximately 72% and 76%, respectively (P < 0.001), establishing the efficacy of RM. The extent of ulcer healing offered by 3 days' treatment with RM or omeprazole was better than that observed with natural recovery over 5 and 7 days (P < 0.05). The healing capacities of RM and omeprazole could be attributed to their antioxidant activity as well as the ability to enhance the mucin content of the gastric tissues. Both drugs reduced lipid peroxidation (by 42,44%) and protein carbonyl content (by 34%), and augmented non-protein thiol levels beyond normal values. Furthermore, RM improved the mucin level beyond the normal value, while omeprazole restored it to near normalcy. [source] Repeated bouts of aerobic exercise lead to reductions in skeletal muscle free radical generation and nuclear factor ,B activationTHE JOURNAL OF PHYSIOLOGY, Issue 16 2008Susan V. Brooks Chronic exercise improves endurance and skeletal muscle oxidative capacity. Despite the potential importance of reactive oxygen species (ROS) generated during exercise as regulators of these adaptations, the effect of repeated bouts of aerobic exercise on ROS generation by skeletal muscles during contractions has not been examined. Our aim was to establish the impact of repeated treadmill running exercise on muscle ROS generation and activation of redox-sensitive transcription factors. Following 8 weeks of treadmill running, mice displayed an improvement in running speed that was associated with an enhanced ability of gastrocnemius (GTN) muscles to maintain force during a protocol of isometric contractions. In contrast to GTN muscles of cage-sedentary (Sed) mice, muscles from exercised (Exer) mice did not release superoxide or nitric oxide during the isometric contractions. For male mice, basal levels of nuclear factor ,B (NF,B) and activator protein-1 (AP-1) DNA binding were increased by treadmill running, and the contraction-induced activation of NF,B and AP-1 observed in muscles of Sed mice was absent in Exer muscles. Also in contrast to Sed muscles, Exer muscles displayed no reductions in glutathione or protein thiol levels in response to contraction. Our observations of decreases for Exer compared with Sed muscles in contraction-induced (i) ROS generation, (ii) activation of redox-sensitive signalling pathways, and (iii) ROS stress suggest that exercise conditioning enhances the ability of skeletal muscle to readily and rapidly detoxify ROS and/or reduces ROS generation, providing protection from ROS-induced damage and reducing signals that might act to mediate further unnecessary adaptations. [source] Identification of an arsenic tolerant double mutant with a thiol-mediated component and increased arsenic tolerance in phyA mutantsTHE PLANT JOURNAL, Issue 6 2007Dong-Yul Sung Summary A genetic screen was performed to isolate mutants showing increased arsenic tolerance using an Arabidopsis thaliana population of activation tagged lines. The most arsenic-resistant mutant shows increased arsenate and arsenite tolerance. Genetic analyses of the mutant indicate that the mutant contains two loci that contribute to arsenic tolerance, designated ars4 and ars5. The ars4ars5 double mutant contains a single T-DNA insertion, ars4, which co-segregates with arsenic tolerance and is inserted in the Phytochrome A (PHYA) gene, strongly reducing the expression of PHYA. When grown under far-red light conditions ars4ars5 shows the same elongated hypocotyl phenotype as the previously described strong phyA-211 allele. Three independent phyA alleles, ars4, phyA-211 and a new T-DNA insertion allele (phyA-t) show increased tolerance to arsenate, although to a lesser degree than the ars4ars5 double mutant. Analyses of the ars5 single mutant show that ars5 exhibits stronger arsenic tolerance than ars4, and that ars5 is not linked to ars4. Arsenic tolerance assays with phyB-9 and phot1/phot2 mutants show that these photoreceptor mutants do not exhibit phyA -like arsenic tolerance. Fluorescence HPLC analyses show that elevated levels of phytochelatins were not detected in ars4, ars5 or ars4ars5, however increases in the thiols cysteine, , -glutamylcysteine and glutathione were observed. Compared with wild type, the total thiol levels in ars4, ars5 and ars4ars5 mutants were increased up to 80% with combined buthionine sulfoximine and arsenic treatments, suggesting the enhancement of mechanisms that mediate thiol synthesis in the mutants. The presented findings show that PHYA negatively regulates a pathway conferring arsenic tolerance, and that an enhanced thiol synthesis mechanism contributes to the arsenic tolerance of ars4ars5. [source] Protective effects of N-acetyl- L -cysteine against acute carbon tetrachloride hepatotoxicity in ratsCELL BIOCHEMISTRY AND FUNCTION, Issue 1 2008Yu. Z. Maksimchik Abstract In recent years, N-acetyl- L -cysteine (NAC) has been widely investigated as a potentially useful protective and antioxidative agent to be applied in many pathological states. The aim of the present work was further evaluation of the mechanisms of the NAC protective effect under carbon tetrachloride-induced acute liver injuries in rats. The rat treatment with CCl4 (4,g/kg, intragastrically) caused pronounced hepatolysis observed as an increase in blood plasma bilirubin levels and hepatic enzyme activities, which agreed with numerous previous observations. The rat intoxication was accompanied by an enhancement of membrane lipid peroxidation (1.4-fold) and protein oxidative damage (protein carbonyl group and mixed protein-glutathione disulphide formations) in the rat liver. The levels of nitric oxide in blood plasma and liver tissue significantly increased (5.3- and 1.5-fold, respectively) as blood plasma triacylglycerols decreased (1.6-fold). The NAC administration to control and intoxicated animals (three times at doses of 150,mg/kg) elevated low-molecular-weight thiols in the liver. The NAC administration under CCl4 -induced intoxication prevented oxidative damage of liver cells, decreased membrane lipid peroxidation, protein carbonyls and mixed protein-glutathione disulphides formation, and partially normalized plasma triacylglycerols. At the same time the NAC treatment of intoxicated animals did not produce a marked decrease of the elevated levels of blood plasma ALT and AST activities and bilirubin. The in vitro exposure of human red blood cells to NAC increased the cellular low-molecular-weight thiol levels and retarded tert -butylhydroperoxide-induced cellular thiol depletion and membrane lipid peroxidation as well as effectively inhibited hypochlorous acid-induced erythrocyte lysis. Thus, NAC can replenish non-protein cellular thiols and protect membrane lipids and proteins due to its direct radical-scavenging properties, but it did not attenuate hepatotoxicity in the acute rat CCl4 -intoxication model. Copyright © 2007 John Wiley & Sons, Ltd. [source] Electrochemical Investigation of Strontium,Metallothionein Interactions , Analysis of Serum and Urine of Patients with OsteoporosisELECTROANALYSIS, Issue 3-5 2009Ivo Fabrik Abstract The main aim of this paper is to study interaction between strontium and metallothionein (MT), and to determine changes in strontium and thiols (MT, reduced glutathione, cysteine, and homocysteine) level in plasma, serum, and urine samples of patients treated with strontium ranelate (SrR). To investigate the interactions between MT and strontium(II) ions, adsorptive transfer stripping technique coupled with differential pulse voltammetry (DPV) the Brdicka reaction was employed. Besides standard Brdicka signals (Co, RS2Co, Cat1, Cat2, Cat3), we observed new signal related to Sr-MT interaction. Further we investigated the effect of various time of interaction, concentration of strontium(II) ions and temperature of supporting electrolyte on Brdicka signals. Optimal time of interaction was 240,s. Under temperature of supporting electrolyte 20,°C, we measured linear dependence of Cat3 signal height on strontium(II) ions concentration. After that we have investigated the possibility of strontium-MT interactions, we were interested in strontium, MT and low molecular mass thiols levels in serum and urine of patients treated with strontium(II) ions to cure osteoporosis. Strontium concentration determined by atomic absorption spectrometry was 55±5,,g/L before and 10,500±1,400,,g/L at the 30th day of SrR administration. Levels of metallothionein in serum ranged from 0.1 to 6.4,,M. Correlation between serum strontium concentration and MT level was determined and correlation coefficient was R=0.93. [source] A new HPLC method for serum neopterin measurement and relationships with plasma thiols levels in healthy subjectsBIOMEDICAL CHROMATOGRAPHY, Issue 6 2004Ciriaco Carru Abstract Neopterin, a pyrazinopyrimidine compound, serves as a marker of cellular immune system activation, and it can be used as a prognostic predictor for certain types of diseases. We propose a new simple HPLC method to measure serum neopterin with highly sensitive ,uorimetric detection. After TCA serum protein precipitation, the supernatant was diluted ,ve times, injected into a C18 reversed-phase column and eluted at a ,ow rate of 1.5 mL/min by an isocratic water,acetonitrile (99:1) mobile phase. The natural ,uorescence of the molecule was detected at excitation wavelength 353 nm and emission 438 nm. In these conditions the neopterin retention time was about 4 min. Our proposed method was compared with a validated chromatographic separation, and the obtained data of the serum neopterin from 35 healthy volunteers were analysed by Passing,Bablok regression and Bland,Altman test. Neopterin measurement in healthy subjects was also employed to investigate on its potential relationships with plasma thiols levels. Copyright © 2004 John Wiley & Sons, Ltd. [source] | ||||||||||||||||