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1.
A selective determination of copper ions in water samples based on the fluorescence quenching of thiol‐capped CdTe quantum dots
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CdTe quantum dots (QDs) capped with different stabilizers, i.e. thioglycolic acid (TGA), 3‐mercaptopropionic acid (MPA) and glutathione (GSH) were investigated as fluorescent probes for the determination of Cu2+. The stabilizer was shown to play an important role in both the sensitivity and selectivity for the determination of Cu2+. TGA‐capped CdTe QDs showed the highest sensitivity, followed by the MPA and GSH‐capped CdTe QDs, respectively. The TGA‐ and MPA‐capped CdTe QDs were not selective for Cu2+ that was affected by Ag+. The GSH‐capped CdTe QDs were insensitive to Ag+ and were used to determine Cu2+ in water samples. Under optimal conditions, quenching of the fluorescence intensity (F0/F) increased linearly with the concentration of Cu2+ over a range of 0.10–4.0 µg/mL and the detection limit was 0.06 µg/mL. The developed method was successfully applied to the determination of Cu2+ in water samples. Good recoveries of 93–104%, with a relative standard deviation of < 6% demonstrated that the developed simple method was accurate and reliable. The quenching mechanisms were also described. Copyright © 2015 John Wiley & Sons, Ltd. 相似文献
2.
Ferrous ion (Fe2+) has been considered to be a cause of neuronal oxidative injury. Since body fluids contain protein and serum is an essential component of tissue culture medium, we have examined the role of serum protein on Fe2+-mediated oxidative stress using PC12 cells and rat cerebral cortices. Fe2+ or the combination of ascorbate and Fe2+ increased concentrations of thiobarbituric acid reactive substances (TBARS) in PC12 cells and cerebrocortical homogenates in medium (RPMI 1640), but did not increase TBARS when the medium was supplemented with 10% fetal bovine serum. Treatment with ascorbate/Fe2+ in serum-free medium reduced endogenous glutathione (GSH) concentration in PC12 cells. However, the medium supplemented with serum did not reduce GSH concentrations. PC12 cell death induced by ascorbate/Fe2+ was alleviated by increasing serum or bovine albumin concentrations in the medium. These observations indicated that oxidative injury caused by the transition metal ion could be lessened by adding fetal bovine serum to culture medium. 相似文献
3.
Santos LO Gonzales TA Ubeda BT Monte Alegre R 《Applied microbiology and biotechnology》2007,77(4):763-769
A strategy of experimental design using a fractional factorial design (FFD) and a central composite rotatable design (CCRD)
were carried out with the aim to obtain the best conditions of temperature (20–30°C), agitation rate (100–300 rpm), initial
pH (5.0–7.0), inoculum concentration (5–15%), and glucose concentration (30–70 g/l) for glutathione (GSH) production in shake-flask
culture by Saccharomyces cerevisiae ATCC 7754. By a FFD (25–2), the agitation rate, temperature, and pH were found to be significant factors for GSH production. In CCRD (22) was obtained a second-order model equation, and the percent of variation explained by the model was 95%. The results showed
that the optimal culture conditions were agitation rate, 300 rpm; temperature, 20°C; initial pH, 5; glucose, 54 g/l; and inoculum
concentration, 5%. The highest GSH concentration (154.5 mg/l) was obtained after 72 h of fermentation. 相似文献
4.
Personal model‐assisted identification of NAD+ and glutathione metabolism as intervention target in NAFLD
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Adil Mardinoglu Elias Bjornson Cheng Zhang Martina Klevstig Sanni Söderlund Marcus Ståhlman Martin Adiels Antti Hakkarainen Nina Lundbom Murat Kilicarslan Björn M Hallström Jesper Lundbom Bruno Vergès Peter Hugh R Barrett Gerald F Watts Mireille J Serlie Jens Nielsen Mathias Uhlén Ulf Smith Hanns‐Ulrich Marschall Marja‐Riitta Taskinen Jan Boren 《Molecular systems biology》2017,13(3)
To elucidate the molecular mechanisms underlying non‐alcoholic fatty liver disease (NAFLD), we recruited 86 subjects with varying degrees of hepatic steatosis (HS). We obtained experimental data on lipoprotein fluxes and used these individual measurements as personalized constraints of a hepatocyte genome‐scale metabolic model to investigate metabolic differences in liver, taking into account its interactions with other tissues. Our systems level analysis predicted an altered demand for NAD+ and glutathione (GSH) in subjects with high HS. Our analysis and metabolomic measurements showed that plasma levels of glycine, serine, and associated metabolites are negatively correlated with HS, suggesting that these GSH metabolism precursors might be limiting. Quantification of the hepatic expression levels of the associated enzymes further pointed to altered de novo GSH synthesis. To assess the effect of GSH and NAD+ repletion on the development of NAFLD, we added precursors for GSH and NAD+ biosynthesis to the Western diet and demonstrated that supplementation prevents HS in mice. In a proof‐of‐concept human study, we found improved liver function and decreased HS after supplementation with serine (a precursor to glycine) and hereby propose a strategy for NAFLD treatment. 相似文献
5.
To generate an industrial strain of Hansenula polymorpha capable of yielding greater levels of glutathione (GSH), wild strain H. polymorpha DL-1 cells were mutated using a nitrogen ion beam, a novel mutagen. At an energy level of 20 keV and dose of 2.13 × 1016 ions/cm2, H. polymorpha strain 28 (HP28) with a high-yield of GSH was screened. HP28 intracellular GSH levels reached 337.16 mg/L by ion beam implantation, 1.56 times greater than that of the wild type strain when the fermentation time was shortened from 48 hr to 42 hr, greatly improving efficiency and reducing the cost of industrial-scale production. The enhanced efficiency of HP28 is promising for GSH production from lignocellulosic materials. Therefore, the ion beam implantation would be a cost-effective alternative to the conventional mutation method for engineering yeast and improving its utility. 相似文献
6.
We examined the brain oxidative stress which accompanies 30 min of bilateral carotid artery ligation (BCAL) in terms of changes in brain levels of glutathione; reduced (GSH) and oxidized (GSSG) forms and the exacerbation of oxidative stress by disulfiram (DSF). These results indicate that BCAL alone decreases GSH content and limits glutathione reductase (GR) activity, and these changes were enhanced by DSF pretreatment. Similar observations were recorded with DSF alone. GR activity (74.3±4.0 µmol min–1 mg–1 tissue; p<0.001) and GSH content (1.23±0.06 µmol min–1g–1 tissue; p<0.001) was attenuated in rats subjected to synergistic effect of BCAL and DSF with a concomitant increase of GSSG (0.006±0.006 µmol min–1 g–1 tissue; p<0.001). Recovery of GSH/GSSG level and GR activity during reperfusion following 30 min BCAL was considerably delayed (96 h) in the BCAL and DSF group as compared to the recovery time of 24 h in the group subjected to BCAL-reperfusion alone. Perturbation of GSH/GSSG homeostasis as a result of BCAL was augmented by DSF. These findings clearly demonstrate central nervous system oxidative stress due to a BCAL-DSF synergistic effect. Based on the results obtained with this model, we conclude that DSF increases brain oxidative stress and this may be detrimental to alcoholics who might drink and develop an acetaldehyde-induced hypotension while taking DSF. 相似文献
7.
The effect of glutathione (GSH) on the ultraviolet (UV) induction of lambda prophage was investigated in lysogenic Escherichia coli. The data showed that extracellular GSH could inhibit the UV induction of lambda prophage. The inhibitory rates were concentration
dependent, and the maximal rate obtained was 94% with 3.0 M GSH. The effect was also measured in three different lambda lysogens:
a wild-type strain (wt), an isogenic GSH-deficient strain, and an isogenic strain producing increased amounts of GSH. The
result showed that when subjected to UV irradiation (254 nm, 60 J m−2), GSH-deficient strain was approximately fivefold more sensitive to be lysed than wt, whereas the strain with higher intracellular
GSH levels was only 28% susceptible to be lysed. With electron spin resonance and spin trapping techniques, we observed that
free radical signals occurred in the suspensions of UV irradiated lysogenic cells and the intensity of signals was influenced
by GSH levels. These results indicate that GSH can significantly inhibit the UV induction of lambda prophage, and that this
effect is correlated to its capacity to scavenge free radicals generated after UV irradiation. 相似文献
8.
Yucheng Zhao Xiaohong Bian Xiaoqing You Fei Shao Xi Xiang Xuepeng Deng Ganggang Zhao Jiyang Xu 《Engineering in Life Science》2013,13(2):156-162
Glutathione (GSH), an important tripeptide compound, is widely used as a therapeutic and in the food and cosmetic industries. To improve its production yield, we added the antibiotic nystatin to a batch fermentation of Saccharomyces cerevisiae, at different concentrations and at various times. Based on the results that nystatin can effectively stimulate GSH accumulation but at the same time inhibits cell growth, a three‐point addition strategy (0.05 mg/L at 8 h, 0.25 mg/L at 16 h, and 0.5 mg/L at 20 h) was developed to maximize GSH production. As a result, a maximum yield of 237.8 mg/L was obtained, which was by 50.6% higher than without the addition of nystatin. When combining this strategy with cysteine addition, the GSH yield increased to 278.9 mg/L. Subsequently, the γ‐glutamylcysteine synthetase (γ‐GCS) activity and K+ concentration were analyzed to investigate the possible mechanism involved in the increased production. It was found that the nystatin‐induced increase in the GSH yield was associated with a higher γ‐GCS activity and K+ concentration. 相似文献
9.
Lagos CF Araya-Secchi R Thomas P Pérez-Acle T Tapia RA Salas CO 《Journal of molecular modeling》2012,18(5):2055-2064
Trypanosoma cruzi glutamate cysteine ligase (TcGCL) is considered a potential drug target to develop novel antichagasic drugs. We have used
a variety of computational methods to investigate the interactions between TcGCL with Glutathione (GSH). The three-dimensional
structure of TcGCL was constructed by comparative modeling methods using the Saccharomyces cerevisiae glutamate cysteine ligase as template. Molecular dynamics simulations were used to validate the TcGCL model and to analyze
the molecular interactions with GSH. Using RMSD clustering, the most prevalent GSH binding modes were identified paying attention
to the residues involved in the molecular interactions. The GSH binding modes were used to propose pharmacophore models that
can be exploited in further studies to identify novel antichagasic compounds. 相似文献
10.
Toyin Adeyemi Ogunrinu Harald Sontheimer 《The Journal of biological chemistry》2010,285(48):37716-37724
Glutathione (GSH) is an essential antioxidant responsible for the maintenance of intracellular redox homeostasis. As tumors outgrow their blood supply and become hypoxic, their redox homeostasis is challenged by the production of nitric oxide and reactive oxygen species (ROS). In gliomas, the sustained import of l-cystine via the l-cystine/l-glutamate exchanger, system xc−, is rate-limiting for the synthesis of GSH. We show that hypoxia causes a significant increase in NO and ROS but without affecting glioma cell growth. This is explained by a concomitant increase in the utilization of GSH, which is accompanied by an increase in the cell-surface expression of xCT, the catalytic subunit of system xc−, and l-cystine uptake. Growth was inhibited when GSH synthesis was blocked by buthionine sulfoximine (BSO), an inhibitor of the enzyme required for GSH synthesis, or when cells were deprived of l-cystine. These findings suggest that glioma cells show an increased requirement for GSH to maintain growth under hypoxic conditions. Therefore, approaches that limit GSH synthesis such as blocking system xc− may be considered as an adjuvant to radiation or chemotherapy. 相似文献
11.
Using the model of glutathione (GSH) depletion, possible role of GSH in the maintenance of blood-brain barrier (BBB) integrity was evaluated in rats. Administration (ip) of GSH depletors, diethyl maleate (DEM, 1–4 mmol/kg), phorone (2–3 mmol/kg) and 2-cyclohexene-1-one (CHX, 1 mmol/kg), to male adults was found to deplete brain and liver GSH and increase the BBB permeability to micromolecular tracers (sodium fluorescein and [14C]sucrose) in a dose-dependent manner at 2h. However, BBB permeability to macromolecular tracers such as horseradish peroxidase and Evan's blue remained unaltered. It was also shown that observed BBB permeability dysfunction was associated with brain GSH depletion. A lower magnitude of BBB increase in rat neonates, as compared to adults, indicated a possible bigger role of GSH in the BBB function of mature brain. The treatment with N-acetylcysteine, methionine and GSH provided a partial to full protection against DEM-induced brain (microvessel) GSH depletion and BBB dysfunction; however, the treatment with -tocopherol, ascorbic acid and turmeric were not effective. Our studies showed that cerebral GSH plays an important role in maintaining the functional BBB integrity. 相似文献
12.
Kai Li Yuan Chi Kun Gao Qiaojing Yan Hiroyuki Matsue Masayuki Takeda Masanori Kitamura Jian Yao 《PloS one》2013,8(2)
Background
Many signaling molecules and pathways that regulate gap junctions (GJs) protein expression and function are, in fact, also controlled by GJs. We, therefore, speculated an existence of the GJ channel-mediated self-regulation of GJs. Using a cell culture model in which nonjunctional connexin43 (Cx43) hemichannels were activated by cadmium (Cd2+), we tested this hypothesis.Principal Findings
Incubation of Cx43-transfected LLC-PK1 cells with Cd2+ led to an increased expression of Cx43. This effect of Cd2+ was tightly associated with JNK activation. Inhibition of JNK abolished the elevation of Cx43. Further analysis revealed that the changes of JNK and Cx43 were controlled by GSH. Supplement of a membrane-permeable GSH analogue GSH ethyl ester or GSH precursor N-acetyl-cystein abrogated the effects of Cd2+ on JNK activation and Cx43 expression. Indeed, Cd2+ induced extracellular release of GSH. Blockade of Cx43 hemichannels with heptanol or Cx43 mimetic peptide Gap26 to prevent the efflux of GSH significantly attenuated the Cx43-elevating effects of Cd2+.Conclusions
Collectively, our results thus indicate that Cd2+-induced upregulation of Cx43 is through activation of nonjunctional Cx43 hemichannels. Our findings thus support the existence of a hemichannel-mediated self-regulation of Cx43 and provide novel insights into the molecular mechanisms of Cx43 expression and function. 相似文献13.
Cabral CB Bullock KH Bischoff DJ Tompkins RG Yu YM Kelleher JK 《Analytical biochemistry》2008,379(1):40-44
Glutathione (GSH), an intracellular tripeptide that combats oxidative stress, must be continually replaced due to loss through conjugation and destruction. Previous methods, estimating the synthesis of GSH in vivo, used constant infusions of labeled amino acid precursors. We developed a new method based on incorporation of 2H from orally supplied 2H2O into stable C-H bonds on the tripeptide. The incorporation of 2H2O into GSH was studied in rabbits over a 2-week period. The method estimated N, the maximum number of C-H bonds in GSH that equilibrate with 2H2O as amino acids. GSH was analyzed by liquid chromatography/mass spectrometry after derivatization to yield GSH-N-ethylmaleimide (GSNEM). A model, which simulated the expected abundance at each mass isotopomer for the GSNEM ion at various values for N, was used to find the best fit to the data. The plateau labeling fit best a model with N = 6 of a possible 10 C-H bonds. Thus, the amino acid precursors do not completely equilibrate with 2H2O prior to GSH synthesis. Advantages of this new method include replacing costly amino acid infusions with the oral administration of 2H2O and a statistical basis for estimating N. 相似文献
14.
Hydrogen selenide ion (HSe?) has an important role in the metabolism of the essential trace element selenium. Several redox reactions of selenide were found to be dominated by the amount of colloidal elemental selenium (Se°) generated during the reaction. The following reaction of selenide with the disulfide, oxidized glutathione (GSSG), was used as an example: HSe? + GSSG + H+ → Se° + 2 GSH. The resulting thiol is reduced glutathione (GSH; γ-glutamylcysteinylglycine). By following this reaction with polarography, it was seen that the ratio of colloidal selenium produced to selenide unreacted was a constant 2.1 ± 0.1, and was the only factor found to determine the extent of oxidation. This is best explained by the hypothesis that freshly generated colloidal selenium adsorbs selenide readily; no evidence for polyselenide formation was found. Adsorption of selenide should be considered in any reaction involving the oxidation of selenide to colloidal selenium. 相似文献
15.
16.
Adducts of oxylipin electrophiles to glutathione reflect a 13 specificity of the downstream lipoxygenase pathway in the tobacco hypersensitive response
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Davoine C Falletti O Douki T Iacazio G Ennar N Montillet JL Triantaphylidès C 《Plant physiology》2006,140(4):1484-1493
The response to reactive electrophile species (RES) is now considered as part of the plant response to pathogen and insect attacks. Thanks to a previously established high-performance liquid chromatography tandem mass spectrometry methodology, we have investigated the production of oxylipin RES adducts to glutathione (GSH) during the hypersensitive response (HR) of plants. We have observed that RES conjugation to GSH in tobacco (Nicotiana tabacum) leaves is facile and nonspecific. In cryptogein-elicited tobacco leaves, we show that the oxylipin RES adducts to GSH are produced in correlation with GSH consumption, increase in glutathione S-transferase activity, and the appearance of the cell death symptoms. In this model, the adducts arise mainly from the downstream 13 lipoxygenase (LOX) metabolism, although the induced 9 LOX pathway leads massively to the accumulation of upstream metabolites. The main adducts were obtained from 2-hexenal and 12-oxo-phytodienoic acid. They accumulate transiently as 1-hexanol-3-GSH, a reduced adduct, and 12-oxo-phytodienoic acid-GSH, respectively. RES conjugation does not initiate cell death but explains part of the GSH depletion that accompanies HR cell death. The nature of these GSH conjugates shows the key role played by the 13 LOX pathway in RES signaling in the tobacco HR. 相似文献
17.
Dan Nguyen Susan L. Samson Vasumathi T. Reddy Erica V. Gonzalez Rajagopal V. Sekhar 《Aging cell》2013,12(3):415-425
Aging is associated with impaired fasted oxidation of nonesterified fatty acids (NEFA) suggesting a mitochondrial defect. Aging is also associated with deficiency of glutathione (GSH), an important mitochondrial antioxidant, and with insulin resistance. This study tested whether GSH deficiency in aging contributes to impaired mitochondrial NEFA oxidation and insulin resistance, and whether GSH restoration reverses these defects. Three studies were conducted: (i) in 82‐week‐old C57BL/6 mice, the effect of naturally occurring GSH deficiency and its restoration on mitochondrial 13C1‐palmitate oxidation and glucose metabolism was compared with 22‐week‐old C57BL/6 mice; (ii) in 20‐week C57BL/6 mice, the effect of GSH depletion on mitochondrial oxidation of 13C1‐palmitate and glucose metabolism was studied; (iii) the effect of GSH deficiency and its restoration on fasted NEFA oxidation and insulin resistance was studied in GSH‐deficient elderly humans, and compared with GSH‐replete young humans. Chronic GSH deficiency in old mice and elderly humans was associated with decreased fasted mitochondrial NEFA oxidation and insulin resistance, and these defects were reversed with GSH restoration. Acute depletion of GSH in young mice resulted in lower mitochondrial NEFA oxidation, but did not alter glucose metabolism. These data suggest that GSH is a novel regulator of mitochondrial NEFA oxidation and insulin resistance in aging. Chronic GSH deficiency promotes impaired NEFA oxidation and insulin resistance, and GSH restoration reverses these defects. Supplementing diets of elderly humans with cysteine and glycine to correct GSH deficiency could provide significant metabolic benefits. 相似文献
18.
Heavy metal-induced glutathione accumulation and its role in heavy metal detoxification in Phanerochaete chrysosporium 总被引:1,自引:0,他引:1
Piao Xu Liang Liu Guangming Zeng Danlian Huang Cui Lai Meihua Zhao Chao Huang Ningjie Li Zhen Wei Haipeng Wu Chen Zhang Mingyong Lai Yibin He 《Applied microbiology and biotechnology》2014,98(14):6409-6418
Phanerochaete chrysosporium are known to be vital hyperaccumulation species for heavy metal removal with admirable intracellular bioaccumulation capacity. This study analyzes the heavy metal-induced glutathione (GSH) accumulation and the regulation at the intracellular heavy metal level in P. chrysosporium. P. chrysosporium accumulated high levels of GSH, accompanied with high intracellular concentrations of Pb and Cd. Pb bioaccumulation lead to a narrow range of fluctuation in GSH accumulation (0.72–0.84 μmol), while GSH plummeted under Cd exposure at the maximum value of 0.37 μmol. Good correlations between time-course GSH depletion and Cd bioaccumulation were determined (R 2?>?0.87), while no significant correlations have been found between GSH variation and Pb bioaccumulation (R 2?<?0.38). Significantly, concentration-dependent molar ratios of Pb/GSH ranging from 0.10 to 0.18 were observed, while molar ratios of Cd/GSH were at the scope of 1.53–3.32, confirming the dominant role of GSH in Cd chelation. The study also demonstrated that P. chrysosporium showed considerable hypertolerance to Pb ions, accompanied with demand-driven stimulation in GSH synthesis and unconspicuous generation of reactive oxygen stress. GSH plummeted dramatically response to Cd exposure, due to the strong affinity of GSH to Cd and the involvement of GSH in Cd detoxification mechanism mainly as Cd chelators. Investigations into GSH metabolism and its role in ameliorating metal toxicity can offer important information on the application of the microorganism for wastewater treatment. 相似文献
19.
Renal cellular concentration of glutathione (GSH) increases after exposure to a subtoxic dose of inorganic mercury (Hg2+). In the present study, we tested the hypothesis that the increase in renal cellular concentration of GSH after exposure to a subtoxic dose of Hg2+ (0.5 μmol HgCl2/kg body wt) is due to induction of GSH synthesis. Rats were treated in vivo with HgCl2, and renal proximal tubular (PT) and distal tubular (DT) cells were isolated 24 hours later. PT cells were studied as the presumed target site for Hg2+, and DT cells were investigated as a nontarget cell population. γ-Glutamylcysteine synthetase activity increased after exposure to Hg2+ in PT cells when expressed on a per cell basis. Increases in activities of glutathione disulfide (GSSG) reductase, GSH peroxidase, and several enzymes involved in cellular energetics occurred after exposure to Hg2+. Many of these increases were observed in both PT and DT cells, indicating that the responses to Hg2+ were not restricted to the PT cells. These results are consistent with the hypothesis that in vivo exposure to a subtoxic dose of Hg2+ is also associated with induction of GSH synthesis and other key cellular enzymes. Early changes in GSH metabolism associated with exposure to Hg2+ appear to occur both in the primary target cell population and in more distal nephron sites. © 1996 John Wiley & Sons, Inc. 相似文献
20.