A novel derivatization approach for simultaneous determination of glyoxal, methylglyoxal, and 3-deoxyglucosone in plasma by gas chromatography–mass spectrometry

Wu, Meng-Yan; Chen, Bud-Gen; Chang, Chiung Dan; Huang, Mei-Han; Wu, Tai-Guang; Chang, Daw-Ming; Lee, Yau-Jiunn; Wang, Hsiang Ching; Lee, Chu-I; Chern, Chi-Liang; Liu, Ray H.

doi:10.1016/j.chroma.2008.07.040

Cited by 29 publications

(24 citation statements)

References 23 publications

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“…DNA extracted from parasites at selected time points [53] was hydrolyzed and dephosphorylated as described in [54]. The released deoxyribose moiety (250 µl) was oximated by addition of 0.01 M HCl (1.84 ml) and O-(2,3,4,5,6-pentafluorobenzyl) hydroxylamine acetate (20 µl, 25 mg/ml) and incubated (90°C, 3 h) [55]. Oximes were extracted in 2 steps using an ethyl acetate/hexane mix (1∶1 v/v) and pure ethyl acetate.…”

Section: Methodsmentioning

confidence: 99%

Induction of a Stringent Metabolic Response in Intracellular Stages of Leishmania mexicana Leads to Increased Dependence on Mitochondrial Metabolism

et al. 2014

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Leishmania parasites alternate between extracellular promastigote stages in the insect vector and an obligate intracellular amastigote stage that proliferates within the phagolysosomal compartment of macrophages in the mammalian host. Most enzymes involved in Leishmania central carbon metabolism are constitutively expressed and stage-specific changes in energy metabolism remain poorly defined. Using 13C-stable isotope resolved metabolomics and 2H2O labelling, we show that amastigote differentiation is associated with reduction in growth rate and induction of a distinct stringent metabolic state. This state is characterized by a global decrease in the uptake and utilization of glucose and amino acids, a reduced secretion of organic acids and increased fatty acid β-oxidation. Isotopomer analysis showed that catabolism of hexose and fatty acids provide C4 dicarboxylic acids (succinate/malate) and acetyl-CoA for the synthesis of glutamate via a compartmentalized mitochondrial tricarboxylic acid (TCA) cycle. In vitro cultivated and intracellular amastigotes are acutely sensitive to inhibitors of mitochondrial aconitase and glutamine synthetase, indicating that these anabolic pathways are essential for intracellular growth and virulence. Lesion-derived amastigotes exhibit a similar metabolism to in vitro differentiated amastigotes, indicating that this stringent response is coupled to differentiation signals rather than exogenous nutrient levels. Induction of a stringent metabolic response may facilitate amastigote survival in a nutrient-poor intracellular niche and underlie the increased dependence of this stage on hexose and mitochondrial metabolism.

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Section: Methodsmentioning

confidence: 99%

Induction of a Stringent Metabolic Response in Intracellular Stages of Leishmania mexicana Leads to Increased Dependence on Mitochondrial Metabolism

et al. 2014

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“…1 ml of the supernatant was derivatized by incubation with 100 μL O ‐(2,3,4,5,6‐pentafluorobenzyl) hydroxylamine (25 mg/mL) and 4.6 ml HCl (pH 3.7) at 50°C for 2 h. Solvent extraction was performed in 3 mL ether/hexane (1 : 1) and the organic phase was dried at 25°C under nitrogen. Upon reconstitution with 50 μL of ethyl acetate, gas chromatography‐mass spectrometry (GC‐MS) analysis was carried out as described previously (Wu et al. 2008).…”

Section: Methodsmentioning

confidence: 99%

Methylglyoxal‐mediated anxiolysis involves increased protein modification and elevated expression of glyoxalase 1 in the brain

Hambsch

Chen

Brenndörfer

et al. 2010

Journal of Neurochemistry

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To elucidate physiological mechanisms underlying pathological anxiety, valid animal models are essential (Cryan and Holmes 2005). Previous studies confirmed that the high anxiety-related behavior/low anxiety-related behavior (HAB/ LAB) mouse model for trait anxiety mimics many clinically relevant features typical of psychiatric patients (Krömer et al. 2005;Landgraf et al. 2007). In this model, behavioral phenotyping on the elevated-plus-maze (EPM) test was used as selection criterion for bidirectional intra-strain (CD1) breeding. Consequently, inbreeding for more than 30 generations created hyper-anxious (HAB) and hypo-anxious (LAB) mice. These animals provide robust phenotypes for extremes in anxiety-related behavior, and show high comorbidity with depression-like behavior. Inbreeding with focus on a particular trait included a strong accumulation of specific genetic material. Therefore, HAB/LAB animals serve as a potent model to analyze candidates underlying trait anxiety. One of the first protein markers shown to be differentially regulated in extremes of trait anxiety was glyoxalase1 (GLO1; lactoylglutathione lyase, EC 4.4.1.5).Mouse GLO1 is an enzyme of 21kDa, containing 184 amino acids that is found as a dimer in the cytosol of cells. Its physiological function is the catalytic conversion of Abbreviations used: AGE, advanced glycation end; ARMS, amplification refractory mutation system; CNV, copy number variant; enpp5, ectonucleotide pyrophosphatase/phosphodiesterase 5; EPM, elevatedplus-maze; GC-MS, gas chromatography-mass spectrometry; GLO1, glyoxalase1; HAB, high anxiety-related behavior; i.c.v., intracerebroventricular; LAB, low anxiety-related behavior; LTP, long-term potentiation; MG, methylglyoxal; NAB, normal anxiety-related behavior; TST, tail suspension test; WB, western blotting. AbstractMethylglyoxal (MG) is a highly reactive metabolite that forms adducts with basic amino acid side chains in proteins. MG is degraded by glyoxalase1 (GLO1), an enzyme shown to be differentially expressed in several mouse models of anxietyrelated behavior. As yet, molecular mechanisms by which altered GLO1 expression influences emotionality have not been elucidated. Here we report that both MG concentration and protein modification are altered in brain tissue of a mouse model for trait anxiety, with elevated levels in low anxiety-related behavior relative to high anxiety-related behavior animals. Accordingly, repeated intracerebroventricular injections of MG mediated anxiolysis in inbred high anxiety-related behavior and outbred CD1 mice. We found that anxiolytic-like properties of MG were independent of GLO1 expression. In contrast, antidepressant-like properties of intracerebroventricular MG were suppressed in CD1 mice carrying extra copies of the GLO1 gene. Moreover, MG treatment increased expression of GLO1 only in CD1 mice that did not have extra copies of GLO1. Taken together, these results suggest that the MG levels in brain are negatively correlated with anxiety. Thereby, we identified a novel molecular mec...

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“…A novel derivatization procedure was reported by Wu et al [104] to enable the simultaneous analysis of 3-DG and other dicarbonyls in human plasma, using two derivatization steps. Firstly with O-(2,3,4,5,6-pentafluorobenzyl) hydroxylamine (PFBOA; 2 h, 50 • C) and secondly with MSTFA (90 • C, 30 min) (Fig.…”

Section: Derivatives Of Intermediate Compoundsmentioning

confidence: 99%

Derivatization of carbohydrates for GC and GC–MS analyses

Ruiz‐Matute

Hernández‐Hernández

Rodríguez-Sánchez

et al. 2011

Journal of Chromatography B

366

195

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A novel derivatization approach for simultaneous determination of glyoxal, methylglyoxal, and 3-deoxyglucosone in plasma by gas chromatography–mass spectrometry

Cited by 29 publications

References 23 publications

Induction of a Stringent Metabolic Response in Intracellular Stages of Leishmania mexicana Leads to Increased Dependence on Mitochondrial Metabolism

Induction of a Stringent Metabolic Response in Intracellular Stages of Leishmania mexicana Leads to Increased Dependence on Mitochondrial Metabolism

Methylglyoxal‐mediated anxiolysis involves increased protein modification and elevated expression of glyoxalase 1 in the brain

Derivatization of carbohydrates for GC and GC–MS analyses

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