Differentiation of long-chain fatty acid oxidation disorders using alternative precursors and acylcarnitine profiling in fibroblasts

Roe, Diane S.; Yang, Bing-Zhi; Vianey‐Saban, Christine; Struys, E.A; Sweetman, Lawrence; Roe, Charles R.

doi:10.1016/j.ymgme.2005.09.018

Cited by 27 publications

(19 citation statements)

References 22 publications

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“…Serum creatine phosphokinase (CPK) levels and blood acylcarnitine analyses were only abnormal during episodes of rhabdomyolysis. Only enzyme assay 4 and in vitro fibroblast analysis 6 were consistently reliable for diagnosis (see e-Patients and e-Methods).…”

Section: Patient Descriptionsmentioning

confidence: 99%

“…CPT II enzyme assays, 4 in vitro palmitate oxidation, 5,6 DNA analysis, 7,8 acylcarnitine analysis, 3 plasma fatty acid metabolites (ketone bodies), 9 and protein measurements 10 were performed for diagnostic and monitoring purposes. The 36-item Short-Form Health Status survey (SF-36) version 2, 11 bioelectric impedance measurements, and serial blood and urine chemical measurements were obtained on each patient (for more detail, see e-Methods on the Neurology ® Web site at www.neurology.org).…”

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confidence: 99%

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Carnitine palmitoyltransferase II deficiency

et al. 2008

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Section: Patient Descriptionsmentioning

confidence: 99%

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confidence: 99%

Carnitine palmitoyltransferase II deficiency

et al. 2008

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“…As a critical assay step, in situ permeabilization of cell membranes with digitonin allowed the study of an intact organelle environment in a much shorter incubation and assay turnaround time compared with wholecell incubation assays (12)(13)(14)(15)(16) ). An assay scheme including the analyzed metabolic pathway is provided in Fig.…”

Section: Resultsmentioning

confidence: 99%

“…In comparison, the amount of protein required for comparable whole-cell incubation systems is at least twice to three times as high ( 13,14,16 ). Thus, the method is applicable even when availability of cell material is sparse, e.g., in the investigation of primary cells such as human adipocytes.…”

Section: Resultsmentioning

confidence: 99%

“…[U- 14 C] hexadecanoate was used to measure acylcarnitine and acyl-CoA intermediates by radio-HPLC in mitochondrial preparations ( 9 ) and permeabilized human fi broblasts or peripheral blood cells ( 10,11 ). The development of MS/MS technology allowed the sensitive and specifi c quantitative profi ling of accumulating acylcarnitines following 72 or 96 h incubation of intact cells with stable isotopes or unlabeled substrates (12)(13)(14)(15)(16). A faster approach of MS/MS-based acylcarnitine profi ling was reported, applying incubation with stable isotopically labeled palm itate to cell homogenates containing intact mitochondria ( 17 ).…”

Section: Ms/ms Sample Preparationmentioning

confidence: 99%

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In situ assay of fatty acid β-oxidation by metabolite profiling following permeabilization of cell membranes

Ensenauer

Fingerhut

Schriever

et al. 2012

Journal of Lipid Research

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The mitochondrial FA ␤ -oxidation (FAO) pathway is a major contributor to cellular energy production and homeostasis, particularly once glycogen stores are depleted owing to fasting ( 1 ). Substantial amounts of energy are provided by FAO for heart, skeletal muscle, and liver function. In monogenetic defi ciencies of FAO or the carnitine shuttle system, accumulation of acyl-CoA intermediates and depletion of energy and free carnitine occur, specifically at times of increased energy demands, including fever and impaired food supply ( 2 ). Altered FA catabolism is considered key in the development of obesity-induced insulin resistance, because cellular overload with long-chain FAs is thought to exert a disastrous effect on mitochondrial function ( 3, 4 ). Cytoprotective mechanisms against FA lipotoxicity include channelling of FAs into triglyceride pools and enhancement of FA degradation via mitochondrial ␤ -oxidation ( 4 ).Thus, quantitative analysis of FAO has become increasingly important in assessing lipid-induced metabolic impairment and evaluating potential pharmacologic strategies designed to improve ␤ -oxidation fl ux ( 4 ). The most commonly applied method to assess functional FAO in cellular models has been the analysis of total FAO fl ux following incubation of intact cells with 14 C-or 3 H-labeled

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Metabolomic Responses of Human Hepatocytes to Emodin, Aristolochic Acid, and Triptolide: Chemicals Purified from Traditional Chinese Medicines

Liu

Cheng

et al. 2015

J Biochem Mol Toxicol

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The present study was undertaken to investigate the metabolic responses of human liver cells HL-7702 on chemicals purified from traditional Chinese medicine: emodin, triptolide, and aristolochic acid. Cytotoxicity tests demonstrated a dose-dependent toxic effect of emodin, triptolide, and aristolochic acid on HL7702 cells for 48 h. Emodin (14 μM), aristolochic acid (12 μg/mL), or triptolide (18 nM) was individually administrated to HL7702 and cell samples were collected after 48 h for metabolites extraction and analysis. Pattern recognition analysis reflected the significant difference in metabolic profiles between chemical-treated groups and the control group. Finally, eight metabolites including N1 -acetylspermidine, Glu Gly, N-undecanoylglycine, C16 sphinganine, sphinganine, glutathione, l-palmitoylcarnitine, and elaidic carnitine were detected as potential common biomarkers. Three pathways including sphinganine metabolism, fatty acid oxidation, and oxidative stress were identified. Our findings indicated that metabolomics would be an efficient approach to understand the molecular mechanism of hepatotoxicity induced by chemicals.

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Differentiation of long-chain fatty acid oxidation disorders using alternative precursors and acylcarnitine profiling in fibroblasts

Cited by 27 publications

References 22 publications

Carnitine palmitoyltransferase II deficiency

Carnitine palmitoyltransferase II deficiency

In situ assay of fatty acid β-oxidation by metabolite profiling following permeabilization of cell membranes

Metabolomic Responses of Human Hepatocytes to Emodin, Aristolochic Acid, and Triptolide: Chemicals Purified from Traditional Chinese Medicines

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