Blood polyunsaturated fatty acids in patients with peroxisomal disorders. A multicenter study (original) (raw)
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Plasma very long chain fatty acids in 3,000 peroxisome disease patients and 29,000 controls
Annals of Neurology, 1999
The assay of plasma very long chain fatty acids (VLCFAs), developed in our laboratory in 1981, has become the most widely used procedure for the diagnosis of X-linked adrenoleukodystrophy (X-ALD) and other peroxisomal disorders. We present here our 17 years' experience with this assay. Three VLCFA parameters, the level of hexacosanoic acid (C26:0), the ratio of C26:0 to tetracosanoic acid (C24:0), and of C26:0 to docosanoic acid (C22:0), were measured in 1,097 males (hemizygotes) with X-ALD, 1,282 women heterozygous for this disorder, including 379 obligate heterozygotes, 797 patients with other peroxisomal disorders, and 29,600 control subjects. All X-ALD hemizygotes who had not previously received Lorenzo's oil or a diet with a high erucic acid content had increased VLCFA levels, but the application of a discriminant function based on all three measurements is required to avoid the serious consequences of a false-negative result. VLCFA levels are increased at day of birth, thus providing the potential for neonatal mass screening, are identical in the childhood and adult forms, and do not change with age. Eighty-five percent of obligate heterozygotes had abnormally high VLCFA levels, but a normal result does not exclude carrier status. VLCFA levels were increased in all patients homozygous for Zellweger syndrome, neonatal adrenoleukodystrophy, infantile Refsum's disease, and in patients with deficiencies of peroxisomal acyl-coenzyme A oxidase, bifunctional enzyme, and 3-oxoacyl-coenzyme A thiolase. In these patients the degree of VLCFA excess correlated with clinical severity.
Folia Neuropathologica, 2009
Peroxisomal disorders are a large group of genetically determined metabolic diseases in which the biogenesis of peroxisomes is defective or there is a deficiency of only a single enzyme activity or substrate transporter. The objective of this report is to present ten years of experience in the diagnostics of peroxisomal disorders in Poland. Very-long-chain fatty acid (VLCFA) levels as a biomarker for peroxisomal defects were determined by gas chromatography in 1264 subjects with suspicion of peroxisome disease. Peroxisome biogenesis disorders (PBD) were diagnosed in 8 patients, bifunctional protein deficiency in 3 and X-linked adrenoleukodystrophy (X-ALD/AMN) in 127 hemi-or heterozygotes. The frequency of PBD was estimated as 0.20 : 100 000, and that of X-ALD/AMN 2.9 : 100 000 in Poland. Mean total delay time (onset of symptoms and diagnosis) for X-ALD/AMN was 2.2 years (range 0.25-13). High correlation of serum C26:0 concentration and survival for PBD patient (r 2 = 0.822; p < 0.001) was found.
Screening For Very Long Chain Fatty Acids in Egyptian Children with Inherited Peroxisomal Disorders
2015
Human peroxisomal disorders results from peroxisomal biogenesis disorders or from deficiency of a single peroxisomal enzyme or transporter. Peroxisomal disorders lead to a neurologic dysfunction of varying extent for most cases. The goal of this work is to establish a rapid procedure for the diagnosis of peroxisomal disorders for the quantification of very long chain fatty acids in plasma in addition to dried blood spot for comparison to evaluate a reliable, rapid method versus the cost. 50 patients clinically suspicious to have peroxisomal disorders and 25 normal controls were included in this study. Their ages ranged from 2 to 10 years. The patients include 45 (90%) males and 5 (10%) females. Positive consanguinity was present in 32 (64%) cases. Alanine aminotransferase and L-γ-Glutamyl transferase were measured for all patients to be sure that they are suffering from liver dysfunction. Very long chain fatty acids quantification in plasma and dried blood spots samples using labeled internal standard was done. The same analysis for the plasma samples was repeated using external unlabeled standards. Phytanic and pristanic acids were quantified. The results for alanine aminotransferase and L-γ-glutamyl transferase revealed their elevation in all patients compared to controls. The plasma and blood spot analysis using labeled internal standards showed that 6 patients (12%) from 50 had high C24:0, C26:0, C24:0/C22:0 and C26:0/C22:0, which prove the X-linked Adrenoleukodystrophy disorder. The plasma analysis without labeled internal standard showed that the same 6 patients had high C24:0, C26:0, C24:0/C22:0 and C26:0/C22:0, but the results were lower than those of the labeled internal standards except C24:0/C22:0 which was slightly higher. The phytanic acid and pristanic acid results showed a non-significant change. In conclusion, although using Electrospray Ionization Tandem Mass Spectrometry instrument with deuterated internal standards for the determination of very long chain fatty acids is an effective, precise, reliable and rapid screening procedure for many peroxisomal disorders in plasma samples and dried blood spots, using dried blood spots decrease the expensive collection and shipping of plasma samples. Measuring very long chain fatty acids without labeled internal standards decreased the cost, but it is not recommended due to absence of concentration correction effect of the labeled internal standards. In addition, using High Performance Liquid Chromatography with Electrospray Ionization Tandem Mass Spectrometry instrument for measuring phytanic and pristanic acids is a sensitive method, but needs double sample volume and more time for refining.
Metabolic Brain Disease, 2008
X-linked adrenoleukodystrophy (X-ALD) is an inherited disorder of peroxisomal metabolism, biochemically characterized by deficient β-oxidation of saturated very long chain fatty acids (VLCFA). The consequent accumulation of these fatty acids in different tissues and in biological fluids is associated with a progressive central and peripheral demyelination, as well as with adrenocortical insufficiency and hypogonadism. Seven variants of this disease have been described, being cerebral childhood the most frequent. The recommended therapy consists of the use of the glyceroltrioleate/glyceroltrierucate mixture known as Lorenzo’s Oil (LO), combined with a VLCFA-poor diet, but only in asymptomatic patients will this treatment prevent the progression of the symptomatology. In the present study we evaluated the biochemical course of patients with cerebral childhood (CCER) and asymptomatic clinical forms of X-ALD treated with LO associated with a VLCFA-restricted diet. We observed that hexacosanoic acid plasma concentrations and hexacosanoic/docosanoic ratio were significantly reduced in CCER patients during treatment when compared with diagnosis. Hexacosanoic acid plasma level was significantly reduced when compared with that at diagnosis and achieved the normal levels only in asymptomatic patients under LO treatment. In asymptomatic patients the magnitude of hexacosanoic acid decrease was higher than that of the CCER patients. These results show the good biochemical response of LO treatment in asymptomatic X-ALD patients. It is possible to suppose that this could be correlated with the prevention of the appearance of neurological signals in this group of patients treated with LO.
Biochemical Journal, 1990
The metabolism of [1-14C]lignoceric acid (C24:0) and [1-14C]tetracosatetraenoic acid (C24:4, n-6) was studied in normal skin fibroblast cultures and in cultures from patients with defects in peroxisomal β-oxidation (but normal peroxisomal numbers). Cells from X-linked adrenoleukodystrophy (ALD) patients with a presumed defect in a peroxisomal acyl-CoA synthetase, specific for fatty acids of carbon chain lengths greater than 22 (very-long-chain fatty acids; VLCFA), showed a relatively normal production of radiolabelled CO2 and water-soluble metabolites from [1-14C]C24:0. However, the products of synthesis from acetate de novo (released by β-oxidation), i.e. C16 and C18 fatty acids, were decreased, and carbon chain elongation of the fatty acid was increased. In contrast, cell lines from two patients with an unidentified lesion in peroxisomal β-oxidation (peroxisomal disease, PD) showed a marked deficiency in CO2 and water-soluble metabolite production, a decreased synthesis of C16 and...
Peroxisomal Fatty Acid β-Oxidation in Relation to Adrenoleukodystrophy
Developmental Neuroscience, 1991
The peroxisomal oxidation of the long chain fatty acid palmitate (C16:0) and the very long chain fatty acids lignocerate (C24:0) and cerotate (C26:0) was studied in freshly prepared homogenates of cultured skin fibroblasts from control individuals and patients with peroxisomal disorders. The peroxisomal oxidation of the fatty acids is almost completely dependent on the addition of ATP, coenzyme A (CoA), Mg2+ and NAD'. However, the dependency of the oxidation of palmitate on the concentration of the cofactors differs markedly from that of the oxidation of lignocerate and cerotate. The peroxisomal oxidation of all three fatty acid substrates is markedly deficient in fibroblasts from patients with the Zellweger syndrome, the neonatal form of adrenoleukodystrophy and the infantile form of Refsum disease, in accordance with the deficiency of peroxisomes in these patients. In fibroblasts from patients with X-linked adrenoleukodystrophy the peroxisomal oxidation of lignocerate and cerotate is impaired, but not that of palmitate. Competition experiments indicate that in fibroblasts, as in rat liver, distinct enzyme systems are responsible for the oxidation of palmitate on the one hand and lignocerate and cerotate on the other hand. Fractionation studies indicate that in rat liver activation of cerotate and lignocerate to cerotoyl-CoA and lignoceroyl-CoA, respectively, occurs in two subcellular fractions, the endoplasmic reticulum and the peroxisomes but not in the mitochondria. In homogenates of fibroblasts from patients lacking peroxisomes there is a small (25%) but significant deficiency of the ability to activate very long chain fatty acids. This deficient activity of very long chain fatty acyl-CoA synthetase is also observed in fibroblast homogenates from patients with X-linked adrenoleukodystrophy. We conclude that X-linked adrenoleukodystrophy is caused by a deficiency of peroxisomal very long chain fatty acyl-CoA synthetase.
The Journal of Lipid Research, 2008
Quantification of pristanic acid, phytanic acid, and very long chain fatty acids (i.e., hexacosanoic, tetracosanoic, and docosanoic acids) in plasma is the primary method for investigateing a multitude of peroxisomal disorders (PDs). Typically based on GC-MS, existing methods are time-consuming and laborious. In this paper, we present a rapid and specific liquid chromatography tandem mass spectrometric method based on derivatization with 4-[2-(N,N-dimethylamino)ethylaminosulfonyl]-7-(2-aminoethylamino)-2,1,3-benzoxadiazole (DAABD-AE). Derivatization was undertaken to improve the poor mass spectrometric properties of these fatty acids. Analytes in plasma (20 ml) were hydrolyzed, extracted, and derivatized with DAABD-AE in ?2 h. Derivatives were separated on a reverse-phase column and detected by positive-ion electrospray ionization tandem mass spectrometry with a 5 min injection-to-injection time. Calibration plots were linear over ranges that cover physiological and pathological concentrations. Intraday (n 5 12) and interday (n 5 10) variations at low and high concentrations were less than 9.2%. Reference intervals in normal plasma (n 5 250) were established for each compound and were in agreement with the literature. Using specimens from patients with established diagnosis (n 5 20), various PDs were reliably detected. In conclusion, this method allows for the detection of at least nine PDs in a 5 min analytical run. Furthermore, this derivatization approach is potentially applicable to other disease markers carrying the carboxylic group. Rapid UPLC-MS/MS method for routine analysis of plasma pristanic, phytanic, and very long chain fatty acid markers of peroxisomal disorders. J. Lipid Res.