RESUMEN
The aim of the study was a systematic evaluation of cognitive development in individuals with glutaric aciduria type 1 (GA1), a rare neurometabolic disorder, identified by newborn screening in Germany. This national, prospective, observational, multi-centre study includes 107 individuals with confirmed GA1 identified by newborn screening between 1999 and 2020 in Germany. Clinical status, development, and IQ were assessed using standardized tests. Impact of interventional and non-interventional parameters on cognitive outcome was evaluated. The majority of tested individuals (n = 72) showed stable IQ values with age (n = 56 with IQ test; median test age 11 years) but a significantly lower performance (median [IQR] IQ 87 [78-98]) than in general population, particularly in individuals with a biochemical high excreter phenotype (84 [75-96]) compared to the low excreter group (98 [92-105]; p = 0.0164). For all patients, IQ results were homogenous on subscale levels. Sex, clinical motor phenotype and quality of metabolic treatment had no impact on cognitive functions. Long-term neurologic outcome in GA1 involves both motor and cognitive functions. The biochemical high excreter phenotype is the major risk factor for cognitive impairment while cognitive functions do not appear to be impacted by current therapy and striatal damage. These findings implicate the necessity of new treatment concepts.
Asunto(s)
Errores Innatos del Metabolismo de los Aminoácidos/complicaciones , Encefalopatías Metabólicas/complicaciones , Desarrollo Infantil , Disfunción Cognitiva/epidemiología , Glutaratos/orina , Glutaril-CoA Deshidrogenasa/deficiencia , Adolescente , Errores Innatos del Metabolismo de los Aminoácidos/diagnóstico , Errores Innatos del Metabolismo de los Aminoácidos/metabolismo , Errores Innatos del Metabolismo de los Aminoácidos/orina , Encefalopatías Metabólicas/diagnóstico , Encefalopatías Metabólicas/metabolismo , Encefalopatías Metabólicas/orina , Niño , Preescolar , Disfunción Cognitiva/diagnóstico , Disfunción Cognitiva/etiología , Disfunción Cognitiva/metabolismo , Femenino , Estudios de Seguimiento , Alemania/epidemiología , Glutaratos/metabolismo , Glutaril-CoA Deshidrogenasa/metabolismo , Glutaril-CoA Deshidrogenasa/orina , Humanos , Lactante , Recién Nacido , Pruebas de Inteligencia/estadística & datos numéricos , Masculino , Tamizaje Neonatal/métodos , Estudios Prospectivos , Medición de Riesgo/métodos , Adulto JovenRESUMEN
The paper reports on monitoring methylmalonic aciduria (MMA)-specific and non-specific metabolites via NMR urinomics. Five patients have been monitored over periods of time; things involved were diet, medication and occasional episodes of failing to comply with prescribed diets. An extended dataset of targeted metabolites is presented, and correlations with the type of MMA are underlined. A survey of previous NMR studies on MMA is also presented.
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Errores Innatos del Metabolismo de los Aminoácidos/orina , Espectroscopía de Resonancia Magnética , Metabolómica , Niño , Preescolar , Creatinina/orina , Femenino , Glicina/orina , Humanos , Lactante , Recién Nacido , Masculino , Factores de TiempoRESUMEN
BACKGROUND: Aromatic L-amino acid decarboxylase (AADC) deficiency is a primary neurotransmitter defect of the biosynthesis of catecholamines and serotonin. The phenotype consists of varying degrees of neurological impairment, including motor and non-motor symptoms. Treatment outcomes correlate with the time point of diagnosis and treatment initiation; therefore, reliable diagnostic markers are necessary. Increased vanillactic acid (VLA) concentrations in the analysis of organic acids in urine have been reported in AADC deficiency. However, this elevation is often subtle and easily missed. In this study, we evaluate the semi-quantitative determination of VLA and vanillylmandelic acid (VMA) concentrations and establish the ratio of a VLA/VMA as a novel diagnostic marker for AADC deficiency. METHODS: Urine samples obtained from 10,095 non-AADC deficient controls and 14 confirmed AADC deficient patients were used for organic acid analysis by liquid-liquid extraction of the acidified samples and gas chromatographic-mass spectrometric separation after trimethylsilylation. The semi-quantitative determination of VLA and VMA concentrations and the calculation of a VLA/VMA ratio were evaluated as a diagnostic marker for AADC deficiency. RESULTS: The mean VLA and VMA concentrations in 10,095 non-AADCD samples was 0.3 mmol/mol creatinine (SD = 1.18, range 0-57.79) and 5.59 mmol/mol creatinine (SD = 3.87, range 0.04-60.62), respectively. The mean concentration of VLA in 14 patient-derived samples was 10.24 mmol/mol creatinine, (SD = 11.58, range = 0.37-33.06) and 0.45 mmol/mol creatinine for VMA (SD = 0.29, range 0.11-1.27). The mean VLA/VMA ratio in non-AADC controls was 0.07 (SD = 0.37, range 0.0-23.24), whereas AADC deficient patients revealed a mean VLA/VMA ratio of 23.16 (SD = 22.83, range 0.97-74.1). The VLA/VMA ratio thus allows a reliable identification of patients with AADC deficiency, especially in the young age cohort as it decreases with age. To take this into account, age-adjusted thresholds have been developed. CONCLUSION: Determination of individual concentrations of VLA and VMA in urine does not allow a reliable diagnosis of AADC deficiency. In this study, we could demonstrate that a semi-quantitative analysis of organic acids in urine allows the formation of metabolite ratios and that the VLA/VMA ratio is a reliable, easily accessible, new parameter for the diagnosis of AADC deficiency.
Asunto(s)
Errores Innatos del Metabolismo de los Aminoácidos/genética , Errores Innatos del Metabolismo de los Aminoácidos/orina , Descarboxilasas de Aminoácido-L-Aromático/deficiencia , Descarboxilasas de Aminoácido-L-Aromático/genética , Biomarcadores/orina , Adolescente , Adulto , Errores Innatos del Metabolismo de los Aminoácidos/patología , Descarboxilasas de Aminoácido-L-Aromático/orina , Niño , Preescolar , Femenino , Ácido Homovanílico/análogos & derivados , Ácido Homovanílico/orina , Humanos , Lactante , Masculino , Ácido Vanilmandélico/orina , Adulto JovenRESUMEN
Mercaptopyruvate sulfurtransferase (Mpst) and its homolog thiosulfate sulfurtransferase (Tst = rhodanese) detoxify cyanide to thiocyanate. Mpst is attracting attention as one of the four endogenous hydrogen sulfide (H2S)/reactive sulfur species (RSS)-producing enzymes, along with cystathionine ß-synthase (Cbs), cystathionine γ-lyase (Cth), and cysteinyl-tRNA synthetase 2 (Cars2). MPST deficiency was found in 1960s among rare hereditary mercaptolactate-cysteine disulfiduria patients. Mpst-knockout (KO) mice with enhanced liver Tst expression were recently generated as its model; however, the physiological roles/significances of Mpst remain largely unknown. Here we generated three independent germ lines of Mpst-KO mice by CRISPR/Cas9 technology, all of which maintained normal hepatic Tst expression/activity. Mpst/Cth-double knockout (DKO) mice were generated via crossbreeding with our previously generated Cth-KO mice. Mpst-KO mice were born at the expected frequency and developed normally like Cth-KO mice, but displayed increased urinary 3-mercaptolactate excretion and enhanced passive systemic anaphylactic responses when compared to wild-type or Cth-KO mice. Mpst/Cth-DKO mice were also born at the expected frequency and developed normally, but excreted slightly more 3-mercaptolactate in urine compared to Mpst-KO or Cth-KO mice. Our Mpst-KO, Cth-KO, and Mpst/Cth-DKO mice, unlike semi-lethal Cbs-KO mice and lethal Cars2-KO mice, are useful tools for analyzing the unknown physiological roles of endogenous H2S/RSS production.
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Errores Innatos del Metabolismo de los Aminoácidos/etiología , Errores Innatos del Metabolismo de los Aminoácidos/metabolismo , Compuestos de Sulfhidrilo/orina , Sulfurtransferasas/deficiencia , Alelos , Errores Innatos del Metabolismo de los Aminoácidos/orina , Animales , Biomarcadores , Modelos Animales de Enfermedad , Marcación de Gen , Genotipo , Hígado/metabolismo , Ratones , Ratones Noqueados , MutaciónAsunto(s)
Acetil-CoA C-Aciltransferasa/deficiencia , Acidosis , Errores Innatos del Metabolismo de los Aminoácidos/diagnóstico , Cuerpos Cetónicos/sangre , Vómitos , Acetoacetatos/orina , Acetil-CoA C-Aciltransferasa/sangre , Acetil-CoA C-Aciltransferasa/orina , Acidosis/sangre , Acidosis/complicaciones , Acidosis/orina , Errores Innatos del Metabolismo de los Aminoácidos/sangre , Errores Innatos del Metabolismo de los Aminoácidos/orina , Glicina/análogos & derivados , Glicina/orina , Humanos , Hidroxibutiratos/orina , Lactante , Masculino , Vómitos/sangre , Vómitos/etiología , Vómitos/orinaRESUMEN
Objective: To improve the awareness of methylmalonic acidemia and hyperhomocysteinemia with diffuse lung disease as an initial or main presentation. Methods: A retrospective analysis of the clinical manifestations, radiological features, laboratory tests, genetic variations, treatments and prognoses was conducted in six children presented with diffuse lung disease and finally diagnosed with methylmalonic acidemia and hyperhomocysteinemia in Ward 2 of Department of Respiratory Diseases, Beijing Children's Hospital, from August 2017 to November 2018. Results: Six children were included in this study. Two children were male and four were female. The average age of onset was 28 months. The mean age at diagnosis was 34 months. The average interval from onset to diagnosis was 6 months. Four children who underwent genetic tests were found to have variants of gene MMACHC and diagnosed with CblC type. All children had respiratory symptoms and signs as initial or main presentation, which were tachypnea (5 cases), exercise intolerance (5 cases), cough (4 cases), cyanosis (4 cases), clubbing (4 cases), dyspnea (3 cases) and retractions (3 cases). Pulmonary arterial hypertension was found in all six children. Pericardial effusion (4 cases), kidney involvement (3 cases), nervous system involvement (3 cases), gastrointestinal system involvement (3 cases) and anemia (2 cases) also coexisted. The high resolution computed tomography (HRCT) features included dilated pulmonary artery (6 cases), ground-glass opacities (4 cases), diffuse poorly defined ground-glass centrilobular nodules (3 cases), pleural effusion (3 cases), thickening of interlobular septum (2 cases), etc. All children had an elevated concentration of methylmalonic acid in urine and homocysteine in plasma. Genetic tests were performed in four patients, and MMACHC genetic mutations were found in all of them. Clinical manifestations, HRCT features and pulmonary arterial hypertension turned better in five children after treatment. One patient who was not regularly followed-up died. Conclusions: Pulmonary involvement including diffuse lung disease and pulmonary arterial hypertension could coexist with methylmalonic acidemia and hyperhomocysteinemia, which may have respiratory symptoms and signs as the initial or main presentation. Characteristic HRCT features were found in some patients. Plasma homocysteine test is a quick method for screening the disease in children with diffuse lung disease and (or) pulmonary arterial hypertension. Both diffuse lung disease and pulmonary arterial hypertension may turn better after treatment.
Asunto(s)
Errores Innatos del Metabolismo de los Aminoácidos/diagnóstico , Hiperhomocisteinemia/diagnóstico , Enfermedades Pulmonares/etiología , Pulmón/fisiopatología , Arteria Pulmonar/diagnóstico por imagen , Edad de Inicio , Errores Innatos del Metabolismo de los Aminoácidos/orina , Proteínas Portadoras , Niño , Preescolar , Femenino , Humanos , Hiperhomocisteinemia/sangre , Lactante , Masculino , Oxidorreductasas , Estudios Retrospectivos , Tomografía Computarizada por Rayos XRESUMEN
Methyl malonic acid and branched-chain keto acids are important biomarkers for the diagnosis of cobalamin deficiencies and maple syrup urine disease. We report the development and validation of a HILIC-ESI-MS2 method for the quantification of these organic acids from neonatal urine. The samples were 100 times diluted and analyzed on a ZIC-HILIC column with 25-mM formic acid in water: 25-mM formic acid in acetonitrile (45:55) at a flow rate of 0.8 mL/min with a runtime of only 6 minutes. The method demonstrated a lower limit of detection of 10 ng/mL, Limit of Quantification (LOQ) of 50 ng/mL, linearity of r2 ≥ 0.990 and recoveries of 87-105% for all analytes. The intraday and interday precision CV's were <10% and 12%, respectively. Extensive stability studies demonstrated the analytes to be stable in stock and in matrix with a percent change within ±15%. The Bland-Altman analysis of the developed method with the gold standard GCMS method demonstrated a bias of 0.44, 0.11, 0.009 and -0.19 for methyl malonic acid, 3-methyl-2-oxovaleric acid, 2-hydroxy-3methylbutyric acid and 4-methyl-2-oxovaleric acid, respectively, proving the methods are comparable. The newly developed method involves no derivatization and has a simple sample preparation and a low runtime, enabling it to be easily automated with a high sample throughput in a cost-effective manner.
Asunto(s)
Errores Innatos del Metabolismo de los Aminoácidos/orina , Biomarcadores/orina , Cromatografía Líquida de Alta Presión/métodos , Enfermedad de la Orina de Jarabe de Arce/orina , Espectrometría de Masa por Ionización de Electrospray/métodos , Errores Innatos del Metabolismo de los Aminoácidos/diagnóstico , Humanos , Malonatos/orina , Enfermedad de la Orina de Jarabe de Arce/diagnósticoRESUMEN
3-hydroxy-3-methylglutaric aciduria (HMGA) is an inherited disorder of the leucine catabolic pathway in which occurs a deficiency of the 3-hydroxy-3-methylglutaryl-CoA lyase enzyme. Therefore, the organic acids 3-hydroxy-3-methylglutaric (HMG) and 3-methylglutaric (MGA), mainly, accumulate in tissues of affected patients. Lately, much attention has been focused on free radicals as mediators of tissue damage in human diseases, causing lipid peroxidation, protein oxidation and DNA damage. The treatment of this disease is based in a restricted protein ingest and supplementation with l-carnitine (LC), an antioxidant and detoxifying agent. In the present work, we investigated the in vitro oxidative damage to DNA induced by the accumulation of organic acids and oxidative stress parameters in vivo of patients with 3-HMG, as well as the effect of the recommended therapy. The in vitro DNA damage was analyzed by the alkaline comet assay in leukocytes incubated with HMG and MGA (1â¯mM, 2.5â¯mM and 5â¯mM) and co-incubated with LC (90⯵M and 150⯵M). The in vivo urinary 15-F2t-isoprostane levels and urinary oxidized guanine species were measured by ELISA kits in patient's urine before and after the treatment with LC. HMG and MGA induced a DNA damage index (DI) significantly higher than that of the control group. The DI was significantly reduced in the presence of LC. It was also verified a significant increase of oxidized guanine species and urinary isoprostane levels, biomarker of oxidative DNA damage and lipid peroxidation respectively, in patients before treatment. After the treatment and supplementation with LC, patients presented significantly lower levels of those biomarkers. Analyzing the data together, we can conclude that HMGA patients present oxidative lipid and DNA damage, which is induced by HMG and MGA, and the antioxidant therapy with LC can prevent that kind of injuries.
Asunto(s)
Acetil-CoA C-Acetiltransferasa/deficiencia , Errores Innatos del Metabolismo de los Aminoácidos/tratamiento farmacológico , Carnitina/uso terapéutico , Daño del ADN/efectos de los fármacos , Meglutol/análogos & derivados , Meglutol/metabolismo , 8-Hidroxi-2'-Desoxicoguanosina/orina , Acetil-CoA C-Acetiltransferasa/metabolismo , Acetil-CoA C-Acetiltransferasa/orina , Adolescente , Errores Innatos del Metabolismo de los Aminoácidos/metabolismo , Errores Innatos del Metabolismo de los Aminoácidos/orina , Niño , Preescolar , Dinoprost/análogos & derivados , Dinoprost/orina , Guanina/análogos & derivados , Guanina/orina , Guanosina/análogos & derivados , Guanosina/orina , Humanos , Lactante , Peroxidación de Lípido/efectos de los fármacosRESUMEN
A novel method utilizing ambient thermal desorption ionization with a direct analysis in real-time source integrated with mass spectrometry (DART-MS) was established and applied to the rapid analysis of 3-hydroxy-3-methylglutaric (3-HMG) acid in the neonatal urine. Instrument parameter settings were optimized to obtain high sensitive and accurate determination of 3-HMG acid. The use of helium gas heated to temperature of 400°C was observed to permit deprotonation, 3-HMG acid producing an abundant (M-H)- (m/z 161) in the negative ion mode. The calibration curve was determined to be linear over the range of 0.05-5 mg/L, with the correlation coefficient r = 0.9988 and the relative standard deviations (n = 6) in the range of 1.5-11.8%. The limit of detection was 0.002 mg/L, and the limit of quantitation was 0.007 mg/L. The recoveries ranged from 88.0% to 123.1%. Four urine samples from patients and four simulated urine samples were investigated. The results of DART-MS were in agreement with the values determined using established methods at the hospitals. The proposed method demonstrated significant potential in the application of the high-throughput screening in newborn screening.
Asunto(s)
Acetil-CoA C-Acetiltransferasa/deficiencia , Errores Innatos del Metabolismo de los Aminoácidos/diagnóstico , Acetil-CoA C-Acetiltransferasa/orina , Errores Innatos del Metabolismo de los Aminoácidos/orina , Humanos , Recién Nacido , Espectrometría de Masas/métodos , Tamizaje NeonatalRESUMEN
Accumulation of methylcitrate is a biochemical hallmark of inborn errors of propionate metabolism, a group of disorders that include propionic acidemia, methylmalonic aciduria and cobalamin defects. In clinical laboratories, this analyte is measured without quantification by gas chromatography mass spectrometry as part of urine organic acids. Here we describe a simple, sensitive and specific method to quantify methylcitrate in dried urine spots by liquid chromatography tandem mass spectrometry. Methylcitrate is extracted and derivatized with 4-[2-(N,N-dimethylamino)ethylaminosulfonyl]-7-(2-aminoethylamino)-2,1,3-benzoxadiazole in a single step. A derivatization mixture was added to 3.2â¯mm disc of dried urine spots, incubated at 65⯰C for 45â¯min and 4⯵l of the reaction mixture were analyzed. Separation was achieved on C18 column with methylcitrate eluting at 3.8â¯min. Intraday and interday imprecision (nâ¯=â¯17) were ≤20.9%. The method was applied on dried urine spots from established patients and controls. In controls (nâ¯=â¯135), methylcitrate reference interval of 0.4-3.4â¯mmol/mol creatinine. In patients, methylcitrate ranged between 8.3 and 591â¯mmol/mol creatinine. Quantification of methylcitrate provides important diagnostic clues for propionic acidemia, methylmalonic aciduria and cobalamin disorders. The potential utilization of methylcitrate as monitoring biomarker of patients under treatment and whether it correlates with the clinical status has yet to be established.
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Errores Innatos del Metabolismo de los Aminoácidos/diagnóstico , Citratos/orina , Acidemia Propiónica/diagnóstico , Urinálisis , Errores Innatos del Metabolismo de los Aminoácidos/orina , Cromatografía Liquida , Humanos , Acidemia Propiónica/orina , Espectrometría de Masas en TándemRESUMEN
AIM: Molybdenum co-factor deficiencies and isolated sulfite oxidase deficiency are rare autosomal recessively inherited diseases characterized by severe psychomotor impairment, intractable seizures, dislocated lens and dysmorphic facial features. The biochemical diagnosis of these diseases requires the determination of urine sulfocysteine. MATERIALS & METHODS: Urine sulfocysteine was quantified by an ultra-high performance liquid chromatography-MS/MS assay. The method was validated for linearity, accuracy, precision, recovery and stability. RESULTS & CONCLUSION: Total imprecision of accuracy was less than 6%. Intra-assay and inter-assay precisions were less than 5%. The recovery was higher than 98%. The method is inexpensive, fast, accurate and has been successfully used for identifying five molybdenum co-factor deficient and six sulfite oxidase deficient patients since deployed.
Asunto(s)
Errores Innatos del Metabolismo de los Aminoácidos/diagnóstico , Errores Innatos del Metabolismo de los Aminoácidos/orina , Cisteína/análogos & derivados , Molibdeno/deficiencia , Sulfito-Oxidasa/deficiencia , Urinálisis/métodos , Cromatografía Líquida de Alta Presión , Coenzimas/deficiencia , Cisteína/orina , Humanos , Límite de Detección , Modelos Lineales , Sulfito-Oxidasa/orina , Espectrometría de Masas en Tándem , Factores de TiempoAsunto(s)
Errores Innatos del Metabolismo de los Aminoácidos/diagnóstico , Errores Innatos del Metabolismo de los Aminoácidos/orina , Glutaratos/orina , Adolescente , Errores Innatos del Metabolismo de los Aminoácidos/diagnóstico por imagen , Encéfalo/diagnóstico por imagen , Niño , Discapacidades del Desarrollo/etiología , Humanos , Discapacidad Intelectual/etiología , Imagen por Resonancia Magnética , Megalencefalia/etiología , Hermanos , Temblor/etiologíaRESUMEN
OBJECTIVE: To determine the frequency of disorders leading to methylmalonic acidurias. METHODS: This cross-sectional study was conducted from January 2013 to April 2016 at the Aga Khan University Hospital, Karachi, and comprised patients diagnosed with methylmalonic acidurias based on urine organic acid analysis. Clinical history and biochemical data was collected from the biochemical genetics laboratory requisition forms. Organic acid chromatograms of all the subjects were critically reviewed by a biochemical pathologist and a metabolic physician. For assessing the clinical outcome, medical charts of the patients were reviewed. SPSS 19 was used for data analysis. RESULTS: Of the 1,778 patients 50(2.81%) were detected with methylmalonic acidurias. After excluding patients with non-significant peaks of methylmalonic acidemia, 41(2.31%) were included in the final analysis. Of these, 20(48.7%) were females, while the overall median age was 11.5 months (interquartile range: 6-41.5). On stratification by type of disorders leading to methylmalonic acidurias, 9(22%) had methylmalonic acidemia, 12(29%) had Cobalamin-related remethylation disorders, nonspecific methylmalonic acidurias in 16(39%), while 2(5%) each had succinyl coenzyme A synthetase and Vitamin B12 deficiency. respectively. CONCLUSIONS: Screening tests, including urine organic acid, provided valuable clues to the aetiology of methylmalonic acidurias.
Asunto(s)
Errores Innatos del Metabolismo de los Aminoácidos/diagnóstico , Errores Innatos del Metabolismo de los Aminoácidos/etiología , Enfermedades Mitocondriales/complicaciones , Deficiencia de Vitamina B 12/complicaciones , Alanina/sangre , Errores Innatos del Metabolismo de los Aminoácidos/sangre , Errores Innatos del Metabolismo de los Aminoácidos/orina , Preescolar , Citratos/orina , Estudios Transversales , Femenino , Glicina/análogos & derivados , Glicina/sangre , Glicina/orina , Humanos , Lactante , Ácido Láctico/análogos & derivados , Ácido Láctico/orina , Masculino , Metionina/sangre , Pakistán , Centros de Atención Terciaria , Urinálisis/métodos , Valeratos/orinaRESUMEN
Blue diaper syndrome (BDS) (Online Mendelian Inheritance in Man number 211000) is an extremely rare disorder that was first described in 1964. The characteristic finding is a bluish discoloration of urine spots in the diapers of affected infants. Additional clinical features of the first described patients included diarrhea, inadequate weight gain, hypercalcemia, and nephrocalcinosis. An intestinal defect of tryptophan absorption was postulated as the underlying pathology. However, functional evidence for this theory is lacking. No genetic cause has been identified so far. Here, we report on a boy who presented with neonatal-onset diarrhea, metabolic acidosis, transient hepatopathy, recurrent hypoglycemia, and blue-stained urine spots in his diapers. An ultra-performance liquid chromatography-electrospray ionization-tandem mass spectrometry analysis of urine samples at different time points demonstrated the constant presence of indigo derivatives, thereby confirming the diagnosis of BDS. Of note, the visibility of indigo derivatives in the urine was highly dependent on the urine's pH. To identify the underlying genetic cause of the disease, whole-exome sequencing was performed, leading to the identification of a homozygous frameshift mutation in proprotein convertase subtilisin/kexin type 1 (PCSK1; NM_000439.4: c.679del, p.[Val227Leufs*12]). PCSK1 encodes prohormone convertase 1/3, and mutations within this gene have been reported as a rare cause of early-onset malabsorptive diarrhea and multiple endocrine dysfunction. In our report, we suggest that BDS can be caused by PCSK1 mutations.
Asunto(s)
Errores Innatos del Metabolismo de los Aminoácidos/genética , Anomalías del Ojo/genética , Mutación del Sistema de Lectura , Hipercalcemia/genética , Enfermedades Renales/genética , Proproteína Convertasa 1/genética , Errores Innatos del Metabolismo de los Aminoácidos/diagnóstico , Errores Innatos del Metabolismo de los Aminoácidos/orina , Cromatografía por Intercambio Iónico , Anomalías del Ojo/diagnóstico , Anomalías del Ojo/orina , Humanos , Concentración de Iones de Hidrógeno , Hipercalcemia/diagnóstico , Hipercalcemia/orina , Recién Nacido , Enfermedades Renales/diagnóstico , Enfermedades Renales/orina , Masculino , Espectrometría de Masas en Tándem , Triptófano/genética , Triptófano/orina , Orina/química , Secuenciación del ExomaRESUMEN
Organic acidurias (synonym, organic acid disorders, OADs) are a heterogenous group of inherited metabolic diseases delineated with the implementation of gas chromatography/mass spectrometry in metabolic laboratories starting in the 1960s and 1970s. Biochemically, OADs are characterized by accumulation of mono-, di- and/or tricarboxylic acids ("organic acids") and corresponding coenzyme A, carnitine and/or glycine esters, some of which are considered toxic at high concentrations. Clinically, disease onset is variable, however, affected individuals may already present during the newborn period with life-threatening acute metabolic crises and acute multi-organ failure. Tandem mass spectrometry-based newborn screening programmes, in particular for isovaleric aciduria and glutaric aciduria type 1, have significantly reduced diagnostic delay. Dietary treatment with low protein intake or reduced intake of the precursor amino acid(s), carnitine supplementation, cofactor treatment (in responsive patients) and nonadsorbable antibiotics is commonly used for maintenance treatment. Emergency treatment options with high carbohydrate/glucose intake, pharmacological and extracorporeal detoxification of accumulating toxic metabolites for intensified therapy during threatening episodes exist. Diagnostic and therapeutic measures have improved survival and overall outcome in individuals with OADs. However, it has become increasingly evident that the manifestation of late disease complications cannot be reliably predicted and prevented. Conventional metabolic treatment often fails to prevent irreversible organ dysfunction with increasing age, even if patients are considered to be "metabolically stable". This has challenged our understanding of OADs and has elicited the discussion on optimized therapy, including (early) organ transplantation, and long-term care.
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Errores Innatos del Metabolismo de los Aminoácidos/diagnóstico , Errores Innatos del Metabolismo de los Aminoácidos/terapia , Adulto , Errores Innatos del Metabolismo de los Aminoácidos/metabolismo , Errores Innatos del Metabolismo de los Aminoácidos/mortalidad , Errores Innatos del Metabolismo de los Aminoácidos/orina , Encefalopatías Metabólicas/metabolismo , Encefalopatías Metabólicas/orina , Glutaril-CoA Deshidrogenasa/deficiencia , Glutaril-CoA Deshidrogenasa/metabolismo , Glutaril-CoA Deshidrogenasa/orina , Humanos , Recién Nacido , Ácido Metilmalónico/metabolismo , Tamizaje Neonatal , Valor Predictivo de las Pruebas , Propionatos/metabolismo , Espectrometría de Masas en TándemRESUMEN
Methylmalonyl-CoA epimerase (MCE) converts d-methylmalonyl-CoA epimer to l-methylmalonyl-CoA epimer in the propionyl-CoA to succinyl-CoA pathway. Only seven cases of MCE deficiency have been described. In two cases, MCE deficiency was combined with sepiapterin reductase deficiency. The reported clinical pictures of isolated MCE are variable, with two asymptomatic patients and two other patients presenting with metabolic acidosis attacks. For combined MCE and sepiapterin reductase deficiency, the clinical picture is dominated by neurologic alterations. We report isolated MCE deficiency in a boy who presented at five years of age with acute metabolic acidosis. Metabolic investigations were consistent with propionic aciduria (PA). Unexpectedly, propionyl-CoA carboxylase activity was within the reference range. Afterward, apparently intermittent and mild excretion of methylmalonic acid (MMA) was discovered. Methylmalonic pathway gene set analysis using the next-generation sequencing approach allowed identification of the common homozygous nonsense pathogenic variant (c.139C > T-p.Arg47*) in the methylmalonyl-CoA epimerase gene (MCEE). Additional cases of MCE deficiency may help provide better insight regarding the clinical impact of this rare condition. MCE deficiency could be considered a cause of mild and intermittent increases in methylmalonic acid.
Asunto(s)
Errores Innatos del Metabolismo de los Aminoácidos/diagnóstico , Acidemia Propiónica/diagnóstico , Racemasas y Epimerasas/deficiencia , Errores Innatos del Metabolismo de los Aminoácidos/genética , Errores Innatos del Metabolismo de los Aminoácidos/metabolismo , Errores Innatos del Metabolismo de los Aminoácidos/orina , Preescolar , Codón sin Sentido , Humanos , Masculino , Ácido Metilmalónico/metabolismo , Ácido Metilmalónico/orina , Racemasas y Epimerasas/genética , Racemasas y Epimerasas/metabolismo , Racemasas y Epimerasas/orinaRESUMEN
Tandem MS acylcarnitine "profiles" are extremely valuable. Although used appropriately in newborn screening programs to identify patients with possible diseases, their inadequate quantitative accuracy and lack of selectivity is problematic for confirmatory testing. In this report, we show the application of our validated, selective, accurate, precise, and robust UHPLC-MS/MS method for quantitation of acylcarnitines, specifically to C5 acylcarnitines: pivaloyl-, 2-methylbutyryl-, isovaleryl-, and valerylcarnitine. Standardized calibrants were used to generate 13-point, 200-fold concentration range calibration curves. Samples were isolated by solid-phase extraction and derivatized with pentafluorophenacyl trifluoromethanesulfonate. Acylcarnitine pentafluorophenacyl esters were eluted in 14min chromatograms. Data demonstrating quantitative stability and method robustness over a five year time period are shown and these results validate the method's accuracy and robustness. Urine from patients with isovaleric acidemia (with the disease marker isovalerylcarnitine) and with pivaloylcarnitine present are shown. These results demonstrate the method's ability to distinguish true isovaleric acidemia from pivalate derived interference. Our method for acylcarnitine quantitation is shown to be accurate, precise, and robust for selective quantitation of isovalerylcarnitine, and thus is recommended for confirmatory testing of suspected isovaleric acidemia patients.
Asunto(s)
Errores Innatos del Metabolismo de los Aminoácidos/orina , Carnitina/análogos & derivados , Cromatografía Líquida de Alta Presión/métodos , Isovaleril-CoA Deshidrogenasa/deficiencia , Espectrometría de Masas en Tándem/métodos , Carnitina/orina , Humanos , Isovaleril-CoA Deshidrogenasa/orina , Límite de Detección , Reproducibilidad de los ResultadosAsunto(s)
Distonía/complicaciones , Microcefalia/complicaciones , Errores Innatos del Metabolismo de los Aminoácidos/metabolismo , Errores Innatos del Metabolismo de los Aminoácidos/orina , Encefalopatías Metabólicas/metabolismo , Encefalopatías Metabólicas/orina , Distonía/diagnóstico , Distonía/orina , Cromatografía de Gases y Espectrometría de Masas , Glutaril-CoA Deshidrogenasa/deficiencia , Glutaril-CoA Deshidrogenasa/metabolismo , Glutaril-CoA Deshidrogenasa/orina , Humanos , Lactante , Imagen por Resonancia Magnética , Masculino , Microcefalia/diagnóstico , Microcefalia/orinaRESUMEN
BACKGROUND: 3-Hydroxy-3-Methylglutaryl-Coenzyme A (HMG-CoA) lyase deficiency is a rare inborn error of leucine metabolism and ketogenesis. Despite recurrent hypoglycemia and metabolic decompensations, most patients have a good clinical and neurological outcome contrasting with abnormal brain magnetic resonance imaging (MRI) signals and consistent abnormal brain proton magnetic resonance spectroscopy (1H-MRS) metabolite peaks. Identifying these metabolites could provide surrogate markers of the disease and improve understanding of MRI-clinical discrepancy and follow-up of affected patients. METHODS: Urine samples, brain MRI and 1H-MRS in 5 patients with HMG-CoA lyase deficiency (4 boys and 1 girl aged from 25days to 10years) were, for each patient, obtained on the same day. Brain and urine spectroscopy were performed at the same pH by studying urine at pH 7.4. Due to pH-induced modifications in chemical shifts and because reference 1H NMR spectra are obtained at pH 2.5, spectroscopy of normal urine added with the suspected metabolite was further performed at this pH to validate the correct identification of compounds. RESULTS: Mild to extended abnormal white matter MRI signals were observed in all cases. Brain spectroscopy abnormal peaks at 0.8-1.1ppm, 1.2-1.4ppm and 2.4ppm were also detected by urine spectroscopy at pH 7.4. Taking into account pH-induced changes in chemical shifts, brain abnormal peaks in patients were formally identified to be those of 3-hydroxyisovaleric, 3-methylglutaconic, 3-methylglutaric and 3-hydroxy-3-methylglutaric acids. CONCLUSION: 3-Methylglutaric, 3-hydroxyisovaleric and 3-hydroxy-3-methylglutaric acids identified on urine 1H-NMR spectra of 5 patients with HMG-CoA lyase deficiency are responsible for the cerebral spectroscopy signature seen in these patients, validating their local involvement in brain and putative contribution to brain neuropathology.
Asunto(s)
Acetil-CoA C-Acetiltransferasa/deficiencia , Errores Innatos del Metabolismo de los Aminoácidos/metabolismo , Errores Innatos del Metabolismo de los Aminoácidos/orina , Química Encefálica , Encéfalo/diagnóstico por imagen , Encéfalo/metabolismo , Meglutol/orina , Metabolómica/métodos , Acetil-CoA C-Acetiltransferasa/química , Acetil-CoA C-Acetiltransferasa/metabolismo , Acetil-CoA C-Acetiltransferasa/orina , Errores Innatos del Metabolismo de los Aminoácidos/diagnóstico por imagen , Cerebelo/metabolismo , Niño , Preescolar , Femenino , Humanos , Concentración de Iones de Hidrógeno , Lactante , Recién Nacido , Imagen por Resonancia Magnética , Masculino , Meglutol/análogos & derivados , Meglutol/metabolismo , Espectroscopía de Protones por Resonancia Magnética , Orina/química , Valeratos/metabolismo , Sustancia Blanca/metabolismoRESUMEN
A new, precise, and very selective method for increasing the impact and assessment of histidine as a biomarker for early diagnosis of histidinemia disease in new born children was developed. The method depends on the formation of the ion pair associate between histidine and the nano optical samarium tetracycline [Sm-(TC)2]+ complex doped in sol-gel matrix in a borate buffer of pH 9.2. The [Sm-(TC)2]+ complex has +I net charge which is very selective and sensitive for [histidine]- at pH 9.2 in serum and urine samples of histidinemia disease. Histidine enhances the luminescence intensity of the nano optical [Sm-(TC)2]+ complex at 645nm after excitation at 400nm, in borate buffer, pH 9.2. The remarkable enhancement of the luminescence intensity at 645nm of nano [Sm-(TC)2]+ complex doped in sol-gel matrix by various concentrations of the histidine was successfully used as an optical probe for the assessment of histidine in different serum and urine samples of new born children infected by histidinemia. The calibration plot was achieved over the concentration range 1.4×10-5 - 6.5×10-10molL-1 histidine with a correlation coefficient of (0.998) and a detection limit of (3.2×10-10molL-1). The sensitivity (98.88%) and specificity (97.41%) of histidine as a biomarker were calculated.