Analysis of a child with carnitine palmitoyl transferase 1A deficiency due to variant of CPT1A gene / 中华医学遗传学杂志

OBJECTIVE@#To report on the clinical, metabolic and genetic characteristics of a child with carnitine palmitoyl transferase 1A (CPT1A) deficiency.@*METHODS@#Clinical data and the level of acylcarnitine for a child who initially presented as epilepsy were analyzed. Genomic DNA was extracted from peripheral blood samples of the child and her parents and subjected to next-generation sequencing (NGS).@*RESULTS@#Mass spectrometry of blood acylcarnitine indicated increased carnitine 0 (C0) and significantly increased C0/ (C16+C18). DNA sequencing revealed that the child has carried compound heterozygous variants of the CPT1A gene, namely c.1846G>A and c.2201T>C, which were respectively inherited from her mother and father.@*CONCLUSION@#CPT1A presenting initially as epilepsy was unreported previously. Analysis of blood acylcarnitine C0 and C0/ (C16 + C18) ratio and NGS are necessary for the identification and diagnosis of CPT1A deficiency. The c.1846G>A and c.2201T>C variants of the CPT1A gene probably underlay the disease in this child. Above finding has also enriched the spectrum of CPT1A gene variants.

Reference values of amino acids, acylcarnitines and succinylacetone by tandem mass spectrometry for use in newborn screening in southwest Colombia / Valores de referencia de aminoácidos, acilcarnitinas y succinilacetona por espectrometría de masas en tándem para su uso en el tamizaje neonatal en el suroccidente de Colombia

Abstract Introduction: Inborn errors of metabolism (IEM) represent an important public health problem due to current diagnosis and treatment limitations, poor life quality of affected patients, and consequent untimely child death. In contrast to classical methods, tandem mass spectrometry (MS/MS) has allowed simultaneous evaluation of multiple metabolites associated with IEM offering higher sensitivity, low false positive rates and high throughput. Aims: Determine concentration levels for amino acids and acylcarnitines in blood of newborns from Colombia, to establish reference values for further use in diagnosis of IEM. Methods: Implementation of a method to determine amino acids, acylcarnitines and succinylacetone in newborn dried blood spots using MS/MS, and its application in a cross-sectional study conducted in 891 healthy neonates from Cali and Quibdo cities is described. Results: fifty-seven analytes that allow the diagnosis of more than 40 different pathologies were tested. The method showed to be linear, precise and accurate. Healthy neonates 1-18 days of age were included, 523 from Cali and 368 from Quibdo; 52% male and 48% female. Age-related differences on the concentration levels of amino acids and acylcarnitines were observed whereas no significant differences by gender were found. Conclusion: The study has contributed to reveal the usual concentration levels of amino acids, acylcarnitines and succinylacetone that could be used as reference for the establishment of a newborn metabolic screening program in Colombia.

Resumen Introducción: Los Errores Innatos del metabolismo (EIM) representan un importante problema de salud pública debido a limitaciones en el tratamiento y diagnóstico oportuno, la pobre calidad de vida de los pacientes afectados, así como la muerte infantil prematura. Comparada con los métodos clásicos, la espectrometría de masas en tándem (MS/MS) ha permitido la evaluación simultánea de múltiples metabolitos asociados con EIM, con una alta sensibilidad, baja proporción de falsos positivos y alto rendimiento. Objetivos: Determinar los niveles de concentración de aminoácidos y acilcarnitinas en sangre de recién nacidos de Colombia, para establecer los valores normales para usarlos como referencia en el diagnóstico de EIM. Métodos: Aquí, se describe la implementación de un método para determinar aminoácidos, acilcarnitinas y succinilacetona en gotas de sangre seca de recién nacidos usando MS/MS, y su aplicación en un estudio de corte transversal realizado en 891 neonatos sanos de las ciudades de Cali y Quibdó. Resultados: Se evaluaron 57 analitos que permiten el diagnóstico de más de 40 patologías diferentes. El método mostró ser lineal, preciso y exacto. Se incluyeron neonatos sanos de 1-18 días de edad, 523 de Cali y 368 de Quibdó, 52% hombres y 48% mujeres. Se observaron diferencias en los niveles de concentración de aminoácidos y acilcarnitinas relacionadas con la edad, mientras que no se encontraron diferencias significativas por sexo. Conclusión: El estudio ha contribuido a revelar los niveles usuales de concentración de aminoácidos, acilcarnitinas y succinilacetona que pueden ser usados como referencia para el establecimiento del programa de tamizaje neonatal metabólico en Colombia.

Analysis of acylcarnitine profiles in umbilical cord blood and during the early neonatal period by electrospray ionization tandem mass spectrometry

Acylcarnitine profiling by electrospray ionization tandem mass spectrometry (ESI-MS/MS) is a potent tool for the diagnosis and screening of fatty acid oxidation and organic acid disorders. Few studies have analyzed free carnitine and acylcarnitines in dried blood spots (DBS) of umbilical cord blood (CB) and the postnatal changes in the concentrations of these analytes. We have investigated these metabolites in healthy exclusively breastfed neonates and examined possible effects of birth weight and gestational age. DBS of CB were collected from 162 adequate for gestational age neonates. Paired DBS of heel-prick blood were collected 4-8 days after birth from 106 of these neonates, the majority exclusively breastfed. Methanol extracts of DBS with deuterium-labeled internal standards were derivatized before analysis by ESI-MS/MS. Most of the analytes were measured using a full-scan method. The levels of the major long-chain acylcarnitines, palmitoylcarnitine, stearoylcarnitine, and oleoylcarnitine, increased by 27, 12, and 109%, respectively, in the first week of life. Free carnitine and acetylcarnitine had a modest increase: 8 and 11%, respectively. Propionylcarnitine presented a different behavior, decreasing 9% during the period. The correlations between birth weight or gestational age and the concentrations of the analytes in DBS were weak (r £ 0.20) or nonsignificant. Adaptation to breast milk as the sole source of nutrients can explain the increase of these metabolites along the early neonatal period. Acylcarnitine profiling in CB should have a role in the early detection of metabolic disorders in high-risk neonates.

Plasma carnitine in children with idiopathic epilepsy treated with old and new antiepileptic drugs

Prolonged antiepileptic drugs treatment can result in secondary carnitine deficiency. Clinical studies indicate a decrease in free and total carnitine in children treated with old-generation antiepileptic drugs [especially valproate]. Some studies on valproic acid [VPA]-induced hepatotoxicity showed decreased free serum carnitine, but some did not. A number of studies on the effect of VPA and/or other antiepileptic drugs on carnitine concentrations yielded contradictory results. The effect of new antiepileptic drugs as oxcarbazepine and lamotrigine on carnitine metabolism has not been reported previously. The aim of this study was performed to evaluate the plasma carnitine level in children with idiopathic epilepsy treated with old antiepileptic drugs [valproic acid and carbamazepine] and new antiepileptic drugs [lamotrigine and oxcarbazepine]. This study was carried out in Tanta University Hospital Pediatric Department, Neurology Unit. Fifty children with newly diagnosed idiopathic epilepsy were selected from those attending the pediatric neurology out-patient clinic. Thirty four [34] males and sixteen [16] females were enrolled in the study with the age range was 1-12 years with the mean age was [6.8 +/- 3 years]. Patients were grouped according to their antiepileptic treatment into: Group 1, twenty patients received valproic acid as monotherapy without any antiepileptic drugs treatment before. Group 2 ten patients received valproic acid as polytherapy after three months treatment with carbamazepine. Group 3, ten patients received lamotrigine as monotherapy, and group 4, ten patients received oxcarbazepine as monotherapy. Twenty healthy children served as control group with the age range was, 2-12 years with the mean age was [8 +/- 2 years]. Estimation of the plasma carnitine levels were done for all the studied groups. Group 1 and group 2 epileptic children, treated with valproic acid monotherapy and polytherapy had significantly lower plasma carnitine levels than the control group [P<0.05]. There was significant correlation between the age and the plasma carnitine in group 1 and group 2 epileptic children, the younger the age the more reduction in the plasma carnitine level Patients treated with valproic acid polytherapy had significantly lower plasma carnitine than patients treated with valproic acid monotherapy [P<0.05] .There was no significant difference between the plasma carnitine in children with epilepsy treated with oxcarbazepine and lamotrigine and the control group [P>0.05]. In conclusion, 1-Carnitine deficiency is not uncommon among children with epilepsy and is mainly linked to valproate therapy. 2-valproate may induce carnitine deficiency, but most cases are asymptomatic. 3-In contrast new-generation antiepileptic drugs probably do not cause carnitine deficiency. 4-These findings suggest a need to monitor serum carnitine levels in children treated with any of these drugs

Role of RANKL/OPG and L-carnitine systems in osteopenia/osteoporosis in children with B-thalassemia major

Bone disease in beta TM in the form of low bone mass remains a frequent, debilitating and poorly understood problem, even among well transfused and chelated patients. Receptor activator of nuclear factor kB ligand [RANKL] is a regulator of osteoclast formation and function, the activity of which appears to be a balance between interaction with its receptor and with antagonist binding protein osteoprotegerin [OPG]. L-carnitine enhanced osteoblasric activity as well. The objective is to define the role of RANKUOPG and L-carnitine systems-related bone loss in attempt for better management of osteopenialosteoporosis in beta TM children. The study included 69 I3TM children [45 males and 24 females, mean age 8.72 +/- 3. 70 ys and weight 22.84 +/- 7.04kg] attended the Children University Hospital [January-September 2008]. The patients were subdivided into 2 groups; one [n=34] received iron chelating [DFO] and the other group [n=35] did not receive any chelating drug. Fifteen healthy matched age, sex and BMI were included as controls. Serum OPG, sRANKL, L-carnitine, calcium, inorganic phosphorus and free fatty acids were measured by ELIZA and spectrophotometer. BMD, BMC and Z-score were measured by DEXA in 25 patients. There was a significantly lower serum level of OPG, L-carnitine and significantly higher sRANKL, sRANKL/OPG ratio, FFAs in patients than controls and in patients receiving DFO than those not receiving chelation. DEXA bone scanning detected osteopenia/osteoporosis in 12%and 88%of patients [mean Z score-3.38 +/- 1.19] with significantly higher osteoporosis in patients receiving DFO; more obvious in pelvis, L-spine, and femoral neck. L-carnitine correlated negatively with sRANKL, and positively with OPG. sRANKL correlated negatively with OPG. Z-score correlated negatively with age and positively with OPG. In conclusion, Reduced osteoblas tic activity and enhancing osteoclastic resorption are the basic mechanisms of bone loss in beta TM through decrease in L-carnitine and disturbed RANKL/OPG pathway. Recombinant OPG, L-carnitine and anti-RANKL supplement may be future agents that help in management of beta TM osteopenia/osteoporosis. Chelation with DFO seems to affect the bone density in such patients

Plasma-free carnitine levels in preterm infants with respiratory distress syndrome

Antenatal carnitine administration has been shown to induce fetal lung maturity by increasing pulmonary surfactant in animal and human studies. In this study, the aim was to investigate the status of carnitine in maternal and neonatal plasma of preterm infants with respiratory distress syndrome [RDS] in the first hours of life. We also aimed to characterize the carnitine status in these neonates with respect to sex, gestational age, birth weight, mode of delivery, and antenatal corticosteroid administration. Maternal plasma-free carnitine levels were determined before delivery and neonatal plasma-free carnitine levels were determined within 2 h of birth in preterm infants

possible cytoprotectiye effect of vitamins C and E and L-carnitine on monosodium glutamate induced oxidative stress in the rat

Monosodium glutamate [MSG] consumption has increased throughout the world as flavoring agent in cooking. However, consumption of a highly palatable diet has been shown to cause endothelial dysfunction, cardiovascular disease and various age-related degenerative disease processes long before development of any significant obesity. The major purpose of the present study was to investigate the effects of MSG on glucose metabolism, lipid profile and oxidative stress. A secondary purpose was to determine the possible protective effect of vitamin C, vitamin E and L-carnitine [LC], separately or in combination in MSG inducing oxidative stress in the rat. Sixty male albino rats weighing from 100-150 g were included in this study. Rats were divided into two main groups. Group 1, [10 rats] served as a control group for group II, and were received 1 ml of physiological saline [0.9%], daily subcutaneously [s.c.] for eight weeks. Group II: MSG -induced oxidative stress [50 rats] injected s.c. MSG in a dose of 200 mg/kg.b.wt. daily for eight weeks. Rats were further subdivided into A, B, C, D and E, each of ten rats. Group- A, received 1 ml of 2% gum acacia daily orally for eight weeks starting from the first day of MSG administration. Groups B, C, D treated with orally daily vitamin C in a dose of 100 mg/kg.b.wt. or vitamin E in a dose of 10 mg/kg.b.wt. or L-carnitine in a dose of 200 mg/kg.b.wt for the same previous duration. Group-E, received vitamins C, E and LC in the same previous doses and durations. S.C. injections of MSG produced significant increases in body weight, serum levels of glucose, insulin, leptin, total cholesterol, LDL-cholesterol and triglycerides, while HDL- cholesterol and Apo A 1 were significantly decreased. There were insignificant changes in serum iron, ferrritin and total iron binding capacity [TIBC] The levels of glutathione and the activities of both catalase and superoxide dismutase [SOD] in heart, liver and kidney were significantly decreased. In addition, there were significant decreases in both basal arid insulin stimulated glucose uptake by the soleus muscle and fatty tissue of the rat. Intake of vit. C or vit. E or LC, reduced the abnormalities in body weight, glucose metabolism, lipid profile and antioxidant enzymes. Also, they procducd a significant increase in both basal and insulin stimulated glucose uptake by the soleus muscle and fatty tissue of the rat. Still vit. C revealed better effects as compared to the corresponding groups. The combination of vit. C and E and LC was found to be the best as shown by the correction of glucose metabolism, lipid profile and highest levels of antioxidant enzymes as compared to the corresponding groups. MSG enhanced food intake. Overfeeding induced metabolic disorders associated with oxidative stress. The present study provides evidence that combination of antioxidant vitamins C and E with LC combat metabolic disorders and oxidative stress induced by MSG. Thus, sufficient dietary intake of these vitamins with LC is beneficial in combating MSG adverse effects

Increased serum and cardiac acyl-carnitine/free carnitine ratio during development of doxorubicin-induced cardiotoxicity

This study has been initiated to investigate whether cumulative doxorubicin [DOX] therapy alters serum and cardiac carnitine levels and if so, whether these alterations should be viewed as a mechanism and /or as an index during development of DOX-induced cardiotoxicity. To achieve the ultimate goal of this study, a total of 40 adult male Wistar albino rats were divided into 4 groups. In the first group, animals were injected intraperitoneally [I. P.] with normal saline [0.5 ml/200 gm body weight] and served as a normal control. Animals in the second to the fourth groups were injected every other day with DOX [3 mg/kg, I.P.], to obtain treatments with cumulative doses of 6 mg/kg [group 2], 12 mg/kg [group 3] and 18 mg/kg [group 4]. At 24 hours after receiving the last dose of DOX, animals were sacrificed serum as well as hearts were isolated and analyzed. DOX induced a significant and dose-dependent increase in serum creatine phosphokinase isoenzyme [CK-MB], lactate dehydrogenase [LDH], acyl-carnitine [AC]/free carnitine [FC] ratio and a significant decrease in serum free FC. In cardiac tissues, DOX induced a significant 46% and 63% decrease in FC after cumulative doses of 12 and 18 mg/kg, respectively. In contrast to FC, DOX induced a significant 70% and 81% increase in AC after cumulative doses of 12 and 18 mg/kg, respectively. Moreover, DOX treatment showed significant and dose-dependent decrease in adenosine triphosphate [ATP] level in cardiac tissues. In conclusion, data from this study suggest that: [1] Decreased myocardial carnitine level should be viewed as a mechanism during development of DOX cardiotoxicity, and [2] the parallel increase of serum AC/FC ratio and cardiotoxicity enzymatic indices, may point to the possible consideration of AC/FC ratio as a marker during development of DOX cardiotoxicity

Assessment of plasma carnitine and acylcarnitine levels in preeclampsia

Objective: carnitine plays a crucial role in fatty acids oxidation. The aim of the study is to assess plasma carnitine and acylcarnitines levels in preeclamptic womedas a measure of abnormal fatty acid oxidation

Methods: the study included 40 women with preeclampsia and 30 normotensive control women in the third trimester of pregnancy. Women with multiple pregnancy, chronic hypertension, diabetes mellitus and renal diseases were excluded. Plasma levels of free carnitine and acylcarnitines were measured with high performance liquid chromatography [HPLC]

results: total and free carnitines and acylcamitines were significantly increased in preeclamptic cases in comparison to the control group. A positive correlation was found between acylcarnitines and diastolic blood pressure [r=0.382, p= 0.018]

Conclusion: the significantly high plasma carnitine concentrations found in this study supports the hypothesis of abnormal fatty acid metabolism in the pathophysiology of preeclampsia. This may contribute to the endothelial cell dysfunction of preeclarnpsia

Plasma carnitine as a diagnostic marker at impaired left ventricular function in children with chronic rheumatic heart disease therapeutic role of L-carnitine

L-carnitine plays a major role in fatty acid oxidation and myocardial carnitine deficiency may cause malfunction of the head. Chronic rheumatic heart disease [RHD] can be associated with myocardial dysfunction. The objective of this study was to determine if plasma L-carnitine levels can serve as a diagnostic marker of impaired left ventricular [LV] function in children with chronic RHD, and to study the therapeutic role of L-carnitine in these patients. Forty cardiac patients with chronic RHD, with a mean age of 9.0 +/- 2.2 years, were enrolled in the study. They were classified into 2 groups: group I [with chronic RHD [20 patients], and group II with chronic RHD with congestive heart failure [CHF] [20 patients]. Twenty healthy children served as a control group. Plasma and urinary levels of L-carnitine were measured using Enzymatic UV test. Echocardiographic assessment of LV systolic function was performed [FS% and EF%]. Clinical and echocardiographic data were studied after 6 months of L-carnitine therapy in a group of 20 patients, compared to non-carnitine therapy group [20 patients]. The results showed that plasma and urinary L-carnitine levels were significantly higher in group II children with chronic RHD with CHF as compared to controls and group I patients [P<0.05]. There were significant negative correlations between plasma carnitine levels and echocardiographic indices of impaired LV systolic function [FS% and EF%] [P<0.05]. There was significant improvement of the clinical characteristics and echocardiographic LV systolic dysfunction, 6 months after L-carnitine therapy [P<0.05]. The elevated plasma [and urinary] L-carnitine levels in children with chronic RHD with CHF, can serve as a diagnostic marker of Impaired LV function. L-carnitine therapy may improve the clinical course and the LV systolic dysfunction in patients with chronic RHD

Serum carnitine levels in children with iron deficiency anemia

This study was designed to evaluate the carnitine serum levels as co-morbidity in apparently healthy children with iron deficiency anemia. Fifty four apparently healthy well nourished children [29 boys and 25 girls], their ages ranged from 9 months to 12 months with iron deficiency anemia were enrolled in the study. Twenty five healthy non anemic children with matched age and sex included as a control group. Malnourished children with iron deficiency anemia were excluded from the study. For all anemic children with suspected iron deficiency anemia as well as control group, we performed complete blood count [CBC] including measuring of hemoglobin [Hb] level, hematocrit% [Hct], mean corpuscular volume [MCV], mean corpuscular hemoglobin [MCH], mean corpuscular hemoglobin concentration [MCHC]; serum ferritin, serum iron [SI], and total iron binding capacity[TIBC]. Serum total carnitine levels were measured by spectrophotometric method. Hb, MCV, MCH, serum ferritin and serum iron were significantly lower in patients with iron deficiency anemia than in control non anemic group [p < 0.001]. Serum carnitine levels were significantly lower in children with iron deficiency anemia than in healthy control group [p<0.001]. There was a significant positive correlation between hemoglobin and serum carnitine [r=0.84; p<0.001]. Also, we found a significant positive correlation between serum iron, serum ferritin and serum carnitine [r=0.91; p<0.001 and r = 0.9; P<0.001, respectively]. In conclusion, iron is required for biosynthesis of carnitine. Low serum carnitine levels in these children may be due to iron deficiency. Therefore, iron fortification of the diets of children seems to be essential not only to prevent iron deficiency anemia but also to avoid other possible effects of iron deficiency in the growing children, such as secondary carnitine deficiency. However, additional studies are still needed to ascertain the frequency of carnitine deficiency and the need for carnitine supplementation in children with iron deficiency anemia

Serum carnitine and selenium levels in children with celiac disease.

BACKGROUND AND AIM: Celiac disease (CD) is a gluten-induced enteropathy that results in malabsorption of nutrients. We studied the serum levels of carnitine and selenium in children with CD. METHODS: Serum levels of free carnitine and selenium were studied in 30 children (mean age 8.1 [4.4] years) with CD and 30 age- and gender-matched healthy children. All patients had type 3 duodenal lesions. The mean (SD) serum levels of free carnitine and selenium were lower among patients with CD (24.5 [7.7] micromol/mL and 52.1 (12.9) micromol/mL, respectively) than among healthy controls (29.4 [9.2] and 65.1 [17.2] micromol/mL; p < 0.05 each). Levels were similar in children with and without diarrhea. CONCLUSIONS: Serum carnitine and selenium levels are decreased in children with CD, probably due to malabsorption.

Carnitina, niveles en el recién nacido y su relaión con toxremia gravídica / Carnitine concentration in the newborn and its relation with gravid toxemia

Introducción. La carnitina puede ejercer efectos relevantes para la medicina obstétrica y neonatal, por lo que garantizar un aporte adecuado de ésta durante el embarazo es importante para lograr un óptimo desarrollo del feto. En virtud de que la toxemia gravídica compromete en forma importante el flujo placentario, podría estar comprometido el paso de carnitina al feto, por lo que el presente estudio fue realizado con el propósito de comparar los niveles de carnitina plasmática obtenidos en recién nacidos de madres con toxemia gravídica (toxémicas) y los recién nacidos de madres sin toxemia gravídica (controles) y establecer la posible influencia de esta enfermedad sobre estos niveles. Material y métodos. Se determinaron los niveles plasmáticos de carnitina a 100 recién nacidos y a sus respectivas madres, 50 hijos de madres toxémicas y 50 controles. La determinación de los niveles plasmáticos de carnitina se realizó por el método enzimático descrito por Marquitz y Fitz. Resultados. Se observó una diferencia significativa en los niveles plasmáticos de carnitina entre los recién nacidos de ambos grupos, siendo estos niveles menores en el grupo de toxémicas (16.16 nM/mL). No se evidenció diferencia significativa en la carnitinemia de las madres. Existe correlación entre los niveles de carnitina maternos y del recién nacido en ambos grupos, pero con coeficientes de correlación distintos. Conclusiones. La toxemia gravídica se asocia a una disminución de los niveles plasmáticos de carnitina en el recién nacido, aunque se mantiene la correlación con los niveles maternos