Carnitine analysis in pterygium / Análise de carnitina no pterígio

ABSTRACT Purpose: The aim of the present study was to measure the free carnitine and acylcarnitine levels in pterygium tissue and normal conjunctival tissue at the metabolomics level using tandem mass spectrometry. Methods: In this prospective, clinical randomized study, pterygium tissues and normal conjunctival tissues taken during pterygium excision with autograft were compared regarding their free carnitine and acylcarnitine profiles. After tissue homogenization, carnitine levels were measured using tandem mass spectrometry. The data were statistically analyzed with the Wilcoxon signed-rank test. Results: Pterygium and normal conjunctival tissue samples from a single eye of 29 patients (16 females, 13 males; mean age, 54.75 ± 11.25 years [range, 21-78 years]) were evaluated. While the free carnitine (C0) level was significantly high in the pterygium tissue (p<0.001), acylcarnitine levels were significantly high in some esterized derivatives (C2, C5, C5:1, C5DC, C16:1, C18, methylglutarylcarnitine) (p<0.05). No statistically significant difference was determined for the other esterized derivatives (p>0.05). Conclusion: That the carnitine levels in pterygium tissue were higher suggests that acceleration of cell metabolism developed secondary to chronic inflammation and the premalignant characteristics of pterygium tissue. High carnitine levels may also effectively suppress the apoptosis process. The data reported in our study indicate that further, more extensive studies of the carnitine profile could help clarify the pathogenesis of pterygium.

RESUMO Objetivo: O objetivo deste estudo foi medir os níveis de carnitina livre e acil-carnitina a nível metabolómico com espectrometria de massa em tandem no tecido do pterígio e no tecido conjuntivo normal. Método: Neste estudo prospetivo, clínico e aleatório, os tecidos de pterígio e os tecidos normais de conjuntiva, retirados durante a cirurgia de pterígio com autoenxerto, foram comparados em relação ao perfil de carnitina livre e de acil-carnitina. Após a homogeneização dos tecidos, os níveis de carnitina foram medidos por espectrometria de massa em tandem. A análise estatística dos dados foi realizada com o teste dos postos sinalizados de Wilcoxon. Resultados: A avaliação foi feita através de amostras de tecido pterígio e de conjuntiva normal de um único olho de 29 pacientes (16 mulheres, 13 homens). A média de idade dos pacientes foi de 54,75 ± 11,25 anos (faixa dos 21 aos 78 anos). Enquanto o nível de carnitina livre (C0) foi significativamente elevado no tecido pterígio (p<0,001), os níveis de acil-carnitina foram significativamente elevados em alguns derivados esterificados (C2, C5, C5: 1, C5DC, C16:1, C18, metilglutaril carnitina) (p<0,05). Não foi determinada uma diferença estatisticamen te significante noutros derivados esterificados (p>0,05). Conclusão: Os níveis mais elevados de carnitina no tecido do pterígio sugerem que a aceleração do metabolismo celular se tenha tornado secundária com o efeito da inflamação crónica e o caráter pré-maligno do tecido do pterígio. Os níveis elevados de carnitina também podem ser eficazes na supressão do processo de apoptose. Os dados obtidos no estudo indicam que estudos mais extensivos do perfil da carnitina contribuiriam para o esclarecimento da patogénese do pterígio.

Three cases of carnitine-acylcarnitine transposase deficiency and literature review / 中国新生儿科杂志

Objective To study the clinical features,biochemical characteristics and gene mutations of patients with carnitine-acylcarnitine translocase deficiency (CACTD).Method The clinical data,biochemical markers and gene mutations of three cases with CACTD admitted our hospital in 2017 were retrospectively analyzed.The related literatures were searched from China national knowledge infrastructure,wanfang database,PubMed,national center for biotechnology information and Embase using keywords "neonate","infant","carnitine-acylcarnitine deficiency","carnitine-acylcarnitine translocase",and SLC25A20"(up to April 2018).Result (1) Three cases (2 boys and 1 girl) with CACTD were full-term infants without asphyxia after birth.The mothers had no abnormal pregnancy,and the parents had no consanguinity.All the patients had poor response and severely hypoglycemia 15～20 hours after birth.Hyperammonemia,elevated liver enzymes and creatine kinase,severe dicarboxylic aciduria,significantly increased level of long-chain acylcarnitine,and significantly decreased concentration of free carnitine were observed in all 3 patients.Significantly decreased serum ketone body was observed in 2 cases.All of them had recurrent atrioventricular block and ventricular tachycardia requiring repeated electrocardioversion,lidocaine,and amiodarone treatment.Arginine,carnitine and special formula with low fat and high medium-chain-triglyceride were given to two infants.Two infants died of cardiorespiratory failure at 3-day and 8-day of life,respectively.The other infant's clinical condition improved significantly.However,he was discharged from our NICU at the request of his parents.Gene analysis revealed that compound heterozygous mutations c.199-10T＞G and IVS7-9_16 ins (a possible novel mutation) were detected in the SLC25A20 gene of case 2.Homozygous mutation c.199-10T＞G was identified in the SLC25A20 gene of case 3 whose parents both carried this mutation.(2) A total of 17 articles and 50 cases were retrieved and analyzed.A total of 40 mutations were found in the SLC25A20 gene.Homozygous mutations were found in 23 cases,and compound heterozygous mutations were found in 27 cases.The mutation of c.199-10T＞G was the most common mutation and occurred 22 times in the patients from Asia population.Other mutations were found less than 6 times.The review showed that the most common clinical features included hypoketotic hypoglycemia,hyperammonemia,elevated liver enzymes and creatine kinase,remarkable dicarboxylic aciduria,significantly increased level of long-chain acylcarnitine,significantly decreased free carnitine,arrhythmia and cardiomyopathy.Mostly,the onset of symptoms was within 1 week after birth (88％,44/50).The mortality was 69.8％ (30/43).Most patients died within the first year of their life.Conclusion Early recognition,early diagnosis and prompt treatment are crucial for CACTD patients.Gene analysis is a reliable diagnostic method.The mutation of c.199-10T＞G is the most common SLC25A20 mutation reported in Asia population.Hypoketotic hypoglycemia is an early sign of this disease.Families with a proband need prenatal diagnosis during the second pregnancy.

Carnitine Deficiency in Chinese Children with Epilepsy on Valproate Monotherapy

Objective: To explore the incidence and independent risk-factors of secondary carnitinedeficiency in Chinese children with epilepsy on valproate monotherapy. Methods: The freecarnitine and acylcarnitines levels in 299 children with epilepsy on valproate monotherapybetween June 2014 and September 2015 were compared with age- and sex-matched 299healthy controls. Results: Children with valproate monotherapy had lower free carnitinelevels [23.86 (10.60) µmol/L] than the controls [36.37 (9.37) µmol/L] (P<0.01). Mostacylcarnitines were significantly lower in children with valproate monotherapy than controls.63 children (21.1%) with epilepsy had carnitine deficiency; 54 were asymptomatic. Femalegender (OR 2.1), high alanine aminotransferase levels (OR 1.0) and long duration of VPAtreatment (1-12 mo) (OR 1.9) were independent risk factors for secondary carnitinedeficiency induced by VPA. Conclusions: Carnitine deficiency with valproate is more likely infemales, those with transaminitis, and those receiving the drug for 1-12 months

Value of blood mass spectrometry analysis in differentiating decompensated liver cirrhosis patients / 中国医师进修杂志

Objectives To compare the difference of blood mass spectrometry analysis between decompensated liver cirrhosis patients and compensated liver cirrhosis patients. Methods Thirty-four patients with decompensated liver cirrhosis and 47 patients with compensated liver cirrhosis were selected. All patients were analyzed by blood mass spectrometry, and the detected 93 parameters included amino acids, acylcarnitines and some of their ratios. Results After multivariate analysis using the 93 parameters, the two groups could be differentiated clearly. There were 32 parameters that contributed to the separation, which included 4 ratios. Logistic analysis showed that alanine, tetradecanoyl diacylcarnitine, tetradecenoylcarnitine, proline and serine could be used to separate the two groups sufficiently. Receiver operating characteristic curve analysis indicated that the area under the curve generated using the 5 parameters could reach to 0.99. Except alanine, the other 4 metabolites were all increased in the decompensated patients. Conclusions Blood mass spectrometry analysis could be used to differentiate decompensated and non- decompensated cirrhosis patients. The significantly changed metabolites might provide valuable hint in pathological study of the disease.

The SLC22A5 genetic analysis in Chinese patients with systemic primary carnitine deficiency / 中华内分泌代谢杂志

Objective To investigate the clinical and biochemical metabolic features of 12 patients with systemic primary carnitine deficiency(CDSP) and to identify the SLC22A5 gene mutation types of the disease. Method The clinical and biochemical data were collected by retrospective analysis. DNA direct sequencing and multiplex ligation dependent probe amplification(MLPA)were applied for SLC22A5 gene analysis. Result Among 12 patients with CDSP, 3 cases had evident infection factors, 6 cases with convulsions, 5 cases manifested liver hypertrophy, 8 cases with hyperammonemia, and 9 cases showed myocardial damage. All CDSP patients were detected biallelic pathogenic mutation in SLC22A5 gene by direct sequencing. The gene types include IVS2+1G>T, c.3G>T(p.Met1Ile), c.760C>T(p.Arg254X), c.1400C>G(p.Ser467Cys), c.844dupc(p.Arg282fs), c.338G>A(p.Cys113Tyr), c.51C>G(p.Phe17Leu), c.659A>T(p.Glu220Val), and c.1365dupC(p.Thr456fs). c.659A>T(p.Glu220Val) and c.1365dupC(p.Thr456fs)are novel mutations. One female patient was maternal CDSP, her child had abnormal newborn screening. The allele frequency of c.760C>T(p.Arg254X) and c.1400C>G(p.Ser467Cys) were 37.5%(9/24)and 29.2%(7/24)respectively. The MLPA test results of all patients were negative. Conclusion The clinical manifestations are complex and various in patients with CDSP. Point and small InDel(insertions/deletions)mutation constitute the major alteration in SLC22A5 gene. c.1400C>G(p.Ser467Cys) might be another prevalence mutation type in Chinese CDSP patient.

Metabonomic study of newborn infants with intrauterine growth restriction / 中华实用儿科临床杂志

Objective To compare the differences in metabolites between newborns with intrauterine growth restriction (IUGR) and appropriate for gestational age (AGA) in order to understand the changes in metabolites of newborns with IUGR and explore the possible metabolic mechanism of tissue and organ damages in patients with IUGR,with the ultimate goal of providing the basis for clinical intervention.Methods A total of 45 newborns with IUGR and 56 AGA newborns who were hospitalized in the Neonatal Intensive Care Unit of Bayi Children's Hospital,the General Hospital of the Chinese People's Liberation Army between July 2009 and June 2015 and who underwent metabolic disease screening were enrolled in this study.The differences in of 21 amino acids and 55 carnitines in peripheral blood,as well as the changes in the ratios of free carnitine and acylcarnitine to total carnitine,were compared.Results (1)According to the comparison of birth weights (＜ 3rd percentile,3rd-＜ 5th percentile,5th-＜ 10th percentile,and 10th-90th percentile),peripheral blood of the IUGR newborns with birth weight ＜ 3rd percentile contained lower concentrations of alanine (F =2.94,P =0.03),homocysteine (F =3.83,P =0.01),methionine (F =2.88,P =0.04),ornithine(F =3.32,P =0.02),serine (F =3.09,P =0.03) and tyrosine (F =4.76,P =0.00) than those of the AGA newborns.In the peripheral blood of the IUGR newborns with birth weight of 3rd-＜ 5th percentile,the diversity of alanine concentrations showed compensatory increase,and their alanine concentrations were higher than those of the AGA newborns.(2) Metabolites also had significant differences in different gestational age groups:the concentrations of alanine (t =2.423,P =0.026),proline (t =2.470,P =0.023),and 14-carbon acylcarnitine (t =-2.870,P =0.010) in premature was higher than those in full-term newborns,but the concentration of 26-carbon acylcarnitine (t =-2.189,P =0.041) was lower than full-term ones;the concentrations of alanine (t =2.354,P =0.022),glutamine (t =2.520,P =0.015),pipecolic acid (t =2.017,P =0.049),proline (t =2.204,P =0.032) in premature AGA newborns were higher than those in full-term ones,but the concentrations of homocysteine (t =-2.624,P =0.011),seven carbon acylcarnitine(t =-2.403,P =0.020),and ten carbon acylcarnitine (t =-5.739,P =0.000) were lower than those of full-term AGA newborns;the concentrations of homocysteine (t =-2.421,P =0.020),decanogl carnitine(t =-2.181,P =0.035),methyl propylene acyl carnitine (t =-2.373,P =0.022),pentyl acyl carnitine (t =-2.165,P =0.036),decyl acyl carnitine (t =-4.148,P =0.000),hydroxyl acetyl carnitine (t =-2.097,P =0.042),hydroxyl cetyl acylcarnitine (t =-2.446,P =0.019) in premature IUGR were higher than those in fullterm IUGR newborns;but the concentrations of arginine (t =2.167,P =0.036),glutamic acid (t =2.469,P =0.018),histidine (t =2.718,P =0.009),leucine/isoleucine (t =3.938,P =0.000),ornithine (t =4.264,P =0.000),serine (t =2.647,P =0.011),threonine (t =2.311,P =0.026),tryptophan (t =4.040,P =0.000),valine (t =2.700,P =0.01),7-carbon acylcarnitine (t =-2.44 1,P =0.019),18-carbon diene carnitine (t =2.449,P =0.018),capric acylcarnitine(t =-4.148,P =0.000) and hydroxyl acetyl carnitine (t =-2.097,P =0.042) were lower than those in full-term IUGR newborns.(3) For AGA newborns,metabolites had no differences between male and female (P ＞ 0.05);however,for newborns with IUGR,metabolites significantly differed between male and female,and the concentrations of aspartic acid(t=2.521,P =0.016),glutamate(t =-2.175,P =0.035) in male IUGR were lower than those in female newborns with IUGR,but the concentration of 26-carbon carnitine (t =2.231,P =0.031) was higher than that in female group.(4) Birth weight had no significant effect on free carnitine concentration or on the ratios of free carnitine and acylcarnitine to total carnitine(all P ＞ 0.05).Conclusions IUGR infants exhibit significant abnormalities in amino acid and acylcarnitine metabolism,especially those with birth weight ＜ 3rd percentile.With the increase of birth weight,amino acids and acylcarnitines showed compensatory increases or decrease,and when birth weight reached the 10th percentile,the newborns with IUGR were close to the AGA newborns.

Acylcarnitine profile in children of malnutrition / 中华临床营养杂志

Objective To summarize the acylcarnitine profile in children with malnutrition,with an attempt to distinguish it from those of medium-chain acyl-CoA dehydrogenase (MCAD) deficiency,multiple acylCoA dehydrogenase (MAD) deficiency,or glutaric aciduria type Ⅱ (GA Ⅱ).Methods Thirteen pediatric patients with malnutrition and 214 children of the same age but without malnutrition,which was set as the control group,were included in this study.The blood samples were collected at admission,and the concentration of carnitine and acylcarnitines were measured in bloodspots by tandem mass spectrometry using samples nnderivatized.Results The concentrations of acylcarnitines which were involved in fatty acid oxidation,including octadecanoyl (C18) to acetyl (C2) acylcarnitines and ketonic acylcarnitines,were higher in malnutrition group than in the control group.Particularly,the concentration of decanoyl acylcarnitine (C10) in the malnutrition group was (0.203 ±0.105) μmol/L,which was out of the normal rang (0-0.200 μmol/L),was significantly higher than that [(0.054 ±0.030) μmol/L] in the control group (P ＜0.001).There was no significant difference in the concentrations of acylcarnitines [e.g.propionyl (C3),isovaleryl (C5),3-hydroxy-isovaleryl (C5OH),and glutaryl (C5DC) acylcamitines] involved in amino acid decomposition between the malnutrition and control groups.Conclusions The concentrations of acylcarnitines related to fatty acid oxidation elevate in children with malnutrition.In particular,the medium-chain acylcarnitines C10 is out of the normal range,which can be used to differentiate malnutrition from MCAD and MAD.

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.

Hiperinsulinismo na infância: quando apenas uma dosagem de insulina não é suficiente / Hyperinsulinism in infancy and childhood: when an insulin level is not always enough

INTRODUÇÃO: A hipoglicemia em bebês e crianças pode causar convulsões, atraso de desenvolvimento e dano cerebral permanente. O hiperinsulinismo (HI) é a causa mais comum de hipoglicemia, seja transitória ou permanente. A HI é caracterizada pela secreção inadequada de insulina, o que resulta em hipoglicemia persistente, de leve a grave. As diferentes formas de HI representam um grupo de doenças clínica, genética e morfologicamente heterogêneo. CONTEÚDO: Hiperinsulinismo congênito está associado às mutações de SUR-1 e Kir6.2, glucoquinase, glutamato desidrogenase, 3-hidroxiacil-CoA desidrogenase de cadeia curta e expressão ectópica de SLC16A1 na membrana plasmática das células beta. O HI pode estar associado ao estresse perinatal, como asfixia do nascimento, toxemia materna, prematuridade ou retardo do crescimento intra-uterino, resultando em hipoglicemia neonatal prolongada. Mimetismo de HI neonatal inclui pan-hipopituitarismo, hipoglicemia induzida por fármaco, insulinoma, anticorpos antiinsulina e estimuladores do receptor de insulina, síndrome de Beckwith-Wiedemann e distúrbios congênitos de glicosilação. Exames laboratoriais para HI podem incluir quantificação de glicose, insulina, β-hidroxibutirato, ácidos graxos, amônia e perfil de acilcarnitinas plasmáticos, além de ácidos orgânicos urinários. Os exames genéticos estão disponíveis em laboratórios comerciais para os genes sabidamente associados à hiperinsulinemia. Testes de resposta insulínica aguda (RIA) são úteis na caracterização fenotípica. Exames de imagem e histológicos também estão disponíveis para diagnosticar e classificar o HI. O objetivo do tratamento de crianças com HI é prevenir os danos cerebrais da hipoglicemia, mantendo níveis de glicose plasmática acima de 70mg/dl por terapia farmacológica ou cirúrgica. CONCLUSÃO:A terapêutica do HI requer abordagem multidisciplinar que inclui endocrinologistas pediátricos, radiologistas, cirurgiões e patologistas, os quais são treinados para diagnosticar..

BACKGROUND: Hypoglycemia in infants and children can lead to seizures, developmental delay, and permanent brain damage. Hyperinsulinism (HI) is the most common cause of both transient and permanent disorders of hypoglycemia. HI is characterized by dysregulated insulin secretion, which results in persistent mild to severe hypoglycemia. The various forms of HI represent a group of clinically, genetically, and morphologically heterogeneous disorders. CONTENT: Congenital hyperinsulinism is associated with mutations of SUR-1 and Kir6.2, glucokinase, glutamate dehydrogenase, short-chain 3-hydroxyacyl-CoA dehydrogenase, and ectopic expression of SLC16A1 on β-cell plasma membrane. Hyperinsulinism may be associated with perinatal stress such as birth asphyxia, maternal toxemia, prematurity or intrauterine growth retardation, resulting in prolonged neonatal hypoglycemia. Mimickers of hyperinsulinism include neonatal panhypopituitarism, drug-induced hypoglycemia, insulinoma, antiinsulin and insulin-receptor stimulating antibodies, Beckwith-Wiedemann Syndrome, and congenital glycosylation disorders. Laboratory testing for hyperinsulinism may include quantification of blood glucose, plasma insulin, plasma β-hydroxybutyrate, plasma fatty acids, plasma ammonia, plasma acylcarnitine profile and urine organic acids. Genetic testing is available at commercial laboratories for genes known to be associated with hyperinsulinism. Acute insulin response (AIR) tests are useful in phenotypic characterization. Imaging and histological tools are also available to diagnose and classify hyperinsulinism. The goal of treatment in infants with hyperinsulinism is to prevent brain damage from hypoglycemia by maintaining plasma glucose levels above 700 mg/l (70 mg/dl) through pharmacologic or surgical therapy. SUMMARY: The treatment of hyperinsulinism requires a multidisciplinary approach that includes pediatric endocrinologists, radiologists, surgeons, and pathologists who trained to diagnose...

Fatty acid oxidation disorders / 한양의대학술지

Inborn errors of fatty acid mitochondrial oxidation (FAOD) have drawn considerable attention in recent years because of rapid pace of discovery of new defects and an ever-increasing spectrum of clinical phenotypes. This review describes a clinical and biochemical phenotypes, diagnosis and treatment of FAOD. Some of FAOD can not be detected by conventional biochemical investigations, even when a patient is symptomatic with fasting intolerance or functional failure of fatty acid dependent tissue (s). Diagnosis must ultimately be based on direct assay of the involved enzyme, however, preliminary indicators may come from determination of carnitine and intermediate metabolites in plasma, profiling of urine organic acid, and radioisotopic screening assays with lymphocytes or cultured fibroblasts. We are faced with the following major challenges: whether to include FAOD in newborn screening programs, the investigation of the rules played by individual disorders in maternal complication during pregnancy, sudden and unexpected death in early life, and pediatric acute/fulminant liver failure.