Novel OCTN2 mutations: No genotype–phenotype correlations: Early carnitine therapy prevents cardiomyopathy

Primary systemic carnitine deficiency or carnitine uptake defect (OMIM 212140) is a potentially lethal, autosomal recessive disorder characterized by progressive infantile‐onset cardiomyopathy, weakness, and recurrent hypoglycemic hypoketotic encephalopathy, which is highly responsive to L ‐carnitine therapy. Molecular analysis of the SLC22A5 (OCTN2) gene, encoding the high‐affinity carnitine transporter, was done in 11 affected individuals by direct nucleotide sequencing of polymerase chain reaction products from all 10 exons. Carnitine uptake (at Km of 5 μM) in cultured skin fibroblasts ranged from 1% to 20% of normal controls. Eleven mutations (delF23, N32S, and one 11‐bp duplication in exon 1; R169W in exon 3; a donor splice mutation [IVS3+1 G > A] in intron 3; frameshift mutations in exons 5 and 6; Y401X in exon 7; T440M, T468R and S470F in exon 8) are described. There was no correlation between residual uptake and severity of clinical presentation, suggesting that the wide phenotypic variability is likely related to exogenous stressors exacerbating carnitine deficiency. Most importantly, strict compliance with carnitine from birth appears to prevent the phenotype. © 2002 Wiley‐Liss, Inc.     

Correlation between genotype,metabolic data,and clinical presentation in carnitine palmitoyltransferase 2 (CPT2) deficiency

Carnitine palmitoyltransferase 2 (CPT2) deficiency, the most common inherited disease of the mitochondrial long-chain fatty acid (LCFA) oxidation, may result in distinct clinical phenotypes, namely a mild adult muscular form and a severe hepatocardiomuscular disease with an onset in the neonatal period or in infancy. In order to understand the mechanisms underlying the difference in severity between these phenotypes, we analyzed a cohort of 20 CPT2-deficient patients being affected either with the infantile (seven patients) or the adult onset form of the disease (13 patients). Using a combination of direct sequencing and denaturing gradient gel electrophoresis, 13 CPT2 mutations were identified, including five novel ones, namely: 371G>A (R124Q), 437A>C (N146T), 481C>T (R161W), 983A>G (D328G), and 1823G>C (D608H). After updating the spectrum of CPT2 mutations (n=39) and genotypes (n=38) as well as their consequences on CPT2 activity and LCFA oxidation, it appears that both the type and location of CPT2 mutations and one or several additional genetic factors to be identified would modulate the LCFA flux and therefore the severity of the disease.     

Brief intense exercise followed by passive recovery modifies the pattern of fuel use in humans during subsequent sustained intermittent exercise

The role of work period duration as the principal factor influencing carbohydrate metabolism during intermittent exercise has been investigated. Fuel oxidation rates and muscle glycogen and free carnitine content were compared between two protocols of sustained intermittent intense exercise with identical treadmill speed and total work duration. In the first experiment subjects (n=6) completed 40 min of intermittent treadmill running involving a work : recovery cycle of 6 : 9 s or 24 : 36 s on separate days. With 24 : 36 s exercise a higher rate of carbohydrate oxidation approached significance (P=0.057), whilst fat oxidation rate was lower (P ≤ 0.01) and plasma lactate concentration higher (P ≤ 0.01). Muscle glycogen was lower post‐exercise with 24 : 36 s (P ≤ 0.05). Muscle free carnitine decreased (P ≤ 0.05), but there was no difference between protocols. In the second experiment a separate group of subjects (n=5) repeated the intermittent exercise protocols with the addition of a 10‐min bout of intense exercise, followed by 43 ± 5 min passive recovery, prior to sustained (40 min) intermittent exercise. For this experiment the difference in fuel use observed previously between 6 : 9 s and 24 : 36 s was abolished. Carbohydrate and fat oxidation, plasma lactate and muscle glycogen levels were similar in 6 : 9 s and 24 : 36 s. When compared with the first experiment, this result was because of reduced carbohydrate oxidation in 24 : 36 s (P ≤ 0.05). There was no difference, and no change, in muscle free carnitine between protocols. A 10‐min bout of intense exercise, followed by 43 ± 5 min of passive recovery, substantially modifies fuel use during subsequent intermittent intense exercise.     

Significance of Levocarnitine Treatment in Dialysis Patients

Carnitine is a naturally occurring amino acid derivative that is involved in the transport of long-chain fatty acids to the mitochondrial matrix. There, these substrates undergo β-oxidation, producing energy. The major sources of carnitine are dietary intake, although carnitine is also endogenously synthesized in the liver and kidney. However, in patients on dialysis, serum carnitine levels progressively fall due to restricted dietary intake and deprivation of endogenous synthesis in the kidney. Furthermore, serum-free carnitine is removed by hemodialysis treatment because the molecular weight of carnitine is small (161 Da) and its protein binding rates are very low. Therefore, the dialysis procedure is a major cause of carnitine deficiency in patients undergoing hemodialysis. This deficiency may contribute to several clinical disorders in such patients. Symptoms of dialysis-related carnitine deficiency include erythropoiesis-stimulating agent-resistant anemia, myopathy, muscle weakness, and intradialytic muscle cramps and hypotension. However, levocarnitine administration might replenish the free carnitine and help to increase carnitine levels in muscle. This article reviews the previous research into levocarnitine therapy in patients on maintenance dialysis for the treatment of renal anemia, cardiac dysfunction, dyslipidemia, and muscle and dialytic symptoms, and it examines the efficacy of the therapeutic approach and related issues.     

Long-chain 3-hydroxyacyl-coenzyme A dehydrogenase deficiency—diagnosis,plasma carnitine fractions and management in a further patient

Long-chain 3-hydroxyacyl-coenzyme A dehydrogenase (LCHAD), the third enzyme of the mitochondrial -oxidation pathway, carries out the dehydrogenation of 3-hydroxyacyl-CoA compounds of 12–18 carbon length. To date only nine cases of LCHAD deficiency have been documented. We report a further patient who as a neonate developed non-specific gastro-intestinal symptoms and at 5 months of age cardiomyopathy, recurrent hypoketotic hypoglycaemia and gross alterations of plasma carnitine fractions. Dietary management with medium chain triglycerides led rapidly to clinical improvement. There was a close correlation between the clinical condition, plasma carnitine fractions and cardiac function. At 2 years of age she is developing normally.     

卡尼汀的生物学作用及在血液透析患者中的临床应用

卡尼汀是长链脂肪酸进入线粒体进行氧化所必需的一种物质，其左旋异构体具有生物活性。正常人可通过饮食摄入和肝、肾合成产生卡尼汀，但尿毒症血液透析患者由于肾脏内源性合成卡尼汀减少，血液透析过程中大量清除，大多存在时间依赖性的卡尼汀缺乏。通过静脉注射、口服、透析液补充外源性左旋卡尼汀有利于纠正脂代谢紊乱，减少促红细胞生成素用量，改善骨骼肌症状和营养状态，提高生活质量。     

Carnitine depletion during total parenteral nutrition despite oral L-carnitine supplementation

Carnitine (CAR) plays an important role in the β-oxidation of fatty acids. Less attention, however, has been paid to CAR compared to other nutrients even in total parenteral nutrition (TPN). To examine CAR metabolism during TPN and the effect of simultaneous oral L-CAR supplementation on CAR levels, the blood CAR level was measured in a 3-year-old boy receiving long-term TPN because of short bowel syndrome. Both the total and acyl CAR in the serum were evaluated under various nutritional conditions including oral supplementation of L-CAR. Low CAR concentrations were observed especially when lipid containing TPN regimens were in place. Oral L-CAR supplementation was not sufficient to restore the low CAR levels in the present index patient even when the dose was increased to 120 mg/kg in accordance with the result of the L-CAR absorption test that revealed poor intestinal absorption of this nutrient. Moreover, a markedly low CAR level was measured during the onset of sepsis in the patient, and the blood CAR was depleted when lipid metabolism was activated by lipid loading or sepsis. To date, the late effects of CAR depletion on child growth have not been well examined. It is recommended that the blood CAR level be maintained at normal levels before any prominent manifestations of the deficiency have developed. The intravenous administration of CAR appears to be necessary to supply a sufficient amount of CAR for patients with severe malabsorption.     

血液透析对游离肉碱的清除及影响因素的研究

目的　研究维持性血液透析 (MHD)患者血浆游离肉碱 (FC)水平以及透析对FC清除的影响。方法　应用酶法检测MHD组血浆和透析液中FC水平及对照组尿中FC水平。结果　MHD患者透前血浆FC水平明显低于正常人 (P <0 .0 1)。透析后 ,血浆FC水平较透析前下降 70 %。每周MHD患者透析所致的FC丢失量远远超过对照组尿液排出量 (P <0 .0 1)。透析过程中 ,FC在第 1小时清除最多。透析中FC的清除量与血浆FC浓度呈正相关 (R =0 .5 1,P =0 .0 1)。应用不同透析器及其复用对FC的清除无显著差异。结论　透析中丢失是MHD患者肉碱缺乏的重要原因。应用不同透析器及其复用对FC清除无明显影响。     

肉碱和长链脂肪醇对小鼠大脑乙酰胆碱的影响

目的　探讨肉碱和长链脂肪醇对小鼠大脑乙酰胆碱 (Ach)含量的影响。方法　用析因实验设计方法将BALB/C成年雄性小鼠随机分为标准对照、高脂对照、肉碱、长链脂肪醇、肉碱和长链脂肪醇合用共 5组 ,标准对照组饲标准饲料 ,其它各组饲含 2 0 %油脂的饲料 ,肉碱 [剂量 50mg/ (kg·鼠重 ) ]和长链脂肪醇 [剂量 50mg/ (kg·鼠重 ) ]以0 0 9%的剂量加到饲料中连续喂养 3 0d后 ,称体重、脑重 ,计算脑体比 ,用碱性羟胺比色法测大脑组织中Ach含量。结果　肉碱和长链脂肪醇对成鼠体重、脑重、脑体比无显著影响 (P >0 0 5) ;高脂饲料组小鼠脑内Ach含量较对照组明显降低 (P <0 0 5) ,长链脂肪醇可引起高脂动物脑内Ach含量升高 (P <0 0 5) ,但与正常饲料组无明显差异 (P >0 0 5) ;肉碱引起脑内Ach含量明显增多 (P <0 0 5) ;但两者联合作用时有交互作用 (P <0 0 1) ,且脑内Ach含量约等于者单独作用的差值。结论　肉碱、长链脂肪醇能分别促进小鼠大脑Ach含量的增加 ,但联合作用时表现为拮抗