Clinical and molecular studies on in born errors of ketone body metabolism

伴有轻度突变的SCOT缺陷患者：我们从临床和分子水平分析了3个日本家系的SCOT缺陷患者。我们发现，具有“轻微”突变的SCOT缺乏症患者没有表现出病理性的永久性酮病。这表明，如果患者没有永久性酮症，我们不能排除SCOT缺乏症(提交)。T2缺乏症患者启动子甲硫氨酸密码子突变的特征：我们在GK30中发现了启动密码子突变c.2T&gt；C(另一突变为149delC)。我们构建了9种可能的启动子密码子错义突变的表达载体，并分析了在突变启动子密码子上翻译起始的效率。虽然突变的效率不同，但所有突变的甲硫氨酸密码子都作为启动子密码子发挥作用。因此，我们得出结论：启动子密码子突变导致了T2缺乏症的轻微突变(2003年)。T2缺乏症…的特征更多具有“轻微”突变的患者：我们分析了5名日本T2缺陷患者的表型/基因相关性，揭示了具有“轻微”突变的T2缺陷患者的特征。我们发现，在“轻度”和“严重”突变患者之间，酮症酸中毒危象的发生频率和严重程度没有显著差异。然而，在非发作期条件下，在“轻微”突变的患者中，尿液有机酸和血酰卡尼汀谱的异常是微乎其微的，如果在非发作期条件下进行这些分析，它们可能被忽略为正常(2003)。我们还提出了一种可能性，即通过与tyglyl-CoA(提交的)Scot，3HBD基因敲除小鼠的偶联分析，有“轻微”突变的患者被误诊为正常：我们克隆了构建基因敲除构建体所需的基因组片段，并仍在构建中。2-甲基-3-羟基丁酸脱氢酶缺乏症的酶检测方法的建立及其临床分析：我们利用成纤维细胞和淋巴细胞建立了该酶的酶检测方法。我们没有在之前排除T2缺乏症的细胞系中发现这种酶缺陷。较少

SCOT deficient patients with "mild" mutation : We analyzed 3 Japanese families with SCOT deficient patients at the clinical and molecular level. We disclosed that SCOT deficient patients with "mild" mutations do not show a pathognomonic permanent ketosis. This indicates that we can not exclude SCOT deficiency if patients do not have permanent ketosis (Submitted).Characterization of mutations at the initiator methionine codon in T2 deficiency : We identified an initiator codon mutation, c.2T>C, heterozygously in GK30 (The other mutation was 149delC). We made expression vectors for 9 possible missense mutations at the initiator codon and analyzed the efficiency of translation initiation at the mutant initiator codons. Although the efficiencies were variable among the mutations, all the mutant initiator methionine codons were functioned as the initiator codon. Hence we concluded that initiator codon mutations result in "mild" mutations in T2 deficiency (2003).Characterization of T2 defici … More ent patients with "mild" mutations : We analyzedphenotype/genotype correlation in 5 Japanese T2 deficient patients and revealed characters for T2 deficient patients with "mild" mutations. We found no significant differences in the frequency and severity of ketoacidotic crises between patients "mild" and "severe" mutations. However, during non-episodic condition, abnormalities in urinary organic acid and blood acylcarnitine profiles was subtle in patients with "mild" mutations and they, can be missed as normal if these analyses are performed during non-episodic condition (2003). We also raised a possibility that patients with "mild" mutations were mis-diagnosed as normal by a coupled assay with tyglyl-CoA (submitted)SCOT, 3HBD knockout mice : We cloned genomic fragments necessary for construction of knockout constracts and are still making constructs.Establishment of enzyme assay for 2-methyl-3-hydroxybutyrate dehydrogenase deficiency and its clinical analysis : We established enzyme assay method for this enzyme using fibroblasts and lymphocytes. We did not find this enzyme defect in cell lines which were previously ruled out T2 deficiency. Less