PURPOSE: Seizure associated with fever may indicate the presence of underlying inherited metabolic diseases. The present study was performed to investigate the presence of underlying metabolic diseases in patients with complex febrile seizures, using analyses of urine organic acids. METHODS: We retrospectively analyzed and compared the results of urine organic acid analysis with routine laboratory findings in 278 patients referred for complex febrile seizure. RESULTS: Of 278 patients, 132 had no abnormal laboratory findings, and 146 patients had at least one of the following abnormal laboratory findings: acidosis (n=58), hyperammonemia (n=55), hypoglycemia (n=21), ketosis (n=12). Twenty-six (19.7%) of the 132 patients with no abnormal findings and 104 (71.2%) of the 146 patients with statistically significant abnormalities showed abnormalities on the organic acid analysis (P<0.05). Mitochondrial respiratory chain disorders (n=23) were the most common diseases found in the normal routine laboratory group, followed by PDH deficiency (n=2 ) and ketolytic defect (n=1). In the abnormal routine laboratory group, mitochondrial respiratory chain disorder (n=29) was the most common disease, followed by ketolytic defects (n=27), PDH deficiency (n=9), glutaric aciduria type II (n=9), 3-methylglutaconic aciduria type III (n=6), biotinidase deficiency (n=5), propionic acidemia (n=4), methylmalonic acidemia (n=2), 3-hydroxyisobutyric aciduria (n=2), orotic aciduria (n=2), fatty acid oxidation disorders (n=2), 2-methylbranched chain acyl CoA dehydrogenase deficiency (n=2), 3-methylglutaconic aciduria type I (n=1), maple syrup urine disease (n=1), isovaleric acidemia (n=1), HMG-CoA lyase deficiency (n=1), L-2-hydroxyglutaric aciduria (n=1), and pyruvate carboxylase deficiency (n=1). CONCLUSION: These findings suggest that urine organic acid analysis should be performed in all patients with complex febrile seizure and other risk factors for early detection of inherited metabolic diseases.
Acetyl-CoA C-Acetyltransferase ; Acidosis ; Acyl-CoA Dehydrogenase ; Amino Acid Metabolism, Inborn Errors ; Biotinidase Deficiency ; Brain Diseases, Metabolic, Inborn ; Electron Transport ; Fever ; Humans ; Hydroxybutyrates ; Hyperammonemia ; Hypoglycemia ; Isovaleryl-CoA Dehydrogenase ; Ketosis ; Maple Syrup Urine Disease ; Metabolic Diseases ; Multiple Acyl Coenzyme A Dehydrogenase Deficiency ; Propionic Acidemia ; Pyruvate Carboxylase Deficiency Disease ; Pyruvate Dehydrogenase Complex Deficiency Disease ; Retrospective Studies ; Risk Factors ; Seizures ; Seizures, Febrile

Anemia, Hemolytic, Congenital Nonspherocytic ; Erythrocytes ; Humans ; Pyruvate Kinase/deficiency* ; Pyruvate Metabolism, Inborn Errors

OBJECTIVETo screen potential mutation and explore the underlying mechanism for a consanguineous pedigree featuring pyruvate kinase (PK) deficiency.

METHODSThe red blood cell pyruvate kinase activities of all family members were detected. All the exons and intron-exon boundaries of the PKLR gene for the proband were amplified and analyzed by direct sequencing. Restriction endonuclease enzymes were used to identify the presence of mutations of all family members.

RESULTSThe pyruvate kinase activities were 5.89 U/g Hb in the proband, 3.45, 6.54, 8.87, 7.89, 9.32 U/g Hb in his younger sister, father, mother, grandmother and elder aunt, respectively. The homozygous missense mutation of T>C transition at position 941 in exon 7 of PKLR gene resulted to a Ile314Thr substitution in the proband, and mutant alleles were identified at the level of RNA transcript by cDNA sequence analysis. His younger sister was also homozygous for Ile314Thr. Heterozygosity for Ile314Thr was confirmed in his grandmother, parents and elder aunt.

CONCLUSIONIle314Thr homozygous missense mutation in exon 7 of PKLR is the molecular mechanism of pyruvate kinase deficiency in this family.

Anemia, Hemolytic, Congenital Nonspherocytic ; genetics ; Child, Preschool ; Female ; Humans ; Male ; Pedigree ; Point Mutation ; Pyruvate Kinase ; deficiency ; genetics ; Pyruvate Metabolism, Inborn Errors ; genetics

OBJECTIVETo explore the molecular mechanism of erythrocyte pyruvate kinase deficiency (PKD).

METHODSTargeted sequence capture and next-generation sequencing (NGS) were used to detect the regions of exon and exon-intron boundarie of PKLR gene in a clinical suspected PKD patient. The protein function of mutant gene was forecasted by the SIFT and PolyPhen-2 databank, after the mutation of PKLR gene in the patient was detected by the NGS technology, its genotype was confirmed by Sanger sequencing.

RESULTSThe patient was found to have peculiar double heterozygous mutations: 661 G>A (Asp221Asn) of exon 5 and 1528 C>T (Arg510Ter) of exon 10, resulting in amino acid substitution Asp221Asn and Arg510Ter, these mutations were also further confirmed by Sanger sequencing. The complex mutations were infrequent and each of them was able to cause diseases.

CONCLUSIONThe complex mutations of both 661 G>A and 1528 C>T of PKLR gene are the molecular mechanism of PKD. Simultaneous existance of above-mentioned complex mutations in PDK patient was never been previously reported at home and abroad.

Anemia, Hemolytic, Congenital Nonspherocytic ; genetics ; Exons ; Genotype ; High-Throughput Nucleotide Sequencing ; Humans ; Introns ; Mutation ; Pyruvate Kinase ; deficiency ; genetics ; Pyruvate Metabolism, Inborn Errors ; genetics

OBJECTIVETo evaluate the feasibility of genetic and prenatal diagnosis for a family affected with pyruvate kinase deficiency (PKD).

METHODSTargeted sequence capture and high-throughput sequencing technology was used to detect the exons and exon-intron boundaries of the PKLR gene in a clinically suspected PKD patient. Meanwhile, the genotype of the pedigree was validated by Sanger sequencing. Prenatal genetic diagnosis was performed by amniotic fluid sampling after genotype of the mother of the proband was determined.

RESULTSThe proband was found to harbor double heterozygous mutations, c.661G>A (Asp221Asn) and c.1528C>T (Arg510Ter), which resulted in amino acid substitution Asp221Asn and Arg510Ter. Such mutations were confirmed by Sanger sequencing. The mother and father of the proband were detected to have respectively carried c.1528C>T (Arg510Ter) and c.661G>A (Asp221Asn) mutation. The fetus was found to have carried the same mutations as the proband. Following selected abortion, analysis of fetal tissue was consistent with the result of prenatal diagnosis.

CONCLUSIONThe compound mutations of c.661G>A and c.1528C>T of PKLR gene probably underlie the PKD in the family. Prenatal diagnosis of the mutations analysis can facilitate detection of affected fetus in time.

Adult ; Anemia, Hemolytic, Congenital Nonspherocytic ; embryology ; enzymology ; genetics ; Base Sequence ; Child, Preschool ; DNA Mutational Analysis ; Exons ; Female ; Genotype ; Humans ; Male ; Molecular Sequence Data ; Mutation ; Pedigree ; Pregnancy ; Prenatal Diagnosis ; Pyruvate Kinase ; deficiency ; genetics ; metabolism ; Pyruvate Metabolism, Inborn Errors ; embryology ; enzymology ; genetics

No abstract available.
Hypertension ; Leigh Disease*

Introduction: Leigh disease and Leigh-like syndrome are a heterogenous group of neurodegenerative disorders involving any level of the neuraxis and may present with a variety of clinical presentations, prominent among them is psychomotor regression. Despite the remarkable number of established disease genes and novel mutations being discovered, many cases of Leigh syndrome remain without a genetic diagnosis, indicating that there are still more disease genes to be identified. Case: Here we present a case of a two and a half-year-old girl who presented with delayed acquisition of developmental milestones with subsequent regression, ataxia, and dyskinesia. Her work-up showed raised blood lactate levels and lactate peak in MR spectroscopy. Mitochondria genome showed absence of mitochondrial DNA mutation, while whole exome sequence analysis revealed a novel dynein gene variant, p.A1577S. Her parents underwent genetic testing as well, and her father also had the same dynein mutation, however, is non-symptomatic. She had an older brother who initially presented with ophthalmoplegia and eventually developed psychomotor regression. He subsequently expired from respiratory failure after almost 2 years from initial presentation. Both siblings were diagnosed with Leigh syndrome. Conclusion The diagnosis of Leigh syndrome remains based on characteristic clinical and radiologic findings. However, a specific defect must be identified if reliable genetic counseling is to be provided.
Neurodegenerative Diseases|leigh Disease

Acute Disease ; Female ; Humans ; Infant ; Infant, Newborn ; Propionates ; blood ; Pyruvate Carboxylase Deficiency Disease

OBJECTIVE: To analyze the clinical features and genetic basis for a child with pyruvate carboxylase deficiency type A (PCD-A). METHODS: Clinical data of the child was retrospectively analyzed. The child and his parents were subjected to trio-whole exome sequencing, and candidate variants were verified by bioinformatics analysis. RESULTS: The child was admitted due to fever with vomiting and disturbance of consciousness. His clinical manifestations included severe decompensated acidosis, hypotension and intractable shock. Cranial MRI showed abnormal signal in the brain, and chest X-ray revealed acute pulmonary edema. DNA sequencing revealed that he has harbored compound heterozygous variants of the PC gene, namely c.182T>C (p.I61T) and c.2581G>A (p.V861M), which were respectively inherited from his father and mother. Neither variant was retrievable in the ClinVar and HGMD databases. Through prediction of protein structure, both variants may affect the functional stability of the protein product. CONCLUSION The compound heterozygous variants of the PC gene probably underlay the PCD-A in this child. Combined with the clinical features, the child was ultimately diagnosed as PCD-A. Above finding has enriched the spectrum of PC gene variation underlying PCD-A.
Child ; Family ; Humans ; Male ; Mutation ; Pyruvate Carboxylase Deficiency Disease ; Retrospective Studies ; Exome Sequencing

Central Nervous System ; pathology ; Humans ; Infant ; Leigh Disease ; pathology ; physiopathology