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Objective: To determine whether patients with pyridoxine-responsive seizures but normal biomarkers for antiquitin deficiency and normal sequencing of the ALDH7A1 gene may have PNPO mutations. Methods:We sequenced the PNPO gene in 31 patients who fulfilled the above-mentioned criteria.

We report on a male proband with pyridoxine-dependent epilepsy (PDE) and neonatal seizure onset. At the age of 31 months, a prolonged status epilepticus led to severe neurological regression with cortical blindness, loss of speech and muscular hypotonia with slow recovery over the following 3 months. At 33 months of age pyridoxine therapy was initiated with excellent response and the boy remained seizure-free on pyridoxine monotherapy, except for two occasions with seizure recurrence 10 days after accidental pyridoxine withdrawal. alpha-aminoadipic semialdehyde dehydrogenase (antiquitin) deficiency was indicated by elevated pipecolic acid concentrations in plasma and alpha-aminoadipic semialdehyde excretion in urine. Molecular analysis of the antiquitin gene revealed a novel missense mutation c.57insA, while the mutation of the other allele remained unidentified so far. Despite the delay in diagnosis and prolonged status epilepticus, neuropsychological evaluations at the ages of 11 and 18 years demonstrated full-scale IQ of 93 and 92, respectively, with better verbal IQ (103 and 101) than performance IQ (85 and 82).

GM1 gangliosidosis manifests with progressive psychomotor deterioration and dysostosis of infantile, juvenile, or adult onset, caused by alterations in the structural gene coding for lysosomal acid beta-galactosidase (GLB1). In addition, allelic variants of this gene can result in Morquio B disease (MBD), a phenotype with dysostosis multiplex and entire lack of neurologic involvement. More than 100 sequence alterations in the GLB1 gene have been identified so far, but only few could be proven to be predictive for one of the GM1 gangliosidosis subtypes or MBD. We performed genotype analyses in 16 GM1 gangliosidosis patients of all phenotypes and detected 28 different genetic lesions. Among these, p.I55FfsX16, p.W65X, p.F107L, p.H112P, p.C127Y, p.W161X, p.I181K, p.C230R, p.W273X, p.R299VfsX5, p.A301V, p.F357L, p.K359KfsX23, p.L389P, p.D448V, p.D448GfsX8, and the intronic mutation IVS6-8A>G have not been published so far. Due to their occurrence in homozygous patients, four mutations could be correlated to a distinct GM1 gangliosidosis phenotype. Furthermore, the missense mutations from heteroallelic patients and three artificial nonsense mutations were characterized by overexpression in COS-1 cells, and the subcellular localization of the mutant proteins in fibroblasts was assessed. The phenotype specificity of 10 alleles can be proposed on the basis of our results and previous data.

Summary The diagnosis of Anderson–Fabry disease is often delayed or even missed. As severe renal manifestations are a hallmark of alfa‐galactosidase A (AGAL) deficiency, we tested the hypothesis that Anderson–Fabry disease is under‐recognized among male kidney transplant recipients. This nation‐wide study in Austria enrolled 1306 patients (ca 65% of all kidney transplanted males) from 30 kidney centers. AGAL activity was determined from filter paper dried blood spots by a fluorescence assay. A positive screening test was defined by an AGAL activity below 1.5 nmol/h/ml. In patients with a positive blood spot‐screening test, AGAL activity was re‐examined in peripheral blood leukocytes. Genetic testing for mutations in the GLA gene was performed by sequencing to confirm the diagnosis of Anderson–Fabry disease. Two previously not recognized cases with Anderson–Fabry disease were identified. Our study is the first showing that a diagnosis of Anderson–Fabry disease can be missed even in patients who undergo kidney transplantation. Case‐finding strategies may be considered a useful tool for diagnosis of this rare disease that may be somewhat more prevalent among kidney transplant recipients compared with dialysis populations.

Freshly harvested Florida pusley (Richardia scabraL.) seeds do not germinate in continuous dark, but do germinate if exposed to more than 2 hr of light each day. Increasing periods of illumination up to 16 hr per day increases the germination percentages. Scarification increases the rate of germination in the presence of light. Scarification also causes germination of seeds stored for 8 months or longer in total darkness. The seeds fail to germinate at constant temperatures of 15 C or less and at 40 C. Almost complete germination occurs at a constant temperature of 30 C, or at alternating temperatures of 20 and 30 C. The seeds germinate equally well in the pH range of 3 to 8. Increasing depths of planting reduces percent emergence, and none of the seedlings emerge from a depth of 1.5 cm or more. Air-dried seeds can be stored either at 5 or at 25 C without losing viability, at least up to a period of 1 year after harvest. Moist storage of seeds at 5 C reduces germinability.

Objective: To investigate the role of intragenic deletions of ALDH7A1 in patients with clinical and biochemical evidence of pyridoxine-dependent epilepsy but only a single identifiable mutation in ALDH7A1.Methods: We designed a custom oligonucleotide array with high-density probe coverage across the ALDH7A1 gene. We performed array comparative genomic hybridization in 6 patients with clinical and biochemical evidence of pyridoxine-dependent epilepsy but only a single detectable mutation in ALDH7A1 by sequence analysis. Results:We found partial deletions of ALDH7A1 in 5 of 6 patients. Breakpoint analysis reveals that the deletions are likely a result of Alu-Alu recombination in all cases. The density of Alu elements within introns of ALDH7A1 suggests susceptibility to recurrent rearrangement. Conclusion:Patients with clinical pyridoxine-dependent epilepsy and a single identifiable mutation in ALDH7A1 warrant further investigation for copy number changes involving the ALHD7A1 gene. Neurology ® 2015;85:756-762 GLOSSARY a-AASA 5 a-aminoadipic semialdehyde; CGH 5 comparative genomic hybridization; PDE 5 pyridoxine-dependent epilepsy.First described in 1954, pyridoxine-dependent epilepsy (PDE) is a metabolic epileptic encephalopathy characterized by pharmacoresistant seizures that typically come under control after initial administration followed by supplementation of pyridoxine at pharmacologic doses. The biochemical and genetic bases of this rare familial epilepsy were solved in 2006 when mutations in ALDH7A1 resulting in dysfunction of the protein antiquitin were discovered. 1 Metabolic changes consistent with PDE can be detected by measuring elevated levels of the biomarker a-aminoadipic semialdehyde (a-AASA) in various body fluids.1,2 As elevations of a-AASA are also present in patients with molybdenum cofactor deficiency and isolated sulfite oxidase deficiency, 3 genotyping of ALDH7A1 is required to confirm the diagnosis. In the vast majority of published cases, homozygous or compound heterozygous mutations of both ALDH7A1 alleles have been detected.We investigated 6 patients with a clinical diagnosis of PDE and positive biomarkers in which only a single, heterozygous mutation in ALDH7A1 could be identified by sequence analysis. We designed a custom oligonucleotide array that included high-density probe coverage of the ALDH7A1 gene to look for intragenic deletions or duplications that would have been missed by conventional sequence analysis. Using this strategy, we found partial deletions of ALDH7A1 in 5 of 6 patients, each of which is likely the result of an Alu-Alu recombination event. Our results suggest that in patients with clinical and biochemical evidence of PDE in the setting of a

Phenylketonuria (PKU, MIM 261600) is an autosomal recessive disorder caused by mutations of the phenylalanine hydroxylase gene (PAH, GenBank U49897.1, RefSeq NM_000277). To date more than 560 variants of the PAH gene have been identified. In Europe there is regional distribution of specific mutations. Due to recent progress in chaperone therapy, the prevalence of BH4-responsive alleles gained therapeutic importance. Here we report the mutational spectrum of PAH deficiency in 147 unrelated Austrian families. Overall mutation detection rate was 98.6 %. There was a total of 62 disease-causing mutations, including five novel mutations IVS4 + 6T>A, p.H290Y, IVS8-2A>G, p.A322V and p.I421S. The five most prevalent mutations found in patients were p.R408W, IVS12 + 1G>A, p.R261Q, p.R158Q and IVS2 + 5G>C. Neonatal phenylalanine levels before treatment were available in 114/147 patients. Prediction of BH4-responsiveness in patients with full genotypes was exclusively made according to published data. Among the 133 patients needing dietary treatment, 28.4 % are expected to be BH4 "non-responsive", 4.5 % are highly likely BH4-responsive, 35.8 % are probably BH4-responsive while no interpretation was possible for 31.3 %. The mutation data reflect the population history of Austria and provide information on the likely proportion of Austrian PKU patients that may benefit from BH4-therapy.