Epigenetic profiling of the H19 differentially methylated region and comprehensive whole genome array-based analysis in Silver-Russell syndrome.

Silver-Russell syndrome (SRS) is a clinically and genetically heterogeneous congenital disorder characterized by severe growth retardation. Hypomethylation of the differentially methylated region (DMR) of the H19 gene and uniparental disomy of maternal chromosome 7 is present in ∼45% of the patients with SRS so more than half of these patients have no known genetic etiology. We combined several molecular technologies including multiplex methylation polymerase chain reaction, methylation-sensitive multiple ligation probe-dependent amplification, and methylation-sensitive high-resolution melting to assess the epigenetic status of 34 patients with SRS. Additionally, we applied a whole genome strategy to detect copy number changes and loss of heterozygosity. Thirteen patients (38.2%) had hypomethylation of the DMR of the H19 gene and none had uniparental disomy of maternal chromosome 7. The whole genome arrays identified five patients (14.7%) with microdeletions on chromosomes 1q23q24.3, 7p15.3, 13q31.3, 14q32.31, and 15q26.2qter, respectively. The overall mutation detection rate was 52.9% by the epigenetic study and the whole genome strategy. Although epimutation may be the major cause of SRS and can be identified by multiplex methylation polymerase chain reaction, the whole genome approach also provides information on the etiology of SRS. If no epimutation is identified in the patients with typical SRS, microdeletions should be suspected.

Development and psychometric properties of the dialysis module of the WHOQOL-BREF Taiwan version.

BACKGROUND: Quality of life (QOL) is now considered to be an important part of the assessment of dialysis patients. The aim of this study was to develop and assess the reliability, validity and sensitivity of the dialysis module of the World Health Organization Quality of Life - Brief (WHOQOL-BREF) Taiwan version [WHOQOL-BREF(TW)] in patients undergoing regular hemodialysis (HD). METHODS: QOL survey was administered to 283 regular HD patients in metropolitan Taipei. The instruments used included: (1) the proposed module - composed of the core part, the WHOQOL-BREF(TW), and the six specific items; (2) the symptom/problem (S/P) scale - composed of 12 items specific for dialysis patients; (3) the utility measure, which was performed with standard gamble (SG) methods; and (4) the rating scale (RS). RESULTS: Based on the six criteria of validity, reliability and variance of the items, four HD-specific items were selected. Reliability study showed that Cronbach's alphas, composite reliability, and test-retest reliability (intraclass correlation at an average retest interval of 4-8 weeks) of the four domains of physical, psychological, social relationship and environment, ranged from 0.74-0.82, 0.79-0.84 and 0.61-0.79, respectively. Validity study showed that all the correlations between an item and its corresponding domain were highly significant (r>0.4, p<0.01) and larger than the correlations between the item and other domains. SG and psychometric measures showed relatively low correlations (0.12-0.26). The module showed the same construct as the WHOQOL-BREF(TW) under confirmatory factor analysis, whereas the exploratory factor analysis showed mild variation. Convergent and discriminant validity were good. Global QOL, physical, psychological and environment domains had some sensitivity to differentiate the severity of the condition of patients receiving HD. Clinical validity was demonstrated in global QOL, physical and psychological domains to have significant correlations with S/P scores. CONCLUSION: Besides broader coverage than the core WHOQOL-BREF(TW), the dialysis module of the WHOQOL-BREF(TW) is a valid, reliable and sensitive QOL instrument for the assessment of HD patients in Taiwan.

A novel DCX missense mutation in a family with X-linked lissencephaly and subcortical band heterotopia syndrome inherited from a low-level somatic mosaic mother: Genetic and functional studies.

PURPOSE: To study the genetics and functional alteration of a family with X-linked lissencephaly and subcortical band heterotopia. METHODS: Five affected patients (one male with lissencephaly, four female with subcortical band heterotopia) and their relatives were studied. Sanger sequencing of DCX gene, allele specific PCR and molecular inversion probe technique were performed. Mutant and wild type of the gene products, namely doublecortin, were expressed in cells followed by immunostaining to explore the localization of doublecortin and microtubules in cells. In vitro microtubule-binding protein spin-down assay was performed to quantify the binding ability of doublecortin to microtubules. KEY FINDINGS: We identified a novel DCX mutation c.785A > G, p.Asp262Gly that segregated with the affected members of the family. Allele specific PCR and molecular inversion probe technique demonstrated that the asymptomatic female carrier had an 8% mutant allele fraction in DNA derived from peripheral leukocytes. This mother had 7 children, 4 of whom were affected and all four affected siblings carried the mutation. Functional study showed that the mutant doublecortin protein had a significant reduction of its ability to bind microtubules. SIGNIFICANCE: Low level mosaicism could be a cause of inherited risk from asymptomatic parents for DCX related lissencephaly-subcortical band heterotopia spectrum. This is particularly important in terms of genetic counselling for recurrent risk of future pregnancies. The reduced binding affinity of mutant doublecortin may contribute to developmental malformation of the cerebral cortex.

PRRT2 mutations lead to neuronal dysfunction and neurodevelopmental defects.

Mutations in the proline-rich transmembrane protein 2 (PRRT2) gene cause a wide spectrum of neurological diseases, ranging from paroxysmal kinesigenic dyskinesia (PKD) to mental retardation and epilepsy. Previously, seven PKD-related PRRT2 heterozygous mutations were identified in the Taiwanese population: P91QfsX, E199X, S202HfsX, R217PfsX, R217EfsX, R240X and R308C. This study aimed to investigate the disease-causing mechanisms of these PRRT2 mutations. We first documented that Prrt2 was localized at the pre- and post-synaptic membranes with a close spatial association with SNAP25 by synaptic membrane fractionation and immunostaining of the rat neurons. Our results then revealed that the six truncating Prrt2 mutants were accumulated in the cytoplasm and thus failed to target to the cell membrane; the R308C missense mutant had significantly reduced protein expression, suggesting loss-of function effects generated by these mutations. Using in utero electroporation of shRNA into cortical neurons, we further found that knocking down Prrt2 expression in vivo resulted in a delay in neuronal migration during embryonic development and a marked decrease in synaptic density after birth. These pathologic effects and novel disease-causing mechanisms may contribute to the severe clinical symptoms in PRRT2-related diseases.

Quality of life and its determinants of hemodialysis patients in Taiwan measured with WHOQOL-BREF(TW).

BACKGROUND: In 1991, the World Health Organization (WHO) initiated a cross-cultural project to develop a quality-of-life (QOL) questionnaire (WHOQOL); soon after this, the clinically applicable short form was developed and named WHOQOL-BREF, followed by a Taiwanese version (WHOQOL-BREF[TW]). METHODS: We first administered the WHOQOL-BREF(TW) and symptom/problem scale to 376 patients with end-stage renal disease on regular hemodialysis therapy in Taiwan. Analysis with multiple stepwise regressions was conducted to study determinants of QOL domains and items. RESULTS: The WHOQOL-BREF(TW) was reliable and valid from various validation studies. The 4 domains (physical, psychological, social relations, and environment) and global items (overall quality of life and general health) of the WHOQOL-BREF(TW) each differentiated symptoms/problems of hemodialysis patients from age-, sex-, and education-matched healthy referents. The 4 domains, except for environment and global items of the WHOQOL-BREF(TW), each differentiated erythropoietin dosage from age-, sex-, and education-matched healthy referents. After adjusting for age, sex, marriage, and education, the prominent associated factors of various QOL domains and items were age, area (Taipei or Keelung), hemoglobin level, normalized protein catabolic rate, and symptom/problem scale. CONCLUSION: The WHOQOL-BREF(TW) is reliable and valid for long-term study of hemodialysis patients, and hemodialysis had negative impacts on QOL, especially in patients with more severe disease with greater symptom/problem scores, lower hemoglobin levels, and lower normalized protein catabolic rates.