Papers by Kwang-Jen Hsiao
Journal of Human Genetics, Jan 9, 2014
Phenylalanine hydroxylase (PAH) deficiency is responsible for most cases of phenylketonuria (PKU)... more Phenylalanine hydroxylase (PAH) deficiency is responsible for most cases of phenylketonuria (PKU). In this study of the PAH mutation spectrum in the Taiwanese population, 139 alleles were identified including 34 different mutations. The V190G, Q267R and F392I mutations are first reported in this study. The most common mutations, R241C, R408Q and Ex6-96A4G, account for 23.2%, 12.0% and 9.2%, of the mutant alleles, respectively. Haplotype analysis shows that R241C and Ex6-96A4G are exclusively associated with haplotype 4.3 to suggest founder effects. On the other hand, R408Q is found on two distinct haplotypes suggesting recurrent mutations. The spectrum of PAH mutations in Taiwan shows various links to those of other Asian regions, yet remarkable differences exist. Notably, R408Q, E286K and À4173_ À407del, accounting for 21% of all mutant alleles in Taiwan, are very rare or are undetected among PKU cohorts of other Asian regions to suggest local founder effects. Moreover, the low homozygosity value of 0.092 hints at a high degree of ethnic heterogeneity within the Taiwanese population. Our study of PAH mutation spectrum and the associated haplotypes is useful for subsequent study on the origin and migration pattern via Taiwan, an island at the historical crossroad of migration of ancient populations.
PubMed, 2003
To establish the neonatal screening method of glucose-6 phosphate dehydrogenase (G6PD) deficiency... more To establish the neonatal screening method of glucose-6 phosphate dehydrogenase (G6PD) deficiency, G6PD activity was measured using the fluorescence spot test (FST) using dried blood samples on filter paper. The G6PD/6PGD rate test of venous blood samples was further performed for confirmation. The positive G6PD deficiency rate was 4.2% and its detection rates were 3.7% for all neonates and 5.2% only for male newborns when FST was used for neonatal screening. Conformation rates by use of G6PD/ 6PGD ratio test for G6PD deficiency were 86.8% and 100% particularly in the severely deficient groups. Both sensitivity and specificity were very high in the severely deficient groups. FST can be used in neonatal screening of G6PD deficiency because of its high accuracy, applicability, and simplicity. Moreover, a high volume of dried blood samples on filter paper can be tested quickly. It is very favorable to diagnose and treat G6PD deficiency early in high incidence districts.
Jinrui idengaku zasshi, Mar 1, 1988
For the purpose of exploring the possibility of implementing a nationwide screening program for i... more For the purpose of exploring the possibility of implementing a nationwide screening program for inherited metabolic diseases and congenital hypothyroidism in Chinese newborn infants, a pilot study was initiated in 1983 to detect patients with phenylketonuria (PKU), galactosemia, homocystinuria, maple syrup urine disease and congenital hypothyroidism (CHT) in mildly mentally retarded (mostly IQ 50-75) school children in Taiwan. Of 4,744 blood samples collected on filter paper from 246 primary and junior high schools all over the island, preliminary screening disclosed six suspected positive cases of PKU and nine of CHT. Two cases of classical PKU, one case of atypical PKU caused by tetrahydrobiopterin deficiency, and seven cases of CHT (one hypoplastic, two athyroid, three ectopic and one dyshormonogenesis) were finally confirmed. In addition to the seven cases of CHT, one more case found by questionnaire survey. This case was missed from the screening because he was recognized as CHT previously and was on thyroxine replacement therapy during screening. The incidence is I/1,581 for PKU and 1/593 for CHT in these children.
PubMed, Oct 1, 1993
The study subjects were those over 30 years of age in Kin-Hu, Kinmen. From January to February 19... more The study subjects were those over 30 years of age in Kin-Hu, Kinmen. From January to February 1991, demographic data, physical data, life style and blood chemistry parameters were collected from 2929 respondents (1368 men and 1561 women) out of 4475 eligible subjects. The study, conducted by the Yang-Ming Crusade, used a structured questionnaire and venipuncture technique. After clarifying missing values, a total number of 2868 (64.1%; 1334 men and 1534 women) subjects had completed the data set. Of the total subjects, 32.3% were 30-39 years of age, 24.3% were 40-49 yrs, 22.4% were 50-59 yrs, and 21.0% were above 60 years old. Fifty-two percent of men and 2.8% of women, were current smokers. Most of the surveyed subjects described themselves as having infrequent physical activity (men vs. women, 66.4% vs. 81.0%, p < 0.001). Men had significantly lower body mass index (BMI) and higher waist-hip ratio (W/H) than women (BMI: 22.9 +/- 3.2 vs. 23.2 +/- 3.7, p < 0.01; W/H: 0.89 +/- 0.06 vs. 0.84 +/- 0.07, p < 0.01). Men also had higher fasting plasma glucose (FPG), systolic/diastolic blood pressure (SBP/DBP) than women (FPG: 99 +/- 25.3 vs. 95.7 +/- 25.3 mg/dl, p < 0.0001; SBP: 132.1 +/- 19 vs. 126.4 +/- 21.5 mmHg, p < 0.0001; DBP: 80.4 +/- 12.5 vs. 76.5 +/- 12.3 mmHg, p < 0.0001).(ABSTRACT TRUNCATED AT 250 WORDS)
Human Genetics, Feb 5, 2002
The liver-specific phenylalanine hydroxylase catalyzes the conversion of phenylalanine to tyrosin... more The liver-specific phenylalanine hydroxylase catalyzes the conversion of phenylalanine to tyrosine. Genetic defects in the gene result in the autosomal recessive disorder phenylketonuria. We have identified a phenylalanine hydroxylase mutation, designated as -4173_-407del, in a hyperphenylalaninemic patient with a 3.7-kb deletion in the 5&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;#39;-flanking region of the phenylalanine hydroxylase gene. Characterization of the deleted sequence has led to the identification of a novel liver-specific DNase I hypersensitive site located 3.3 kb upstream of the RNA initiation site of the phenylalanine hydroxylase gene. We show that this site comprises a liver-specific enhancer with cAMP responsiveness. We further show by mutational analysis that the enhancer carries a major hepatocyte nuclear factor-1-binding site important for the enhancer function but not for cAMP responsiveness. In transient transfection assays with a reporter gene, we demonstrate that a phenylalanine hydroxylase plasmid construct carrying the -4173_-407del mutation is severely impaired in phenylalanine hydroxylase transcriptional activity. Our data indicate that the 3.7-kb deletion uncovered in the genomic DNA of the phenylketonuria proband is linked to the observed phenylketonuria phenotype as a result of a deletion of the newly identified liver-specific enhancer. Our systematic approach to the analysis and subsequent discovery of the novel deletion mutation may be applicable to the search for other mutations in the 5&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;#39; regulatory region of phenylalanine hydroxylase and other genes.
Human Genetics, May 1, 1994
Fragile X syndrome is a genetic disorder caused by abnormal function of the FMR-1 gene. The major... more Fragile X syndrome is a genetic disorder caused by abnormal function of the FMR-1 gene. The majority of fragile X syndrome patients carry an expansion of the CGG tri-nucleotide repeat in the FMR-1 gene, whereas others have a deletion or a point mutation in the FMR-1 structural gene. In this report, we analyzed a typical family with three male patients. RNA from Epstein-Barr virus transformed lymphoblastoid cells was used for RNase protection assay and reverse transcription-polymerase chain reaction (RT-PCR) analysis. Five normal individuals and one asymptomatic heterozygote from this family expressed detectable FMR-1 transcripts, whereas three fragile X patients showed no sign of expression with either assay. To extend the application of this PCR-based assay to laboratory diagnosis of fragile X syndrome, we confirmed that dried blood samples collected on screening filter papers for newborns are an adequate source of RNA for RT-PCR. Moreover, fragile X patients from the study family and another family were reliably identified by the absence of the FMR-1-specific PCR product from the dried blood specimens. Our studies indicate that this simple assay can be used to diagnose the fragile X syndrome for the majority of male patients.
Journal of Paediatrics and Child Health, Aug 1, 1999
: We report a case of galactose‐1‐phosphate uridyl transferase (GALT) deficiency in a full‐term C... more : We report a case of galactose‐1‐phosphate uridyl transferase (GALT) deficiency in a full‐term Chinese neonate, who presented with atypical biochemical features of hyperammonaemia in addition to the classical presenting features of jaundice and lethargy after feeding. Red cell GALT activity was virtually absent in the patient while 50% of normal activity was found in parents and a sibling. Mutation screening excluded both Q188R and N314D as the causative mutation in GALT gene, which suggested a possible genetic segregation among ethnic groups. Data from a Taiwan screening program suggested that the incidence of the disease was approximately 1 in 400 000 in the Chinese population which was a sixth of that in Caucasian populations.
Pediatrics
OBJECTIVESAn extended newborn critical congenital heart disease (CCHD) screening program using ox... more OBJECTIVESAn extended newborn critical congenital heart disease (CCHD) screening program using oximetry has been implemented in Taipei, Taiwan since April 2014. This study was conducted to investigate the test accuracy and efficiency of this screening protocol.METHODSThis study analyzed data from 30 birthing facilities representing 87.9% of live births in Taipei. Positive screening was defined as oxygen saturation <95% in either extremity or a preductal-postductal oxygen saturation difference of >3%. This study cohort was used to retrospectively estimate outcomes on the basis of different CCHD screening protocols.RESULTSDuring the study period, 93 058 of 94 204 (98.8%) infants who had no prenatal suspicion were screened. The referral rate was 0.17% (156/93 058), and up to 90% of test-positive infants were referred within 48 hours of life. Forty-two CCHD cases without prenatal suspicion were detected and 97.6% were diagnosed within 72 hours of life. Of the screened newborns, 4 ...
Background. In 2003, Taiwan experienced a series of outbreaks of severe acute respiratory syndrom... more Background. In 2003, Taiwan experienced a series of outbreaks of severe acute respiratory syndrome (SARS) and 1 laboratory-contamination accident. Here we describe a new phylogenetic analytical method to study the sources and dissemination paths of SARS-associated coronavirus (SARS-CoV) infections in Taiwan. Methods. A phylogenetic analytical tool for combining nucleotide sequences from 6 variable regions of a SARS-CoV genome was developed by use of 20 published SARS-CoV sequences; and this method was validated by use of 80 published SARS-CoV sequences. Subsequently, this new tool was applied to provide a better understanding of the entire complement of Taiwanese SARS-CoV isolates, including 20 previously published and 19 identified in this study. The epidemiological data were integrated with the results from the phylogenetic tree and from the nucleotide-signature pattern. Results. The topologies of phylogenetic trees generated by the new and the conventional strategies were similar...
Journal of Biological Chemistry, 1978
The effects of ATP and divalent cations on a divalent cation-independent phosphorylase phosphatas... more The effects of ATP and divalent cations on a divalent cation-independent phosphorylase phosphatase of Mr = 35,000 (phosphatase S) purified from canine cardiac muscle have been studied. The enzyme can be rapidly inactivated by ATP or other nucleoside di- and triphosphates and PPi, but not by AMP, adenosine, adenine, Pi, EDTA, ethylene glycol bis(beta-aminoethyl ether)N,N&amp;amp;#39; -tetraacetic acid, 1,10-phenanthroline, or 8-hydroxyquinoline. After removing the inactivating agent, such as ATP or PPi, by gel filtraiton followed by exhaustive dialysis, the inactivated enzyme (apophosphatase S) can be reactivated by preincubating with Mn2+ or Co2+, but not with Mg2+, Ca2+, Ni2+, Zn2+, Fe2+, Cu2+, Ba2+, Hg2+, Pb2+, or Cd2+. The Mn2+ -reactivated enzyme, which is less active than the Co2+ -reactivated enzyme, can be again inactivated by preincubating with ATP. The present findings indicate that phosphatase S contains a tightly bound divalent cation, probably Mn2+, in the active site. ATP and PPi, due to their structural similarity to the phosphoprotein substrate and their ability to chelate metal ions, can readily enter the active site to remove the divalent cation(s) essential for the catalytic function. The present findings also indicate that phosphatase S, a common catalytic subunit of several larger molecular forms of nospecific phosphoprotein phosphatase in cardiac muscle, can exist in two interconvertible forms, a metallized form (active) and a demetallized form (inactive). ATP and metal ions may regulate this class of isozymes by mediating the interconversions.
Clinical Chemistry, 1986
This is a method for measuring alpha-fetoprotein (AFP) in eluates of dried-blood samples on filte... more This is a method for measuring alpha-fetoprotein (AFP) in eluates of dried-blood samples on filter paper by use of a simple, sensitive two-site enzyme immunoassay. The spot, 6 mm in diameter (equivalent to about 12 microL of whole blood), is incubated overnight with alkaline phosphatase conjugated to rabbit anti-AFP antibody in a tube containing a polystyrene bead coated with mouse monoclonal antibody to AFP. After the beads are washed the enzyme activities associated with them are determined colorimetrically, with p-nitrophenyl phosphate as substrate. The measurable range of AFP is from 9 to 900 micrograms per liter of plasma. AFP in the dried-blood spot as determined by this method correlated well with the AFP value for serum from the same blood sample as determined by radioimmunoassay (r = 0.957, p less than 0.001). Preliminary studies in which we used this method with 242 healthy blood donors and 60 patients with hepatocellular carcinoma indicate that it may be suitable for use ...
Zhonghua er ke za zhi. Chinese journal of pediatrics, 2003
To determine distribution and mutation pattern of type P ATP7B gene mutation and to explore genot... more To determine distribution and mutation pattern of type P ATP7B gene mutation and to explore genotype and phenotype correlation in patients with Wilson's disease (WD). Sixty patients with WD from 57 no kinship families, 37 male and 23 female, were enrolled in this study. The age of onset ranged from 4.6 - 39 years, < or = 16 years in 55 patients. Some exons of ATP7B gene mutation were analyzed in patients with WD by using biochemical methods, polymerase chain reaction-single strand configuration polymorphism (PCR-SSCP), restriction fragment and DNA sequence analysis. Totally 778 coding regions were identified with restriction enzyme Msp I. The activity of Cu-ATPase was assessed by measuring inorganic phosphorus in 3 patients with known genotype. Fifty-two of 60 patients (86%) had presented with hepatic manifestations, 30 of them had only hepatic manifestations, 12/52 patients had hepatic and neurological manifestations at the same time; 10/52 patients had hepatic and other sym...
Taiwan yi xue hui za zhi. Journal of the Formosan Medical Association, 1989
Five-hundred and fifty one mentally retarded children from seven institutes in Northern Taiwan we... more Five-hundred and fifty one mentally retarded children from seven institutes in Northern Taiwan were screened by dried blood spot for the detection of treatable congenital metabolic diseases, including congenital hypothyroidism, phenylketonuria, homocystinuria, maple syrup urine disease and galactosemia. We found 2 children (0.36%) with congenital hypothyroidism, 1 case (0.18%) of classical phenylketonuria and two cases (0.36%) of trisomy 21 associated with autoimmune thyroiditis. The results of our investigation suggest that congenital hypothyroidism and phenylketonuria can be the factors causing mental retardation among children in Taiwan and mass neonatal screening of these treatable inborn metabolic diseases is strongly indicated for efficiently circumventing mental retardation in our community.
Journal of human genetics, 2002
Congenital nephrogenic diabetes insipidus (NDI) is, in most instances, a rare X-linked recessive ... more Congenital nephrogenic diabetes insipidus (NDI) is, in most instances, a rare X-linked recessive renal disorder (MIM 304800) characterized by the clinical symptoms of polyuria, polydipsia, and dehydration. The X-linked NDI is associated with mutations of the arginine vasopressin receptor type 2 (AVPR2) gene, which results in resistance to the antidiuretic action of arginine vasopressin (AVP) in the renal tubules and collecting ducts. Identification of mutations in the AVPR2 gene can facilitate early diagnosis of NDI, which can prevent serious complications such as growth retardation and mental retardation. We analyzed three unrelated Chinese NDI families and identified three mutations: R106C, F287L, and R337X. In addition, an A/G polymorphism at cDNA nucleotide position 927 (codon 309L) was identified. A functional expression assay of the R106C and F287L mutants in COS-7 cells revealed that both mutants show significant dysfunction and accumulate intracellular cyclic adenosine monop...
Nephrology Dialysis Transplantation, 2005
Nature, 2005
The map-based sequence of the rice genome International Rice Genome Sequencing Project* Rice, one... more The map-based sequence of the rice genome International Rice Genome Sequencing Project* Rice, one of the world's most important food plants, has important syntenic relationships with the other cereal species and is a model plant for the grasses. Here we present a map-based, finished quality sequence that covers 95% of the 389 Mb genome, including virtually all of the euchromatin and two complete centromeres. A total of 37,544 nontransposable-element-related protein-coding genes were identified, of which 71% had a putative homologue in Arabidopsis. In a reciprocal analysis, 90% of the Arabidopsis proteins had a putative homologue in the predicted rice proteome. Twenty-nine per cent of the 37,544 predicted genes appear in clustered gene families. The number and classes of transposable elements found in the rice genome are consistent with the expansion of syntenic regions in the maize and sorghum genomes. We find evidence for widespread and recurrent gene transfer from the organelles to the nuclear chromosomes. The map-based sequence has proven useful for the identification of genes underlying agronomic traits. The additional single-nucleotide polymorphisms and simple sequence repeats identified in our study should accelerate improvements in rice production.
The Journal of Infectious Diseases, 2005
Background In 2003, Taiwan experienced a series of outbreaks of severe acute respiratory syndrome... more Background In 2003, Taiwan experienced a series of outbreaks of severe acute respiratory syndrome (SARS) and 1 laboratory-contamination accident. Here we describe a new phylogenetic analytical method to study the sources and dissemination paths of SARS-associated coronavirus (SARS-CoV) infections in Taiwan Methods A phylogenetic analytical tool for combining nucleotide sequences from 6 variable regions of a SARS-CoV genome was developed by use of 20 published SARS-CoV sequences; and this method was validated by use of 80 published SARS-CoV sequences. Subsequently, this new tool was applied to provide a better understanding of the entire complement of Taiwanese SARS-CoV isolates, including 20 previously published and 19 identified in this study. The epidemiological data were integrated with the results from the phylogenetic tree and from the nucleotide-signature pattern Results The topologies of phylogenetic trees generated by the new and the conventional strategies were similar, wit...
Journal of Human Genetics, 2005
Deficiency of citrin, a liver-type mitochondrial aspartate-glutamate carrier (AGC), encoded by th... more Deficiency of citrin, a liver-type mitochondrial aspartate-glutamate carrier (AGC), encoded by the SLC25A13 gene on chromosome 7q21.3, causes autosomal recessive disorders: adult-onset type II citrullinemia (CTLN2) and neonatal hepatitis associated with intrahepatic cholestasis (NICCD). So far, we have described 12 SLC25A13 mutations: 11 were from Japan and one from Israel. Three mutations found in Chinese and Vietnamese patients were the same as those in Japanese patients. In the present study, we identified a novel mutation IVS6+1G>C in a Japanese CTLN2 patient and widely screened 12 SLC25A13 mutations found in Japanese patients in control individuals from East Asia to confirm our preliminary results that the carrier frequency was high in Asian populations. Mutations 851-854del and 1638-1660dup were found in all Asian countries tested, and 851-854del associated with 290-haplotype in microsatellite marker D7S1812 was especially frequent. Other mutations frequently detected were IVS11+1G>A in Japanese and Korean, S225X in Japanese, and IVS6+5G>A in Chinese populations. We found a remarkable difference in carrier rates in China (including Taiwan) between north (1/ 940) and south (1/48) of the Yangtze River. We detected many carriers in Chinese (64/4169 = 1/65), Japanese (20/1372 = 1/69) and Korean (22/2455 = 1/112) populations, suggesting that over 80,000 East Asians are homozygotes with two mutated SLC25A13 alleles. Keywords Adult-onset type II citrullinemia (CTLN2) AE Aspartate-glutamate carrier (AGC) AE Citrin AE Carrier frequency AE SLC25A13 AE D7S1812
Human Mutation, 2001
The enzyme 6-Pyruvoyl-tetrahydropterin synthase (PTS) deficiency is the major cause of BH 4-defic... more The enzyme 6-Pyruvoyl-tetrahydropterin synthase (PTS) deficiency is the major cause of BH 4-deficient HPA. The frequency of BH 4-deficient HPA was estimated to be around 30% among Chinese HPA population in Taiwan, which is much higher than that in Caucasian population (1.5-2% of HPA). Approximately 86% of Chinese BH 4-deficient HPA was found to be caused by PTS-deficiency. Seven mutations-namely R25G, N52S, V56M, V70D, P87S, D96N, and T106M-had been identified in Chinese PTS-deficient patients previously. In this study, five additional mutations in the PTS gene, namely 200C>T (T67M), 226C>T (L76F), IVS3+1G>A (K54X), 116-119delTGTT (K38X) and 169-171delGTG (V57del), were identified by PCR and DNA sequencing in Chinese PTS-deficient patients. The 116-119delTGTT introduces a frameshift stop after lysine of codon 38 (K38X). The G-to-A transition at the consensus sequence of splicing donor site of exon 3 (IVS3+1G>A) resulted in exon 3 skipping of the PTS transcript and caused a frameshift stop after lysine of codon 54 (K54X). The T67M and V57del mutations have been found in Caucasian PTS deficient patients, while the L76F, IVS3+1G>A, and K38X mutations are novel. None of 100 normal alleles screened was found to have the L76F substitution, which indicated that the L76F substitution is a mutation causing PTS deficiency.
Uploads
Papers by Kwang-Jen Hsiao