[Show abstract][Hide abstract]ABSTRACT: Background:
Early detection of critical congenital heart disease (CCHD) can significantly reduce morbidity and mortality among newborns. We investigate the feasibility of implementing a community-based newborn CCHD screening program in Taipei.
Twelve birthing facilities in Taipei participated in a trial screening program between October 1, 2013, and March 31, 2014. Newborns underwent pulse oximetry at 24-36 h old, with probes attached to the right hand and one lower limb. Any screening saturation ≥95% in either extremity, with an absolute difference of ≤3% between the right hand and foot, was accepted as a screening pass. A screening result was considered as a fail if the oxygen saturation was <95% at either probe site, on 3 separate occasions, each separated by 30 min or the first result was <95% at either probe site, and any subsequent oxygen saturation measurement was <90%. Public health nurses would follow up all missed or refused cases.
Of the 6,387 live births, 6,296 newborns (coverage rate: 6,296/6,387 = 98.6%) underwent appropriate pulse oximetry screening. Sixteen newborns (0.25%) were reported to have a failed screening result. Five of these screen positive newborns were confirmed with CCHD; two of them were diagnosed solely attributed to the failed screening results. The false-positive rate was 0.18%. Implementing a 6-month screening program for CCHD produced good case detection rate, while using efficient screening and referral systems.
This program was successful in integrating screening, referral and public health tracking systems. The protocol outlined in this report could provide a community-based model for worldwide implementation.
[Show abstract][Hide abstract]ABSTRACT: Background:
Propionyl-CoA carboxylase (PCC) is a mitochondrial enzyme involved in the catabolism of several essential amino acids and odd chain fatty acids. Previous PCC assays have involved either a radiometric assay or have required mitochondria isolation and/or enzyme purification.
We developed an enzymatic method to analyze PCC activity in phytohaemagglutinin (PHA) stimulated lymphocytes that involves high performance liquid chromatography.
The method shows good linearity and sensitivity. PCC activity was unaffected even when lymphocytes were isolated and PHA stimulated after a whole blood sample had been stored at 4°C for 5days. This indicates that this method is suitable for analyzing samples from distant medical centers. The PCC activity of patients with propionic acidemia was found to be much lower than that of normal individuals and carriers. However, this PCC assay is significantly affected by the red blood cell contamination. In conclusion, this is a reliable method for performing PCC assays and only requires 0.5 to 1.0ml of whole blood from newborns.
The PCC assay established in this study is useful for the confirmation of PA in individuals, and prenatal diagnosis and genetic counseling for the affected families.
No preview · Article · Nov 2015 · Clinica chimica acta; international journal of clinical chemistry
[Show abstract][Hide abstract]ABSTRACT: Propionyl-CoA carboxylase (PCC) is involved in the catabolism of branched chain amino acids, odd-numbered fatty acids, cholesterol, and other metabolites. PCC consists of two subunits, α and β, encoded by the PCCA and PCCB genes, respectively. Mutations in the PCCA or PCCB subunit gene may lead to propionic acidemia. In this study, we performed mutation analysis on ten propionic acidemia patients from eight unrelated and nonconsanguineous families in Taiwan. Two PCCA mutations, c.229C→T (p.R77W) and c.1262A→C (p.Q421P), were identified in a PCCA-deficient patient. Six mutations in the PCCB gene, including c.-4156_183+3713del, c.580T→C (p.S194P), c.838dup (p.L280Pfs*11), c.1301C→T (p.A434V), c.1316A→G (P.Y439C), and c.1534C→T (p.R512C), were identified in seven PCCB-deficient families. The c.-4156_183+3713del mutation is the first known large deletion that affects the PCCB gene functions. Furthermore, the c.1301C→T and c.-4156_183+3713del mutations in the PCCB gene have not been reported previously. Clinical features demonstrated that these two frequent mutations are associated with low enzyme activity and a classic propionic acidemia phenotype.
No preview · Article · May 2014 · Biochemical Genetics
[Show abstract][Hide abstract]ABSTRACT: 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 Ex6-96A>G, account for 23.2%, 12.0% and 9.2%, of the mutant alleles, respectively. Haplotype analysis shows that R241C and Ex6-96A>G 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 -4173_-407del, 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.Journal of Human Genetics advance online publication, 9 January 2014; doi:10.1038/jhg.2013.136.
No preview · Article · Jan 2014 · Journal of Human Genetics
[Show abstract][Hide abstract]ABSTRACT: To establish a hearing screening program with high coverage, low referral rate, high follow-up rate, and early intervention in Taipei City.
From September 2009 to December 2010, 85% delivery units in Taipei City, which includes 20 hospitals and 14 obstetrics clinics, were recruited into the screening program in two stages. A total of 15,930 babies were born in these participating hospitals and clinics during the program period. Among these neonates, 15,790 underwent hearing screening test with automatic auditory brainstem response (AABR). The screening was free of charge to the parents. The hearing screening examination was performed 24-36h after birth. The same test was repeated between 36 and 60h of age if the baby failed the first hearing test. The neonate was referred to the diagnostic hospitals for further investigations if he failed the second test.
The screening coverage rate was 99.1% (15,790/15,930). The incidence of bilateral moderate to severe and unilateral hearing loss was 1.4 per 1000 (22/15,790) and 1.5 per 1000 (24/15,790), respectively. Four percent (626/15,790) of newborns failed to pass the initial screening test and 1.0% of newborns failed to pass the second screening test. Therefore, 1.0% newborns were referred for diagnostic assessments. The follow-up rate was 94.4% (151/160). Sixty-four percent (14/22) of babies with bilateral hearing loss completed the full diagnostic hearing tests within 3 months of birth.
The universal newborn hearing screening program is an adequate program for Taipei City with high coverage, low referral rate, and good follow-up rate. Screening fees covered by third parties, two-stage screening steps with AABR strategy, and the stringent monitoring system proved to be effective.
2b, individual cohort study.
No preview · Article · Aug 2013 · International journal of pediatric otorhinolaryngology
[Show abstract][Hide abstract]ABSTRACT: Predicted protein antigenicity and continuous B-cell epitopes of identified lipoproteins from Mycoplasma fermentans M64. The data was sorted by the predicted antigenicity likelihood of the lipoproteins.
[Show abstract][Hide abstract]ABSTRACT: Mycoplasma fermentans is a potent human pathogen which has been implicated in several diseases. Notably, its lipid-associated membrane proteins (LAMPs) play a role in immunomodulation and development of infection-associated inflammatory diseases. However, the systematic protein identification of pathogenic M. fermentans has not been reported. From our recent sequencing results of M. fermentans M64 isolated from human respiratory tract, its genome is around 1.1 Mb and encodes 1050 predicted protein-coding genes. In the present study, soluble proteome of M. fermentans was resolved and analyzed using two-dimensional gel electrophoresis. In addition, Triton X-114 extraction was carried out to enrich amphiphilic proteins including putative lipoproteins and membrane proteins. Subsequent mass spectrometric analyses of these proteins had identified a total of 181 M. fermentans ORFs. Further bioinformatics analysis of these ORFs encoding proteins with known or so far unknown orthologues among bacteria revealed that a total of 131 proteins are homologous to known proteins, 11 proteins are conserved hypothetical proteins, and the remaining 39 proteins are likely M. fermentans-specific proteins. Moreover, Triton X-114-enriched fraction was shown to activate NF-kB activity of raw264.7 macrophage and a total of 21 lipoproteins with predicted signal peptide were identified therefrom. Together, our work provides the first proteome reference map of M. fermentans as well as several putative virulence-associated proteins as diagnostic markers or vaccine candidates for further functional study of this human pathogen.
[Show abstract][Hide abstract]ABSTRACT: COG functional profile of the 11 mycoplasmal species. The distribution of COG of total, identified and predicted-only proteins were shown as black, red and blue bars respectively. The organism names of human pathogens were labeled in red. A) COG classes related to Information Storage and Processing (J, K and L categories); B) and C) Cellular Processes (D, V, T, M, U and O categories); D) and E) Metabolism (C, G, E, F, H, I, P and Q categories) and F) Others (R, S and no COG assignment).
[Show abstract][Hide abstract]ABSTRACT: Multiple sequence alignment of the agalactiae-P80 and hominis-P80 mycoplasmal lipoproteins. The hominis-P80 cluster contains proteins similar to the M. hominis P80 and the agalactiae-P80 otherwise. Sequence conservation was shown in a gradient from blue (the most conserved), black to red (the least conserved).
[Show abstract][Hide abstract]ABSTRACT: Conservation of M. fermentans M64 proteins conserved across the fully sequenced Mycoplasma species. The values showed the number and percentage of identified and predicted M. fermentans M64 proteins that are homologous with species in each row. The two percentage columns were colored according to the level of conservation, ranging from dark red (highest conservation) to dark green (lowest conservation).
[Show abstract][Hide abstract]ABSTRACT: List of the M. fermentans M64 proteins identified in this study. The summary table includes the protein annotation, MASCOT scores, predicted pIs and molecular masses, COG classification, TMHMM and LipoP prediction results, KEGG orthology (KO) and pathway information.
[Show abstract][Hide abstract]ABSTRACT: The circular representation of the M. fermentans M64 genome. The ticks on the outermost concentric circle indicate the relative genomic positions in base pairs, where position one is the first base of the upstream intergenic region of dnaA gene. Second and third concentric circles: predicted protein coding genes on the plus and minus strands, respectively. The various functional groups of the predicted genes (color coded) are categorized according to the COG classification. Fourth concentric circle: locations of rRNA (gray) and tRNAs (green) genes. Fifth concentric circle: ICEF-I and IIs (red arrows), ICEF-III (green arrows), ΦMFV1 prophage (blue arrows), IS elements [IS1550 (dark brown), IS1630 (brown), ISMf1 (orange)], 16S-23S rRNA operons (blue), and non-IS repeats (yellow). Sixth and seventh concentric circles: GC content and GC skew calculated in 5,000 bp of sliding window with 100 bp of step window, respectively.
[Show abstract][Hide abstract]ABSTRACT: A global view of M. fermentans metabolic networks showing the connected reactions in carbohydrate (green), nucleotide (blue), amino acids (red), cofactors and vitamins (orange), and lipid (pink) metabolisms. The map is produced by matching the M64 predicted proteins to metabolic pathways with the “color pathway” tool in KEGG (http://www.genome.jp/kegg/tool/color_pathway.html).
[Show abstract][Hide abstract]ABSTRACT: Phylogenetic tree of the 21 Mycoplasma species analyzed in Figure 4. Maximum-likelihood method was performed on the 23S rRNA to reconstruct this tree using PHYML 3.0. The branch reliability was evaluated by implementing the aLRT method. Phytoplasma OY was set as the outgroup. The scale bar stands for the estimated number of nucleotide substitutions per site.
[Show abstract][Hide abstract]ABSTRACT: Comparison among ICEF-IA, ICEF-IIA, and ICEF-IIIA of M. fermentans M64 indicated ICEF-III is a new family of ICE.
A., B., and C. Results of sequence comparison of ICEF-IIIA and ICEF-IA, ICEF-IA and ICEF-IIA, and ICEF-IIA and ICEF-IIIA, respectively. The sequence similarities between ICEF-IIIA and ICEF-IA, ICEF-IA and ICEF-IIA, and ICEF-IIA and ICEF-IIIA are, 45.6%, 91.9%, and 45.4%, respectively. The GC content (sliding window: 120 bp) of each element was plotted on top or bottom of each panel. The arrows indicate the ORFs and the four-digit numbers represent the locus name (i.e., the number follow “MfeM64YM_” in locus tag). The two strips in dark gray stand for the forward and reverse strands of DNA. The direct and complementary matched regions between 2 elements are linked by blue and red lines, respectively. D. Phylogenetic tree of the 11 intact Integrative Conjugal Element (ICE) of Mycoplasmas. ICEB, ICEA, ICEF, ICEH, ICEM, and ICEC represent the elements in M. bovis, M. agalactiae, M. fermentans, M. hyopneumoniae, M. mycoides, and M. capricolum genomes, respectively. The scale bar stands for the estimated number of nucleotide substitutions per site.