Taiwan National Newborn Screening Program by Tandem Mass Spectrometry for Mucopolysaccharidoses Types I, II, and VI.

OBJECTIVE: To evaluate the initial cutoff values, rates of screen positives, and genotypes for the large-scale newborn screening program for multiple mucopolysaccharidoses (MPS) in Taiwan. STUDY DESIGN: More than 100 000 dried blood spots were collected consecutively as part of the national Taiwan newborn screening programs. Enzyme activities were measured by tandem mass spectrometry from dried blood spot punches. Genotypes were obtained when a second newborn screening specimen again had a decreased enzyme activity. Additional clinical evaluation was then initiated based on enzyme activity and/or genotype. RESULTS: Molecular genetic analysis for cases with low enzyme activity revealed 5 newborns with pathogenic alpha-L-iduronidase mutations, 3 newborns with pathogenic iduronate-2-sulfatase mutations, and 1 newborn was a carrier of an arylsulfatase B mutation. Several variants of unknown pathogenic significance were also identified, most likely causing pseudodeficiency. CONCLUSIONS: The highly robust tandem mass spectrometry-based enzyme assays for MPS-I, MPS-II, and MPS-VI allow for high-throughput newborn screening for these lysosomal storage disorders. Optimized cutoff values combined with second tier testing could largely eliminate false-positive results. Accordingly, newborn screening for these lysosomal storage disorders is possible.

Additional feeding during late gestation improves initial litter weight of lactating sows exposed to high ambient temperature

The present study investigated the impacts of additional feeding (AF; containing 16% excess feed than control diet in the last two weeks of gestation) during late gestation period and different backfat thickness (BFT) including low (<20 mm) and high (≥20 mm) backfat on reproductive performance, litter size, blood metabolites, hormonal profiles, colostrum, and milk composition of sows from day 90 of gestation to farrowing, and from farrowing to weaning during summer (average 28.3 °C). Fifty-four crossbred sows (Yorkshire × Landrace) were allotted to one of four treatments according to a 2×2 factorial arrangement. There was no AF×BFT interaction for any of the measured variables. The body weight change of sows was decreased for <20 mm in BFT and AF treatments during late gestation and farrowing to weaning periods. The BFT change of sows was higher in AF during late gestation and higher in ≥20 mm BFT during late gestation and farrowing to weaning periods. There were no effects of AF and BFT on the average daily feed intake and weaning-to-oestrus interval; however, sows in AF groups had a greater litter uniformity at birth and initial litter weight. Hormone profiles, colostrum, and milk composition of sows were not different among the treatments. Additional feeding during the last two weeks of gestation increases initial litter size in summer; however, final litter weight is not affected.(AU)

Isolation and characterization of a novel endo-beta-1, 4-glucanase from a metagenomic library of the black-goat rumen

ABSTRACT The various types of lignocellulosic biomass found in plants comprise the most abundant renewable bioresources on Earth. In this study, the ruminal microbial ecosystem of black goats was explored because of their strong ability to digest lignocellulosic forage. A metagenomic fosmid library containing 115,200 clones was prepared from the black-goat rumen and screened for a novel cellulolytic enzyme. The KG35 gene, containing a novel glycosyl hydrolase family 5 cellulase domain, was isolated and functionally characterized. The novel glycosyl hydrolase family 5 cellulase gene is composed of a 963-bp open reading frame encoding a protein of 320 amino acid residues (35.1 kDa). The deduced amino acid sequence showed the highest sequence identity (58%) for sequences from the glycosyl hydrolase family 5 cellulases. The novel glycosyl hydrolase family 5 cellulase gene was overexpressed in Escherichia coli. Substrate specificity analysis revealed that this recombinant glycosyl hydrolase family 5 cellulase functions as an endo-β-1,4-glucanase. The recombinant KG35 endo-β-1,4-glucanase showed optimal activity within the range of 30-50 °C at a pH of 6-7. The thermostability was retained and the pH was stable in the range of 30-50 °C at a pH of 5-7.

Isolation and characterization of a novel endo--1, 4-glucanase from a metagenomic library of the black-goat rumen

ABSTRACT The various types of lignocellulosic biomass found in plants comprise the most abundant renewable bioresources on Earth. In this study, the ruminal microbial ecosystem of black goats was explored because of their strong ability to digest lignocellulosic forage. A metagenomic fosmid library containing 115,200 clones was prepared from the black-goat rumen and screened for a novel cellulolytic enzyme. The KG35 gene, containing a novel glycosyl hydrolase family 5 cellulase domain, was isolated and functionally characterized. The novel glycosyl hydrolase family 5 cellulase gene is composed of a 963-bp open reading frame encoding a protein of 320 amino acid residues (35.1 kDa). The deduced amino acid sequence showed the highest sequence identity (58%) for sequences from the glycosyl hydrolase family 5 cellulases. The novel glycosyl hydrolase family 5 cellulase gene was overexpressed in Escherichia coli. Substrate specificity analysis revealed that this recombinant glycosyl hydrolase family 5 cellulase functions as an endo-β-1,4-glucanase. The recombinant KG35 endo-β-1,4-glucanase showed optimal activity within the range of 30-50 °C at a pH of 6-7. The thermostability was retained and the pH was stable in the range of 30-50 °C at a pH of 5-7.(AU)

Isolation and characterization of a novel endo-ß-1,4-glucanase from a metagenomic library of the black-goat rumen.

The various types of lignocellulosic biomass found in plants comprise the most abundant renewable bioresources on Earth. In this study, the ruminal microbial ecosystem of black goats was explored because of their strong ability to digest lignocellulosic forage. A metagenomic fosmid library containing 115,200 clones was prepared from the black-goat rumen and screened for a novel cellulolytic enzyme. The KG35 gene, containing a novel glycosyl hydrolase family 5 cellulase domain, was isolated and functionally characterized. The novel glycosyl hydrolase family 5 cellulase gene is composed of a 963-bp open reading frame encoding a protein of 320 amino acid residues (35.1kDa). The deduced amino acid sequence showed the highest sequence identity (58%) for sequences from the glycosyl hydrolase family 5 cellulases. The novel glycosyl hydrolase family 5 cellulase gene was overexpressed in Escherichia coli. Substrate specificity analysis revealed that this recombinant glycosyl hydrolase family 5 cellulase functions as an endo-ß-1,4-glucanase. The recombinant KG35 endo-ß-1,4-glucanase showed optimal activity within the range of 30-50°C at a pH of 6-7. The thermostability was retained and the pH was stable in the range of 30-50°C at a pH of 5-7.

Trayectorias en pacientes con cáncer / Trajectories in cancer patients

INTRODUCCIÓN Este estudio busca generar conocimiento que informe políticas sanitarias que mejorar el acceso al diagnóstico de los pacientes con cáncer. OBJETIVO Describir la trayectoria de pacientes con cáncer de mama, pulmón y colon de 2 hospitales públicos de la Provincia de Buenos Aires desde el inicio de los síntomas hasta el diagnóstico de la enfermedad. PACIENTES Y MÉTODOS Se incluyeron personas con diagnóstico inicial de cáncer de mama, pulmón o colorrectal entre 1/3/2014 y el 29/6/2016 tratados en los hospitales Juan C Sanguinetti de Pilar y Alejandro Posadas de El Palomar. Los pacientes tuvieron cualquier estadio y estado funcional. Se excluyeron pacientes con recaídas o 2° tumores. Se realizó una revisión de historias clínicas, una encuesta telefónica para indagar tiempos, barreras y facilitadores y entrevistas en profundidad para caracterizar cualitativamente barreras y facilitadores. RESULTADOS Se revisaron 111 historias clínicas de pacientes del H. Sanguinetti. 14.4% se diagnosticaron por tamizaje y 85.6% como enfermedad sintomática. El 64.9% tuvo enfermedad localizada al diagnóstico y 35.1% enfermedad avanzada. Se realizaron 127 encuestas en ambos hospitales a 83 pacientes con cáncer de mama, 37 de colon y 7 de pulmón. 100 pacientes tuvieron enfermedad sintomática al diagnóstico y 27 fueron diagnosticados por tamizaje. El 28% consultó a más de 3 meses de iniciados los síntomas. Barreras referidas incluyen falta de jerarquización de los síntomas, miedo y problemas logísticos para turnos y consultas. Facilitadores referidos incluyen el conocimiento del hospital y de los profesionales y el apoyo de familiares y amigos. DISCUSIÓN En esta muestra, el diagnóstico de enfermedad sintomática y avanzada fue frecuente. Las barreras y facilitadores de la trayectoria son múltiples. Su reconocimiento brinda una oportunidad para establecer políticas activas para aumentar la conciencia sobre la enfermedad y allanar los obstáculos en al acceso al cuidado de la salud