RESUMEN
Spinal muscular atrophy was recently added to the Wisconsin newborn screening panel. Here we report our screening methods, algorithm, and outcomes. A multiplex real-time PCR assay was used to identify newborns with homozygous SMN1 exon 7 deletion, and those newborns' specimens further underwent a droplet digital PCR assay for SMN2 copy number assessment. An independent dried blood spot specimen was collected and tested to confirm the initial screening results for SMN1 and SMN2. From October 15, 2019 to October 14, 2020, a total of 60,984 newborns were screened for spinal muscular atrophy. Six newborns screened positive for and were confirmed to have spinal muscular atrophy, making the Wisconsin spinal muscular atrophy birth prevalence 1 in 10,164. Of these six infants, two have two copies of SMN2, two have three copies of SMN2, and two have four copies of SMN2. Five newborns received Zolgensma therapy, and one newborn received Spinraza therapy. Our screening method's positive predictive value is 100%. This comprehensive approach, providing both timely SMN2 information and SMN1 and SMN2 confirmation as parts of the algorithm for spinal muscular atrophy newborn screening, facilitated timely clinical follow-up, family counseling, and treatment planning.
Asunto(s)
Atrofia Muscular Espinal , Atrofias Musculares Espinales de la Infancia , Homocigoto , Humanos , Lactante , Recién Nacido , Atrofia Muscular Espinal/diagnóstico , Atrofia Muscular Espinal/genética , Tamizaje Neonatal , Atrofias Musculares Espinales de la Infancia/diagnóstico , Atrofias Musculares Espinales de la Infancia/genética , Proteína 1 para la Supervivencia de la Neurona Motora/genética , Wisconsin/epidemiologíaRESUMEN
All newborn screening programs screen for severe combined immunodeficiency by measurement of T-cell receptor excision circles (TRECs). Herein, we report our experience of reporting TREC assay results as multiple of the median (MoM) rather than using conventional copy numbers. This modification simplifies the assay by eliminating the need for standards with known TREC copy numbers. Furthermore, since MoM is a measure of how far an individual test result deviates from the median, it allows normalization of TREC assay data from different laboratories, so that individual test results can be compared regardless of the particular method, assay, or reagents used.
RESUMEN
BACKGROUND: Identifying congenital hypothyroidism through newborn screening (NBS) is higher among moderate-to-late preterm (MLPT) infants. Currently, the same thyroid-stimulating hormone (TSH) cutoffs are used for term and preterm infants. TSH reference ranges for MLPT infants are not currently available. OBJECTIVE: To determine TSH reference ranges for MLPT infants. METHODS: We analyzed 10,987 TSH levels on NBS samples performed on 8499 MLPT infants born between 32 and 36 weeks gestation. RESULTS: TSH median, 5th, 25th, 75th, 95th, and 99th percentiles were defined from day 1 until day 14 of life. TSH levels gradually decreased after birth and reached a plateau around day 6. CONCLUSION: Using a state-wide cohort, we constructed TSH reference charts from day 1 until day 14 for MLPT infants. Relationship between age-adjusted TSH percentiles and long-term neurodevelopmental outcomes should be determined in future studies to define optimal TSH cutoffs for MLPT infants.
Asunto(s)
Hipotiroidismo Congénito , Tirotropina , Hipotiroidismo Congénito/diagnóstico , Hipotiroidismo Congénito/epidemiología , Edad Gestacional , Humanos , Lactante , Recién Nacido , Recien Nacido Prematuro , Valores de ReferenciaRESUMEN
BACKGROUND AND OBJECTIVES: Many newborn screening (NBS) programs now perform repeat or serial NBS to detect congenital hypothyroidism. There is wide variation in thyroid-stimulating hormone (TSH) cutoffs used by NBS programs. Data on TSH reference ranges in preterm infants at increasing postnatal age are limited. Our study objective was to determine TSH reference ranges for preterm infants born at <32 weeks' gestation. METHODS: We analyzed serial TSH levels on NBS performed on infants born between 22 and 31 weeks' gestation from 2012 to 2016 in Wisconsin. The study cohort was divided into 2 groups (22-27 and 28-31 weeks), and TSH percentiles were defined from birth to the term equivalent gestational age. RESULTS: The study cohort consisted of 1022 and 2115 infants born at 22 to 27 and 28 to 31 weeks' gestation, respectively. The 95th percentile TSH level for the group born at 22 to 27 weeks' gestation gradually decreased and reached a nadir at â¼10 to 11 weeks. In contrast, for the group born at 28 to 31 weeks' gestation, the 95th percentile TSH level reached a nadir at â¼5 to 6 weeks. At 3 to 4 weeks after birth, the 95th percentile TSH level ranged from 11 to 11.8 µIU/mL for the group born at 22 to 27 weeks' gestation and ranged from 8.2 to 9 µIU/mL for the group born at 28 to 31 weeks' gestation. CONCLUSIONS: Using a statewide cohort of preterm infants, we constructed TSH reference charts from birth to the term equivalent gestation for preterm infants born at <32 weeks' gestation. Use of a single cutoff for all preterm infants might lead to misdiagnosis. The differences in TSH levels according to gestational-age categories might explain the increased frequency in congenital hypothyroidism diagnoses among preterm infants. These data are useful for defining age-adjusted NBS TSH cutoffs for preterm infants.
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Recien Nacido Prematuro/sangre , Tirotropina/sangre , Biomarcadores/sangre , Estudios de Cohortes , Femenino , Humanos , Recién Nacido , Masculino , Valores de ReferenciaRESUMEN
OBJECTIVES: To determine the incidence of congenital hypothyroidism in preterm infants and to identify associated risk factors. STUDY DESIGN: A population-based cohort study was performed in preterm infants born at <32 weeks of gestational age between 2012 and 2016 in Wisconsin. Newborn screening (NBS) results and demographic data were obtained from the Wisconsin State Laboratory of Hygiene. Congenital hypothyroidism was subdivided to early TSH elevation (eTSH) and delayed TSH elevation (dTSH). Multivariate logistic regression analyses were performed to identify demographic factors associated with dTSH. RESULTS: A total of 3137 preterm infants born at 22-31 weeks of gestational age were included in the study. Mean gestational age was 28.4 ± 2.4 weeks and mean birth weight was 1191 ± 399 g. Forty-nine infants were diagnosed with congenital hypothyroidism. The overall incidence of congenital hypothyroidism was 1.56%, including a 0.13% incidence of eTSH and a 1.43% incidence of dTSH. Birth weight <1000 g, multiple gestation, and initial TSH level were identified as independent predictors for dTSH. CONCLUSION: Targeted serial NBS in Wisconsin led to a higher rate of diagnosis of congenital hypothyroidism in preterm infants than has been reported previously. The majority (92%) of congenital hypothyroidism cases were diagnosed with dTSH. Birth weight <1000 g, multiple gestation, and elevated initial TSH level were associated with increased risk for development of dTSH. We recommend obtaining targeted serial NBS in preterm infants (<32 weeks of gestational age) to improve the detection of congenital hypothyroidism.
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Hipotiroidismo Congénito/diagnóstico , Tamizaje Neonatal/métodos , Tirotropina/sangre , Biomarcadores/sangre , Hipotiroidismo Congénito/sangre , Hipotiroidismo Congénito/epidemiología , Femenino , Estudios de Seguimiento , Edad Gestacional , Humanos , Incidencia , Recién Nacido , Recien Nacido Prematuro , Masculino , Vigilancia de la Población/métodos , Estudios Retrospectivos , Factores de Riesgo , Wisconsin/epidemiologíaRESUMEN
The p15 fusion-associated small transmembrane (FAST) protein is a nonstructural viral protein that induces cell-cell fusion and syncytium formation. The exceptionally small, myristoylated N-terminal ectodomain of p15 lacks any of the defining features of a typical viral fusion protein. NMR and CD spectroscopy indicate this small fusion module comprises a left-handed polyproline type II (PPII) helix flanked by small, unstructured N and C termini. Individual prolines in the 6-residue proline-rich motif are highly tolerant of alanine substitutions, but multiple substitutions that disrupt the PPII helix eliminate cell-cell fusion activity. A synthetic p15 ectodomain peptide induces lipid mixing between liposomes, but with unusual kinetics that involve a long lag phase before the onset of rapid lipid mixing, and the length of the lag phase correlates with the kinetics of peptide-induced liposome aggregation. Lipid mixing, liposome aggregation, and stable peptide-membrane interactions are all dependent on both the N-terminal myristate and the presence of the PPII helix. We present a model for the mechanism of action of this novel viral fusion peptide, whereby the N-terminal myristate mediates initial, reversible peptide-membrane binding that is stabilized by subsequent amino acid-membrane interactions. These interactions induce a biphasic membrane fusion reaction, with peptide-induced liposome aggregation representing a distinct, rate-limiting event that precedes membrane merger. Although the prolines in the proline-rich motif do not directly interact with membranes, the PPII helix may function to force solvent exposure of hydrophobic amino acid side chains in the regions flanking the helix to promote membrane binding, apposition, and fusion.
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Lipoilación , Modelos Químicos , Ácido Mirístico/química , Péptidos/química , Reoviridae/química , Proteínas Virales de Fusión/química , Secuencias de Aminoácidos , Animales , Chlorocebus aethiops , Liposomas/química , Liposomas/metabolismo , Ácido Mirístico/metabolismo , Resonancia Magnética Nuclear Biomolecular , Péptidos/síntesis química , Péptidos/genética , Péptidos/metabolismo , Estructura Terciaria de Proteína , Reoviridae/genética , Reoviridae/metabolismo , Células Vero , Proteínas Virales de Fusión/genética , Proteínas Virales de Fusión/metabolismoRESUMEN
The S1 genome segments of avian and Nelson Bay reovirus encode tricistronic mRNAs containing three sequential partially overlapping open reading frames (ORFs). The translation start site of the 3'-proximal ORF encoding the sigmaC protein lies downstream of two ORFs encoding the unrelated p10 and p17 proteins and more than 600 nucleotides distal from the 5'-end of the mRNA. It is unclear how translation of this remarkable tricistronic mRNA is regulated. We now show that the p10 and p17 ORFs are coordinately expressed by leaky scanning. Translation initiation events at these 5'-proximal ORFs, however, have little to no effect on translation of the 3'-proximal sigmaC ORF. Northern blotting, insertion of upstream stop codons or optimized translation start sites, 5'-truncation analysis, and poliovirus 2A protease-mediated cleavage of eIF4G indicated sigmaC translation derives from a full-length tricistronic mRNA using a mechanism that is eIF4G-dependent but leaky scanning- and translation reinitiation-independent. Further analysis of artificial bicistronic mRNAs failed to provide any evidence that sigmaC translation derives from an internal ribosome entry site. Additional features of the S1 mRNA and the mechanism of sigmaC translation also differ from current models of ribosomal shunting. Translation of the tricistronic reovirus S1 mRNA, therefore, is dependent both on leaky scanning and on a novel scanning-independent mechanism that allows translation initiation complexes to efficiently bypass two functional upstream ORFs.
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Codón Iniciador/metabolismo , Genes/fisiología , Orthoreovirus Aviar/metabolismo , Iniciación de la Cadena Peptídica Traduccional/fisiología , ARN Viral/metabolismo , Ribosomas/metabolismo , Proteínas Virales/biosíntesis , Animales , Línea Celular , Codón Iniciador/genética , Factor 4G Eucariótico de Iniciación/genética , Factor 4G Eucariótico de Iniciación/metabolismo , Genoma Viral , Modelos Biológicos , Sistemas de Lectura Abierta/fisiología , Orthoreovirus Aviar/genética , Poliovirus/genética , Poliovirus/metabolismo , Codorniz , ARN Viral/genéticaRESUMEN
Enterovirus 70 (EV70) is one of several human enteroviruses that exhibit a propensity for infecting the central nervous system (CNS). The mechanisms by which neurotropic enteroviruses gain access to and invade the CNS are poorly understood. One possibility is that circulating leukocytes become infected and carry neurotropic enteroviruses to the CNS. We examined the ability of EV70 to infect cell lines derived from lymphoid, myeloid, and monocytic lineages. Most leukocyte cell lines tested bound radiolabeled EV70 and were permissive for EV70 replication, suggesting that EV70, in contrast to other enteroviruses, has an in vitro tropism that includes lymphoid, monocytic, and myeloid cell lines. For some of the cell lines, virus binding and infection correlated with surface expression of decay-accelerating factor (DAF), an attachment protein for EV70 on HeLa cells. However, EV70 also adsorbed to and infected cell lines that expressed little or no DAF. In contrast to what was observed for HeLa cells, neither DAF-specific monoclonal antibodies nor phosphatidylinositol-specific phospholipase C treatment inhibited EV70 binding to permissive leukocyte cell lines, and antibody blockade of DAF had little or no effect on EV70 replication. We also found that neither the human coxsackievirus-adenovirus receptor nor intercellular cell adhesion molecule 1, which mediate the entry of coxsackie B viruses and coxsackievirus A21, respectively, functions as a receptor for EV70. EV70 binding to all cell lines was sensitive to sialidase treatment and to inhibition of O glycosylation by benzyl N-acetyl-alpha-D-galactosaminide. Taken together, these results suggest that a sialylated molecule(s) other than DAF serves as a receptor for EV70 on permissive human leukocyte cell lines.