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CONTEXT: There are limited reports on the detailed examination of steroid profiles for setting algorithms for 21-hydroxylase deficiency (21OHD) screening by liquid chromatography-tandem mass spectrometry (LC-MS/MS). OBJECTIVE: We aimed to define an algorithm for newborn screening of 21OHD by LC-MS/MS, measuring a total of 2077 dried blood spot samples in Tokyo. METHODS: Five steroids (17α-hydroxyprogesterone [17αOHP], 21-deoxycortisol [21DOF], 11-deoxycortisol [11DOF], androstenedione [4AD], and cortisol [F]) were included in the panel of LC-MS/MS. Samples from 2 cohorts were assayed: Cohort A, 63 "screening positive" neonates who were referred to an endocrinologist (n = 26 with 21OHD; n = 37 false-positive; obtained from 2015 to 2020); and Cohort B, samples (n = 2014) with 17αOHP values in the 97th percentile or above, in the first-tier test with 17αOHP ELISA from 2020 to 2021. RESULTS: Analysis of Cohort A revealed that the 3 indexes 21DOF, 11DOF/17αOHP, and (4AD + 17αOHP)/F had higher area under the curve (AUC) values (0.999, 0.997, 0.989, respectively), while the 17αOHP AUC was lower (0.970). Accordingly, in addition to 17αOHP, the 3 markers were included for defining the screening algorithm. The assay of Cohort B revealed that the new algorithm gave 92% of predicted positive predictive value without false-negative cases. We also determined the reference values for the 5 steroids at 4 to 7 days after birth, according to sex and gestational age (GA), revealing extremely low levels of 21DOF at any GA irrespective of sex differences. CONCLUSION: Our study demonstrated the high relevance of 21DOF, (4AD + 17αOHP)/F, and 11DOF/17αOHP, rather than 17αOHP, for 21OHD screening.
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Hiperplasia Suprarrenal Congénita , Enfermedades del Sistema Endocrino , Recién Nacido , Femenino , Humanos , Masculino , Cortodoxona , Androstenodiona , Hidrocortisona , Cromatografía Liquida/métodos , Tamizaje Neonatal/métodos , Espectrometría de Masas en Tándem/métodos , 17-alfa-Hidroxiprogesterona , Hiperplasia Suprarrenal Congénita/diagnóstico , Esteroides , AndrógenosRESUMEN
PURPOSE: This study aims to examine the reliability and validity of Home-visit Nursing Quality Indicators for Children (HNQIC) with medical complexity in Japan that will enable measuring the quality of services provided by home-visit nursing agencies (HNA) for children with medical complexity (CMC) and their families. DESIGN AND METHODS: This study employed a model that measures medical quality as proposed by Donabedian in a conceptual framework. The HNQIC is comprised of a total of 42 items with responses in 5-point Likert scale: 8 items in "Structure", 24 items in "Process", and 10 items in "Outcome". A self-rating questionnaire survey was administered and responses from 57 home-visit nursing agencies were analyzed. An exploratory factor analysis was performed to examine the validity of the construct, and a covariance structure analysis was performed to examine the structural validity of the model that measures medical quality. RESULTS: The "Structure" and "Process" sections included 28 items in 5 factors, and the "Outcome" section included 7 items in 3 factors. The Cronbach's α coefficient for all of the items of "Structure" and "Process" was 0.93, and that of "Outcome" was 0.76. As a result of a covariance structure analysis, we obtained following goodness-of-fit indices: χ2 / df = 1.41, GFI = .897, AGFI = .794, CFI = .926, and for the coefficient of determination .14 ≤ R2 ≤ .68. CONCLUSIONS: As the statistical validity of the HNQIC was confirmed, we determined the goodness-of-fit indices of the model to be acceptable. PRACTICE IMPLICATIONS: The findings suggested that the HNQIC can be used as a quality indicator to access care effects objectively to provide better support.
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Indicadores de Calidad de la Atención de Salud , Niño , Análisis Factorial , Humanos , Japón , Psicometría , Reproducibilidad de los Resultados , Encuestas y CuestionariosRESUMEN
The number of rubella cases has increased in Japan, especially among adults. Rubella infection in pregnant females can lead to congenital rubella syndrome (CRS). The Japanese government is promoting vaccination to prevent CRS. This study performs a cost-effectiveness analysis of the following four methods: (1) females who wished to become pregnant and partners, with an antibody-titer test; (2) females only, with an antibody-titer test; (3) females and males, without an antibody-titer test; (4) females only, without an antibody-titer test. A decision tree model with a hypothetical cohort of 500 males and 500 females was used for the analysis, and the parameters were obtained from previous studies. The number of avoidances of CRS was defined as the effect. Compared to the case where methods were not implemented, the number of CRS cases that can be prevented by implementing the methods was 0.0115589 by (1) and (3) and 0.0147891 by (2) and (4). The cost effectiveness of (1) to (4) was 287,413,677 JPY, 135,050,529 JPY, 388,524,974 JPY, and 197,744,219 JPY, respectively (1 JPY = 0.00963247 USD). Method (2) was the most cost-effective and did not change by sensitivity analysis. We conclude that the vaccination for females only with an antibody-titer test is recommended.
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CONTEXT: We previously reported that inorganic iodine therapy in lactating women with Graves disease (GD) did not affect the thyroid function in 25 of 26 infants despite their exposure to excess iodine via breast milk. OBJECTIVE: To further assess thyroid function in infants nursed by mothers with GD treated with inorganic iodine. DESIGN: Case series. SETTING: Tajiri Thyroid Clinic, Japan. PARTICIPANTS: One hundred infants of lactating mothers with GD treated with potassium iodide (KI) for thyrotoxicosis. MAIN OUTCOME MEASURES: Infant blood thyrotropin (TSH) and free thyroxine (FT4) levels were measured by the filter paper method. Subclinical hypothyroidism was defined as TSH ≥10 µIU/mL and ≥5 µIU/mL in infants aged <6 and ≥6 months, respectively. RESULTS: Overall, 210 blood samples were obtained from 100 infants. The median infant age was 5 (range, 0-23) months; median maternal KI dose, 50 (4-100) mg/day; median blood TSH level, 2.7 (0.1-12.3) µIU/mL; and median blood FT4 level, 1.04 (0.58-1.94) ng/dL. Blood TSH level was normal in 88/100 infants. Twelve infants had subclinical hypothyroidism; among them, blood TSH levels normalized after maternal KI withdrawal or stopping breastfeeding in 3 infants. In 7 infants, blood TSH levels normalized during KI administration without stopping breastfeeding. Two infants could not be followed up. CONCLUSION: In Japan, inorganic iodine therapy for lactating women with GD did not affect thyroid function in most of the infants. Approximately 10% of infants had mild subclinical hypothyroidism, but blood TSH level normalized during continued or after discontinuing iodine exposure in all followed up infants.
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BACKGROUND: One of the major purposes of newborn screening for 21-hydroxylase deficiency (21OHD) is preventing life-threatening adrenal crisis. However, the details of adrenal crisis in newborns are not precisely documented. AIM: We aimed to clarify the clinical details of salt-wasting in newborn 21OHD patients. METHODS: Based on the follow-up survey of the screening in Tokyo from 1989 to 2017, we retrospectively analysed the conditions of classical 21OHD neonates before the initiation of therapy. RESULTS: One hundred classical 21OHD patients (55 male, 45 female) were analysed. The age at the first hospital visit was 0-20 days with sex difference (male: 9.0 ± 3.5 days; female: 6.2 ± 3.9 days). Thirty-seven (37.4%) patients exhibited severe salt-wasting (SSW), that is, Na < 130 mEq/L, K > 7 mEq/L or Na/K ratio < 20; except for one case, SSW developed in or after the second week of life. The serum concentrations of Na, K and Na/K were linearly correlated with age in days (R2 = .38, .25, and .34 respectively), suggesting that the risk of SSW increases linearly without a threshold. The age at which the regression lines reached Na < 130 mEq/L, K > 7 mEq/L and Na/K < 20 was approximately coincided, 11.1, 12.3 and 11.2 days, respectively. All SSW patients exhibited decreased body weight from birth in their second week of life. CONCLUSION: Our data revealed that the risk of developing SSW increases during the second week of life without a threshold, and for preventing SSW, early intervention, ideally during first week of life, is desirable. An increased body weight in the second week of life indicates the absence of SSW.
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Hiperplasia Suprarrenal Congénita , Peso Corporal , Femenino , Humanos , Recién Nacido , Masculino , Tamizaje Neonatal , Estudios Retrospectivos , Esteroide 21-HidroxilasaRESUMEN
The Nankai Trough in Japan has been identified as a geological feature that could cause extensive damage in the event of a major earthquake. In this study, we investigated the impact of effective guidance signs for hearing-impaired individuals requiring special care when escaping to a tsunami evacuation building (emergency evacuation location) using geographical information system (GIS) and viewshed analysis. We selected an area we considered would suffer severe damage following a major earthquake and tsunami; we identified difficulties in the provision of escape routes. Using GIS, we determined the time required for escape and tsunami arrival time if effective signs were installed; we undertook such analysis using the height data of buildings in the target area. With effectively installed guidance signs, the average required evacuation time was 36.88 minutes; without such signs (which is currently the case in the target area), the average time was 47.10 minutes: that would result in citizens getting caught by the tsunami. Installing effective guidance signs would allow hearing-impaired individuals to escape to an evacuation building before being hit by the tsunami.
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Planificación en Desastres/métodos , Sistemas de Información Geográfica , Directorios de Señalización y Ubicación , Personas con Deficiencia Auditiva , Tsunamis , Planificación en Desastres/normas , Humanos , JapónRESUMEN
CONTEXT: The effects of maternal inorganic iodine therapy on infant thyroid function are not well known. OBJECTIVE: This study investigated the effects on infant thyroid function of maternal inorganic iodine therapy when administered to lactating mothers with Graves disease. DESIGN AND SETTING: This study was a prospective case series performed at the Tajiri Thyroid Clinic, Kumamoto, Japan. PARTICIPANTS: Subjects were 26 infants of lactating mothers with Graves disease treated with potassium iodide (KI) for postpartum thyrotoxicosis. MAIN OUTCOME MEASURES: Infant blood levels of thyroid-stimulating hormone (TSH) and free thyroxine were measured using the dried filter-paper method. Iodine concentrations in breast milk and infant urine were measured on the same day. Subclinical hypothyroidism was defined as a blood TSH level of ≥10 or ≥5 µIU/mL in <6-month-old and 6- to 12-month-old infants, respectively. RESULTS: The median age of the infants was 3 months (range, 0 to 10 months). The median KI dose was 50 mg/d (range, 10 to 100 mg/d). High median iodine concentrations were detected in breast milk (15,050 µg/L; range, 831 to 72,000 µg/L) and infant urine (15,650 µg/L; range, 157 to 250,000 µg/L). Twenty-five of 26 infants had normal thyroid function. Although one infant had subclinical hypothyroidism (blood TSH, 12.3 µIU/mL), the TSH level normalized to 2.3 µIU/mL at 2 months after KI discontinuation. CONCLUSION: In Japan, where iodine intake is sufficient, administration of inorganic iodine to lactating mothers with Graves disease did not affect thyroid function in most infants despite high levels of exposure to iodine via breast milk.
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BACKGROUND: Congenital adrenal hyperplasia (CAH) cause life-threatening adrenal crisis. It also affects fetal sex development and can result in incorrect sex assignment at birth. In 1989, a newborn screening program for congenital adrenal hyperplasia (CAH) was introduced in Tokyo. Here we present the results of this screening program in order to clarify the efficiency of CAH screening and the incidence of CAH in Japan. METHOD: From 1989 to 2013, a total of 2,105,108 infants were screened for CAH. The cutoff level for diagnosis of CAH was adjusted for gestational age and birth weight. RESULTS: A total of 410 infants were judged positive, and of these, 106 patients were diagnosed with CAH, indicating a positive predictive value (PPV) of 25.8 %. Of the 106 patients, 94 (88.7 %) were diagnosed with 21-OHD. Of these 94 patients, 73 were diagnosed with the salt wasting form, 14 with the simple virilising form and 7 with the nonclassical form (NC21OHD). The mean birth weight and gestational age were 3192 ± 385 g and 38.9 ± 1.38 weeks. 11 out of 44 female patients were assigned as female according to their screening result. CONCLUSIONS: These data suggest that the newborn screening in Tokyo was effective, especially for sex assignment and preventing fatal adrenal crisis. The incidence of CAH was similar to that measured in previous Japanese screening studies, and it was also similar to that of western countries. The incidence of NC21OHD in Japan in the present study was lower than that in western countries as previous studies reported. The screening program achieved higher PPV than previous CAH screening studies, which might be due to the use of variable cutoffs according to gestational age and birth weight. However, most of the neonates born at 37 weeks or less that were referred to hospital were false-positives. Further changes are needed to reduce the number of false positive preterm neonates.
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Hiperplasia Suprarrenal Congénita/epidemiología , Tamizaje Neonatal , 17-alfa-Hidroxiprogesterona/sangre , Hiperplasia Suprarrenal Congénita/diagnóstico , Peso al Nacer , Edad Gestacional , Humanos , Incidencia , Recién Nacido , Tamizaje Neonatal/métodos , Valor Predictivo de las Pruebas , Estudios Retrospectivos , Tokio/epidemiologíaRESUMEN
To develop a covalent and specific labeling method for single- and double-stranded plasmid DNA, photoresponsive oligonucleotide containing 3-cyanovinylcarbazole nucleoside was adopted. Single- and double-stranded plasmid DNA was successfully labeled/de-labeled with Cy3 and/or biotin by photoirradiation.
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ADN/química , Oligonucleótidos/química , Plásmidos/genética , Coloración y Etiquetado/métodos , Acrilonitrilo/análogos & derivados , Acrilonitrilo/química , Carbazoles/química , Colorantes Fluorescentes/química , Procesos FotoquímicosRESUMEN
DNA-loaded microtubules (MTs) moving on a kinesin motor protein-coated substrate can selectively hybridize with a target fully matched DNA over single-base mismatched DNA and transport it. This technique is capable of collecting target biomolecules toward one point site to design new methodology of DNA analysis.
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Proteínas Bacterianas/química , Biotina/análogos & derivados , Sondas de ADN , ADN/aislamiento & purificación , Cinesinas/química , Proteínas Asociadas a Microtúbulos/química , Adsorción , Animales , Biotina/química , Microscopía Fluorescente , PorcinosRESUMEN
Kinesin is a linear motor protein driven by energy released by ATP hydrolysis. In the present work, we genetically installed an M13 peptide sequence into Loop 12 of kinesin, which is one of the major microtubule binding regions of the protein. Because the M13 sequence has high affinity for Ca(2+)-calmodulin, the association of the engineered kinesin with microtubules showed a steep Ca(2+)-dependency in ATPase activity at Ca(2+) concentrations of pCa 6.5-8. The calmodulin-binding domain of plant kinesin-like calmodulin-binding protein is also known to confer Ca(2+)-calmodulin regulation to kinesins. Unlike this plant kinesin, however, our novel engineered kinesin achieves this regulation while maintaining the interaction between kinesin and microtubules. The engineered kinesin is switched on/off reversibly by an external signal (i.e., Ca(2+)-calmodulin) and, thus, can be used as a model system for a bio/nano-actuator.
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Calcio/farmacología , Cinesinas/genética , Cinesinas/metabolismo , Ingeniería de Proteínas , Secuencia de Aminoácidos , Calcio/química , Calmodulina/metabolismo , Cationes Bivalentes/química , Cationes Bivalentes/farmacología , Humanos , Cinesinas/química , Microtúbulos/metabolismo , Modelos Moleculares , Datos de Secuencia Molecular , Estructura Terciaria de ProteínaRESUMEN
We measured saccadic response time (SRT) to investigate developmental changes related to spatially aligned or misaligned auditory and visual stimuli responses. We exposed 4-, 5-, and 11-month-old infants to ipsilateral or contralateral auditory-visual stimuli and monitored their eye movements using an electro-oculographic (EOG) system. The SRT analyses revealed four main results. First, saccades were triggered by visual stimuli but not always triggered by auditory stimuli. Second, SRTs became shorter as the children grew older. Third, SRTs for the ipsilateral and visual-only conditions were the same in all infants. Fourth, SRTs for the contralateral condition were longer than for the ipsilateral and visual-only conditions in 11-month-old infants but were the same for all three conditions in 4- and 5-month-old infants. These findings suggest that infants acquire the function of auditory-visual spatial integration underlying saccadic eye movement between the ages of 5 and 11 months. The dependency of SRTs on the spatial configuration of auditory and visual stimuli can be explained by cortical control of the superior colliculus. Our finding of no differences in SRTs between the ipsilateral and visual-only conditions suggests that there are multiple pathways for controlling the superior colliculus and that these pathways have different developmental time courses.
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Estimulación Acústica , Percepción Auditiva/fisiología , Desarrollo Infantil/fisiología , Tiempo de Reacción , Movimientos Sacádicos/fisiología , Percepción Espacial/fisiología , Percepción Visual/fisiología , Electrooculografía , Femenino , Humanos , Lactante , Masculino , Monitoreo Fisiológico , Estimulación Luminosa , Colículos Superiores/fisiología , TiempoRESUMEN
We have developed a novel mobile bioprobe using a conjugate of a kinesin-driven microtubule (MT) and malachite green (MG) as a platform for capturing MG RNA aptamers. The fluorescence of MG increases when it is bound to an MG aptamer, allowing MT-MG conjugates to work as sensors of RNA transcripts containing the MG aptamer sequence. Kinesin motor proteins provide an effective driving force to create mobile bioprobes without any manipulation. Although the fluorescence of a small number of MG-binding aptamers is low, the self-organization of tubulins into MTs enables the microscopic observation of the bound aptamers by collecting them on MTs. We demonstrate that MT-MG conjugates can select target aptamers from a transcription mixture and transport them without losing their inherent motility. Because the MG aptamer binds MG in a reversible manner, MT-MG conjugates can conditionally load and unload the target aptamers. This is one advantage of this system over the molecular probes developed previously in which reversible unloading is impossible due to high-affinity binding, such as between avidin and biotin. Furthermore, an MT-MG conjugate can be used as a platform for other MG aptameric sensors with recognition regions for various target analytes optimized by further selection procedures. This is the first step to applying living systems to in vitro devices. This technique could provide a new paradigm of mobile bioprobes establishing high-throughput in vitro selection systems using microfluidic devices operating in parallel.
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Aptámeros de Nucleótidos/química , Cinesinas/química , Sondas ARN/química , Colorantes de Rosanilina/química , Técnicas Analíticas MicrofluídicasRESUMEN
To examine the functional role of the essential light chain (ELC) in the phosphorylation-dependent regulation of smooth muscle myosin, we replace the native light chain in smooth muscle myosin with bacterially expressed chimeric ELCs in which one or two of the four helix-loop-helix domains of chicken gizzard ELC were substituted by the corresponding domains of scallop (Aquipecten irradians) ELC. All of these myosins, regardless of the ELC mutations or regulatory light chain (RLC) phosphorylation, showed normal subunit constitutions and NH(4)(+)/EDTA-ATPase activities, both of which were similar to those of native myosin. None of the ELC mutations changed the actin-activated ATPase activity of myosin in the absence of RLC phosphorylation. However, in the presence of RLC phosphorylation, the substitution of domain 1 or 2 in the ELC significantly decreased the actin-activated ATPase activity, whereas the substitution of both of these domains did not change the activity. In contrast to myosin, the domain 2 substitution in the ELC did not affect the actin-activated ATPase activity of single-headed myosin subfragment 1. These results suggest an interhead interaction between domains 1 and 2 of ELCs which is required to attain the full actin-activated ATPase activity of smooth muscle myosin in the presence of RLC phosphorylation.