The ESSENCE-Q: Can specialist supervision improve scoring agreement across specialist and public health nurses/nursery teachers?

AIM: This study aimed to explore whether the supervision of community public health nurses (PHNs) and nursery school teachers (NSTs) by a specialist, familiar with Early Symptomatic Syndromes Eliciting Neurodevelopmental Clinical Examinations (ESSENCE), improved the agreement of ESSENCE-Questionnaire (ESSENCE-Q) scoring, across raters. METHODS: A PHN, two NSTs, and a speech-language pathologist (SLP), familiar with ESSENCE, independently assessed 32 children. The ESSENCE-Q results were divided into the first (child 1-18) and second groups (the 19th child and the following children). Changes in score discrepancies were analysed for ESSENCE-Q cutoff scores and total ESSENCE-Q scores across raters. The SLP scores were used as a reference to evaluate sensitivity and specificity. RESULTS: The total ESSENCE-Q scores of the PHN and NSTs showed higher concordance in the second group (p < 0.05). Comparisons of the differences between the PHN/NSTs and SLP in total ESSENCE-Q scores showed a significantly smaller difference in the NSTs' scores in the second group (p < 0.05). CONCLUSION: The findings suggest that specialist supervision may lead to a better agreement between PHN and NSTs regarding ESSENCE-Q scores.

[Relationship between the number of public health nurses and the total number of people receiving home-visit nursing services for mental health/intractable diseases, and those numbers' regional differences].

Objectives　The primary aim of this study was to clarify the relationship between the number of public health nurses (PHNs) and the total number of people who received home-visit nursing services for mental health or intractable diseases. The secondary aim was to clarify the extent of regional differences in the number of PNHs and mental health or intractable diseases.Methods　This study used the total number of people who received home-visit nursing services for mental health or intractable diseases in 2019 from the Portal Site of Official Statistics of Japan (e-Stat) and population and area data in January 2020. Single and multiple regression analyses (covariates: population and area) were performed on the relationship between the number of PHNs per 100,000 population (abbreviated as "ratios of PHNs") and the total number of people who received home-visit nursing services for mental health or intractable diseases per 100,000 population (abbreviated as "mental health/intractable disease achievements"). Regional differences in ratios of PHNs and mental health/intractable disease achievement were examined using mean, standard deviation, maximum/minimum values, and Gini coefficients. Analyses were performed for each of the five units: the prefectures as a whole, prefectural public health centers, municipalities within the jurisdiction of prefectural public health centers, and cities in which public health centers are established (including or not including special wards).Results　Regression analyses indicated a positive relationship between the ratios of PHNs and mental health/intractable disease achievements. Multiple regression analysis indicated that both achievements were positively associated with population size and negatively associated with area size. The largest regression coefficients between the ratios of PHNs and achievements were 34.07 and 5.48 regarding mental health achievements and intractable disease achievements, respectively. For regional differences, the smallest Gini coefficient was the ratios of PHNs, and the largest was intractable disease achievements. The smallest and largest coefficient of the prefectures as a whole was 0.15 and 0.34, respectively. The maximum/minimum values of the prefectures as a whole also indicated that the smallest was 3.8 in the ratio of PHNs and the largest was 30.0 in intractable disease achievement.Conclusions　Increasing number of PHNs is needed to provide more home-visit nursing services for mental health and intractable diseases. It is particularly important to fill up the larger number of PHNs in smaller populations or larger area prefectures. Due to regional differences in the home-visit nursing service, it is important to promote the increase in the level of these activities.

Activin/Nodal/TGF-ß Pathway Inhibitor Accelerates BMP4-Induced Cochlear Gap Junction Formation During in vitro Differentiation of Embryonic Stem Cells.

Mutations in gap junction beta-2 (GJB2), the gene that encodes connexin 26 (CX26), are the most frequent cause of hereditary deafness worldwide. We recently developed an in vitro model of GJB2-related deafness (induced CX26 gap junction-forming cells; iCX26GJCs) from mouse induced pluripotent stem cells (iPSCs) by using Bone morphogenetic protein 4 (BMP4) signaling-based floating cultures (serum-free culture of embryoid body-like aggregates with quick aggregation cultures; hereafter, SFEBq cultures) and adherent cultures. However, to use these cells as a disease model platform for high-throughput drug screening or regenerative therapy, cell yields must be substantially increased. In addition to BMP4, other factors may also induce CX26 gap junction formation. In the SFEBq cultures, the combination of BMP4 and the Activin/Nodal/TGF-ß pathway inhibitor SB431542 (SB) resulted in greater production of isolatable CX26-expressing cell mass (CX26+ vesicles) and higher Gjb2 mRNA levels than BMP4 treatment alone, suggesting that SB may promote BMP4-mediated production of CX26+ vesicles in a dose-dependent manner, thereby increasing the yield of highly purified iCX26GJCs. This is the first study to demonstrate that SB accelerates BMP4-induced iCX26GJC differentiation during stem cell floating culture. By controlling the concentration of SB supplementation in combination with CX26+ vesicle purification, large-scale production of highly purified iCX26GJCs suitable for high-throughput drug screening or regenerative therapy for GJB2-related deafness may be possible.

Modeling gap junction beta 2 gene-related deafness with human iPSC.

There are >120 forms of non-syndromic deafness associated with identified genetic loci. In particular, mutation of the gap junction beta 2 gene (GJB2), which encodes connexin (CX)26 protein, is the most frequent cause of hereditary deafness worldwide. We previously described an induction method to develop functional CX26 gap junction-forming cells from mouse-induced pluripotent stem cells (iPSCs) and generated in vitro models for GJB2-related deafness. However, functional CX26 gap junction-forming cells derived from human iPSCs or embryonic stem cells (ESCs) have not yet been reported. In this study, we generated human iPSC-derived functional CX26 gap junction-forming cells (iCX26GJCs), which have the characteristics of cochlear supporting cells. These iCX26GJCs had gap junction plaque-like formations at cell-cell borders and co-expressed several markers that are expressed in cochlear supporting cells. Furthermore, we generated iCX26GJCs derived from iPSCs from two patients with the most common GJB2 mutation in Asia, and these cells reproduced the pathology of GJB2-related deafness. These in vitro models may be useful for establishing optimal therapies and drug screening for various mutations in GJB2-related deafness.

Generation of two iPSC lines from siblings of a homozygous patient with hearing loss and a heterozygous carrier with normal hearing carrying p.G45E/Y136X mutation in GJB2.

The gap junction beta-2 (GJB2) gene is the most common genetic cause of hereditary deafness worldwide. Among them, the G45E/Y136X mutation in GJB2 is the third most prevalent in Japan. In this study, we generated two induced pluripotent stem cell (iPSC) lines from peripheral blood mononuclear cells (PBMCs) of siblings with moderate-to-severe hearing loss (patient) or normal hearing (genetic carrier) carrying a homozygous or heterozygous G45E/Y136X mutation in GJB2 gene, respectively. These iPSC lines showed the expression of pluripotency markers and could differentiate into three germ layers. These disease-specific iPSC lines will be a powerful tool for investigating the pathogenesis of GJB2-related deafness.

Generation of two induced pluripotent stem cell lines from PBMCs of siblings carrying c.235delC mutation in the GJB2 gene associated with sensorineural hearing loss.

The gap junction beta-2 (GJB2) gene is the most common genetic cause of hereditary deafness worldwide. Especially, the 235delC mutation in GJB2 is most prevalent in East Asia. In this study, we generated two iPSC lines from PBMCs of siblings carrying homozygous 235delC mutation which exhibits an audiometric phenotype of profound hearing loss. These iPSC lines had normal karyotype, showed expression of pluripotency markers, and could differentiate into three germ layers. These disease specific iPSC lines may be useful for the construction of the disease models and for the elucidation of pathogenesis in GJB2-related deafness.

Generation of the induced pluripotent stem cell (hiPSC) line (JUFMDOi004-A) from a patient with hearing loss carrying GJB2 (p.V37I) mutation.

The gap junction beta-2 (GJB2) gene is the most common genetic cause of hereditary deafness worldwide. Especially, V37I mutation in GJB2 is most prevalent in Southeast Asia including Thailand, Malaysia, and Indonesia. Furthermore, it is the second most prevalent cause in Japan and China, and exhibits an audiometric phenotype of mild-to-moderate hearing loss. In this study, we generated induced pluripotent stem cells (iPSC) from peripheral blood mononuclear cells (PBMCs) of patient with homozygous V37I mutation. This iPSC line will be a powerful tool for investigating the pathogenesis and for developing a treatment for GJB2-related hearing loss.

Generation of Functional CX26-Gap-Junction-Plaque-Forming Cells with Spontaneous Ca2+ Transients via a Gap Junction Characteristic of Developing Cochlea.

Mutation of the gene GJB2, encoding connexin 26 (CX26; also known as gap junction beta 2), is the most frequent cause of hereditary deafness worldwide. CX26 is expressed in cochlear nonsensory cells, such as cochlear supporting cells, and forms gap junction plaques (GJPs) at cell-cell borders. Cochlear CX26-GJP-forming cells (Cx26GJCs) are thought to be an important therapeutic target for treatment of hereditary deafness. Nevertheless, the generation of Cx26GJCs-such as cochlear supporting cells-from embryonic stem/induced pluripotent stem (ES/iPS) cells has not been reported to date. Here, we detail a novel strategy for differentiating iPS cells into functional Cx26GJCs such as are found in cochlea. Several assays to characterize the phenotype of iPS-derived Cx26GJCs are described, including qRT-PCR, immunohistological analysis, morphological analysis, a scrape-loading and dye transfer assay, and calcium imaging. This in vitro model has applications in the establishment of inner-ear cell therapies and in drug screening to target GJB2-related hearing loss. © 2019 by John Wiley & Sons, Inc. Basic Protocol: Induction of mouse stem cells to create CX26-GJP-forming cells Support Protocol 1: Maintenance and passage of mouse induced pluripotent stem cells Support Protocol 2: Screening for high GJB2 and GJB6 expression in SFEBq culture using quantitative real-time PCR Support Protocol 3: Characterization of cells at different stages of differentiation by immunostaining Support Protocol 4: Ultrastructural analyses of cells at different stages of CX26-GJP-forming cell induction Support Protocol 5: Functional analyses of stem cell-derived CX26-GJP-forming cells.

In Vitro Models of GJB2-Related Hearing Loss Recapitulate Ca2+ Transients via a Gap Junction Characteristic of Developing Cochlea.

Mutation of the Gap Junction Beta 2 gene (GJB2) encoding connexin 26 (CX26) is the most frequent cause of hereditary deafness worldwide and accounts for up to 50% of non-syndromic sensorineural hearing loss cases in some populations. Therefore, cochlear CX26-gap junction plaque (GJP)-forming cells such as cochlear supporting cells are thought to be the most important therapeutic target for the treatment of hereditary deafness. The differentiation of pluripotent stem cells into cochlear CX26-GJP-forming cells has not been reported. Here, we detail the development of a novel strategy to differentiate induced pluripotent stem cells into functional CX26-GJP-forming cells that exhibit spontaneous ATP- and hemichannel-mediated Ca2+ transients typical of the developing cochlea. Furthermore, these cells from CX26-deficient mice recapitulated the drastic disruption of GJPs, the primary pathology of GJB2-related hearing loss. These in vitro models should be useful for establishing inner-ear cell therapies and drug screening that target GJB2-related hearing loss.

ESSENCE-Q - a first clinical validation study of a new screening questionnaire for young children with suspected neurodevelopmental problems in south Japan.

BACKGROUND: Early identification of autism spectrum disorder, intellectual developmental disorder, attention-deficit/hyperactivity disorder, and other neurodevelopmental disorders/problems is crucial, yet diagnosis is often delayed for years under the often misguided "wait-and-see" paradigm. The early symptomatic syndromes eliciting neurodevelopmental clinical examinations-questionnaire (ESSENCE-Q) is a brief (12-item) screening questionnaire developed specifically for the purpose of speeding up the identification process of a wide variety of neurodevelopmental problems. The aims were to 1) estimate the reliability of the ESSENCE-Q, 2) evaluate the clinical cutoff levels suggested by the author of the ESSENCE-Q, and 3) propose optimal cutoff levels based on receiver operating characteristic analysis. METHODS: The ESSENCE-Q was used for 1 year by a psychiatrist in Kochi, Japan, assessing children under the age of 6 years referred for developmental problems. The children were also clinically assessed with regard to whether or not they met criteria for a developmental disorder (diagnosis positive and diagnosis negative groups). We contrasted the results of the ESSENCE-Q and those of clinical diagnostic assessments in 130 cases. RESULTS: Cronbach's alpha was 0.82, sensitivity was 0.94 (95% confidence interval [CI]: [0.88, 0.98]), and specificity 0.53 (95% CI: [0.28, 0.77]), which are reasonable psychometrics for a first-step screening tool. Based on receiver operating characteristic analysis, we recommended an optimal cutoff level of yes ≥2 or maybe/a little ≥3 on the ESSENCE-Q (0.87 (95% CI: [0.79, 0.92]) sensitivity and 0.77 (95% CI: [0.50, 0.93]) specificity). CONCLUSION AND IMPLICATION: The ESSENCE-Q can be a good instrument for use as a screening tool for aiding in the process of early identification of neurodevelopmental disorders in clinical settings. To establish the broader validity and reliability of the ESSENCE-Q, case-control studies and general population studies of children in different age groups are needed.

Deformation of the Outer Hair Cells and the Accumulation of Caveolin-2 in Connexin 26 Deficient Mice.

BACKGROUND: Mutations in GJB2, which encodes connexin 26 (Cx26), a cochlear gap junction protein, represent a major cause of pre-lingual, non-syndromic deafness. The degeneration of the organ of Corti observed in Cx26 mutant-associated deafness is thought to be a secondary pathology of hearing loss. Here we focused on abnormal development of the organ of Corti followed by degeneration including outer hair cell (OHC) loss. METHODS: We investigated the crucial factors involved in late-onset degeneration and loss of OHC by ultrastructural observation, immunohistochemistry and protein analysis in our Cx26-deficient mice (Cx26f/fP0Cre). RESULTS: In ultrastructural observations of Cx26f/fP0Cre mice, OHCs changed shape irregularly, and several folds or notches were observed in the plasma membrane. Furthermore, the mutant OHCs had a flat surface compared with the characteristic wavy surface structure of OHCs of normal mice. Protein analysis revealed an increased protein level of caveolin-2 (CAV2) in Cx26f/fP0Cre mouse cochlea. In immunohistochemistry, a remarkable accumulation of CAV2 was observed in Cx26f/fP0Cre mice. In particular, this accumulation of CAV2 was mainly observed around OHCs, and furthermore this accumulation was observed around the shrunken site of OHCs with an abnormal hourglass-like shape. CONCLUSIONS: The deformation of OHCs and the accumulation of CAV2 in the organ of Corti may play a crucial role in the progression of, or secondary OHC loss in, GJB2-associated deafness. Investigation of these molecular pathways, including those involving CAV2, may contribute to the elucidation of a new pathogenic mechanism of GJB2-associated deafness and identify effective targets for new therapies.

[Municipal housing support for people with mental disabilities to facilitate transition from an institutional to community setting].

OBJECTIVES: The study investigated the provision of support by municipalities to assist people with mental disability securing a dwelling in the community. METHODS: In September 2008, a questionnaire was sent to welfare divisions concerned with people having mental disabilities in 1,805 municipalities nationwide. Responses were returned from 1,141 municipalities (recovery rate: 63.2%). The survey analyzed housing support, financial aid, understanding of transition and housing needs and recognition concerning health and welfare. RESULTS: Of those municipalities surveyed: 7.0% had a support system for those lacking a guarantor to sign a lease; 17.7% engaged in reducing the burden involved in securing a dwelling, mainly by setting up a housing advisory desk; and 5.0% worked with real-estate agents and housing providers to advance cooperation. Housing benefits were provided in 12.9% of municipalities for those living on welfare, with a special allowance equivalent to that for the physically challenged. Financial support such as funding to cover the rent and other relevant expenses was available in 2.5%. The number of people with mental disabilities seeking to make the transition to community living and the actual state of transition were not known in 52.9% of the municipalities; however, 64.9% considered that the amenities of a dwelling for such people should be comparable to those for the single-person household and the general public. Furthermore, 41.4% took actions to redress the negative images of people with mental disabilities held by the community residents. The assessment of policy regarding transition and housing support for people with mental disabilities revealed that 48.6% of municipalities considered a congregate living setting within the community, while acknowledgement of an independent living setting within the general neighborhood was low at 28.9%. Regarding the state of inter-ministerial collaboration between the welfare division for people with disabilities and other divisions, 25.5% indicated that support was provided in partnership with agencies beyond the immediate scope of health and welfare. CONCLUSION: A framework for the provision of municipality-managed transition and housing support for people with mental disability has yet to be fully developed on a national scale. A comprehensive policy regarding housing support is needed.

[Nationwide survey of municipality's maternal & child health promotion planning processes and consequent changes].

PURPOSE: The purpose of this study was to offer insight into the revisions needed for promotion of Maternal and Child Health Promotion Plans in the coming years. METHODS: The maternal and child health promotion planning process and changes in maternal and child health services were surveyed by questionnaires to all municipalities in Japan. RESULTS: Completed questionnaires were obtained from 2,362 municipalities (response rate: 72.6%) Replies from 2,202 municipalities, which had completed planning by March, 1998, were analyzed. Planning committees were established by 63.9% of municipalities, and working committees were set up in 55.4% of them. Members involved in the working committees were more limited when compared to the planning committees and only 9.3% of the committees had representatives from mothers. 37.5% of working committees held meetings more than 5 times. 56.0% of municipalities conducted hearings or questionnaire surveys of mothers and health personnel for assessment of maternal and child healthcare needs. Most prefectural public health centers provided statistical data for planning assistance. Public health center staff participated as planning members in 38.5% of municipalities, and a training program for the planning sponsored by health centers was utilized by 33.8% of municipalities. Only 18.3% of municipalities received support for explanation of the plans to the mayor and/or executive officers, only 12.1% received support for the management of the planning committees, and only 11.8% received support for needs analysis. Less than half of minicipalities conducted PR activities of the plans and progress management. A higher proportion conducted these activities in larger cities. After the planning process, 72.9% of municipalities started new projects according to the plans. However, 10.1% scrapped one or more projects. Changes in maternal and child healthcare services and promotion of coordination with other related organization were seen more in municipalities with a larger population. There were major differences found between prefectures with regards to the planning process, including the support from public health centers, and the effective use of plan reports, progress management, and changes in maternal and child healthcare services after the formulation of plans.

Investigation of indoor air pollution by chlorpyrifos: Determination of chlorpyrifos in indoor air and 3,5,6-trichloro-2-pyridinol in residents' urine as an exposure index.

OBJECTS: We carried out an investigation to clarify the real state of indoor air pollution by chlorpyrifos (termiticide) and exposure to chlorpyrifos of residents by measuring its urinary metabolite 3,5,6-trichloro-2-pyridinol (TCP) as an exposure index, such as biological monitoring. METHODS: The investigation was conducted in 43 individual houses with termiticide application (whether the termiticide was chlorpyrifos is uncertain) and 3 control houses without any termiticide application in Kagawa, Japan. Urine samples were collected from 46 healthy adult residents of the aforementioned houses. RESULTS: Chlorpyrifos in indoor air in the control houses was not detected (ND<1 ng/m(3), n=3), while 41 of 43 houses with termiticide application showed 1-350 ng/m(3). Although the chlorpyrifos concentrations in these 41 houses did not exceeded the indoor air quality guideline of 1000 ng/m(3), but 3 houses were higher than the guideline 100 ng/m(3) for children in Japan. Urinary TCP concentrations of 0.1-7.8 ng/mg·creatinine were detected in 41 residents from the 41 houses where chlorpyrifos had been detected. The chlorpyrifos concentration and the urinary TCP revealed a positive correlation (r=0.5468, p<0.01, n=41). CONCLUSIONS: The immediate health hazard from air born chlorpyrifos in the examined houses was negligible, but the findings suggest that it is necessary to monitor chemicals which may contaminate indoor air and to assess the risk of prolonged exposure to such chemicals. The measuring of urinary metabolite TCP of chlorpyrifos via biological monitoring would be useful, allowing comprehensive evaluation of the exposure to chlorpyrifos in indoor air.

[Levels of airborne dust by particle size in a suburb of Takamatsu, Japan].

PURPOSE: In recent years, attention has been paid to the influence of the particle fraction < or = 2.5 microns in diameter (PM2.5) among suspended particulate matter (SPM) on health. However, the present status of particle contamination in Japan has hitherto not been adequately clarified. To determine particle contamination in the outdoor air around Kagawa Medical School located on a hill in the suburb of Takamatsu City, we measured SPM according to diameter between February 1999 and January 2000. METHODS: Mass concentrations of SPM were determined using an Andersen air sampler and light scattering counts were measured using a particle counter at the same site. RESULTS: Of the entire year, the total SPM concentration (PM11) was 20-30 mg/m3, and the particle fraction < or = 2.1 microns in diameter (PM2.1) accounted for 25-60%. Both the PM11 and PM2.1 concentrations were the lowest in August when the wind blew mainly from rural areas, but high in January and February when the wind blew mainly from urban areas and April when yellow sand was wind borne from China. The SPM particle count in April was markedly high when a misty aerosol containing yellow sand was extent. The SPM count for each diameter class rapidly decreased after rainfall during passage of storm fronts, but this was especially marked for smaller diameters. During the measurement period in August, air from rural areas often passed the measurement point, and the particle concentration < or = 1 micron in diameter was markedly lower than that in April. CONCLUSION: The results suggest that the SPM levels around Kagawa Medical School located in the suburb of Takamatsu City are associated with seasonal climate and the wind direction from which the prevailing blew.