Whole‐exome sequencing of a unique brain malformation with periventricular heterotopia,cingulate polymicrogyria and midbrain tectal hyperplasia

We report a case of an infant with unique and unreported combinations of brain anomalies. The patient showed distinctive facial findings, severe delay in psychomotor development, cranial nerve palsy and seizures. Brain magnetic resonance imaging performed at 5 days of age revealed complex brain malformations, including heterotopia around the mesial wall of lateral ventricles, dysmorphic cingulate gyrus, and enlarged midbrain tectum. The patient unexpectedly died at 13 months of age. Postmortem pathological findings included a polymicrogyric cingulate cortex, periventricular nodular heterotopia, basal ganglia and thalamic anomalies, and dysmorphic midbrain tectum. Potential candidate genes showed no abnormalities by traditional PCR‐based sequencing. Whole‐exome sequencing confirmed the presence of novel gene variants for filamin B (FLNB), guanylate binding protein family member 6, and chromosome X open reading frame 59, which adapt to the autosomal recessive mode or X‐linked recessive mode. Although immunohistochemical analysis confirmed the expression of FLNB protein in the vessel walls and white matter in autopsied specimens, there may be functional relevance of the compound heterozygous FLNB variants during brain development.     

Development of photo-amino acid derivative reactivity assay: a novel in chemico alternative method for predicting photoallergy

Photoallergy test of cosmetics and several types of pharmaceutical substances is often necessary for obtaining approval from authorities. However, there are no official test guidelines for photoallergy evaluation. Therefore, we tried to establish a photoallergy test by utilizing an in chemico alternative sensitization method, amino acid derivative reactivity assay (ADRA). To determine the criteria for judging the photoallergy potential, photo-ADRA with or without photoirradiation was performed using 60 photoallergenic chemicals, and cysteine and lysine derivatives were detected using high-performance liquid chromatography either by absorbance or fluorescence measurement. The accuracy of prediction was 81.4% (48 of 59) and 80.0% (48 of 60) using the absorbance and fluorescence methods, respectively. However, as chemicals can breakdown into multiple chemicals during photoirradiation, the absorbance method often cannot perform accurate detection due to co-elution, whereas the fluorescence method can do this due to lack of co-elution. Moreover, all eight chemicals that were found to be negative or false-positive for photoirritation in the 3T3 neutral red uptake phototoxicity test were confirmed as positive for photoallergy using this method. Furthermore, we prepared three types of pseudo-mixtures where we added one photoallergen along with five nonphotoallergens and performed the photo-ADRA by the ultraviolet and fluorescence methods. The result of the fluorescence method was almost the same as that obtained with the use of a single photoallergen and hence the outcome was not affected by the mixture. Thus, this study not only showed a method of evaluating the photoallergy potential of a single chemical but also a mixture, making it useful as an in chemico photoallergy alternative test.     

Intracellular translocation of glutathione S-transferase pi during oligodendrocyte differentiation in adult rat cerebral cortex in vivo

Glutathione S-transferase (GST)-pi is a cytosolic isoenzyme used as a marker for mature oligodendrocytes in the mammalian brain. However, the cellular properties of GST-pi-immunoreactive [GST-pi (+)] cells in adult brain are not completely understood. We immunohistochemically demonstrated the existence of two subtypes of GST-pi (+) cells in the cerebral cortex of adult rats: one subtype exhibited GST-pi in the cytoplasm (C-type cells), while the other did mainly in the nucleus (N-type cells). The GST-pi (+) C-type cells were also immunopositive for 2',3'-cyclic nucleotide 3'-phosphodiesterase and RIP, indicating that they were mature oligodendrocytes, while the GST-pi (+) N-type cells expressed NG2, indicating that they were oligodendrocyte progenitor cells. Furthermore, observation of the fate of newly-generated cells by 5-bromodeoxyuridine-labeling revealed that the GST-pi (+) N-type cells differentiated into C-type cells. These findings indicate translocation of GST-pi from the nucleus to the cytoplasm during oligodendrocyte maturation.     

Dynamic changes in conjunctival dendritic cell numbers, anatomical position and phenotype during experimental allergic conjunctivitis

Dendritic cells (DCs) are a subset of antigen-presenting cells (APCs) that are involved in the initiation and control of the immune response to antigens present at the interface with the environment. A limited number of groups have studied DCs in human and animal conjunctiva but no data is available concerning the different DC subsets present in the conjunctival tissue. The aims of this study are to characterize the phenotypes and numbers of DCs present in the murine model of allergic conjunctivitis using the technique of immunohistochemistry so as to aid the understanding of the mechanisms involved in allergic eye disease. A double immunofluorescence method was used to analyze the phenotypic distribution and density of DC subsets in the mouse conjunctival tissues of the allergic model using a panel of antibodies: CD11c, as a general marker of DCs, coupled with another DC subset marker such as Langerin for Langerhans cells (LCs), CD11b for myeloid DCs (mDCs) and mPDCA-1 for plasmacytoid DCs (pDCs). In the na?ve conjunctiva, mDCs were consistently detected in the subepithelial layer and substantia propria. In the epithelium and the subepithelial layer, very few LCs and virtually no pDCs were observed. Following allergen challenge, there was a marked influx of mDCs and pDCs, but no LCs, into the subepithelial layer and throughout the substantia propria. These results indicate that conjunctival DC subsets may play an important role in the immune-regulatory processes involved in the inflammatory component of allergic conjunctivitis.     

Lifestyle factors and social ties associated with the frequency of laughter after the Great East Japan Earthquake: Fukushima Health Management Survey

Purpose

Although mental health problems such as depression after disasters have been reported, positive psychological factors after disasters have not been examined. Recently, the importance of positive affect to our health has been recognised. We therefore investigated the frequency of laughter and its related factors among residents of evacuation zones after the Great East Japan Earthquake of 2011.

Methods

In a cross-sectional study on 52,320 participants aged 20 years and older who were included in the Fukushima Health Management Survey in Japan’s fiscal year 2012, associations of the frequency of laughter with changes in lifestyle after the disaster, such as a changed work situation, the number of family members, and the number of address changes, and other sociodemographic, psychological, and lifestyle factors were examined using logistic regression analysis. The frequency of laughter was assessed using a single-item question: “How often do you laugh out loud?”

Results

The proportion of those who laugh almost every day was 27.1%. Multivariable models adjusted for sociodemographic, psychological, and lifestyle factors demonstrated that an increase in the number of family members and fewer changes of address were significantly associated with a high frequency of laughter. Mental health, regular exercise, and participation in recreational activities were also associated with a high frequency of laughter.

Conclusion

Changes in lifestyle factors after the disaster were associated with the frequency of laughter in the evacuation zone. Future longitudinal studies are needed to examine what factors can increase the frequency of laughter.

     

Cytochemical analysis of storage materials in cultured skin fibroblasts from patients with I-cell disease

BACKGROUND: In cultured fibroblasts from I-cell disease patients the transport of many lysosomal enzymes is defective, and affected cells contain inclusion bodies filled with undegraded substrates. However, the contents of these inclusion bodies have not been well characterized yet. We attempted to identify accumulated substances in cultured I-cell disease fibroblasts cytochemically. METHODS: Cultured fibroblasts from I-cell disease patients were double-stained with a monoclonal antibody to lysosome-associated membrane protein-1 (LAMP-1) and that to GM2 ganglioside, or a series of lectins that specifically bind to sugar moieties. RESULTS: The patients' cells were granularly stained with the antibody to GM2 ganglioside and the lectins including Maakia amurensis, Datura stramonium, and concanavalin A. Their localization was coincident with that of LAMP-1. CONCLUSIONS: GM2 ganglioside and various kinds of glycoconjugates having sialic acidalpha2-3galactose, galactosebeta1-4N-acetylglucosamine and mannose residues accumulate in enlarged lysosomes in I-cell disease fibroblasts.