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Acute disseminated encephalomyelitis is an immune-mediated inflammatory demyelinating disease of the central nervous system that can occur during or after infections. A 2-year-old female was hospitalized in an unconsciousness state with fever. She had suffered from 5 days of hand-foot-mouth disease until 2 days before admission. Magnetic resonance imaging disclosed high-intensity lesions at the cerebrum and cerebellum but not at the brainstem and spinal cord, confirming the diagnosis of acute disseminated encephalomyelitis. Her illness was remarkably improved after the start of steroid therapy and she could recover without any sequele. The genetic analysis of the genome sequence revealed that the isolated strain was enterovirus 71. The phylogenetic analysis with the strains obtained from fatal brainstem encephalitis suggests an association between the genotypes and the neurological virulency. [ABSTRACT FROM AUTHOR]

Many recombinant proteins have been used as drugs; however, human proteins expressed using heterologous hosts are often insoluble. To obtain correctly folded active proteins, many optimizations of expression have been attempted but usually are found to be applicable only for specific targets. Interleukin-18 (IL-18) has a key role in many severe disorders including autoimmune diseases, and therapeutic approaches using IL-18 have been reported. However, production of IL-18 in Escherichia coli resulted in extensive inclusion body formation and previous conventional screenings of expression conditions could obtain only a condition with a low yield. To address the problem, we applied a folding reporter system using green fluorescent protein (GFP) for screening of the expression conditions for hIL-18. The established system efficiently screened many conditions, and optimized conditions for the expression of hIL-18 significantly enhanced the final yield of the active protein. Systematic screening using a GFP reporter system could be applied for the production of other proteins and in other organisms. [Copyright &y& Elsevier]

Background: Human interferon-gamma (hIFN-γ) is produced by lymphocytes and has a variety of biological properties. Measurement of hIFN-γ is widely used for various immunological responses for allergic or autoimmune diseases. Enzyme-linked immunosorbent assay (ELISA) is an established immunoassay used to quantify cellular metabolites or cytokines. ELISA requires many incubation and wash steps and is not practically suitable for screening large numbers of samples. Methods: We have developed a fluorescence-linked immunosorbent assay (FLISA) method for the detection of hIFN-γ. We measured the 50% inhibitory concentration (IC50) value of the hIFN-γ production by interleukin (IL)-18 binding protein and anti-IL-18 monoclonal antibody. The IC50 described by FLISA was compared with that by ELISA. Results: We developed a new system for measuring hIFN-γ using Allophycocyanine (APC) fluorescent protein and compared it with the previous method using Cy5.5. The proposed FLISA had a smaller coefficient of variation than ELISA, and the means of coefficient of variation using the same samples measured by ELISA and FLISA were, respectively, 11.1% and 3.8%, suggesting that the edge effect often giving non-specific results may be smaller in FLISA than in ELISA. Conclusions: The improved FLISA system proposed is ideally suited for efficient measurements of hIFN-γ. This homogeneous and multiplex method will be a powerful tool for high throughput screening for drug discovery research. [ABSTRACT FROM AUTHOR]

Background: The aim of this study was to investigate the prediction of postnatal prognosis using fetal and perinatal data in patients with primary congenital dilated cardiomyopathy (PCDCM), and to estimate the incidence of this disease. Methods: We examined correlations between fetal or perinatal data and postnatal clinical course in a multicenter retrospective study of eight patients with PCDCM. Incidence was calculated in a population‐based study. Results: All patients developed heart failure at a median of 8 days (range, 0–43 days), and six patients died or required extracorporeal artificial heart therapy at a median of 67 days (range, 0–92 days). The cardiothoracic area ratio from fetal echocardiography, the Apgar score, and the standard deviation of birth weight correlated significantly with the date at onset of heart failure. However, no data correlated with survival. Cumulative incidence of PCDCM was calculated as 1.21 per 100 000 total births (95% confidence interval, 0.37 to 2.06). Conclusions: Primary congenital dilated cardiomyopathy has a poor prognosis, but cardiothoracic area ratio from fetal echocardiography, body weight at birth, and Apgar score correlate with the timing of the onset of heart failure, and these indicators might therefore be useful for peri‐ and postnatal management. [ABSTRACT FROM AUTHOR]

Abstract: Van der Knaap disease, characterized by megalencephalic leukoencephalopathy and subcortical cysts, is a rare and recently defined condition. We discuss here the MR image (MRI) and MR spectroscopy (MRS) features in a 30-year-old man with S93L homozygous mutation in the MLC1 gene. MRI demonstrated high intensity diffuse white matter with T2-weighted image and subcortical cysts in the parietal and temporal lobes and MRS showed mildly reduced N-acetylaspartate (NAA) in areas of severe T2 elongation with a long TE sequence. A peak of lactate/lipid was indicated at a chemical shift of 1.3ppm with a short TE sequence. The peak for myo-inositol was normal in areas of severe and mild T2 elongation with short TE MRS. These findings suggest that demyelination progresses slowly in van der Knaap disease and that MRS with long and short TE is useful for the evaluation of neural metabolization associated with van der Knaap disease. [Copyright &y& Elsevier]

Sotos syndrome is a well-known anomaly syndrome characterized by overgrowth, characteristic facial gestalt, and developmental delay, and haploinsufficiency of the NSD1 gene has been revealed as one of the major genetic causes. However, there have been only a few reports on neuroradiologic findings by computed Sotos syndrome is an anomaly syndrome characterized by overgrowth, characteristic facial gestalt, and developmental delay. Recently, haploinsufficiency of the NSD1 gene was revealed as one of the major genetic causes of Sotos syndrome. From the first report by Sotos et al, more than 200 cases have been reported to date, but there have been only a few reports on neuroradiologic findings by computed tomography (CT) or magnetic resonance imaging (MRI). Moreover, functional examination by using single-photon emission computed tomography (SPECT) or magnetic resonance spectroscopy has not been reported. We report here the neuroradiologic findings of three patients with Sotos syndrome examined by MRI, SPECT, and magnetic resonance spectroscopy, suggesting an immaturity in frontal brain function. [ABSTRACT FROM AUTHOR]

Atopic disorders, such as asthma, eczema and rhinitis, develop due to the interactions between genetic and environmental factors. Atopy is characterized by enhanced IgE responses to environmental antigens. The production of IgE is upregulated by Th2 cytokines, in particular interleukin (IL)-4, and downregulated by Th1 cytokines, in particular interferon (IFN)-γ. In the present review, we present the genetic factors responsible for IgE production and genetic defects in the downregulation (brake) of IgE production, especially in terms of IL-12 and IL-18 signaling, mutations of the IL-12 receptor β2 chain gene and mutations of the IL-18 receptor α chain gene in atopy. Moreover, we newly present a genetic classification of atopy. There are four categories of genes that control the expression of allergic disorders, which include: (i) antigen recognition; (ii) IgE production (downregulation = brake; and upregulation); (iii) the production and release of mediators; and (iv) events on target organs. In the near future, this genetic classification will facilitate the development of tailor-made treatment. [ABSTRACT FROM AUTHOR]

Human interleukin-18 (hIL-18), initially cloned as an IFN-γ-inducing factor, has a key role in many inflammatory diseases. We have previously developed a high production system for correctly folded active hIL-18 protein, leading to the revelation of the 3D-structure and the receptor binding mode. These findings can strongly indicate the experimental and medical applications of IL-18; however, the recombinant protein is prone to be inactivated forming multimers. Recently, therapeutic approaches using recombinant IL-18 have shown the effectiveness for treatment of cancer; indicating the necessity of a more stable protein for therapy with intertrial reliability. Here we have generated a highly stable hIL-18 with replacement of cysteine by serine based on the tertiary structure and the binding mechanism, retaining the biological activity. Similar rational designs can be applied to develop new therapeutic molecules of other cytokines. [Copyright &y& Elsevier]

Human interleukin-18 (hIL-18), originally known as an IFN-γ-inducing factor, is a recently cloned cytokine that is secreted by Kupffer cells of the liver and by stimulated macrophages. We have previously established a method of expression and purification of IL-18. The yield however remains low and the insufficient expression of a heterologous protein could be due to skewed codon usage between the expression host and the cDNA donor. The sequence of mature hIL-18 has 37 a.a. rare codons for Escherichia coli in a total of 157 a.a. To overcome this problem, gene synthesis was performed with optimized codons for the expression host E. coli. The final yield of the hIL-18 protein with optimized codons was about five times higher than the yield with the native sequence. Using a minimal medium, this system produces large quantities of labeled proteins that can be used in NMR analysis. Our simple and efficient production system can be applied to the production of other cytokines for new structural and therapeutic use. [Copyright &y& Elsevier]

Interleukin-18 (IL-18), a cytokine formerly known as interferon-?- (IFN-?-) inducing factor, has pleiotropic immunoregulatory functions, including augmentation of IFN-? production, Fas-mediated cytotoxicity and developmental regulation of T-lymphocyte helper type I. We determined the solution structure of IL-18 as a first step toward understanding its receptor activation mechanism. It folds into a ß-trefoil structure that resembles that of IL-1. Extensive mutagenesis revealed the presence of three sites that are important for receptor activation: two serve as binding sites for IL-18 receptor a (IL-18Ra), located at positions similar to those of IL-1 for IL-1 receptor type I (IL-1RI), whereas the third site may be involved in IL-18 receptor ß (IL-18Rß) binding. The structure and mutagenesis data provide a basis for understanding the IL-18-induced heterodimerization of receptor subunits, which is necessary for receptor activation. [ABSTRACT FROM AUTHOR]