Cryptotanshinone is a candidate therapeutic agent for interstitial lung disease associated with a BRICHOS-domain mutation of SFTPC.

Interstitial lung disease (ILD) represents a large group of diseases characterized by chronic inflammation and fibrosis of the lungs, for which therapeutic options are limited. Among several causative genes of familial ILD with autosomal dominant inheritance, the mutations in the BRICHOS domain of SFTPC cause protein accumulation and endoplasmic reticulum stress by misfolding its proprotein. Through a screening system using these two phenotypes in HEK293 cells and evaluation using alveolar epithelial type 2 (AT2) cells differentiated from patient-derived induced pluripotent stem cells (iPSCs), we identified Cryptotanshinone (CPT) as a potential therapeutic agent for ILD. CPT decreased cell death induced by mutant SFTPC overexpression in A549 and HEK293 cells and ameliorated the bleomycin-induced contraction of the matrix in fibroblast-dependent alveolar organoids derived from iPSCs with SFTPC mutation. CPT and this screening strategy can apply to abnormal protein-folding-associated ILD and other protein-misfolding diseases.

Branchpoints as potential targets of exon-skipping therapies for genetic disorders.

Fukutin (FKTN) c.647+2084G>T creates a pseudo-exon with a premature stop codon, which causes Fukuyama congenital muscular dystrophy (FCMD). We aimed to ameliorate aberrant splicing of FKTN caused by this variant. We screened compounds focusing on splicing regulation using the c.647+2084G>T splicing reporter and discovered that the branchpoint, which is essential for splicing reactions, could be a potential therapeutic target. To confirm the effectiveness of branchpoints as targets for exon skipping, we designed branchpoint-targeted antisense oligonucleotides (BP-AONs). This restored normal FKTN mRNA and protein production in FCMD patient myotubes. We identified a functional BP by detecting splicing intermediates and creating BP mutations in the FKTN reporter gene; this BP was non-redundant and sufficiently blocked by BP-AONs. Next, a BP-AON was designed for a different FCMD-causing variant, which induces pathogenic exon trapping by a common SINE-VNTR-Alu-type retrotransposon. Notably, this BP-AON also restored normal FKTN mRNA and protein production in FCMD patient myotubes. Our findings suggest that BPs could be potential targets in exon-skipping therapeutic strategies for genetic disorders.

Image diagnosis models for the oral assessment of older people using convolutional neural networks: A retrospective observational study.

AIMS: The purpose of this study was to construct a model for oral assessment using deep learning image recognition technology and to verify its accuracy. BACKGROUND: The effects of oral care on older people are significant, and the Oral Assessment Guide has been used internationally as an effective oral assessment tool in clinical practice. However, additional training, education, development of user manuals and continuous support from a dental hygienist are needed to improve the inter-rater reliability of the Oral Assessment Guide. DESIGN: A retrospective observational study. METHODS: A total of 3,201 oral images of 114 older people aged >65 years were collected from five dental-related facilities. These images were divided into six categories (lips, tongue, saliva, mucosa, gingiva, and teeth or dentures) that were evaluated by images, out of the total eight items that comprise components of the Oral Assessment Guide. Each item was classified into a rating of 1, 2 or 3. A convolutional neural network, which is a deep learning method used for image recognition, was used to construct the image recognition model. The study methods comply with the STROBE checklist. RESULTS: We constructed models with a classification accuracy of 98.8% for lips, 94.3% for tongue, 92.8% for saliva, 78.6% for mucous membranes, 93.0% for gingiva and 93.6% for teeth or dentures. CONCLUSIONS: Highly accurate diagnostic imaging models using convolutional neural networks were constructed for six items of the Oral Assessment Guide and validated. In particular, for the five items of lips, tongue, saliva, gingiva, and teeth or dentures, models with a high accuracy of over 90% were obtained. RELEVANCE TO CLINICAL PRACTICE: The model built in this study has the potential to contribute to obtain reproducibility and reliability of the ratings, to shorten the time for assessment, to collaborate with dental professionals and to be used as an educational tool.

Molecular characterisation of equine group A rotavirus, Nasuno, isolated in Tochigi Prefecture, Japan.

In this study, equine group A rotavirus (RV-A), Nasuno, isolated from foal diarrhoea in Tochigi Prefecture, Japan was characterised genetically by sequence analysis of the genome segments encoding VP4 and VP7. The nucleotide and deduced amino acid sequences revealed high homology with P[12] RV-As (94.0-99.3% and 94.9-99.4%) and G3 RV-As (86.9-99.5% and 91.1-99.4%). Nasuno was also classified into P[12] and G3 in the phylogenetic analysis of the nucleotide sequences of the genome segments encoding VP4 and VP7.

An outbreak of systemic granulomatous disease in cows with high milk yields.

Seven of 92 lactating Holstein cows on a dairy farm developed urticaria with alopecia and decreased milk production, and three of the seven died over the course of 7 to 18 days. Pathologic examination of the three cows, including the two dead and one euthanized cow, revealed that the skin, liver, spleen, kidneys, heart, salivary glands, pancreas, adrenal glands, mammary glands, lymph nodes, and trigeminal ganglia had lymphocytic to lymphogranulomatous inflammation. Inflammation predominated by lymphocytic infiltration was prominent in the heart, pancreas, mammary glands, adrenal gland, and trigeminal ganglia. Severe granulomatous inflammation with multinucleated giant cells was present in the spleen and kidneys. These lesions and their distributions were most similar to those seen in suspected cases of citrus pulp toxicosis and hairy vetch toxicosis. The outbreak of this disease resolved with the elimination of Citrus pulp from the feed. Immunohistochemical detection of lymphocytes and macrophage markers confirmed dramatic hyperplasia of CD3-positive T lymphocytes in these lesions. This strongly suggested that a type 4 hypersensitivity reaction played a role in the development of the lesions.