Exome sequencing reveals a novel TTC19 mutation in an autosomal recessive spinocerebellar ataxia patient

BACKGROUND: Spinocerebellar ataxias (SCAs) are heterogeneous diseases characterized by progressive cerebellar ataxia associated with dysarthria, oculomotor abnormalities, and mental impairment. To identify the causative gene, we performed exome sequencing on a Japanese patient clinically diagnosed with recessive SCA. METHOD: The patient is a 37-year-old Japanese woman with consanguineous parents. The head magnetic resonance imaging (MRI) showed cerebellar atrophy and T1 low/T2 high intensity at the bilateral inferior olives. Single-nucleotide polymorphism (SNP) genotyping and next-generation sequencing were performed, and the variants obtained were filtered and prioritized. RESULTS: After these manipulations, we identified a homozygous nonsense mutation of the TTC19 gene (p.Q277*). TTC19 has been reported to be a causative gene of a neurodegenerative disease in Italian and Portuguese families and to be involved in the pathogenesis of mitochondrial respiratory chain complex III (cIII) deficiency. This report is the first description of a TTC19 mutation in an Asian population. Clinical symptoms and neuroimaging are consistent with previous reports. The head MRI already showed abnormal features four years before her blood lactate and pyruvate levels were elevated. CONCLUSIONS: We should consider the genetic analysis of TTC19 when we observe such characteristic MRI abnormalities. Genes associated with mitochondrial function cause many types of SCAs; the mutation we identified should help to elucidate the pathology of these disorders

BATTLE: Genetically Engineered Strategies for Split-Tunable Allocation of Multiple Transgenes in the Nervous System

Elucidating fine architectures and functions of cellular and synaptic connections requires development of new flexible methods. Here, we created a concept called the “battle of transgenes,” based on which we generated strategies using genetically engineered battles of multiple recombinases. The strategies enabled split-tunable allocation of multiple transgenes. We demonstrated the versatility of these strategies and technologies in inducing strong and multi-sparse allocations of multiple transgenes. Furthermore, the combination of our transgenic strategy and expansion microscopy enabled three-dimensional high-resolution imaging of whole synaptic structures in the hippocampus with simultaneous visualizations of endogenous synaptic proteins. These strategies and technologies based on the battle of genes may accelerate the analysis of whole synaptic and cellular connections in diverse life science fields

BATTLE: Genetically Engineered Strategies for Split-Tunable Allocation of Multiple Transgenes in the Nervous System

Elucidating fine architectures and functions of cellular and synaptic connections requires development of new flexible methods. Here, we created a concept called the “battle of transgenes,” based on which we generated strategies using genetically engineered battles of multiple recombinases. The strategies enabled split-tunable allocation of multiple transgenes. We demonstrated the versatility of these strategies and technologies in inducing strong and multi-sparse allocations of multiple transgenes. Furthermore, the combination of our transgenic strategy and expansion microscopy enabled three-dimensional high-resolution imaging of whole synaptic structures in the hippocampus with simultaneous visualizations of endogenous synaptic proteins. These strategies and technologies based on the battle of genes may accelerate the analysis of whole synaptic and cellular connections in diverse life science fields

Bacterial nucleoid dynamics: oxidative stress response in Staphylococcus aureus

A single-molecule-imaging technique, atomic force microscopy (AFM) was applied to the analyses of the genome architecture of Staphylococcus aureus. The staphylococcal cells on a cover glass were subjected to a mild lysis procedure that had maintained the fundamental structural units in Escherichia coli. The nucleoids were found to consist of fibrous structures with diameters of 80 and 40 nm. This feature was shared with the E. coli nucleoid. However, whereas the E. coli nucleoid dynamically changed its structure to a highly compacted one towards the stationary phase, the S. aureus nucleoid never underwent such a tight compaction under a normal growth condition. Bioinformatic analysis suggested that this was attributable to the lack of IHF that regulate the expression of a nucleoid protein, Dps, required for nucleoid compaction in E. coli. On the other hand, under oxidative conditions, MrgA (a staphylococcal Dps homolog) was over-expressed and a drastic compaction of the nucleoid was detected. A knock-out mutant of the gene encoding the transcription factor (perR) constitutively expressed mrgA, and its nucleoid was compacted without the oxidative stresses. The regulatory mechanisms of Dps/MrgA expression and their biological significance were postulated in relation to the nucleoid compaction

Preparation of Lead-Free Machinable Brass dispersed MnS by Hot Extrusion of Machined Chips

The most popular recycle process of brass machined chips wastes is remelting process. However, this process is much energy consumption. On the other hand, lead has been added conventionally in order to improve the machinability of metallic materials. The development of lead-free alloy has become an important issue since lead is harmful to the human body. In this study, MnS was applied to a substitutional material of lead to improve machinability and a direct consolidation process from machined chips of 60/40 brass has been developed. Mixture of machined chips of 60/40 brass and the MnS powder of 0.1-1.0 mass% were consolidated by hot extrusion. Extrusion was conducted at 773 K and an extrusion ratio of 11.1. After the tensile test of the extruded specimen, the tensile strength of the extruded specimen with MnS and that without MnS showed a nearly equivalent value. And tensile strength of the extruded specimen with MnS particles of 106 µm in diameter was slightly higher than that of 45 µm in diameter. Elongation of the extruded specimen decreased with an increase of the MnS additive amount. The cutting resistance test and the chip observation indicated that an increase of MnS additive improved machinability

Periodontal Tissue as a Biomaterial for Hard-Tissue Regeneration following bmp-2 Gene Transfer

The application of periodontal tissue in regenerative medicine has gained increasing interest since it has a high potential to induce hard-tissue regeneration, and is easy to handle and graft to other areas of the oral cavity or tissues. Additionally, bone morphogenetic protein-2 (BMP-2) has a high potential to induce the differentiation of mesenchymal stem cells into osteogenic cells. We previously developed a system for a gene transfer to the periodontal tissues in animal models. In this study, we aimed to reveal the potential and efficiency of periodontal tissue as a biomaterial for hard-tissue regeneration following a bmp-2 gene transfer. A non-viral expression vector carrying bmp-2 was injected into the palate of the periodontal tissues of Wistar rats, followed by electroporation. The periodontal tissues were analyzed through bone morphometric analyses, including mineral apposition rate (MAR) determination and collagen micro-arrangement, which is a bone quality parameter, before and after a gene transfer. The MAR was significantly higher 3–6 d after the gene transfer than that before the gene transfer. Collagen orientation was normally maintained even after the bmp-2 gene transfer, suggesting that the bmp-2 gene transfer has no adverse effects on bone quality. Our results suggest that periodontal tissue electroporated with bmp-2 could be a novel biomaterial candidate for hard-tissue regeneration therapy.Kawai M.Y., Ozasa R., Ishimoto T., et al. Periodontal Tissue as a Biomaterial for Hard-Tissue Regeneration following bmp-2 Gene Transfer. Materials, 15, 3, 993. https://doi.org/10.3390/ma15030993

体育科における授業改善のための試行(1) : フィードバック情報の活用

本研究は，小学校の体育授業において，授業観察の結果で得られた情報を授業者にフィードバックし，それが次の授業実践の改善に向けてどのように機能しているかを確かめる事例的試行の第1報である。本論文は，教師の教授技能の一つである学習者との「相互作用」に着目し，授業中の教師（現職）の学習者に向けられた助言内容を量的・質的の両面から分析した結果の情報（フィードバック情報）が，次の授業実践においてどのように機能したかを授業記錦の定量的分析及び学習者の授業評価をもとに検証しようと試みたものである。その結果，教師の「肯定的言葉かけ」や「めあてのもたせ方」などに有効に機能していることがわかった。This study aimed to clarity how some informations from the resrlts of instructional observations effected to improve next instruction in physical education. Two instructions at primary school were chosen for this study. In this study, interaction between a teacher and learners was focused on as one of teaching skills. Through analysis of instructional observation, the interactions were picked up from the view points of qualitative and quantitative. And next, the results of former analysis were given to the teacher as informations (feedback information) for purposes of improving next instruction. From these analysis it was concluded that the feedback informations were useful for improving teacher\u27s positive talk and method to let learners know purposes