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52 result(s) for "Uchida, Koki"
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Exploring New Functional Aspects of HTLV-1 RNA-Binding Protein Rex: How Does Rex Control Viral Replication?
After integration to the human genome as a provirus, human T-cell leukemia virus type 1 (HTLV-1) utilizes host T cell gene expression machinery for viral replication. The viral RNA-binding protein, Rex, is known to transport unspliced/incompletely spliced viral mRNAs encoding viral structural proteins out of the nucleus to enhance virus particle formation. However, the detailed mechanism of how Rex avoids extra splicing of unspliced/incompletely spliced viral mRNAs and stabilizes them for effective translation is still unclear. To elucidate the underlying molecular mechanism of Rex function, we comprehensively analyzed the changes in gene expression and splicing patterns in Rex-overexpressing T cells. In addition, we identified 81 human proteins interacting with Rex, involved in transcription, splicing, translation, and mRNA quality control. In particular, Rex interacts with NONO and SFPQ, which play important roles in the regulation of transcription and splicing. Accordingly, expression profiles and splicing patterns of a wide variety of genes are significantly changed in Rex-expressing T cells. Especially, the level of vPD-L1 mRNA that lacks the part of exon 4, thus encodes soluble PD-L1 was significantly increased in Rex-expressing cells. Overall, by integrated analysis of these three datasets, we showed for the first time that Rex intervenes the host gene expression machinery throughout the pathway, probably to escort viral unstable mRNAs from transcription (start) to translation (end). Upon exerting its function, Rex may alter the expression level and splicing patterns of various genes, thus influencing the phenotype of the host cell.
Cell culture and genetic transfection methods for the Japanese scallop, Patinopecten yessoensis
Cell cultures can simplify assays of biological phenomena; therefore, cell culture systems have been established for many species, even invertebrates. However, there are few primary culture systems from marine invertebrates that can be maintained long term. The Japanese scallop, Patinopecten yessoensis, is a marine bivalve. Cell culture systems for the scallop have only been established for a few organ‐derived cell types and for embryonic cells. We developed a primary culture system for cells from male and female scallop gonads, hepatopancreas, and adductor muscle by utilizing culture conditions closer to those in nature, with regard to temperature, osmolarity, and nutrition. Primary cultured female gonadal cells were maintained for more than 1 month and had potential for proliferation. Furthermore, a genetic transfection system was attempted using a scallop‐derived promoter and a lipofection reagent. GFP‐positive cells were detected in the attempt. These technical developments would promote our understanding of biochemical mechanisms in scallops as well as providing clues for establishment of immortalized molluscan cell lines. We developed a primary culture system for female and male gonad‐, hepatopancreas‐, and adductor muscle‐derived cells from Japanese scallop, Patinopecten yessoensis. In the female gonadal cell culture, we succeeded in maintaining the cultured cells for at least one month and the culture had potential for cell proliferation. We attempted transfection of female gonadal cells with scallop‐derived promoters and lipofection reagent.
Nestin Forms a Flexible Cytoskeleton by Means of a Huge Tail Domain That Is Reversibly Stretched and Contracted by Weak Forces
Nestin is a type VI intermediate filament protein and a well-known neural stem cell marker. It is also expressed in high-grade cancer cells, forming copolymerized filaments with vimentin. We previously showed that nestin inhibits the binding of vimentin’s tail domain to actin filaments (AFs) by steric hindrance through its large nestin tail domain (NTD), thereby increasing three-dimensional cytoskeleton network mobility, enhancing cell flexibility, and promoting cancer progression. Further, we found that nestin itself stably binds to AFs via the NTD. We therefore hypothesized that the NTD may form a flexible cytoskeletal structure by extending with weak force. In vitro tensile tests using atomic force microscopy were performed to assess the mechanical properties of NTDs. The C-terminus of the NTD bound AFs by bringing the AFM tip modified with the NTD into contact with the AFs on the substrate. NTDs were elongated to approximately 80% of their maximum length at weak forces < 150 pN. Repeated tensile tests revealed that the NTD refolded quickly and behaved like a soft elastic material. We speculate that nestin stably binds AFs, and the NTD extends with weak force, contracting quickly upon load release. Thereby, nestin would absorb mechanical load and maintain cytoskeletal integrity.
Bottom‐up Synthesis of Nanosheets at Various Interfaces
Nanostructured materials with high aspect ratios have been widely studied for their unique properties. In particular, nanosheets have safety, dispersibility, and nanosized effects, and nanosheets with exceptionally small thicknesses exhibit unique properties. For non‐exfoliable materials, the bottom‐up nanosheet growth using various interfaces as templates have been investigated. This review article presents the synthesis of nanosheets at the interfaces and layered structure; it explains the features of each interface type, its advantages, and its uniqueness. The interfaces work as templates for nanosheet synthesis. We can easily use the liquid‐liquid and gas‐liquid interfaces as the templates; however, the thickness of nanosheets usually becomes thick because it allows materials to grow in thickness. The solid‐gas and solid‐liquid interfaces can prevent nanosheets from growing in thickness. However, the removal of template solids is required after the synthesis. The layered structures of various materials provide two‐dimensional reaction fields between the layers. These methods have high versatility, and the nanosheets synthesized by these methods are thin. Finally, this review examines the key challenges and opportunities associated with scalable nanosheet synthesis methods for industrial production.
Sm-Co-based amorphous alloy films for zero-field operation of transverse thermoelectric generation
Transverse thermoelectric generation using magnetic materials is essential to develop active thermal engineering technologies, for which the improvements of not only the thermoelectric output but also applicability and versatility are required. In this study, using combinatorial material science and lock-in thermography technique, we have systematically investigated the transverse thermoelectric performance of Sm-Co-based alloy films. The high-throughput material investigation revealed the best Sm-Co-based alloys with the large anomalous Nernst effect (ANE) as well as the anomalous Ettingshausen effect (AEE). In addition to ANE/AEE, we discovered unique and superior material properties in these alloys: the amorphous structure, low thermal conductivity, and large in-plane coercivity and remanent magnetization. These properties make it advantageous over conventional materials to realize heat flux sensing applications based on ANE, as our Sm-Co-based films can generate thermoelectric output without an external magnetic field. Importantly, the amorphous nature enables the fabrication of these films on various substrates including flexible sheets, making the large-scale and low-cost manufacturing easier. Our demonstration will provide a pathway to develop flexible transverse thermoelectric devices for smart thermal management.
Podocyte autophagy is associated with foot process effacement and proteinuria in patients with minimal change nephrotic syndrome
Autophagy is a cellular mechanism involved in the bulk degradation of proteins and turnover of organelle. Several studies have shown the significance of autophagy of the renal tubular epithelium in rodent models of tubulointerstitial disorder. However, the role of autophagy in the regulation of human glomerular diseases is largely unknown. The current study aimed to demonstrate morphological evidence of autophagy and its association with the ultrastructural changes of podocytes and clinical data in patients with idiopathic nephrotic syndrome, a disease in which patients exhibit podocyte injury. The study population included 95 patients, including patients with glomerular disease (minimal change nephrotic syndrome [MCNS], n = 41; idiopathic membranous nephropathy [IMN], n = 37) and 17 control subjects who underwent percutaneous renal biopsy. The number of autophagic vacuoles and the grade of foot process effacement (FPE) in podocytes were examined by electron microscopy (EM). The relationships among the expression of autophagic vacuoles, the grade of FPE, and the clinical data were determined. Autophagic vacuoles were mainly detected in podocytes by EM. The microtubule-associated protein 1 light chain 3 (LC3)-positive area was co-localized with the Wilms tumor 1 (WT1)-positive area on immunofluorescence microscopy, which suggested that autophagy occurred in the podocytes of patients with MCNS. The number of autophagic vacuoles in the podocytes was significantly correlated with the podocyte FPE score (r = -0.443, p = 0.004), the amount of proteinuria (r = 0.334, p = 0.033), and the level of serum albumin (r = -0.317, p = 0.043) in patients with MCNS. The FPE score was a significant determinant for autophagy after adjusting for the age in a multiple regression analysis in MCNS patients (p = 0.0456). However, such correlations were not observed in patients with IMN or in control subjects. In conclusion, the results indicated that the autophagy of podocytes is associated with FPE and severe proteinuria in patients with MCNS. The mechanisms underlying the activation of autophagy in association with FPE in podocytes should be further investigated in order to elucidate the pathophysiology of MCNS.
Large spin-Hall effect in non-equilibrium binary copper alloys beyond the solubility limit
Non-magnetic materials exhibiting large spin-Hall effect (SHE) are eagerly desired for high-performance spintronic devices. Here, we report that non-equilibrium Cu-Ir binary alloys with compositions beyond the solubility limit are candidates as spin-Hall materials, even though Cu and Ir do not exhibit remarkable SHE themselves. Thanks to non-equilibrium thin film fabrication, the Cu-Ir binary alloys are obtained over a wide composition range even though they are thermodynamically unstable in bulk form. We investigate the SHE of Cu-Ir by exploiting a combinatorial technique based on spin Peltier imaging, and find that the optimum Ir concentration for enhancing SHE is around 25 at.%. We achieve a large spin-Hall angle of 6.29 ± 0.19% for Cu76Ir24. In contrast to Cu-Ir, non-equilibrium Cu-Bi binary alloys do not show remarkable SHE. Our discovery opens a new direction for the exploration of spin-Hall materials.Materials with a large spin-Hall effect are highly desirable for spintronic devices. Here, non-equilibrium thin film synthesis is used to fabricate copper-iridium binary alloys beyond their solubility limit, achieving a large spin-Hall angle of approximately 6% in Cu76Ir24.
Efficient Manipulation of Magnetic Domain Wall by Dual Spin‐Orbit Torque in Synthetic Antiferromagnets
Current‐induced domain‐wall motion (CIDWM) in a synthetic antiferromagnet is a key phenomenon for developing potential high‐density‐packed magnetic domain‐wall memory with fast operation. Here, CIDWM is reported in the antiferromagnetically‐coupled two Co layers through the Ir interlayer sandwiched by the two Pt layers: Pt/Co/Ir/Co/Pt. The top and bottom Pt layers play a role for generating the spin current coming from the spin Hall effect, which gives rise to the dual spin‐orbit torque (SOT) acting on the perpendicular magnetizations of the Co layers. Although a simple argument would predict that SOTs from top and bottom Pt layers cancel each other out, the dual SOT nucleates a reversed magnetic domain and drives the CIDWM effectively at current density of the order of 1011 A m−2. This study also examines the effect of antisymmetric interlayer exchange coupling (AIEC) on CIDWM. A positive correlation between the magnitude of AIEC and the domain wall velocity is found, whereas the current density required for nucleating the reversed domain shows a negative correlation with the magnitude of AIEC. These facts suggest that the existence of AIEC improves the performance of CIDWM. The present results provide a new avenue to design SOT domain wall devices based on a synthetic antiferromagnet. Dual spin‐orbit torque coming from two Pt layers effectively drives the magnetic domain wall motion (DWM) in a synthetic antiferromagnet consisting of two Co ferromagnetic layers coupled with interlayer exchange coupling (IEC). In addition, the antisymmetric version of IEC leads to low current density for nucleating DW and fast velocity of DWM, improving the performance of magnetic DW devices.
Deep convolutional neural networks for automated scoring of pentagon copying test results
This study aims to investigate the accuracy of a fine-tuned deep convolutional neural network (CNN) for evaluating responses to the pentagon copying test (PCT). To develop a CNN that could classify PCT images, we fine-tuned and compared the pre-trained CNNs (GoogLeNet, VGG-16, ResNet-50, Inception-v3). To collate our training dataset, we collected 1006 correct PCT images and 758 incorrect PCT images drawn on a test sheet by dementia suspected patients at the Osaka City Kosaiin Hospital between April 2009 and December 2012. For a validation dataset, we collected PCT images from consecutive patients treated at the facility in April 2020. We examined the ability of the CNN to detect correct PCT images using a validation dataset. For a validation dataset, we collected PCT images (correct, 41; incorrect, 16) from 57 patients. In the validation testing for an ability to detect correct PCT images, the fine-tuned GoogLeNet CNN achieved an area under the receiver operating characteristic curve of 0.931 (95% confidence interval 0.853–1.000). These findings indicate that our fine-tuned CNN is a useful method for automatically evaluating PCT images. The use of CNN-based automatic scoring of PCT can potentially reduce the burden on assessors in screening for dementia.