Flap endonuclease 1 is involved in cccDNA formation in the hepatitis B virus

金沢大学医薬保健研究域医学系Hepatitis B virus (HBV) is one of the major etiological pathogens for liver cirrhosis and hepatocellular carcinoma. Chronic HBV infection is a key factor in these severe liver diseases. During infection, HBV forms a nuclear viral episome in the form of covalently closed circular DNA (cccDNA). Current therapies are not able to efficiently eliminate cccDNA from infected hepatocytes. cccDNA is a master template for viral replication that is formed by the conversion of its precursor, relaxed circular DNA (rcDNA). However, the host factors critical for cccDNA formation remain to be determined. Here, we assessed whether one potential host factor, flap structure-specific endonuclease 1 (FEN1), is involved in cleavage of the flap-like structure in rcDNA. In a cell culture HBV model (Hep38.7-Tet), expression and activity of FEN1 were reduced by siRNA, shRNA, CRISPR/Cas9-mediated genome editing, and a FEN1 inhibitor. These reductions in FEN1 expression and activity did not affect nucleocapsid DNA (NC-DNA) production, but did reduce cccDNA levels in Hep38.7-Tet cells. Exogenous overexpression of wild-type FEN1 rescued the reduced cccDNA production in FEN1-depleted Hep38.7-Tet cells. Anti-FEN1 immunoprecipitation revealed the binding of FEN1 to HBV DNA. An in vitro FEN activity assay demonstrated cleavage of 5′-flap from a synthesized HBV DNA substrate. Furthermore, cccDNA was generated in vitro when purified rcDNA was incubated with recombinant FEN1, DNA polymerase, and DNA ligase. Importantly, FEN1 was required for the in vitro cccDNA formation assay. These results demonstrate that FEN1 is involved in HBV cccDNA formation in cell culture system, and that FEN1, DNA polymerase, and ligase activities are sufficient to convert rcDNA into cccDNA in vitro

Generation of electromagnetic waves from 0.3 to 1.6 terahertz with a high-T-c superconducting Bi2Sr2CaCu2O8+delta intrinsic Josephson junction emitter

To obtain higher power P and frequency f emissions from the intrinsic Josephson junctions in a high-T-c superconducting Bi2Sr2CaCu2O8+delta single crystal, we embedded a rectangular stand-alone mesa of that material in a sandwich structure to allow for efficient heat exhaust. By varying the current-voltage (I-V) bias conditions and the bath temperature T-b, f is tunable from 0.3 to 1.6 THz. The maximum P of a few tens of mu W, an order of magnitude greater than from previous devices, was found at T-b similar to 55K on an inner I-V branch at the TM(1,0) cavity resonance mode frequency. The highest f of 1.6 THz was found at T-b = 10K on an inner I-V branch, but away from cavity resonance frequencies. A possible explanation is presented

The whole blood transcriptional regulation landscape in 465 COVID-19 infected samples from Japan COVID-19 Task Force

「コロナ制圧タスクフォース」COVID-19患者由来の血液細胞における遺伝子発現の網羅的解析 --重症度に応じた遺伝子発現の変化には、ヒトゲノム配列の個人差が影響する--. 京都大学プレスリリース. 2022-08-23.Coronavirus disease 2019 (COVID-19) is a recently-emerged infectious disease that has caused millions of deaths, where comprehensive understanding of disease mechanisms is still unestablished. In particular, studies of gene expression dynamics and regulation landscape in COVID-19 infected individuals are limited. Here, we report on a thorough analysis of whole blood RNA-seq data from 465 genotyped samples from the Japan COVID-19 Task Force, including 359 severe and 106 non-severe COVID-19 cases. We discover 1169 putative causal expression quantitative trait loci (eQTLs) including 34 possible colocalizations with biobank fine-mapping results of hematopoietic traits in a Japanese population, 1549 putative causal splice QTLs (sQTLs; e.g. two independent sQTLs at TOR1AIP1), as well as biologically interpretable trans-eQTL examples (e.g., REST and STING1), all fine-mapped at single variant resolution. We perform differential gene expression analysis to elucidate 198 genes with increased expression in severe COVID-19 cases and enriched for innate immune-related functions. Finally, we evaluate the limited but non-zero effect of COVID-19 phenotype on eQTL discovery, and highlight the presence of COVID-19 severity-interaction eQTLs (ieQTLs; e.g., CLEC4C and MYBL2). Our study provides a comprehensive catalog of whole blood regulatory variants in Japanese, as well as a reference for transcriptional landscapes in response to COVID-19 infection

Visualizing sphere-contacting areas on automobile parts for ECE inspection

To satisfy safety regulations of Economic Commission for Europe (ECE), the surface regions of automobile parts must have a sufficient degree of roundness if there is any chance that they could contact a sphere of 50.0 mm radius (exterior parts) or 82.5 mm radius (interior parts). In this paper, a new offset-based method is developed to automatically detect the possible sphere-contacting shape of such parts. A polyhedral model that precisely approximates the part shape is given as input, and the offset shape of the model is obtained as the Boolean union of the expanded shapes of all surface triangles. We adopt a triple-dexel representation of the 3D model to enable stable and precise Boolean union computations. To accelerate the dexel operations in these Boolean computations, a new parallel processing method with a pseudo-list structure and axis-aligned bounding box is developed. The possible sphere-contacting shape of the part surface is then extracted from the offset shape as a set of points or a set of polygons

Visualization of the entire differentiation process of murine M cells : suppression of their maturation in cecal patches

The microfold (M) cell residing in the follicle-associated epithelium is a specialized epithelial cell that initiates mucosal immune responses by sampling lumina! antigens. The differentiation process of M cells remains unclear due to limitations of analytical methods. Here we found that M cells were classified into two functionally different subtypes based on the expression of Glycoprotein 2 (GP2) by newly developed image cytometric analysis. GP2-high M cells actively took up luminal microbeads, whereas GP2-negative or low cells scarcely ingested them, even though both subsets equally expressed the other M-cell signature genes, suggesting that GP2-high M cells represent functionally mature M cells. Further, the GP2-high mature M cells were abundant in Peyer's patch but sparse in the cecal patch: this was most likely due to a decrease in the nuclear translocation of RelB, a downstream transcription factor for the receptor activator of nuclear factor-kappa B signaling. Given that murine cecum contains a protrusion of beneficial commensals, the restriction of M-cell activity might contribute to preventing the onset of any excessive immune response to the commensals through decelerating the M-cell-dependent uptake of microorganisms

Styles of cognition toward the Depersonalization/Derealization in undergrade students : A free-description questionnaire study

The purpose of this study is to clarify how the undergraduate students cognize the DepersonalizationDerealization (DP/DR). DP/DR is a subjective feeling of unreality of the self and the outside world. In spite of the feeling of unreality, one keeps reality testing under DP/DDR. In this study, Five question items, which were selected from \u22The Cambridge Depersonalisation Scale-Japanese Edition (Tanabe, 2004)\u22, were used and administered to 22 undergraduate students. 131 items were answered and classified with K-J method. The results of the classifications are 5 big categories, 10 middle categories and 25 small categories. Although \u22negative cognition\u22 holds more than 70%, “positive cognition2 holds 9.2%. Shimoyama (2001)mentions \u22Support of Clinical psychology is to support the story where the human lives with holding illness.\u22 Therefore, it is important to support this \u22positive cognition\u22 from the point of clinical psychology

自由記述式の質問紙による研究

The purpose of this study is to clarify how the undergraduate students cognize the Depersonalization/Derealization (DP/DR). DP/DR is a subjective feeling of unreality of the self and the outside world. In spite of the feeling of unreality, one keeps reality testing under DP/DDR. In this study, Five question items, which were selected from "The Cambridge Depersonalisation Scale-Japanese Edition (Tanabe, 2004)", were used and administered to 22 undergraduate students. 131 items were answered and classified with K-J method. The results of the classifications are 5 big categories, 10 middle categories and 25 small categories. Although "negative cognition" holds more than 70%, “positive cognition2 holds 9.2%. Shimoyama (2001)mentions "Support of Clinical psychology is to support the story where the human lives with holding illness." Therefore, it is important to support this "positive cognition" from the point of clinical psychology