Eradication of Hepatitis C Virus Subgenomic Replicon by Interferon Results in Aberrant Retinol-Related Protein Expression

Hepatitis C virus (HCV) infection induces several changes in hepatocytes, such as oxidative stress, steatosis, and hepatocarcinogenesis. Although considerable progress has been made during recent years, the mechanisms underlying these functions remain unclear. We employed proteomic techniques in HCV replicon-harboring cells to determine the effects of HCV replication on host-cell protein expression. We examined two-dimensional electrophoresis (2-DE) and mass spectrometry to compare and identify differentially expressed proteins between HCV subgenomic replicon-harboring cells and their “cured” cells. One of the identified proteins was confirmed using enzyme-linked immunosorbent assay (ELISA) and Western blot analysis. Full-length HCV genome RNA replicating and cured cells were also assessed using ELISA. Replicon-harboring cells showed higher expression of retinal dehydrogenase 1 (RALDH-1), which converts retinol to retinoic acid, and the cured cells showed higher expression of retinol-binding protein (RBP), which transports retinol from the liver to target tissues. The alteration in RBP expression was also confirmed by ELISA and Western blot analysis. We conclude that protein expression profiling demonstrated that HCV replicon eradication affected retinol-related protein expression

Structural basis for potent inhibition of d-amino acid oxidase by thiophene carboxylic acids

A series of thiophene-2-carboxylic acids and thiophene-3-carboxylic acids were identified as a new class of DAO inhibitors. Structure-activity relationship (SAR) studies revealed that small substituents are well-tolerated on the thiophene ring of both the 2-carboxylic acid and 3-carboxylic acid scaffolds. Crystal structures of human DAO in complex with potent thiophene carboxylic acids revealed that Tyr224 was tightly stacked with the thiophene ring of the inhibitors, resulting in the disappearance of the secondary pocket observed with other DAO inhibitors. Molecular dynamics simulations of the complex revealed that Tyr224 preferred the stacked conformation irrespective of whether Tyr224 was stacked or not in the initial state of the simulations. MM/GBSA indicated a substantial hydrophobic interaction between Tyr244 and the thiophene-based inhibitor. In addition, the active site was tightly closed with an extensive network of hydrogen bonds including those from Tyr224 in the stacked conformation. The introduction of a large branched side chain to the thiophene ring markedly decreased potency. These results are in marked contrast to other DAO inhibitors that can gain potency with a branched side chain extending to the secondary pocket due to Tyr224 repositioning. These insights should be of particular importance in future efforts to optimize DAO inhibitors with novel scaffolds

P219L DAO alters ligand binding and catalytic efficiency

Human D-amino acid oxidase (DAO) is a flavoenzyme that is implicated in neurodegenerative diseases. We investigated the impact of replacement of proline with leucine at position 219 (P219L) in the active site lid of human DAO on the structural and enzymatic properties, because porcine DAO contains leucine at the corresponding position. The turnover numbers (kcat) of P219L were unchanged, but its Km values decreased compared to wild-type, leading to an increase in the catalytic efficiency (kcat/Km). Moreover, benzoate inhibits P219L with lower Ki value (0.7-0.9 μM) compared to wild-type (1.2-2.0 μM). Crystal structure of P219L in complex with flavin adenine dinucleotide (FAD) and benzoate at 2.25 Å resolution displayed conformational changes of the active site and lid. The distances between the H-bond-forming atoms of arginine 283 and benzoate and the relative position between the aromatic rings of tyrosine 224 and benzoate were changed in the P219L complex. Taken together, the P219L substitution leads to an increase in the catalytic efficiency and binding affinity for substrates/inhibitors due to these structural changes. Furthermore, an acetic acid was located near the adenine ring of FAD in the P219L complex. The present study provides new insights into the structure-function relationship of human DAO

日韓における出産・育児に関する慣習の比較研究

韓国（晋州）と日本（長崎県・対馬）において出産・育児に関する慣習について調査し，結果を比較した．里帰りの慣習は日本独自のものであった．出産は韓国では神聖視されており，日本では不浄視されていた．しかし，二つの地域での出産・育児の慣習は類似していた．産後の禁忌に関する慣習では，妊娠中の慣習と比較して科学的に納得のいくものが多かった．科学的知識の乏しかった時代の人々は，経験から産後の回復過程をよく観察し理解していたと思われる．誕生後はさまざまな儀式がとり行われていたが，これらは子どもの無事を願う両親や家族及び近隣社会の人々の気持ちを反映したものであった．出産・育児に関する慣習の本質は母子の安全への願いであり，それは文化の違いに関係なく存在するものであろう．The cultural background of patients who require in nursing must be considered because it has effects on the quality of nursing care. Traditional customs regarding delivery and child rearing in Korea and Japan were investigated and compared. Inherited customs in the two areas were very similar. Many taboos concerned the type of eaten food and behavior during after delivery were scientifically persuasive. Before the development of medical science, such customs whereas were the fruits of traditional wisdom. Concerning childbirth, taboos in Korea involved the element of sacredness, in Japan taboos guarded against pollution. Various rites performed after birth reflected not only the wishes of the parents and other family members, but of neighbors for the healthy growth of the child. The essence of these birth customs, regardless of differences in culture, is that people wished for the safety of the mother and child

日韓における妊娠に関する慣習の比較研究

韓国（晋州）と日本（長崎県・対馬）において妊娠に関する慣習について調査し，結果を比較した．妊婦の食生活では，韓国では82．7％，日本では16．0％が禁食を伝承していた．禁食には類感呪術というべきものが多かったが，禁食の種類は両地域に共通性がみられ，両地域とも「このタブーをおかすと胎児に何らかの影響がある」という形での伝承であった．一方，妊婦の行動上の禁忌は，韓国では79．8％，日本で34．7％が伝承していた．行動上の禁忌の内容も両地域は非常に似かよったものであった．妊婦の禁忌には明らかに迷信であるものが多く含まれていたが，禁忌の背景には医療が未発達だった時代に母子の安全を願う人々の心があったと考えられた．The cultural background of patients who require in nursing must be considered because it has effects on the quality of nursing care. Traditional customs regarding pregnancy in Korea and Japan were investigated and compared. Inherited customs in the two areas were very similar. Many taboos concerned the type of eaten food and behavior during pregnancy, and although obviously superstitions reflected underlying wishes for the safety of the mother and baby. Before the development of medical science, such customs whereas were the fruits of traditional wisdom