The Influence of Polyploidy and Genome Composition on Genomic Imprinting in Mice

Genomic imprinting is an epigenetic mechanism that switches the expression of imprinted genes involved in normal embryonic growth and development in a parent-of-origin-specific manner. Changes inDNAmethylation statuses from polyploidization are a well characterized epigenetic modification in plants. However, how changes in ploidy affect both imprinted gene expression and methylation status in mammals remains unclear. To address this, we used quantitative real time PCR to analyze expression levels of imprinted genes in mouse tetraploid fetuses. We used bisulfite sequencing to assess the methylation statuses of differentially methylated regions (DMRs) that regulate imprinted gene expression in triploid and tetraploid fetuses. The nine imprinted genes H19, Gtl2, Dlk1, Igf2r, Grb10, Zim1, Peg3, Ndn, and Ipw were all unregulated; in particular, the expression of Zim1 was more than 10-fold higher, and the expression of Ipw was repressed in tetraploid fetuses. The methylation statuses of four DMRs H19, intergenic (IG), Igf2r, and Snrpn in tetraploid and triploid fetuses were similar to those in diploid fetuses. We also performed allele-specific RT-PCR sequencing to determine the alleles expressing the three imprinted genes Igf2, Gtl2, and Dlk1 in tetraploid fetuses. These three imprinted genes showed monoallelic expression in a parent-of-origin-specific manner. Expression of non-imprinted genes regulating neural cell development significantly decreased in tetraploid fetuses, which might have been associated with unregulated imprinted gene expression. This study provides the first detailed analysis of genomic imprinting in tetraploid fetuses, suggesting that imprinted gene expression is disrupted, but DNA methylation statuses of DMRs are stable following changes in ploidy in mammals

子宮内膜症性嚢胞の悪性転化におけるHO-1発現マクロファージの特徴

Malignant transformation of endometriosis is a rare and still poorly understood event, but is associated with the distortion of the pro-oxidant and anti-oxidant balance. The aim of the present study was to quantify the numbers of macrophages polarized as M1 or M2 phenotypes and the expression of heme oxygenase (HO)-1 in tissue sections from patients with benign ovarian endometrioma (OE) and its malignant transformation (endometriosis-associated ovarian cancer, EAOC). We performed a retrospective study at the Department of Gynecology, Nara Medical University hospital from December 2012 to March 2015. This study included 53 patients with OE (n = 33) and EAOC (n = 20), and we evaluated polarized functional status of macrophages by immunohistochemical staining of CD68, CD11c, CD163 and HO-1. The number of the M1 phenotype (CD11c+, p = 0.001) and the M2 phenotype (CD163+, p = 0.009) was significantly lower in EAOC patients than in OE patients. Analyzing the correlations between the studied markers, the expression of CD68, CD11c, and CD163 proteins significantly correlated with each other (p < 0.001). The number of M2 phenotypes expressing HO-1 was significantly decreased in the EAOC group, compared with the OE group (P < 0.001), demonstrating sustained downregulation of an antioxidant marker, HO-1, in EAOC. In conclusion, reduced number of M2 macrophages expressing HO-1 may have an important role in promoting malignant transformation of OE.博士（医学）・乙第1434号・令和元年9月27日Copyright © 2018. Published by Elsevier GmbH

Towards Better Ad Experience: Click Prediction Leveraging Sequential Networks Derived Specifically From User Search Behaviors

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Ehrlichia chaffeensis Infection of Sika Deer, Japan

To determine whether Ehrlichia chaffeensis exists in Japan, we used PCR to examine blood from sika deer in Nara, Japan. Of 117 deer, 36 (31%) were infected with E. chaffeensis. The E. chaffeensis 16S rRNA base and GroEL amino acid sequences from Japan were most closely related to those of E. chaffeensis Arkansas

Toll-like receptor 4 regulates gastric pit cell responses to Helicobacter pylori infection

Gastric pit cells express mitogen oxidase1 (Mox1) and essential components for the phogocyte NADPH oxidase (p67-, p47-, p40-, and p22-phoxes). Helicobacter pylori (Hp) lipopolysaccharide (LPS) is a potent up-regulator of the Mox1 oxidase. In this study, we examined the expression levels of several key members of the Toll-like receptor (TLR) family in primary cultures of guinea pig gastric pit cells. These cells expressed the TLR4 mRNA. Immunoblot analysis and immunofluorescence histochemistry with an anti-TLR4 antibody showed that gastric pit cells possessed significant amounts of TLR4 protein preferentially on the plasma membrane. In contrast, the cells did not express the TLR2 and TLR9 transcripts and did not contain detectable amounts of TLR2 protein. Neither peptidoglycan from Staphylococcus aureus nor Hp DNA with the CpG motif up-regulated Mox1 oxidase activity. Hp LPS activated nuclear factor-κB in association with the expression of cyclooxygenase II and tumor necrosis factor α transcripts. These findings suggest that TLR4 may play a crucial role in the initiation of inflammatory responses of gastric pit cells against Hp infection

急性期医療を受ける患者の願い

目的：患者の地域での暮らしの希望と療養上の目標を中心に構造化した看護のプロセス”Nursing Care for Patient Goals”（NCPG）に患者の視点から示唆を得るため急性期医療を受ける患者が看護師に知ってほしい情報と療養の目標に対して求める看護を明らかにした． 方法：１特定機能病院において入院中または入院予定の20歳以上の患者を対象として質問紙調査を２回（2017年，2018年）に実施した．調査内容は基本属性，調査Ⅰ（2017年）は先行研究を基に抽出した患者情報23項目について看護師に知って欲しいと思う程度，最も大事と思う項目とその選択理由，調査Ⅱ（2018年）は自分の療養の目標について求める看護であった．統計分析は記述統計，因子分析，t 検定を用いた．自由記述によるデータは質的記述的分析を行った． 結果：調査Ⅰの有効回答数は448名で，看護師に知ってほしい自分の情報として【第１因子：社会的役割と環境】【第２因子：病気の理解・受け入れと心理】【第３因子：身体的状態と生活の仕方】【第４因子：暮らしの希望と自己決定】が抽出された．65歳未満と比較して65歳以上の対象者は第１因子が高い傾向にあった．また第４因子を最も大事と思う項目の選択理由について， “希望・目標がなきゃ生きていけない” という表現が特徴として出された．調査Ⅱの有効回答数は416名で，多数の対象者が自分の目標を医療者と共有することが重要だと感じており，受けたい看護として，傍にいて寄り添う看護，治療・症状への専門的な看護，地域での暮らしの自立への看護が抽出された． 結論：患者の視点から看護師に知ってほしい情報として４つの因子と自分の目標を分ってほしいとする対象者の願いは，希望と目標を基盤としたNCPG の考え方と一致しており，目標達成のために受けたい看護の３つの視点が示唆された．Objective : This study aimed to identify information on care of patients receiving acute care for life in community from the patientsʼ perspective and to obtain suggestions for “Nursing Care for Patient Goals”（NCPG）. Method : The subjects were patients receiving acute care and aged 20 years and above. They were given self-administered questionnaires. Survey Ⅰ（2017）consisted of a questionnaire that was based on previously collected qualitative data and comprised 23 Likert-scale questions and free descriptive questions on the reasons for selecting the most important item. Survey Ⅱ（2018）consisted of questionnaire that was comprised of three Likert-scale questions on goals and a free descriptive question on care for the achievement of goals. Statistical analysis included descriptive statistics, factor analysis, and t-test. Data from free-text descriptions were analyzed using qualitative descriptive analysis. Results : Survey Ⅰ : data from 448 valid responses were subjected to factor analysis to determine the factor structure. The following factors were identified from the patientsʼ perspectives: 1） social role and environment, 2） understanding/acceptance and psychological state, 3） physical condition and life, and 4） hope and decision-making for life. In addition, a qualitative and inductive approach was employed to analyze participantsʼ descriptive responses about the reason for selecting the most important item. The characteristic description of why participants selected “hope and decision-making of life” was “I cannot live without hope.” Survey Ⅱ : data from 416 valid responses were analyzed. The majority of participants felt it was important to share their goals with their healthcare professionals. A qualitative and inductive approach was employed to analyze the participantsʼ descriptive responses to care for goals achievement. The care desired by participants was categorized as “being with”, “professional care”, and “self-care support”. Conclusion : The factors that patients wanted nurses to know were consistent with the components of “NCPG.” The care that patients desire to achieve their goals was clarified

Helicobacter pylori lipopolysaccharide from type I, but not type II strains, stimulates apoptosis of cultured gastric mucosal cells

The cag pathogenicity island (cag PAI) genes are a major determinant of virulence of Helicobacter pylori (Hp). Lipopolysaccharide (LPS) purified from the cag PAI-positive (type I) strains induced apoptosis of primary cultures of guinea pig gastric mucosal cells. Lipid A catalyzed this apoptosis. These cells expressed the Toll-like receptor 4 (TLR4) mRNA and its protein, and type I Hp LPS phosphorylated transforming growth factor β-activated kinase 1 (TAK1) and TAK1-binding protein 1 (TAB1) in association with up-regulation of the TLR4 expressions, suggesting that type I Hp LPS evoked distinct TLR4 signaling. In contrast, Hp LPS from type II strains with complete or partial deletion of the cag PAI genes did not phosphorylate TAK1 and TAB1 and failed to induce apoptosis. Accelerated apoptosis of gastric epithelial cells is one of the important events relevant to chronic, atrophic gastritis caused by Hp infection. The difference in proapoptotic action of LPS between the type I and II strains may support an important role of the cag PAI genes in the pathogenesis of gastric lesions caused by Hp infection

患者の希望を地域につなぐための患者状態とニーズ

目的：急性期医療を受ける患者の地域での生活を視野に入れた看護を展開するために患者の情報と看護の視点について明らかにすることを目的とした． 方法：特定機能病院に勤務する中堅以上の看護師３３名を対象に，地域での生活を視野に入れた患者の情報と看護の視点についてフォーカス・グループ・インタビューを行い分析した． 結果及び考察：急性期医療を受ける患者の地域での生活に必要な情報と看護の視点として，コアカテゴリー《地域での生活を可能にするニーズ》が抽出された．さらに地域での生活を可能にするための状態とニーズとして【身体・生理的な状態とニーズ】，【生活の自立と安全の状態とニーズ】，【病気の受け入れと心理的反応の状態とニーズ】，【社会的環境の状態とニーズ】，【医療・療養への自己決定の状態とニーズ】の５つのカテゴリーに分類された．これらより，患者の暮らしの希望，療養の目標，５つの視点の状態からニーズを導き看護を展開する看護の過程として，“Nursing Care for Patient Goals（” NCPG）を構造化した． 結論：地域包括ケアシステムの中において急性期医療を受ける患者の情報と看護の視点として地域での生活を可能にするための５つの状態とそのニーズが重視されていた．Objective : The aim of this study was to identify information and care perspectives of nurses for patients leaving an acute care hospital for life in the community, and to consider appropriate nursing care in the community-based integrated care system. Method : Focus group interviews were conducted with 33 nurses working in an acute care hospital. The data were analyzed using qualitative inductive analysis. Results & Discussion : The core category “Needs to enable patients to live in the community” was extracted as the information and care perspective necessary for patients receiving acute care to leave hospital for life in the community. The information and care perspectives were classified into five conditions : physical/physiological condition and needs, life independence and safety status and needs, acceptance of/emotions about illness and needs, social environment and needs, and decision-making and needs. “Nursing Care for Patient Goals” was structured as a nursing process. Conclusion : We identified five conditions and their needs that would enable patients receiving acute care to leave hospital for life in the community

Activation of ERK/IER3/PP2A-B56γ-positive feedback loop in lung adenocarcinoma by allelic deletion of B56γ gene

In order to investigate the involvement of the IER3/ PP2A-B56γ/ERK-positive feedback loop, which leads to sustained phosphorylation/activation of ERK in carcinogenesis, we immunohistochemically examined the expression of IER3 and phosphorylated ERK in lung tumor tissues. IER3 was overexpressed in all cases of adenocarcinomas examined, but was not overexpressed in squamous cell carcinomas. Phosphorylated ERK (pERK) was also overexpressed in almost all adenocarcinomas. EGFR and RAS, whose gene product is located upstream of ERK, were sequenced. Activating mutation of EGFR, which is a possible cause of overexpression of IER3 and pERK, was found only in 5 adenocarcinomas (42%). No mutation of RAS was found. We further examined the sequences of all exons of B56γ gene (PPP2R5C) and IER3, but no mutation was found. Using a single nucleotide insertion in intron 1 of PPP2R5C, which was found in the process of sequencing, allelic deletion of PPP2R5C was examined. Eight cases were informative (67%), and the deletion was found in 4 of them (50%). Three cases having deletion of PPP2R5C did not have EGFR mutation. Finally, PPP2R5C deletion or EGFR mutation that could be responsible for IER3/pERK overexpression was found in at least 8 cases (67% or more). This is the first report of a high incidence of deletion of PPP2R5C in human carcinomas.Embargo Period 12 month

1-year tolvaptan efficacy in ADPKD

Autosomal dominant polycystic kidney disease (ADPKD) develops into end-stage kidney disease by 65 years of age in an estimated 45%-70% of patients. Recent trials revealed that tolvaptan inhibits disease progression both in early-stage or late-stage ADPKD ; however, stratified analysis showed a difference of favorable factors correlated with tolvaptan efficacy between early-stage and late-stage ADPKD. Thus, we examined the efficacy of tolvaptan in ADPKD with a wide range of estimated glomerular filtration rates (eGFR). We enrolled 24 patients with eGFR 35.3 (28.0-65.5) ml / min / 1.73m2 and evaluated treatment effect as ΔΔeGFR (ml / min / 1.73m2 / year) or ΔΔtotal kidney volume (TKV) (% / year) that was calculated as post-treatment annual change - pre-treatment annual change. Pre ΔeGFR was significantly low in eGFR responders, defined as ΔΔeGFR > 0 ml / min / 1.73m2 / year. In eGFR responders, pre ΔeGFR, post ΔeGFR, eGFR, TKV, and proteinuria were significantly correlated with ΔΔeGFR. In TKV responders defined as ΔΔTKV > 5 % / year, we identified hypertension history, proteinuria, TKV, and post ΔTKV as significantly correlated factors with ΔΔTKV. In conclusion, pre ΔeGFR may be a predictive factor of therapeutic efficacy on kidney function. Tolvaptan may have greater efficacy in early-stage ADPKD with rapid GFR decline or with well-controlled blood pressure