Rheumatoid arthritis (RA) is an inflammatory disease with joint dysfunction following cartilage degradation. The level of lysophosphatidic acid (LPA) has been reported to be augmented in human synovial fluid from patients with RA. However, it remains to be elucidated whether LPA participates in cartilage destruction. In the present study, we have demonstrated that the production of promatrix metalloproteinases (proMMPs)-1 and -3 was augmented along with an increase of extracellular signal-regulated kinase (ERK)1/2 phosphorylation through LPA receptor 1 (

) in human synovial fibroblasts. These results suggest that LPA transcriptionally increases MMP production by the activation of an /ERK1/2 signal pathway in human synovial fibroblasts. Thus, LPA is likely to be a pathological candidate for cartilage degradation in RA.

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Lysophosphatidic acid (LPA) is a bioactive mediator and induces several biological effects, including cell proliferation, migration, morphogenesis and differentiation. LPA interacts with at least six G protein-coupled receptors (GPCRs), including LPA receptor-1 (LPA₁), LPA₂, LPA₃, LPA₄, LPA₅ and LPA₆. These receptors show different biological functions through the binding of LPA, depending on the type of cells. In human malignancies, a high level of LPA production was found in plasma and ascites in ovarian cancer cases. Moreover, aberrant expression levels of LPA receptor genes were detected in some cancer cells. Therefore, it is suggested that LPA receptors may be involved in the pathogenesis of tumor cells as well as LPA per se. Recently, we have reported that alterations of LPA receptor genes also occur in rodent tumors. In this review, we summarize the recent evidence in the investigations of LPA receptor alterations in rodent tumors by experimental models.

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【目的】リゾフォスファチジン酸(LPA)は細胞膜受容体を介して多様な生理機能に関与する脂質メディエーターであり，主に酵素オートタキシン(ATX)により産生される。妊娠高血圧症候群(PIH)では，正常妊娠と比較して胎盤におけるATX産生が有意に低下することを我々は過去に報告した。本研究ではATX産生低下の意義を調べるため，LPA受容体の一つである3に着目した。3は，その欠損マウスにおいて着床遅延が生じることが報告されており，生殖分野への関与が示唆されている。我々は，3の胎盤における発現パターンを検討した。また絨毛細胞由来株に3を強制発現させた上でアゴニストによる刺激を行い，血管新生関連因子の発現を検討した。【方法】本研究は当院研究倫理委員会の承認の下，患者の同意を得て行った。免疫染色法によりヒト胎盤におけるATX，3の発現分布を確認した。絨毛細胞由来株HTR-8/SVneoに3を遺伝子導入し，強制発現されることを確認した。その上で3特異的アゴニストT13による刺激を行い，血管新生関連因子の発現を定量的PCR法を用いて同定した。【成績】ATXはすべての絨毛細胞に発現していたが，3の発現は分化したsyncytiotrophoblast，extra-villous trophoblastに限局していた。3を強制発現したHTR-8/SVneoをT13にて刺激すると，血管新生関連因子の中でCOX2，VEGF，IL8の発現が誘導された。【結論】ATX-LPA-3系は，分化した絨毛細胞における血管新生関連因子の発現を調節していることが示唆された。このことから，ATXの産生低下は胎盤局所の血管新生関連因子の発現低下を通じて妊娠初期の胎盤形成不全をもたらし，PIHの発症につながる可能性が示されたと考える。

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We examined whether the CL is a site for lysophosphatidic acid (LPA) synthesis and/or a target for LPA action in the bovine reproductive tract. LPA concentrations in the CL tissue increased towards the end of the cycle and were stable during early pregnancy. No changes in the expression of LPA receptors (LPARs) occurred during the estrous cycle. The expressions of 2 and 4 on days 17–19 of pregnancy were higher than those on the respective days of the estrous cycle and higher than those on days 8–10 of pregnancy. LPA stimulated P4 synthesis via 3βHSD stimulation but did not modulate the interferon–tau (IFNτ) influence on P4 synthesis in steroidogenic cells. Moreover, we found LPA-dependent stimulation of IFNτ action on 2,5’-oligoadenylate synthase (OAS1) and ubiquitin-like IFN-stimulated gene 15-kDa protein (ISG15) expression. The present study demonstrated that the CL might be a site of LPA synthesis and target of LPA action in the bovine reproductive tract. We postulate that during the estrous cycle and early pregnancy, LPA exerts autocrine and paracrine effects on the CL mainly via and

LPAR

4. The stimulatory effect of LPA on P4 synthesis via 3βHSD stimulation and LPA-dependent stimulation of IFNτ action on OAS1 and ISG15 expression suggest that LPA is an additional auxiliary luteosupportive factor in steroidogenic cells.

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The tumor promoting agent 12-O-tetradecanoylphorbol-13-acetate (TPA) stimulates cell migration of several tumor cells. Recently, we reported that loss of lysophosphatidic acid (LPA) receptor-3 (LPA₃) enhanced cell migration of murine lung tumor LL/2 cells. In the present study, we investigated whether LPA₃ is involved in cell migration of mouse lung tumor cells stimulated by TPA. Exogenous LPA₃ gene ()-expressing (LL/2-a3) cells and LL/2-AB cells as a vector control generated from LL/2 cells were used. In a cell migration assay, TPA treatment significantly stimulated cell migration of LL/2-AB and LL/2-a3 cells, while the cell migration abilities of LL/2-a3 were markedly lower than those of LL/2-AB cells. Using quantitative real-time reverse transcription (RT)-polymerase chain reaction (PCR) analysis, no effect of TPA treatment on the expression levels of LPA₁, LPA₂ and LPA₃ genes was detected in either type of cells. These results suggest that the LPA₃ may not be involved in the enhanced migration ability by TPA in mouse lung tumor cells.

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Lysophospholipids are phospholipids with only one fatty acid. During the past two decades, it has become apparent that lysophospholipids are not merely degradation products but have various physiological and pathological functions in vivo via G protein-coupled receptor (GPCR)-type receptors. These include lysophosphatidic acid (LPA), sphingosine 1-phosphate (S1P), lysophosphatidylinositol/lysophosphatidylglucose (LPI/LPtdGlc), and lysophosphatidylserine (LysoPS). This review focuses on identifying the functions of the receptors, enzymes, transporters, and carrier proteins required for these four lysophospholipids to function as lipid mediators. We also note that many of advances in this field have been made by Japanese pharmaceutical scientists.

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Lysophosphatidic acid (LPA) acts as a simple phospholipid that interacts with G protein-coupled transmembrane LPA receptors. Recently, it has been reported that each LPA receptor plays different biological roles in acquisition of the malignant property of tumor cells. In this study, to assess the involvement of LPA receptor-3 (LPA₃) in cell survival after treatment with anticancer drugs, we generated -expressing FM3A-a3A9 cells from mouse mammary tumor FM3A cells and examined the cell survival rate after treatment with anticancer drugs compared with -unexpressing cells. Cells were treated with 0.005 to 10 μM of cisplatin (CDDP) or doxorubicin (DOX) for 3 days. For the CDDP and DOX treatments, the cell survival rate of FM3A-a3A9 cells was significantly higher than that of -unexpressing cells. The expression level of the Mdr1a gene in FM3A-a3A9 cells was higher than that of -unexpressing cells, whereas no significant difference in multidrug resistance 1b (Mdr1b) and glutathione S-transferase mu1 (Gstm1) expressions was found. These results suggest that LPA₃ may enhance the cell survival rate after treatment with anticancer drugs in mouse mammary tumor cells, correlating with increased expression of the Mdr1 gene.

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Skin barrier is important for the moisture maintenance and function of our skin. One of the most important gene that constitutes our skin barrier is filaggrin (FLG). Deficits in FLG expression results in skin barrier dysfunction and causes atopic dermatitis (AD). To elucidate molecular mechanism maintaining skin barrier, we sought to identify factors that could enhance FLG gene expression in human keratinocytes, and evaluate the effect of such factors under physiological condition.

　We first identified G-protein coupled receptors (GPCRs) highly expressed in normal human epidermal keratinocytes (NHEKs) using GPCR array and next examined effects of agonists of these GPCRs on FLG expression by qRT-PCR and Western blot. We then validated GPCRs mediating the above effects on FLG gene expression, using selective agonists/antagonists and small interfering RNA (siRNA). We further conducted comprehensive analysis of gene expression induced by their activation, and identified signaling pathway leading to this induction by using pharmacological inhibitors. Finally, we tested the potential of this pathway for promoting skin barrier function under physiological condition by applying an agonist of the identified GPCRs to the 3D culture of human skin and experimental mice.

　In summary, we identified that lysophosphatidic acid (LPA) could induce keratinocyte differentiation in NHEKs. LPA signals through

1 and 5 and then activate Rho-ROCK-SRF signaling pathway indispensable for expression of variety of skin barrier genes including FLG. Moreover, we demonstrated that LPA treatment significantly promoted skin barrier function in vivo. Therefore, 1 and 5 signaling may serve as a new therapeutic target for diseases associated with skin barrier dysfunction such as AD.

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本論文ではプロジェクト開始時における,迅速,簡易,客観的な見積り要求に対応し,かつ,カスタマイズ可能な概算工数見積りモデルを提案する.従来の係数モデルはパラメータおよび係数値等が不変であるが,当モデルはカスタマイズ可能なモデルとして新規性がある.また,柔軟な対応能力の高い手法として有用性がある.常に進化するIT業界において,最新の状況を反映するためモデルは見直しをかけられるべきであり,またプロジェクト状況に応じた変更がなされるべきであるため,パラメータおよび係数値の見直し方法を提示する.本手法を以下に示す.(1)プロジェクト分析を行い,標準WBSを作成する(2)規模と難易度要因のパラメータを定義する(3)難易度係数をWBSより求め,難易度要因を難易度係数を用いて規模に変換する(4)規模を工数に変換するための工数係数を求める.この工数係数のカスタマイズ方法を提示する.(5)工数係数により,規模と規模で表された難易度をプロジェクトの全体工数に変換する.本手法を実際のプロジェクトの概算工数見積りに適用した.算出工数と実工数の差は10%以内であり,有効な算出手法であることが実証された.

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Though lysophosphatidic acid (LPA) shows a variety of regulatory roles in reproduction, its action mechanisms in the gestational organs are still largely unknown. We here characterized cellular distribution of its six kinds of specific receptors (LPA1-6) in rat uteri by immunohistochemistry and quantitatively analyzed changes in

1-6 mRNAs expression throughout pregnancy. Among LPA1-6, evident expression of LPA3, LPA4, and LPA6 was immunologically detected and less expression of immunoreactive LPA1 and LPA2 was also found. Luminal and glandular epithelial cells, stromal cells, and myometrial cells are sites of positive immunoreactions, and they are all likely to express three or more subtypes. All of

Lpar

1-6 mRNAs were expressed, and their alterations were variable depending on subtypes and gestational age. The present information suggests that diverse actions of LPA in the uterus involve varied expression of LPA receptors dependent on tissue/cell types, receptor subtype(s), and organ reproductive states and helps to understand uterine biology of LPA.

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【目的】われわれは，エストロゲン誘発性乳腺腫瘍の形成に関連するメタボローム変化として血清中リゾホスファチジルコリン（LPC）が有意に上昇することを明らかにしている．本研究では，LPCが乳腺組織に及ぼす影響を明らかにするために，LPCおよびその活性代謝物リゾホスファチジン酸（LPA）のヒト乳がんMDA-MB-231細胞に対する増殖促進作用について検討した．

【方法】被験物質：LPC (18:1)，LPA (18:1)およびラット血清（乳腺腫瘍を形成したラット由来）．細胞増殖試験：MDA-MB-231に対して被験物質を2日間処理したのち，細胞数を測定．細胞数のカウントには，hoechst33342で核染色後，ハイコンテントイメージングシステムを使用．遺伝子発現解析：MDA-MB-231細胞におけるオートタキシン（ATX；LPCをLPAへ変換する酵素）およびLPA受容体（

）のmRNA発現をRT-PCRにより確認．

【結果および考察】ヒト乳がんMDA-MB-231細胞はATXおよび

，遺伝子を発現していることが，遺伝子発現解析の結果から明らかとなった．細胞増殖試験において，LPC (18:1)およびLPA (18:1)，ラット血清は用量依存的に細胞数の増加を引き起こした．続いて，ATXおよび1/2の関与を検証するために，S32826（ATX阻害剤）およびKi16425（1/3阻害剤）のMDA-MB-231細胞増殖に対する抑制効果を評価した．その結果，Ki16425はLPC (18:1)およびLPA (18:1)，ラット血清による細胞増殖促進作用をコントロールレベルまで抑制した．したがって，この細胞増殖は1による細胞内シグナルの活性化を介していることが示唆される．一方，S32826はLPC (18:1)およびラット血清による作用を抑制したが，LPA (18:1)処理に対しては何の影響も及ぼさなかった．以上の結果から，ラット血清中に含まれるLPCはATXによってLPAへと変換されたのち，1を介して細胞増殖を促進していることが示唆される．

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Public key authenticated encryption with keyword search (PAEKS) has been proposed, where a sender's secret key is required for encryption, and a trapdoor is associated with not only a keyword but also the sender. This setting allows us to prevent information leakage of keyword from trapdoors. Liu et al. (ASIACCS 2022) proposed a generic construction of PAEKS based on word-independent smooth projective hash functions (SPHFs) and PEKS. In this paper, we propose a new generic construction of PAEKS, which is more efficient than Liu et al.'s in the sense that we only use one SPHF, but Liu et al. used two SPHFs. In addition, for consistency we considered a security model that is stronger than Liu et al.'s. Briefly, Liu et al. considered only keywords even though a trapdoor is associated with not only a keyword but also a sender. Thus, a trapdoor associated with a sender should not work against ciphertexts generated by the secret key of another sender, even if the same keyword is associated. That is, in the previous definitions, there is room for a ciphertext to be searchable even though the sender was not specified when the trapdoor is generated, that violates the authenticity of PAKES. Our consistency definition considers a multi-sender setting and captures this case. In addition, for indistinguishability against chosen keyword attack (IND-CKA) and indistinguishability against inside keyword guessing attack (IND-IKGA), we use a stronger security model defined by Qin et al. (ProvSec 2021), where an adversary is allowed to query challenge keywords to the encryption and trapdoor oracles. We also highlight several issues associated with the Liu et al. construction in terms of hash functions, e.g., their construction does not satisfy the consistency that they claimed to hold.

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本論文は,ハイダ語スキドゲイト方言の音韻論においてこれまで十分解明されていなかったピッチ(声調;高,中,低の3つがある)について論じたものである.Hori(1994b)において,それぞれのピッチの現われが音節構造と関係があり,予測可能であることから,ピッチは音声レベルに属するものと捉え,ピッチの音声的実現を導く5つのピッチ付与規則を設定した.本論文では,Hori(1994b)が十分考慮にいれなかった形態レベルに関する情報がピッチ付与に関わっていることを指摘し,それらの規則の条件を若干修正する必要があると考える.また,そこで設定した5つの規則のうち,2つの規則は,「低ピッチ連結規則」の一部と見なすことができ,超分節音的な諸現象を説明するためには,更に新たな規則を1つ設定しなくてはならない.結局,計4つのピッチ付与規則によって,同言語のピッチの音声的実現を導くことができるというのが本論文の結論である.

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【背景】ウシにおいて配偶子および初期胚の輸送に必須である卵管平滑筋の収縮および弛緩運動は，それぞれPGF2αおよびPGE2により制御される。LPAは生体内で多様な生理作用を有するリン脂質であり，ウシ子宮内膜細胞においてPG分泌を促進することが示されている。本研究では，卵管平滑筋運動の制御機構を明らかにする目的で，ウシ卵管におけるLPA関連遺伝子の発現，卵管上皮および間質細胞のPG分泌に及ぼすLPAの影響について検討した。【方法】1) ウシ卵管を排卵後2–5日，10–12日，16–18日，20–21日に分類して採取し，組織中LPA濃度を測定した。2) 卵管組織におけるLPA受容体 (

LPAR

1-6) およびLPA産生酵素 (ATX，PLA1) mRNA発現を検討した。3) 卵管膨大部および峡部からそれぞれ単離した上皮ならびに間質細胞における

LPAR

1-6およびLPA産生酵素のmRNA発現を検討した。更に，卵管培養上皮および間質細胞にLPA (0.1，1，10 μM) を添加し，4) 4時間後のPG合成酵素 (COX-1，COX-2) mRNA発現，5) 24時間後の培養上清中におけるPGF2αおよびPGE2濃度を測定した。【結果】1) 卵管組織中のLPA濃度は膨大部では排卵後2–5および10–12日，峡部では排卵後2–5日に高かった。2)

LPAR

1-6およびATX，PLA1 mRNA発現は卵管膨大部ならびに峡部の両方に認められた。また，

LPAR

1-3 mRNA発現は峡部より膨大部において，

LPAR

4-6 mRNA発現は膨大部より峡部において高かった。3)

LPAR

1-6，ATX，PLA1 mRNA発現は卵管の部位にかかわらず，上皮ならびに間質細胞に認められた。4) LPAは峡部間質細胞においてのみCOX-2 mRNA発現を刺激した。5) COX-2の結果と同様に，LPAは峡部の間質細胞でのみPG分泌を刺激した。以上より，LPAは卵管峡部間質細胞のCOX-2合成を促進することにより，PG分泌を刺激し，平滑筋運動を制御する重要な因子であると考えられる。また，LPAは峡部において高い発現がみられたLPA受容体4-6を介して，間質細胞のPG分泌を刺激している可能性が示された。

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We investigated mechanism of the dissolution of HBPE-NO₂ films to the developer by analysis of the developer after development. A HBPE-NO₂ film containing BADNQ2 was irradiated with 500 mJ/cm² of exposure dose and developed with 25wt% TMAHaq. The developer was then neutralized by HClaq. and the solution was evaporated and dried at 90 °C in vacuo. ¹H-NMR spectrum of the resulting product after development is shown in Fig. 3. The 1H-NMR spectrum shows signals assigned to the monomers, terephthalic acid derived from TPC and THPE, and 4-nitrobenzoic acid derived from the end groups. However, no signal that can be assigned to the original HBPE-NO₂ protons was observed. This result suggests that RDP of HBPE-NO₂ is not based on solubilization of the polymer by the reaction at the end groups but on decomposition of the polymer induced by nucleophilic attack of the OH^- to the ester linkages to give low-molecular-weight products.

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Most of photoautotrophic microalgal culture is limited by light because light is easily attenuated in algal suspensions. In order to predict the microalgal growth, the spatial light distribution within photobioreactors and the light-dependent algal activity should be understood and mathematically modeled. In general, CO₂-enriched air is bubbled into photobioreactors and the bubbles, as well as algal cells, are able to scatter light. This makes it difficult to obtain high-quality data sets of spatial light distribution and eventually of light-dependent microalgal growth.
In this study, an airlift rectangular photobioreactor was designed and used for kinetic analysis of light-limited growth of green microalga Chlorella vulgaris. Since light was illuminated only into the down-comer where bubbles did not exist, the effect of light scattering by air bubbles was avoided and the growth curves at various photon flux densities could be obtained under well-defined light conditions. It was found that the effect of light on the specific growth rate was successfully explained by the models based on the local photon flux density (LPFD) and local photon absorption rate () hypotheses rather than average photon flux density (APFD) and average photon absorption rate (APAR) hypotheses. Using the developed models, the algal growth was satisfactorily simulated at various light-related conditions such as incident light intensity, light path length (size of photobioreactor), surface to volume ratio, and algal concentration. Consequently, the presented models could be used for predicting the light-limited algal growth, if necessary, when further expanded to general cases like bubble-existing system.

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Separation logic is an extension of Hoare logic to verify imperative programs with pointers and mutable data-structures. Although there exist several implementations of verifiers for separation logic, none of them has actually been itself verified. In this paper, we present a verifier for a fragment of separation logic that is verified inside the Coq proof assistant. This verifier is implemented as a Coq tactic by reflection to verify separation logic triples. Thanks to the extraction facility to OCaml, we can also derive a certified, stand-alone and efficient verifier for separation logic.

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ITインフラ構築プロジェクトでは,アプリケーション・コードのような工数算出のベースがないため,モデル化された工数算出手法がない.工数見積りの誤りはプロジェクトの失敗の原因となるため客観的な算出手法が求められる,本論文ではプロジェクト開始時における概算工数見積りモデルを提案する.本手法を以下に示す.(1)プロジェクト分析を行い,工数算出の要素としての規模と難易度要因を明確にする.(2)難易度要因を難易度係数を用いて規模で表現する.(3)規模と規模で表された難易度からプロジェクトの全体工数を求める.工数はプロジェクト実績の分析により考案したプロジェクトの工数算出式を用いる.本手法を実際のプロジェクトの工数見積りに適用し,有効な工数算出モデルであることが実証できた.

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Purpose : This study examined differences in the viscoelasticity of the periodontium between Japanese and Chinese subjects and the influence of their living environment on the periodontium.
Materials and Methods : We selected 93 and 113 upper central incisors, 100 and 100 upper canines, and 104 and 98 upper first molars of the Japanese and Chinese subjects, respectively, for examination in this study. The viscoelasticity of the periodontium was measured using an automatic diagnostic system that measured tooth mobility. The evaluated parameters were viscosity c₁, c₂ and elasticity k. An analysis of variance was used to compare the upper central incisors, canines, and first molars in the Japanese and Chinese subjects. An unpaired t-test was used for comparison between sexes and between Japanese and Chinese.
Results : The viscoelastic parameters showed significant differences among central incisors, canines, and first molars in both the Japanese and Chinese. Almost all periodontal viscoelastic parameters were significantly different between Japanese males and females, while only a few such parameters showed significant gender differences in the Chinese. All parameters except c₁ of the central incisors in men and all parameters in women showed significant differences between the Japanese and Chinese subjects.
Conclusion : There were differences in the viscoelastic properties of the periodontium between Japanese and Chinese subjects.

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We firstly found that lysophosphatidic acid receptor 1 (LPA1) signaling initiates nerve injury-induced neuropathic pain (NP) in mice (Nature Medicine, 2004) and LPA1 and LPA3 signaling are also involved in the development and maintenance of various mouse models of chronic pain, such as peripheral neuropathic paclitaxel-induced pain (Mol Pain, 2014) and central emotional fibromyalgia (Neurobiol of Pain, 2017). Here we studied to see whether LPA1 and LPA3 signaling are also involved in the central NP, central post-stroke pain (CPSP), which is one of the most intractable chronic pain syndromes in cerebral ischemia, though useful treatment has not been established. The CPSP model in our study was developed by the late stage of tPA treatment 6 h after photochemically induced thrombosis (PIT) of left middle cerebral artery (MCA). Significant abnormal pain behaviors in the thermal and mechanical test were found on both sides for more than 2 weeks, and they were abolished in

1 and 3 KO. In addition, the repeated treatments with Ki-16425, an LPA1/3 receptor antagonist also abolished the abnormal pain behaviors. When LPA levels were measured by LC-MS/MS, tPA-dependent increase in LPA levels were found in some brain regions. We will discuss why tPA-accelerated bilateral CPSP occurs in the PIT stroke model on the left side.

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