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The sonic hedgehog (Shh) pathway is activated in ∼30% of human medulloblastoma resulting in increased expression of downstream target genes. In about half of these cases, this has been shown to be a consequence of mutations in regulatory genes within the pathway, including Ptc1, Smo, and Sufu. However, for some tumors, no mutations have been detected in known pathway genes. This suggests that either mutations in other genes promote tumorigenesis or that epigenetic alterations increase pathway activity in these tumors. Here, we report that 3% to 4% of mice lacking either one or both functional copies of Cxcr6 develop medulloblastoma. Although CXCR6 is not known to be involved in Shh signaling, tumors derived from Cxcr6 mutant mice expressed Shh pathway target genes including Gli1, Gli2, Ptc2, and Sfrp1, indicating elevated pathway activity. Interestingly, the level of Ptc1 expression was decreased in tumor cells although two normal copies of Ptc1 were retained. This implies that reduced CXCR6 function leads to suppression of Ptc1 thereby increasing Smoothened function and promoting tumorigenesis. We used a direct transplant model to test the sensitivity of medulloblastoma arising in Cxcr6 mutant mice to a small-molecule inhibitor of Smoothened (HhAntag). We found that transplanted tumors were dramatically inhibited in mice treated for only 4 days with HhAntag. These findings suggest that HhAntag may be effective against tumors lacking mutations in known Shh pathway genes. [Cancer Res 2007;67(8):3871–7]

Supplementary Figure 1 and Tables 1-2 from Medulloblastomas Derived from Cxcr6 Mutant Mice Respond to Treatment with a Smoothened Inhibitor

Background: Human resource management may be difficult for community pharmacists because of recent drastic changes in work volume and content. In Japan, few studies have been conducted on job satisfaction, well-being, and quality of work life (QWL) among community pharmacists. Focusing on QWL, a more comprehensive concept than job satisfaction, this study aimed to develop a multi-trait-based QWL scale for Japanese community pharmacists (QWLS for JCP) and test its reliability and validity. Methods:A questionnaire survey was conducted among 2027 pharmacists working in pharmacies, with the cooperation of 20 corporations running pharmacies. The collected data were subjected to principal component factor analysis with Promax rotation using SPSS28. Results: The following five significant components were identified based on factor analysis using data from 1966 pharmacists. These form the basis of the QWLS for JCP: “Influence of work on mind and body,” “Relationships with colleagues,” “Relationship with the boss,” “Meaning of existence in the workplace,” and “Pride in work.” Cronbach’s alpha, which expresses reliability, was between 0.585 and 0.854 for these subscales. Conclusions: The QWLS for JCP significantly explained the concept of QWL, which means that its validity was sufficient.

Deoxyhypusine synthase (DHPS) is the primary enzyme responsible for the hypusine modification and, thereby, activation of the eukaryotic translation initiation factor 5A (eIF5A), which is key in regulating the protein translation processes associated with tumor proliferation. Although DHPS inhibitors could be a promising therapeutic option for treating cancer, only a few studies reported druglike compounds with this inhibition property. Thus, in this work, we designed and synthesized a new chemical series possessing fused ring scaffolds designed from high-throughput screening hit compounds, discovering a 5,6-dihydrothieno[2,3-c]pyridine derivative (26d) with potent inhibitory activity; furthermore, the X-ray crystallographic analysis of the DHPS complex with 26d demonstrated a distinct allosteric binding mode compared to a previously reported inhibitor. These findings could be significantly useful in the functional analysis of conformational changes in DHPS as well as the structure-based design of allosteric inhibitors.

The establishment and implementation of a healthy lifestyle is fundamental to public health and is an important issue for working-aged people, as it affects not only them but also the future generations. However, due to the COVID-19 pandemic and associated behavioural restrictions, lifestyles have altered, and, in certain environments, significantly worsened. In the present study, we conducted a project to improve the intestinal environment by focussing on the dietary habits of participants, utilising the living laboratory as a social technology to explore how to adapt to this drastic environmental change. We held eight workshops for voluntary participants and implemented a self-monitoring process of recording dietary behaviours (n = 78) and testing the intestinal environment (n = 14). Through this initiative, we developed a personalised wellness enhancement programme based on collaboration with multiple stakeholders and a framework for using personal data for research and practical purposes. These results provide an approach for promoting voluntary participation and behavioural changes among people, especially under the COVID-19 pandemic, as well as a practical basis for the government, academia, and industry to intervene effectively in raising people’s awareness of health and wellness.

Scholarly attention to pharmaceutical companies' ability to sustain research and development (R&D) productivity has increased as they increasingly handle business challenges. Furthermore, the deterioration of R&D productivity has long been considered a major cause of mergers and acquisitions (M&As). This study attempts to investigate quantitatively the possible causes of the deterioration and the relationship between the deterioration and M&As by examining the Japanese pharmaceutical industry. Japan from 1980 to 1997 is an ideal case because of the availability of official data, but more importantly the significant changes in its business environment at the time. Using the Malmquist Index and data envelopment analysis, we measured the deterioration of R&D productivity from 1980 to 1997 based on a sample of 15 Japanese companies. Two lessons can be learned from Japan's case. First, to sustain R&D productivity over the long term, companies should use licensing activities and focus on the dominant therapeutic franchises. Second, if a company fails significantly to catch up with the benchmark, it is likely to pursue an M&A or seek an alternative way to improve R&D productivity. These findings appear similar to the current situation of the global pharmaceutical industry, although Japan pursued more licensing activities than M&A to improve R&D productivity.

製薬業界は，大型製品の特許満了，研究開発の遅れ，新規上市品目数の減少，後発医薬品の市場浸透，薬価制度の変更と，近年，急速に事業環境が大きく変化しており，新たなビジネスモデルの確立が必要となってきている。これらの問題に対しての打開策として，海外大手製薬企業は，他社との経営統合，新興諸国市場への進出，後発医薬品事業の展開など，新しいビジネスモデルの構築を行っており，一方，自社開発製品の海外展開により成長してきた国内大手製薬企業は，海外企業の買収等を行うことで，打開策を模索している。国内では，海外に比べ遅れていた後発医薬品の使用促進が国の主導のもと行われており，国内事業に特化している新薬開発型製薬企業にとっては，製品の特許満了後の後発医薬品へのシフトによる減収リスクが大きくなってきており，収益を長期収載品に依存するビジネスモデルの変革を余儀なくされている。このように，事業環境の変化に対応すべく，近年，製薬企業が取っている動きの大きなポイントとなるものは，1）医薬品兼業企業による経営統合，2）新興諸国への取組み強化，3）後発医薬品の取組み強化，4）企業買収による海外展開に分けられる。今後のジネスモデルの確立と成功が待たれる。

Background: Eukaryotic initiation factor 4A3 (eIF4A3) is a member of the Asp-Glu-Ala-Asp (DEAD) box RNA helicase family. There are three subtypes of eIF4A, eIF4A1, 2, and 3. eIF4A1 and eIF4A2 are required for translation initiation as their name suggest, however eIF4A3 is functionally distinct and one of the core components of the exon junction complex (EJC). The EJC is known to be involved in a variety of RNA metabolic processes typified by nonsense-mediated RNA decay (NMD), which is the surveillance mechanism that recognizes mRNAs containing premature termination codons to prevent the accumulation of truncated proteins. It is known that eIF4A3 is an ATP-dependent RNA clamp that can serve as a nucleation center to recruit other EJC components, and siRNA-mediated knockdown of eIF4A3 leads to a defect in NMD. In order to investigate the functions of eIF4A3 further and evaluate the therapeutic potential, we conducted a search for molecular probes of eIF4A3. Methods: Using an RNA dependent ATPase assay as a guide, intensive structure activity relationship (SAR) study and chemical optimization of high-throughput screening hit were conducted. Thereafter, NMD inhibitory activity, selectivity, helicase inhibitory activity, and physicochemical properties of optimized compounds were confirmed. Additionally, direct binding of inhibitors to eIF4A3 and their binding mode were analyzed using biophysical methods. Results: Optimized compounds showed high selectivity for eIF4A3 and exhibited significant NMD inhibitory activity in HEK293T cells. The drastic difference in eIF4A3 inhibitory activity between eutomers and distomer revealed the importance of stereochemistry at the 3-position of the piperazine ring for eIF4A3 inhibition. The lack of significant NMD inhibition by distomers suggests that NMD inhibition of eutomers is mediated by eIF4A3 inhibition. A surface plasmon resonance (SPR) biosensing assay and hydrogen/deuterium exchange mass spectrometry (HDX MS) provided strong evidence for the direct binding of the inhibitors to eIF4A3 and implications for their binding site. Conclusion: We discovered the first selective eIF4A3 inhibitors exhibiting cellular NMD inhibitory activity. These novel inhibitors not only represent novel molecular probes for investigation of RNA biology but also could be promising lead compounds for drug discovery research. References: [1] Ito, M.; Tanaka, T.; Cary, D. R.; Iwatani-Y, M.; Kamada, Y.; Kawamoto, T.; Aparicio, S.; Nakanishi, A.; Imaeda, Y. Discovery of novel 1,4-diacylpiperazines as selective and cell-active eIF4A3 inhibitors. J. Med. Chem. 2017, 60, 3335-3351. [2] Iwatani-Y, M.; Ito, M.; Ishibashi, Y.; Oki, H.; Tanaka, T.; Morishita, D.; Ito, T.; Kimura, H.; Imaeda, Y.; Aparicio, S.; Nakanishi, A.; Kawamoto, T., Discovery and characterization of a eukaryotic initiation factor 4A-3-selective inhibitor that suppresses nonsense-mediated mRNA decay. ACS Chem. Biol. 2017, 12, 1760-1768. Citation Format: Masahiro Ito, Misa Iwatani-Yoshihara, Toshio Tanaka, Douglas R. Cary, Yusuke Kamada, Yoshihiro Ishibashi, Hideyuki Oki, Satoshi Sogabe, Shoichi Nakao, Daisuke Morishita, Takashi Ito, Hiromichi Kimura, Tomohiro Kawamoto, Samuel Aparicio, Atsushi Nakanishi, Yasuhiro Imaeda. Discovery of the first selective and cell-active eIF4A3 inhibitors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2802.

SummaryThe Hedgehog (Hh) pathway plays critical roles in normal development and in tumorigenesis. We generated Gli-luciferase transgenic mice to evaluate the Smo inhibitor, HhAntag, by whole animal functional imaging. HhAntag rapidly reduced systemic luciferase activity in 10- to 14-day-old mice following oral dosing. Although pathway activity was restored 2 days after drug removal, brief inhibition caused permanent defects in bone growth. HhAntag inhibited proliferation and promoted differentiation of chondrocytes, leading to dramatic expansion of the hypertrophic zone. After drug removal, osteoblasts invaded the cartilage plate, mineralization occurred, and there was premature fusion of the growth plate resulting in permanent disruption of bone epiphyses.

本研究では，業界再編が著しい本邦の医薬品産業に着目し，企業の合併・買収（M&A）が医薬研究開発（R&D）活動のイノベーション創出に与える影響を評価し，その企業経営における意義を考察した。手法としては，医薬R&D活動のパフォーマンスを評価のうえ，アステラス製薬の合併事例をケーススタディーに採用し，同社の合併成立後２年時点における合併後マネジメントの発現効果とその背後のメカニズムを，R&D規模，研究効率及びライセンス効率の各指標について調査した。調査の結果，合併後の相互補完による技術機会の拡大がR&D規模に確認され，技術機会の拡大による効率向上がR&D効率及びライセンス効率に有意に認められた。加えて，この統合効果が，同社の掲げる企業ビジョンや合併前に立案された戦略と良く一致していることも確認された。先行研究に基づく通説では，製薬企業のM&Aには開発パイプラインの強化など短期的な効果が認められる一方，永続的な生産性改善には必ずしも寄与しないとされる。本研究は，規模の経済のみならず範囲の経済の視点からの医薬R&D活動の経過観察が，製薬企業の合併後マネジメントの評価に必要であることを示唆するものである。

The sonic hedgehog (Shh) pathway is activated in ∼30% of human medulloblastoma resulting in increased expression of downstream target genes. In about half of these cases, this has been shown to be a consequence of mutations in regulatory genes within the pathway, including Ptc1, Smo, and Sufu. However, for some tumors, no mutations have been detected in known pathway genes. This suggests that either mutations in other genes promote tumorigenesis or that epigenetic alterations increase pathway activity in these tumors. Here, we report that 3% to 4% of mice lacking either one or both functional copies of Cxcr6 develop medulloblastoma. Although CXCR6 is not known to be involved in Shh signaling, tumors derived from Cxcr6 mutant mice expressed Shh pathway target genes including Gli1, Gli2, Ptc2, and Sfrp1, indicating elevated pathway activity. Interestingly, the level of Ptc1 expression was decreased in tumor cells although two normal copies of Ptc1 were retained. This implies that reduced CXCR6 function leads to suppression of Ptc1 thereby increasing Smoothened function and promoting tumorigenesis. We used a direct transplant model to test the sensitivity of medulloblastoma arising in Cxcr6 mutant mice to a small-molecule inhibitor of Smoothened (HhAntag). We found that transplanted tumors were dramatically inhibited in mice treated for only 4 days with HhAntag. These findings suggest that HhAntag may be effective against tumors lacking mutations in known Shh pathway genes. [Cancer Res 2007;67(8):3871–7]

Medulloblastoma is the most common malignant pediatric brain tumor. Current treatment is associated with major long-term side effects; therefore, new nontoxic therapies, targeting specific molecular defects in this cancer, need to be developed. We use a mouse model of medulloblastoma to show that inhibition of the Sonic Hedgehog (Shh) pathway provides a novel therapy for medulloblastoma. A small molecule inhibitor of the Shh pathway, HhAntag, blocked the function of Smoothened in mice with medulloblastoma. This resulted in suppression of several genes highly expressed in medulloblastoma, inhibition of cell proliferation, increase in cell death and, at the highest dose, complete eradication of tumors. Long-term treatment with HhAntag prolonged medulloblastoma-free survival. These findings support the development of Shh antagonists for the treatment of medulloblastoma.

The Sp family is a family of transcription factors that bind to cis-elements in the promoter regions of various genes. Regulation of transcription by Sp proteins is based on interactions between a GC-rich binding site (GGGCGG) in DNA and C-terminal zinc finger motifs in the proteins. In this study, we characterized the GC-rich promoter of the gene for the DNA methyltransferase (Dnmt1) that is responsible for methylation of cytosine residues in mammals and plays a role in gene silencing. We found that a cis-element (nucleotides -161 to -147) was essential for the expression of the mouse gene for Dnmt1. DNA-binding assays indicated that transcription factors Sp1 and Sp3 bound to the same cis-element in this region in a dose-dependent manner. In Drosophila SL2 cells, which lack the Sp family of transcription factors, forced expression of Sp1 or Sp3 enhanced transcription from the Dnmt1 promoter. Stimulation by Sp1 and Sp3 were independent phenomena. Furthermore, cotransfection reporter assays with a p300-expression plasmid revealed the activation of the promoter of the Dnmt1 gene in the presence of Sp3. The transcriptional coactivator p300 interacted with Sp3 in vivo and in vitro. Our results indicate that expression of the Dnmt1 gene is controled by Sp1 and Sp3 and that p300 is involved in the activation by Sp3.

Endothelial Fas ligand (FasL) contributes to the "immune privilege" of tissues such as testis and eye, in which apoptosis is induced in infiltrating Fas-positive activated T cells and results in the inhibition of leukocyte extravasation. In this study, we examined the role of endothelial FasL in controlling cancer cell transmigration using rat lung endothelial (RLE) cell line bearing a doxycycline-inducible hepatocyte nuclear factor (HNF)-4alpha expression system. We showed that a detectable level of FasL was expressed in RLE cells and that this expression was markedly up-regulated and well correlated to the degree of HNF-4alpha expression in a time-dependent manner. When various cancer cells were overlaid on an RLE monolayer sheet, we examined the ability of endothelial FasL to induce massive apoptosis in Fas-expressing cancer cells and found a causal link to inhibition of the transmigration. Finally, we showed that FasL was expressed in capillaries of the rat brain by immunohistochemical staining, suggesting that FasL serves its functions not only in vitro, but also in vivo. These results raise the possibility that HNF-4alpha is involved in regulating cancer cell transmigration by modulating the Fas-FasL system.

How natural selection can promote cooperative or altruistic behavior is a fundamental question in biological and social sciences. One of the persuasive mechanisms is "indirect reciprocity," working through reputation: cooperative behavior can prevail because the behavior builds the donor's good reputation and then s/he receives some reciprocal benefits from someone else in the community. However, an important piece missed in the previous studies is that the reputation-building process requires substantial cognitive abilities such as communication skills, potentially causing a loss of biological fitness. Here, by mathematical analyses and individual-based computer simulations, we show that natural selection never favors indirect reciprocal cooperation in the presence of the cost of reputation building, regardless of the cost-to-benefit ratio of cooperation or moral assessment rules (social norms). Our results highlight the importance of considering the cost of high-level cognitive abilities in studies of the evolution of humans' and animals' social behavior.

7H6 antigen is a novel tight junction-associated protein first identified in our laboratory with a monoclonal antibody generated by immunizing mice with a bile canaliculus-rich membrane fraction obtained from adult rat liver. Although the antigen is found preferentially at the tight junction of epithelial cells in various organs in adult mammals, the relationship between 7H6 antigen and other tight junction proteins remains obscure. In the present study we examined the relationship between the immunohistochemical distribution of 7H6 antigen and occludin in chick intestinal epithelial cells during development in vivo, since occludin is the only known membrane protein composing tight junctions which is found thus far exclusively in avian tissues. 7H6 antigen was found as coarse dots along the basolateral cell membrane of intestinal epithelium in the chick embryo, whereas it was found as a fine dashed line along the cell border in the chicken. In contrast, occludin was found linearly together with ZO-1 at the cell border of intestinal epithelia both in the chick embryo and chicken. The possible significance of such different localization is briefly discussed.

Human brucellosis is an imported zoonosis extremely uncommon in Japan. Brucellosis was found in a 38-year-old surgeon who had never been abroad; he developed intrapulmonary infiltrates and pleural thickening indistinguishable from pulmonary tuberculosis. Despite extensive antibiotic therapy for tuberculosis he developed systemic serositis. A culture of resected lung tissues grew CO2-required coccobacilli. Polymerase chain reaction test using a specific primer pair for Brucella abortus revealed that the isolated pathogen is highly homologous to B. abortus. We emphasize that undetermined cases of human brucellosis may be present in Japan.

Twenty-eight patients received a hepatectomy for hepatic metastases of colorectal cancer at the department from April 1984 to December 1993 were evaluated, especially for H1 patients. We prefered hepatectomy with a surgical margin of more than 1cm. In addition, the continuous infusion therapy with 5-FU (250mg/day) into the hepatic artery was selected as a postoperative adjuvant therapy. Three-year and five-year survivals with H1 patients resulted in 45% and 28%, respectively. None of H2 patients survived more than three years. Recurrence was noted in 43% of H1 patients, and about 90% of the recurrences occurred in the remnant liver after hepatic resection. There was no significant difference between synchronous and metachronous metastases or between males and females. The patients underwent accurate segmentectomy got a favorable prognosis compared with partial hepatectomy. The five-year survival rate of patients undergoing hepatectomy added postoperative chemotherapy combined with continuous hepatic arterial infusion, was 33% which was favorable compared with those added chemotherapy only, without significant. We think that aggressive large resection of the liver including metastatic tumor combined with postoperative chemotherapy using hepatic arterial infusion is an effective treatment of hepatic metastases of colorectal cance for prognostic improvement.

A case of duodenal carcinoid metastasizing to lymph nodes in the posterior region of the pancreatic head, which was difficult to distinguish from a retroperitoneal tumors, is described. A 48-year-old woman was pointed out having a palpable tumor in the right upper abdomen at a periodic examination. Ultrasonography and CT revealed a 7.5cm tumor in the posterior region of the pancreatic head. The patient was admitted to the hospital for further examination. ERCP showed a small elevated lesion, 1.5cm in diameter, with a delle oral-side to the papilla of Vater. From biopsy results a diagnosis of duodenal carcinoma was made. Entertaining a possibility of primal lymph node metastasis or retroperitoneal tumor because of too large lesion as a tumor, existing in the posterior region of the pancreatic head, pancreatoduodenectomy was carried out.In the resected specimen, there were three tumors, namely a 1.5×1.5×1.0cm tumor in the duodenum, and 4.5×5.5×4.5cm and 3.0×2.5×2.5cm tumors at the back of the pancreas. Histopathologically the three tumors were positive for Grimerius staining, and were diagnosed as primary duodenal carcinoid with metastases to the lymph nodes at the back of the pancreas. It is thought that pancreatoduodenectomy with lymph node dissection is necessary even for a duodenal carcinoid of less than 2cm in the maximal diameter.

ゾル-ゲル法によって作製した球状シリカ(比表面積1～700m2/g, 平均粒径4～30μm)のSBR補強性について検討した結果, ゴム補強性に対するシリカ自体の性状要因の一つが明らかになった. それはシリカ微粉体の平衡水分から得られる, 親水性指数(WAI)である. WAIにより, 例えば高比表面積試料での硬さの低下現象を推定することが可能となる. また, 破壊強度については, その吸油量が大きな要因となることがわかった.一方, 高比表面積の球状シリカを市販のシリカ補強剤と比較した結果, 特に圧縮永久ひずみ率の改善の著しいことが認められた.

Abnormal expression of Dnmt1 in vivo induces cellular alterations such as transformation, and an increase in Dnmt1 mRNA plays a causal role in c-fos-, ras- and SV40 large T antigen-induced transformation of fibroblasts in vitro. Here, we have investigated the regulation of Dnmt1 transcription. We identified the promoter region and major transcription start sites of mouse Dnmt1 and found two important cis-elements within the core promoter region. One is an E2F binding site, and the other is a binding site for an as yet unidentified factor. Point mutations in the two cis-elements decreased promoter activity in both non-transformed and transformed cells. Thus, both sites play a critical role in regulation of Dnmt1 transcription in proliferating cells. Treatment with trichostatin A, a specific inhibitor of histone deacetylase, increased Dnmt1 promoter activity in G0/G1-arrested NIH 3T3 cells. Furthermore, the decrease in promoter activity induced by expression of E2F-1 and Rb was reversed by trichostatin A treatment of Saos-2 cells. Taken together, these data indicate that transcription of Dnmt1 is regulated in a complex fashion by E2F and other transcription factors through E2F-Rb-HDAC-dependent and -independent pathways. These findings suggest that Dnmt1 is a target gene of these pathways in cell proliferation, cell transformation and tumorigenesis.

During mammalian cell division, DNA methylation patterns are transferred accurately to the newly synthesized DNA strand. This depends on maintenance DNA methyltransferase activity. DNA methylation can affect chromatin organization and gene expression by recruitment of histone deacetylases (HDACs). Here we show that the methyl-CpG binding protein, MeCP2, interacts directly with the maintenance DNA methyltransferase, Dnmt1. The region of MeCP2 that interacts with Dnmt1 corresponds to the transcription repressor domain which can also recruit HDACs via a corepressor, mSin3A. Dnmt1 can form complexes with HDACs as well as MeCP2. Surprisingly, the MeCP2-Dnmt1 complex does not contain the histone deacetylase, HDAC1. Thus, Dnmt1 takes the place of the mSin3A-HDAC1 complex, indicating that the MeCP2-interacting Dnmt1 does not bind to HDAC1. Further, we demonstrate that MeCP2 can form a complex with hemimethylated as well as fully methylated DNA. Immunoprecipitated MeCP2 complexes show DNA methyltransferase activity to hemimethylated DNA. These results suggest that Dnmt1 associates with MeCP2 in order to perform maintenance methylation in vivo. We propose that genome-wide and/or -specific local DNA methylation may be maintained by the Dnmt1-MeCP2 complexes, bound to hemimethylated DNA. Dnmt1 may be recruited to targeted regions via multiple steps that may or may not involve histone deacetylases.

The Sp family is a family of transcription factors that bind to cis-elements in the promoter regions of various genes. Regulation of transcription by Sp proteins is based on interactions between a GC-rich binding site (GGGCGG) in DNA and C-terminal zinc finger motifs in the proteins. In this study, we characterized the GC-rich promoter of the gene for the DNA methyltransferase (Dnmt1) that is responsible for methylation of cytosine residues in mammals and plays a role in gene silencing. We found that a cis-element (nucleotides -161 to -147) was essential for the expression of the mouse gene for Dnmt1. DNA-binding assays indicated that transcription factors Sp1 and Sp3 bound to the same cis-element in this region in a dose-dependent manner. In Drosophila SL2 cells, which lack the Sp family of transcription factors, forced expression of Sp1 or Sp3 enhanced transcription from the Dnmt1 promoter. Stimulation by Sp1 and Sp3 were independent phenomena. Furthermore, cotransfection reporter assays with a p300-expression plasmid revealed the activation of the promoter of the Dnmt1 gene in the presence of Sp3. The transcriptional coactivator p300 interacted with Sp3 in vivo and in vitro. Our results indicate that expression of the Dnmt1 gene is controled by Sp1 and Sp3 and that p300 is involved in the activation by Sp3.

Expression of rat placental lactogen I is specific to the placenta and never expressed in other tissues. To obtain insight into the mechanism of tissue-specific gene expression, we investigated the methylation status in 3.4 kb of the 5′-flanking region of the rat placental lactogen I gene. We found that the distal promoter region of the rat placental lactogen I gene had more potent promoter activity than that of the proximal area alone, which contains several possible cis-elements. Although there are only 17 CpGs in the promoter region, in vitro methylation of the reporter constructs caused severe suppression of reporter activity, and CpG sites in the placenta were more hypomethylated than other tissues. Coexpression of methyl-CpG-binding protein with reporter constructs elicited further suppression of the reporter activity, whereas treatment with trichostatin A, an inhibitor of histone deacetylase, reversed the suppression caused by methylation. Furthermore, treatment of rat placental lactogen I nonexpressing BRL cells with 5-aza-2′-deoxycytidine, an inhibitor of DNA methylation, or trichostatin A resulted in the de novo expression of rat placental lactogen I. These results provide evidence that change in DNA methylation is the fundamental mechanism regulating the tissue-specific expression of the rat placental lactogen I gene.

Survivin is a member of the inhibitor of apoptosis protein (IAP) family containing a single baculovirus IAP repeat domain. It is expressed during fetal development but becomes undetectable in terminally differentiated normal adult tissues. We previously reported that survivin and its splicing variant survivin-2B was expressed abundantly in various types of tumor tissues as well as tumor cell lines and was suitable as a target antigen for active-specific anti-cancer immunization. Subsequently, we identified an HLA-A24-restricted antigenic peptide, survivin-2B80-88 (AYACNTSTL) recognized by CD8+ cytotoxic T lymphocytes (CTLs). We, therefore, started a phase I clinical study assessing the efficacy of survivin-2B peptide vaccination in patients with advanced or recurrent colorectal cancer expressing survivin. Vaccinations with survivin-2B peptide were given subcutaneously six times at 14-day intervals. Of 15 patients who finished receiving the vaccination schedule, three suffered slight toxicities, including anemia (grade 2), general malaise (grade 1), and fever (grade 1). No severe adverse events were observed in any patient. In 6 patients, tumor marker levels (CEA and CA19-9) decreased transiently during the period of vaccination. Slight reduction of the tumor volume was observed in one patient, which was considered a minor responder. No changes were noted in three patients while the remaining eleven patients experienced tumor progression. Analysis of peripheral blood lymphocytes of one patient using HLA-A24/peptide tetramers revealed an increase in peptide-specific CTL frequency from 0.09% to 0.35% of CD8+ T cells after 4 vaccinations. This phase I clinical study indicates that survivin-2B peptide-based vaccination is safe and should be further considered for potential immune and clinical efficacy in HLA-A24-expression patients with colorectal cancer.

During the last 9 years, 1707 patients with upper abdominal disease underwent surgical treatment at our department. One hundred three patients (6.0%) were infected after primary operation. The incidence of postoperative infection was 8.1% in the A group (upper GI-tract) and 4.4% in the B group (liver, gallbladder, pancreas). Postoperative infection in malignancy occurred at an incidence five times that in benign disease, especially in the B group. Twenty-two patients (21.4%) died from pneumonia, leakage, peritonitis, cholangitis, sepsis and subphrenic abscess. Gram-positive bacillus infection has been an increasingly common postoperative infection during last three years.

Follow-up results of operative treatment for chronic pancreatitis were studied with reference to five indicators, i.e., body weight, effect on relief from pain, pancreatic exocrine and endocrine functions, and social rehabilitation, in 34 patients operated on from June 1974 to May 1984 in our institution. Surgical death did not occur in any cases. The interval observed after operation was at least one year in every patient. Increased body weight was observed in 8 patients (23.5%) especially in the group that underwent drainage operation for dilated puncreatic duct (60%). Some of the patients with alcoholic pancreatitis showed an increase when they avoided the use of alcohol. The effect on the removal of pain by surgical treatment in spite of difficulty with palliative control was significant, except for cystogastrostomy or cystojejunostomy. The decrease in exocrine and endocrine functions after surgical treatment was progressive, and those in the group with resected pancreas were the worst, of three patients whe did not achieve normal social rehabilitation, one had abdominal pain and the other two have continued to drink alcohol. It is considered that the appropriate selection of operative procedures and self-control on the part of the patients are important factors in the development of the results of surgical treatment for chronic pancreatitis.

Though splenic cysts have been considered to be a relatively rare disease, a number of cases have recently been reported corresponding to a marked improvement in diagnostic technique. The clinico-pathological classification of these disease established in the 1940's to 1950's is used at present. However, as clinical experience increases, it has become obvious that there are many unclassifiable cases. A case reported was a eighty five year old woman whose chief complaint was diarrhea. Preoperative diagnosis by angiography, echography and computed tomography suggested a multiple cyst of spleen. At the operation when splenectomy was performed, the other organs seemed macroscopically tobe normal and the size of cystic spleen was 19×10×7cm. Pathological examination of the resected specimen was a mucigenic epithelial cyst of the spleen of a new type beyond the present classification.

People need reliable information regarding over-the-counter medications (OTCs), so that they can independently make appropriate informed choices. The study aimed to identify the information providers and channels that have an impact on the purchase of OTCs, and to demonstrate the information needs of OTC purchasers, using these providers and channels, from the viewpoint of information characteristics such as specialty, objectivity, concreteness, comprehensiveness, individuality, and availability, focusing on the efficacy of OTCs and related safety information. A questionnaire survey of randomly sampled adults aged ≥20 was conducted at the Japan Drugstore Show 2012, hosted by the Japan Association of Chain Drug Stores. In this questionnaire, information was particularly limited to the efficacy and safety of OTCs. Multivariate logistic regression analysis was performed on data from 1743 respondents (1625 purchasers and 118 non-purchasers of OTCs) who obtained information on OTCs in their daily lives, to demonstrate the associations between the use of information providers and channels (predictor variables) and the purchase of OTCs (outcome variable), as well as between information characteristics valued by purchasers (predictor variables) and their use of these information providers or channels (outcome variables). Both the use of pharmacists as information providers and consultation at pharmacies as an information channel were positively associated with the purchase of OTCs (odds ratio [OR], 3.74; 95 % confidence interval [CI], 2.46–5.68; P