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Izumi Ishiyama,Akiko Nagai,Kaori Muto,Akiko Tamakoshi,Minori Kokado,Kyoko Mimura,Tetsuro Tanzawa,Zentaro Yamagata The aim of this study was to assess public attitudes toward the promotion of genomic studies related to medicine and to examine the relationship between public attitudes and the level of genomic literacy by analyzing data from a nationwide opinion survey. The participants comprised 4,000 people (ag... ( view more )e, 20-69) selected from the Japanese general population by using the two-step stratified random sampling method. They were queried on the following topics in a mail survey: (1) pros and cons of the promotion of genomic studies related to medicine, (2) level of scientific literacy in genomics, (3) demographic and socioeconomic background, and (4) knowledge and attitudes toward science in general and genetic testing in particular. We examined the relationship between the approval of promotion and literacy level, using logistic regression models stratified by gender. Our results showed the response rate was 54.3% (2,171/4,000), and 69.4% participants favored the promotion of genomic studies related to medicine. Only 1.3% participants expressed a negative attitude. Multivariate analysis revealed that approval of promotion was related to a high literacy score. This relationship was stronger in males than in females (the highest quartile of score vs. the lowest: adjusted odds ratio, 3.36 for males and 1.86 for females; 95% confidence interval, 1.88-5.98 for males and 1.17-2.95 for females). We determined that a majority of the Japanese participants currently approved of the promotion of genomic studies related to medicine and that people with a high level of genomic literacy tended to approve the promotion. ( view less ) Yoko Komatsu,Ichiaki Ito,Mitsutoshi Wayama,Akiko Fujimura,Kensuke Akaogi,Hikaru Machida,Yuka Nakajima,Takao Kuroda,Kazuji Ohmori,Akiko Murayama,Keiji Kimura,Junn Yanagisawa PPARgamma is a nuclear hormone receptor that plays a key role in the induction of peroxisome proliferation. A number of studies showed that PPARgamma ligands suppress cell cycle progression; however, the mechanism remains to be determined. Here, we showed that PPARgamma ligand troglitazone inhibite... ( view more )d G1/S transition in colon cancer cells, LS174T. Troglitazone did not affect on either expression of CDK inhibitor (p18) or Wnt signaling pathway, indicating that these pathways were not involved in the troglitazone-dependent cell cycle arrest. GeneChip and RT-PCR analyses revealed that troglitazone decreased mRNA levels of cell cycle regulatory factors E2F2 and cyclin-E1 whose expression is activated by E2F2. Down-regulation of E2F2 by troglitazone results in decrease of cyclin-E1 transcription, which could inhibit phosphorylation of Rb protein, and consequently evoke the suppression of E2F2 transcriptional activity. Thus, we propose that troglitazone suppresses the feedback loop containing E2F2, cyclin-E1, and Rb protein. ( view less ) Akiko Murayama,Kazuji Ohmori,Akiko Fujimura,Hiroshi Minami,Kayoko Yasuzawa-Tanaka,Takao Kuroda,Shohei Oie,Hiroaki Daitoku,Mitsuru Okuwaki,Kyosuke Nagata,Akiyoshi Fukamizu,Keiji Kimura,Toshiyuki Shimizu,Junn Yanagisawa Intracellular energy balance is important for cell survival. In eukaryotic cells, the most energy-consuming process is ribosome biosynthesis, which adapts to changes in intracellular energy status. However, the mechanism that links energy status and ribosome biosynthesis is largely unknown. Here, w... ( view more )e describe eNoSC, a protein complex that senses energy status and controls rRNA transcription. eNoSC contains Nucleomethylin, which binds histone H3 dimethylated Lys9 in the rDNA locus, in a complex with SIRT1 and SUV39H1. Both SIRT1 and SUV39H1 are required for energy-dependent transcriptional repression, suggesting that a change in the NAD(+)/NADH ratio induced by reduction of energy status could activate SIRT1, leading to deacetylation of histone H3 and dimethylation at Lys9 by SUV39H1, thus establishing silent chromatin in the rDNA locus. Furthermore, eNoSC promotes restoration of energy balance by limiting rRNA transcription, thus protecting cells from energy deprivation-dependent apoptosis. These findings provide key insight into the mechanisms of energy homeostasis in cells. ( view less ) Masanori Tohno,Tomoyuki Shimazu,Hisashi Aso,Akiko Uehara,Haruhiko Takada,Akiko Kawasaki,Yukari Fujimoto,Koichi Fukase,Tadao Saito,Haruki Kitazawa In this study, we isolated a complementary DNA encoding nucleotide-binding oligomerization domain-1 (NOD1) from Peyer's patches (Pps) of swine gut-associated lymphoid tissues (GALT). The complete open reading frame of porcine NOD1 contains 2862 bp, encoding a 953-amino acid polypeptide. The porcine... ( view more ) NOD1 amino acid sequence is more closely related to the human sequence (83.8% identity) than the mouse counterpart (79.2% identity). To examine the subcellular expression and function of porcine NOD1, we overexpressed it in human embryonic kidney 293 cells. Immunostaining with an anti-porcine NOD1 polyclonal antibody revealed that the protein was expressed in transfectants as an intracellular membrane-bound molecule. In the transfected cells, both gamma-d-glutamyl-meso-diaminopimelic acid, and meso-diaminopimelic acid and meso-lanthionine activated nuclear factor-kappa B. Quantitative real-time PCR detected NOD1 mRNA in multiple tissues isolated from adult and newborn swine, including the esophagus, duodenum, jejunum, ileum, ileal Pps, colon, spleen, and mesenteric lymph nodes. In the newborn and adults, NOD1 was highly expressed in the esophagus and GALT, such in the ileal Pps and mesenteric lymph nodes. Furthermore, Toll-like receptor and NOD1 ligands as well as immunobiotic lactic acid bacteria enhanced the expression of NOD1 in GALT of adult and newborn swine. Our results should help clarify how the intestinal immune system is modulated by low-molecular weight peptidoglycan fragments through NOD1. ( view less ) Nana Mukai,Taichi Akahori,Motohiro Komaki,Qin Li,Toshie Kanayasu-Toyoda,Akiko Ishii-Watabe,Akiko Kobayashi,Teruhide Yamaguchi,Mayumi Abe,Teruo Amagasa,Ikuo Morita The identification of circulating endothelial progenitor cells (EPCs) has revolutionized approaches to cell-based therapy for injured and ischemic tissues. However, the mechanisms by which EPCs promote the formation of new vessels remain unclear. In this study, we obtained early EPCs from human per... ( view more )ipheral blood and late EPCs from umbilical cord blood. Human umbilical vascular endothelial cells (HUVECs) were also used. Cells were evaluated for their tube-forming potential using our novel in vitro assay system. Cells were seeded linearly along a 60 mum wide path generated by photolithographic methods. After cells had established a linear pattern on the substrate, they were transferred onto Matrigel. Late EPCs formed tubular structures similar to those of HUVECs, whereas early EPCs randomly migrated and failed to form tubular structures. Moreover, late EPCs participate in tubule formation with HUVECs. Interestingly, late EPCs in Matrigel migrated toward pre-existing tubular structures constructed by HUVECs, after which they were incorporated into the tubules. In contrast, early EPCs promote sprouting of HUVECs from tubular structures. The phenomena were also observed in the in vivo model. These observations suggest that early EPCs cause the disorganization of pre-existing vessels, whereas late EPCs constitute and orchestrate vascular tube formation. ( view less ) Takakazu Kaneko,Nobuyoshi Nakajima,Shinobu Okamoto,Iwane Suzuki,Yuuhiko Tanabe,Masanori Tamaoki,Yasukazu Nakamura,Fumie Kasai,Akiko Watanabe,Kumiko Kawashima,Yoshie Kishida,Akiko Ono,Yoshimi Shimizu,Chika Takahashi,Chiharu Minami,Tsunakazu Fujishiro,Mitsuyo Kohara,Midori Katoh,Naomi Nakazaki,Shinobu Nakayama,Manabu Yamada,Satoshi Tabata,Makoto M Watanabe The nucleotide sequence of the complete genome of a cyanobacterium, Microcystis aeruginosa NIES-843, was determined. The genome of M. aeruginosa is a single, circular chromosome of 5,842,795 base pairs (bp) in length, with an average GC content of 42.3%. The chromosome comprises 6312 putative prote... ( view more )in-encoding genes, two sets of rRNA genes, 42 tRNA genes representing 41 tRNA species, and genes for tmRNA, the B subunit of RNase P, SRP RNA, and 6Sa RNA. Forty-five percent of the putative protein-encoding sequences showed sequence similarity to genes of known function, 32% were similar to hypothetical genes, and the remaining 23% had no apparent similarity to reported genes. A total of 688 kb of the genome, equivalent to 11.8% of the entire genome, were composed of both insertion sequences and miniature inverted-repeat transposable elements. This is indicative of a plasticity of the M. aeruginosa genome, through a mechanism that involves homologous recombination mediated by repetitive DNA elements. In addition to known gene clusters related to the synthesis of microcystin and cyanopeptolin, novel gene clusters that may be involved in the synthesis and modification of toxic small polypeptides were identified. Compared with other cyanobacteria, a relatively small number of genes for two component systems and a large number of genes for restriction-modification systems were notable characteristics of the M. aeruginosa genome. ( view less ) Saemi Kato,Akiko Shimizu-Ibuka,Kiyoshi Mura,Akiko Takeuchi,Chiyoko Tokue,Soichi Arai An alpha-amylase secreted by Pichia burtonii 15-1 isolated from a traditional starter murcha of Nepal, named Pichia burtonii alpha-amylase (PBA), was studied. The gene was cloned and its nucleotide sequence was determined. PBA was deduced to consist of 494 amino acid residues. It shared certain deg... ( view more )rees of amino acid sequence identity with other homologous proteins: 60% with Schwanniomyces occidentalis alpha-amylase, 58% with Saccharomycopsis sp. alpha-amylase, and 47% with Taka-amylase A from Aspergillus oryzae. A three-dimensional structural model of PBA generated using the known three-dimensional structure of Taka-amylase A as a template suggested high structural similarity between them. Kinetic analysis revealed that the K(m) values of PBA were lower than those of Taka-amylase A for the oligosaccharides. Although the k(cat) values of PBA were lower than those of Taka-amylase A for the oligosaccharide substrates, the k(cat)/K(m) values of PBA were higher. ( view less ) Minako Mori,Hayato Maruoka,Yuya Nagai,Haruhiko Fujita,Katsuhiro Togami,Sumie Tabata,Masahiro Kurata,Akiko Matsushita,Kenichi Nagai,Kyoko Tanaka,Akiko Yamashiro,Takayuki Takahashi A 80-year-old man was admitted because of acute-onset thrombocytopenia and renal failure. He was diagnosed with Bence Jones (lambda) -type multiple myeloma associated with sepsis with methicillin-resistant Staphylococcus aureus. On admission, serum amylase activity was elevated to 1,814 IU/l (98% s... ( view more )alivary type; S-amylase). Several days after admission, he developed bilateral myelomatous pleuritis. The activity of S-amylase in the effusion was 5,495 IU/l. Myeloma cells in the pleural effusion were positive for cytoplasmic amylase with an antibody against human amylase. High S-amylase activity was detected in the supernatant of cultured myeloma cells in the effusion. Furthermore, S-amylase gene expression was detected by RT-PCR. A diagnosis of amylase-producing multiple myeloma was made. The patient died of renal insufficiency complicated by severe DIC. We report a rare case of amylase-producing myeloma confirmed by immunocytochemistry, culture method, and gene expression. ( view less ) Akiko M Saito,Masahiro Kami,Shin-Ichiro Mori,Yoshinobu Kanda,Ritsuro Suzuki,Shin Mineishi,Akiyoshi Takami,Shuichi Taniguchi,Yoshinobu Takemoto,Masamichi Hara,Masaki Yamaguchi,Masayuki Hino,Takashi Yoshida,Sung-Won Kim,Akiko Hori,Yasuo Ohashi,Yoichi Takaue This prospective trial assessed the safety and efficacy of allogeneic hematopoietic stem cell transplantation from a HLA-matched donor with a reduced-intensity regimen (RIST) consisting of iv fludarabine 30 mg/m(2) for 6 days and oral busulfan 4 mg/kg/day for 2 days in patients older than 50 years ... ( view more )with hematological malignancies. Cyclosporine alone or cyclosporine with short-term methotrexate was randomized for graft-versus-host disease prophylaxis. After 30 patients had been enrolled, an interim analysis was performed, and this report focuses on a precise evaluation of the toxicity profile and chimerism kinetics. Sustained engraftment in all patients, no severe regimen-related toxicity (RRT) within 20 days, and no transplant-related mortality through Day 100 were observed. T-cell (CD3+) full-donor (over 90%) chimerism was observed in 22 of the 30 patients, while the remaining eight had mixed-donor chimerism over 77% on Day 90. Thereafter, five subsequently converted to full-donor chimerism without donor lymphocyte infusion by day 120 (n = 4) or Day 180 (n = 1). Two showed persistent mixed chimerism without relapse through Day 180. Grade III-IV acute graft-versus-host disease and extensive chronic graft-versus-host disease occurred in 10% and 73%, respectively. With a median follow-up of 1.5 years, overall survival and disease-free survival at 1 year was 83% and 62%, respectively. Seven patients hematologically relapsed overall, and five of them had myelodysplastic syndrome with poor prognostic factors. In older patients, RIST with fludarabine and busulfan was associated with acceptable toxicities and a satisfactory antileukemia effect, regardless of the early chimerism status. ( view less ) Ryosuke Osada,Akiko Horiuchi,Norihiko Kikuchi,Junko Yoshida,Akiko Hayashi,Masao Ota,Yoshihiko Katsuyama,Giovanni Melillo,Giovanni Mellilo,Ikuo Konishi The hypoxia-inducible factor (HIF) is a transcriptional factor with important roles in tumor biology. To clarify the possible involvement of the HIF-alpha subunit and von Hippel-Lindau (VHL) protein in the development and progression of ovarian carcinoma, we analyzed the immunohistochemical express... ( view more )ions of HIF-1alpha, HIF-2alpha, and VHL in 107 cases of epithelial ovarian tumors. In addition, we examined loss of heterozygosity (LOH) at VHL gene loci. The frequency of the cytoplasmic expression of HIF-2alpha in carcinomas was higher than that in benign and borderline tumors (P < .0001). Furthermore, the nuclear expression of HIF-1alpha and the cytoplasmic expression of HIF-2alpha were significantly higher in tumors of FIGO (International Federation of Gynecology and Obstetrics) stages III and IV than in those of stages I and II. On the other hand, the cytoplasmic expression of HIF-1alpha did not show differences among histological malignancies. There was a positive correlation between nuclear HIF-1alpha expression and vascular endothelial growth factor (rho = 0.320, P < .001). Although LOH at the VHL gene locus was frequent in ovarian carcinomas (24%), there is no significant correlation between LOH and loss of VHL expression. In 22 clear cell carcinomas, VHL expression showed a significantly negative correlation with the nuclear expression of HIF-1alpha (rho = -0.529, P = .0153). The log-rank test showed that nuclear positive immunostaining for HIF-1alpha (P = .002) and cytoplasmic positive immunostaining for HIF-2alpha (P = .0112) in tumor cells are associated with poor prognosis of patients with ovarian carcinoma. Multivariate analysis also showed that the nuclear expression of HIF-1alpha is an independent prognostic factor. These results show that the HIF-alpha subunit represents an important biomarker in the evaluation of the prognosis of patients with ovarian carcinoma. ( view less ) Satoko Hori,Nobuko Matsuo,Akiko Yamamoto,Tomomi Hazui,Hiromi Yagi,Marumi Nakano,Yuka Suzuki,Akiko Miki,Hisakazu Ohtani,Yasufumi Sawada AIMS: To present a case of piloerection after replacing fluvoxamine maleate with milnacipran hydrochloride, and to analyse this effect based on receptor occupancy theory. METHODS: A 40-year-old female with a 3-year history of panic disorder was prescribed fluvoxamine 50 mg day(-1) in addition to cl... ( view more )orazepate dipotassium and sulpiride. Depression was not improved and she complained of fatigue, lack of energy and drowsiness. These symptoms worsened within a few days of an increase in the dose of fluvoxamine to 50 mg twice daily. Since an interaction between fluvoxamine and tizanidine, prescribed by another clinic, was suspected, fluvoxamine was replaced with milnacipran 50 mg day(-1). Although her drowsiness improved, she complained of piloerection throughout her body. This symptom gradually abated within a week and when the dosage of milnacipran was increased to 100 mg day(-1) at 2 months, no further piloerection occurred. We calculated the changes in alpha(1)-adrenoceptor occupancy by endogenous norepinephrine during treatment with the usual doses of milnacipran, fluvoxamine and imipramine by using pharmacokinetic and pharmacodynamic parameters obtained from the literature. RESULTS: The ratios of alpha(1)-adrenoceptor occupancy by endogenous norepinephrine during the treatment with milnacipran, fluvoxamine and imipramine to that without drug were estimated to be 7.13, 1.00 and 4.12, respectively. The alpha(1)-adrenoceptor occupancy by endogenous norepinephrine was increased in a dose-dependent manner by milnacipran, whereas fluvoxamine had essentially no effect. CONCLUSIONS: The piloerection observed after the replacement of fluvoxamine with milnacipran in this patient appears to have been due to an increase in the alpha(1)-adrenoceptor occupancy by endogenous norepinephrine induced by milnacipran. ( view less ) Yukitaka Morita,Hiroshi Ujike,Yuji Tanaka,Kyohei Otani,Makiko Kishimoto,Akiko Morio,Tatsuya Kotaka,Yuko Okahisa,Masayuki Matsushita,Akiko Morikawa,Kenji Hamase,Kiyoshi Zaitsu,Shigetoshi Kuroda BACKGROUND: Serine racemase (SRR) is a brain-enriched enzyme that converts L-serine to D-serine, which acts as an endogenous ligand of N-methyl D-aspartate (NMDA) receptors. Dysfunction of SRR may reduce the function of NMDA receptors and susceptibility to schizophrenia. METHODS: We genotyped three... ( view more ) single-nucleotide polymorphisms (SNPs) of the 5' region of the SRR gene in 525 patients with schizophrenia and 524 healthy controls. Effects of SNPs on the promoter activity and on serum levels of total and D-serine were examined. RESULTS: We found a significant excess of the IVS1a+465C allele of the SRR gene in schizophrenia, especially in the paranoid subtype (p = .0028). A reporter assay showed that the IVS1a+465C allele had 60% lower promoter activity than did the IVS1a+465G allele. CONCLUSIONS: The IVS1a+465C allele of the SRR gene, which reduces expression of the gene, is a risk factor for schizophrenia, especially the paranoid subtype. ( view less ) Akiko Inagaki,Shinichi Yatsuda,Shinichi Edure,Akiko Suzuki,Takeshi Takahashi,Munetaka Akita Mononuclear ruthenium complexes and dinuclear Ru...Pd complexes having a series of 2,2'-bipyrimidine ligands, [(bpy)2Ru(Ln)]2+ [Ln = 2,2'-bipyrimidine (L1), 5,5'-dimethyl-2,2'-bipyrimidine (L2), 5,5'-dibromo-2,2'-bipyrimidine (L3), 4,4'-dimethyl-2,2'-bipyrimidine (L4), and 4,4',6,6'-tetramethyl- 2,... ( view more )2'-bipyrimidine (L5)] and [(bpy)2Ru(Ln)PdL]m+ [Ln = L1-L3; PdL = PdMeCl (m = 2) and PdMe(solvent) (m = 3)], are prepared, and the obtained complexes are characterized by means of spectroscopic and crystallographic methods. Introduction of the substituents on the bipyrimidine ligands led to the substantial differences in their electrochemical and photophysical properties. Density functional theory calculations have been performed to understand the substituent effect on the ground-state molecular orbital energy level. Reactivity studies on the catalytic dimerization of alpha-methylstyrene revealed that the Pd complex having a Br-substituted bipyrimidine ligand were much more active than those of the corresponding Pd complexes having methyl-substituted or nonsubstituted bipyrimidine ligands. ( view less ) Ayumi Fujinaga,Yutaka Fukushima,Akiko Kojima,Yoshikazu Sai,Yoshifumi Ohashi,Akiko Kuzukawa,Tomoyoshi Seto,Shuichi Nosaka We present the case of a morbidly obese woman, with a body mass index (BMI) of 73.7 kg.m(-2), who had a gynecological operation under combined general and epidural anesthesia. The patient's trachea was intubated, using a fiberscope, while she was breathing spontaneously after the intravenous inject... ( view more )ion of fentanyl and propofol as sedatives. Anesthesia was maintained with intravenous propofol and epidural mepivacaine. When the gynecologist placed a sponge in the abdominal cavity to retract the bowel, the patient experienced severe arterial deoxygenation and mild hypotension, due to massive atelectasis of the left lung. Both oxygenation and perfusion were corrected by the removal of the sponge and with the placement of a pillow under the patient's left shoulder. The atelectasis resulted from compression of the left lung by the fatty mediastinum and by the diaphragm being pushed up by the sponge. The hypotension resulted from impaired venous return and hypoxia. The patient suffered no perioperative complications other than atelectasis and a surgical-site infection. Key factors that contributed to the favorable outcome of this patient included a careful tracheal intubation technique, the choice and dose of anesthetic agents, immediate correction of the factors leading to atelectasis, early ambulation, and prophylaxis for deep vein thrombosis. ( view less ) Masamichi Kuwajima,Hiroaki Fujihara,Hiroyoshi Sei,Asako Umehara,Masako Sei,Tomi T Tsuda,Akiko Sukeno,Tatsuya Okamoto,Akiko Inubushi,Yoichi Ueta,Toshio Doi,Hiroshi Kido The mechanism of onset of hypoglycemia in patients with carnitine deficiency has yet to be determined. Using mice with systemic carnitine deficiency (JVS mice), we examined this mechanism, focusing on the weaning period (days 14-28 postpartum). For normal mice, the survival rate was 100%, and no hy... ( view more )poglycemia was observed at all. Gastric lactose began to decrease on day 17, and cellulose increased sharply in amount thereafter. For JVS mice, the survival rate was 77% on day 14 and 28% on day 28. From day 21 on, hypoglycemia was noted. Gastric lactose had disappeared almost completely by day 17, and cellulose was almost undetectable from days 14 to 28. Expression of orexin mRNA in the hypothalamus did not differ between JVS and normal mice on day 14, but was suppressed in JVS mice on days 21 and 28. When JVS mice were fed a carnitine-rich diet, suppression of expression of orexin mRNA in hypothalamus was eliminated, and on day 28 lactose and cellulose were detected in the stomach without hypoglycemia. In conclusion, the suppression of the expression of orexin in the hypothalamus during the weaning period may be involved in the marked anorexia in JVS mice, which eventually leads to death from hypoglycemia. ( view less ) Masaaki Oda,Akiko Yamagiwa,Shinji Yamamoto,Takao Nakayama,Akiko Tsumura,Hiroshi Sasaki,Kazuki Nakao,En Li,Masaki Okano DNA methylation is a major epigenetic mechanism that has been suggested to control developmental gene regulation during embryogenesis, but its regulatory mechanisms remain unclear. In this report, we show that CpG islands associated with the X-linked homeobox gene cluster Rhox, which is highly expr... ( view more )essed in the extraembryonic trophectoderm, are differentially methylated in a stage- and lineage-specific manner during the post-implantation development of mice. Inactivation of both Dnmt3a and Dnmt3b, DNA methyltransferases essential for the initiation of de novo DNA methylation, abolished the establishment of DNA methylation and the silencing of Rhox cluster genes in the embryo proper. The Dnmt3-dependent CpG-island methylation at the Rhox locus extended for a large genomic region ( approximately 1 Mb) containing the Rhox cluster and surrounding genes. Complementation experiments using embryonic stem (ES) cells deficient in the DNA methyltransferases suggested that the CpG-island methylation by Dnmt3a and Dnmt3b was restricted within this large genomic region, and did not affect the neighboring genes outside it, implicating the existence of region-specific boundaries. Our results suggest that DNA methylation plays important roles in both long-range gene silencing and lineage-specific silencing in embryogenesis. ( view less ) Akiko Takeuchi,Akiko Shimizu-Ibuka,Yoshitaka Nishiyama,Kiyoshi Mura,Sanae Okada,Chiyoko Tokue,Soichi Arai Among more than 20 yeast strains isolated from the traditional starter "murcha" in Nepal, we characterized a yeast that might be involved in saccharification. This strain, identified as Pichia burtonii, produced an extracellular amylolytic enzyme when cultured in the presence of starch in the mediu... ( view more )m. Since no amylase secreted by P. burtonii has yet been reported, we purified the enzyme and determined its N-terminal amino acid sequence. Together with the results of a hydrolyzing activity assay toward various substrates, it was found to be an alpha-amylase. The purified enzyme, named Pichia burtonii alpha-amylase (PBA), was a glycoprotein with an apparent molecular mass of 51 kDa. Enzyme activity was optimal at pH 5.0 at 40 degrees C. The enzyme retained 80% of its original activity after incubation under the optimal pH condition at 50 degrees C for 30 min. The activity was inhibited by metal ions such as Cd(2+), Cu(2+), Hg(2+), Al(3+), and Zn(2+). ( view less ) Reizo Kato,Akiko Tajima,Akiko Nakao,Masafumi TamuraPressure-induced superconducting behavior was found in two anion radical salts, EtMe3Z[Pd(dmit)2]2 (dmit = 1,3-dithiole-2-thione-4,5-dithiolate, Z = P, As), that are Mott insulators and exhibit different magnetic and structural transitions at ambient pressure. Akiko Uehara,Yukari Fujimoto,Akiko Kawasaki,Shoichi Kusumoto,Koichi Fukase,Haruhiko Takada Peptidoglycans (PGNs) are ubiquitous constituents of bacterial cell walls and exhibit various immunobiological activities. Two types of minimum essential PGN structures for immunobiological activities were chemically synthesized and designated as muramyldipeptide; N-acetylmuramyl-L-alanyl-D-isoglut... ( view more )amine (MDP) and gamma-D-glutamyl-meso-diaminopimelic acid (iE-DAP), which are common constituents of both Gram-positive and Gram-negative bacteria, as well as most Gram-negative and some Gram-positive bacteria, respectively. Recently, intracellular receptors for MDP and iE-DAP have been demonstrated to be nucleotide-binding oligomerization domain (NOD)1 and NOD2, respectively. In this study, we demonstrated that chemically synthesized meso-DAP itself activated human epithelial cells from various tissues, through NOD1 to generate antibacterial factors, PGN recognition proteins and beta-defensin 2, and cytokines in specified cases, although the activities of meso-DAP were generally weaker than those of known NOD agonists. However, stereoisomers of meso-DAP, LL-DAP, and DD-DAP were only slightly activated or remained inactive, respectively. Synthetic meso-lanthionine, which is another diamino-type amino acid specific to PGN of the specified Gram-negative bacteria, was also recognized by NOD1. In human monocytic cells, in the presence of cytochalasin D meso-DAP induced slightly but significantly increased production of cytokines, although the cells did not respond to meso-DAP in the absent of cytochalasin D. Our findings suggest that NOD1 is a special sentinel molecule, especially in the epithelial barrier, allowing the intracellular detection of bacteria through recognizing meso-DAP or comparable moiety of PGN from specified bacteria in cooperation with NOD2, thereby playing a key role in innate immunity. ( view less ) Tsuyoshi Otsuji,Akiko Tsumura,Kanji Takahashi,Kenichiro Sho,Yoshimi Nagai,Toshio Fukuchi,Akiko Arisawa,Miyo Matsumura BACKGROUND: Some cases of polypoidal choroidal vasculopathy (PCV) in their natural course develop into classic choroidal neovascularization(CNV) as shown by fluorescein angiography (FA) findings. SUBJECTS AND METHOD: We evaluated 8 eyes of 8 PCV patients showing classic CNV by FA findings, using in... ( view more )docyanine green angiography (IA) and optical coherence tomography(OCT). RESULT: All patients showed subretinal grayish exudates, which were considered fibrinous. Five cases were recognized as true subretinal CNV according to IA and OCT findings. The other 3 patients showed polypoidal dilatation with vascular networks by IA, and a moderately reflective mass considered fibrinous over the polypoidal elevation of retinal pigment epithelium (RPE) by OCT. CONCLUSION: Both true CNV and PCV with fibrin are present in PCV patients showing classic CNV. It requires care to determine proper treatment. ( view less ) Shinya Uchino,Shiro Noguchi,Hiroto Yamashita,Hiroyuki Yamashita,Shin Watanabe,Takahiro Ogawa,Akiko Tsuno,Akiko Murakami,Akira Miyauchi INTRODUCTION: Familial adenomatous polyposis (FAP) is an inherited autosomal dominant syndrome caused by germline mutations in the adenomatous polyposis coli (APC) gene. Papillary thyroid cancer is one of the extracolonic manifestations of FAP. A characteristic histologic feature of this type of th... ( view more )yroid tumor is the cribriform-morula variant of papillary thyroid carcinoma (CMVPTC). METHODS: To investigate roles of the APC and beta-catenin genes in the development of CMVPTC, we examined germline and somatic mutations of these genes in a female patient with CMVPTC and FAP. The patient had undergone total colectomy at the age of 19 years and total thyroidectomy at age 25 years. RESULTS: Numerous tumors were disseminated in both lobes of the thyroid gland, and histopathologic examination revealed typical CMVPTC. DNA was extracted from peripheral blood leukocytes and 12 CMVPTC tumors, and exons 1-15 of the APC gene and exon 3 of the beta-catenin gene were examined. A germline mutation was detected in exon 13 of the APC gene, and this mutation generated a premature stop codon. Six somatic mutations (922delC, 1602delA, 1821delT, 1920delG, 2706del20, 2804insA) were found in the CMVPTC specimens. All mutations were truncating mutations in the N-terminus of the APC protein. Loss of heterozygosity was not observed in the remaining tumor tissues without somatic APC mutations. There were no mutations of the beta-catenin gene in peripheral blood leukocytes or 12 CMVPTC specimens. CONCLUSIONS: These results suggest that APC mutations play an important role in the development of CMVPTC and occur predominantly in the 5' side of the APC gene between codons 308 and 935. ( view less ) Akiko Hashimoto Satoi,Kohei Murao,Kozue Kubo,Akiko Takeyasu,Atsushi Ohashi,Shinichi Nakao,Koh Shingu The need for one-lung ventilation has been increasing even in pediatric patients. However, the trachea is so narrow in pediatric patients that ordinary double-lumen tubes can not be used and there have been many reports on devices or measures to block one lung. We report our experience with a femal... ( view more )e infant weighing 2 kg who had severe chronic lung disease under mechanical ventilation, and underwent left lung lower lobectomy with one-lung ventilation technique. We chose a balloon wedge pressure catheter to block the left main bronchus, because it has a central lumen through which a guide wire can be passed and sucking is available. The infant was in need of continuous ventilation and the catheter was too soft to be inserted directly. We first inserted an 18G catheter of a needle-catheter assemble outside the tracheal tube through which a guide wire was inserted into the left main bronchus with the aid of direct vision of a 2-mm fiberoptic bronchoscope through the tracheal tube, and then inserted the balloon wedge pressure catheter placing it in an appropriate position. One-lung ventilation was successfully achieved and the operative and postoperative course was uneventful. ( view less ) Namthip Chumpolkulwong,Kensaku Sakamoto,Akiko Hayashi,Fumie Iraha,Naoko Shinya,Natsuko Matsuda,Daisuke Kiga,Akiko Urushibata,Mikako Shirouzu,Kenji Oki,Takanori Kigawa,Shigeyuki Yokoyama We analyzed the effect of nine 'rare' codons (AGA, AGG, AUA, CCC, CGA, CGG, CUA, GGA, and UUA) on gene expression in an Escherichia coli coupled transcription/translation cell-free system, in comparison with a cell-based expression system. Each reporter gene contained five consecutive repeats of a ... ( view more )rare codon, or in some experiments, three consecutive repeats. The cell-free expression of the genes bearing the codons CGA, CUA, GGA, and UUA was not affected, although these codons, except for GGA, were inefficiently translated in E. coli cells. Translation of the remaining five codons (AGA, AGG, AUA, CCC, and CGG) was severely reduced in both systems, and was remarkably facilitated in the cell-free system based on an S30 extract from the E. coli cells overproducing 'minor' tRNAs for these codons. ( view less ) Yasuhiro Tanaka,Masayuki Kurata,Katsuhiro Togami,Haruyuki Fujita,Naoko Watanabe,Akiko Matsushita,Akinori Maeda,Kenichi Nagai,Akiko Sada,Toshimitsu Matsui,Takayuki Takahashi Hypereosinophilic syndrome (HES) was diagnosed in December 2000 in a 43-year-old man on the basis of persistent eosinophilia (11.7 x 10(9)/L) and a normal karyotype of the bone marrow cells. He had developed intra-abdominal non-Hodgkin's lymphoma and in 1992 had received 3 courses of combination ch... ( view more )emotherapy with doxorubicin (Adriamycin), cyclophosphamide, vincristine, methotrexate, bleomycin, and prednisolone. The patient was orally given prednisolone (10 mg/day) and cyclophosphamide (50 mg/day) as HES treatment without a subsequent improvement of the eosinophilia. In May 2003, anemia (hemoglobin, 7.9 g/dL) and thrombocytopenia (65 x 10(9)/L) manifested with progressive eosinophilia (21.0 x 10(9)/L) and a small number of blasts. The patient became febrile and was admitted in July 2003. Cytogenetic reexamination of the bone marrow cells disclosed the deletion of 4q12, indicating the presence of a fusion of the Fip1-like 1 (FIP1L1) gene to the plateletderived growth factor receptor alpha (PDGFRalpha) gene and consequently the clonal nature of his hematopoietic cells. DNA sequence analysis demonstrated that the breakpoints of the FIP1L1 and PDGFRalpha genes were present in exon 9 and exon 12, respectively. Treatment with imatinib mesylate (300 mg/day) promptly brought about complete remission. Although a number of similar eosinophilic cases have been reported, our patient may be the first such patient with a history of chemotherapy. ( view less ) Sumio Maeda,Miki Ito,Tsuyako Ando,Yuko Ishimoto,Yukiko Fujisawa,Hitomi Takahashi,Akiko Matsuda,Akiko Sawamura,Shiho Kato We tested the possibility of nonconjugative lateral DNA transfer in a colony biofilm of mixed Escherichia coli strains. By simply coculturing a plasmid-free F(-) strain and another F(-) strain harboring a nonconjugative plasmid in a colony biofilm on antibiotic-free agar media, transformed cells we... ( view more )re produced within 24-48 h at the frequency of 10(-10)-10(-9) per recipient cell. PCR analysis of the transformed cells demonstrated the occurrence of lateral plasmid transfer. These cells survived until at least day 7 under antibiotic-free conditions. Liquid cultures of the same strains in Luria-Bertani broth produced no or few transformants, suggesting the importance of colony-biofilm formation for plasmid transfer. This is a novel line of evidence indicating that nonconjugative, nonviral horizontal gene transfer can occur between E. coli cells. ( view less )
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