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A 25-year-old male (Case 1) was waiting for a bone marrow transplant for myelodysplastic syndrome. Due to acute appendicitis, he was advised to undergo gastroenterological surgery. After blood transfusion, he underwent an emergency laparoscopic appendectomy, as no blood cell recovery was expected. The postoperative course was uneventful, and he was discharged. A 71-year-old female (Case 2) developed acute appendicitis during chemotherapy for acute myeloid leukemia (AML). At the time of onset, since her myelosuppression was expected to improve in approximately 1 week, a conservative treatment was administered. However, due to the progression of AML, the expected blood cell recovery did not occur. Therefore, laparoscopic appendectomy was performed 25 days after onset. She was discharged without postoperative adverse events. In cases of acute appendicitis in patients with hematologic disease accompanied by pancytopenia, it is important to establish a careful treatment plan considering the possibility of recovery from myelosuppression and the need to control an intraperitoneal infection in conjunction with a hematologist. Laparoscopic surgery, which is minimally invasive, was an effective surgical procedure.

Aims/Introduction Although genome-wide association studies have identified more than 50 susceptibility genes for type 1 diabetes, low-frequency risk variants could remain unrecognized. The present study aimed to identify novel type 1 diabetes susceptibility genes by newly established methods. Materials and Methods We performed whole-exome sequencing and genome-wide copy-number analysis for a Japanese family consisting of two patients with type 1 diabetes and three unaffected relatives. Further mutation screening was carried out for 127 sporadic cases. The functional consequences of identified substitutions were evaluated by in silico analyses and fluorescence-activated cell sorting of blood samples. Results Whole-exome sequencing and genome-wide copy-number analysis of familial cases showed co-segregation of the p.V863L substitution in CD101, the human homolog of an autoimmune diabetes gene in the nonobese diabetic mouse, with type 1 diabetes. Mutation screening of CD101 in 127 sporadic cases detected the p.V678L and p.T944R substitutions in two patients. The p.V863L, p.V678L, and p.T944R substitutions were absent or extremely rare in the general population and were assessed as “probably/possibly damaging” by in silico analyses. CD101 expression on monocytes, granulocytes, and myeloid dendritic cells of mutation-positive patients was weaker than that of control individuals. Conclusions These results raise the possibility that CD101 is a susceptibility gene for type 1 diabetes. This article is protected by copyright. All rights reserved.

Whole-genome and -exome resequencing using next-generation sequencers is a powerful approach for identifying genomic variations that are associated with diseases. However, systematic strategies for prioritizing causative variants from many candidates to explain the disease phenotype are still far from being established, because the population-specific frequency spectrum of genetic variation has not been characterized. Here, we have collected exomic genetic variation from 1208 Japanese individuals through a collaborative effort, and aggregated the data into a prevailing catalog. In total, we identified 156 622 previously unreported variants. The allele frequencies for the majority (88.8%) were lower than 0.5% in allele frequency and predicted to be functionally deleterious. In addition, we have constructed a Japanese-specific major allele reference genome by which the number of unique mapping of the short reads in our data has increased 0.045% on average. Our results illustrate the importance of constructing an ethnicity-specific reference genome for identifying rare variants. All the collected data were centralized to a newly developed database to serve as useful resources for exploring pathogenic variations. Public access to the database is available at http://www.genome.med.kyoto-u.ac.jp/SnpDB/.

SOX9 haploinsufficiency underlies campomelic dysplasia (CD) with or without testicular dysgenesis. Current understanding of the phenotypic variability and mutation spectrum of SOX9 abnormalities remains fragmentary. Here, we report three patients with hitherto unreported SOX9 abnormalities. These patients were identified through molecular analysis of 33 patients with 46,XY disorders of sex development (DSD). Patients 1–3 manifested testicular dysgenesis or regression without CD. Patients 1 and 2 carried probable damaging mutations p.Arg394Gly and p.Arg437Cys, respectively, in the SOX9 C‐terminal domain but not in other known 46,XY DSD causative genes. These substitutions were absent from ~120,000 alleles in the exome database. These mutations retained normal transactivating activity for the Col2a1 enhancer, but showed impaired activity for the Amh promoter. Patient 3 harbored a maternally inherited ~491 kb SOX9 upstream deletion that encompassed the known 32.5 kb XY sex reversal region. Breakpoints of the deletion resided within nonrepeat sequences and were accompanied by a short‐nucleotide insertion. The results imply that testicular dysgenesis and regression without skeletal dysplasia may be rare manifestations of SOX9 abnormalities. Furthermore, our data broaden pathogenic SOX9 abnormalities to include C‐terminal missense substitutions which lead to target‐gene‐specific protein dysfunction, and enhancer‐containing upstream microdeletions mediated by nonhomologous end‐joining.

Development of resistant cultivars has been an effective method for controlling rice blast disease caused by Magnaporthe oryzae. Quantitative blast resistance genes may offer durable resistance because the selection pressure on M. oryzae to overcome resistance is low as a result of the genes’ moderate susceptibility. Because the effects of individual resistance genes are relatively small, pyramiding these genes in rice cultivars is a promising strategy. Here, we used near-isogenic and backcross lines of rice cultivar Koshihikari with single- or two-gene combinations of blast resistance genes (pi21, Pi34, and Pi35) to evaluate the suppression of leaf blast. The severity of the disease was assessed throughout the infection process. Resistance varied among the lines: Pi35 conferred the strongest resistance, while Pi34 showed the weakest effects. Two types of combined-gene interactions were observed, and they varied on the basis of gene combination and characteristic of the infection: (i) the combination of two resistance genes was more effective than either of the genes individually or (ii) the combination of two resistance genes was similar to the level of the most effective resistance gene in the pair. The most effective gene combination for the suppression of leaf blast was pi21 + Pi35.

Nucleotide binding domain and leucine-rich repeat (NLR)-containing family proteins function as intracellular immune sensors in both plants and animals. In plants, the downstream components activated by NLR family proteins and the immune response mechanisms induced by these downstream molecules are largely unknown. We have previously found that the small GTPase OsRac1, which acts as a molecular switch in rice immunity, is activated by Pit, an NLR-type resistance (R) protein to rice blast fungus, and this activation plays critical roles in Pit-mediated immunity. However, the sites and mechanisms of activation of Pit in vivo remain unknown. To clarify the mechanisms involved in the localization of Pit, we searched for consensus sequences in Pit that specify membrane localization and found a pair of potential palmitoylation sites in the N-terminal coiled-coil region. Although wild-type Pit was localized mainly to the plasma membrane, this membrane localization was compromised in a palmitoylation-deficient mutant of Pit. The palmitoylation-deficient Pit displayed significantly lower affinity for OsRac1 on the plasma membrane, thereby resulting in failures of the Pit-mediated cell death, the production of reactive oxygen species, and disease resistance to rice blast fungus. These results indicate that palmitoylation-dependent membrane localization of Pit is required for the interaction with and the activation of OsRac1 and that OsRac1 activation by Pit is vital for Pit-mediated disease resistance to rice blast fungus.

今回は歯科衛生士の将来像についての企画で「歯科 医師と歯科衛生士の連携と協働」というテーマで書か せていただく事になった. 歯科衛生士の存在なしには歯科医療は成り立たない が,ほとんどの歯科医師にとっては歯科医師の指示に 従い歯科医療の補助をする者でしかない.たしかに歯 科衛生士法 第2条には「・・歯科医師(歯科医業を なすことができる医師を含む)の直接の指導の下に歯 牙及び口腔疾患の予防処置として次に上げる行為を行 う・・・」となっている.一方チーム医療の重要性が 強調されるようになり,歯科医師とは上下の関係では なく共に歯科医療に当たる大切なパートナーであると の考え方になってきている.最近歯科衛生士の将来像 として,また歯科医師との望ましい関係とはどのよう なものかについて様々な場所で語られることとなっ た. ここでは,まず歯科衛生士の働く環境について,次 に期待される歯科衛生士像について,最後に望まれる 歯科医師と歯科衛生士の関係について述べていきた い. 以前から臨床現場での歯科衛生士の不足が叫ばれて 来た.一方歯科衛生士の就学年数が 3 年以上となり, 専門学校閉鎖,定員の削減と共に歯科衛生士の不足は ますます深刻となっている.最近では歯科医師会を中 心に離職した歯科衛生士を臨床の場に戻そうと「リカ バリー研修会」と銘打った研修会が各地で開催されて いる.筆者はこのリカバリー研修会の開催を担当した 経験があり,その際に経験した事を少し述べたい. 研修会に参加した歯科衛生士は結婚または出産で退 職した者がほとんどであった.真剣に復職を考えてい た者も多かったが家庭との両立のためほとんどがパー トでの就職を希望しており,休職後の現場復帰に対す る不安が大きくあった.また歯科衛生士が復職するに は経済的な問題が第一に挙げられる.夫のリストラや 病気などの切羽詰まった理由だけでなく,増えてくる 子供の教育費や住宅の購入費用などを捻出するために 再就職したいと考えていることが多い.収入を得るた めに復職を考えている者ほど再就職には積極的であ る.

Previously, we found that C57BL/6J (B6) mice are more prone to develop obesity than PWK mice. In addition, we analyzed reciprocal crosses between these mice and found that (PWK × B6) F1 mice, which have B6 fathers, are more likely to develop dietary obesity than (B6 × PWK) F1 mice, which have B6 mothers. These results suggested that diet-induced obesity is paternally transmitted. In this study, we performed transcriptome analysis of adipose tissues of B6, PWK, (PWK × B6) F1, and (B6 × PWK) F1 mice using next-generation sequencing. We found that paternal transmission of diet-induced obesity was correlated with genes involved in adipose tissue inflammation, metal ion transport, and cilia. Furthermore, we analyzed the imprinted genes expressed in white adipose tissue (WAT) and obesity. Expression of paternally expressed imprinted genes (PEGs) was negatively correlated with body weight, whereas expression of maternally expressed imprinted genes (MEGs) was positively correlated. In the obesity-prone B6 mice, expression of PEGs was down-regulated by a high-fat diet, suggesting that abnormally low expression of PEGs contributes to high-fat diet-induced obesity in B6 mice. In addition, using single-nucleotide polymorphisms that differ between B6 and PWK, we identified candidate imprinted genes in WAT.

Our previous studies revealed that the double variant of cytochrome P450 (CYP)105A1, R73V/R84A, has a high ability to convert vitamin D(3) to its biologically active form, 1α,25-dihydroxyvitamin D(3) [1α,25(OH)(2)D(3)], suggesting the possibility for R73V/R84A to produce 1α,25(OH)(2)D(3). Because Actinomycetes, including Streptomyces, exhibit properties that have potential advantages in the synthesis of secondary metabolites of industrial and medical importance, we examined the expression of R73V/R84A in Streptomyces lividans TK23 cells under the control of the tipA promoter. As expected, the metabolites 25-hydroxyvitamin D(3) [25(OH)D(3)] and 1α,25(OH)(2)D(3) were detected in the cell culture of the recombinant S. lividans. A large amount of 1α,25(OH)(2)D(3), the second-step metabolite of vitamin D(3), was observed, although a considerable amount of vitamin D(3) still remained in the culture. In addition, novel polar metabolites 1α,25(R),26(OH)(3)D(3) and 1α,25(S),26(OH)(3)D(3), both of which are known to have high antiproliferative activity and low calcemic activity, were observed at a ratio of 5:1. The crystal structure of the double variant with 1α,25(OH)(2)D(3) and a docking model of 1α,25(OH)(2)D(3) in its active site strongly suggest a hydrogen-bond network including the 1α-hydroxyl group, and several water molecules play an important role in the substrate-binding for 26-hydroxylation. In conclusion, we have demonstrated that R73V/R84A can catalyze hydroxylations at C25, C1 and C26 (C27) positions of vitamin D(3) to produce biologically useful compounds.

SummaryThe nucleotide-binding domain and leucine-rich repeat-containing (NLR) family proteins recognize pathogen-derived molecules and trigger immune responses in both plants and animals. In plants, the direct or indirect recognition of specific pathogen effectors by NLRs culminates in a hypersensitive response (HR) and the production of reactive oxygen species (ROS), key components of the plant defense response. However, the molecules activated by NLRs and how they induce immune responses are largely unknown. We found that the rice GTPase OsRac1 at the plasma membrane interacts directly with Pit, an NLR protein that confers resistance to the rice blast fungus. OsRac1 contributes to Pit-mediated ROS production as well as the HR and is required for Pit-mediated disease resistance in rice. Furthermore, the active form of Pit induces the activation of OsRac1 at the plasma membrane. Thus, OsRac1 is activated by Pit during pathogen attack and plays a critical role in Pit-mediated immunity in rice.

Summary The plant genome contains a large number of disease resistance (R) genes that have evolved through diverse mechanisms. Here, we report that a long terminal repeat (LTR) retrotransposon contributed to the evolution of the rice blast resistance gene Pit. Pit confers race-specific resistance against the fungal pathogen Magnaporthe grisea, and is a member of the nucleotide-binding site leucine-rich repeat (NBS-LRR) family of R genes. Compared with the non-functional allele PitNpb, the functional allele PitK59 contains four amino acid substitutions, and has the LTR retrotransposon Renovator inserted upstream. Pathogenesis assays using chimeric constructs carrying the various regions of PitK59 and PitNpb suggest that amino acid substitutions might have a potential effect in Pit resistance; more importantly, the upregulated promoter activity conferred by the Renovator sequence is essential for Pit function. Our data suggest that transposon-mediated transcriptional activation may play an important role in the refunctionalization of additional ‘sleeping’R genes in the plant genome.

Cystic lymphangiomas are congenital malformations of the lymphatic system, such as benign tumors that may be seen in the neck of infants. Surgical therapy entails risk of complications, depending on the location and extent of the tumor. Therefore, alternative treatments such as sclerotherapy with OK-432, have been developed. We report here a case of cystic lymphangioma treated by ultrasonography (US)-guided percutaneous aspiration and intraregional injection of OK-432 under general anesthesia. The difficulty in this case was the location of the tumor, which contacted the trachea. If OK-432 were to induce an inflammatory response in the trachea, it could lead to tracheal stenosis and respiratory disturbances. Therefore, this case required endotracheal Intubation and mechanical ventilation after injection therapy for three days. OK-432 sclerotherapy is an effective treatment for cystic lymphangiomas in infants, and should be considered before surgical excision.

Sepsis is characterized by various symptoms, signs and underlying pathophysiology. To investigate possible mechanisms underlying this diversity, we compared the cardiovascular effects of lipopolysaccharide (LPS) derived from Escherichia coli (E-LPS) with those of LPS from Pseudomonas aeruginosa (P-LPS) in rats. We also examined the possible roles of tumor necrosis factor-alpha (TNF-alpha) and oxidative stress in LPS-induced cardiovascular damage. E-LPS (10 mg/kg body weight) or P-LPS (2 mg/kg body weight) was administered intravenously to Wistar rats. Echocardiography was serially performed. E-LPS induced an increase in left ventricular fractional shortening that persisted for at least 6 h, whereas P-LPS elicited an initial increase and a subsequent decrease in this parameter. Histological analysis revealed that P-LPS induced interstitial edema, congestion, intramyocardial bleeding, myocardial necrosis, infiltration of inflammatory cells, and formation of fibrin thrombi in the heart, whereas no pathological changes were apparent in the hearts of rats treated with E-LPS. Furthermore, the plasma concentration of TNF-alpha in rats treated with P-LPS was greater than that in rats treated with E-LPS, but the glutathione redox ratio in the heart was not affected by either type of LPS. In conclusion, E-LPS and P-LPS induced distinct patterns of functional and structural responses in the cardiovascular systems of rats. These differential responses may be attributable in part to the difference in the associated increases in the plasma concentration of TNF-alpha. The cardiovascular effects of LPS thus depend on the causative organisms.

The hypersensitive response (HR) of plants is one of the earliest responses to prevent pathogen invasion. A brown dot lesion on a leaf is visual evidence of the HR against the blast fungus Magnaporthe oryzae in rice, but tracking the browning process has been difficult. In this study, we induced the HR in rice cultivars harboring the blast resistance gene Pit by inoculation of an incompatible M. oryzae strain, which generated a unique resistance lesion with a brown ring (halo) around the brown fungal penetration site. Inoculation analysis using a plant harboring Pit but lacking an enzyme that catalyzes tryptamine to serotonin showed that high accumulation of the oxidized form of serotonin was the cause of the browning at the halo and penetration site. Our analysis of the halo browning process in the rice leaf revealed that abscisic acid enhanced biosynthesis of serotonin under light conditions, and serotonin changed to the oxidized form via hydrogen peroxide produced by light. The dramatic increase in serotonin, which has a high antioxidant activity, suppressed leaf damage outside the halo, blocked expansion of the browning area and attenuated inhibition of plant growth. These results suggest that serotonin helps to reduce biotic stress in the plant by acting as a scavenger of oxygen radicals to protect uninfected tissues from oxidative damage caused by the HR. The deposition of its oxide at the HR lesion is observed as lesion browning.

連続的携行式腹膜透析(以下PD)は自己管理を主体とした在宅の透析療法である。今回バッグ交換困難な手指変形を有する59歳女性の関節リウマチ患者に対して，SMAP法を用いたPD導入を行ない自己管理可能となった事例を経験した。事例は著明な手指関節変形を有する末期腎不全患者でUV-Fツインバッグシステムを用いた自己管理によるPDを希望した。バッグ交換は1日4回行ない1回のバッグ交換には数種類のクランプ操作を必要とするが，手指関節変形により通常のバッグ交換操作は不可能であった。既存のデバイスの活用と新たなデバイスを作製することにより自己管理によるバッグ交換が可能となった。SMAP法はPD開始までに時間的猶予があるため自己管理方法の検討と操作習得の練習期間の確保が可能であり，手指関節変形を有する関節リウマチ患者に対してSMAP法でのPD導入は有用であった。

いもち病真性抵抗性遺伝子Pizに連鎖する5種類の1塩基多型（SNP）マーカーの有効性を検証するために， Pizを持たないコシヒカリとPizを有する新潟早生とのF1にコシヒカリを連続戻し交配した集団において， Pizが座乗する染色体上のゲノム領域が新潟早生型でその両側がコシヒカリ型に組み換わっている個体を選抜するとともに，かついもち病真性抵抗性を有することを確認し，SNPマーカーの有効性を示した．

Our previous study indicated that the acinar cells regenerated actively in the rat pancreatic remnant after partial (90%) pancreatectomy. On that occasion, a twofold higher mitotic activity was observed in the cells of paraduodenal than those of the medial and stump regions (regional difference). Since cholecystokinin (CCK) has been known to have various physiological functions in the digestive system, especially in the pancreas, through the CCK-A receptor, we determined the effect of a CCK-A receptor competitive antagonist (CCK-RA) (SR27897B) on pancreatic regeneration after pancreatectomy. A group of Wistar rats was administered 1 mg/kg CCK-RA twice daily for 5 days. On the 2nd day of administration, partial pancreatectomy was performed. Proliferation activity of both exocrine and endocrine cells was found to be enhanced uniformly 5 days after the pancreatectomy, and the effect was inhibited by CCK-RA. Plasma CCK level was increased by 90% pancreatectomy and decreased by CCK-RA. CCK was hypothesized to be an important factor of pancreatic exocrine and endocrine regeneration. Administration with CCK-RA suppressed mitotic activity uniformly in the three regions of paraduodenal, medial and stump. From the evidence, it was presumed that regional difference in regeneration was produced not by paracrine factor from the duodenum but by some other unknown mechanism including anatomical difference in vascular and autonomic nervous system.

Summary. T-cell granular lymphocyte-proliferative disorder (T-GLPD) is characterized by the proliferation of cytotoxic T lymphocytes, and is often associated with pure red cell aplasia (PRCA). The mechanism involved in the development of PRCA in T-GLPD is unknown. Peripheral blood mononuclear cells were isolated from 20 patients with T-GLPD. Ten patients had associated PRCA. Granular lymphocytes (GLs) of T-GLPD are positive for CD94, but not NKG2A. To clarify the functional role of CD94 in T-GLPD, we performed a cytotoxicity assay against the trophoblast cell line, BeWo, which is known to express human leucocyte antigen (HLA)-E, a natural ligand of CD94, and is deficient in other HLA class I and class II antigens. GLs isolated from T-GLPD with PRCA patients killed BeWo cells more significantly than GLs from T-GLPD without PRCA patients. Furthermore, GLs from T-GLPD with PRCA were significantly stimulated by a monoclonal antibody against CD94, whereas those of T-GLPD without PRCA were not. Taken together, HLA-E, a ligand of CD94, was suggested to stimulate CD94+ cells to kill HLA-E+ cells in T-GLPD with PRCA. GLs of T-GLPD with PRCA have a potential positive activity against HLA-E+ cells, whereas GLs from T-GLPD without PRCA do not. CD94 may play a key role in the pathogenesis of PRCA in T-GLPD.

We treated rapidly growing Jurkat cells with 40 nmol/l of doxorubicin for 72 h. After 36 h, the G2-arrested cells became larger and some of them started endoreplication. Nuclear staining with Hoechst 33342 combined with propidium iodide (PI) exclusion revealed that about 90% of the cells were necrotic at 72 h, although apoptotic cells accounted for only 8%. Incubation with 40 nmol/l of aclarubicin or cytosine beta-d-arabinofuranoside for 60 h induced necrosis both in Jurkat and ml-1 cells. Pre-necrotic Jurkat cells incubated with 40 nmol/l of doxorubicin had much higher intracellular reactive oxygen species (ROS) levels than pre-apoptotic ones. Addition of Tempol or Desferal accelerated doxorubicin-induced necrosis and partially converted it into apoptosis. Both antioxidants reduced surviving colony numbers of prenecrotic Jurkat cells. n-acetyl-l-cysteine had little effect on the apoptotic conversion but profoundly accelerated necrosis. Because an apoptosis-resistant Jurkat subclone was also refractory to doxorubicin-induced necrosis, apoptosis and necrosis might share some common pathways. Low-dose doxorubicin increased micronuclei-positive cell percentages and also suppressed high-dose doxorubicin-induced apoptosis in Jurkat and ml-1 cells. Some of the prenecrotic cells, therefore, might survive and obtain genomic instability. Antioxidants may be useful to suppress, at least to some extent, this vicious consequence.

Sterols have important functions in membranes and signaling. Plant sterols are synthesized via the isoprenoid pathway by cyclization of 2,3-oxidosqualene to cycloartenol. Plants also convert 2,3-oxidosqualene to other sterol-like cyclization products, including the simple triterpene β-amyrin. The function of β-amyrin per se is unknown, but this molecule can serve as an intermediate in the synthesis of more complex triterpene glycosides associated with plant defense. β-Amyrin is present at low levels in the roots of diploid oat (Avena strigosa). Oat roots also synthesize the β-amyrin–derived triterpene glycoside avenacin A-1, which provides protection against soil-borne diseases. The genes for the early steps in avenacin A-1 synthesis [saponin-deficient 1 and 2 (Sad1 and Sad2)] have been recruited from the sterol pathway by gene duplication and neofunctionalization. Here we show that Sad1 and Sad2 are regulated by an ancient root developmental process that is conserved across diverse species. Sad1 promoter activity is dependent on an L1 box motif, implicating sterol/lipid-binding class IV homeodomain leucine zipper transcription factors as potential regulators. The metabolism of β-amyrin is blocked in sad2 mutants, which therefore accumulate abnormally high levels of this triterpene. The accumulation of elevated levels of β-amyrin in these mutants triggers a “superhairy” root phenotype. Importantly, this effect is manifested very early in the establishment of the root epidermis, causing a greater proportion of epidermal cells to be specified as root hair cells rather than nonhair cells. Together these findings suggest that simple triterpenes may have widespread and as yet largely unrecognized functions in plant growth and development.

Background Rice blast is a destructive disease caused by Magnaporthe oryzae, and it has a large impact on rice production worldwide. Compared with leaf blast resistance, our understanding of panicle blast resistance is limited, with only one panicle blast resistance gene, Pb1, isolated so far. The japonica cultivar Miyazakimochi shows resistance to panicle blast, yet the genetic components accounting for this resistance remain to be determined. Results In this study, we evaluated the panicle blast resistance of populations derived from a cross between Miyazakimochi and the Bikei 22 cultivar, which is susceptible to both leaf and panicle blast. The phenotypic analyses revealed no correlation between panicle blast resistance and leaf blast resistance. Quantitative trait locus (QTL) analysis of 158 recombinant inbred lines using 112 developed genome-wide and 35 previously reported polymerase chain reaction (PCR) markers revealed the presence of two QTLs conferring panicle blast resistance in Miyazakimochi: a major QTL, qPbm11, on chromosome 11; and a minor QTL, qPbm9, on chromosome 9. To clarify the contribution of these QTLs to panicle blast resistance, 24 lines homozygous for each QTL were selected from 2,818 progeny of a BC2F7 backcrossed population, and characterized for disease phenotypes. The panicle blast resistance of the lines harboring qPbm11 was very similar to the resistant donor parental cultivar Miyazakimochi, whereas the contribution of qPbm9 to the resistance was small. Genotyping of the BC2F7 individuals highlighted the overlap between the qPbm11 region and a locus of the panicle blast resistance gene, Pb1. Reverse transcriptase PCR analysis revealed that the Pb1 transcript was absent in the panicles of Miyazakimochi, demonstrating that qPbm11 is a novel genetic component of panicle blast resistance. Conclusions This study revealed that Miyazakimochi harbors a novel panicle blast resistance controlled mainly by the major QTL qPbm11. qPbm11 is distinct from Pb1 and could be a genetic source for breeding panicle blast resistance, and will improve understanding of the molecular basis of host resistance to panicle blast.

A fusion between the plastid psbA promoter and the green fluorescent protein gene (gfp) was introduced into the tobacco chloroplast genome by stable plastid transformation. GFP was synthesized actively and exclusively in the chloroplasts. Tubular projections filled with GFP but containing no chlorophyll were visualized for the first time in chloroplasts of these transplastomic plants. Occasionally, the tubules connect chloroplasts with each other, suggesting the possibility of the exchange of endogenous proteins. However, the fusion of protoplasts between the transplastomic and wild-type plants showed that such chloroplast connections might be rare in mesophyll protoplasts.

The relationship between the index of general trust and decision making process of civil engineers is examined by using analysis of answers to question sheet. From the analysis, it appears that there is a close relationships between them. As the score of general trust increases, the decision of engineers becomes more positive. It is also found out that socialization process of civil engineers is completed at around 30 years old. It is necessary to do ethics education before this age.

We have cloned the gene encoding RNase HII (RNase HII Pk ) from the hyperthermophilic archaeon Pyrococcus kodakaraensis KOD1 by screening of a library for clones that suppressed the temperature-sensitive growth phenotype of an rnh mutant strain of Escherichia coli . This gene was expressed in an rnh mutant strain of E. coli , the recombinant enzyme was purified, and its biochemical properties were compared with those of E. coli RNases HI and HII. RNase HII Pk is composed of 228 amino acid residues (molecular weight, 25,799) and acts as a monomer. Its amino acid sequence showed little similarity to those of enzymes that are members of the RNase HI family of proteins but showed 40, 31, and 25% identities to those of Methanococcus jannaschii , Saccharomyces cerevisiae , and E. coli RNase HII proteins, respectively. The enzymatic activity was determined at 30°C and pH 8.0 by use of an M13 DNA-RNA hybrid as a substrate. Under these conditions, the most preferred metal ions were Co 2+ for RNase HII Pk , Mn 2+ for E. coli RNase HII, and Mg 2+ for E. coli RNase HI. The specific activity of RNase HII Pk determined in the presence of the most preferred metal ion was 6.8-fold higher than that of E. coli RNase HII and 4.5-fold lower than that of E. coli RNase HI. Like E. coli RNase HI, RNase HII Pk and E. coli RNase HII cleave the RNA strand of an RNA-DNA hybrid endonucleolytically at the P-O3′ bond. In addition, these enzymes cleave oligomeric substrates in a similar manner. These results suggest that RNase HII Pk and E. coli RNases HI and HII are structurally and functionally related to one another.

Background/Aims: Missense, nonsense, and splice mutations in the Fibroblast Growth Factor 8(FGF8) have recently been identified in patients with hypothalamo-pituitary dysfunction and craniofacial anomalies. Here, we report a male patient with a frameshift mutation in FGF8. Case Report: The patient exhibited micropenis, craniofacial anomalies, and ventricular septal defect at birth. Clinical evaluation at 16 years and 8 months of age revealed delayed puberty, hyposmia, borderline mental retardation, and mild hearing difficulty. Endocrine findings included gonadotropin deficiency and primary hypothyroidism. Results: Molecular analysis identified a de novo heterozygous p.S192fsX204 mutation in the last exon of FGF8. RT-PCR analysis of normal human tissues detected FGF8 expression in the genital skin, and whole-mount in situ hybridization analysis of mouse embryos revealed Fgf8 expression in the anlage of the penis. Conclusion: The results indicate that frameshift mutations in FGF8 account for a part of the etiology of hypothalamo-pituitary dysfunction. Micropenis in patients with FGF8 abnormalities appears to be caused by gonadotropin deficiency and defective outgrowth of the anlage of the penis.

The western flower thrips (WFT), Frankliniella occidentalis, was first discovered in a southern area of Fukuoka Prefecture in April, 1994. Our census surveys revealed that the distribution of WFT has been limited to the southern part and a few restricted areas in Fukuoka Prefecture even one and a half years after its' initial discovery. In strawberry greenhouses, the number of WFT adults captured on blue sticky traps remained low during winter. However, the number increased rapidly in late March and early April. In a rose greenhouse, a similar tendency in the number of WFT was observed. Judging from the abundance pattern in greenhouses and the low temperature conditions outside the greenhouses, the WFT populations overwintering in greenhouses appeared to be a major source of the spring populations. From the control perspective, applications of insecticides immediately after planting and in early spring were thought to be crucial. The number of WFT adults captured in traps placed around a strawberry greenhouse and its surrounding vegetation showed that WFT adults dispersed from the greenhouse during late April and early May, and then colonized the surrounding vegetation. Thus, WFT appeared to utilize greenhouse crops and their surrounding vegetation successively as temporal habitats.

PURPOSE: To evaluate the cross-sectional configurations of peripapillary atrophy (PPA)-alpha and -beta in ophthalmologically normal subjects using spectral domain-optical coherence tomography (SD-OCT). METHODS: One hundred twenty normal subjects had a complete ophthalmic examination including axial length measurement, standard automated perimetry, fundus imaging with photography, and SD-OCT (3D OCT-1000; Topcon Inc., Tokyo, Japan). PPA-alpha and -beta were identified in color photographs of the optic disc. Cross-sectional B-mode images of the peripapillary retina and sclera, including PPA-alpha and -beta obtained with SD-OCT, were analyzed. RESULTS: Of 120 normal eyes, 120 (100%) had PPA-alpha and 90 (75%) had PPA-beta. In OCT images of the peripapillary retina, the ganglion cell layer and the inner and outer plexiform layers were observed to end in a tapering fashion at the edge of the optic disc, whereas the retinal nerve fiber layer continued into the optic cup. The external limiting membrane (ELM), inner-outer segments (IS-OS), and retinal pigment epithelium(RPE)/Bruch's membrane complex were significantly more commonly absent before the optic disc edge within the PPA-beta compared with areas outside the PPA-beta (P < 0.0001). Specific findings in the peripapillary area including slope and step configurations of the scleral bed and hump- and wedge-shaped appearances of the RPE-Bruch's membrane complex were identified in 63 (52.5%), 6 (5.0%), 19 (15.8%), and 6 (5.0%) of 120 eyes, respectively. The presence of the step configuration was associated with myopia and longer axial length (P = 0.0014 and 0.0105, respectively). CONCLUSIONS: The cross-sectional anatomic configurations of the peripapillary atrophy were evaluated by using SD-OCT. The termination of the retinal layers and configurations of the scleral bed in the peripapillary area varied among normal subjects.

The 14-epimer of MART-10, namely 14-epi-MART-10 (14-epi-2alpha-(3-hydroxypropyl)-1alpha,25-dihydroxy-19-norvitamin D3) and its 2-epimeric analog (14-epi-MART-11) were efficiently synthesized using the Julia coupling reaction to connect between the C5 and C6 positions (steroid numbering). An A-ring precursor was prepared from (-)-quinic acid as shown in the previous MART-10 synthesis. The novel 14-epi-CD-ring coupling partner with an elongated two carbon unit as a sulfone was synthesized from 14-epi-25-hydroxy Grundmann's ketone in good yield. The subsequent coupling reaction followed by a deprotection step afforded a mixture of 14-epi-MART-10 and 14-epi-MART-11 in 40% yield. To separate 14-epi-MART-10 and 14-epi-MART-11, each primary hydroxyl group was esterified with a pivaloyl group and the resulting pivalates 2alpha and 2beta were separated by high performance liquid chromatography. After the separation, the C2-stereochemistry of each (2alpha or 2beta) was determined by 1H NMR (nuclear magnetic resonance) studies including NOE (nuclear Overhauser effect) experiments. The pivaloyl group was removed under basic conditions to obtain the target molecules of 14-epi-MART-10 and 14-epi-MART-11, respectively. The VDR (vitamin D receptor)-binding affinity, HL-60 (human promyelocytic leukemia) cell differentiation activity, antiproliferative activity in PZ-HPV-7 (immortalized normal prostate) cells and transactivation activity of the osteocalcin promoter in HOS (human osteoblast cell line) cells (serum-free conditions) were investigated. In addition, the effects on bone mineral density (BMD) and the blood and urine calcium concentrations of ovariectomized (OVX) rats were examined. 14-epi-MART-10 has much greater antiproliferative and cell differentiation activities compared to 1alpha,25-dihydroxyvitamin D3 (1alpha,25(OH)2D3).

持続性低血圧透析患者の臨床的特徴と看護について検討するために, 透析前収縮期血圧100mmHg以下の低血圧者10名A群と正常血圧者11名B群とについて比較した.両群の透析歴はA群11.3±6年, B群10.4±4年であり, 透析前平均血圧はA群69±7mmHg, B群では99.6±8mmHgであった.A群は合併症に2次性副甲状腺機能亢進症および手根管症候群を多数認めたが, 両群において副甲状腺ホルモン, β2-マイクログロブリン値には有意差を認めなかった, また貧血との相関関係も認められなかった. 心胸比, 体重増加率, 摂取栄養量においても両群間に差を認めなかった. A群の10例中8例においては, 循環血液量減少予防または低血圧の治療が必要であった. 血圧下降症状出現時の血圧値は各々の症例でほぼ一定であり, 透析中の血圧維持レベルを決めることにより下降防止が可能であった. また水分, 塩分摂取量の管理により適正体重を維持することが重要と考えられた.

Aims Reduction of left atrial appendage (LAA) flow velocity (FV) is a risk factor for thrombus formation and increases the risk of stroke in patients with atrial fibrillation (AF). Furthermore, LAA morphology is correlated with stroke in patients with AF. The aim of this study was to correlate LAAFV with LAA morphology in patients with AF. Methods and results We studied 96 patients (age 59.0 ± 10.2 years, 75% male) referred for radiofrequency catheter ablation for paroxysmal AF. All patients underwent computed tomography (CT) and transthoracic and transoesophageal echocardiography during sinus rhythm. LAA morphology was classified as one of the four types (chicken wing, windsock, cactus, and cauliflower) on CT images. There were significant differences in LAAFV among LAA morphologies (chicken wing 73.7 ± 21.9 cm/s, windsock 61.9 ± 19.6 cm/s, cactus 55.3 ± 14.1 cm/s, cauliflower 52.7 ± 18.1 cm/s, P = 0.008). Post hoc multiple comparisons showed that LAAFV was higher in patients with chicken wing than in those with cactus ( P = 0.006, vs . chicken wing) and cauliflower ( P = 0.006, vs . chicken wing), but not with windsock ( P = 0.102). After adjustment for clinical and LAA anatomical covariates (orifice area, volume, and trabeculation), multiple linear regression analyses revealed that LAA morphology was an independent determinant of LAAFV [chickens wing: standardized partial regression coefficients ( β ) = 0.317, P = 0.0014; windsock: β = 0.303, P = 0.038]. Conclusion LAA morphology is a significant determinant of LAAFV, suggesting an underlying mechanism for the association between LAA morphology and embolic events.

The biosynthetic gene cluster of goadsporin, a polypeptide antibiotic containing thiazole and oxazole rings, was cloned from Streptomyces sp. TP-A0584. The cluster contains a structural gene, godA, and nine god (goadsporin) genes involved in post-translational modification, immunity and transcriptional regulation. Although the gene organization is similar to typical bacteriocin biosynthetic gene clusters, each goadsporin biosynthetic gene shows low homology to these genes. Goadsporin biosynthesis is initiated by the translation of godA, and the subsequent cyclization, dehydration and acetylation are probably catalysed by godD, godE, godF, godG and godH gene products. godI shows high similarity to the 54 kDa subunit of the signal recognition particle and plays an important role in goadsporin immunity. Furthermore, four goadsporin analogues were produced by site-directed mutagenesis of godA, suggesting that this biosynthesis machinery is used for the heterocyclization of peptides.

The effect of laughter therapy on the plasma levels of renin, angiotensinogen, and prorenin was investigated in patients with type 2 diabetes. In the diabetic patients, the mean plasma renin concentrations were 24.6+/-12.1 ng/ml/h in the first observation (at the beginning of laughter therapy), 8.2+/-3.4 ng/ml/h in the second observation (three months after the beginning of laughter therapy) and 7.7+/-1.7 ng/ml/h in the third observation (six months after the beginning of laughter therapy). The mean plasma angiotensinogen concentrations in the 1st, 2nd and 3rd observations were 0.19+/-0.08, 0.47+/-0.12, 0.42+/-0.14 microg/ml, respectively. The mean plasma prorenin concentrations in the 1st, 2nd and 3rd observations during the laughter therapy were 195.1+/-66.2, 193.4+/-88.2 and 170.7+/-52.5 pg/ml, respectively. Plasma renin concentrations were significantly decreased (p

Most plastid promoters recognized by bacteria-like plastid RNA polymerase (PEP) are similar to E. coli sigma(70)-type promoters comprising "-35" and "-10" elements. Among them, psbA promoter is unique in bearing additional elements between the conserved -35 and -10 elements. The psbA promoter activity is differentially maintained in the mature chloroplasts where the activity of most PEP promoters declines. Previously, we identified two types of PEP activities in wheat seedlings [Satoh et al. (1999) Plant J. 18: 407]; PEP present in the mature chloroplasts of the leaf tip (tip-type PEP) can initiate transcription from the -35-destructed psbA promoter, but the -35 element is essential for transcription by PEP present in immature chloroplasts of the leaf base (base-type PEP). To reveal which type of PEP functions in various types of plastids in tobacco, we analyzed the tobacco psbA promoter by means of a transplastomic approach. The promoter core context (-42 to +9) was sufficient for developmental regulation of the psbA promoter activity. The -35 promoter element was important for transcription initiation at the psbA promoter in all types of plastids, including chloroplasts in mature leaves, leucoplasts in roots, etioplasts in etiolated cotyledons. The conclusion is that the PEP bearing a promoter preference, similar to the wheat base-type PEP, functions dominantly in tobacco chloroplasts.