Vegetative Propagation of Yakon by Shoot Apex, Node and Callus Cultures

ヤーコンの無病苗の増殖のために,茎頂培養,節培養,カルス培養を試みた.茎頂培養は,0.01mg/l NAAとBAを添加したMS培地に0.3～0.5mmの茎頂を植え付けて約1か月間培養することにより,植物体となった.これはバーミキュライトで順化することが可能であった. 節培養は,葉を切り捨てた無菌植物の節を0.01mg/l NAA及びBA添加又は無添加のMS又はハイポネックス培地に植え付けることにより,節当たり約2苗条得ることができ,そのままの培地で培養することにより発根した.基部側の下半分の節からの苗条の生育が非常に良く,MS,ハイポネックスいずれの培地でもホルモン無添加培地でよく生育した. カルス培養は,供試材料として無菌植物の葉,茎,根を比較したところ茎で一番大きなカルスができたが,培地によっては再現性にかける.茎のみならず葉でも根でも0.lmg/l 2,4-DとBAを添加したMS培地で大きなカルスが出来た. 茎頂培養で形成された植物の茎頂を,0.1mg/l 2,4-Dと1mg/l BAを添加した培地に移植してカルスを形成させ,さらに0.1mg/l BA添加培地に移植したところ,低率ながら不定胚が形成された.これは植物体にまで生長した

Effects of Media on Overcoming Vitrification of Carnation in Apex Culture

スプレータイプのカーネーション“ナティラ”と“ピンクカジノ”を供試し,茎頂培養に適した培地の支持体を検討した.基本培地としてハイポネックス,NH4NO3,MgSO4・7H20,Na-EDTA,塩酸ピリドキシン,カイネチン,NAA,しょ糖を含む培地に,支持体として寒天0.6% ,gelrite0.2% またはペーパーブリッジを別々に添加した.茎頂は0.3～0.5mmの部分を切り出し,培地に植え付けた.3か月培養の結果,寒天培地上で最も高率に正常な植物が生長し,gelrite上では最も水浸状の,しかも多芽体が多かった.生体重と乾燥重の比較では,gelrite,寒天,ペーパーブリッジの順であったが,乾物率は寒天が最も高く,gelriteは低かった

A Serine Palmitoyltransferase Inhibitor Blocks Hepatitis C Virus Replication in Human Hepatocytes

Background & AimsHost cell lipid rafts form a scaffold required for replication of hepatitis C virus (HCV). Serine palmitoyltransferases (SPTs) produce sphingolipids, which are essential components of the lipid rafts that associate with HCV nonstructural proteins. Prevention of the de novo synthesis of sphingolipids by an SPT inhibitor disrupts the HCV replication complex and thereby inhibits HCV replication. We investigated the ability of the SPT inhibitor NA808 to prevent HCV replication in cells and mice.MethodsWe tested the ability of NA808 to inhibit SPT’s enzymatic activity in FLR3-1 replicon cells. We used a replicon system to select for HCV variants that became resistant to NA808 at concentrations 4- to 6-fold the 50% inhibitory concentration, after 14 rounds of cell passage. We assessed the ability of NA808 or telaprevir to inhibit replication of HCV genotypes 1a, 1b, 2a, 3a, and 4a in mice with humanized livers (transplanted with human hepatocytes). NA808 was injected intravenously, with or without pegylated interferon alfa-2a and HCV polymerase and/or protease inhibitors.ResultsNA808 prevented HCV replication via noncompetitive inhibition of SPT; no resistance mutations developed. NA808 prevented replication of all HCV genotypes tested in mice with humanized livers. Intravenous NA808 significantly reduced viral load in the mice and had synergistic effects with pegylated interferon alfa-2a and HCV polymerase and protease inhibitors.ConclusionsThe SPT inhibitor NA808 prevents replication of HCV genotypes 1a, 1b, 2a, 3a, and 4a in cultured hepatocytes and in mice with humanized livers. It might be developed for treatment of HCV infection or used in combination with pegylated interferon alfa-2a or HCV polymerase or protease inhibitors

Role of Serine Racemase in Behavioral Sensitization in Mice after Repeated Administration of Methamphetamine

BACKGROUND: The N-methyl-D-aspartate (NMDA) receptors play a role in behavioral abnormalities observed after administration of the psychostimulant, methamphetamine (METH). Serine racemase (SRR) is an enzyme which synthesizes D-serine, an endogenous co-agonist of NMDA receptors. Using Srr knock-out (KO) mice, we investigated the role of SRR on METH-induced behavioral abnormalities in mice. METHODOLOGY/PRINCIPAL FINDINGS: Evaluations of behavior in acute hyperlocomotion, behavioral sensitization, and conditioned place preference (CPP) were performed. The role of SRR on the release of dopamine (DA) in the nucleus accumbens after administration of METH was examined using in vivo microdialysis technique. Additionally, phosphorylation levels of ERK1/2 proteins in the striatum, frontal cortex and hippocampus were examined using Western blot analysis. Acute hyperlocomotion after a single administration of METH (3 mg/kg) was comparable between wild-type (WT) and Srr-KO mice. However, repeated administration of METH (3 mg/kg/day, once daily for 5 days) resulted in behavioral sensitization in WT, but not Srr-KO mice. Pretreatment with D-serine (900 mg/kg, 30 min prior to each METH treatment) did not affect the development of behavioral sensitization after repeated METH administration. In the CPP paradigm, METH-induced rewarding effects were demonstrable in both WT and Srr-KO mice. In vivo microdialysis study showed that METH (1 mg/kg)-induced DA release in the nucleus accumbens of Srr-KO mice previously treated with METH was significantly lower than that of the WT mice previously treated with METH. Interestingly, a single administration of METH (3 mg/kg) significantly increased the phosphorylation status of ERK1/2 in the striatum of WT, but not Srr-KO mice. CONCLUSIONS/SIGNIFICANCE: These findings suggest first, that SRR plays a role in the development of behavioral sensitization in mice after repeated administration of METH, and second that phosphorylation of ERK1/2 by METH may contribute to the development of this sensitization as seen in WT but not Srr-KO mice

Criterion and Construct Validity of the CogState Schizophrenia Battery in Japanese Patients with Schizophrenia

BACKGROUND: The CogState Schizophrenia Battery (CSB), a computerized cognitive battery, covers all the same cognitive domains as the Measurement and Treatment Research to Improve Cognition in Schizophrenia (MATRICS) Consensus Cognitive Battery but is briefer to conduct. The aim of the present study was to evaluate the criterion and construct validity of the Japanese language version of the CSB (CSB-J) in Japanese patients with schizophrenia. METHODOLOGY/PRINCIPAL FINDINGS: Forty Japanese patients with schizophrenia and 40 Japanese healthy controls with matching age, gender, and premorbid intelligence quotient were enrolled. The CSB-J and the Brief Assessment of Cognition in Schizophrenia, Japanese-language version (BACS-J) were performed once. The structure of the CSB-J was also evaluated by a factor analysis. Similar to the BACS-J, the CSB-J was sensitive to cognitive impairment in Japanese patients with schizophrenia. Furthermore, there was a significant positive correlation between the CSB-J composite score and the BACS-J composite score. A factor analysis showed a three-factor model consisting of memory, speed, and social cognition factors. CONCLUSIONS/SIGNIFICANCE: This study suggests that the CSB-J is a useful and rapid automatically administered computerized battery for assessing broad cognitive domains in Japanese patients with schizophrenia

Background Coloration of Squamous Epithelium in Esophago-Pharyngeal Squamous Cell Carcinoma: What Causes the Color Change?

Objectives: This study aims to clarify the cause of background coloration in the epithelia between each dilated intra papillary capillary loop in esophago-pharyngeal squamous cell carcinoma. Design: This is a single center retrospective study including 124 patients with 160 lesions who underwent esophagogastroduodenoscopy in Nagasaki University Hospital from September 2007 to March 2012; a detailed comparison between endoscopic images and pathology was performed. Immunohistological assessment using anti-human hemoglobin antibody (anti-Hb Ab) was performed to verify the presence of hemoglobin (Hb) component in the cancer cells. Real-time polymerase chain reaction (RT-PCR) and in situ hybridization (ISH) on Hb-β mRNA were performed to assess the production of Hb component within the cancer cells. Results: A strong positivity for anti-Hb Ab was observed in the squamous cell carcinoma area, whereas non-cancerous mucosa showed no immunopositivity for Hb. The concordance rate between anti-Hb Ab immunoreactivity and the presence of BC was as high as 80.9%. The amount of Hb-β mRNA expression was three times higher in cancer tissues compared with the surrounding non-cancerous mucosa. ISH images showed that the expression exclusively occurred in cancer cells, indicating that Hb is probably produced within cancer cells. Conclusions: The background coloration observed is partly due to an extravascular component of Hb. RT-PCR and ISH analyses indicate that Hb is produced within cancer cells

Finishing the euchromatic sequence of the human genome

The sequence of the human genome encodes the genetic instructions for human physiology, as well as rich information about human evolution. In 2001, the International Human Genome Sequencing Consortium reported a draft sequence of the euchromatic portion of the human genome. Since then, the international collaboration has worked to convert this draft into a genome sequence with high accuracy and nearly complete coverage. Here, we report the result of this finishing process. The current genome sequence (Build 35) contains 2.85 billion nucleotides interrupted by only 341 gaps. It covers ∼99% of the euchromatic genome and is accurate to an error rate of ∼1 event per 100,000 bases. Many of the remaining euchromatic gaps are associated with segmental duplications and will require focused work with new methods. The near-complete sequence, the first for a vertebrate, greatly improves the precision of biological analyses of the human genome including studies of gene number, birth and death. Notably, the human enome seems to encode only 20,000-25,000 protein-coding genes. The genome sequence reported here should serve as a firm foundation for biomedical research in the decades ahead

Multiplication of Chinese Yam 'Ichouimo' (Dioscorea batatus Decne.)by Multiple Buds Culture

ヤマノイモ類は栄養繁殖されるのでウイルス罹病株が多く,かつ繁殖率が低いので苗薯が高価である. そのため無病のイチョウイモ(Dioscorea batatus Decne)の多芽体培養による増殖を試みた. Murashige&Skoog培地を基本培地とし,種々の支持体や植物ホルモンを添加し,ショ糖濃度も変えた. 培養条件は,25℃,20001x人工光による16時間日長とした. 多芽体形成のために,栽培植物のムカゴを無菌培養し,発芽してきた小植物の,1腋芽を付けた茎切片を外植体とした. 20mg/lアンシミドール添加基本培地に植え付けたところ,多芽体が形成された. この多芽体を切り分け,アンシミドール0又は10mg/l添加に植え付け,40日間振盪又は回転培養を行ったとのろ,いずれの方法でも多芽体が形成され,特にアンシミドール添加培地の振盪培養区で増殖率が高く,無添加培地では苗条が伸長した. この多芽体を切り分け,10mg/lアンシミドール,2% ショ糖添加液体培地で振盪培養し,46日間隔で継代培養を7回繰り返した. 増殖率,形成された多芽体の大きさに世代毎の大きな差は見られず,増殖率はほぼ30倍前後であった. これらの多芽体を苗化するため,切り分けた外植体を寒天0.3または0.7% ,gelrite 0.1または0.2% 添加基本培地のいずれかに植え付け,10,20,30日間培養後,順化した. いずれの区でも順化したが,培養期間が長い程発根が早く,gelrite培地の方が短期間の培養でも早く発根した. 多芽体を切り分け,0.2% gelriteと3% ショ糖添加基本培地を入れた培養瓶に植え付け室内に放置しておいた処,2～3か月後にミニチューバーが出来,冬季に一度地上部が枯れた後,春になって既に出来ていたミニチューバーが発芽し,それからの苗条が伸長し,その基部や節にまたミニチューバーが形成された. 以上イチョウイモでは多芽体培養により,1年で307本の小植物を増殖出来,さらに1芽より1年で約1個のミニチューバーが出来る事が分かった