The Double Polymerase Chain Reaction with Consensus Primers Permits Rapid and Sensitive Detection of Genital Human Papillomavirus Oncogenes

We have developed a sensitive procedure for the detection of relatively low copy numbers of multiple genital human papillomaviruses (HPVs) using the polymerase chain reaction (PCR). HPV DNAs were detected by agarose gel electrophoresis and ethidium bromide staining after 2 rounds of PCR amplification (double PCR) with outer and inner consensus primer pairs for HPV-6, 11, 16, 18, 31, 33, 52, and 58. The detection limit of this method (i. e., 10?? copy of HPV DNA per cell in 1 μg cell DNA) was sufficient for analysis of cervical intraepithelial neoplasia (CIN) specimens. Overall prevalence rate of HPV was 100% in 20 cases of CIN specimens. HPV typing by restriction enzyme analysis revealed that HPV-16 sequence was present in 11 cases, HPV-18 in 1 case, HPV-31 in 4 cases, HPV-33 in 1 case, HPV-52 in 2 cases, HPV-58 in 3 cases, and an unidentified type(s) in 3 cases. There were 4 cases of mixed infections. This procedure obviates the use of hybridization- based for-mat for identification of at least 8 types of HPV sequences present in a small fraction of cells within a heterogeneous population

ショチュウトウ キョウイク ニホンゴ キョウシ ケンシュウ ニ オケル キョウジュホウ ジュギョウ ニ ツイテ

初・中等教育段階の日本語教育は、各国・地域の外国語教育政策に基づいて展開されており、シラバス、カリキュラム、教材もそれぞれの国・地域で統一されている場合が多く、学習者の発達段階など多くの要因を考慮しながら授業を組み立てていく必要がある。初・中等教育の教師を対象とした研修の教授法授業では、このような背景を考慮した実践的な教授活動を取り入れていくことが重要である。2003年度海外日本語教師短期研修(春期)(以下、春短期研修)における教授法授業では以下の試みを行った。できる限り教授環境を共有する国・地域別のクラス編成を重視し、多国籍のクラスの人数を少数になるようにした。さらに、それぞれの教授環境を重視し、その一方で国・地域を超えた初・中等教育に共通する課題をその枠組みの中で取り上げることとした。 共通に扱う項目としては、それぞれの教師が普段行っているであろう教授活動の流れ、「教授目標設定→授業設計→授業準備・教材作成→授業実施→授業に対する評価」の流れを教授法授業で経験させること、コンピュータの利用、高校訪問という要素を、それぞれの教師の望む程度に応じて教授活動の中に織り込ませることであった。 本稿では、5クラスそれぞれの教授法授業の実際を報告し、コンピュータ利用、高校訪問については全体について報告した。研修に参加したほとんどの教師は、教授法授業のクラス分け、扱った内容について役立ったとしているが、違う国・地域の教師との情報交流の機会をより増やすことを望む声が多かった。さらに、コンピュータ利用を勧めたことについては、IT普及の如何に関わらず好評であった。高校訪問については、半数程度が役立つとしているが、問題があったとする意見もあり、教授法授業への取り入れ方については検討を要する。今後も、初・中等教育に携わる日本語教師への研修を改善していくためには、実践に根ざした情報の蓄積が必要である

A novel mitochondrial DnaJ/Hsp40 family protein BIL2 promotes plant growth and resistance against environmental stress in brassinosteroid signaling

Funding Information: Acknowledgments We thank Dr. Tsuyoshi Nakagawa (Shimane University) for the gift of the gateway vectors, pGWB2, pGWB80, pGWB5, and pGWB3. This work was supported in part by funding from the Program for Promotion of Basic Research Activities for Innovation Bioscience (PROBRAIN) to T.N. and T.A., and CREST, Japan Science and Technology Agency to T.N. and T.A.Plant steroid hormones, brassinosteroids, are essential for growth, development and responses to environmental stresses in plants. Although BR signaling proteins are localized in many organelles, i.e., the plasma membrane, nuclei, endoplasmic reticulum and vacuole, the details regarding the BR signaling pathway from perception at the cellular membrane receptor BRASSINOSTEROID INSENSITIVE 1 (BRI1) to nuclear events include several steps. Brz (Brz220) is a specific inhibitor of BR biosynthesis. In this study, we used Brz-mediated chemical genetics to identify Brz-insensitive-long hypocotyls 2-1D (bil2-1D). The BIL2 gene encodes a mitochondrial-localized DnaJ/Heat shock protein 40 (DnaJ/Hsp40) family, which is involved in protein folding. BIL2-overexpression plants (BIL2-OX) showed cell elongation under Brz treatment, increasing the growth of plant inflorescence and roots, the regulation of BR-responsive gene expression and suppression against the dwarfed BRI1-deficient mutant. BIL2-OX also showed resistance against the mitochondrial ATPase inhibitor oligomycin and higher levels of exogenous ATP compared with wild-type plants. BIL2 participates in resistance against salinity stress and strong light stress. Our results indicate that BIL2 induces cell elongation during BR signaling through the promotion of ATP synthesis in mitochondria.Peer reviewe

Role of Abscisic Acid in Flood-Induced Secondary Aerenchyma Formation in Soybean (Glycine max) Hypocotyls

Phellogen (cork cambium) usually produces cork tissue, but when flooded it produces secondary aerenchyma, comprising living cells with non-suberized walls in the stems, roots, and root nodules of some Fabaceae. In the cell walls of cork tissues, the plant hormone abscisic acid (ABA), promotes suberin deposition. Thus, ABA may decrease in flooded tissues, where secondary aerenchyma cells are developing. Here, we investigated whether ABA is involved in the formation of aerenchyma in soybean (Glycine max) hypocotyls when flooded. Hypocotyls flooded with water produced a large amount of secondary aerenchyma, and were highly porous. On the other hand, application of 1.0 μM ABA suppressed the enlargement of phellogen-derived cells, thereby suppressing subsequent gas space formation, and then almost completely inhibited aerenchyma development. Berberine-aniline blue staining indicated that not only elongated cells in the secondary aerenchyma but also packed cells, which were formed under flooding with ABA, contained no suberized cell walls. Compared to non-flooded plants, the endogenous ABA concentration in the flooded hypocotyls was decreased to 50% within 24 hr, and the low level was maintained for at least 72 hr. In addition, phellogen developed at 48 hr after flooding and secondary aerenchyma was observed at 72 hr. These results indicate that secondary aerenchyma formation requires a decrease in negative regulator ABA in soybean plants, that is, ABA inhibits elongation of cells derived from phellogen in secondary aerenchyma formation such as internodal cell elongation of floating rice stems

Nitrogen Utilization in the Supernodulating Soybean Variety “Sakukei 4” and Its Parental Varieties,“Enrei” and “Tamahomare”

The supernodulating variety “Sakukei 4”, which has improved growth and yield, was recently developed. To evaluate its physiological traits related to the high productivity, we compared dry matter production, nitrogen (N) accumulation and N utilization in Sakukei 4 with those in the parental normally nodulating varieties, Enrei and Tamahomare, and a nonnodulating line, En1282, in pot experiments. The seed yield of Sakukei 4 was similar to that of Enrei and Tamahomare.Leaf area and relative ureide abundance in xylem sap were maintained for longer growth period in Sakukei 4 and Tamahomare than in Enrei. The total amount of nodule N2fixation at maturity was also larger in Sakukei 4 andTamahomare than in Enrei and En1282. In En1282 and Enrei, a larger part of N accumulated in leaves and stems before seed maturation was translocated to seeds during seed filling. However, Sakukei 4 and Tamahomare, more N tended to remain in leaves and stems at maturity. These observations suggested that the physiological traits of dry-matter accumulation and N utilization in Sakukei 4 were quite similar to those in Tamahomare, despite the difference in nodulation. N use efficiency for dry matter production (NUED) was lower in Sakukei 4 than in Enrei and Tamahomare, probably due to the energy cost for sustaining a greater number and mass of nodules. This observation indicated that the low NUED of Sakukei 4 impairs its productivity and seed yield, and therefore Sakukei 4 could not exceed Enrei or Tamahomare in seed productivity