High Time-Resolution Monitoring of Free-Tropospheric Sulfur Dioxide and Nitric Acid at the Summit of Mt. Fuji, Japan

This is the first paper that describes the atmospheric sulfur dioxide (SO2) and nitric acid (HNO3) monitored with a good time-resolution at the summit (3,776 m a.s.l.), which is located in the free troposphere, and southeastern foot (1,284 m a.s.l.) of Mt. Fuji. Japan. During the summer of 2012, two analytical systems consisting mainly of a parallel-plate wet denuder and ion chromatograph operated simultaneously at both the sampling sites. All the samples collected at both the sampling sites contained detectable levels of sulfate from gas-phase SO2 while the nitrate from gas-phase HNO3 was detectable in 97.8% of air samples at the southeastern foot and 88.4% at the summit. The average concentrations of SO2 and HNO3 were, respectively, 0.061 ± 0.071 and 0.031 ± 0.020 ppbv at the summit (n = 672), and 0.347 ± 0.425 and 0.146 ± 0.070 ppbv at the southeastern foot (n = 1344) of Mt. Fuji. Both the acidic gases at the southeastern foot and the HNO3 at the summit showed a diurnal pattern with daytime maxima and nighttime minima. Meanwhile, the SO2 at the summit did not show a distinct shift, which indicates the SO2 concentrations at the summit would be principally controlled by the advection of air parcel in the free troposphere

Downregulation of CD4 is required for maintenance of viral infectivity of HIV-1

AbstractDownregulation of virus receptors on the cell surface is considered to be important in preventing superinfection. HIV-1 encodes multiple gene products, Env, Vpu, and Nef, involved in downregulation of CD4, a major HIV-1 receptor. We found that simultaneous mutations in both vpu and nef severely impaired virus replication. We examined the involvement of CD4 downregulation mediated by Vpu and Nef in the modification of virus infectivity. The mutation in vpu increased CD4 incorporation into virions without affecting the Env content in it, inhibiting the attachment step of virions to the CD4-positive cell surface. Although a single mutation in nef suppresses virus infectivity via a CD4-independent mechanism, it could augment CD4 incorporation in virions in combination with a vpu mutation. These results indicated that CD4 downregulation was necessary for maintenance of Env function in the virion

Characteristics of a Monoacylglycerol Lipase Isolated from Pseudomonas sp. LP7315 -Hydrolysis and Synthesis of Monoglycerides

A monoacylglycerol lipase (MGL) was purified from Pseudomonas sp. LP7315 by ammonium sulfate precipitation, anion-exchange chromatography, and preparative electrophoresis. The purified enzyme was homogeneous on an SDS-polyacrylamide gel with a molecular mass of 59 kDa. Its hydrolytic activity was confirmed to be specific for monoglycerides: the enzyme did not hydrolyze diand triglycerides. MGL was found to be stable even after l-h incubation at 65℃. The hydrolytic activity depended not only on temperature and pH but also on the type of monoglyceride used. MGL also catalyzed monoglyceride synthesis at 65℃ in a solvent-free two-phase system, in which fatty acid droplets were dispersed in the glycerol phase with a low water content. The synthetic reaction proceeded at a constant rate for approximately 24 h and reached an equilibrium after 48 h of reaction. The initial rate of the synthetic reaction depended on several factors: the type of fatty acid used as the substrate, the amounts of fatty acid and glycerol, and the concentration of MGL in the glycerol phase. To analyze the effects of these factors, a kinetic model was developed based on the assumption that the adsorption equilibrium of MGL molecules at the interface between the two phases is the rate-determining factor for the synthetic reaction. The model was found to yield a good approximation of the initial synthetic rate under various reaction conditions. The analysis suggests that the adsorption behavior of MGL onto the interface had a large effect on the initial rate of the monoglyceride synthesis

Mutations in N-terminal flanking region of blue light-sensing light-oxygen and voltage 2 (LOV2) domain disrupt its repressive activity on kinase domain in the Chlamydomonas phototropin.

Phototropin is a light-regulated kinase that mediates a variety of photoresponses such as phototropism, chloroplast positioning, and stomata opening in plants to increase the photosynthetic efficiency. Blue light stimulus first induces local conformational changes in the chromophore-bearing light-oxygen and voltage 2 (LOV2) domain of phototropin, which in turn activates the serine/threonine (Ser/Thr) kinase domain in the C terminus. To examine the kinase activity of full-length phototropin conventionally, we employed the budding yeast Saccharomyces cerevisiae. In this organism, Ser/Thr kinases (Fpk1p and Fpk2p) that show high sequence similarity to the kinase domain of phototropins exist. First, we demonstrated that the phototropin from Chlamydomonas reinhardtii (CrPHOT) could complement loss of Fpk1p and Fpk2p to allow cell growth in yeast. Furthermore, this reaction was blue light-dependent, indicating that CrPHOT was indeed light-activated in yeast cells. We applied this system to a large scale screening for amino acid substitutions in CrPHOT that elevated the kinase activity in darkness. Consequently, we identified a cluster of mutations located in the N-terminal flanking region of LOV2 (R199C, L202L, D203N/G/V, L204P, T207I, and R210H). An in vitro phosphorylation assay confirmed that these mutations substantially reduced the repressive activity of LOV2 on the kinase domain in darkness. Furthermore, biochemical analyses of the representative T207I mutant demonstrated that the mutation affected neither spectral nor multimerization properties of CrPHOT. Hence, the N-terminal flanking region of LOV2, as is the case with the C-terminal flanking Jα region, appears to play a crucial role in the regulation of kinase activity in phototropin

Mechanical properties of epithelial cells in domes investigated using atomic force microscopy

As epithelial cells in vitro reach a highly confluent state, the cells often form a microscale dome-like architecture that encloses a fluid-filled lumen. The domes are stabilized by mechanical stress and luminal pressure. However, the mechanical properties of cells that form epithelial domes remain poorly characterized at the single-cell level. In this study, we used atomic force microscopy (AFM) to measure the mechanical properties of cells forming epithelial domes. AFM showed that the apparent Young’s modulus of cells in domes was significantly higher when compared with that in the surrounding monolayer. AFM also showed that the stiffness and tension of cells in domes were positively correlated with the apical cell area, depending on the degree of cell stretching. This correlation disappeared when actin filaments were depolymerized or when the ATPase activity of myosin II was inhibited, which often led to a large fluctuation in dome formation. The results indicated that heterogeneous actomyosin structures organized by stretching single cells played a crucial role in stabilizing dome formation. Our findings provide new insights into the mechanical properties of three-dimensional deformable tissue explored using AFM at the single-cell level

Practical Approaches based-on New Course of Study for Japanese Language Education by MEXT : Japanese language learning for developing students' world of language through collaborative leaning.

本研究は, 学習者が自分のまわりの他者と出会いかかわり合うことをとおして, 自分自身の言葉の世界を広げ, 自らの学びを探究していくことができるようにするための効果的な指導法を開発していくことを目的としている。そこで, 「さまざまな他者と出会い, 問いを立てる」「個の考えを深め吟味する」「他者とともによりよい方法で問題解決に取り組む」「学びの成果を適切に表現する」「学びを振り返り新たな学びにつなげる」という学習のプロセスに留意して単元構成を行った。2年生では, 自分の発想を言葉として表現したものを, 身近な他者に伝える経験を積ませる学習活動を構成することによって, 達成感を感じさせることができた。また, 子どもたちの意欲を喚起し学習に見通しをもたせることで, 発想豊かにお話作りに取り組ませることができた。3年生では, 説明文の内容を地域の行事と比べながら読ませたり, 今まで何気なく経験してきている行事の意味を改めて見つめ直させたりすることができた。また, 既習教材との比較によって, 筆者の意図や論理構成の工夫に気付かせることができた。今後も, 言葉の世界をひらき, 学びを自ら探求する学習のあり方を探っていきたい