Lysosomal Enzyme Release from Guinea Pig Polymorphonuclear Leukocytes by Influenza Virus

Extracellular release and subsequent decrease of intracellular lysosomal enzymes of guinea pig polymorphonuclear leukocytes (PMNLs) by influenza virus were observed. Total enzyme activity was also decreased in all the enzymes assayed. After incubation of the PMNLs with influenza virus at 37°C for 20 min, the degree in decrease of total enzyme activity varied from enzyme to enzyme: 50% in myeloperoxidase (MPO), 20% in acid phosphatase, 10% in N-Ac-β-glucosaminidase and β-glucuronidase and 5% in lysozyme, respectively. Since MPO assumed to play a critical role in chemiluminescent response of luminol-enhanced system, the dysfunction in oxidative metabolism of PMNLs induced by influenza virus seems to be attributed to intraphagosomal and/or extracellular inactivation of MPO of PMNLs during the process of direct stimulation of respiratory burst

Adherence of Protease-Deficient Mutants of Pseudomonas aeruginosa to a Rabbit Cornea Cell Line (SIRC) Cells

Seven protease-deficient mutants were isolated from Pseudomonas aeruginosa strain IFO 3455 which was mutagenized with nitrosoguanidine. Characterization of these mutants in vitro revealed that all mutants showed pleiotropic changes in the production of other extracellular substances. Among the mutants, two were chosen for a bacterial adherence test to Rabbit Cornea Cell Line (SIRC) cells. One mutant (IFO 3455-2) completely lost its protease activity. Another (IFO 3455-3) retained a low protease activity and was relatively similar to the parental strain with respect to extracellular products except for protease. Both mutants gave not a marked but a slight decrease of adherence as compared with the parental strain. This finding suggests that besides protease more factors are involved in the adhesion between P. aeruginosa and SIRC cells

Physical and Chemical Factors Affecting the Adherence of Pseudomonas aeruginosa to a Rabbit Cornea Cell Line (SIRC) Cells

Adherence of Pseudomonas aeruginosa to SIRC cells was examined by the use of 14C-lysine labeled organisms. Pretreatment of P. aeruginosa with heating, 3% formaldehyde, or ultraviolet caused a significant decrease in adherence to SIRC cells, whereas that with lipase, hyaluronidase, trypsin or protease did not. Treatment of SIRC cells with trypsin, protease, lipase or neuraminidase did not influence the adherence of P. aeruginosa to the cell. Treatment of P. aeruginosa with mannose or galactose inhibited the adherence, while that with fructose, lactose or glucose did not. Treatment of SIRC cells with galactosidase or mannosidase reduced the adherence of the organism. No correlation was demonstrated between the adhering ability and hydrophobicity of P. aeruginosa. The results suggest that both the viability in bacterial site and mannose and/or galactose molecules in cellular site are closely connected with the adherence of P. aeruginosa to SIRC cells

Use of the Fluorescent Staining Method for Determining the Viability of Mycobacterium lepraemurium

The cell suspension of Mycobacterium lepraemurium was exposed to heating of 40°C to 70°C for various lengths of time. The percent green-fluorescent cell by the midified fluorescein diacetate (FDA) -ethidium bromide (EB) staining method was calculated and compared with the infectivity to mice. The reduction in the percentage was associated significantly with the loss of the infectivity

Light and Electron Microscopic Studies on the Attachment of Ureaplasma urealyticum to Human Leukemia Cell

Interaction between human leukemia cell and Ureaplasma urealyticum was investigated light and electron microscopically. The cells adhered with their pseudophodia to the surface of colonies of Ureaplasma urealyticum. A large number of pleomorphic Ureaplasma urealyticum adhered to the cell surface and were seen in the intracellular vacuoles. Ureaplasma urealyticum was captured by the villus-like structure of the cells and actively phagocytized

Development of unit for elective subject from fifth to ninth grade to improve cooperative creation (3)

本研究は, 「21世紀型の教科学力」の新たな観点としての「協同的創造力」の育成をめざして, 自分たちで新たな文化を創造する子どもを育てる協同的創造学習のあり方について実証的に研究を進め, 単元モデルと評価方法を開発することを目的としている。そこで, 教科学習を「協同的創造学習」としてとらえ直すとともに, 中学校での従来の選択教科の時間に加えて, 小学校第5・6学年合同の選択教科の時間を新設して「協同的創造力」を特化して育むことにし, 本年度は, 選択教科の単元モデルの充実・改善と評価方法の確立に取り組んだ。その結果, 選択教科において, これまで開発した単元モデルをより充実させたり, 新たな単元モデルを開発したりすることができた。また, 評価の観点を整理し, 子どもの意識調査やカリキュラム評価に継続して取り組むことによって, 子どもの思いを汲み取り単元を見直していくことができた。今後も必修教科と選択教科のつながりや関連性, 各学年の系統性を整理するとともに, 協同的創造力育成の手だてを整理し, 来年度に向けて, これまで培ったものを生かす新たな学習開発を模索していきたいと考えている

4-Methylumbelliferyl-oleate を用いるモルモット腹腔マクロファージのリバーゼ活性測定法

Some basic conditions for fluorometric measurement of lipase activity of guinea pig peritoneal macrophages were investigated using 4-methylumbelliferyl-oleate as a substrate. The most adequate condition for the assay is as follows: i ) Enzyme preparation recommended is the supernatant of ultrasonicate after freezing and thawing of guinea pig peritoneal macrophages suspended in distilled water. ii) Final concentration of buffered substrate should be adjusted at 0.1 mM in acetate buffer, pH 4.5, free from Triton X-100. iii) Reaction is terminated by addition of 50 mM Tris-HCl buffer, pH 8.6, instead of 50 mM glycine buffer, pH 10.4

ノイラミニダーゼ活性の螢光比色法を用いるノイラミニダーゼ阻止試験の試み

The fluorometric neuraminidase assay method using 4-methylumbelliferyl-N-Ac-α-D-neuraminide as a substrate was investigated for the purpose of ascertaining its validity as a procedure for influenza viral neuraminidase antibody inhibition test. The enzyme antibody inhibition titer obtained by the fluorometric assay system was far weaker than that obtained by the standard colorimetric neuraminidase assay method using fetuin as a substrate. Although solubilization of viral neuraminidase spikes with Triton X-100 increased the enzyme inhibition of antiserum to some extent, there is little possibility of using the fluorometric assay system for the purpose

Depression of Luminol-Enhanced Chemiluminescence of Guinea Pig Polymorphonuclear Leukocytes by Influenza Virus with Special Reference to Neuraminidase

Effect of influenza virus on luminol-enhanced chemiluminescent response of guinea pig polymorphonuclear leukocytes (PMNL) was investigated in order to elucidate a possible mechanism by which influenza virus modified the oxidative metabolism of PMNL. Influenza viruses, strains A/USSR/92/77(H1N1) and B/Kanagawa/3/76(B), rapidly stimulated oxidative burst of resting PMNL and the peak chemiluminescence initiated by them was 5 to 15 times as high as that induced by PBS(—), while the peak chemiluminescent response induced by phorbol myristate acetate (PMA) delayed and the values were counted to be approximately 40% of that of PBS(—)-incubated cells. No significant changes were observed in both activities to enhance chemiluminescent response of resting PMNL and to depress PMA-induced chemiluminescent response when influenza virus with heatinactivated neuraminidase activity at 56°C for 20 min was used. Moreover, neither initiation of oxidative burst of PMNL nor depression of PMA-induced chemiluminescent response was observed with purified neuraminidase extracted from influenza virus. It seems likely that viral neuraminidase is at least not a critical component to modify the oxidative metabolism of guinea pig PMNL

鼠癩菌の培養細胞への付着

Adherence of Mycobacterium lepraemurium to tissue culture cells was examined and compared with that of M. microti. M. lepraemurium used in the present study was maintained on 1% Ogawa egg yolk medium. This microbe adhered to HEp-2 cells much more than A31 or McCoy cells, though the adherence was unusually low as compared with the large infectious dose. Frequency distribution of the number of bacteria agreed nearly with Polya-Eggenberger distribution. The pretreatment of M lepraemurium with heat or protease increased the adherence of the bacteria to HEp-2 cells, whereas the pretreatment with lipase or hyaluronidase retained the adhering ability. The pretreatment of M. microti with heat or protease produced the same effect on the adherence as that of M. lepraemurium. These results suggest that adherence of M. lepraemurium to HEp-2 cells is prevented by protein-like material on the surface of the bacteria, and that the adherence is independent of specific adhesin-receptor interaction