Measurement of intracellular pH by flow cytometry using pH sensitive fluorescence dye, and influence of hyperthermia and amiloride derivatives on the intracellular pH

エールリッヒ腹水癌細胞とそのアドリアマイシン耐性細胞において蛍光pH指示薬2'、7'-bis-(2-carboxyethyl) carboxyfluorescein] (BCECF) の蛍光量をフローサイトメトリーで測定することによって細胞内pHの検量曲線を作成することができた。このことより、これらの細胞においてBCECFの蛍光量で細胞内pHの変化を簡易に比較できることを示唆した。さらに、温熱、Na(+)/H(+) exchanger の阻害例であるアミロライド[3,5-diamino-6-chloro-N-(diaminomethylene) pyrazinecarboxamide]、およびアミロライド誘導隊MH-12-43[N-amidino-3-amino-6-chloro-5-(N-ethyliso-propylamino) pyrazinecarboxyamide] の細胞内pHへの影響をエールリッヒ腹水癌細胞で観察した。37℃では、0.5mMアミロライド、0.05mM　MH-12-43により細胞内pHは減少し、42℃処理によりさらに減少した。42℃において、0.05mM　MH-12-43による細胞内pHの減少は、0.5mMアミロライドによる減少より大きかった。We examined relationship between intensity of intracellular fluorescence of [2', 7'-bis-(2'-carboxyethyl) carboxyfluorescein] (BCECF) and intracellular pH in Ehrlich ascites tumor cells and their adriamycin-resistant strain, and found a good correlation between them at both strains. This suggests that changes in the intracellular pH on these strains may be obtained through measurement of intracellular fluorescence of BCECF by flow cytometry. Further, we examined influence of hyperthermia, 3, 5-diamino-6-chloro-N-(diaminomethylene)pyrazinecarboxamide (amiloride), an inhibitor of Na(+)/H(+) exchanger, and its derivative; N-amidino-3-amino-6-chloro-5-(N-ethylisopropylamino) pyrazinecarboxyamide (MH-12-43) on the intracellular pH in Ehrlich ascites tumor cells. The treatment of 0.5mM amiloride or 0.05mM MH-12-43 reduced intracellular pH at 37℃, while the more reduction was observed by the treatment at 42℃. The reduction of intracellular pH by 0.05mM MH-12-43 was more substantial than that of 0.5mM amiloride at 42℃

Meteor Radar Study of Ionospheric Wind at Kyoto

This paper describes the results of meteor radar observations which have been carried out at Kyoto University since December, 1977. The radar, a coherent pulse doppler radar with a transmitting frequency of 31.57 MHz and a nominal peak power of 10 kW, is able to detect wind fields at meteor regions of 80-110 km. The solar semidiurnal tide generally dominates the wind oscillations at these heights, while a diurnal tide of comparable magnitude reveals itself in a less regular manner. Some anomalies in the amplitude and phase characteristics of the tides are shown, which might be related to a possible interaction with winds and waves, or to some disturbed conditions in the earth's atmosphere. Also planetary scale waves such as quasi-2-day oscillations and resonantly magnified gravity-mode waves have also been found there. Hence, the meteor radar, together with its cooperation with other observations, will helps us understand the dynamical process of atmospheric waves on a global scale

Cepharanthin Reduces Thermotolerance by Enhancing Thermosensitivity in NIH3T3 Cells

The effects of cepharanthin (Ce), glycyrrhizin (G), verapamil (V), and G plus V on induced thermotolerance in NIH3T3 cells were studied. Cells were heated with or without the drug at 45 degrees C for 20 min (the first heating), incubated at 37 degrees C for 12h (the incubation period), and heated again at 45 degrees C for 0-210 min (the second heating). G and V were added throughout the experiment, while Ce was added throughout the experiment or during only the first or second heating, or the incubation period. The cells were harvested after the second heating to evaluate cell survival. In control experiments without any drug, thermotolerance developed and reached the highest peak in the cells incubated for 12h at 37 degrees C. However, thermotolerance in the control cells was suppressed by incubating them at 0 degree C, but developed by subsequent incubation at 37 degrees C. This suggests that the acquisition of thermotolerance by the cells required metabolic processes during the incubation at 37 degrees C. When each drug was present throughout the experiment, only Ce or the combined use of G and V was effective in reducing thermotolerance. Thermotolerance was also suppressed in the presence of Ce during the second heating. These results indicate that Ce reduces thermotolerance by enhancing thermosensitivity rather than by inhibiting the development of thermotolerance.</p