Testing the Anomaly Mediation at the LHC

We consider a supersymmetric model in which gaugino masses are generated by the anomaly-mediation mechanism while scalar masses are from tree-level supergravity interaction. In such a model, scalar fermions as well as Higgsinos become as heavy as O(10-100TeV) and hence only the gauginos are superparticles kinematically accessible to the LHC. We study how and how well the properties of gauginos can be studied. We also discuss the strategy to test the anomaly-mediation model at the LHC.Comment: 12 pages, 6 figure

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℃

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

加温後のtsAF8細胞の細胞周期

Thermotolerance in tsAF8 cells develops during incubation at 34℃ after heating at 45℃, while it is suppressed by the following incubation at a non-permissive temperature of 39.7℃ after the same heating. The incubation temperature after heating may affect the cell cycle and consequently thermotolerance. In the present study, a relationship between the thermotolerance and the cell cycle of tsAF8 was investigated. The cell cycle fractions and DNA synthesis were measured by flow cytometry using double staining with propidium iodide and bromodeoxyuridine. When the tsAF8 cells were heated at 45℃ for 20 min, and thereafter incubated at 34℃, bromodeoxyuridine uptake in the S phase cells (DNA synthesis) was recovered to 65.1% 6 h after the heating, and the cells showed gradual accumulation in the G(2)/M phase. When the cells were incubated at 39.7℃ after heating at 45℃ for 20 min, then showed inhibition of thermotolerance development, the DNA synthesis was recovered to 15.1% temporarily 6 h after the heating, but it became 0% after 12 h, and the cells did not remarkably accumulate in any phases of the cell cycle. This inhibition of DNA synthesis at 39.7℃ was considered to be the result of cell survival decreasing by a step-down heating. However, the relationship between the thermotolerance and the cell cycle was not found out in tsAF8 cells, because the cells did not accumulate in any phases of the cell cycle under the inhibitory condition of thermotolerance.tsAF8細胞は45℃の加温後34℃で培養すると温熱耐性が速やかに発現するが,加温後,制限温度である39.7℃で培養すると温熱耐性の発現が抑制される｡加温後の培養温度が細胞周期に影響し,その結果として温熱耐性発現に影響を与えている可能性があることから,今回,Propidium Iodide(PI)とbromodeoxyuridine(BrdU)でtsAF8細胞を二重染色し,フローサイトメトリーによって温熱耐性と細胞周期の関係の有無について調べた｡tsAF8細胞を45℃20分の加温後34℃で培養すると,6時間後にはG(1)期の細胞が減少し,12時間後にはG(2)/M期への蓄積が見られた｡しかし,加温後39.7℃で培養した場合には細胞周期の進行がほとんど見られなかった｡BrdU の取込みは,加温せずに39.7℃で培養した場合には活発に行われ,また,45℃20分加温後34℃で培養した場合には,6時間後にはBrdUの取り込みは65.1%まで回復した｡しかし,温熱耐性発現の抑制が観察される45℃20分加温後39.7℃で培養した場合には,BrdUの取込み量は6時間後に一時的に15.1%に回復するが,12時間後には取込み量はゼロとなった｡BrdUの取り込みが阻害されたのはstep-down heatingの現象による細胞生存率の減少が原因だと考えられたが,温熱耐性発現の抑制が観察される条件下では細胞周期の特定の時期への集積がなかったことから,温熱耐性と細胞周期との関係はtsAF8細胞においては見い出されなかった

Relationship between intracellular uptake of adriamycin and membrane potential in ADR resistant Ehrlich ascites tumor cells

We observed adiamycin (ADR) uptake and cellular transmembrane potential [amount of intracellular fluorescence of 3,3'- (Di-n-hexyl)- 2,2'- oxacarbocyanine iodide (NK-2280)] in ADR-resistant cells established from Ehrlich ascites tumor cells (EATC) and wild type EATC. In ADR-resistant cells, ADR uptake and the cellular transmembrane potential decreased as the degree of resistance increased. 4,4'- diisothiocyanatostilbene- 2,2'- disulfonic acid (DIDS) induced markedly decreases of ADR uptake and the cellular transmembrane potential. A good correlation was observed between ADR uptake and transmembrane potential in cultured cells