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Photodynamic stimulation causes sustained increase in intracellular calcium concentration in cells of small cell lung carcinoma
1998
Hayashi, M. (Hokkaido Univ., Sapporo (Japan). Faculty of Veterinary Medicine) | Kanno, T.
Photodynamic agents, due to their selective uptake by tumor cells and photon-dependent selective activation, have immense implications for cancer treatment. The present study provided direct evidence that the photon activation of chloro-aluminum phthalocyanine sulphonate (AlPcS4) in the presence of extracellular Ca2+ caused a rapid increase followed by a sustained increase in intracellular concentration of calcium ion ([Ca2+]i) in a small cell lung carcinoma (SCLC) cell line, SBC-3. The [Ca2+]i increase by photodynamic stimulation was completely inhibited by the removal of extracellular Ca2+ and reintroduction of extracellular Ca2+ immediately led to a rapid elevation of [Ca2+]i. However, the increase was not inhibited by application of Ni2+, nifedipine, or SKandF 96365, a receptor-mediated and voltage-dependent Ca2+ entry blocker. The photosensitizer AlPcS4 alone or light alone (4 min) had no effect on [Ca2+]i. Cytotoxicity examination by trypan blue exclusion test, however, suggested photodynamic stimulation-induced cell injury which was observed in both the presence and the absence of extracellular Ca2+. These results indicate that [Ca2+]i increase may not be mandatory for photodynamic stimulation-induced cell injury. Whether [Ca2+]i increase can accelerate, at least in part, cell death under the physiological condition, whether the mechanism(s) of cell death can be different in the presence and the absence of extracellular Ca2+, and whether [Ca2+]i increase can be totally unrelated to cell death await further work
Показать больше [+] Меньше [-]A monoclonal antibody, 169.1, against canine leukocyte surface antigen identifies cytoskeletal components in epithelial cells and peripheral neurons
1997
Iwami, Y. (Hokkaido Univ., Sapporo (Japan)) | Hashimoto, Y. | Iwanaga, T.
Electrofusion of zona-free mouse embryonic cells in electrolytes and their development in vitro
1995
Elsheikh, A.S. (Hokkaido Univ., Sapporo (Japan)) | Takahashi, Y. | Tanaka, H. | Hishinuma, M. | Kanagawa, H.
The influence of increasing the physical electrofusion parameters, direct current (DC) pulse strength, pulse duration, pulse number, alternating current (AC) voltage and alignment time, in electrolytes on the rates of fusion,degeneration and development of zona-free mouse 2-cell embryos were examined. Furthermore, the effects of physiological saline and mannitol as fusion media and various mouse strains were also evaluated. Dulbecco's phosphate-buffered saline (PBS) supplemented with 10% fetal calf serum was used as the main fusion solution. A significant increase in the rate of fusion (P0.05) was obtained by increasing pulse strength from 30 to 300 V/mm. The embryos fused at the pules strengths of 30 to 70 V/mm had significantly higher development rates to blastocysts compared with those fused at 100 to 300 V/mm (P0.05). There were no significant differences in the rates of fusion, degeneration and d development to blastocysts when the pulse duration was increased from 30 to 90 mu-sec. Although fusion rates were increased (P0.05) by increasing the pulse number up to 4, a significant decrease (P0.05) in development to blastocysts was observed when the pulse number was 5. Application of AC voltage prior to the DC pulse tended to increase the fusion rate (89.2-93.8%), compared with fusion with the DC pulse only (75.0%). Prolongation of alignment time from 5 to 15 sec had no effect on the fusion rate. Under the optimum conditions (2 pulses of DC of 7- V/mm, 70 mu-sec pulse duration and AC of 5 V/mm for 5 sec), no significant difference was obtained in the fusion and development rates in different mouse strains, nor were fusion and development rates significantly different among PBS, physiological saline and mannitol solutions (P0.05)
Показать больше [+] Меньше [-]Cytotoxic effect of acyclovir on cultured mammalian cells to which herpesvirus thymidine kinase gene was introduced
1989
Tanabe, K. (Hokkaido Univ., Sapporo (Japan). Faculty of Veterinary Medicine) | Hiraoka, W. | Kuwabara, M. | Sato, F. | Narita, T. | Niikura, M.
Lymphocyte transformation of peripheral blood lymphocytes and plaque forming cells of the spleen and the mesenteric lymph node in suckling piglets with and without immunopotentiators
1983
Namioka, S. | Ohsugi, T. | Fujimoto, T. | Maeda, Y. (Hokkaido Univ., Sapporo (Japan). Faculty of Veterinary Medicine)
Leptin inhibits hepatocyte growth factor-induced ductal morphogenesis of bovine mammary epithelial cells
2007
Yamaji, D.(Hokkaido Univ., Sapporo (Japan)) | Kamikawa, A. | Soliman, M.M. | Ito, T. | Ahmed, M.M. | Makondo, K. | Watanabe, A. | Saito, M. | Kimura, K.
We examined the effect of stroma-derived factors, hepatocyte growth factor (HGF) and leptin, on morphological differentiation of bovine mammary epithelial cells (BMEC) in collagen gel three-dimensional culture in vitro. BMEC treated with HGF, but not leptin, formed duct-like organoids. The formation of organoids by HGF was enhanced by treatment with a mixture of insulin, cortisol and prolactin, while BMEC treated with the mixture alone did not produce the organoid. In contrast, the formation of organoids by HGF was dose-dependently inhibited by simultaneous addition of leptin, regardless of the presence or absence of the hormone mixture. These results suggest that stroma-derived factors intricately regulate mammary epithelial morphogenesis.
Показать больше [+] Меньше [-]Application of FT-IR and ESR spectroscopic techniques to the study of CCl4-induced peroxidation in rat liver microsomes
2000
Yoon, S. (Hokkaido Univ., Sapporo (Japan)) | Maruyama, Y. | Kazusaka, A. | Fujita, S.
FT-IR and ESR were used for on the investigation of the CCl4-induced peroxidation of rat liver microsomes in combination with biochemical methods. Lipid peroxidation was assayed by TBA reagent in the presence of CCl4 and NADPH. The CCl3 - radical was detected by ESR spectroscopy with a spin trapping reagent of PBN. The FT-IR spectroscopy revealed that absorption band of -C-H in -C=C-H decreased in intensity at 3012 cm(-1), but the absorption bands of the phosphate head and choline in the phospholipids did not significantly change between 1300 and 900 cm(-1). These findings were interpreted to be due to the removal of H- from -C=C-H by radicals as the first step of lipid peroxidation, and to the absence of dephosphorylation of phospholipids in the microsomal membrane. This is the first IR spectroscopic evidence indicating the nature of damage to a microsomal membrane caused by CCl4 treatment. The spectroscopies used here demonstrated that they are useful tools to observe the damage
Показать больше [+] Меньше [-]Postnatal changes in Rho and Rho-related proteins in the mouse brain
2000
Komagome, R. (Hokkaido Univ., Sapporo (Japan)) | Kimura, K. | Saito, M.
To provide information on the role of Rho, a GTP-binding protein, in postnatal development of the brain cells, the change in the levels of Rho protein and Rho-related proteins was examined in the brain of mice for two weeks after birth, in parallel with the changes in the activity of marker enzymes for neuronal and glial cells. The activities of acetylcholine esterase and choline acetyltransferase of whole brain homogenate, both of which are neuronal marker enzymes, were progressively increased in an age-dependent manner. The activity of 2',3'-cyclic nucleotide 3'-phosphohydrolase, a glial marker enzyme, increased markedly between one and two weeks after birth. In contrast, the levels of RhoA and RhoB in the membrane fraction were decreased during the postnatal period. The amount of Rho GDP dissociation inhibitor, a regulatory protein for Rho, was unchanged, while those of Rho target proteins, Rock-2 and citron, were gradually increased. Since the inactivation of Rho is known to induce neurite extension and neuronal and glial differentiation in vitro, our results suggest that the Rho signalling pathway plays a regulatory role in the postnatal differentiation of neuronal and glial cells in vivo
Показать больше [+] Меньше [-]Effects of substance P on nicotine-induced intracellular Ca2+ dynamics in bovine adrenal chromaffin cells
1999
Suzuki, S. (Hokkaido Univ., Sapporo (Japan)) | Habara, Y. | Kanno, T.
Substance P (SP) is colocalized with ACh in splanchnic nerves that innervate into adrenal medulla and the peptide has been shown to inhibit nicotinic agonists-induced catecholamine secretion. To elucidate the effects of SP on cytosolic Ca(2+) dynamics, the present study was conducted using fura-2-loaded isolated bovine adrenal chromaffin cells. Stimulation of the cells with nicotine (10-100mu-M) produced a rapid rise of cytosolic Ca(2+) concentration ([Ca(2+)]i), the peak level of which increased in a dose-dependent manner, followed by a gradual decay. In the presence of 10mu-M SP, the dose-response relationship of the peak levels shifted downward. Quantitative analyses implied that SP inhibits the nicotine-induced Ca(2+) influx in a noncompetitive manner. Nicotinic acetylcholine receptor is composed of two major functional domains: an agonist-binding site and an ionophore or channel domain. Agonist binding activates ionophore / channel domain and causes mainly Na(+) influx. This Na(+) influx depolarizes the cell and activates voltage-dependent Ca(2+) channels. Based on this fact, the present results indicate that SP dose not block nicotine binding sites but interferes with other sites of nicotinic receptor / channel molecule, most probably a channel domain. It was suggested that SP colocalized with ACh in splanchnic nerves functions as a physiological modulator of catecholamine secretion by non-competitively suppressing ACh-induced cytosolic Ca(2+) dynamics in bovine a
Показать больше [+] Меньше [-]Longevity and fecundity of Trichinella spiralis in mast cell-deficient S1/S1d mice
1987
Itayama, H. (Hokkaido Univ., Sapporo (Japan). Faculty of Veterinary Medicine) | Oku, Y. | Kamiya, M.