• Archives of Pharmacal Research
• /
• v.21 no.1
• /
• pp.46-53
• /
• 1998

When NIH3T3 cells were exposed to mild heat and recovered at $37^{\circ}C$ for various time intervals, they were thermotolerant and resistant to subsequent stresses including heat, oxidative stresses, and antitumor drug methotrexate which are apoptotic inducers. The induction kinetics of apoptosis by stresses were determined by DNA fragmentation and protein synthesis using $[35^S]$methionine pulse labeling. We investigated the hypothesis that thermotolerant cells were resistant to apoptotic cell death compared to control cells when both cells were exposed to various stresses inducing apoptosis. The cellular changes in thermotolerant cells were examined to determine which components are involved in this resistance. At first, the degree of resistance correlates with the extent of heat shock protein synthesis which were varied depending on the heating times at $45^{\circ}C$ and recovery times at $37^{\circ}C$after heat shock. Secondly, membrane permeability change was observed in thermotolerant cells. When cells prelabeled with $[^{3}H]$thymidine were exposed to various amounts of heat and recovered at $37^{\circ}C$ for 1/2 to 24 h, the permeability of cytosolic $[^{3}H]$thymidine in thermotolerant cells was 4 fold higher than that in control cells. Thirdly, the protein synthesis rates in thermotolerant and control cells were measured after exposing the cells to the same extent of stress. It turned out that thermotolerant cells were less damaged to same amount of stress than control cells, although the recovery rates are very similar to each other. These results demonstrate that an increase of heat shock proteins and membrane changes in thermotolerant cells may protect the cells from the stresses and increase the resistance to apoptotic cell death, even though the exact mechanism should be further studied.

• Journal of The Korean Society of Grassland and Forage Science
• /
• v.17 no.3
• /
• pp.249-256
• /
• 1997

A pulse-chase labeling of $^{15}N$ on winter rye (Scale cereale) and forage rape (Brassica napus) grown at $15^{\circ}C$ and $25^{\circ}C$ was carried out to determine the effects of low temperature on the uptake exogenous N and the remobilization of endogenous N. The growth rate of leaves and roots depressed at $5^{\circ}C$. AAer 9 days at $5^{\circ}C$, nitrogen content of leaves decreased to 20% on the average while that of roots increased to 12% compared with the plants grown at $25^{\circ}C$. Total content of $NO_3$- uptake 60m medium was 23.0 and 43.5 mg Nlplant, respectively, for winter rye and forage rape grown at $5^{\circ}C$ during 9 days. These values were corresponded to 59.3 and 26.1% lower uptake than those of $25^{\circ}C$. A large part of 1 5 ~ was distributed into leaves throughout time course in both of two species. The content of $^{15}N$ in leaves of winter rye at day 6 increased to 166 and 296 $\mu^{15}$N/plant compared with the initial value (day 0) in the plants grown at $5^{\circ}C$and $25^{\circ}C$ , corresponding to 90 and 163 $\mu$g N of remobilization h m roots into leaves during the fist 6 days. From 7 to 9 days, 75 and 52 $\mu$gN of outflow 6om leaves were occurred at $5^{\circ}C$ and $25^{\circ}C$. However, little remobilization of endogenous N was estimated in forage rape throughout the entire time course regardless of temperature treatment. Comparing two species studied, winter rye was much sensitively influenced by low temperature on the uptake of exogenous N and the remobilization of endogenous N.

• Proceedings of the KSAR Conference
• /
• 2003.06a
• /
• pp.22-22
• /
• 2003

This study was conducted to determine effects of polyvinyl alcohol (PVA), fetal bovine serum (FBS) and bovine serum albumin (BSA) on blastocoel formation, cell number, apoptosis and apoptosis-related gene expression of porcine diploid parthenotes developing in vitro. Embryos were collected from 2-cell or late 4-cell diploid parthenotes that activated with electro pulse, and in vitro cultured in the NCSU 23 medium supplemented without or with 0.1% PVA, 10% FBS or 0.4% BSA for day 7. The morphological analysis of apoptosis in embryos was carried out using propidium iodide staining and terminal deoxynucleotidyl transferase mediated dUTP nick end labeling. The expressions of Bcl-xL, Bak and P53 in blastocyst stage parthenotes and in vivo-derived blastocysts were determined using semiquantitative RT-PCR. The addition of 0.4% BSA to the culture medium enhanced the development of 2- or late 4-cell stage parthenotes to the blastocysts stage (P < 0.01) while FBS decreased the incidence of blastocoel formation. FBS also reduced cell numbers of blastocysts developed from both 2- (P < 0.001) and late 4-cell (P < 0.05) embryos and increased percentage of apoptosis in the blastocysts (P < 0.001). The relative abundance of Bcl-xL mRNA in diploid parthenotes cultured from 2-cell stage in the presence of BSA is similar with that in in vivo derived embryos, but is significantly higher than in parthenotes cultured with FBS, PVA or none protein supplement control. Bak mRNA showed a significant increase at the blastocyst stage in FBS supplement medium. This result suggests that apoptosis related gene expression is significantly affected by protein supplements, which may result in alteration of apoptosis and embryo viability of porcine embryos developing in vitro.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.31 no.4
• /
• pp.672-678
• /
• 2002

Oxidative stress contributes to cellular injury following clinical and experimental ischemia/reperfusion scenarios. Oxidative injury can induce cellular and nuclear damages that result in apoptotic cell death. We tested the hypothesis that the catechin flavonoid of (-)epigallocatechin gallate, a green tea polyphenol, inhibits hydrogen peroxide ($H_2O$$_2$)-induced apoptosis in human umbilical vein endothelial cells. The effect of apigenin, a flavone found in citrus fruits, on apoptosis parameters was also examined. A 30 min pulse treatment with 0.25 mM $H_2O$$_2$ decreased endothelial cell viability within 24 hrs by > 30% ; this was associated with nuclear condensation and biochemical DNA damage consistent with programmed cell death. In the 0.25 mM $H_2O$$_2$apoptosis model, 50${\mu}{\textrm}{m}$ (-)epigallocatechin gallate markedly increased cell viability with a reduction in the nuclear condensation and DNA fragmentation. In contrast, equimicromolar apigenin increased cell loss with intense DNA laddering, positive nick-end labeling and Hoechst 33258 staining. Thus, polyphenolic (-)epigallocatechin gallate, but not apigenin flavone, qualify as an antioxidant in apoptosis models caused by oxidative stress. Further work is necessary for elucidating the anti-apoptotic mechanisms of polyphenolic catechins.