The leaf of Pinus (P.) densiflora, a pine tree widely distributed in Asian countries, has been used as a traditional medicine. In the present study, we investigated the anticancer activity of essential oil, extracted by steam distillation, from the leaf of P. densiflora in YD-8 human oral squamous cell carcinoma (OSCC) cells. Treatment of YD-8 cells with P. densiflora leaf essential oil (PLEO) at 60 μg/ml for 8 h strongly inhibited proliferation and survival and induced apoptosis. Notably, treatment with PLEO led to generation of ROS, activation of caspase-9, PARP cleavage, down-regulation of Bcl-2, and phosphorylation of ERK-1/2 and JNK-1/2 in YD-8 cells. Treatment with PLEO, however, did not affect the expression of Bax, XIAP and GRP78. Importantly, pharmacological inhibition studies demonstrated that treatment with vitamin E (an anti-oxidant) or z-VAD-fmk (a pan-caspase inhibitor), but not with PD98059 (an ERK-1/2 inhibitor) or SP600125 (a JNK-1/2 inhibitor), strongly suppressed PLEO-induced apoptosis in YD-8 cells and reduction of their survival. Vitamin E treatment further blocked activation of caspase-9 and Bcl-2 down-regulation induced by PLEO. Thus, these results demonstrate firstly that PLEO has anti-proliferative, anti-survival and pro-apoptotic effects on YD-8 cells and the effects are largely due to the ROS-dependent activation of caspases.
The aims of this study were to estimate daily intake of macrominerals from beverages, liquid teas, and liquid coffees and to evaluate their potential health risks for Korean children and adolescents (1-to 19 years old). Assessment of dietary intake was conducted using the actual level of sodium, calcium, phosphorus, potassium, and magnesium in non-alcoholic beverages and (207 beverages, 19 liquid teas, and 24 liquid coffees) the food consumption amount drawn from "The Fourth Korea National Health and Nutrition Examination Survey (2007)(2008)(2009) ". To estimate the dietary intake of nonalcoholic beverages, 6,082 children and adolescents (Scenario I) were compared with 1,704 non-alcoholic beverage consumption subjects among them (Scenario II). Calculation of the estimated daily intake of macrominerals was based on point estimates and probabilistic estimates. The values of probabilistic macromineral intake, which is a Monte-Carlo approach considering probabilistic density functions of variables, were presented using the probabilistic model. The level of safety for macrominerals was evaluated by comparison with population nutrient intake goal (Goal, 2.0 g/day) for sodium, tolerable upper intake level (UL) for calcium (2,500 mg/day) and phosphorus (3,000-3,500 mg/day) set by the Korean Nutrition Society (Dietary Reference Intakes for Koreans, KDRI). For total children and adolescents (Scenario I), mean daily intake of sodium, calcium, phosphorus, potassium, and magnesium estimated by probabilistic estimates using Monte Carlo simulation was, respectively, 7. 93, 10.92, 6.73, 23.41, and 1.11, and 95th percentile daily intake of those was, respectively, 28.02, 44.86, 27.43, 98.14, and 3.87 mg/day. For consumers-only (Scenario II), mean daily intake of sodium, calcium, phosphorus, potassium, and magnesium estimated by probabilistic estimates using Monte Carlo simulation was, respectively, 19.10, 25.77, 15.83, 56.56, and 2.86 mg/day, and 95th percentile daily intake of those was, respectively, 62.67, 101.95, 62.09, 227.92, and 8.67 mg/day. For Scenarios I·II, sodium, calcium, and phosphorus did not have a mean an 95th percentile intake that met or exceeded the 5% of Goal and UL. (Korean J Nutr 2013; 46(1): 50 ~ 60) KEY WORDS: macrominerals, estimated daily intake, non-alcoholic beverage, probabilistic estimates.
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