• Ecology and Resilient Infrastructure
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• v.9 no.4
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• pp.259-268
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• 2022

In this study, seeds of Festuca ovina var. coreana growing in waste coal landfills exposed to heavy metal contamination for a long time were collected, and growth characteristics and heavy metal accumulation capacity were evaluated through greenhouse cultivation experiments with germinated seedlings, and was conducted for the applicability of phytoremediation technology. Concentration gradients of arsenic-treated artificial soil were 25, 62.5, 125, and 250 mg/kg, respectively, lead concentrations were 200, 500, 1000, and 2000 mg/kg, and cadmium concentrations were 15, 30, 60, and 100 mg/kg, respectively In the arsenic, lead, and cadmium-treated experimental groups, the number of leaves of F. ovina var. coreana decreased in all compared to the control group except for the lead-treated groups (200, 500, and 1000 mg/kg). Length growth of the shoot part was increased in all of the arsenic treatment groups compared to the control group, but decreased in all of the root parts. In the 1000 and 2000 mg/kg lead treatment groups, lengths increased compared to the control group, but in the other treatments, they were shorter than the control group. In the case of the cadmium treatment group, all of the shoot parts were increased compared to the control group, and all of the root parts were decreased. In the case of arsenic treatment, the biomass was decreased at all parts and all concentrations compared to the control group. The 200, 500, and 1000 mg/kg lead treatments showed larger biomass than the control group in both shoot and root parts. In the cadmium treatment group, the biomass of both shoot and root parts decreased compared to the control group. As the concentration of heavy metal treatment increased, both the number of leaves and the biomass by plant parts tended to decrease, and the length growth of the shoot part tended to increase slightly, but the root part tended to decrease slightly. The arsenic accumulation concentrations of the shoot and root parts of the 62.5 mg/kg arsenic treatment area were 9.4 mg/kg and 253.3 mg/kg, respectively. While the shoot part of the 250 mg/kg arsenic treatment area withered away, the arsenic accumulation concentration in the root part was analyzed to be 859.1 mg/kg, In the 2,000 mg/kg lead treatment area, the shoot and root parts accumulated 10,308.1 and 11,012.0 mg/kg, which were 1.1 times higher than the root parts. At 100 mg/kg cadmium treatment, the shoot and root parts were 176.0 and 287.2 mg/kg, and the root part accumulated 1.6 times higher than the shoot part. As a result of tolerance evaluation of F. ovina var. coreana, multi-tolerance to three heavy metals was confirmed by maintaining growth without dying in all treatment groups of arsenic, lead, and cadmium. Plant extraction (phytoextraction) of F. ovina var. coreana was verified as a species that can be applied up to 2,000 mg/kg of soil lead contamination.

• Journal of Plant Biotechnology
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• v.5 no.3
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• pp.143-148
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• 2003

Bax, a mammalian pro-apoptotic member of the Bcl-2 family induces cell death when expressed in yeast. To investigate whether Bax expression can induce cell death in plant, we produced transgenic Arabidopsis plants that contained murine Bax cDNA under control of a glucocorticoid-inducible promoter. Transgenic plants treated with dexamethasone, a strong synthetic glucocorticoid, induced Bax accumulation and cell death, suggesting that some elements of cell death mechanism by Bax may be conserved among various organisms. Therefore, we developed novel yeast genetic system, and cloned several Plant Bax Inhibitors (PBIs). Here, we report the function of two PBIs in detail. PBI1 is ascorbate peroxidase (sAPX). Fluorescence method of dihydrorhodamine123 oxidation revealed that expression of Bax in yeast cells generated reactive oxygen species (ROS), and which was greatly reduced by co-expression with sAPX. These results suggest that sAPX inhibits the generation of ROS by Bax, which in turn suppresses Baxinduced cell death in yeast. PBI2 encodes nucleoside diphosphate kinase (NDPK). ROS stress strongly induces the expression of the NDPK2 gene in Arabidopsis thaliana (AtNDPK2). Transgenic plants overexpressing AtNDPK2 have lower levels of ROS than wildtype plants. Mutants lacking AtNDPK2 had higher levels of ROS than wildtype. $H_2O_2$ treatment induced the phosphorylation of two endogenous proteins whose molecular weights suggested they are AtMPK3 and AtMPK6. In the absence of $H_2O_2$ treatment, phosphorylation of these proteins was slightly elevated in plants overexpressing AtNDPK2 but markedly decreased in the AtNDPK2 deletion mutant. Yeast two-hybrid and in vitro protein pull-down assays revealed that AtNDPK2 specifically interacts with AtMPK3 and AtMPK6. Furthermore, AtNDPK2 also enhances the MSP phosphorylation activity of AtMPK3 in vitro. Finally, constitutive overexpression of AtNDPK2 in Arabidopsis plants conferred an enhanced tolerance to multiple environmental stresses that elicit ROS accumulation in situ. Thus, AtNDPK2 appears to playa novel regulatory role in $H_2O_2$-mediated MAPK signaling in plants.

• Proceedings of the Korean Society of Plant Biotechnology Conference
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• 2003.04a
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• pp.65-71
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• 2003

Bax, a mammalian pro-apoptotic member of the Bcl-2 family, induces cell death when expressed in yeast. To investigate whether Bax expression can induce cell death in plant, we produced transgenic Arabidopsis plants that contained murine Bax cDNA under control of a glucocorticoid-inducible promoter. Transgenic plants treated with dexamethasone, a strong synthetic glucocorticoid, induced Bax accumulation and cell death, suggesting that some elements of cell death mechanism by Bax may be conserved among various organisms. Therefore, we developed novel yeast genetic system, and cloned several Plant Bax Inhibitors (PBIs). Here, we report the function of two PBIs in detail. PBI1 is ascorbate peroxidase (sAPX). Fluorescence method of dihydrorho-damine 123 oxidation revealed that expression of Bax in yeast cells generated reactive oxygen species (ROS), and which was greatly reduced by co-expression with sAPX. These results suggest that sAPX inhibits the generation of ROS by Bax, which in turn suppresses Baxinduced cell death in yeast. PBI2 encodes nucleoside diphosphate kinase (NDPK). ROS stress strongly induces the expression of the NDPK2 gene in Arabidopsis thaliana (AtNDPK2). Transgenic plants overexpressing AtNDPK2 have lower levels of ROS than wildtype plants. Mutants lacking AtNDPK2 had higher levels of ROS than wildtype. $H_2O_2$ treatment induced the phosphorylation of two endogenous proteins whose molecular weights suggested they are AtMPK3 and AtMPK6. In the absence of $H_2O_2$ treatment, phosphorylation of these proteins was slightly elevated in plants overexpressing AtNDPK2 but markedly decreased in the AtNDPK2 deletion mutant. Yeast two-hybrid and in vitro protein pull-down assays revealed that AtNDPK2 specifically interacts with AtMPK3 and AtMPK6. Furthermore, AtNDPK2 also enhances the MBP phosphorylation activity of AtMPK3 in vitro. Finally, constitutive overexpression of AtNDPK2 in Arabidopsis plants conferred an enhanced tolerance to multiple environmental stresses that elicit ROS accumulation in situ. Thus, AtNDPK2 appears to play a novel regulatory role in $H_2O_2$-mediated MAPK signaling in plants.

• Korean Journal of Soil Science and Fertilizer
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• v.35 no.5
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• pp.314-325
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• 2002

The effects of arbuscular mycorrhizal (AM) fungus (Glomus intraradices) on plant growth and N metabolic responses were examined in perennial ryegrass plants exposed to drought-stressed or well-watered condition. Mycorrhizal inoculation improved significantly leaf water potential, dry mass and P content. Drought stress increased significantly nitrate concentration in roots where the increase was much less in AM than non-AM. Drought stress decreased the concentration of soluble proteins in non-AM shoots, whereas non-significant decline occurred in AM shoots even under drought condition. The concentrations of ammonia and proline in drought stressed non-AM plants significantly increased, while mycorrhizal inoculation lowered significantly ammonia and proline accumulation. The decrease in leaf dry weight in drought stressed-plants was significantly correlated to the increase in ammonia (p<0.01) and proline concentration (p<0.01). These results suggested that the increased P content and N assimilation by mycorrhizal inoculation may be associated with drought stress tolerance, showing the moderating effects on shoot growth inhibition and ammonia accumulation in drought stressed-plants.

• Plant Biotechnology Reports
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• v.5 no.3
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• pp.225-234
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• 2011

Arabidopsis mutants with T-DNA insertion in seven calmodulin genes (CAM) were used to determine the specific role of CAM in the tolerance of plants to oxidative stress induced by paraquat and hydrogen peroxide ($H_2O_2$) treatments. Arabidopsis calmodulin mutants (cam) were screened for seedling growth, seed germination, induced oxidative damage, and levels of ${\gamma}$-aminobutyric acid (GABA) shunt metabolites. Only the cam5-4 and cam6-1 mutants exhibited an increased sensitivity to paraquat and $H_2O_2$ during seed germination and seedling growth. In response to treatments with $3{\mu}M$ paraquat and 1 mM $H_2O_2$, only the cam5-4, cam6-1 mutants showed significant changes in malonaldehyde (MDA) levels in root and shoot tissues, with highly increased levels of MDA. In terms of the GABA shunt metabolites, GABA was significantly elevated in root and shoot tissues in response to the paraquat treatments in comparison to alanine and glutamate, while the levels of all shunt metabolites increased in root tissue but not in the shoot tissue following the $H_2O_2$ treatments. GABA, alanine and glutamate levels were significantly increased in root and shoot of the cam1, cam4, cam5-4, and cam6-1 mutants in response to paraquat (0.5, 1 and $3{\mu}M$), while they were increased only in the root tissue of the cam1, cam4, cam5-4, and cam6-1 mutants in response to $H_2O_2$ (200 and $500{\mu}M$, 1 mM). These data show that the cam5-4 and cam6-1 mutants were sensitive to the induced oxidative stress treatments in terms of seed germination, seedling growth, and oxidative damage. The accumulation of GABA shunt metabolites as a consequence of the induced oxidative stress treatments (paraquat and $H_2O_2$ treatments) suggests that the GABA shunt pathway and the accumulation of GABA metabolites may contribute in antioxidant machinery associated with reactive oxygen species and in the acquisition of tolerance in response to induced oxidative stress in Arabidopsis seedlings.

• KSBB Journal
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• v.16 no.4
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• pp.344-350
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• 2001

The effect of salinity stress of Brassica olearacea and Capsicum annuum were studied at various levels of salinity conditions(Na-gluconate, K-gluconate, NaCl, KCl). The effects of salinity stress were measured by seedling growth rates and secondary metabolites contents of the stressed plants. Each seedling studied on the response of different salinity stress. Seedling growth of Capsicum annuum was inhibited up to 200 mM salt tolerance and Brassica olearacea was inhibited up to 400 mM salt tolerance. The produced anthocyanin was separated to high value from 200 mM NaCl in case of Brassica olearana and 50 mM K-gluconate in case of Capsicum annuum. The BADH activity was very high in Brassica olearacea seedlings treated with 200 mM NaCl and in Capsicum annuum seedlings treated with 100 mM K-gluconate. The BADH activities were increased during the early culture days, it induced betaine synthesis. The salinity stress promoted BADH activiy, subsequently endogenous betaine contents were increased, and it seemed to be secure seedling from salinity stress. The salinity concentration of 200 mM was effective on the inhibition of seed germination and on the increase of proline accumulation in tissue. The inhibition of seedling growth and accumulation of secondary metabolites in seedling were caused osmotic hypersensitivity against salinity stress.

• Journal of Environmental Health Sciences
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• v.20 no.2
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• pp.64-72
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• 1994

Tolerance to toxic effects of cadmium (Cd), including lethality has been shown following pretreatment with cadmium and zinc. This study was designed to determine if tolerance also develops to Cd-induced hepatotoxicity and renal toxicity. Three groups of rats (A, B, C), each consisting of 108 rats, were studied and each group was divided into three subgroups (1, 2, 3), 12 rats for each subgroup. Rats were subcutaneously pretreated with saline (A), CdCl$_2$ (0.5 mg/kg, B), and ZnCl$_2$ (13.0 mg/kg, C) during time periods of 5 days. At the end of the period, rats were challenged with CdCIa (3.0 and 6.0 mg/kg) by intraperitoneal injection. As for the cadmium levels in rat tissues after pretreatments, it was highest in the liver. Then kidney, heart, blood and muscle followed it in that order. After 24, 48 and 96 hours of intraperitoneal injection by challenge doses the concentration of cadmium in liver and kidney increased proportionally to the increase of challenge dosage. However metallothioneins in liver and kidney were increased by the pretreatment of cadmium and zinc. These data indicate the liver is a major target organ of acute Cd poisoning, and suggest that cadmium induced hepatic injury, via release of Cd-MT, may play and important role in the nephrotoxicity observed in response to short-term exposure to cadmium. This result suggests that increasing cadmium concentrations, gradually accumulating in liver and kidney as the result of the pretteatmerit, served to induced the synthesis of metallothionein, thus making them resistant to the challenge from cadmium.

• The Journal of Internal Korean Medicine
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• v.35 no.4
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• pp.428-438
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• 2014

Objectives: Obesity is an important cause of insulin resistance that leads to obese type 2 diabetes. Recently it has been found that obesity is associated with adipose tissue accumulation which causes systemic inflammation. In this study, we investigated effects of Inula helenium on the inflammation in high fat diet-induced insulin resistance mouse. Methods: Insulin resistance was induced in C57BL/6 male mice (19~21 g) on a 60% fat diet. Mice were divided into 3 groups (n=6) of normal, control and Inula helenium. After 12 weeks, body weight, FBS, oral glucose tolerance test (OGTT), serum level of insulin, epididymal fat pad, liver weight and the gene expression of tumor necrosis factor (TNF)-${\alpha}$, interleukin (IL)-6, interleukin (IL)-10 and cluster of differentiation (CD) 68 were measured. Also, adipose tissue macrophage was analyzed by fluorescence activated cell sorting. Results: Inula helenium significantly reduces oral glucose tolerance levels, insulin serum level and adipose tissue macrophage. Also Inula helenium increased IL-10 gene expression and decreased CD68 gene expression. Conclusions: These results show that Inula helenium has anti-insulin resistance and anti-inflammatory effects on a high fat diet-induced insulin resistance mouse model.

• Journal of Korean Medicine Rehabilitation
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• v.27 no.1
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• pp.1-17
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• 2017

Objectives This study aimed to investigate the effect of Bangkibokryeong-tang (BBT, Fangjifuling-tang) on blood glucose and body fat in high-fat diet-induced obese mice. Methods The experimental animals were divided into five groups- normal diet-fed control (ND), high-fat diet-fed control (HFD), HFD+BBT 75, HFD+BBT 150, and HFD+olistat as a positive drug control group. Markers of obesity, such as body weight, organ weight, diet efficiency, and serum levels of total cholesterol, triglycerides, lipid content, leptin, adiponectin, glutamic oxaloacetic transaminase (GOT)/glutamic pyruvic transferase (GPT)/lactate dehydrogenase (LDH), blood glucose, and insulin, were measured. Furthermore, results of the oral glucose tolerance test and ${\alpha}-glucosidase$ inhibition activity were examined in obese mice. Results Mice treated with BBT demonstrate lower body and organ weight, and reduced weight gain and food efficiency than that in the HFD-only control group. In addition, BBT decreased lipid accumulation in the liver and the levels of enzymes such as GOT, GPT, and LDH in the serum. Furthermore, the levels of triglycerides, total cholesterol, low density lipoprotein (LDL), and leptin were decreased in the serum but the levels of high density lipoprotein (HDL) and adiponectin were increased in the BBT-treated group compared with the control group. The BBT-treated group also demonstrated decreased blood glucose and insulin concentrations induced by feeding on a high-fat diet and improved glucose tolerance. Conclusions Based on the results above, BBT may reduce body fat and hyperglycemia in HFD-induced obesity. This suggests that BBT may be clinically useful in the treatment of obesity.

• Biomolecules & Therapeutics
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• v.26 no.6
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• pp.568-575
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• 2018

In order to discover lifespan-extending compounds made from natural resources, activity-guided fractionation of Zingiber officinale Roscoe (Zingiberaceae) ethanol extract was performed using the Caenorhabditis elegans (C. elegans) model system. The compound 6-gingerol was isolated from the most active ethyl acetate soluble fraction, and showed potent longevity-promoting activity. It also elevated the survival rate of worms against stressful environment including thermal, osmotic, and oxidative conditions. Additionally, 6-gingerol elevated the antioxidant enzyme activities of C. elegans, and showed a dose-depend reduction of intracellular reactive oxygen species (ROS) accumulation in worms. Further studies demonstrated that the increased stress tolerance of 6-gingerol-mediated worms could result from the promotion of stress resistance proteins such as heat shock protein (HSP-16.2) and superoxide dismutase (SOD-3). The lipofuscin levels in 6-gingerol treated intestinal worms were decreased in comparison to the control group. No significant 6-gingerol-related changes, including growth, food intake, reproduction, and movement were noted. These results suggest that 6-gingerol exerted longevity-promoting activities independently of these factors and could extend the human lifespan.