The water and methanol extracts of Artemisia argyi showed significant cytotoxicities against J774A.1 cells but not so much against normal leukocytes. The cytotoxicities were found to be dependent on the extract concentration and the incubation time. The concentration of water and methanol extracts inhibiting 50% of cell proliferation ($IC_{50}$) were estimated to be 44.2 mg/ml and 71.6 mg/ml, respectively. In the presence of Artemisia argyi water extract, total superoxide dismutase (CuZnSOD and MnSOD) activities of media, cytoplasmic and mitochondrial fractions of J774A.1 cells increased in accordance with cytotoxicity. MnSOD was found to be the main component of enhanced total SOD activities, particulary in the mitochondrial fraction. In contrast to SOD, catalase and glutathione peroxidase (GPx) were not found in any instance of the current investigation. In addition, substantial amount of $O_2^-$ appeared to be generated in the mitochondrial fraction under the influence of Artemisia argyi. All data put together, it is postulated that Artemisia argyi extracts seem to stimulate $O_2^-$ generation in mitochondria of J774A.1 cells with concomitant increases of SODs. Since $H_2O_2$, the reaction product of SOD on $O_2^-$, is known to be readily converted to very toxic $OH{\cdot}$ in the absence of catalase and/or GPx cooperation, toxicity derived from ROS such as $O_2^-$, $H_2O_2$, and $OH{\cdot}$ may be the main cause of necrosis and/or apoptosis of J774A.1 cells.
To quantify the presence of mercuric ions in aqueous solution, double-stranded DNA (dsDNA) of poly(dT) was employed using a light switch compound, $Ru(phen)_2(dppz)^{2+}$ (1) which is reported to intercalate into dsDNA of a right-handed B-form. Addition of mercuric ions induced the dehybridization of poly(dT)${\cdot}$poly(dA) duplexes to form a hairpin structure of poly(dT) at room temperature and the metal-to-ligand charge transfer emission derived from the intercalation of 1 was reduced due to the dehybridization of dsDNA. As the concentration of $Hg^{2+}$ was increased, the emission of 1 progressively decreased. This label-free emission method had a detection limit of 0.2 nM. Other metal ions, such as $K^+$, $Ag^+$, $Ca^{2+}$, $Mg^{2+}$, $Zn^{2+}$, $Mn^{2+}$, $Co^{2+}$, $Ni^{2+}$, $Cu^{2+}$, $Cd^{2+}$, $Cr^{3+}$, $Fe^{3+}$, had no significant effect on reducing emission. This emission method can differentiate matched and mismatched poly(dT) sequences based on the emission intensity of dsDNA.
Polyphenol oxidase(PPO) was purified from an extract of Puerariae Radix by ammonium sulfate fractionation followed by Sephadex G-150 column chromatography, which resulted in a 56-fold increase in specific activity. The enzyme was optimum of pH 6.5. The optimum temperature of enzymic reaction was about $40^{\circ}$. The enzyme was thermostable with a half-life equal to 32 min at $70^{\circ}$. Km values of the PPO for catechol and pyrogallol from Lineweaver Burk plots were $1.3{\times}10^{-2}M$, $1.16{\times}10^{-2}M$, respectively. The substrate specificity of the Puerariae Radix PPO showed high affinity toward pyrogallol. Reducing reagents such as cysteine, potassium metabisulfite, ascorbic acid, 2-mercaptoethanol completely inhibited the PPO activity at $10^{-2}M$ level. Linewear-Burk analysis of inhibition data revealed that the inhibition by cysteine, 2-mercaptoethanol, 4-nitrocatechol, potassium cyanide was competitive with Ki values of $4.3{\times10^{-2}M,\;0.73{\times}10^{-6}M,\;6.9{\times}10^{-6}M,\;6.4{\times}10^{-7}M$, respectively. The browning reaction by PPO was observed to decrease temporarily with the addition of sodium diethyl dithiocarbamate, a well known copper chelating agent. Among the divalent cations, $Cu^{2+}$ ion was strong activator on PPO and $Mn^{2+},\;Co^{2+}$ ions was effect on PPO activity. $Zn^{2+},\;Mg^{2+}$ ions was inhibitor on PPO.
Seok, Hyeong Ju;Kim, Young Ryun;Kim, Tae Won;Hwang, Choul-Hee;Son, Min Ho;Choi, Ki-young;Kim, Chang-joon
Journal of the Korean Society of Marine Environment & Safety
/
v.28
no.1
/
pp.1-9
/
2022
The effect of sediments in a waste dumping area on marine organisms was evaluated using sediment toxicity tests with a benthic amphipod (Monocorophium acherusicum) and bioluminescent bacterium (Vibrio fischeri) in accordance with the Korean Standard Method for Marine Wastes (KSMMW). Nine sites in the East Sea-Byeong, East Sea-Jeong, and Yellow Sea-Byeong areas were sampled from 2016 to 2019. The test results showed that the relative average survival rate (benthic amphipods) and relative luminescence inhibition rate (luminescent bacteria) were below 30%, which were judged to be "non-toxic." However, in the t-test, a total of 12 benthic amphipod samples (6, 1, 1, and 4 in 2016, 2017, 2018, and 2019, respectively) were significantly different (p<0.05) from the control samples. To identify the source of toxicity on benthic amphipods, a simple linear regression analysis was performed between the levels of eight heavy metals (Cr, As, Ni, Cd, Cu, Pb, Zn, and Hg) in sediments and the relative average survival rate. The results indicated that Cr had the highest contribution to the toxicity of benthic amphipods (p = 0.000, R2 = 0.355). In addition, Cr was detected at the highest concentration at the DB-85 station and exceeded the Marine Environment Standards every year. Although the sediments were determined as "not toxic" according to the ecotoxicity criteria of the KSMMW, the results of the statistical significance tests and toxicity identification evaluation indicated that the toxic effect was not acceptable. Therefore, revising the criteria for determining the toxic effect by deriving a reference value through quantitative risk assessment using species sensitivity distribution curves is necessary in the future.
Kim, J.T.;Jang, H.Y.;Park, C.K.;Cheong, H.T.;Park, I.C.;Yang, B.K.
Asian-Australasian Journal of Animal Sciences
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v.24
no.2
/
pp.190-197
/
2011
The objective of the present study was to elucidate the fundamental mechanism of bovine oviduct epithelial cell (BOEC) co-culture on developmental capacity of bovine IVM/IVF embryos and to determine whether or not melatonin acts as an antioxidant in BOEC culture and subsequent embryo development. These studies examined the effects of melatonin against NO-induced oxidative stress on cell viability, lipid peroxidation (LPO) and the expression of antioxidant genes (CuZnSOD, MnSOD and Catalase) or apoptosis genes (Bcl-2, Caspase-3 and Bax) during BOECs culture. We also evaluated the developmental rates of bovine IVM/IVF embryos with BOEC co-culture, which were pre-treated with melatonin ($1,000\;{\mu}M$) in the presence or absence of sodium nitroprusside (SNP, $1,000\;{\mu}M$) for 24 h. Cell viability in BOECs treated with SNP (50-$2,000\;{\mu}M$) decreased while melatonin addition (1-$1,000\;{\mu}M$) increased viability in a dose-dependent manner. Cell viability in melatonin plus SNP ($1,000\;{\mu}M$) gradually recovered according to increasing melatonin addition (1-$1,000\;{\mu}M$). The LPO products were measured by thiobarbituric acid (TBA) reaction for malondialdehyde (MDA). Addition of melatonin in BOEC culture indicated a dose-dependent decrease of MDA, and in the SNP group among BOECs treated with SNP or melatonin plus SNP groups MDA was significantly increased compared with SNP plus melatonin groups (p<0.05). In expression of apoptosis or antioxidant genes detected by RT-PCR, Bcl-2 and antioxidant genes were detected in melatonin or melatonin plus SNP groups, while Caspase-3 and Bax genes were only found in the SNP group. When bovine IVM/IVF embryos were cultured for 6-7 days under the BOEC co-culture system pre-treated with melatonin in the presence or absence of SNP, the highest developmental ability to blastocysts was obtained in the $1,000\;{\mu}M$ melatonin group. These results suggest that melatonin has an anti-oxidative effect against NO-induced oxidative stress on cell viability of BOECs and on the developmental competence of bovine IVM/IVF embryo co-culture with BOEC.
The effect of different silicon (Si) sources and methods of application on the growth of two chrysanthemum cultivars grown in a soilless substrate was investigated. Rooted terminal cuttings of Dendranthema grandiflorum 'Lemmon Eye' and 'Pink Eye' were transplanted into pots containing a coir-based substrate. A nutrient solution containing 0 or $50mg{\cdot}L^{-1}$ Si from calcium silicate ($CaSiO_3$), potassium silicate ($K_2SiO_3$) or sodium silicate ($Na_2SiO_3$) was supplied once a day through an ebb-and-flood sub irrigation system. A foliar spray of 0 or $50mg{\cdot}L^{-1}$ Si was applied twice a week. Cultivar and application method had a significant effect on plant height. Cultivar, application method, and Si source had a significant effect on plant width. Of the three Si sources studied, $K_2SiO_3$ was found to be the best for the increasing number of flowers, followed by $CaSiO_3$ and $Na_2SiO_3$. In both the cultivars, sub irrigational supply of Si developed necrotic lesions in the older leaves at the beginning of the flowering stage as compared to the control and foliar spray of Si. Cultivar, application method, Si source, and their interactions had significant influence on leaf tissue concentrations of calcium (Ca), potassium (K), phosphorus (P), magnesium (Mg), sulfur (S), sodium (Na), boron (B), iron (Fe), and zinc (Zn). The addition of Si to the nutrient solution decreased leaf tissue concentrations of Ca, Mg, S, Na, B, Cu, Fe, and Mn in both cultivars. The greatest Si concentration in leaf tissue was found in 'Lemmon Eye' ($1420{\mu}g{\cdot}g^{-1}$) and 'Pink Eye' ($1683{\mu}g{\cdot}g^{-1}$) when $K_2SiO_3$ was applied through a sub irrigation system and by foliar spray, respectively.
Jang, Jae-Eun;Lim, Gab-June;Lee, Jin-Gu;Yoon, Seuong-Hwan;Hong, Sang Eun;Shin, Ki Hae;Kang, Chang-Sung;Hong, Sun-Seong
Journal of the Korea Organic Resources Recycling Association
/
v.27
no.2
/
pp.19-30
/
2019
Objective of this study was to develop an organic fertilizer utilizing domestic livestock horn meal and to investigate the application effect of rice and eggplant. The possibility of utilization of livestock horn meal as an organic resource to replace imported expeller cake fertilizer was examined. In order to select domestic organic resources with high nitrogen content, 8 kinds of organic matter such as chicken manure, fish meal, soybean meal, sesame meal, perilla meal, blood meal, livestock horn meal, and beer sludge were analyzed and organic resources with high nitrogen content were selected. In addition, the conditions for the production of organic fertilizers that can be used in organic agriculture were established by mixing of the rice husk biochar and the rice bran as the supplements with the raw materials for mixing ratios. The content of total nitrogen (T-N) in the livestock horn meal was 12.0 %, which was the next low in 13.5 % blood meal. The content of total nitrogen was 5.9 ~ 7.9 % in fish meal and oil cakes. Total nitrogen content of non-antibiotic chicken manure for organic farming was 3 % and nitrogen content in beer sludge was 3.5 %. Organic fertilizer was produced by using biochar, rice bran as a main ingredient of non-antibiotic chicken manure, livestock horn meal and beer sludge. Compared to nitrogen content (4.0 to 4.2 %) of imported expeller cake fertilizer (ECF), the nitrogen content of organic fertilizer utilizing domestic livestock horn meal is as high as 7.5 %. The developed organic fertilizer is met as Zn 400 mg/kg, Cu 120 mg/kg the quality of organic agricultural materials such as or less. To investigate the effect of fertilizer application on the crops, prototypes of developed organic fertilizer were used for the experiment under selected conditions. As a result of application the developed organic livestock horn meal fertilizer (LHMF) for cultivation of the rice and eggplant, the application quantity of the developed organic LHMF 100 % was decreased by 40 % compared to that of the mixed expeller cake fertilizer (MECF). The application of LHMF, which refers to the application rate corresponding to the nitrogen fertilization recommended by the soil test, was reduced by 40% compared to the application rate of MECF, but the same results were obtained in crop growth and yield. The selection of a new high concentration nitrogen source utilizing domestic organic resources and the development of organic fertilizer is the starting point of the research for substitution of imported ECF using domestic local resources at the present time that the spread of eco-friendly agriculture is becoming increasingly important. If it is expanded in the future, it is expected to contribute to the stable production of eco-friendly agricultural products.
Recently, NADPH oxidase 4 (NOX4)-mediated generation of intracellular reactive oxygen species (ROS) was proposed to accelerate adipogenesis of 3T3-L1 cell. We have previously shown that Cheonnyuncho (Opuntia humifusa) extract significantly inhibited adipocyte differentiation via downregulation of $PPAR{\gamma}$ (peroxisome proliferator-activated receptor gamma) gene expression. In this study, we focused on the molecular mechanism(s) of NOX4, G6PDH (glucose-6-phosphate dehydrogenase) and antioxidant enzymes in anti-oxidative activities of 3T3-L1 adipocytes. Our results indicate that Cheonnyuncho extracts markedly inhibits ROS production during adipogenesis in 3T3-L1 cells. Cheonnyuncho extracts suppressed the mRNA expression of the pro-oxidant enzyme such as NOX4 and the NADPH-producing G6PDH enzyme. In addition, treatment with Cheonnyuncho extract was found to upregulate mRNA levels of antioxidant enzymes such as Mn-SOD (manganese-superoxide dismutase), Cu/Zn-SOD (copper/zinc-SOD), glutathione peroxidase (GPx), glutathion reductase (GR), and catalase, all of which are important for endogenous antioxidant responses. These data suggest that Cheonnyuncho extract may be effective in preventing the rise of oxidative stress during adipocyte differentiation through mechanism(s) that involves direct down regulation of NOX4 and G6PDH gene expression or via upregulation of endogenous antioxidant responses.
Journal of Korean Society of Environmental Engineers
/
v.31
no.4
/
pp.241-248
/
2009
A farming area located near an abandoned copper mine in GuPo-ri, Choongchung province is heavily contaminated with heavy metals such as As, Pb, Cd, Cu and Zn of which concentrations are higher than the values typically detected in Korean soil environment. In this work, laboratory and field studies were conducted to examine feasibility of using Ca-citrate-phosphate solution in stabilizing heavy metals in the polluted soils. In laboratory batch experiments with field soil, the addition of Ca-citrate-phosphate solution resulted in decrease of aqueous phase concentration of phosphate and improvement of heavy metal stabilization, compared to those for sterilized soil samples. This indicates that microbial uptake of phosphate may have provided positive effects on availability of phosphate toward heavy metal stabilization. According to microbial community analysis for the field experiment, the use of Ca-citrate-phosphate led to increased diversity of microbial populations, and strict anaerobic microorganisms such as Anaerofilum and Treponema became the most dominant populations in the solution-amended field experiments. These findings suggest that, when Ca-citrate-phosphate is used for heavy metal stabilization in soils, microbial processes may have important roles in improving the stabilization of heavy metals by providing reducing conditions to the treatment locations or/and by making phosphate available to heavy metal stabilization.
Journal of Korean Society for Atmospheric Environment
/
v.21
no.1
/
pp.119-129
/
2005
The ambient TSP data measured at Seoul, Incheon. Taean, Daegu, Busan in Korea were used to explain the chemical composition and general features of Asian Dust (AD) observed in Korea. 9 episodes out of 19 were sampled from 2000 through May 2002, and measurements were conducted covering ionic and metal components with mass concentration. The results showed that daily averaged mass concentration (TSP) during the AD episodes was 458 $\mu\textrm{g}$/㎥, and ionic and metal concentrations were 27.93 $\mu\textrm{g}$/㎥ and 71.7 $\mu\textrm{g}$/㎥, respectively, accounting for 6.1 % and 15.5% of the total aerosol mass. TSP concentrations during episodes were varied from 120 to 1742 $\mu\textrm{g}$/㎥ according to the impact of Asian Dusts and had a tendency of showing higher values at sites in the west side of Korea, which can be explained by the effect of diffusion and deposition. In this study, ionic components like Ca (NO$_3$)$_2$, CaSO$_4$, NaNO$_3$, Na$_2$SO$_4$ were prominent types in secondary aerosol during AD periods and also indicated that V, Co as well as soil elements such as Ca, Fe, Mg, Mn, K correlated well with Al, while Cu, Cd, Pb, Zn didn't agree well with it. In addition, enrichment factors (EFs) for each metal component were obtained to provide simple information about source contribution of Asian Dust, and the results were compared with those from other AD studies. In this study, the results showed that aerosol properties in Korea during the Asian Dust were considerably different from those of general atmospheric condition and specially varied from case to case rather than site to site, which implies that there are certain variations in the soil of source region, pathways of air mass, and meteorological condition. For the enhanced study, those factors should be combined with the features of Asian Dust resolved from this study.
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