Journal of the Korean Society of Environmental Restoration Technology
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v.9
no.3
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pp.51-58
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2006
The purpose of this research presents a method to position and makes the structure for eco-corridors reasonably with collectable analysing results of various effects shown in mammals' road-kill at 429 points. Target animals of this research are Leopard cat, Siberian weasel, Raccoon dog, Korean hare, Eurasian red squirrel, Siberian chipmunk and Water deer. The results derived from the empirical analysis on the contents above are followed. First, according to the results as for Leopard cat road kill analysis, which is designated as Endangered Species Class II, the eco-corridor might be located at near village having stead food in order to decrease the frequencies of road-kill, because its road kill points were mainly collected at 4 lane hilly road with mountain-road-farm area geological type of. Second, because Siberian weasel's road kill was detected at 2 lane hilly road with mountain-road-stream geological type, the eco-corridor might be located at near a mill to decrease road-kill frequencies. Third, the road-kill frequency of Eurasian red squirrel can be reduced when the eco-corridor is located at the area across coniferous tree near 4 lane west sea freeway with mountain-road-mountain. Fourth, the road-kill of Raccoon dog can be reduced when the eco-corridor is located at 4 lane mountain road or hilly road with the geological type having farm land-road-mountain(stream). Fifth, Korean hare's road-kill can be reduced when the eco-corridor is located at grass land across ridge line of mountain, because wild rabbit road kill was happened at 4 lane mountain road or 2 lane mountain road(mountain-road-mountain). Sixth, As for Siberian chipmunk, the eco-corridor might be located at the side slope of mountain road at 2 lane mountain road under the speed of 60km/h with mountain-road-mountain. Seventh, For Water deer, the eco-corridor might be located at 4 lane hilly road with mountain-road-farm land. As for Common otter, Amur hedgehog, Yellow-throated marten, Weasel, it is difficult to specify the proper site of eco-corridor due to the lack of data. Eco-corridors for carnivores might be well located at 4 lane hilly road or 2 lane hilly road with mountain-road-farm land, and the track for herbivores might be well located as a overhead bridge on mountain-road-mountain type across mountains. In order to position eco-corridors for wildlife properly, we have to research animal's behavior with ecological background, and to consider the local uniqueness and regularly collect the empirical road-kill data in long term 3 to 5 year, which can be the foundation for the more suitable place of wild life eco-corridors.
In this study, the effects of replacing inorganic copper, zinc and manganese with different levels of organic complexes of the same trace minerals on the lipid peroxidation and antioxidant defense systems in broilers were investigated. Two-hundred Ross-308 one-day-old broiler chickens were placed on controlled diets until 42 d of age. The experimental animals were divided into four groups comprising three experimental groups and one control group, each consisting of 50 chickens. All groups were also divided into five subgroups each containing 10 broiler chicks. The mineral content of the control group diet was controlled using a standard inorganic mineral premix with supplement levels and sources of trace minerals typical of commercial broiler diets according to the National Research Council (NRC) (containing 8 mg Cu as $CuSO_4$, 40 mg Zn as $ZnSO_4$, and 60 mg Mn as MnO, per kg). In the experimental diets, mineral premix was also comprised of inorganic formulations, except for those of Cu, Zn and Mn. Organically-complexed Cu, Zn, and Mn were separately added to the basal diet at 1/3 (L1), 2/3 (L2) and 3/3 (L3) levels with respect to the NRC recommendation, as Bioplex $Cu^{TM}$, Bioplex $Zn^{TM}$, Bioplex $Mn^{TM}$. At the end of the trial, the plasma Zn level significantly increased when the plasma Cu level significantly decreased (p<0.05) in chickens fed at 2/3 and 3/3 levels of organically complexed minerals. The liver trace mineral concentrations were significantly higher in chickens fed inorganic trace minerals in comparison to those fed organically-complexed minerals. The plasma malondialdehyde (MDA) level of experimental chickens was decreased in groups receiving levels of organic Cu, Zn and Mn in comparison to those fed inorganic forms (p<0.01). The erythrocyte superoxide dismutase (SOD) activity was higher in all groups receiving the organic mineral supplements in comparison to those fed inorganic forms (p<0.01). No differences were observed on either the erythrocyte catalase (CAT) activity or the plasma ceruloplasmin (Cp) levels, and the liver MDA levels and liver CAT and SOD activities in any of the groups that received the organic supplements of Cu, Zn, and Mn. It was concluded that supplementation of lower levels of organically-complexed copper, zinc, and manganese instead of their inorganic forms in diets had no negative effects on the antioxidant defense system in broilers.
Arg13 is a conserved active-site residue in all known Pi class glutathione S-transferases (GSTs) and in most Alpha class GSTs. To evaluate its contribution to substrate binding and catalysis of this residue, three mutants (R13A, R13K, and R13L) were expressed in Escherichia coli and purified by GSH affinity chromatography. The substitutions of Arg13 significantly affected GSH-conjugation activity, while scarcely affecting glutathione peroxidase or steroid isomerase activities. Mutation of Arg13 into Ala largely reduced the GSH-conjugation activity by approximately 85 - 95%, whereas substitutions by Lys and Leu barely affected activity. These results suggest that, in the GSH-conjugation activity of hGST P1-1, the contribution of Arg13 toward catalytic activity is highly dependent on substrate specificities and the size of the side chain at position 13. From the kinetic parameters, introduction of larger side chains at position 13 results in stronger affinity (Leu > Lys, Arg > Ala) towards GSH. The substitutions of Arg13 with alanine and leucine significantly affected $k_{cat}$, whereas substitution with Lys was similar to that of the wild type, indicating the significance of a positively charged residue at position 13. From the plots of log ($k_{cat}/{K_m}^{CDNB}$) against pH, the $pK_a$ values of the thiol group of GSH bound in R13A, R13K, and R13L were estimated to be 1.8, 1.4, and 1.8 pK units higher than the $pK_a$ value of the wild-type enzyme, demonstrating the contribution of the Arg13 guanidinium group to the electrostatic field in the active site. From these results, we suggest that contribution of Arg13 in substrate binding is highly dependent on the nature of the electrophilic substrates, while in the catalytic mechanism, it stabilizes the GSH thiolate through hydrogen bonding.
Styrene is a commercially important chemical used mainly in the production of raw materials and plastics. To determine the effect of styrene on hepatic activities of antioxidant enzymes, styrene was treated to Sprague-Dawley rats at 50 mg/kg, 200 mg/kg and 400 mg/kg (i.p) twice a day for 4 days. There were determined the significantly increased activities of serum AST (aspartate aminotransferase), ALT (alanine aminotransferse), and the increased content of MDA (malondialdehyde) at the dose of 400 mg/kg compared to the control. The hepatic activities of XO (xanthine oxidase) and CYPdAH (cytochrome P450 dependant aniline oxidase) in the dose of 400 mg/kg compared to the dose of 200 mg/kg were more increased, which means the excessive ROS (reactive oxygen species)s were produced during Phase I. In addition, significantly decreased were rates of the hepatic activities of GPx (glutathione peroxidase), CAT (catalase), SOD (superoxide dismutase) and GST (glutathione S-transferase) at the dose of 400 mg/kg compared to the control. And, the group at the dose of 400 mg/kg showed more significantly decreased GSH (glutathione) level than the group at the dose of 200 mg/kg. The decrease in GSH could ascribe to the toxic metabolites of styrene, such as styrene oxide. In conclusion, these results indicate that the excessive ROSs and the toxic metabolites of styrene may result in the hepatotoxicity, and be related to their imbalanced activities for antioxidant enzymes.
Journal of Korean Society of Environmental Engineers
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v.33
no.6
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pp.396-404
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2011
An investigation on the removals of PAH-quinone compounds, which are commonly produced from the biological and/or chemical treatments of PAH-contaminated soils, from the aqueous phase via birnessite (${\delta}-MnO_2$)-mediated oxidative transformation is described. It was demonstrated that acenaphthenequinone (APQ), p-PAH quinone can be removed via birnessite-mediated oxidative-coupling reactions, and anthraquinone (AQ) and 1,4-naphthoquinone (1,4-NPQ), o-PAH quinones were efficiently removed by birnessite-mediated cross-coupling reactions in the presence of catechol (CAT) as a reactive mediator. The removals of PAH-quinone compounds followed pseudo-first-order reactions, and the rate constant (k, $hr^{-1}$) for the removals of 1,4-NPQ under the experiment conditions (1,4-NPQ = 10 mg/L, CAT = 50 mg/L, ${\delta}-MnO_2$ = 1.0 g/L, pH 5, Reaction time = 6~96 hr) was 0.0426, which was about 4 times lower than that of APQ (0.173). With the observed pseudo-first order rate constants with respect to birnessite loadings under the same experimental conditions, the surface-normalized specific rate constant, $K_{surf}$, for 1,4-NPQ was determined to be $8.5{\times}10^{-4}L/m^2{\cdot}hr$. The analysis of the kinetic data with respect to birnessite loading indicated that the cross-coupling reactions of 1,4-NPQ consist of two different reaction steps over time and the results have also been discussed in terms of the reaction mechanisms.
The purpose of this study was to examine the effect of Allium hookeri (AH) root on hepatic antioxidative enzyme contents in streptozotocin (STZ)-induced rats. Diabetes mellitus was induced in male Sprague-Dawley rats through injection of STZ dissolved in citrate buffer into tail veins at a dose of 45 mg/kg body weight. Sprague-Dawley rats were fed an AIN-93 recommended diet, and the experimental groups were fed a modified diet containing 5% and 10% of AH root powder for 4 weeks. The experimental groups were divided into four groups: a normal control (N-control), STZ-control, STZ-AH 5%, and STZ-AH 10% supplemented groups. The STZ-AH 5% group showed a significant increase in liver glycogen compared to the STZ-control group. Muscle glycogen and liver protein contents significantly increased in the AH-supplemented groups compared to the STZ-control group. The liver malondialdehyde content of the AH-supplemented group was significantly lower than that of the STZ-control group. Xanthine oxidase content was significantly reduced in all experimental groups. Glutathione-S-transferase content was significantly elevated in the AH-treated groups compared to the STZ-control group. Superoxide dismutase content was not significantly different among the experimental groups. Catalase content was significantly higher in the STZ-AH 10% group compared to the STZ-control group. These results show that supplementation with AH root may be useful for diabetic therapy and damage from oxidative stress.
Kim, Kee-Soon;Shin, Hong-Kee;Kim, Jin-Hyuk;Lee, Ae-Joo;Kang, Suck-Han
The Korean Journal of Physiology
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v.23
no.1
/
pp.151-167
/
1989
The present study was undertaken to investigate modification in electrophysiological characteristics of cat dorsal horn cells resulting from carrageenin-induced inflammation. The followings were studied; 1) the time-course of changes in responses of the WDR (wide dynamic range) cell 1-3h after subcutaneous injection of carrageenin in its receptive field; 2) the responses of the same dorsal hern cells before and after induction of inflammation; 3) the effect of inflammation on the responsiveness of dorsal horn neurons to algogens (bradykinin & potassium); and 4) the effect of inflammation on the activity of WDR cell following administration of indomethacin and clonidine. Though responses of WDR neuron were increased dramatically during first 1h, the maximal enhancement was observed 3h after induction of inflammation especially by repetitive light tactile stimulus. Following carrageenin injection the majority of WDR neurons (10/15 units) showed enhanced responses to all the mechanical stimuli while in 3 cases responsiveness were intensified during activation by one tactile stimulus (brush or pressure). One cell was unaffected by inflammation and in another case the response was enhanced only to noxious stimulus. Five of 9 cells that could initially be driven by noxious stimulus were activated more strongly by same stimulus and even by tactile stimulus (pressure) following inflammation. In 2 cases neurons were sensitized only to noxious stimulus whereas in another 2 cells that did not show enhanced responses to noxious stimulus responses to light tactile stimulus (pressure) appeared after inflammation. Of 16 LT cells tested 6 responded to squeeze while 4 showed the characteristics of WDR cell following inflammation. No modification in responsiveness was recognized in 3 cells whereas response to only brush was enhanced in another 3 neurons. Following carrageenin injection responses of LT cell to bradykinin or $K^{+}$ were not altered whereas those of WOR neurons to bradykinin or $K^{+}$ were suppressed in 22.2% and 33.3% of cases, respectively. In two of 8 activity of HT cells were inhibited by bradykinin while in five of 8 responsiveness to $K^{+}$ were rather enhanced by inflammation. In the rest inflammation was ineffective. In inflammation-induced animal the receptive field of LT cell was not changed whereas those of WDR cell and HT cell were tremendously expanded. The enhanced responses of WDR neurons to mechanical stimuli resulted from inflammation were suppressed by intravenously injected indomethacin and clonidine suggesting that postaglandin is involved in inflammation-induced sensitization of these cells. The involvement of peripheral and central mechanisms in the modification in responsiveness of dorsal horn cells in the carrageenin-induced inflammation was discussed.
Objective: Epigallocatechin-3-gallate (EGCG) is a major ingredient of catechin polyphenols and is considered one of the most promising bioactive compounds in green tea because of its strong antioxidant properties. However, the protective role of EGCG in bovine oocyte in vitro maturation (IVM) has not been investigated. Therefore, we aimed to study the effects of EGCG on IVM of bovine oocytes. Methods: Bovine oocytes were treated with different concentrations of EGCG (0, 25, 50, 100, and $200{\mu}M$), and the nuclear and cytoplasmic maturation, cumulus cell expansion, intracellular reactive oxygen species (ROS) levels, total antioxidant capacity, the early apoptosis and the developmental competence of in vitro fertilized embryos were measured. The mRNA abundances of antioxidant genes (nuclear factor erythriod-2 related factor 2 [NRF2], superoxide dismutase 1 [SOD1], catalase [CAT], and glutathione peroxidase 4 [GPX4]) in matured bovine oocytes were also quantified. Results: Nuclear maturation which is characterized by first polar body extrusion, and cytoplasmic maturation characterized by peripheral and cortical distribution of cortical granules and homogeneous mitochondrial distribution were significantly improved in the $50{\mu}M$ EGCG-treated group compared with the control group. Adding $50{\mu}M$ EGCG to the maturation medium significantly increased the cumulus cell expansion index and upregulated the mRNA levels of cumulus cell expansion-related genes (hyaluronan synthase 2, tumor necrosis factor alpha induced protein 6, pentraxin 3, and prostaglandin 2). Both the intracellular ROS level and the early apoptotic rate of matured oocytes were significantly decreased in the $50{\mu}M$ EGCG group, and the total antioxidant ability was markedly enhanced. Additionally, both the cleavage and blastocyst rates were significantly higher in the $50{\mu}M$ EGCG-treated oocytes after in vitro fertilization than in the control oocytes. The mRNA abundance of NRF2, SOD1, CAT, and GPX4 were significantly increased in the $50{\mu}M$ EGCG-treated oocytes. Conclusion: In conclusion, $50{\mu}M$ EGCG can improve the bovine oocyte maturation, and the protective role of EGCG may be correlated with its antioxidative property.
A comparative study of enzymatic properties between the trypsin from the cat-shark Cephaloscyllium umbratile ( C-T) and the two trypsins from the mackerel Scomber japonicus $(M-T_A\;and\;M-T_B)$ was carried out following after the previous paper(Pyeun et al., 1991). Trypsin from cat-shark(C-T) showed the higher heat stability compared to the others $(M-T_A\;and\;M-T_B)$ and its denaturation constant$(K_D)$ was $10.68\times10^{-4}\;sec^{-1}\;at\;55^{\circ}C$ with BA-p-NA substrate. The activation energies(Ea) of the trypsins measured at a temperature range from $30^{\circ}C\;to\;50^{\circ}C$ were estimated to be 4.07 kcal/mole for C-T, 11.61 kcal/mole for $M-T_A$, and 8.43kcal/mole for $M-T_B$, respectively. The Km values were $24.9\times10^{-5}\;M\;for\;C-T,\;5.37\times10^{-5}\;M\;for\;M-T_A,\;and\;9.65\times10^{-5}\;M\;for\;M-T_B$. On the other hand, the Ki values for TLCK and DFP determined by Dixon plot were $1.50\times10^{-6}\;M\;and\;9.28\times10^{-6}\;M\;for\;C-T\;2.86\times10^{-6}\;M\;and\;2.11\times10^{-4}\;M\;for\;M-T_A\;and\;3.90\times10^{-6}\;M\;and\;1.60\times10^{-4}\;M\;for\;M-T_B$ Similar amino acid profiles were showed between three trypsins each other, with few exceptions of $M-T_B$ containing higher amount of arginine, and the smaller amount of tryptophan in C-T than the others.
P450 1A2 is responsible for the metabolism of clinically important drugs and the metabolic activation of environmental chemicals. Genetic variations of P450 1A2 can influence its ability to perform these functions, and thus, this study aimed to characterize the functional significance of three P450 1A2 allelic variants containing nonsynonymous single nucleotide polymorphisms (P450 $1A2^*8$, R456H; $^*15$, P42R; $^*16$, R377Q). Variants containing these SNPs were constructed and the recombinant enzymes were expressed and purified in Escherichia coli. Only the P42R variant displayed the typical CO-binding spectrum indicating a P450 holoenzyme with an expression level of ~ 170 nmol per liter culture, but no P450 spectra were observed for the two other variants. Western blot analysis revealed that the level of expression for the P42R variant was lower than that of the wild type, however the expression of variants R456H and R377Q was not detected. Enzyme kinetic analyses indicated that the P42R mutation in P450 1A2 resulted in significant changes in catalytic activities. The P42R variant displayed an increased catalytic turnover numbers ($k_{cat}$) in both of methoxyresorufin O-demethylation and phenacetin O-deethylation. In the case of phenacetin O-deethylation analysis, the overall catalytic efficiency ($k_{cat}/K_m$) increased up to 2.5 fold with a slight increase of its $K_m$ value. This study indicated that the substitution P42R in the N-terminal proline-rich region of P450 contributed to the improvement of catalytic activity albeit the reduction of P450 structural stability or the decrease of substrate affinity. Characterization of these polymorphisms should be carefully examined in terms of the metabolism of many clinical drugs and environmental chemicals.
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