Kim, Young-Gill;Choi, Min-Soon;Park, Sung-Woo;Lee, Keun-Kwang
Journal of fish pathology
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v.7
no.2
/
pp.161-171
/
1994
In order to elucidate the outbreak mechanisms of a new disease which is characterized by an intense congestion in central venus sinuses(CVS) of gill filaments in cultured eel. these experiments were carried out; epidemically surveyed on the cultured eel farms in the vicinity of Kunsan city and experimentaliy outbreaked the disease in the stressful condition such as thermal and handling shock and innoculated the supernatant from the homogenate of naturally severe congested gill into eels and onto the monolayer of the CHSE-214. Although the frequency of congestion in eels of B, C, D and E farms were higher than in eels of A farms, the water qualities(stocked and cultured water) among farms were not a great difference. In eels of B, C, D and E farms, the values of haematocrit(Ht), haemoglobin(Hb), total protein(Tp), albumin(Alb), glucose(Glu), magnesium(Mg) were lower and the values of calcium(Ca), methemoglobin(Met-Hb), glutamic pyruvic transminase(GPT), glutamic oxalacetic transminase(GOT) higher than in eels of A farms. These valules have not related to the frequency of congestion. An intensive congestion and dilataton in CVS of gill filaments in experimentally handling-stressed eels produced similar histopathological changes to those observed in the spontaneously diseased eel, but not in eels experimentally injected with filtering contents. The cytopathic effect on the CHSE-214 was not observed. In stressed eels the congestion of gill was increased in relation to either the decrease ranges of water temperature or the incerase in accllimated times. And increase in Ht, Met-Hb, Alb, Glu, GOT and GPT and decrease in Mg, Hb and Tp were found, which had a close relationship to congestion of gill. Cortisol were increased according to the decrease ranges in acclimated water temperature. From these results, decrease in water temperature during selection placed eels upon the stressed condition, made increase in ionic strength in blood stream, and CVS was dilatated owing to the increased blood inflow.
Kim, Je Hyeong;Yoon, Dae Wui;Jung, Ki Hwan;Kim, Hye Ok;Ha, Eun Sil;Lee, Kyoung Ju;Hur, Gyu Young;Lee, Sung Yong;Lee, Sang Yeub;Shin, Chol;Shim, Jae Jeong;In, Kwang Ho;Yoo, Se Hwa;Kang, Kyung Ho
Tuberculosis and Respiratory Diseases
/
v.67
no.2
/
pp.95-104
/
2009
Background: The pathophysiologic mechanisms of early acute lung injury (ALI) differ according to the type of primary insult. It is important to differentiate between direct and indirect pathophysiologic pathways, and this may influence the approach to treatment strategies. NF-$\kappa$B decoy oligodeoxynucleotide (ODN) is a useful tool for the blockade of the expression of NF-$\kappa$B-dependent proinflammatory mediators and has been reported to be effective in indirect ALI. The purpose of this study was to investigate the effect of NF-$\kappa$B decoy ODN in the lipopolysaccharide (LPS)-induced direct ALI model. Methods: Five-week-old specific pathogen-free male BALB/c mice were used for the experiment. In the preliminary studies, tumor necrosis factor (TNF)-$\alpha$, interleukine (IL)-6 and NF-$\kappa$B activity peaked at 6 hours after LPS administration. Myeloperoxidase (MPO) activity and ALI score were highest at 36 and 48 hours, respectively. Therefore, it was decided to measure each parameter at the time of its highest level. The study mice were randomly divided into three experimental groups: (1) control group which was administered 50 ${\mu}L$ of saline and treated with intratracheal administration of 200 ${\mu}L$ DW containing only hemagglutinating virus of Japan (HVJ) vector (n=24); (2) LPS group in which LPS-induced ALI mice were treated with intratracheal administration of 200 ${\mu}L$ DW containing only HVJ vector (n=24); (3) LPS+ODN group in which LPS-induced ALI mice were treated with intratracheal administration of 200 ${\mu}L$ DW containing 160 ${\mu}g$ of NF-$\kappa$B decoy ODN and HVJ vector (n=24). Each group was subdivided into four experimental subgroups: (1) tissue subgroup for histopathological examination for ALI at 48 hours (n=6); (2) 6-hour bronchoalveolar lavage (BAL) subgroup for measurement of TNF-$\alpha$ and IL-6 in BAL fluid (BALF) (n=6); (3) 36-hour BAL subgroup for MPO activity assays in BALF (n=6); and (4) tissue homogenate subgroup for measurement of NF-$\kappa$B activity in lung tissue homogenates at 6 hours (n=6). Results: NF-$\kappa$B decoy ODN treatment significantly decreased NF-$\kappa$B activity in lung tissues. However, it failed to improve the parameters of LPS-induced direct ALI, including the concentrations of tumor necrosis factor-$\alpha$ and interleukin-6 in BALF, myeloperoxidase activity in BALF and histopathologic changes measured by the ALI score. Conclusion: NF-$\kappa$B decoy ODN, which has been proven to be effective in indirect models, had no effect in the direct ALI model.
Park, Seungbae;Kang, Dong Hyeon;Jin, Changbae;Kim, Hyoung Ja
Journal of the Korean Society of Food Science and Nutrition
/
v.46
no.2
/
pp.210-219
/
2017
This study aimed to establish an optimal extraction process and high-performance liquid chromatography (HPLC)-photodiode array (PDA) analytical method for determination of marker compounds, dihydrokaempferol (DHK) and 3-O-methylquercetin (3-MeQ), as a part of materials standardization for the development of health functional foods from stems of Opuntia ficus-indica var. saboten (OFS). The quantitative determination method of marker compounds was optimized by HPLC analysis, and the correlation coefficient for the calibration curve showed very good linearity. The HPLC-PDA method was applied successfully to quantification of marker compounds in OFS after validation of the method in terms of linearity, accuracy, and precision. Ethanolic extracts from stems of O. ficus-indica var. saboten (OFSEs) were evaluated by reflux extraction at 70 and $80^{\circ}C$ with 50, 70, and 80% ethanol for 3, 4, 5, and 6 h. Among OFSEs, OFS70E at $80^{\circ}C$ showed the highest contents of DHK and 3-MeQ of $26.42{\pm}0.65$ and $3.88{\pm}0.29mg/OFS100g$, respectively. Furthermore, OFSEs were determined for their antioxidant activities by measuring 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging and lipid peroxidation (LPO) inhibitory activities in rat liver homogenate. OFS70E at $70^{\circ}C$ showed the most potent antioxidant activities with $IC_{50}$ values of $1.19{\pm}0.11$ and $0.89{\pm}0.09mg/mL$ in the DPPH radical scavenging and LPO inhibitory assays, respectively. To identify active components of OFS, various chromatographic separation of OFS70E led to isolation of 11 flavonoids: dihydrokaempferol, dihydroquercetin, 3-O-methylquercetin, quercetin, isorhamnetin 3-O-glucoside, isorhamnetin 3-O-galactoside, narcissin, kaempferol 7-O-glucoside, quercetin 3-O-galactoside, isorhamnetin, and kaempferol 3-O-rutinoside. The results suggest that standardization of DHK in OFSEs using HPLC-PDA analysis would be an acceptable method for the development of health functional foods.
Two experiments were conducted to study the nature of protein degradation in fish muscle postmortem, first one with English sole (Paraphyrus vetulus) followed by another with rockfish (Sebastodes spp.). In the first one, proteolysis was measured by the increase of amino-N in gutted fish during storage in ice and in the homogenates prepared from fish of different ice storage during $20^{\circ}C-incubation$. In order to test the possible involvement of fish muscle a cathepsin, a portion of each homogenate sample was exposed to 0.5 Mrad of gamma radiation to destroy viable microorganisms prior to the incubation. Proteolysis was not detected until viable count reached a level above $10^7$ cells per gm fish flesh, corresponding to 31 days of ice storage. Even if fish flesh were mechanically disrupted by means of homogenization and subsequently incubated at $20^{\circ}C$, proteloysis attributable to muscle cathepsin was not detected. In the second with rockfish muscle aseptically prepared from freshly killed fish, the samples were inoculated with a proteolytic strain of fish spoilage Pseudomonad or irradiated at 0, 0.5 and 3.0 Mrad. The four samle groups were stored at $0-2^{\circ}C$ to compare the spoilage pattern of sterile and non-sterile muscle. In sterile muscle both total-N (extracted in 0.5M KCl) and amino-N $(soluble\;in\;70\%\;ethanol)$ declined slightly while the inoculated muscle showing increase in parallel with the increase of number of inoculated bacterium. The results indicate that proteolysis is a part of normal fish spoilage and the onset of proteolysis is delayed until viable count reaches its maximum level. Contribution of fish muscle cathepsin to protein degradation in white flesh fish muscle post-mortem is nil.
We investigated the binding properties of $(^3H)$ QNB and $(^3H)$ NMS to mAchR to elucidate the characterstics of mAchR in rat brain by using two different preparations (homogemates & intact brain cell aggregates). The binding properties of both ligands demonstrated high affinity and saturability in both experiments, however $(^3H)$ QNB showed a significantly higher maximal binding capacity than tha ot $(^3H)$ NMS 1. In rat brain homogenates; Displacement of both lignands with several mAchR antagonists resulted in competition curves in accoradnce with the law of massaction for QNB, atropine & scopolamine in thie preparation, also a similar profile was found for the quaternary ammonium analogs of atropine & scopolamine (methyl atropine & methylscopolamine) when $(^3H)$ NMS was used to label the receptors in rat brain. But when these hydrophillic antagonists were used to displace $(^3H)$ QNB, they showed interaction with high- and low-affinity binding sites in brain homogenates. Pirenzepine, the nonclassical mAchR antagonist, was able to displace both ligands from binding sites in this preparation. 2. In intact rat brain cell aggregates; Intact bain cell aggregates were used to elucidate the binding characteristics of $(^3H)$ NMS to mAchR in rat. The magnitude of binding of this ligand was related linearly to the amount of cell protein in the binding assay with a high ratio of total to nonspecific binding. mAchR antagonists displaced specific $(^3H)$ NMS binding according to the law of mass-action, while it was possible to resolve displacement curves using mAchR agonist into high-& low-affinity component. 3. Our results indicate that more hydrophilic receptor ligand $(^3H)$ QNB, displacement experiments in both tissues demonstrated that the lipid solubility of a particulr mAchR ligand might play an important role in determining its profile of binding to the mAchR, and the concentrations of mAchR in rat brain are both on the cell surface (membrane-bound receptor) and in the intracelluar membrane (intermembrane-bound receptor). 4. The results are discussed in terms of the usefulness of dissociated intact rat brain cells in studying mAchR in central nervous system.
Kim, Min-Jung;Kim, Ji-Young;Leec, Seung-Jae;Yoon, Yong-Dal;Cho, Dong-Jae;Kim, Hae-Kwon
Development and Reproduction
/
v.5
no.1
/
pp.23-33
/
2001
When mammalian oocytes ovulate into the oviduct, associating follicular fluid components are exposed to the oviductal environment, possibly resulting in the mutual interaction between fillicu1ar and oviductal fluids. In the Present study, we have demonstrated for the first time that components of fallicular fluid could be modified by the oviductal fluid. Gelatin zymographic analyses of human follicular fluid (hFF) obtained from IVF patients showed consistently the presence of 110 kDa gelatinase (GA110) in addition to many bands among which 62 kDa gelatinase was predominant. Addition of EDTA or phenanfhroline to the gelatinase substrate buffer during gel incubation abolished GA110 band whereas phenylmethylsulffnyl fluoride (PMSF) did not. In contrast, bovine oviductal fluid(bOF) exhibited only 62 kDa gelatinase. Surprisingly, when bOF was added to hFF in 1:1 ratio and then the mixture was incubated for 3 h at 37$^{\circ}$C, GA110 of hFF disappeared. Disappearance of GA110 by bOF was observed even within 30 min after mixing with hFF. Addition of aminophenylmercuric acetate (APMA) to hFF also abolished enzymatic activity of GA110 but increased the activityof 62 kDa gelatinase. However, APMA abolished many other gelatinases as well unlike bOF. Interestingly, treatment of hFF with EDTA for 3 h remarkably increased the enzymatic activity of GA110 but not that of other gelatinases. Addition of phenanthroline, PMSF or soybean trypsin inhibitor (SBTI) did not affect overall gelatinase activities. Again, addition of bOF to the hFF pretreated with any of the above proteinase inhibitors abolished the appearance of GA110. Human serum also showed GAI 10 of which activity was greatlyenhanced by EDTA treatment. Similar to hFF, serum GA110 also disappeared by the addition of bOF. Human granulosa cell homogenate did not reveal any appreciable gelatinase activity except 92 kDa gelatinase. Anti-human gelatinase A antibody reacted with 62 kDa gelatinase of hFF. Based upon these results, it is concluded that bOF could selectively degrade an isoform of gelatinase A present in hFF and human serum.
Lipopolysaccharide(LPS) was posttreated after the 14 day-pretreatment of Ulva lactuca fractions(ULF), and their correlativity to enzymatic activity alteration was investigated in the liver of rats. ULF was intraperitoneally administered into rats at dose of $1m{\ell}/kg$ of 100 mg/kg concentration for 14 days. On the day 15, $1m{\ell}/kg$ of LPS was injected. The corelativity was examined by measuring the changed values of superoxide dismutase(SOD) in mitochondrial fraction and catalase(CAT), glutathione peroxidase(GPx) in liver homogenate. The results showed that LPS treatment decreased the high values of SOD, CAT, GPx to the low values, but ULF pretreatment increased the low values of SOD, CAT, GPx to the high values. It was suggested that ULF, LPS and antioxidative enzymes like SOD, CAT, GPx had the corelativity of the high-low-high pattern and that the ULF pretreatment played the proper preventive role in the protection against the LPS treatment-induced enzymatic inactivity in the water fraction.
The growth rate and heat resistance of two types of Bacillus cereus isolated from cooked rice were observed. The FB-1 strain showed positive to haemolysis and negative to starch hydrolysis, but FB-2 strain positive to both reaction. The cell number of B. cereus FB-1 reached to more than $10^7\;cells/ml$ within 6 to 12 hours at $25{\sim}35^{\circ}C$ when cultured on the medium of cooked rice homogenate (cooked rice 30g-phosphate buffer solution 80 ml), but the numbers at its maximum growth were only $10^4{\sim}10^6\;cells/ml$ at $45{\sim}55^{\circ}C$. The specific growth rate of FB-1 strain were $0.82hr^{-1}\;at\;20^{\circ}C$, $1.76hr^{-1}\;at\;30^{\circ}C$, $2.21hr^{-1}\;at\;35^{\circ}C$ and $1.84hr^{-1}\;at\;40^{\circ}C$, respectively. D-values of FB-1 and FB-2 spores at $70{\sim}100^{\circ}C$ were in the range from 18 to 3.1 min and 23.5 to 3.7 min, respectively.
Journal of the Korean Society of Food Science and Nutrition
/
v.32
no.1
/
pp.124-130
/
2003
Effect of Korean red ginseng (KRG) on the level of serum and liver lipids and lipid peroxidation was investigated in the rats fed high fat diet. Content of serum total cholesterol was significantly decreased (P<0.05) in KRG I group and KRG II group. Content of HDL-cholesterol was significantly increased by 69.75% and 39.15% in KRG I and KRG II group compared to control group, respectively. Atherogenic index (hi) was also significantly decreased by 74.76% and 37.38% in KRG I and KRG II groups compared to control group, respectively. Serum triglyceride content was significantly decreased (p<0.05) in only KRG II group. Antioxidative activity of KRG on the lipid peroxidation of serum and tissues in rats was also studied in vivo by measuring the formation of thiobarbituric acid reactive substances (TBARS). Contents of TBARS in the serum of both KRG groups were significantly decreased (p<0.05) and that of nonheme iron in serum was significantly increased (p<0.05) in a dose-dependent manner, which suggested that lipid peroxidation contents are inversely correlated with serum nonheme iron content. Content of TBARS in liver was significantly decreased (p<0.05) in KRG I and KRG II groups, without any influence in other tissues. Content of TBIARS in liver microsomal fractions stimulated by Fe$^{2+}$/ascorbate was significantly decreased (p<0.05) in KRG I and KRG II groups, whereas this observation did not occur in liver mitochondrial fractions. When the effect of KRG on TBARS content in the liver fractions of homogenates, microsomes, and mitochodria stimulated by Fe$^{2+}$/ascorbate was tested in vitro experimental model, TBARS of liver three fractions was significantly decreased at 6 mg/mL KRG compared with those of control. These results suggested that KRG powder have hypocholesterolemic effect as well as antioxidative effect in the serum and liver of the rats fed high fat diet.
The optimal pH of the extraction buffer was 7.5 considiering AChE stability and its buffer capacity when AChE was isolated and extracted from the housefly(Musca domesitca L.)and three other insect species with 0.01 M sodium phosphate buffer. Also, the optimal pH of the reaction buffer was 7.5 considering enzyme activity and its buffer capacity when AChE activity was measured with the substrate in 0.1 M sodium phosphate buffer. The Potter Elvehjem type homogenizer with Teflon pestle was used to homgenize the tissues. When preparing a AChE suspension by centrifuging the homogenate, 700 g supernatant of adult head for the housefly, 700 g supernatnat of 5th instar nymphal whole body for the brown planthopper, lipid-eliminated 10,000 g supernatant of 5th instar larval whole body for the diamondback moth, and 700 g supernatant of 4th instar larval head for the tobacco cutworm were considered satisfactory as enzyme sources in view of mass preparation, extraction efficiency and stability of enzyme activity during evaluation. When AChE suspensions of 4 insect species were stored at $-18^{\circ}C$, more than 90% of activity was maintained up to 3 weeks. Km values of AChEs of the housefly, the brown planthopper, and the diamondback moth were 0.042, 0.037 and 0.043 mM, respectively and AChE-specific substrate inhibition was observed at high concentration. Km value of the tobacco cutworm ChE was 1.15 mM and BuChE characteristics was observed, though further study is needed. The optimal substrate concentration for the AChE inhibition tests was 0.5 mM for the housfly, the brown planthopper, and the diamondback moth and 12 mM for the tobacco cutworm.
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