• Journal of Aquaculture
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• v.15 no.1
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• pp.7-13
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• 2002

Since 1993, the White Spot Syndrome Virus (WSSV) disease occurred in China among cultured shrimps resulting in mass mortality. Epizootiological surveys undertaken during the outbreak period of 1993-1994 indicated that all stages of Penaeus chinensis, P. japonicus and P. monodon were infected. Consequent to the transport of contaminated shrimp seedlings and seawater, the disease spread all over the farms of China. The disease was more rapidly transmitted at temperatures above $25^{\circ}C$. Challenge experiments showed the causative agent was highly virulent. White spots appeared on the carapace of both span-taneous and experimentally infected shrimps. Moribund shrimps contained turbid hemolymph, hypertrophied Iymphoid organ and a necrotic mid-gut gland. Electron microscopy showed the presence of viral particles in the gills, stomach, lymphoid organ, and epidermal tissue of the infected shrimp. The visions were slightly ovoid with an envelope and averaged 350 $\times$ 150 nm; nucleocapsids measured 375 $\times$ 157 nm. With discontinuous sucrose gradient of 35, 50 and 60% (w/v), the virus was separated from hemolymph of the infected shrimp. The estimated molecular weight of genomic DNA was 237 Kb with EcoR I, 247 Kb with Hind III and 241kb with Pst I. A total of 9 hybridoma colones secreting monoclonal antibodies (MAbs) were produced from mouse myeloma and spleen cells immunized with WSSV. The immunofluorescence assay of gill tissue showed that the MAbs reacted with diseased but not with healthy shrimp. The MAbs belonged to IgGl, IgG2b subclass and IgM class, all with kappa light Immune-electron-microscopy with colloidal gold marker showed the presence of 5 MAbs epitopes on the envelope and one on the capsid of the virus. Baculoviral mid-gut gland necrosis showed the specificity of the MAbs produced. For diagnosis 5 different methods were selected. Using Kimura primers for PCR, or MAbs for immunoblot, ELISA or FAT method, in situ hybridization was carried out to show the gene. All these methods detected WSSV in the organ samples of the diseased shrimp but not in healthy one.

• Journal of Biomedical Engineering Research
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• v.14 no.4
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• pp.307-314
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• 1993

Plasma protein adsorption is the first event in the blood-material interaction and influenc- es subsequent platelet adhesion towards thlㅈombus formation. Thiㅈomboembolic events are strongly influenced by surface characteristics of materials and fluid dynamics inside the blood pump. In vitro flow visualizaion and an amimal experiment with the moving actuator type TAH were Performed in order to investigate fluid dynamic effects on the protein adsorption. The diffel'encl level, j of shear rate inside the ventricle Lvere determined by consid- ering the direction of the major opening of four healt valves in the implanted TAH and the visualized flow patterns as well. Each ventricle of the explanted TAH was sectionalized into 12 segments according to the shear rate level. The adsorbed protein on each segment was quantified using the ELISA method after soaking in 2% (wye)SDS/PBS for two days. Adsorbed protein layer thicknesses Itvere measured by the Immunogotd method under TEM. The SEM observation show that right ventricle (RV) , immobilized with albumin, displayed different degrees of platelet adhesion on each segment, whereas the left ventricle (LV), grafted by PEO-sulronate, indicated nearly , iame platelet adhesion behavior, regardless of shear rates. The surface concentrations of adsorbed proteins in the low shear rate region are hlghel'than those in the high region, which was confirmed statistically. A modified adsorption model of plasma protein onto polyurethane surface was suggested by considering the effect of the fluid dynamic characteristics.

• Asian Journal of Atmospheric Environment
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• v.6 no.1
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• pp.33-40
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• 2012

In this study, we characterized the physical form of allergenic Cry j 1 in the urban atmosphere. Through an immunofluorescence antibody method, we showed that allergenic Cry j 1 exists as fine particles (${\leq}1.1{\mu}m$). To determine Cry j 1 concentrations and its particle size distribution, we used the ELISA method to confirm that most Cry j 1 exists as fine particles in the urban atmosphere and is found at high concentrations on fine day next to rainy day. Furthermore, we evaluated Cry j 1 denaturation by using the Biacore J system based on the surface plasmon resonence (SPR) principle and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). We showed that the dissociation constant ($K_D$) of Cry j 1 that has been exposed to urban polluted air is lower ($1.76{\times}10^{-14}$ M) than that of Cry j 1 ($1.32{\times}10^{-9}-3.37{\times}10^{-9}$ M) of original pollen grains that has not been exposed to air pollutants. Cry j 1 turns into low molecular weight proteins by reacting with various acidic solutions. In sum, we showed that allergenic Cry j 1 exists as fine particles that can deposit in the lower respiratory tract. This finding clarifies the relationship between Japanese cedar pollinosis and air pollutants.

• Journal of Pharmacopuncture
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• v.11 no.1
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• pp.119-125
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• 2008

Objective : This experimental study was performed to investigate the effects of Houttuyniae Herba extract on anti-inflammation and anti-oxidation. Methods : The cytotoxicity of Houttuyniae Herba water extract and ethanol extract about viability of Raw 264.7 cell were tested using a colormetric tetrazolium assay(MTT assay). We investigated the inhibitory effects of Houttuyniae Herba water extract and ethanol extract on Propionibactrium acnes using paper disk diffusion method. To investigate the anti-inflammation effects of Houttuyniae Herba water extract and ethanol extract on LPS-induced macrophage Raw 264.7 cell, we used ELISA kit. We evaluated anti-oxidation effects of Houttuyniae Herba water extract and ethanol extract on HaCaT cell by Enzyme recycling method. Results : 1. In Houttuyniae Herba water extract and ethanol extract, cell toxicity depended on the density and wasn't difference between two extracts. 2. Houttuyniae Herba water extract and ethanol extract has not the significant inhibition effect of Propionibactrium acnes. 3. Concentration of 50, $100{\mu}g/m{\ell}$ Houttuyniae Herba water extract inhibited the production of NO in the Raw 264.7 cell stimulated with LPS. 4. All extracts except for $20{\mu}g/m{\ell}$ Houttuyniae Herba water extract showed anti-oxidation effect by decreasing the DPPH radicals. Conclusion : These results indicate that Houttuyniae Herba extract has anti-inflammation and anti-oxidation effects. If further study is performed, the use of Houttuyniae Herba extract will be valuable and benificial in the therapy of Propionibactrium acnes.

• IMMUNE NETWORK
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• v.3 no.4
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• pp.281-286
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• 2003

Background: Hepatitis B virus (HBV) infection is one of the worldwide public health problem affecting about 300 million people. The envelope protein of HBV consists of three components known as preS1, preS2, and S antigen. According to the recent study, anti-HBs Ab showed effective neutralization ability against HBV from chronic hepatitis B and liver transplant patients, suggesting the possible development of therapeutic antibody. Methods: Spleen cells immunized with S antigen of HBV were fused with myeloma cell line to obtain HBsAg specific monoclonal antibodies. High affinity antibodies against HBsAg (adr, ad and ay type) were selected by competitive ELISA method. Nucleotide sequence of the variable regions of monoclonal antibodies was analyzed by RT-PCR followed by conventional sequencing method. Results: We produced 14 murine monoclonal antibodies which recognize S antigen of HBV. Two of them, A9-11 and C6-9 showed the highest affinity. The sequence analysis of A9-11 revealed that variable regions of the heavy chain and light chains are members of mouse heavy chain I (B) and light chain lambda 1, respectively. Likewise, the sequence analysis of C6-9 revealed that variable regions of the heavy chain and light chains are members of mouse heavy chain II (B) and light chain kappa 1, respectively. Neutralization assay showed that A9-11 and C6-9 effectively neutralize the HBV infection. Conclusion: These results suggest that A9-11 and C6-9 mouse monoclonal antibodies can be used for the development of therapeutic antibody for HBV infection.

• Journal of the Korean Chemical Society
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• v.48 no.5
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• pp.467-472
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• 2004

Cyanobacterial toxins, microcystins which exist in korean lakes show strong toxicity to fish, cattles and human. In this study, we tried to degrade microcystin LR using various chemical oxidants, Chlorine, Potassium permanganate and Hydrogen Peroxide. The detection method for the concentrations of microcystin LR in water samples was Enzyme-Linked Immunosorbent Assay (ELISA) method using the monoclonal antibody of microcystin. Chlorine degraded microcystin LR effectively at the concentration of 800 pg/mL microcystin LR and 12 ppm chlorine. The reaction took 40 minutes at pH 7. Potassium Permanganate also degraded microcystin LR successfully at the concentration of 2000 pg/mL microcystin LR and 1.2 ppm chlorine. The degradation reaction took 60 minutes at pH 7. In the case of hydrogen peroxide, the degradation rate of microcystin LR was very slow because of the slow reaction rate.

• The Journal of Korean Medicine
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• v.24 no.2
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• pp.138-147
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• 2003

Objectives : In this study, we investigated the mechanism by which Chelidonium majus (CM) regulates nitric oxide (NO) production. Methods : Using mouse peritoneal macrophages, the mechanism by which CM regulates NO or tumor necrosis $factor-{\alpha}(TNF-{\alpha})$ production was examined. NO release was measured by the Griess method. $TNF-{\alpha}$ production was measured by the ELISA method. The protein extracts were prepared and samples were analyzed for the inducible NOS(iNOS) expression and nuclear factor kappa $B(NF-{\kappa}B)$ activation by Western blotting. Results : When CM was used in combination with recombinant $interferon-{\gamma}{\;}(rIFN-{\gamma})$, there was a marked cooperative induction of NO production. CM had an effect on NO production by itself. The expression of the iNOS gene was increased in $rIFN-{\gamma}$ plus CM-stimulated peritoneal macrophages and almost completely inhibited by pre-treatment with pyrrolidine dithiocarbamate (PDTC), an inhibitor of $NF-{\kappa}B$. The $NF-{\kappa}B$ activation was increased in rIFN-{\gamma} plus CM-induced peritoneal macrophages. The increased production of NO from $rIFN-{\gamma}$ plus CM-stimulated peritoneal rnacrophages was decreased by the treatment with $N^{G}-monomethyl-{_L}-arginine{\;}(N^{G}MMA){\;}N^{\alpha}-Tosyl-Phe$ chloromethyl ketone (TPCK) , and was almost completely inhibited by pre-treatment with PDTC. Furthermore, treatment with CM alone or rIFN-{\gamma} plus CM in peritoneal macrophages caused a significant increase in $TNF-{\alpha}$ production. PDTC decreased CM-induced $TNF-{\alpha}$ production significantly. After CM treatment in HT-29 or AGS cells, cell viability decreased. Conclusions : These findings demonstrate that CM increases the production of NO and $TNF-{\alpha}{\;}by{\;}rIFN-{\gamma}-primed$ macrophages and suggest that NF-B plays a critical role in mediating these effects of CM.

• The Korea Journal of Herbology
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• v.30 no.3
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• pp.69-76
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• 2015

Objectives : The effect of mixture extracted from Bupleuri Radix and Physalidis Herba(BR+RH) on the LPS-induced Depression in rats was investigated. Methods : Rats were administered intragastrically BR+PH after injectio of LPS to induce deprssion. Immobility was examined using Tail Suspension Test(TST), Forced Swimming Test(FST). The level of plasma corticosterone was measured by an Enzyme-Linked Immunosorbent Assay(ELISA) method. The expressions of c-Fos, Corticotropin Releasing Factor(CRF), NADPH-d in the Paraventricular nucleus(PVN) and TH in the Locus coeruleus(LC) were measured by immunohistochemical method. Results : In the effect of BR + PH on TST, immobility was significantly decreased comparing with the LPS group. In FST, immobility was shown decrease tendency in the BR+PH group. The expression of c-Fos in the PVN was significantly decreased at BR + PH400 group, comparing with the LPS group. The expression of CRF in PVN was shown dto have the decrease tendency in the BR+PH group, comparing with the LPS group. The expression of NADPH-d in PVN was not significantly decreased at BR+PH groups, comparing with the LPS group. The expression of TH in the LC was shown to have the decrease tendency at BR + RH groups, but not significantly, comparing with the LPS group. Conclusions : Anti-depressant effect of mixture after extracted from Bupleuri Radix and Physalidis Herba was through the anti-inflammatory effect via inhibition of HPA axis. NO and catecholamine system is not involved.

• Ocean and Polar Research
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• v.40 no.3
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• pp.169-176
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• 2018

Recently, the pop-up satellite archival tag (PSAT) is being used in studies as a method of using satellites for monitoring organisms. Because PSATs are attached directly on the organism being monitored, it is used mostly on larger species since the health of the organism is an important concern. For this reason, PSAT-based surveys are lacking in Korea with no studies on the physiological responses of organisms with PSAT attached. Accordingly, as a basic biomonitoring study using PSAT, the present study investigated the physiological changes in fish in response to the attachment of PSAT. The present study used red seabream (Pagrus major) and Korean rockfish (Sebastes schlegelii) as the experimental fish. The PSAT was attached to the muscle below the experimental fish's dorsal fin using a mono filament (n = 3). To investigate the changes in physiological responses according to PSAT attachment, blood samples were collected from all experimental fish, including the control (n = 3), at 1, 7, 14 and 21 days after the attachment. Upon blood sample collection, whole blood was used to measure hematocrit and hemoglobin levels. After separating the plasma, the separated plasma was used to measure the GOT, GPT, glucose, total protein, and cholesterol levels. Meanwhile, the plasma cortisol, superoxide dismutase, and catalase levels were measured using the ELISA method. The results showed that attaching the tag did not have any impact on the immunity and stress response of the experimental fish. The findings in this study also demonstrated the possibility of using PSAT for studying relatively smaller species living in the coastal waters of Korea.

• The Journal of Internal Korean Medicine
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• v.20 no.1
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• pp.122-132
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• 1999

Purpose : Hepatitis B virus DNA transfected cell line(HepG2.2.15) was cultured to evaluate the effect of herbs on the expression of HBeAg and the replication of HBV. HepG2.2.15 produces HBV particles as well as viral proteins into cell culture media. Methods : Extracts of herbs were adminitered to the cells on the proper concentration. Culture media was collected 48 hours after the herbal administration and HBeAg level in the media was examined by ELISA method. To confirm that the anti-viral effect was not due to direct cytotocixity of the extracts, normal cell proliferation was shown by cell counting. And as of the interference in protein synthesis of HepG2.2.15 by herb-extracts, we used the result of study that we performed before by ${\alpha}FP$ assay using EIA method. Results& Conclusion : Herb medicines like 地楡(Sanguisorbae Radix) and 覆盆子(Rubi Frusctus) showed significant inhibitory effect on HBeAg expression at p<0.01 and 五味子(Acanthopanacis Cortex) at p<0.05. Whereas, though some herbs such as ?草根(Rubiae Radix), 山査(Crataegii Fructus), 白芍藥(Paeoniae Radix Alba), and 大黃(Rhei Radix et Rhizoma) showed the tendecy to suppress HBeAg. most of them were not significant statistically. From the above, we could conclude that those herb medicines can be applied to patients effectively and further studies on effective fraction of some herbs are thought to be needed.