• Journal of fish pathology
• /
• v.32 no.2
• /
• pp.59-67
• /
• 2019

We developed and subsequently characterized mouse antibodies (MAbs) against viral hemorrhagic septicemia virus (VHSV, genotype IVa), the causative agent of VHS. Five hybridoma clones secreting MAbs against VHSV were established. The MAbs recognized the glycoprotein (MAbs 2C10, 18H4, 23H6, and 30B7) and nucleocapsid protein (15E10) of VHSV by western blot analysis. All five MAbs reacted with VHSV-infected cells and tissue homogenates of VHSV-infected olive flounder (Paralichthys olivaceus) by western blot analysis. Whereas, no reactivity was observed in normal cells and tissue homogenates of normal olive flounder. Moreover, these MAbs reacted with VHSV, but did not react with other fish viruses (infectious hematopoietic necrosis virus, hirame rhabdovirus, spring viraemia of carp virus, infectious pancreatic necrosis virus, marine birnavirus, and nervous necrosis virus) by enzyme linked immunosorbent assay (ELISA). These results indicate that the MAbs are specific to VHSV and can be of value in VHSV detection.

• Journal of fish pathology
• /
• v.10 no.2
• /
• pp.75-86
• /
• 1997

Solid phase enzyme linked immunosorbent assay (ELISA) was developed to detect the whole cells of Edwardsiella tarda from infected tissues of flounder. Cross-reaction test was performed by ELISA against fish pathogens such as A. hydrophila ATCC7966. V. anguillarum HYUFP5001, Y, ruckeri 11-4, E. ictaluri and Streptococcus sp. NG8206. Rabbit anti-E, tarda Edk-2 sera highly cross-reacted with A. hydrophila ATCC7966 and V. anguillarum HUFP5001. However, the cross-reaction was removed by using the anti-serum pre-adsorbed with A, hydrophila ATCC7966 FKC. The intra-species cross-reaction among E. tarda isolates was very high. ELISA with the whole cell antigens present in tissue homogenate appeared with highly decreased sensitivity, presumably by the co-coating of lipid or proteins in tissues. Thus, it would be necessary to use the infected tissue homogenates diluted more than 100 times with PBS for diagnosis. Interestingly, compared with the using of FKC antigen, the direct detection of viable cells in tissue homogenate showed more sensitive results with detection limit of $1{\times}10^3$ cells/ml in buffer or diluted tissue homogenate. Consequently, the ELISA method developed in this study was specific, rapid and sensitive for diagnosing edwardsiellosis.

• The Korean Journal of Physiology
• /
• v.4 no.2
• /
• pp.25-31
• /
• 1970

Tissue homogenates of Ehrlich ascites tumor tissues and several normal tissue of mice were incubated separately in medium maintaining $C^{14}$_acetate concentrations of 5, 10, 20, 30, 40, 50 and 60 mg%, in order to determine maximum oxidative rates of acetate. In every incubation experiments, respiratory $CO_2$ samples rapped by alkaline which was placed in the center well of the incubation blask were analyzed for total $CO_2$ Production rates and their radoactivies. The fractions of $CO_2$ from medium acetate to total $CO_2$ production rate were obtained with relative specific activities (RSA) which were calculated by ratio between specific activities (SA) of $CO_2$ and medium $CO^{14}$_acetate and $CO_2$ production rates from medium acetate were calculated from RSA and total $CO_2$ production rates. Maximum plateau values of oxidative rates described above were determined at incubation experiments of various concentrations of medium acetate and compared the oxidative rates of acetate of tumor with those of normal tissues such as kidney, brain and liver. Maximum plateau values of total $CO_{2}$ Production rates were obtained at acetate concentration of 20 mg% and represent $25.0{\pm}0.54\;{\mu}M/hr/gm$ in the brain, $16.3{\pm}2.5$ in the kidney, $9.1{\pm}1.78$ in the liver and $11.5{\pm}3.2\;{\mu}M/hr/gm$ in the ascites tuners. Substancial $CO_2$ yield was observed in the tumor tissues as in the normal tissues. On the other hand, plateau values of RSA were $25.7{\pm}1.04%$ in thee brain, $9.1{\pm}0.72%$ in the kidney, $2.5{\pm}0.73%$ in the liver and $0.51{\pm}0.12%$ in the tumor tissues. $CO_2$ yields from the medium acetate, were 4.19 in the kidney, 2.28 in the brain, 0.228 in the liter and $0.059\;{\mu}M/hr/gm$ in the tumor tissue. These show wide range even in the normal tissue but remarkable decrease in the tumor tissue. This fact means that further oxidation of acetate was inhibited remarkably in the tumor tissue.

• Analytical Science and Technology
• /
• v.30 no.3
• /
• pp.146-153
• /
• 2017

In this study, we isolated ginseng crude saponin (GCS) from Korean red ginseng (KRG) and determined the ginsenoside content in it to investigate the physiological and pathological effects of GCS on acute pulmonary inflammation induced by intratracheal instillation of cigarette smoke condensates (CSC) and lipopolysaccharide (LPS) solution in BALB/c mice. GCS was orally administered at doses of 10 mg/kg and 25 mg/kg for 3 weeks. The recovery rate of GCS from KRG was 6.5 % and total ginsenosides from GCS was 1.13 %, and the content of Rb1 was the highest among them. Total inflammatory cells in the lung homogenates and bronchoalveolar lavage fluid (BALF) increased following intratracheal administration of CSC and LPS. However, GCS administration impaired this increase. Furthermore, it inhibited the increase in leukocytes in the blood, considerably decreased neutrophils in BALF, and declined infiltration of inflammatory cells and deposition of collagen in the tracheal and alveolar tissue. In this study, GCS was found to have a protective effect against acute pulmonary inflammation and it may be beneficial in preventing various respiratory diseases.

• The Korean Journal of Physiology
• /
• v.9 no.1
• /
• pp.77-82
• /
• 1975

This experiment was carried out to observe a biological effect of ginseng on the weight gain, physical performance and lactic acid production after exercise in mice. A group of mice weighing about 19 gm was divided into ginseng and cotrol group. on the treadmill (Exp. I & II) and LDH activity of liver and heart homogenates (Exp. II) were determined. Results are summerized as follows; 1. Body weight gain was greater id ginseng group than in control and the difference was statistically significant at 9th and 16th days of experimental period. 2. Maximal running time of ginseng was found to be longer than that of control (p<0.05) in experiment I and the experiment II also revealed the significant increase in maximal running time in ginseng group. 3. Bloo lactate concentration of 48 hour-rest from physical exertion was lower in ginseng group than in control (p<0.05). 4. LDH activity in liver homogenate was lower compared to control group, but in heart homogenate, it was greater in ginseng group. Above findings may be concluded tat the range of biological dose (20 mg/mice/day) of ginseng powder stimulated the body weight gain and increase of physical performance and its mechanism might be attributable to a lower level of blood lactic acid. The adaptive change of LDH activity also contributed to the change in lactate level in blood and tissue.

• Archives of Pharmacal Research
• /
• v.20 no.1
• /
• pp.34-38
• /
• 1997

Recombinant human epidermal growth factor (rhEGF), a polypeptide of 53 amino acid residues, is subject to degradation by numerous enzymes, especially proteases, when it is applied on the skin for the treatment of open wound. Amastatin, aprotinin, bestatin, EDTA, EGTA, gabexate, gentamicin, leupeptin, and TPCK were investigated for the possible protease inhibitors, which may use to protect rhEGF from degradation by the enzymes in the skin. Skin homogenates containing protease inhibitors and rhEGF were incubated at $37^{\circ}C$ for 30 minutes. After the reaction was stopped with trifluoroacetic acid, the amount of rhEGF remaining in the sample was determined with an HPLC method. The percentages of rhEGF degraded, at the skin/PBS ratio of 0.25, in the mouse, rat, and human skin homogenate were 85%, 70%, and 46%, respectively. The degree of degradation of rhEGF in the cytosolic fraction was higher than that in the membrane fraction and these enzyme reactions were completed in 30 minutes. Bestatin, EGTA, and TPCK showed significant inhibitory effects on the degradation of rhEGF in the two fractions (p<0.05), while the other protease inhibitors had no significant inhibitory effects or, even resulted in deleterious effects. Therefore, the formulation containing one or several inhibitors among these effective inhibitors would be a promising topical preparation of rhEGF for the treatment of open wound.

• Archives of Pharmacal Research
• /
• v.21 no.2
• /
• pp.174-178
• /
• 1998

As a new colon-specific prodrug of 5-aminosalicylic acid (5-ASA), 5-aminosalicyl-glycine (5-ASA-Gly) was prepared by a simple synthetic route in good yield. Apparent partition coefficients of 5-ASA-Gly were lower than those of 5-ASA, which determined in$ CHCl_{3}$/pH 6.8 buffer or n-octanol/pH 6.8 buffer system. Stability of 5-ASA-Gly by peptidases was investigated by incubation of 5-ASA-Gly with the homogenates of tissue and contents of stomach, proximal small intestine or distal small intestine of rats at $37^{\circ}C$. 5-ASA was not detected, indicating that the prodrug was stable in the upper intestine. The amount of 5-ASA liberated from incubation of the prodrug in cecal or colonic contents of rats was about 65% or 27% in 8 hrs, respectively, which indicated that the prodrug activation took place more readily in the rat cecum whose bacterial counts are high like human colon. Results from in vitro experiments suggested 5-ASA-Gly as a promising candidate of a colon-specific prodrug of 5-ASA.

• Korean Journal of Veterinary Research
• /
• v.31 no.4
• /
• pp.471-477
• /
• 1991

Attempts to isolate chicken anemia agent (CAA) were made by inoculating tissue homogenates into MDCC-MSBl or LSCC-1104B1 cell lines and passaging the cells serially. CAA was isolated from the liver and thymus of 11 weeks old layer chickens and from the liver of 10 weeks old broiler breeder chickens. The layer flock experienced approximately 45% mortality during 9 to 14 week of age from gangrenous dermatitis and lymphoid organs of affected chickens were severely atrophied. The broiler breeder flock experienced approximately 7% mortality during 7 to 9 weeks of age and affected birds showed lesions of colibacillosis, staphylococcal arthritis, and coccidiosis together with atrophied lymphoid organs. The isolated viruses were identified as CAA by the indirect fluorescent antibody test and virus neutralization test using CAA immune sera including one to Gifu-1 strain of CAA. The CAA isolate 89-69, when inoculated into susceptible 1 day old SPF chicks, induced anemia 14 to 16 days after inoculation. It did not induce any cytopathic effects in chicken embryo liver and chicken embryo fibroblast cell cultures. Infectivity of the isolate was not affected by the treatment of chloroform or heat ($70^{\circ}C$ for 15 minutes).

• Journal of Marine Bioscience and Biotechnology
• /
• v.2 no.4
• /
• pp.263-266
• /
• 2007

Vibrio vulnificusis a causative agent of serious diseases in humans resulting from the contact of wound with seawater or consumption of raw seafood. Several studies aimed at detecting V. vulnificus have targeted vvh as a representative virulence toxin gene belonging to the bacterium. In this study, we targeted the rpoS gene, a general stress regulator, to detect V. vulnificus. PCR specificity was identified by amplification of 8 V. vulnificus templates and by the loss of a PCR product with 36 non-V. vulnificus strains. The PCR assay had the 273-bp fragment and the sensitivity of 10 pg DNA from V. vulnificus. SYBR Green I-based real-time PCR assay targeting the rpoS gene showed a melting temperature of approximately $84^{\circ}C$ for V. vulnificus strains. The minimum level of detection by real-time PCR was 2 pg of purified genomic DNA, or $10^3$ V. vulnificus cells from pure cultured broth and $10^3$ cells in 1g of oyster tissue homogenates. These data indicate that real-time PCR is a sensitive, species-specific, and rapid method for detecting this bacterium using the rpoS gene in pure cultures and in infected oyster tissues.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.51 no.4
• /
• pp.383-388
• /
• 2018

Goldfish hematopoietic necrosis (GHN) affects hematopoietic organs and causes high mortality in goldfish. The causative agent of GHN is cyprinid herpesvirus 2 (CyHV-2). The purpose of this study was to detect CyHV-2 in ornamental cyprinid fish. CyHV-2 monitoring was conducted on a monthly basis for 1 year using goldfish Carassius auratus and pearlscale goldfish Carassius auratus that had been cultured at a domestic aquafarm and imported from Singapore, respectively. The results of polymerase chain reaction (PCR) assays using specific primers for the CyHV-2 helicase gene showed that 27.8% of goldfish and 33.3% of pearlscale goldfish were positive for CyHV-2. No cytophatic effects were detected in koi fin or common carp brain cells inoculated with tissue homogenates from either fish. Our data provides the useful data for establishing future quarantine and disinfection policies.