• IMMUNE NETWORK
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• v.3 no.2
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• pp.126-135
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• 2003

Background: One of the important factors in the prognosis of chronic hepatitis B patient is the degree of replication of hepatitis B virus (HBV). It has been known that HBV DNA polymerase plays the essential role in the replication of HBV. HBV DNA polymerase is composed of four domains, TP (Terminal protein), spacer, RT (Reverse transcriptase) and RNaseH. Among these domains, tyrosine, the $65^{th}$ residue of TP is an important residue in protein-priming reaction that initiates reverse transcription. If monoclonal antibody that recognizes around tyrosine residue were selected, it could be applied to further study of HBV replication. Methods: To produce TP-specific scFv (single-chain Fv) by phage display, mice were immunized using synthetic TP-peptide contains $57{\sim}80^{th}$ amino acid residues of TP domain. After isolation of mRNA of heavy-variable region ($V_H$) and light-chain variable region ($V_L$) from the spleen of the immunized mouse, DNA of $V_H$ and $V_L$ were obtained by RT-PCR and joined by a DNA linker encoding peptide (Gly4Ser)3 as a scFv DNA fragments. ScFv DNA fragments were cloned into a phagemid vector. scFv was expressed in E.coli TG1 as a fusion protein with E tag and phage gIII. To select the scFv that has specific affinity to TP-peptide from the phage-antibody library, we used two cycles of panning and colony lift assay. Results: The TP-peptide-specific scFv was isolated by selection process using TP-peptide as an antigen. Selected scFv had 30 kDa of protein size and its nucleotide sequences were analyzed. Indirect- and competitive-ELISA revealed that the selected scFv specifically recognized both TP-peptide and the HBV DNA polymerase. Conclusion: The scFv that recognizes the TP domain of the HBV DNA polymerase was isolated by phage display.

• Korean Journal of Environmental Agriculture
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• v.28 no.1
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• pp.59-68
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• 2009

Single-dose toxicity test of germanium-fortified lettuce was investigated in mice. Both sexes of C57BL/6 mice were orally administered once at a dose of 2,000 mg/kg. No death, clinical signs and pathological findings related to the treatment were observed. In addition, no significant changes in feed consumption and body weight gain were obtained during the treatment period, in spite of day-to-day fluctuation of water consumption. There were no considerable changes in hematology and serum biochemistry, except a significant decrease in GPT, GOT and LDH. Several alterations were observed in organ weight and blood biochemistry, including thymus, ovaries, heart, kidney and platelet in male or female mice. The ability of spleen cells proliferation was almost same level as shown in control group. However the population of B cells, helper T cells and cytotoxic T cells was not comparably changed in all groups. Taken together, it is suggested that single oral dose of germanium-fortified lettuce to C57BU6 mice did not cause apparent toxicological change at the dose of 2,000 mg/kg body weight.

• Mycobiology
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• v.40 no.3
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• pp.181-188
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• 2012

This study was initiated in order to investigate the anticancer and immunomodulating activities of crude polysaccharides extracted in methanol, neutral saline, and hot water (hereinafter referred to as Fr. MeOH, Fr. NaCl, and Fr. HW, respectively) from the fruiting bodies of Panellus serotinus. Content of ${\beta}$-glucan and protein in Fr. MeOH, Fr. NaCl, and Fr. HW extracts of P. serotinus ranged from 22.92~28.52 g/100 g and 3.24~3.68 g/100 g, respectively. In vitro cytotoxicity tests, none of the various fractions of crude polysaccharides were cytotoxic against sarcoma 180, HT-29, NIH3T3, and RAW 264.7 cell lines at the tested concentration. Intraperitoneal injection with crude polysaccharides resulted in a life prolongation effect of 23.53~44.71% in mice previously inoculated with sarcoma 180. Treatment with Fr. HW resulted in an increase in the numbers of spleen cells by 1.3 fold at the concentration of $50{\mu}g/mL$ compared with control. Treatment with Fr. NaCl resulted in improvement of the immuno-potentiating activity of B lymphocytes by increasing the alkaline phosphatase activity by 1.4 fold, compared with control, at the concentration of $200{\mu}g/mL$. Among the three fractions, maximum nitric oxide ($13.48{\mu}M$) was recorded at $500{\mu}g/mL$ in Fr. HW. Production of tumor necrosis factor alpha, interleukin-$1{\beta}$, and interleukin-6 was significantly higher, compared to the positive control, concanavalin A, at the tested concentration. Therefore, treatment with crude polysaccharides extracted from the fruiting body of P. serotinus could result in improvement of antitumor activity.

• Journal of the Korean Society of Food Science and Nutrition
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• v.34 no.10
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• pp.1509-1513
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• 2005

To develop a new functional food material, edible crude saponin was isolated from soybean cake using HP-20 resin and its anticancer effect and immune-activity were investigated. The saponin significantly inhibited the growth of cancer cells such as A549, MCF-7 and SW480 at a concentration of 1,000 $\mu$g/mL. Morphological changes was observed in the AS49 cells surface treated with the saponin of 1,000 $\mu$g/mL concentration. The proliferation of mouse spleen cells treated with saponin was increased in a dose-dependent manner compared with untreated control cells until the concentration of 1 $\mu$g/mL but decreased at higher concentrations than that. The NO production in marcrophage cell lines (RAW 264.7) treated with saponin was increased in a dosedependent manner compared with untreated control cells.

• Journal of the Korean Society of Food Science and Nutrition
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• v.44 no.5
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• pp.641-648
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• 2015

The purpose of this study was to determine the effects of dried Dendropanax morbifera leaf extracts on lipid profiles of mice fed a high-fat and -cholesterol diet (HFCD). ICR mice were divided into six groups based on mice fed AIN-93G diet (Normal), HFCD (Control), HFCD+100 mg/kg/d of D. morbifera leaf aqueous extract (DA-100), HFCD+200 mg/kg/d of D. morbifera leaf aqueous extract (DA-200), HFCD+100 mg/kg/d of D. morbifera leaf ethanol extract (DE-100), or HFCD+200 mg/kg/d of D. morbifera leaf ethanol extract (DE-200) for 7 weeks. The final body weights of mice fed D. morbifera extracts were all lower than those of the control group. Mice treated with D. morbifera extracts showed significantly reduced plasma and hepatic triglyceride (TG), total cholesterol (TC), and low-density lipoprotein cholesterol levels, along with increased plasma high-density lipoprotein cholesterol level. Fecal TG level was higher in DE-100 and DE-200 groups and TC level was significantly higher in the DA-200 and DE-200 groups. Relative liver weight, spleen weight, and testicle fat weight in mice treated with D. morbifera were reduced compared to the control group. Plasma insulin, aspartate transaminase, and alanine transaminase levels of experimental groups were also lower than those of the control group. All mice treated with D. morbifera extracts had lower malondialdehyde (MDA) content and higher superoxide dismutase (SOD) activity than the control group. Particularly MDA levels of the DA-200 and DE-200 groups and SOD levels of the DE-200 group were identical to levels of the normal group. These results suggest that D. morbifera extracts have lipid improvement effects in mice fed a HFCD.

• Korean Journal of Animal Reproduction
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• v.26 no.1
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• pp.25-32
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• 2002

This study were performed to investigate the effect of Bisphenol A(BPA) administration on semen characteristic and body or organ weight in mice. In BPA administration for 15 days or 30 days, sperm concentration and viability were lower in group treated with BPA than in control group. The proportion of sperm abnormality were significantly(P＜0.05) increased in groups administrated with BPA for 15 or 30 days than in control group. The administration of BPA in mice didn't affect the body and reproductive organ weight such as testis, epididymis, vasicular gland and coagulatin gland. The spleen weight were significantly affect, but liver and kidney weight were not affect in BPA administration groups for 15 days or 30 days This studies indicate that the short or long term BPA administration in mice were noxious effects on the semen characteristics and organ weight, but didn't affect body weight and reproductive organs.

• Journal of Microbiology and Biotechnology
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• v.15 no.3
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• pp.609-615
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• 2005

The safety and immunogenicity of an attenuated recombinant Salmonella vaccine strain, Salmonella enterica serovar Typhimurium llaB, was assessed. This vaccine strain could survive in low pH condition, and its ability of intracellular survival did not differ from that of S. enterica serovar Typhimurium UK1, which is the wild-type of the vaccine strain. The mortality of the mice orally administered with the vaccine strain was $50\%$ at the dose of $10^7$ CFU. All mice administered with $10^5\;or\;10^3$ CFU of the vaccine strain survived for 3 days postinoculation (pi). However, all mice administered with more than $10^3$ CFU of the vaccine strain died within 3 days pi. To examine the protective effect of the vaccine strain, mice were orally immunized with $10^4\;and\;10^6$ CFU of the bacteria. Control mice were given with 0.5 ml of phosphate buffered saline (PBS). After 8 days, the mice were challenged with $10^9$ CFU of S. enterica serovar Typhimurium UK1, and mortality was examined for 5 days. The survival rates of the mice immunized with $10^4\;and\;10^6$ CFU of the vaccine strain were $60\%\;and\;80\%$, respectively, whereas all control mice died within 2 days after challenging. To investigate the immunogenicity of S. enterica serovar Typhimurium llaB, mice were orally immunized with $10^5\;or\;10^6$ CFU ml of the vaccine strain. Five mice of each group were sacrificed at 5 and 12 days after immunization, and results showed that immunization of the vaccine strain led to increases of IgG1, IgG2, and IgM titers against S. enterica serovar Typhimurium UK1 in mouse sera, cytokine expressions such as IL-2, IL-4, IL-6, and IL-10 in spleen, and the lymphocyte proliferation response to mitogens (concanavalin A or LPS) stimulation.

• Animal cells and systems
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• v.1 no.1
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• pp.157-164
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• 1997

Ly-6E.1 antigen was proposed as a regulatory molecule of T lymphocyte activation, a hematopoietic stem cell marker, a memory cell marker, and an adhesion molecule. Though there were several reports suggesting the presence of Ly-6 ligand, the characterization of the ligand was not yet performed, As an attempt to screen the expression of Ly-6E.1 ligand, we prepared a probe for detecting Ly-6E.1 ligand by producing a fusion protein between Ly-6E.1 and $hlgC_{r1}$, A mammalian cell expression vector with Ly-6E.$1/hlgC_{r1}$ chimeric cDNA was transfected in SP2/0-Ag14 myeloma cells, and stable transfectants were selected. The fusion protein was produced as a dimer and maintained the epitopes for monoclonal antibodies specific for Ly-6E.1 and for anti-human lgG antibody. The purified fusion protein through Gammabind G column was used for FACS analyses for the expression of Ly-6E.1 ligand. The fusion protein interacted with several cell lines originating from B cells, T cells, or monocytes. The fusion Protein also strongly stained bone marrow, lymph node, and spleen cells, but thymic cells weakly, if any. The staining was more obvious in C57BL/6 $(Ly-6^b)$ than Balb/c $(Ly-6^a)$ mice. These results suggest that the interaction of Ly-6E.1 with Ly-6E.1 ligand may function both in the stem cell environment and in the activation of mature lymphocytes. The fusion protein may be a valuable tool in characterization of biochemical properties of the Ly-6E.1 ligand and, further, in isolating its cDNA.

• Journal of the Korean Society of Food Science and Nutrition
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• v.28 no.4
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• pp.917-923
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• 1999

To confirm therapeutic functionality of Urechis unicinctus which have been favored as a special seafood in Korea, the antitumor and immunological effect of those glycoprotein were studied. 4mg/kg dose of glycoprotein from Urechis unicinctus was most effective in solid tumor growth inhibition (43.63%) of sarcoma 180 cells. However, in case of mice injected with more than dose of 20mg/kg, tumor growth was not inhibited. The higher prolongation ratios were achieved at levels of 2mg/kg with 31.2% and 4mg/kg with 28.9%. The cytotoxic effect of glycoprotein on sarcoma 180 cells was increased slightly as administering level was increased. Number of total peritoneal exudate cells in all the glycoprotein administered groups increased remarkably meaning that Urechis unicinctus gly coprotein could help to improve immunity. Notable body weight change was not resulted in the glycoprotein treated mice compared with control group, but the ratios of both liver or spleen to body weight were increased in mice injected with 20mg/kg and 40mg/kg. These results suggest that the glycoprotein from Urechis unicinctus could stimulate immunity of the mouse bearing tumor cells. Furthermore, the number of leucocytes was also increased by 38.78% at the dose of 20mg/kg and by 46.30% of control at 40mg/kg, while the lower level of 2mg/kg or 4mg/kg showed no effect in increasing leucocyte number. The biochemical values such as GOT and GPT in serum were not changed in mice injected with glycoprotein in comparision with control group.

• Food Science and Preservation
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• v.14 no.6
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• pp.655-661
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• 2007

The biological activities of methanol extracts of Angelica gigas Nakai, such as antioxidation, anticancer and immuno-activity, were investigated in relation to development of functional foods. Anti-oxidation activity in the methanol extracts were assessed by hydrogen donating activity, reducing power and hydroxyl radical scavenging activity. Activities were dose-dependent over concentrations of 0.1, 0.5 and 1 mg/mL, with thehydrogen donating activity being over 50% at 1 mg/mL concentration. The methanol extracts inhibited the proliferation of SW480 cells in a dose-dependent manner, and chromatin condensation and apoptotic bodies were observed by fluorescence microcopy in the cells treated with the extracts for 24 hr. Caspase-3 activity was also increased in a dose-dependent manner in cells treated with the extracts relative to control cells. The extracts did not induce the proliferation of mouse spleen cells or NO production in macrophage cells (RAW 264.7). These results show that the methanol extract had slight anti-oxidative activity and did not increase immuno-activity, but inhibited proliferation of SW480 through apoptosis via a caspase dependent pathway.