• Proceedings of the Korean Society of Applied Pharmacology
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• 1994.04a
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• pp.261-261
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• 1994

Bovine milk xanthine oxidase (E.C.1.1.3.22, XO) purchased from Sigma Chemical Co. had the three protein fragments below 150 kDa on 7.5％ SDS-PAGE, which did not show enzyme activity. To remove these fragments, the enzyme preparation was further purified through Sephadex G-200 column chromatography. Two peaks exhibiting enzymatic activity were separated very closely to the void volume, which were revealed as two different enzyme forms, dimeric and monomeric, confirmed by activity staining on native PAGE. Anti sera-against each of the two enzyme forms were raised by subcutaneous injection at multiple sites on the back of rabbits during 4 weeks. On the immunodiffusion test, it was found that both of the antisera of the two forms could react with each other, which implied that their epitopes were identical In the Western blot analysis of mouse liver cytosol fraction, it was found that rabbit anti-XO antibody bound well with the protein band of monomeric mouse liver XO of about 150kDa. Based on this result, mouse liver cDNA 1 ibrary was screened by in situ hybridizat ion wi th rabbi t anti -XO antibody as probe. Through the immunological screening, recombinant phages giving positive signal by the production of XO were selected and further purified. To validate these clones, purified phages were lysogenized in E. coli Y1089 and their lysates were analysed for enzyme activity and immunoreactivity, It was verified that lysates of the purified recombinant phage lysogens exhibited the enzymatic activity as well as bound wi th XO antibody, when induced by IPTG. The above results assert that selected recombinant phage carries mouse liver XO gene.

• Journal of Biomedical and Translational Research
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• v.19 no.4
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• pp.92-102
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• 2018

The objectives of this study were to find out the plant to enhance immune activity among 42 kinds of foods frequently consumed by the Korean elderly consisting of 5 food groups and 5 wild plants. Each sample was assessed the immunoactive effect by measuring $NF-{\kappa}B/AP1$ gene expression, nitric oxide and cytokine production in $RAW-Blue^{TM}$ cell. Soybean sprouts of 47 plants showed the highest $NF-{\kappa}B/AP1$ gene expression at the level of $1.13{\pm}0.03$ (O.D. 650 nm) and Soritae, sweet potato, banana, apple, garlic, crown daisy, cabbage and Ailanthus altissima also had high activity of $NF-{\kappa}B/AP1$ gene in $RAW-Blue^{TM}$ cell stimulated by LPS. NO production of Ailanthus altissima was significantly higher than that of other plants and 16 plants of glutinous sorghum, black rice, Seoritae, Heuktae, sweet potato, banana, apple, garlic, mungbean sprouts, spinach, crown daisy, young pumpkin, cabbage, soybean sprouts, Actinidia arguta and Aster scaber were the next best activity. The above results selected 17 out of 47 plant samples. Moreover, soybean sprouts was significantly shown to increase $TNF-{\alpha}$ ($1,509.55{\pm}1.38pg/mL$) and $IL-1{\beta}$ ($54.56{\pm}1.08pg/mL$) cytokines in comparison with RAW-Blue cell stimulated by LPS. According to the results of in vitro evaluation, the ethanol extract of soybean sprout increased the production of immune-enhancing cytokines by proliferation of macrophages. In addition, $NF-{\kappa}B$ transcription factor activity and NO production ability were excellent, and it was selected as a material having excellent immunological activity.

• The Korean Journal of Malacology
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• v.31 no.3
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• pp.187-194
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• 2015

This study was conducted to examine the effects of alterations in water temperature (WT) on biochemical and immunological factors in the hemolymph of the abalones, Haliotis gigantea and H. discus discus. The abalone were exposed to various WT; 18, 20, 22, 24, 26 and $28^{\circ}C$ for 96 hours. In biochemical factors, total-protein (TP), glucose, magnesium (Mg), aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were not significant changes in hemolymph of H. gigantea and H. discus discus. But calcium was significantly increased by high WT (${\geq}24^{\circ}C$). In immunological factor, The phenoloxidase (PO) activity was decreased in hemolymph of H. gigantea and H. discus discus exposed to high temperature (${\geq}22^{\circ}C$) compared to the control (P < 0.05). Whereas alkaline phosphatase (ALP) was not significantly changed. These results suggested that high temperature adversely affects the immunity of H. gigantea and H. discus discus.

• Korean Journal of Food Science and Technology
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• v.34 no.2
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• pp.325-329
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• 2002

This study was carried out to investigate the effect of porphyran on enzyme activity in rats and immunity in mice. Animals were divided into 5 groups, and were given porphyran diet for 4 weeks. Porphyran was extracted from Porphyra yezoensis: Diet groups were normal diet, control diet fed high fat, cholesterol and sodium cholate, control and 1% porphyran diet (1% PD), control and 5% porphyran diet (5% PD), control and 10% of porphyran diet (10% PD). Also Balb/c female mouse were injected i.p. with porphyran extract every other day for 20 days at levels of 1%, 2% and 5%. Alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP) and lactate dehydrogenase (LDH) activities were lower in the porphyran diet group than those in control group. Superoxide dismutase and catalase activities in liver homogenates were reduced in porphyran diet group compared to those of control group. Also, the level of liver thiobarbituric acid reactive substance (TBARS) was lower in porphyran group than that of control group. Porphyran increased IL-1 production in a dose-dependent manner, however, interleukine-2 production was reduced as the amount of porphyran increases. These results showed that supplementation of porphyran lowered antioxidant enzyme activities and has possibility of modulating immunological function.

• The Korean Journal of Malacology
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• v.31 no.1
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• pp.1-8
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• 2015

This study was conducted to investigate the effects of elevated water temperature (WT) on biochemical and immunological factors in the hemolymph of the abalones, Haliotis discus hannai and H. discus discus. The abalone were exposed to various WT; 20, 22, 24, 26 and $28^{\circ}C$ for 4 days. In the control and $20^{\circ}C$, total-protein (TP), glucose and calcium (Ca) in hemolymph of H. discus discus were higher than the values in H. discus hannai. The values of magnesium (Mg), alkaline phosphatase (ALP) and lysozyme in H. discus hannai were similar to the H. discus discus in the control. There were no significant alterations in TP, glucose and Mg levels of hemolymph in H. discus hannai and H. discus discus by WT increases. The values of Ca, ALP and lysozyme were increased in H. discus hannai exposed to the high temperature (26 and $28^{\circ}C$) compared to control, while the values in H. discus discus were not significant difference between the WT groups. The phenoloxidase (PO) activity was increased in hemolymph of H. discus hannai exposed to high temperature (${\geq}24^{\circ}C$) compared to the control (P < 0.05). These physiological and immunological parameters were significantly changed in H. discus hannai. However, these parameters in H. discus discus were barely altered at the high WT (P < 0.05). These results suggested that H. discus hannai is considered to be more sensitive than H. discus discus at the high WT.

• The Korean Journal of Mycology
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• v.39 no.3
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• pp.205-212
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• 2011

Phenylalanine ammonia-lyase (PAL) from the maize pathogen Ustilago maydis was analysed immunologically to obtain insights into the structural relationships between plant PAL and fungal PAL and between PAL and histidine ammonia-lyase (HAL). Cross-reactivity was found among all the PAL proteins from different species tested, using antibodies raised against both plant and fungal PALs. Both anti-Alfalfa and anti-popular PAL antibodies strongly recognized plant PALs but only weakly recognized fungal PALs. Antibodies raised against U. maydis PAL only weakly recognized the Rhodotorula glutinis yeast PAL. The anti-U. maydis PAL antibodies showed low affinity for the plant PALs but they bound strongly to Pseudomonas bacterial HAL. Significant cross-reactivity between the two plant PAL antibodies and the bacterial HAL was also observed. Both the anti-Ustilago PAL and the anti-poplar PAL antibodies displayed similar enzyme inhibition patterns, including moderate inhibition of bacterial HAL activity. However, the bacterial HAL antibody inhibited only Ustilago PAL. The PAL and HAL antibodies tested showed no inhibition against yeast PAL. This is first report on the immunological relationships between PAL and HAL.

• The Korean Journal of Physiology and Pharmacology
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• v.14 no.2
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• pp.71-75
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• 2010

To investigate the intestinal absorption of a fibrinolytic and proteolytic lumbrokinase extracted from Eisenia andrei, we used rat everted gut sacs and an in situ closed-loop recirculation method. We extracted lumbrokinase from Eisenia andrei, and then raised polyclonal antibody against lumbrokinase. Fibrinolytic activity and proteolytic activity in the serosal side of rat everted gut sacs incubated with lumbrokinase showed dose- and time-dependent patterns. Immunological results obtained by western blotting serosal side solution using rat everted gut sacs method showed that lumbrokinase proteins between 33.6 and 54.7 kDa are absorbed mostly by the intestinal epithelium. Furthermore, MALDI- TOF mass spectrometric analysis of plasma fractions obtained by in situ recirculation method confirmed that lumbrokinase F1 is absorbed into blood. These results support the notion that lumbrokinase can be absorbed from mucosal lumen into blood by oral administration.

• Korean Journal of Veterinary Research
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• v.54 no.4
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• pp.203-208
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• 2014

Bone marrow is a hematological and immunological organ that provides multiple immune cells, including B lymphocytes, and thus plays a critical role in the efficacy of vaccine. We previously demonstrated that Bordetella (B.) bronchiseptica antigen has high immunogenicity in spleen cells, a peripheral immune organ. In this study, we investigated the immunogenicity of B. bronchiseptica antigen in bone marrow cells, a central immune organ. B. bronchiseptica antigen increased the cellular activity of bone marrow cells and significantly enhanced the production of nitric oxide, IL-6, and TNF-${\alpha}$. Bone marrow cells primed with B. bronchiseptica antigen in vivo were harvested and stimulated with the same antigen in vitro. The stimulation of B. bronchiseptica antigen significantly increased the cellular activity and proliferation rate of the primed cells. B. bronchiseptica antigen also greatly induced the production of antigen-specific antibody in the primed cells. Taken together, the present study demonstrated that B. bronchiseptica antigen can stimulate bone marrow cells, a central immune organ, and recall the immune response of the primed bone marrow cells.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.9
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• pp.1438-1443
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• 2007

The experiment was conducted to evaluate the effects of Ligustrum lucidum (LL) and Schisandra chinensis (SC) on the growth, antioxidative metabolism and immunity of laying strain male chicks. The results showed that diets supplemented with 1% of either LL or SC had no effects on the growth performance of chicks compared with the control. Furthermore, both LL and SC significantly reduced malondialdehyde (MDA) concentration of serum and heart of chicks (p<0.05). In addition, superoxide dismutase (SOD) activity of serum of the birds was significantly elevated by supplementation with SC (p<0.05). Glutathione reductase (GR) activity of heart and serum of the birds was significantly elevated by supplementation with LL or SC (p<0.05). LL supplementation significantly elevated antibody values against Newcastle Disease virus (NDV)(p<0.05) and lymphoblastogenesis (p<0.05) of the birds. The results suggest that diets supplemented with 1% of either LL or SC may improve immune function and antioxidant status of chicks.

• BMB Reports
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• v.39 no.6
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• pp.671-676
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• 2006

The holoenzyme of protein phosphatase (PP) from tulip petals was purified by using hydrophobic interaction, anion exchange and microcystin affinity chromatography to analyze activity towards p-nitrophenyl phosphate (p-NPP). The catalytic subunit of PP was released from its endogenous regulatory subunits by ethanol precipitation and further purified. Both preparations were characterized by immunological and biochemical approaches to be PP2A. On SDS-PAGE, the final purified holoenzyme preparation showed three protein bands estimated at 38, 65, and 75 kDa while the free catalytic subunit preparation showed only the 38 kDa protein. In both preparations, the 38 kDa protein was identified immunologically as the catalytic subunit of PP2A by using a monoclonal antibody against the PP2A catalytic subunit. The final 623- and 748-fold purified holoenzyme and the free catalytic preparations, respectively, exhibited high sensitivity to inhibition by 1 nM okadaic acid when activity was measured with p-NPP. The holoenzyme displayed higher stimulation in the presence of ammonium sulfate than the free catalytic subunit did by protamine, thereby suggesting different enzymatic behaviors.