• Korean Journal of Veterinary Research
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• v.38 no.2
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• pp.273-279
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• 1998

The ability of bovine intact red blood cells to scavenge superoxide and hydrogen peroxide by superoxide dismutase, catalase and glutathione peroxidase was investigated. Intact red cells(up to 0.4%) suspensions did not inhibit ferricytochrome c reduction by superoxide in the superoxide generating system. On the other hand, intact red cell(0.4%) suspensions almost completely inhibit ferrocytochrome c oxidation by hydrogen peroxide. The ability of intact red cells to scavenge hydrogen peroxide was mainly attributed to either membrane bound catalase or glutathione peroxidase. The scavenge of hydrogen peroxide by 0.1~0.2% intact red cells showed a trend of dependence on mainly glutathione peroxidase. However, at blood cell concentration higher than 0.3%, the process depended upon peroxidase-independent scavengers like catalase. Enhancement of ferrocytochrome c oxidation by red cells treated with aminotriazole proved that the protection against hydrogen peroxide was due to catalase, while the protection in the presence of glutathione indicated scavenging effect of glutathione peroxidase against hydrogen peroxide.

• Journal of Environmental Health Sciences
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• v.22 no.2
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• pp.104-113
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• 1996

Cadmium uptake by growing and nongrowing (intact) cells of a chdmium-tolerant yeast Hansenula anomala B-7 in the presence of surfactants was studied. In growing cultures the addition of Triton X-100 or Tween 80 increased cadmium uptake by about 30% with no inhibition of cell growth, and in intact cells Triton X-100 increased cadmium accumulation by about 80% compared to surfactant-free controls. Considering balance between increased uptake and pollution, the addition of 0.1% Triton X-100 was preferable. By the mixed addition with defoamer silicone, during growth of cells Tween 80 or Triton X-100 enhanced uptake efficiency of cadmium compared to its single addition, whereas in intact biomass each of surfactants tested had no significant effect on cadmium uptake. The uptake of cadmium was observed to rise sharply to a maximum and then declined with increasing pH, and maximum accumulation of cadmium by growing and intact cells occurred at the pH of 6.0 and 7.0, respectively. A significant increase in cadmium uptake occurred with shaking culture. Cadmium uptake by growing and intact cells was almost completed during the culture time of 72 or 24 hrs, respectively. Scalded cells sorbed much more cadmium-ion than living cells.

• The Korean Journal of Mycology
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• v.19 no.4
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• pp.282-284
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• 1991

The yeast, Hansenula anomala B-7, isolated from the $Cd^{2+}$ rich soils and determined to be tolerant in the high concentration of $Cd^{2+}$ were employed in this work. Its intact cells grown in high concentration of $Cd^{2+}$ were observed to be Iysed at the early stage when transferred to a cadmium deficient broth. Its intact cells found to be not Iysed and grow well under the high concentration of $Cd^{2+}$. The Iysis of the intact cells grown at the high concentration of $Cd^{2+}$ ion was not found when the metal ions were replaced with $Cd^{2+}$ ion in the same concentration. This result indicated that Iysis of yeast cells, at least in this isolate, would be related to cell osmosis with the mineral ions added.

• Animal Bioscience
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• v.34 no.3_spc
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• pp.463-470
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• 2021

Objective: This experiment was conducted to find out the immunological effects of wheat phytase when long-chain inorganic polyphosphate (polyP) treated with wheat phytase was added to a macrophage cell line, Raw 264.7, when compared to intact long-chain polyP. Methods: Nitric oxide (NO) production of Raw 264.7 cells exposed to P700, a long-chain polyP with an average of 1,150 phosphate residues, treated with or without wheat phytase, was measured by Griess method. Phagocytosis assay of P700 treated with or without phytase in Raw 264.7 cells was investigated using neutral red uptake. The secretion of tumor necrosis factor α (TNF-α) by Raw 264.7 cells with wheat phytase-treated P700 compared to intact P700 was observed by using Mouse TNF-α enzyme-linked immunosorbent assay kit. Results: P700 treated with wheat phytase effectively increased NO production of Raw 264.7 cells by 172% when compared with intact P700 at 12 h exposure. At 5 mM of P700 concentration, wheat phytase promoted NO production of macrophages most strongly. P700, treated with wheat phytase, stimulated phagocytosis in macrophages at 12 h exposure by about 1.7-fold compared to intact P700. In addition, P700 treated with wheat phytase effectively increased in vitro phagocytic activity of Raw 264.7 cells at a concentration above 5 mM when compared to intact P700. P700 dephosphorylated by wheat phytase increased the release of TNF-α from Raw 264.7 cells by 143% over that from intact P700 after 6 h exposure. At the concentration of 50 μM P700, wheat phytase increased the secretion of cytokine, TNF-α, by 124% over that from intact P700. Conclusion: In animal husbandry, wheat phytase can mitigate the long-chain polyP causing damage by improving the immune capabilities of macrophages in the host. Thus, wheat phytase has potential as an immunological modulator and future feed additive for regulating immune responses caused by inflammation induced by long-chain polyP from bacterial infection.

• Animal Bioscience
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• v.36 no.10
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• pp.1604-1611
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• 2023

Objective: The aim of this study was to investigate the protective effect of wheat phytase as a structural decomposer of inflammatory nucleotides, extracellular adenosine triphosphate (ATP), and uridine diphosphate (UDP) on HT-29 cells. Methods: Phosphatase activities of wheat phytase against ATP and UDP was investigated in the presence or absence of inhibitors such as L-phenylalanine and L-homoarginine using a Pi Color Lock gold phosphate detection kit. Viability of HT-29 cells exposed to intact- or dephosphorylated-nucleotides was analyzed with an EZ-CYTOX kit. Secretion levels of pro-inflammatory cytokines (IL-6 and IL-8) in HT-29 cells exposed to substrate treated with or without wheat phytase were measured with enzyme-linked immunosorbent assay kits. Activation of caspase-3 in HT-29 cells treated with intact ATP or dephosphorylated-ATP was investigated using a colorimetric assay kit. Results: Wheat phytase dephosphorylated both nucleotides, ATP and UDP, in a dose-dependent manner. Regardless of the presence or absence of enzyme inhibitors (L-phenylalanine and L-homoarginine), wheat phytase dephosphorylated UDP. Only L-phenylalanine inhibited the dephosphorylation of ATP by wheat phytase. However, the level of inhibition was less than 10%. Wheat phytase significantly enhanced the viability of HT-29 cells against ATP- and UDP-induced cytotoxicity. Interleukin (IL)-8 released from HT-29 cells with nucleotides dephosphorylated by wheat phytase was higher than that released from HT-29 cells with intact nucleotides. Moreover, the release of IL-6 was strongly induced from HT-29 cells with UDP dephosphorylated by wheat phytase. HT-29 cells with ATP degraded by wheat phytase showed significantly (13%) lower activity of caspase-3 than HT-29 cells with intact ATP. Conclusion: Wheat phytase can be a candidate for veterinary medicine to prevent cell death in animals. In this context, wheat phytase beyond its nutritional aspects might be a novel and promising tool for promoting growth and function of intestinal epithelial cells under luminal ATP and UDP surge in the gut.

• Journal of Animal Science and Technology
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• v.63 no.1
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• pp.114-124
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• 2021

The objective of this study was to characterize the enzymatic hydrolysis of lipopolysaccharide (LPS) by wheat phytase and to investigate the effects of wheat phytase-treated LPS on in vitro toxicity, cell viability and release of a pro-inflammatory cytokine, interleukin (IL)-8 by target cells compared with the intact LPS. The phosphatase activity of wheat phytase towards LPS was investigated in the presence or absence of inhibitors such as L-phenylalanine and L-homoarginine. In vitro toxicity of LPS hydrolyzed with wheat phytase in comparison to intact LPS was assessed. Cell viability in human aortic endothelial (HAE) cells exposed to LPS treated with wheat phytase in comparison to intact LPS was measured. The release of IL-8 in human intestinal epithelial cell line, HT-29 cells applied to LPS treated with wheat phytase in comparison to intact LPS was assayed. Wheat phytase hydrolyzed LPS, resulting in a significant release of inorganic phosphate for 1 h (p < 0.05). Furthermore, the degradation of LPS by wheat phytase was nearly unaffected by the addition of L-phenylalanine, the inhibitor of tissue-specific alkaline phosphatase or L-homoarginine, the inhibitor of tissue-non-specific alkaline phosphatase. Wheat phytase effectively reduced the in vitro toxicity of LPS, resulting in a retention of 63% and 54% of its initial toxicity after 1-3 h of the enzyme reaction, respectively (p < 0.05). Intact LPS decreased the cell viability of HAE cells. However, LPS dephosphorylated by wheat phytase counteracted the inhibitory effect on cell viability. LPS treated with wheat phytase decreased IL-8 secretion from intestinal epithelial cell line, HT-29 cell to 14% (p < 0.05) when compared with intact LPS. In conclusion, wheat phytase is a potential therapeutic candidate and prophylactic agent for control of infections induced by pathogenic Gram-negative bacteria and associated LPS-mediated inflammatory diseases in animal husbandry.

• Journal of Plant Biology
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• v.36 no.4
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• pp.383-389
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• 1993

The effects of light and $CO_2$ on the electrophysiological characteristics of guard cells in the intact leaf and in the detached epidermis have been investigated. Guard cells in intact leaves showed the membrane hyperpolarization in response to light. The biggest induced change of the membrane potential difference (PD) in the guard cells of the intact leaf was 13 m V by light and 42 mV by $CO_2$ in Commelina communis. Similar results were obtained with Tradescantia virginiana. However, there were no changes of membrane PD in detached epidermis. In order to determine the influence of the mesophyll on the changes of membrane PD, infiltration of the mesophyll cells with photosynthetic inhibitors was performed. In CCCP infiltrated leaf discs the guard cell membrane was depolarized slightly by red-light and hyperpolarized by $CO_2$, but in leaf discs infiltrated with DCCD and DCMU the guard cell membrane was hyperpolrized by both red-light and $CO_2$ as the control leaf discs. In azide infiltrated leaf discs the guard cell membrane showed no response to light and there was a much reduced membrane hyperpolarization by $CO_2$ compared to other responses. It was likely that azide caused leaf damage and the activity of cell metabolism was decreased greatly, resulting in small membrane PD changes by $CO_2$ and no changes by redlight. Therefore, it can be suggested that red light was sensed by the mesophyll and the light induced guard cell membrane hyperpolarization was related to energy produced by cyclic-photophosphorylation, but ${CO_2}-induced$ guard cell membrane hyperpolarization was not related to photosynthesis. Alkalisation of the vacuole was observed when the intact leaf was exposed to $CO_2$, indicating that membrane hyperpolarization was mainly the result of proton efflux.efflux.

• Korean Journal of Environmental Biology
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• v.23 no.3 s.59
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• pp.221-227
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• 2005

The effects of light and $CO_2$ on the electrophysiological characteristics of guard cells in the intact leaf and isolated epidermis have been investigated. Fast hyperpolarization of guard cell apoplastic PD in the intact leaf was recorded reaching up to around 7 mV and 20 mV in response to light and $CO_2$. Whenever the experiments were attempted with isolated epidermis, there was no response to light and $CO_2$. In order to determine the influence of the mesophyll cells, the apoplastic PD of guard cells in isolated epidermis was measured in the presence of the mesophyll supernatant or the control medium. The apoplastic PD in isolated epidermis was hyperpolarized to -7mV, changing from -22mV to -29mV at 40 min. But, when isolated epidermis was incubated with the supernatant from mesophyll cells incubated in the light, the apoplastic PD in isolated epidermis was hyperpolarized to -19 mV, changing from -22 mV to -40.5 mV. $CO_2$ also caused a change of 0.1 to 0.3 pH unit in the intact leaf. However, this change was absent in isolated epidermis. A vibrating probe was used to detect the change in electrical currents at the surface of excised intact leaves and isolated epidermis. The reading of excised intact leaves in the dark was $0.5\muA\;cm^{-2},$ remaining steady until illuminated. Light increased the current on the surface of excised leaves to about $0.8\muA\;cm^{-2},$. However, light had no effect in the current on the surface of isolated epidermis. Apoplastic pH changes across the stomatal complex in response to light and dark were measured both in the intact leaves and isolated epidermis over the same time period using pH micro-electrodes. The guard cell wall of intact leaf was acidified to 2.5 pH unit, falling from pH 7.5 to pH 5.0 in the first 10 min. in the light. At the same time the guard cell wall pH of isolated epidermis fell from pH 7.5 to pH 7.0 at 10 min. The guard cell wall pH of isolated epidermis incubated in the mesophyll supernatant fell from pH 7.6 to pH 6.7 at 10 min. Likewise, It could be imagined that an electrical signal, chemicals and hormones propagated from the mesophyll in response to light and $CO_2$ could control a fast stomatal response.

• Journal of Microbiology and Biotechnology
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• v.3 no.4
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• pp.244-251
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• 1993

A novel system has been developed to produce ${\delta}-aminolevulinic$ acid (ALA) using the intact cells of late logarithmic phase of Rhodocyclus gelatinosus KUP-74. The system was shown optimum yield of extracellular ALA under a condition of anaerobic light irradiation (4 Klux) at $30^{\circ}C$ with no variation in cell mass. The rate of extracellular ALA formation was stimulated by low doses of either $C_4\;or\;C_5$ ALA biosynthetic precursors, where 5 mM ($C_4) and 3 mM ($C_5) of each precursors were appeared to generate the maximum rates of 3.3 and 4.0 nmoles of ALA per 0.35 mg cells per hr, respectively. Half-life of the system was 10 hr in a sense of an ability of portage transport of L-glutamate, and sequential dose of this compound was resulted in promising recovery of the ALA.

• Korean Journal of Animal Reproduction
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• v.14 no.2
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• pp.93-100
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• 1990

These experiments were carried out to investigate the effect of rabbit anti-bovine cumulus cell antibodies on in vitro maturation of bovine follicular oocytes. Antisera to bovine cumulus cell were produced Japanese Ginat rabbit by repeated immunization of intact or solubilized bovine cumulus cell and purified by ammonium sulfate precipitation and Sepharose CL-4B protein-A affinity chromatography. The bovine cumulus cell-specific antibodies were confirmed by indirect ELISA. The results obtained in these experiments were summarized as follows : 1. The titer of the antibodies to cumulus cell determined by indirect ELISA using intact or solubilized bovine cumulus cell coated plates was very high in both intact and solubilized cumulus cells. Namely, the optical density at 1:12,800 dilution of antibodies was still significantly higher than that of non-immunized control serum. 2. When the follicular oocytes were treated with antibody to intact cumulus cells, the maturation rate of cumulus compacted and removed oocytes was ranged 47.6 to 59.1%. These value is significantly lower(p<0.05) than that(78.8%) of follicular oocytes cultured without the antibody. 3. the maturation rate of cumulus compacted and removed oocytes treated with antibody to solubilized cumulus cells was ranged 46.7 to 59.1%, significantly lower(p<0.05) than that(82.1%) of ooyctes cultured in antibody free medium. From above mentioned results, it could be said that cumulus cells promote nuclear maturation of follicular oocytes and that the beneficial effect of cumulus cells to the oocyte maturation is inhibited by the action of antibody to cumulus cells.