• Biomolecules & Therapeutics
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• v.27 no.2
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• pp.193-200
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• 2019

Ceramide metabolism is known to be an essential etiology for various diseases, such as atopic dermatitis and Gaucher disease. Glucosylceramide synthase (GCS) is a key enzyme for the synthesis of glucosylceramide (GlcCer), which is a main ceramide metabolism pathway in mammalian cells. In this article, we developed a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method to determine GCS activity using synthetic non-natural sphingolipid C8-ceramide as a substrate. The reaction products, C8-GlcCer for GCS, could be separated on a C18 column by reverse-phase high-performance liquid chromatography (HPLC). Quantification was conducted using the multiple reaction monitoring (MRM) mode to monitor the precursor-to-product ion transitions of m/z $588.6{\rightarrow}264.4$ for C8-GlcCer at positive ionization mode. The calibration curve was established over the range of 0.625-160 ng/mL, and the correlation coefficient was larger than 0.999. This method was successfully applied to detect GCS in the human hepatocellular carcinoma cell line (HepG2 cells) and mouse peripheral blood mononuclear cells. We also evaluated the inhibition degree of a known GCS inhibitor 1-phenyl-2-decanoylamino-3-morpholino-1-propanol (PDMP) on GCS enzymatic activity and proved that this method could be successfully applied to GCS inhibitor screening of preventive and therapeutic drugs for ceramide metabolism diseases, such as atopic dermatitis and Gaucher disease.

• Fisheries and Aquatic Sciences
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• v.22 no.1
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• pp.1.1-1.8
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• 2019

Five experimental diets were formulated to evaluate the effects of dietary enzymatically hydrolyzed tuna by-product on growth, non-specific immune responses, and hematology of juvenile rainbow trout (Oncorhynchus mykiss). A basal diet with 50% of fishmeal was used as control (CON) and four other diets replaced 12.5% ($TBB_{12.5}$), 25% ($TBB_{25}$), 37.5% ($TBB_{37.5}$), and 50% ($TBB_{50}$) of fish meal in the CON diet. Juvenile rainbow trout ($4.87{\pm}0.05g$) were randomly distributed into 15 tanks (50 L) and fed 3-4% of wet body weight two times a day. At the end of 7 weeks of feeding trial, weight gain, specific growth rate, feed efficiency, and protein efficiency ratio of fish fed CON diet were significantly higher than those of fish fed $TB_{50}$ diet (P < 0.05). But there were no significant differences among fish fed CON, $TBB_{12.5}$, $TBB_{25}$, and $TBB_{37.5}$ diets (P > 0.05). There were no significant differences in GPT levels among fish fed CON, $TBB_{12.5}$, $TBB_{25}$, and $TBB_{37.5}$ diets. Also, there were no significant differences in lysozyme, superoxide dismutase, glucose, and total protein levels in all experimental diet (P > 0.05). The broken-line analysis indicated that the minimum dietary level of enzymatically hydrolyzed tuna by-product to replace fishmeal could be 29.7% in rainbow trout. These results indicated that the optimum level of dietary enzymatically hydrolyzed tuna by-product could replace greater than 29.7% but less than 37.5% of fishmeal in juvenile rainbow trout diet.

• 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 the Korean Crystal Growth and Crystal Technology
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• v.31 no.2
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• pp.78-83
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• 2021

In this study, we prepared NiCo2O4 nanoparticles with large surface area by hydrothermal synthesis. In order to optimize the processing conditions for spinel NiCo2O4 nanoparticles with large surface area, experimental variables including concentration of Ni and Co precursor, reaction time, and temperature for post-heat treatment were evaluated. Optimized conditions for spinel NiCo2O4 with large surface area were [Ni]/[Co] 1:2 ratio, reaction time for 12 h, and post-heat treatment at 400℃. To investigate the feasibility as potential application for glucose sensor, electrochemical tests of the prepared NiCo2O4 nanoparticles in response to glucose was performed, which suggests that the NiCo2O4 can be suitable for a non-enzymatic-based electrochemical glucose sensor based on its high sensitivity and selectivity for glucose detection.

• Journal of Marine Life Science
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• v.7 no.2
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• pp.94-101
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• 2022

In this study, the marine medaka Oryzias javanicus was exposed to two concentrations of non-toxin-producing red tide dinoflagellate C. polykrikoides (1,000 and 2,000 cells ml^-1) for 96 h, and the time-course biochemical responses of antioxidant and immunity parameters were analyzed in the liver tissue. Significant ichthyotoxicity with increasing cell concentrations of C. polykrikoides and exposure period was observed for 96 h. Opercular respiratory rate was lowered in marine medaka exposed to 2,000 cells ml^-1 of C. polykrikoides. Intracellular malondialdehyde (MDA) content significantly elevated in response to both cell concentrations. In the case of glutathione (GSH) content, the levels were significantly elevated by 1,000 cells ml^-1 of C. polykrikoides, but the contents significantly depleted upon exposure to 2,000 cells ml^-1 of C. polykrikoides. Similarly, enzymatic activities of catalase (CAT) and superoxide dismutase (SOD) were increased by 1,000 cells ml^-1 of C. polykrikoides, whereas their activities were lowered by 2,000 cells ml^-1 of C. polykrikoides. Analysis of the two immunity parameters, alternative complement pathway and lysozyme, showed significantly lowered activities in 2,000 cells ml^-1 of C. polykrikoides-exposed liver tissue. These biochemical effects of C. polykrikoides on marine medaka would be helpful for understanding its acute effects in marine fish.

• Proceedings of the Korean Society of Crop Science Conference
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• 2022.10a
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• pp.157-157
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• 2022

Plants being sessile are prone to various abiotic challenges, including salinity. Plants generally cope with salt stress by regulating their endogenous NO levels. NO exogenously applied in various forms also successfully impedes the salt stress, but its small size, short half life, and high volatility rate hamper its application in agriculture. NO application via CS as a nanocarrier is an alternate option to ensure the optimal kinetic release of NO for a long period compared to the free NO form. Herein, we synthesized and characterized GSNO-CS NP by ionic gelation of TPP with CS and then reacting with GSH, followed by reaction with NaNO₂ suspension. The synthesized NPs were characterized using non-destructive analytical techniques such as DLS, FTIR, and SEM to ensure their synthesis and surface morphology. NO-release profile confirmed optimal kinetic NO release for 24 h from NO-CS NP as compared to free NO form. The efficiency of NO-CS NP was checked on Arabidopsis plants under salinity stress by gauging the morphological, physiological, and enzymatic antioxidant system and SOS pathway gene expression levels. Overall, the results revealed that NO-CS NP successfully mitigates salinity stress compared to free GSNO. Concluding, the findings provide sufficient experimental evidence for the application of nanotechnology to enhance NO delivery, thus inducing more benefits for the plants under stress conditions by mitigating the deleterious impacts of salt stress on the morphological and physiological status of the plants, and regulating the ions exchange by overexpression of SOS pathway candidate genes.

• Journal of the Korean Society of Food Culture
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• v.38 no.6
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• pp.442-455
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• 2023

This study investigated the effects of pectinase treatment and skin contact time on the quality characteristics of Dae-hong peach wine. Wine was produced with variations in enzyme treatment and skin contact time (1 hour, 2 hours, 1 day, 2 days, and until the completion of fermentation). Enzyme treatment increased the production yield by 6%, as well as ethanol and redness levels, compared to the non-treated control. Volatile components were higher when the skin contact time was 2 hours or 1 day. Results were compared according to enzyme treatment and skin contact time and found to be influenced by methanol and 3-methyl-1-butanol. Enzyme treatment effectively enhanced yields and volatile compound contents. However, skin contact should be concluded a day before 1 day to ensure compliance with methanol legislative requirements. Therefore, our findings show that enzymatic treatment with shorter skin contact time preserves the distinctive characteristics of Dae-hong peaches and ensures the production of safe and flavorful wine.

• Journal of Ginseng Research
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• v.48 no.1
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• pp.1-11
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• 2024

Fresh ginseng is prone to spoilage due to its high moisture content. For long-term storage, most fresh ginsengs are dried to white ginseng (WG) or steamed for hours at high temperature/pressure and dried to form Korean Red ginseng (KRG). They are further processed for ginseng products when subjected to hot water extraction/concentration under pressure. These WG or KRG preparation processes affect ginsenoside compositions and also other ginseng components, probably during treatments like steaming and drying, to form diverse bioactive phospholipids. It is known that ginseng contains high amounts of gintonin lysophosphatidic acids (LPAs). LPAs are simple lipid-derived growth factors in animals and humans and act as exogenous ligands of six GTP-binding-protein coupled LPA receptor subtypes. LPAs play diverse roles ranging from brain development to hair growth in animals and humans. LPA-mediated signaling pathways involve various GTP-binding proteins to regulate downstream pathways like [Ca²⁺]_i transient induction. Recent studies have shown that gintonin exhibits anti-Alzheimer's disease and antiarthritis effects in vitro and in vivo mediated by gintonin LPAs, the active ingredients of gintonin, a ginseng-derived neurotrophin. However, little is known about how gintonin LPAs are formed in high amounts in ginseng compared to other herbs. This review introduces atypical or non-enzymatic pathways under the conversion of ginseng phospholipids into gintonin LPAs during steaming and extraction/concentration processes, which exert beneficial effects against degenerative diseases, including Alzheimer's disease and arthritis in animals and humans via LPA receptors.

• IMMUNE NETWORK
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• v.22 no.3
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• pp.26.1-26.18
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• 2022

IL-22, a pleiotropic cytokine, is known to have a profound effect on the regeneration of damaged intestinal barriers. The tissue-protective properties of IL-22 are expected to be potentially exploited in the attenuation and treatment of colitis. However, because of the disease-promoting role of IL-22 in chronic inflammation, a comprehensive evaluation is required to translate IL-22 into the clinical domain. Here, we present the effective production of soluble human IL-22 in bacteria to prove whether recombinant IL-22 has the ability to ameliorate colitis and inflammation. IL-22 was expressed in the form of a biologically active monomer and non-functional oligomers. Monomeric IL-22 (mIL-22) was highly purified through a series of 3 separate chromatographic methods and an enzymatic reaction. We reveal that the resulting mIL-22 is correctly folded and is able to phosphorylate STAT3 in HT-29 cells. Subsequently, we demonstrate that mIL-22 enables the attenuation of dextran sodium sulfate-induced acute colitis in mice, as well as the suppression of pro-inflammatory cytokine production. Collectively, our results suggest that the recombinant mIL-22 is suitable to study the biological roles of endogenous IL-22 in immune responses and can be developed as a biological agent associated with inflammatory disorders.

• Animal Bioscience
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• v.37 no.7
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• pp.1255-1262
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• 2024

Objective: The present study evaluated the effect of flaxseed meal degraded by a protease, Lactobacillus plantarum, or both on the growth performance, nutrient digestibility, and health status of broilers. Methods: There were four diets containing flaxseed meals in its non-degraded form (control, CON), degraded with 3,000 U/kg of protease (enzymatic, ELM), 1.0×10⁹ CFU/kg of Lactobacillus plantarum (fermented, FLM), or both (dual-degraded, DLM). Each form of flaxseed meals was added at 15% of diet. A total of 480 yellow-feathered broilers at 22 d of age were distributed into 4 groups with 6 replicates of 20 chickens each. The feeding trial lasted for 42 d. Growth performance, apparent fecal digestibility (dry matter, energy, crude protein, and ash), and serum immunoglobins and antioxidases were determined at 42 and 63 d of age. Results: Results showed that ELM, FLM, and DLM increased (p<0.001) the contents of peptides and decreased (p<0.001) cyanogenic glycosides, compared to CON. The diets with degraded flaxseed meals increased (p<0.05) feed intake and body weight gain throughout the feeding trial, and the digestibility of energy, crude protein, and ash at the end of feeding trial. Furthermore, all degraded groups enhanced (p<0.05) broiler health status by increasing serum immunoglobulins A and G. Additinally, DLM showed more pronounced effects (p<0.05) on these parameters than ELM or FLM. Conclusion: Flaxseed meals degraded by enzymolysis, fermentation, or both had improved nutrition and application in broilers.