• Journal of Bio-Environment Control
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• v.32 no.1
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• pp.23-33
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• 2023

Sucrose (suc) is a disaccharide that consists of glucose (glu) and fructose (fru). It is a carbohydrate source that acts as a nutrient molecule and a molecular signal that regulates gene expression and alters metabolites. This study aimed to evaluate whether suc-specific signaling induces an increase in bioactive compounds by exogenous suc absorption via roots or whether other factors, such as osmotic stress or biotic stress, are involved. To compare the osmotic stress induced by suc treatment, 4-week-old cultured mugwort plants were subjected to Hoagland nutrient solution with 10 mM, 30 mM, and 50 mM of suc or mannitol (man) for 3 days. Shoot fresh weight in suc and man treatments was not significantly different from the control. Both man and suc treatments increased the content of bioactive compounds in mugwort, but they displayed different enhancement patterns compared to the suc treatments. Mugwort extract treated with suc 50 mM effectively protected HepG2 liver cells damaged by ethanol and t-BHP. To compare the biotic stress induced by suc treatment, 3-week-old mugwort plants were subjected to microorganism and/or suc 30 mM with Hoagland nutrient solution. Microorganisms and/or suc 30 mM treatments showed no difference about the shoot fresh weight. However, sugar content in mugwort treated with suc 30 mM and microorganism with suc 30 mM treatment was significantly higher than that of the control. Suc 30 mM and microorganism with suc 30 mM were effective in enhancing bioactive compounds than microorganism treatment. These results suggest that mugwort plants can absorb exogenous suc via roots and the enhancement of bioactive compounds by suc treatment may result not from osmotic stress or biotic stress because of microorganism, but by suc-specific signaling.

• Microbiology and Biotechnology Letters
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• v.51 no.1
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• pp.83-92
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• 2023

In this study, bioconversion of rutin to quercetin was confirmed by the fermentation of Korean indigenous probiotics and tartary buckwheat. Based on whole genome sequencing of 17 probiotics species, α-rhamnosidase, related to bioconversion of isoquercetin (quercetin 3-β-D glucoside) from rutin, is identified in the genome of CBT BG7, LC5, LR5, LP3, LA1, and LGA1. β-Glucosidase, related to bioconversion of isoquercetin to quercetin, is identified in the genome of all 17 species. Among the 17 probiotics species, 6 probiotics including CBT BG7, LR5, LP3, LA1, LGA1 and ST3 performed the bioconversion of rutin to quercetin up to 21.5 ± 0.3% at 7 days after fermentation. The fermentation of each probiotics together with enzyme complex Cellulase KN^® was conducted to reduce the time of bioconversion. As a result, CBT LA1 which showed the highest yield of bioconversion of 21.5 ± 0.3% when the enzyme complex was not added showed high bioconversion yield of 84.6 ± 0.5% with adding the enzyme complex at 1 day after fermentation. In particular, CBT ST3 (96.2 ± 0.4%), SL6 (90.1 ± 1.4%) and LP3 (90.0 ± 0.4%) showed high yield of bioconversion more than 90%. In addition, such probiotics including high levels in quercetin indicated the inhibitory effects of NO production in LPS-induced RAW264.7 cells. In this study, we confirmed that the fermentation of Korean indigenous probiotics and enzyme complex together with roasted tartary buckwheat increased the content of quercetin and reduced the time of bioconversion of rutin to quercetin which is a bioactive compound related to anti-inflammatory, antioxidants, anti-obesity, and anti-diabetes.

• Proceedings of the Korean Society of Near Infrared Spectroscopy Conference
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• 2001.06a
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• pp.4107-4107
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• 2001

Previous works have shown the viability of NIRS technology for the prediction of fatty acids in Iberian pig fat, but although the resulting equations showed high precision, in the predictions of new samples important fluctuations were detected, greater with the time passed from calibration development to NIRS analysis. This fact makes the use of NIRS calibrations in routine analysis difficult. Moreover, this problem only appears in products like fat, that show spectrums with very defined absorption peaks at some wavelengths. This circumstance causes a high sensibility to small changes of the instrument, which are not perceived with the normal checks. To avoid these inconveniences, the software WinISI 1.04 has a mathematic algorithm that consist of create a “Repeatability File”. This file is used during calibration development to minimize the variation sources that can affect the NIRS predictions. The objective of the current work is the evaluation of the use of a repeatability file in quantitative NIRS analysis of Iberian pig fat. A total of 188 samples of Iberian pig fat, produced by COVAP, were used. NIR data were recorded using a FOSS NIRSystems 6500 I spectrophotometer equipped with a spinning module. Samples were analysed by folded transmission, using two sample cells of 0.1mm pathlength and gold surface. High accuracy calibration equations were obtained, without and with repeatability file, to determine the content of six fatty acids: miristic (SECV$\sub$without/=0.07% r$^2$$\sub$without/=0.76 and SECV$\sub$with/=0.08% r$^2$$\sub$with/=0.65), Palmitic (SECV$\sub$without/=0.28 r$^2$$\sub$without/=0.97 and SECV$\sub$with/=0.24% r$^2$$\sub$with/=0.98), palmitoleic (SECV$\sub$without/=0.08 r$^2$$\sub$without/=0.94 and SECV$\sub$with/=0.09% r$^2$$\sub$with/=0.92), Stearic (SECV$\sub$without/=0.27 r$^2$$\sub$without/=0.97 and SECV$\sub$with/=0.29% r$^2$$\sub$with/=0.96), oleic (SECV$\sub$without/=0.20 r$^2$$\sub$without/=0.99 and SECV$\sub$with/=0.20% r$^2$$\sub$with/=0.99) and linoleic (SECV$\sub$without/=0.16 r$^2$$\sub$without/=0.98 and SECV$\sub$with/=0.16% r$^2$$\sub$with/=0.98). The use of a repeatability file like a tool to reduce the variation sources that can disturbed the prediction accuracy was very effective. Although in calibration results the differences are negligible, the effect caused by the repeatability file is appreciated mainly when are predicted new samples that are not in the calibration set and whose spectrum were recorded a long time after the equation development. In this case, bias values corresponding to fatty acids predictions were lower when the repeatability file was used: miristic (bias$\sub$without/=-0.05 and bias$\sub$with/=-0.04), Palmitic (bias$\sub$without/=-0.42 and bias$\sub$with/=-0.11), Palmitoleic (bias$\sub$without/=-0.03 and bias$\sub$with/=0.03), Stearic (bias$\sub$without/=0.47 and bias$\sub$with/=0.28), oleic (bias$\sub$without/=0.14 and bias$\sub$with/=-0.04) and linoleic (bias$\sub$without/=0.25 and bias$\sub$with/=-0.20).

• Development and Reproduction
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• v.12 no.3
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• pp.289-296
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• 2008

BPES (Blepharophimosis/Ptosis/Epicanthus inversus Syndrome) is an autosomal dominant disorder caused by mutations in FOXL2. Affected individuals have premature ovarian failure (POF) in addition to small palpebral fissures, drooping eyelids, and broad nasal bridge. FOXL2 is a member of the forkhead family transcription factors. In FOXL2-deficient ovaries, granulosa cell differentiation dose not progress, leading to arrest of folliculogenesis and oocytes atresia. Using yeast two-hybrid screening of rat ovarian cDNA library with FOXL2 as bait, we found that small ubiquitin-related modifier (SUMO)-conjugating E2 enzyme UBE2I protein interacted with FOXL2 protein. UBE2I also known as UBC9 is an essential protein for processing SUMO modification. Sumoylation is a form of post-translational modification involved in diverse signaling pathways including the regulation of transcriptional activities of many transcriptional factors. In the present study, we confirmed the protein-protein interaction between FOXL2 and UBE2I in human cells, 293T, by in vivo immunoprecipitation. In addition, we generated truncated FOXL2 mutants and identified the region of FOXL2 required for its association with UBE2I using yeast-two hybrid system. Therefore, the identification of UBE2I as an interacting protein of FOXL2 further suggests a presence of novel regulatory mechanism of FOXL2 by sumoylation.

• Journal of Applied Biological Chemistry
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• v.65 no.4
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• pp.421-427
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• 2022

In this study, Jeju Tatary buckwheat tea's chemical composition and physiological activities were compared according to the leaching temperature (60, 80, 100 ℃). As the leaching temperature is increased, the degree of browning is induced. However, there was no significant change in pH. The total polyphenol content was higher at 80 ℃ than at 60 ℃ leaching temperature, but significantly decreased at 100 ℃ leaching temperature (60 ℃: 17.06 mg GA/g, 80 ℃: 20.09 mg GA/g, 100 ℃ :18.45 mg GA/g). There were high content of flavonoid and rutin as the leaching temperature increased. Consistently, 2,2-diphenyl1-picrylhydrazyl (DPPH) radical scavenging activity and tyrosinase inhibitory activity were significantly higher with increasing temperature (DPPH % inhibition: 60 ℃: 41.88%, 80 ℃: 46.01%, 100 ℃: 46.80%/tyrosinase inhibitory activity: 60 ℃: 9.38%, 80 ℃: 22.94%, 100 ℃: 28.17%). However, there was no significant difference in DPPH radical scavenging activity between 80 and 100 ℃. A cytotoxicity test was performed by treating with Jeju Tatary buckwheat extract into mouse macrophage cells (Raw264.7). 100 and 200 ㎍/mL treatment (100 ℃ extract) were significantly upregulated the survival rate, but there was no significant difference in other concentrations. Collectively, most of the bioactive components, antioxidant activity, and tyrosinase inhibitory activity were induced as the leaching temperature increased. However, the content of polyphenols which are known to have antioxidant activity, was significantly reduced at 100 ℃ leaching temperature. Several reports have demonstrated that leaching at too high temperature lowered the overall acceptability, so the optimal leaching condition of Tatary Buckwheat is 80 ℃, 5 min in this study.

• Journal of Life Science
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• v.33 no.1
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• pp.1-7
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• 2023

Intracellular cargo transport is mediated by molecular motor proteins, such as kinesin and cytoplasmic dynein. Kinesins make up a large subfamily of molecular motors. Kinesin-1 is a plus-end-directed molecular motor protein that moves various cargoes, such as organelles, protein complexes, and mRNAs, along a microtubule track. It consists of the kinesin superfamily protein (KIF) 5A, 5B, and 5C (also called kinesin heavy chains) and kinesin light chains (KLCs). Kinesin-1 interacts with many different binding proteins through its carboxyl (C)-terminal region of KIF5s and KLCs, but their binding proteins have not yet been fully identified. In this study, a yeast two-hybrid assay was used to identify the proteins that interact with the KIF5A specific C-terminal region. The assay revealed an interaction between KIF5A and glutamate-rich 4 (ERICH4). ERICH4 bound to the KIF5A specific the C-terminal region but did not interact with the C-terminal region of KIF5B or KIF3A (a motor protein of kinesin-2). In addition, KIF5A did not interact with another isoform, ERICH1. Glutathione S-transferase (GST) pull-downs showed that KIF5A interacts with GST-ERICH4 and GST-ERICH4-amino (N)-terminal but not with GST-ERICH4-C or GST alone. When co-expressed in HEK-293T cells, ERICH4 co-localized with KIF5A and co-immunoprecipitated with KIF5A and KLC but not KIF3B. Together, our findings suggest that ERICH4 is capable of binding to KIF5A and that it may serve as an adaptor protein that links kinesin-1 with cargo.

• Molecules and Cells
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• v.45 no.12
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• pp.896-910
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• 2022

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a highly transmissible and potentially fatal virus. So far, most comprehensive analyses encompassing clinical and transcriptional manifestation have concentrated on the lungs. Here, we confirmed evident signs of viral infection in the lungs and spleen of SARS-CoV-2-infected K18-hACE2 mice, which replicate the phenotype and infection symptoms in hospitalized humans. Seven days post viral detection in organs, infected mice showed decreased vital signs, leading to death. Bronchopneumonia due to infiltration of leukocytes in the lungs and reduction in the spleen lymphocyte region were observed. Transcriptome profiling implicated the meticulous regulation of distress and recovery from cytokine-mediated immunity by distinct immune cell types in a time-dependent manner. In lungs, the chemokine-driven response to viral invasion was highly elevated at 2 days post infection (dpi). In late infection, diseased lungs, post the innate immune process, showed recovery signs. The spleen established an even more immediate line of defense than the lungs, and the cytokine expression profile dropped at 7 dpi. At 5 dpi, spleen samples diverged into two distinct groups with different transcriptome profile and pathophysiology. Inhibition of consecutive host cell viral entry and massive immunoglobulin production and proteolysis inhibition seemed that one group endeavored to survive, while the other group struggled with developmental regeneration against consistent viral intrusion through the replication cycle. Our results may contribute to improved understanding of the longitudinal response to viral infection and development of potential therapeutics for hospitalized patients affected by SARS-CoV-2.

• Journal of the Korean Applied Science and Technology
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• v.40 no.3
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• pp.534-546
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• 2023

In this study, we investigated the antioxidant and anti-winkle activity in human fibroblast cell (CCD-986sk) of Persimmon Leaves (PL) as a cosmetic ingredient. As a result of investigating antioxidant activity through electron-donating ability and ABTS⁺ radical scavenging assay, the PL showed concentration-dependent antioxidant activity similar to ascorbic acid, a control group, at a concentration of 1,000 ㎍/ml. As a result of investigating the anti-wrinkle effect through elastase inhibition and collagenase inhibition assay, the PL showed concentration-dependent antioxidant activity similar to epigallocatechin gallate, a control group, at a concentration of 1,000 ㎍/ml. As a result of measuring the synthesis rate of pro-collagen type I and the inhibition rate of MMP-1 in UVB-induced CCD-986sk cells, the control group EGCG showed a 90.2% pro-collagen synthesis rate at 20 ㎍/ml and PL showed an 88.5% synthesis rate at 30 ㎍/ml. In addition, the inhibition rate of MMP-1 of 33.0% and 40.8% were confirmed in EGCG 20 ㎍/ml and PL 30 ㎍/ml, respectively. As a result of measuring the protein expression of pro-collagen type I and MMP-1 in the PL through western blot, it was confirmed that the protein expression of pro-collagen type I increased, and MMP-1 decreased when the PL was treated together compared to the UVB alone group. According to the above experimental results, it is expected to be used as a natural product material for cosmetics by confirming that the PL prevent photoaging caused by UVB stimulation and have antioxidant and anti-wrinkle effects.

• Korean Journal of Ecology and Environment
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• v.56 no.1
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• pp.36-44
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• 2023

Environmental DNA (eDNA) can exist in both intracellular and extracellular forms in natural ecosystems. When targeting harmful cyanobacteria, extracellular eDNA indicates the presence of traces of cyanobacteria, while intracellular eDNA indicates the potential for cyanobacteria to occur. However, identifying the "actual" potential for harmful cyanobacteria to occur is difficult using the existing sediment eDNA analysis method, which uses silica beads and cannot distinguish between these two forms of eDNA. This study analyzes the applicability of a density gradient centrifugation method (Ludox method) that can selectively analyze intracellular eDNA in sediment to overcome the limitations of conventional sediment eDNA analysis. PCR was used to amplify the extracted eDNA based on the two different methods, and the relative amount of gene amplification was compared using electrophoresis and Image J application. While the conventional bead beating method uses sediment as it is to extract eDNA, it is unknown whether the mic gene amplified from eDNA exists in the cyanobacterial cell or only outside of the cell. However, since the Ludox method concentrates the intracellular eDNA of the sediment through filtration and density gradient, only the mic gene present in the cyanobacteria cells could be amplified. Furthermore, the bead beating method can analyze up to 1 g of sediment at a time, whereas the Ludox method can analyze 5 g to 30 g at a time. This gram of sediments makes it possible to search for even a small amount of mic gene that cannot be searched by conventional bead beating method. In this study, the Ludox method secured sufficient intracellular gene concentration and clearly distinguished intracellular and extracellular eDNA, enabling more accurate and detailed potential analysis. By using the Ludox method for environmental RNA expression and next-generation sequencing (NGS) of harmful cyanobacteria in the sediment, it will be possible to analyze the potential more realistically.

• Journal of Life Science
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• v.33 no.2
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• pp.158-168
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• 2023

Natural products have been used to mitigate the effects of cancer and infectious diseases, as they feature diverse bioactivities, such as antioxidant, antibacterial, anti-inflammatory, and immunomodulatory effects. Here, we chose 10 natural products that are well-known as pulmonary enhancers and investigated their bactericidal effects on Streptococcus pneumoniae. In the disk diffusion assay, the growth of S. pneumoniae was significantly regulated by G. fructus treatment regardless of extraction method used. We first adopted spraying as a novel delivery method for G. fructus. Interestingly, mice exposed to G. fructus three times a day for 2 weeks were resistant to S. pneumoniae intranasal infection (shown both through body weight loss and survival rates compared to the control group). Moreover, we confirmed that exposure to G. fructus regulated the colonization of the bacteria despite the sustained inflammation in the lung after exposure to S. pneumoniae, indicating that migrated inflammatory immune cells may involve a host defense mechanism against pulmonary infectious diseases. While a similar number of granulocytes (CD11b⁺Ly6C⁺Ly6G⁺), neutrophils (CD11b⁺Ly6C^intLy6G⁺), and monocytes (CD11b⁺Ly6C^intLy6G^-) were found between groups, a significantly increased number of alveolar macrophages (CD11b⁺CD11c^hiF4/80⁺) was detected in BAL fluids of mice pre-exposed to G. fructus at 5 days after S. pneumonia infection. Taken together, our data suggest that this usage of G. fructus can induce protective immunity against bacterial infection, indicating that facial spray may be helpful in enhancing the defense mechanism against pulmonary inflammation and in evaluating the efficacy of natural products as immune enhancers against respiratory diseases.