• Biomolecules & Therapeutics
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• v.31 no.6
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• pp.682-691
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• 2023

Cell transformation induced by epidermal growth factor (EGF) and 12-O-tetradecanoylphorbol-13-acetate (TPA) is a critical event in cancer initiation and progression, and understanding the underlying mechanisms is essential for the development of new therapeutic strategies. Licorice extract contains various bioactive compounds, which have been reported to have anticancer and anti-inflammatory effects. This study investigated the cancer preventive efficacy of licochalcone D (LicoD), a chalcone derivative in licorice extract, in EGF and TPA-induced transformed skin keratinocyte cells. LicoD effectively suppressed EGF-induced cell proliferation and anchorage-independent colony growth. EGF and TPA promoted the S phase of cell cycle, while LicoD treatment caused G1 phase arrest and down-regulated cyclin D1 and up-regulated p21 expression associated with the G1 phase. LicoD also induced apoptosis and increased apoptosis-related proteins such as cleaved-caspase-3, cleaved-caspase-7, and Bax (Bcl2-associated X protein). We further investigated the effect of LicoD on the AKT signaling pathway involved in various cellular processes and found decreased p-AKT, p-GSK3β, and p-NFκB expression. Treatment with MK-2206, an AKT pharmacological inhibitor, suppressed EGF-induced cell proliferation and transformed colony growth. In conclusion, this study demonstrated the potential of LicoD as a preventive agent for skin carcinogenesis.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2022.09a
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• pp.90-90
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• 2022

Pueraria flos and Adzuki Beans contains several bioactive compounds, such as saponin, oleanolic acid, and flavone. Pueraria flos and Adzuki Beans has traditionally been used to treat disorders of antioxidant activity, diabetes and liver detoxication, and it has antinociceptive and anti-inflammatory properties. However, complex saponin were validity of the anti-inflammatory activity has not been scientifically investigated. In this study, to determine anti-inflammatory activity of complex saponin separated from Pueraria flos and Adzuki Beans mixture extract on nitric oxide and prostaglandinE₂ assay. The anti-inflammatory activities of complex saponin separated from Pueraria flos and Adzuki Beans mixture extract were evaluated for inhibitory activities against lipopolysacchride induced nitric oxide and prostaglandinE₂ production protein expressions in RAW264.7 cell lines. The complex saponin separated from Pueraria flos and Adzuki Beans mixture extract inhibitory activity for both tests with protein high depressions(%) values showed in the ranges of 50~100 ㎍/ml. Overall, prostaglandinE₂ tests had a higher inhibitory effect on inflammation than nitricoxide tests. Theseis result suggest a potential role of complex saponin separated from Pueraria flos and Adzuki Beans mixture extract as source of anti-inflammation agent.

• Animal Bioscience
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• v.37 no.5
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• pp.908-917
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• 2024

Objective: Although pork loins is not a tough meat, they need to develop meat products with a soft texture for the elderly. This study focused on the physicochemical properties and tenderness characteristics of pork loin injected with green kiwifruit juice (GRJ) and gold kiwifruit juice (GOJ) during various incubation times. In addition, the antioxidant activities of hydrolysate derived from the hydrolysis of pork loin by kiwifruit juice protease were evaluated. Methods: The pork loin was injected with 10% and 20% GRJ and GOJ, under various incubation times (0, 4, 8, and 24 h). Then, the physicochemical properties and tenderness of pork loins were measured. 2,2- diphenyl-1-picrylhydrazyl radical scavenging activity and reducing power were conducted to determine hydrolysate's antioxidant activities derived from pork loin's hydrolysis by kiwifruit juice protease. Results: GRJ had greater tenderizing ability than GOJ, even at the 10% addition. When kiwifruit juice was injected into pork loin, the tenderness increased with increasing incubation time. This was confirmed by the decrease in intensity of the myosin heavy chain (MHC) band in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. In particular, the MHC band decreased at 8 h for both 10% GRJ and 20% GOJ and at 4 h for 20% GRJ alone. The highest myofibril fragmentation index and peptide solubility were observed in pork loin treated with 20% GRJ compared to the other treatments during incubation. The 10% GRJ and 20% GOJ treatments showed similar levels of antioxidant activity of the protein hydrolysates in pork loin, and 20% GRJ showed the highest activity among the treatments. Conclusion: Kiwifruit juice had protease activity, and GRJ was more useful for tenderizing meat products than GOJ. Thus, GRJ at 10% could be a potential agent to tenderize and enrich the natural antioxidant activity through the proteolysis of pork loin.

• Korean Journal of Acupuncture
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• v.41 no.1
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• pp.7-15
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• 2024

Objectives : This study aimed to predict the effectiveness and potential of Schizonepeta tenuifolia as an anticancer treatment for non-small cell lung cancer through network-based pharmacology and cellular experiment. Methods : To identify the major bioactive compounds in Schizonepeta tenuifolia, we used the Traditional Chinese Medicine Systems. The target genes for the cancer treatment were selected using the UniProt database and the networked using Cytoscape. We performed functional enrichment analysis based on the Gene Ontology Biological Process and Kyoto Encyclopedia of Genes and Genomes Pathways to predict the mechanisms. To investigate the effect of Schizonepeta tenuifolia on lung cancer cell growth, we treated A549 cells, a lung cancer cell line, with different concentrations of the drug and used the MTT assay for cell viability. Results : Research has shown that the most effective mechanism of active compounds from Schizonepeta tenuifolia is through the pathway of cancer. The results of the network pharmacology analysis indicate that Schizonepeta tenuifolia has potential medicinal value as an adjuvant in anticancer treatment. The concentration-dependent inhibition of cell viability was observed on A549 cells. Furthermore, synergistic anticancer activity with Doxorubicin was also observed. Conclusions : Through a network pharmacological approach, Schizonepeta tenuifolia was predicted to have potential as an anticancer agent, and its efficacy was experimentally demonstrated using A549 cells. These findings suggest that Schizonepeta tenuifolia is a promising candidate for future research.

• Journal of Veterinary Clinics
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• v.31 no.6
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• pp.502-506
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• 2014

Recently nano particles have proven for wide array of bioactive properties. In the present study, antibacterial properties of chitosan silver nano composites (CAgNCs) were investigated against fish pathogenic Edwardsiella tarda. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of CAgNCs against E. tarda were $25{\mu}g/mL$ and $125{\mu}g/mL$, respectively. The field emission scanning electron microscope (FE-SEM) image of CAgNCs treated E. tarda showed the strongly damaged bacteria cells than non-treated bacteria. Furthermore, treatment of CAgNCs induced the level of intracellular reactive oxygen species (ROS) in E. tarda cells in concentration and time dependent manner suggesting that it may generate oxidative stress leading to bacterial cell death. In addition, MTT assay results showed that the lowest cell viability at $100{\mu}g/mL$ of CAgNCs treated E. tarda. Overall results of this study suggest that CAgNCs is a potential antibacterial agent to control pathogenic bacteria.

• 한국균학회소식:학술대회논문집
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• 2014.05a
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• pp.27-28
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• 2014

Beauveria bassiana (Cordycipitaceae, Hypocreales, Ascomycota) is an anamorphic fungus having a potential to be used as a biological control agent because it parasitizes a wide range of arthropod hosts including termites, aphids, beetles and many other insects. A number of bioactive secondary metabolites (SMs) have been isolated from B. bassiana and functionally verified. Among them, beauvericin and bassianolide are cyclic depsipeptides with antibiotic and insecticidal effects belonging to the enniatin family. Non-ribosomal peptide synthetases (NRPSs) play a crucial role in the synthesis of these secondary metabolites. NRPSs are modularly organized multienzyme complexes in which each module is responsible for the elongation of proteinogenic and non-protein amino acids, as well as carboxyl and hydroxyacids. A minimum of three domains are necessary for one NRPS elongation module: an adenylation (A) domain for substrate recognition and activation; a tholation (T) domain that tethers the growing peptide chain and the incoming aminoacyl unit; and a condensation (C) domain to catalyze peptide bond formation. Some of the optional domains include epimerization (E), heterocyclization (Cy) and oxidation (Ox) domains, which may modify the enzyme-bound precursors or intermediates. In the present study, we analyzed genomes of B. bassiana and its allied species in Hypocreales to verify the distribution of NRPS-encoding genes involving biosynthesis of beauvericin and bassianolide, and to unveil the evolutionary processes of the gene clusters. Initially, we retrieved completely or partially assembled genomic sequences of fungal species belonging to Hypocreales from public databases. SM biosynthesizing genes were predicted from the selected genomes using antiSMASH program. Adenylation (A) domains were extracted from the predicted NRPS, NRPS-like and NRPS-PKS hybrid genes, and used them to construct a phylogenetic tree. Based on the preliminary results of SM biosynthetic gene prediction in B. bassiana, we analyzed the conserved gene orders of beauvericin and bassianolide biosynthetic gene clusters among the hypocrealean fungi. Reciprocal best blast hit (RBH) approach was performed to identify the regions orthologous to the biosynthetic gene cluster in the selected fungal genomes. A clear recombination pattern was recognized in the inferred A-domain tree in which A-domains in the 1st and 2nd modules of beauvericin and bassianolide synthetases were grouped in CYCLO and EAS clades, respectively, suggesting that two modules of each synthetase have evolved independently. In addition, inferred topologies were congruent with the species phylogeny of Cordycipitaceae, indicating that the gene fusion event have occurred before the species divergence. Beauvericin and bassianolide synthetases turned out to possess identical domain organization as C-A-T-C-A-NM-T-T-C. We also predicted precursors of beauvericin and bassianolide synthetases based on the extracted signature residues in A-domain core motifs. The result showed that the A-domains in the 1st module of both synthetases select D-2-hydroxyisovalerate (D-Hiv), while A-domains in the 2nd modules specifically activate L-phenylalanine (Phe) in beauvericin synthetase and leucine (Leu) in bassianolide synthetase. antiSMASH ver. 2.0 predicted 15 genes in the beauvericin biosynthetic gene cluster of the B. bassiana genome dispersed across a total length of approximately 50kb. The beauvericin biosynthetic gene cluster contains beauvericin synthetase as well as kivr gene encoding NADPH-dependent ketoisovalerate reductase which is necessary to convert 2-ketoisovalarate to D-Hiv and a gene encoding a putative Gal4-like transcriptional regulator. Our syntenic comparison showed that species in Cordycipitaceae have almost conserved beauvericin biosynthetic gene cluster although the gene order and direction were sometimes variable. It is intriguing that there is no region orthologous to beauvericin synthetase gene in Cordyceps militaris genome. It is likely that beauvericin synthetase was present in common ancestor of Cordycipitaceae but selective gene loss has occurred in several species including C. militaris. Putative bassianolide biosynthetic gene cluster consisted of 16 genes including bassianolide synthetase, cytochrome P450 monooxygenase, and putative Gal4-like transcriptional regulator genes. Our synteny analysis found that only B. bassiana possessed a bassianolide synthetase gene among the studied fungi. This result is consistent with the groupings in A-domain tree in which bassianolide synthetase gene found in B. bassiana was not grouped with NRPS genes predicted in other species. We hypothesized that bassianolide biosynthesizing cluster genes in B. bassiana are possibly acquired by horizontal gene transfer (HGT) from distantly related fungi. The present study showed that B. bassiana is the only species capable of producing both beauvericin and bassianolide. This property led to B. bassiana infect multiple hosts and to be a potential biological control agent against agricultural pests.

• Journal of Microbiology and Biotechnology
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• v.27 no.12
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• pp.2156-2164
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• 2017

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is considered as an antitumor agent owing to its ability to induce apoptosis of cancer cells without imparting toxicity toward most normal cells. TRAIL is produced in poor yield because of its insoluble expression in the cytoplasm of E. coli. In this study, we achieved soluble expression of TRAIL by fusing maltose-binding protein (MBP), b'a' domain of protein disulfide isomerase (PDIb'a'), or protein disulfide isomerase at the N-terminus of TRAIL. The TRAIL was purified using subsequent immobilized metal affinity chromatography and amylose-binding chromatography, with the tag removal using tobacco etch virus protease. Approximately 4.5 mg of pure TRAIL was produced from 125 ml flask culture with a purification yield of 71.6%. The endotoxin level of the final product was $0.4EU/{\mu}g$, as measured by the Limulus amebocyte lysate endotoxin assay. The purified TRAIL was validated and shown to cause apoptosis of HeLa cells with an $EC_{50}$ and Hill coefficient of $0.6{{\pm}}0.03nM$ and $2.41{\pm}0.15$, respectively. The high level of apoptosis in HeLa cells following administration of purified TRAIL indicates the significance and novelty of this method for producing high-grade and high-yield TRAIL.

• Korean Journal of Head & Neck Oncology
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• v.17 no.2
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• pp.155-161
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• 2001

Background and Objectives: Nitric oxide (NO) is generated in mammalian tissue by the conversion of L-arginine to L-citrulline. This reaction is catalyzed by nitric oxide synthase (NOS). NO is an important bioactive agent and a signalling molecule that mediates a variety of biologic actions such as vasodilation, neurotransmission, host defense, and iron metabolism but increased NO production may also contribute to the pathogenesis of a various of disorders, including cancer. Before now, the role of NO in thyroid gland is still investigated and it was supposed that NO mediate the angiogenesis in tumor growth. Others journal and works identified the expression of iNOS that involve by neutrophil and eNOS that involve in part in the vascular remodeling and to understand the role of NO in human thyroid gland. But authors revealed only eNOS in thyroid neoplasm. iNOS was identifed by inflammation in fault. Materials and Methods: Western blot analysis was performed, using a polyclonal antibody against eNOS (Rabbit polyclonal IgG). Using the same antibody, the distribution of eNOS was examined in 15 formalin-fixed paraffin embedded samples by immunohistochemistry. By NADPH consumption rate, NOS activity was estimated at nodular hyperplasia. Results: Western blot analysis exhibited that eNOS was significantly elevated in thyroid papillary carcinoma, compared to that in nodular hyperplasia and normal tissue. Immunohistochemistry showed that the immunoreacitivity was present more significantly in thyroid follicular epithelial cell layer than vascular endothelial cell. NOS activity increased in nodular hyperplasia. Conclusions: Thyroid papillary cancer without neutrophil invasion expressed only eNOS. The endothelial localization of eNOS may play an important role in pathogenensis of human thyroid nodular hyperplasia and the follicular localization of thyroid papillary carcinomas.

• Journal of Conservation Science
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• v.23
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• pp.95-102
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• 2008

To investigate bioactive materials for development of natural conservative agent on organic cultural heritage, methanol extracts from 11 wood species which populate in Korea were screened for antifungal and insecticidal activity against 9 wood contaminant fungi and adult of Lasioderma serricorne. Antifungal activity of extracts was tested by using paper disc soaking method and feeder strip test against wood-rot fungi and wood surface contaminant fungi. Among these extracts, the most significant antimicrobial activity was observed from the extract of Phellodendron amurense at 5 mg/disc against Coniophora puteana, Lentinus lepideus, Tyromyces palustris and Aspergillus niger. The insecticidal activity of extracts was examined by topical application against L. serricorne adults. The extracts from Taxus cuspidata and Abies holophylla gave 56.7 % mortality at 25 ${\mu}g$/adult for 24 hr. From these result, the methanol extract from T. cuspidata, A. holophylla and P. amurense described could be useful for conservation of organic cultural heritage against biological deterioration by insect and wood contaminant fungi.

• Korean Journal of Veterinary Service
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• v.32 no.4
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• pp.353-359
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• 2009

Interest in marine organisms as potential sources of bioactive agents has increased in recent years. The red seaweed, Callophyllis (C.) japonica, is abundant in the coastal regions of Jeju Island in South Korea. A previous study shows that C. japonica extracts have antioxidant activity and radioprotective effects. In this study, an methanol extract of C. japonica was tested whether it has antibacterial effects against the bacteria from swine. In vitro antibacterial activities of the crude extracts prepared from the C. japonica using 80 % methanol were tested for inhibitory activity against the Escherichia (E.) coli (S175), Enterococcus (E.) faecium (ATCC 51558), Salmonella (S.) Typhimurium and Staphylo-coccus (S.) aureus (ATCC 25923) by using broth dilution method. All organisms were incubated in brain heart infusion medium containing 1% extract at 0, 4, 8, 12 and 24 hrs. The 3 days-old piglets were fed an experimental diet supplemented with 1% C. japonica for 1 week. And the change of the coliform bacteria in feces were examined after supplement of C. japonica for 1 week. When the inocula containing $10^2{\sim}10^3$CFU/ml of each organism were used the extracts of C. japonica showed various degrees of antibacterial effects on all bacteria tested. The CFU value ($6.3\times10^8$CFU/ml) of C. japonica for E. coli was decreased 30% compared with vehicle controls ($9.0\times10^8$CFU/ml) after 8 hrs incubation. The proliferation rate of E. faecium was inhibited about 68% at 4 hrs, 81% at 8 hrs and 76% at 12 hrs after incubation, respectively. The proliferation rate of S. Typhimurium was inhibited about 96% at 4 hrs, 90% at 8 hrs and 72% at 12 hrs after incubation with extracts of C. japonica. The proliferation rate of S. aureus was inhibited more than 90% each time courses. Conclusively, a red seaweed extract of C. japonica was found to be effective against a number of gram negative and gram positive bacteria such as E. coli, E. faecium, S. Typhimurium, and S. aureus. The number of coliform bacteria was increased in the 1% C. japonica-treated group, as compared to those of controls. This result suggests that C. japonica extracts be added as an effective natural antibacterial agent. The precise mechanism of antibacterial effects and its application on swine industry remains to be further studied.