• Journal of Ginseng Research
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• v.43 no.1
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• pp.116-124
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• 2019

Background: Ginsenosides are known as the principal pharmacological active constituents in Panax medicinal plants such as Asian ginseng, American ginseng, and Notoginseng. Some ginsenosides, especially the 20(R) isomers, are found in trace amounts in natural sources and are difficult to chemically synthesize. The present study provides an approach to produce such trace ginsenosides applying biotransformation through Escherichia coli modified with relevant genes. Methods: Seven uridine diphosphate glycosyltransferase (UGT) genes originating from Panax notoginseng, Medicago sativa, and Bacillus subtilis were synthesized or cloned and constructed into pETM6, an ePathBrick vector, which were then introduced into E. coli BL21star (DE3) separately. 20(R)-Protopanaxadiol (PPD), 20(R)-protopanaxatriol (PPT), and 20(R)-type ginsenosides were used as substrates for biotransformation with recombinant E. coli modified with those UGT genes. Results: E. coli engineered with $GT95^{syn}$ selectively transfers a glucose moiety to the C20 hydroxyl of 20(R)-PPD and 20(R)-PPT to produce 20(R)-CK and 20(R)-F1, respectively. GTK1- and GTC1-modified E. coli glycosylated the C3-OH of 20(R)-PPD to form 20(R)-Rh2. Moreover, E. coli containing $p2GT95^{syn}K1$, a recreated two-step glycosylation pathway via the ePathBrich, implemented the successive glycosylation at C20-OH and C3-OH of 20(R)-PPD and yielded 20(R)-F2 in the biotransformation broth. Conclusion: This study demonstrates that rare 20(R)-ginsenosides can be produced through E. coli engineered with UTG genes.

• Animal Bioscience
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• v.34 no.11
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• pp.1739-1748
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• 2021

Objective: In recent years, long noncoding RNAs (lncRNAs) have been identified in many species, and some of them have been shown to play important roles in muscle development and myogenesis. However, the differences in lncRNAs between Kazakh cattle and Xinjiang brown cattle remain undefined; therefore, we aimed to confirm whether lncRNAs are differentially expressed in the longissimus dorsi between these two types of cattle and whether differentially expressed lncRNAs regulate muscle differentiation. Methods: We used RNA-seq technology to identify lncRNAs in longissimus muscles from these cattle. The expression of lncRNAs were analyzed using StringTie (1.3.1) in terms of the fragments per kilobase of transcript per million mapped reads values of the encoding genes. The differential expression of the transcripts in the two samples were analyzed using the DESeq R software package. The resulting false discovery rate was controlled by the Benjamini and Hochberg's approach. KOBAS software was utilized to measure the expression of different genes in Kyoto encyclopedia of genes and genomes pathways. We randomly selected eight lncRNA genes and validated them by quantitative reverse transcription polymerase chain reaction (RT-qPCR). Results: We found that 182 lncRNA transcripts, including 102 upregulated and 80 downregulated transcripts, were differentially expressed between Kazakh cattle and Xinjiang brown cattle. The results of RT-qPCR were consistent with the sequencing results. Enrichment analysis and functional annotation of the target genes revealed that the differentially expressed lncRNAs were associated with the mitogen-activated protein kinase, Ras, and phosphatidylinositol 3-kinase (PI3k)/Akt signaling pathways. We also constructed a lncRNA/mRNA coexpression network for the PI3k/Akt signaling pathway. Conclusion: Our study provides insights into cattle muscle-associated lncRNAs and will contribute to a more thorough understanding of the molecular mechanism underlying muscle growth and development in cattle.

• Biomolecules & Therapeutics
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• v.30 no.3
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• pp.274-283
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• 2022

KRAS activating mutations, which are present in more than 90% of pancreatic cancers, drive tumor dependency on the RAS/mitogen-activated protein kinase (MAPK) and phosphoinositide 3-kinase (PI3K)/AKT signaling pathways. Therefore, combined targeting of RAS/MAPK and PI3K/AKT signaling pathways may be required for optimal therapeutic effect in pancreatic cancer. However, the therapeutic efficacy of combined MAPK and PI3K/AKT signaling target inhibitors is unsatisfactory in pancreatic cancer treatment, because it is often accompanied by MAPK pathway reactivation by PI3K/AKT inhibitor. Therefore, we developed an inRas37 antibody, which directly targets the intra-cellularly activated GTP-bound form of oncogenic RAS mutation and investigated its synergistic effect in the presence of the PI3K inhibitor BEZ-235 in pancreatic cancer. In this study, inRas37 remarkably increased the drug response of BEZ-235 to pancreatic cancer cells by inhibiting MAPK reactivation. Moreover, the co-treatment synergistically inhibited cell proliferation, migration, and invasion and exhibited synergistic anticancer activity by inhibiting the MAPK and PI3K pathways. The combined administration of inRas37and BEZ-235 significantly inhibited tumor growth in mouse models. Our results demonstrated that inRas37 synergistically increased the antitumor activity of BEZ-235 by inhibiting MAPK reactivation, suggesting that inRas37 and BEZ-235 co-treatment could be a potential treatment approach for pancreatic cancer patients with KRAS mutations.

• Korean journal of applied entomology
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• v.61 no.1
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• pp.173-195
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• 2022

Like vertebrates, insects synthesize various eicosanoids after the committed catalytic step of phospholipase A₂ (PLA₂). However, the subsequent biosynthetic steps exhibit some deviation from those of vertebrates. Due to little composition of arachidonic acid in insect phospholipids, PLA₂ releases linoleic acid, which is another polyunsaturated fatty acid and relatively rich in insect phospholipids, to synthesize arachidonic acid via chain extension and desaturation. Resulting arachidonic acid is then oxygenated into a prostaglandin (PG), PGH₂, by a specific peroxidase called peroxynectin, but not by cyclooxygenase. PGH₂ is then isomerized to various PGs such as PGA₂, PGD₂, PGE₂, PGI₂, and a thromboxane (TXB₂). All four epoxyeicosatrienoic acids such as 5,6-EET, 8,9-EET, 11,12-EET, and 14,15-EET are also synthesized from arachidonic acid by oxygenation of vertebrate types of monooxygenases. However, the other type of eicosanoids called leukotrienes are found in insect tissues but their synthetic pathway is unclear. Eicosanoids mediate various insect physiological processes such as metabolism, excretion, immunity, and reproduction. Thus, identification of novel compounds interrupting eicosanoid biosynthesis would be a novel approach to develop insecticides. This review focuses on PGs and their immune mediation.

• Journal of Veterinary Science
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• v.24 no.4
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• pp.52.1-52.13
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• 2023

Background: Mesenchymal stem cells (MSCs) have been investigated as therapeutic agents for inflammatory bowel disease (IBD). Stimulation of MSCs with pro-inflammatory cytokines is an approach to enhance their immunomodulatory effects. However, further investigation is required to support their application in immune-mediated disorders and companion animals. Objectives: This study aimed to assess the therapeutic effect of tumor necrosis factor (TNF)-α-stimulated feline adipose tissue-derived MSCs (fAT-MSCs) in a dextran sulfate sodium (DSS)-induced colitis mouse model. Methods: Colitis mice was made by drinking water with 3% DSS and fAT-MSCs were injected intraperitoneally. Colons were collected on day 10. The severity of the disease was evaluated and compared. Raw 264.7 cells were cultured with the conditioned medium to determine the mechanism, using quantitative real-time polymerase chain reaction and enzyme-linked immunosorbent assay. Results: TNF-α-stimulated fAT-MSCs more improved severity of DSS-induced colitis in disease activity, colon length, histologic score, and inflammatory cytokine. In sectionized colon tissues, the group comprising TNF-α-stimulated fAT-MSCs had higher proportion of CD11b⁺CD206⁺ macrophages than in the other groups. In vitro, TNF-α-stimulation increased cyclooxygenase-2 (COX-2) expression and prostaglandin E₂ (PGE₂) secretion from fAT-MSCs. The conditioned medium from TNF-α-stimulated fAT-MSCs enhanced the expression of interleukin-10 and arginase-1 in LPS-activated Raw 264.7 cells. Conclusions: These results represent that TNF-α-stimulated fat-mscs ameliorate the inflamed colon more effectively. Furthermore, we demonstrated that the effectiveness was interlinked with the COX-2/PGE₂ pathway.

• Korean Journal of Agricultural and Forest Meteorology
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• v.25 no.4
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• pp.284-301
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• 2023

The changes in rice climatic yield potential (CYP) across the Korean Peninsula are evaluated based on the new climate change scenario produced by the National Institute of Agricultural Sciences with 18 ensemble members at 1 km resolution under a Shared Socioeconomic Pathway (SSP) and Representative Concentration Pathways (RCP) emission scenarios. To overcome the data availability, we utilize solar radiation f or CYP instead of sunshine duration which is relatively uncommon in the climate prediction f ield. The result show that maximum CYP(CYPmax) decreased, and the optimal heading date is progressively delayed under warmer temperature conditions compared to the current climate. This trend is particularly pronounced in the SSP5-85 scenario, indicating faster warming, except for the northeastern mountainous regions of North Korea. This shows the benef its of lower emission scenarios and pursuing more efforts to limit greenhouse gas emissions. On the other hand, the CYPmax shows a wide range of feasible futures, which shows inherent uncertainties in f uture climate projections and the risks when analyzing a single model or a small number of model results, highlighting the importance of the ensemble approach. The f indings of this study on changes in rice productivity and uncertainties in temperature and solar radiation during the 21st century, based on climate change scenarios, hold value as f undamental information for climate change adaptation efforts.

• Biomolecules & Therapeutics
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• v.32 no.1
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• pp.84-93
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• 2024

Rosmarinic acid (RA) is a phenolic ester that protects human keratinocytes against oxidative damage induced by ultraviolet B (UVB) exposure, however, the mechanisms underlying its effects remain unclear. This study aimed to elucidate the cell signaling mechanisms that regulate the antioxidant activity of RA and confirm its cyto-protective role. To explore the signaling mechanisms, we used the human keratinocyte cell line HaCaT and SKH1 hairless mouse skin. RA enhanced glutamate-cysteine ligase catalytic subunit (GCLC) and glutathione synthetase (GSS) expression in HaCaT cells in a dose- and time-dependent manner. Moreover, RA induced nuclear factor erythroid-2-related factor 2 (NRF2) nuclear translocation and activated the signaling kinases protein kinase B (AKT) and extracellular signal-regulated kinase (ERK). Treatment with the phosphatidylinositol 3-kinase (PI3K) inhibitor LY294002, the ERK inhibitor U0126, and small interfering RNA (siRNA) gene silencing suppressed RA-enhanced GCLC, GSS, and NRF2 expression, respectively. Cell viability tests showed that RA significantly prevented UVB-induced cell viability decrease, whereas the glutathione (GSH) inhibitors buthionine sulfoximine, LY294002, and U0126 significantly reduced this effect. Moreover, RA protected against DNA damage and protein carbonylation, lipid peroxidation, and apoptosis caused by UVB-induced oxidative stress in a concentration-dependent manner in SKH1 hairless mouse skin tissues. These results suggest that RA protects against UVB-induced oxidative damage by activating AKT and ERK signaling to regulate NRF2 signaling and enhance GSH biosynthesis. Thus, RA treatment may be a promising approach to protect the skin from UVB-induced oxidative damage.

• Journal of Life Science
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• v.25 no.6
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• pp.693-702
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• 2015

Blocking new blood-vessel formation (angiogenesis) is now recognized as a useful approach to the therapeutic treatment of many solid tumors. The best validated approach to date is to target the vascular endothelial growth-factor (VEGF) pathway, a key regulator of angiogenesis. Many natural products and extracts that contain a variety of chemopreventive compounds have been shown to suppress the development of malignancies through their anti-angiogenic properties. Phellodendron amurense, which is widely used in Korean traditional medicine, has been shown to possess antitumor, antimicrobial, and anti-inflammatory properties, among others. The present study investigated the effects of P. amurense hot-water extract (PAHWE) on angiogenesis, a key process in tumor growth, invasion, and metastasis. To investigate PAHWE’s anti-angiogenic properties, this study’s authors performed an analysis of angiogenesis and endothelial-cell proliferation, migration, invasion, and tube formation, as well as zymogram assays and the rat aortic ring-sprouting assay. PAHWE inhibited cell growth, mobility, and vessel formation in response to VEGF in vitro and ex vivo. Furthermore, it reduced VEGF-induced intracellular signaling events, such as the activation of matrix metalloproteinases (MMPs) -2 and -9. These results indicate that PAHWE’s anti-angiogenic properties might lead to the development of potential drugs for treating angiogenesis-associated diseases such as cancer.

• Korean Journal of Food Science and Technology
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• v.37 no.3
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• pp.484-491
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• 2005

Quantitative microbial risk assessment (QMRA) analyzes potential hazard of microorganisms on public health and offers structured approach to assess risks associated with microorganisms in foods. This paper addresses specific risk management questions associated with Staphylococcus aureus in kimbab and improvement and dissemination of QMRA methodology, QMRA model was developed by constructing four nodes from retail to table pathway. Predictive microbial growth model and survey data were combined with probabilistic modeling to simulate levels of S. aureus in kimbab at time of consumption, Due to lack of dose-response models, final level of S. aureus in kimbeb was used as proxy for potential hazard level, based on which possibility of contamination over this level and consumption level of S. aureus through kimbab were estimated as 30.7% and 3.67 log cfu/g, respectively. Regression sensitivity results showed time-temperature during storage at selling was the most significant factor. These results suggested temperature control under $10^{\circ}C$ was critical control point for kimbab production to prevent growth of S. aureus and showed QMRA was useful for evaluation of factors influencing potential risk and could be applied directly to risk management.

• Journal of Environmental Policy
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• v.13 no.1
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• pp.69-93
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• 2014

The aim of this research was to develop a climate change vulnerability index at the district level (Si, Gun, Gu) with respect to the health care sector in Korea. The climate change vulnerability index was esimated based on the four major causes of climate-related illnesses : vector, flood, heat waves, and air pollution/allergies. The vulnerability assessment framework consists of six layers, all of which are based on the IPCC vulnerability concepts (exposure, sensitivity, and adaptive capacity) and the pathway of direct and indirect impacts of climate change modulators on health. We collected proxy variables based on the conceptual framework of climate change vulnerability. Data were standardized using the min-max normalization method. We applied the analytic hierarchy process (AHP) weight and aggregated the variables using the non-compensatory multi-criteria approach. To verify the index, sensitivity analysis was conducted by using another aggregation method (geometric transformation method, which was applied to the index of multiple deprivation in the UK) and weight, calculated by the Budget Allocation method. The results showed that it would be possible to identify the vulnerable areas by applying the developed climate change vulnerability assessment index. The climate change vulnerability index could then be used as a valuable tool in setting climate change adaptation policies in the health care sector.