• International Journal of Industrial Entomology and Biomaterials
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• 제8권2호
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• pp.161-167
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• 2004

Silkworms sex determination drew high attention from researchers. Sex chromosomes on the silkworm are of ZW type for females and ZZ type for males. Chromosome W plays an important role in sex determination. Although several molecular linkage maps have been constructed for silkworm, very few markers are discovered on the W chromosome. In order to look for molecular markers and to further locate the Fern gene on chromosome W, we used genomic DNA from both female and male larvae of a silkworm strain named 937 as PCR templates for RAPD amplification with 200 arbitrary 10-mer primers. The amplification results showed three female-specific bands, namely ${OPG-07_496}, {OPC-15_1,660} and {OPE-18_1,279}$. Further verification, however, revealed no band from OPG-07 and OPC-15 in either sex in the strain 798, but OPE-18 provided female-specific band in the strains Suluan7 and C108, and absent in both males and strain 798. This indicates that the bands from ${OPG-07_496} and {OPC-15_1,660}$ are probably female-specific in strain 937, and the band from OPE-18 was probably amplified from a common segment shared by most strains. The genomic DNAs from OPG-07 and OPC-15 were cloned and sequenced. Sequence analysis showed that the DNAs from OPG-07 and OPC-15 have high identities with the retrotransposable elements, and DNA from OPC-15 contains a portion of sequence which probably encodes an eukaryotic translation initiation factor 4E binding protein (eIF4EBP).

• The Korean Journal of Physiology and Pharmacology
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• 제13권2호
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• pp.131-138
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• 2009

The binding of interleukin-6 (IL-6) cytokine family ligands to the gp130 receptor complex activates the Janus kinase (JAK)/ signal transducer and activator of transcription 3 (STAT3) signal transduction pathway, where STA T3 plays an important role in cell survival and tumorigenesis. Constitutive activation of STAT3 has been frequently observed in many cancer tissues, and thus, blocking of the gp130 signaling pathway, at the JAK level, might be a useful therapeutic approach for the suppression of STAT3 activity, as anticancer therapy. AG490 is a tyrphostin tyrosine kinase inhibitor that has been extensively used for inhibiting JAK2 in vitro and in vivo. In this study, we demonstrate a novel mechanism associated with AG490 that inhibits the JAK/STAT3 pathway. AG490 induced downregulation of gp130, a common receptor for the IL-6 cytokine family compounds, but not JAK2 or STAT3, within three hours of exposure. The downregulation of gp130 was not caused by enhanced degradation of gp130 or by inhibition of mRNA transcription. It most likely occurred by translation inhibition of gp130 in association with phosphorylation of the eukaryotic initiation factor-2 a. The inhibition of protein synthesis of gp130 by AG490 led to immediate loss of mature gp130 in cell membranes, due to its short half-life, thereby resulting in reduction in the STAT3 response to IL-6. Taken together, these results suggest that AG490 blocks the STAT3 activation pathway via a novel pathway.

• The Korean Journal of Physiology and Pharmacology
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• 제27권4호
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• pp.345-356
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• 2023

This study aimed to assess the effects of exogenous hydrogen sulfide (H₂S) on abdominal aorta coarctation (AAC) induced myocardial fibrosis (MF) and autophagy in rats. Forty-four Sprague-Dawley rats were randomly divided into control group, AAC group, AAC + H₂S group, and H₂S control group. After a model of rats with AAC was built surgically, AAC + H₂S group and H₂S group were injected intraperitoneally with H₂S (100 µmol/kg) daily. The rats in the control group and the AAC group were injected with the same amount of PBS. We observed that H₂S can improve left ventricular function and the deposition of myocardial collagen fibers, inhibit pyroptosis, down-regulate the expression of P-eif2α in myocardial tissue, and inhibit cell autophagy by activating the phosphatidylinositol 3-kinase (PI3K)/AKT1 signaling pathway (p < 0.05). In addition, angiotensin II (1 µM) H9c2 cardiomyocytes were injured in vitro experiments, and it was also observed that pyroptosis was inhibited after H₂S (400 µmol/kg) intervention, the expression of P-eif2α in cardiomyocytes was significantly down-regulated, and the PI3K/AKT1 signaling pathway was activated at the same time. Therefore, increasing the expression of P-eif2α reverses the activation of the PI3K/AKT1 signaling pathway by H₂S. In conclusion, these findings suggest that exogenous H₂S can ameliorate MF in rats with AAC by inhibiting pyroptosis, and the mechanism may be associated with inhibiting the phosphorylation of eif2α and activating the PI3K/AKT1 signaling pathway to inhibit excessive cell autophagy.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2020년도 추계국제학술대회
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• pp.56-56
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• 2020

감초는 다년생 콩과(Leguminocae) 식물로 국내에서 시중가격이 높은 만주감초가 일부 재배되고 있다. 우리나라에서 감초 재배법이 불완전한 상황에서 한반도의 기후변화에 의한 온도 상승은 약용작물의 생산 및 품질에 많은 영향을 미칠 것으로 예상되므로 본 연구에서는 재배환경 중 온도 조건만 조절할 수 있는 온도구배터널(temperature gradient tunnel system)을 이용하여 4개의 T1(외기온도+0.5~1.3℃), T2(+1.3~2.2℃), T3(+2.2~3.2℃), T4(+3.2~4.0℃) 처리로 온도구배 하여 4년생 만주감초(Glycyrrhiza uralensis F.)를 재배하였다. 지하부가 오래된 모주와 신초1의 경우 저온(T1)과 중간구간(T2, T3)에서 초장과 총화수가 우세하였고, 번식이 가장 늦은 신초2의 경우 중간구간(T2, T3)에서의 생육 및 개화반응이 뚜렷했다. 각 온도처리구마다 3개의 감초 개체를 선발하여 모주의 정단으로부터 5개의 성엽을 채취하였다. Reverse transcription quantitative PCR (RT-qPCR)은 AccuPower® GreenStar^TM RT-qPCR Master Mix (Bioneer, Korea)를 이용하여 진행되었다. Primer 디자인은 NCBI Primer-blast 프로그램을 사용해 제작하였고 ABI StepOne real time system (Applied Biosystem)의 melting curve analysis에서 one-peak test를 통해 gene specific primer임을 확인하였다. 각 온도처리구의 감초 잎에서 RNA를 추출하였고, RT-qPCR을 통해 감초의 유전자 발현양상을 비교, 분석하였다. Phytochrome interacting factor 4 (PIF4)는 식물 호르몬을 유발하는 전사조절을 조정함으로써 고온 신호전달에 핵심적인 역할을 수행한다. 활성화된 Phytochrome B(PhyB)는 PIF4의 활성을 억제한다고 알려졌다. Eukaryotic initiation factors(eIFs)는 mRNA 번역 개시인자로 유전자 발현과 특정 단백질 생산을 조절하는 역할을 한다. 본 결과는 온도조건에서 반응하는 생리적 변화를 전사체 수준에서 조사 분석하여 생리해석의 기초자료로 활용, 국내 감초 재배를 위한 환경조건 구명 및 적지 선정 기초자료로서 활용을 기대한다.

• The Plant Pathology Journal
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• 제37권1호
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• pp.47-56
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• 2021

Plants protect against viruses through passive and active resistance mechanisms, and in most cases characterized thus far, natural recessive resistance to potyviruses has been mapped to mutations in the eukaryotic initiation factor eIF4E or eIF(iso)4E genes. Five eIF4E copies and three eIF(iso)4E copies were detected in Brassica rapa. The eIF4E and eIF(iso)4E genes could interact with turnip mosaic virus (TuMV) viral protein linked to the genome (VPg) to initiate virus translation. From the yeast two-hybrid system (Y2H) and bimolecular fluorescence complementation (BiFC) assays, the TuMV-CHN2/CHN3 VPgs could not interact with BraA.eIF4E.a/c or BraA.eIF(iso)4E.c, but they could interact with BraA.eIF(iso)4E.a in B. rapa. Further analysis indicated that the amino acid substitution L186F (nt T556C) in TuMV-UK1 VPg was important for the interaction networks between the TuMV VPg and eIF(iso)4E proteins. An interaction model of the BraA. eIF(iso)4E protein with TuMV VPg was constructed to infer the effect of the significant amino acids on the interaction of TuMV VPgs-eIF(iso)4Es, particularly whether the L186F in TuMV-UK1 VPg could change the structure of the TuMV-UK1 VPg protein, which may terminate the interaction of the BraA.eIF(iso)4E and TuMV VPg protein. This study provides new insights into the interactions between plant viruses and translation initiation factors to reveal the working of key amino acids.

• Journal of Animal Science and Technology
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• 제64권3호
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• pp.481-499
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• 2022

This study aims to determine the effects of D-methionine (D-Met) isomer and the methionine precursor 2-hydroxy-4-methylthiobutanoic acid i (HMBi) supplementation on milk protein synthesis on immortalized bovine mammary epithelial cell (MAC-T). MAC-T cells were seeded using 10-cm dishes and cultured in Dulbecco's modified Eagle's medium/F12 (DMEM/F12) basic medium. The basic medium of DMEM/F12 was replaced with the lactogenic DMEM/ F12 differentiation medium when 90% of MAC-T cells reached confluency. The best dosage at 0.6 mM of D-Met and HMBi and incubation time at 72 h were used uniformly for all treatments. Each treatment was replicated six times wherein treatments were randomly assigned in a 6-well plate. Cell, medium, and total protein were determined using a bicinchoninic acid protein assay kit. Genes, proteomics and metabolomics analyses were also done to determine the mechanism of the milk protein synthesis pathway. Data were analyzed by two-way analysis of variance (ANOVA) with supplement type and plate as fixed effects. The least significant difference test was used to evaluate the differences among treatments. The HMBi treatment group had the highest beta-casein and S6 kinase beta-1 (S6K1) mRNA gene expression levels. HMBi and D-Met treatments have higher gene expressions compared to the control group. In terms of medium protein content, HMBi had a higher medium protein quantity than the control although not significantly different from the D-Met group. HMBi supplementation stimulated the production of eukaryotic translation initiation factor 3 subunit protein essential for protein translation initiation resulting in higher medium protein synthesis in the HMBi group than in the control group. The protein pathway analysis results showed that the D-Met group stimulated fructose-galactose metabolism, glycolysis pathway, phosphoinositide 3 kinase, and pyruvate metabolism. The HMBi group stimulated the pentose phosphate and glycolysis pathways. Metabolite analysis revealed that the D-Met treatment group increased seven metabolites and decreased uridine monophosphate (UMP) production. HMBi supplementation increased the production of three metabolites and decreased UMP and N-acetyl-L-glutamate production. Taken together, D-Met and HMBi supplementation are effective in stimulating milk protein synthesis in MAC-T cells by genes, proteins, and metabolites stimulation linked to milk protein synthesis.

• Nutrition Research and Practice
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• 제11권6호
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• pp.445-451
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• 2017

BACKGROUND/OBJECTIVES: Endoplasmic reticulum (ER) stress is positively associated with atherosclerosis via elevating macrophage cell death and plaque formation, in which oxidative stress plays a pivotal role. Antioxidative, lipid-lowering, and anti-atherogenic effects of kimchi, a Korean fermented vegetable, have been established, wherein capsaicin, ascorbic acid, quercetin, 3-(4'-hydroxyl-3',5'-dimethoxyphenyl)propionic acid, and lactic acids were identified. In this study, mechanisms of action of kimchi methanol extracts (KME) on fatty streak formation via suppression of ER stress and apoptosis in aorta were examined in low-density lipoprotein receptor knockout mice. MATERIALS AND METHODS: Mice fed a high cholesterol diet with an oral administration of KME (KME group, $200 mg{\cdot}kg-bw^{-1}{\cdot}day^{-1}$) or distilled water (control group) for 8 weeks (n = 20 for group). Plasma lipid and oxidative stress levels were evaluated. Protein expression was measured by western blot assay. Fatty streak lesion size and the degree of apoptosis were examined in the aorta. RESULTS: Compared to the control group, in the KME group, plasma lipids levels were decreased and oxidative stress was alleviated (P < 0.05). Protein expression levels of nuclear factor (erythroid-derived 2)-like 2-mediated antioxidants in aorta were increased whereas those for ER stress markers, glucose regulated protein 78, phospho-protein kinase RNA-like ER kinase, phospho-eukaryotic initiation factor 2 subunit ${\alpha}$, X-box binding protein 1, and C/EBP homologous protein were decreased in the KME group (P < 0.05). Moreover, apoptosis was suppressed via downregulation of phospho-c-Jun N-terminal kinase, bcl-2-associated X protein, caspases-9, and -3 with a concomitant upregulation of anti-apoptotic protein, B-cell lymphoma 2 (P < 0.05). Fatty streak lesion size was reduced and the degree of apoptosis was less severe in the KME group (P < 0.05). CONCLUSIONS: In conclusion, antioxidant activity of KME might prevent fatty streak formation through, in part, inhibition of ER stress and apoptosis in aortic sinus where macrophages are harbored.

• Parasites, Hosts and Diseases
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• 제50권3호
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• pp.243-247
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• 2012

Ascaris suum eggs are inactivated by composting conditions; however, it is difficult to find functional changes in heat-treated A. suum eggs. Here, unembryonated A. suum eggs were incubated at $20^{\circ}C$, $50^{\circ}C$, and $70^{\circ}C$ in vitro, and the gene expression levels related to viability, such as eukaryotic translation initiation factor 4E (IF4E), phosphofructokinase 1 (PFK1), and thioredoxin 1 (TRX1), and to apoptosis, such as apoptosis-inducing factor 1 (AIF1) and cell death protein 6 (CDP6), were evaluated by real-time quantitative RT-PCR. No prominent morphological alterations were noted in the eggs at $20^{\circ}C$ until day 10. In contrast, the eggs developed rapidly, and embryonated eggs and hatched larvae began to die, starting on day 2 at $50^{\circ}C$ and day 1 at $70^{\circ}C$. At $20^{\circ}C$, IF4E, PFK1, and TRX1 mRNA expression was significantly increased from days 2-4; however, AIF1 and CDP6 mRNA expression was not changed significantly. IF4E, PFK1, and TRX1 mRNA expression was markedly decreased from day 2 at $50^{\circ}C$ and $70^{\circ}C$, whereas AIF1 and CDP6 mRNA expression was significantly increased. The expressions of HSP70 and HSP90 were detected for 9-10 days at $20^{\circ}C$, for 3-5 days at $50^{\circ}C$, and for 2 days at $70^{\circ}C$. Taken together, incremental heat increases were associated with the rapid development of A. suum eggs, decreased expression of genes related to viability, and earlier expression of apoptosis-related genes, and finally these changes of viability- and apoptosis-related genes of A. suum eggs were associated with survival of the eggs under temperature stress.

• Journal of Nutrition and Health
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• 제53권6호
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• pp.583-595
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• 2020

본 연구에서는 괭생이 모자반 추출물의 소포체 스트레스 억제 효능을 연구하기 위하여 HepG2 간세포에 PA를 처리하여 소포체 스트레스를 유발한 후 추출물을 처리하여 UPR 관련 인자 발현 정도를 측정하였다. PA 750 μM 처리 시 UPR 관련 인자 (p-IRE1α, p-eIF2α, CHOP)의 단백질 발현이 가장 높게 나타나 소포체 스트레스를 효과적으로 유도함을 확인하였고 PA 750 μM를 12시간 처리 시 UPR 관련 인자 (p-IRE1α, p-eIF2α, CHOP)의 단백질 발현이 가장 높음을 확인하였다. 괭생이 모자반 처리 시 PA에 의해 상향 조절된 UPR 관련 인자의 mRNA 및 단백질 발현이 감소하여 PA로 유도된 소포체 스트레스에 대한 억제 효능이 있음을 보여주었다. 또한, 괭생이 모자반은 SIRT2, SIRT6 및 SIRT7의 mRNA의 발현을 증가시킴으로써 괭생이 모자반의 소포체 스트레스 억제 효능이 SIRT에 의한 것으로 확인되었다. 이러한 결과는 괭생이 모자반이 다양한 소포체 스트레스 관련 질병의 예방과 치료에 활용가능성이 있음을 시사한다.

• Animal Bioscience
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• 제36권6호
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• pp.851-860
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• 2023

Objective: This study aims to evaluate the target genes of gga-miR-20a-5p and the regulated immune responses in the chicken macrophage cell line, HD11, by the exosome-mediated delivery of miR-20a-5p. Methods: Exosomes were purified from the chicken macrophage cell line HD11. Then, mimic gga-miR-20p or negative control miRNA were internalized into HD11 exosomes. HD11 cells were transfected with gga-miR-20a-5p or negative control miRNA containing exosomes. After 44 h of transfection, cells were incubated with or without 5 ㎍/mL poly(I:C) for 4 h. Then, expression of target genes and cytokines was evaluated by quantitative realtime polymerase chain reaction. Results: Using a luciferase reporter assay, we identified that gga-miR-20a-5p directly targeted interferon gamma receptor 2 (IFNGR2), mitogen-activated protein kinase 1 (MAPK1), mitogen-activated protein kinase kinase kinase 5 (MAP3K5), and mitogen-activated protein kinase kinase kinase 14 (MAP3K14). Moreover, the exosome-mediated delivery of gga-miR-20a-5p successfully repressed the expression of IFNGR2, MAPK1, MAP3K5, and MAP3K14 in HD11 cells. The expressions of interferon-stimulated genes (MX dynamin like GTPase 1 [MX1], eukaryotic translation initiation factor 2A [EIF2A], and oligoadenylate synthase-like [OASL]) and proinflammatory cytokines (interferon-gamma [IFNG], interleukin-1 beta [IL1B], and tumor necrosis factor-alpha [TNFA]) were also downregulated by exosomal miR-20a-5p. In addition, the proliferation of HD11 cells was increased by exosomal miR-20a-5p. Conclusion: The exosome-mediated delivery of gga-miR-20a-5p regulated immune responses by controlling the MAPK and apoptotic signaling pathways. Furthermore, we expected that exosomal miR-20a-5p could maintain immune homeostasis against highly pathogenic avian influenza virus H5N1 infection by regulating the expression of proinflammatory cytokines and cell death.