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Robinetin Alleviates Metabolic Failure in Liver through Suppression of p300-CD38 Axis

  • Ji-Hye Song;Hyo-Jin Kim;Jangho Lee;Seung-Pyo Hong;Min-Yu Chung;Yu-Geun Lee;Jae Ho Park;Hyo-Kyoung Choi;Jin-Taek Hwang
    • Biomolecules & Therapeutics
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    • v.32 no.2
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    • pp.214-223
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    • 2024
  • Metabolic abnormalities in the liver are closely associated with diverse metabolic diseases such as non-alcoholic fatty liver disease, type 2 diabetes, and obesity. The aim of this study was to evaluate the ameliorating effect of robinetin (RBN) on the significant pathogenic features of metabolic failure in the liver and to identify the underlying molecular mechanism. RBN significantly decreased triglyceride (TG) accumulation by downregulating lipogenesis-related transcription factors in AML-12 murine hepatocyte cell line. In addition, mice fed with Western diet (WD) containing 0.025% or 0.05% RBN showed reduced liver mass and lipid droplet size, as well as improved plasma insulin levels and homeostatic model assessment of insulin resistance (HOMA-IR) values. CD38 was identified as a target of RBN using the BioAssay database, and its expression was increased in OPA-treated AML-12 cells and liver tissues of WD-fed mice. Furthermore, RBN elicited these effects through its anti-histone acetyltransferase (HAT) activity. Computational simulation revealed that RBN can dock into the HAT domain pocket of p300, a histone acetyltransferase, which leads to the abrogation of its catalytic activity. Additionally, knock-down of p300 using siRNA reduced CD38 expression. The chromatin immunoprecipitation (ChIP) assay showed that p300 occupancy on the promoter region of CD38 was significantly decreased, and H3K9 acetylation levels were diminished in lipid-accumulated AML-12 cells treated with RBN. RBN improves the pathogenic features of metabolic failure by suppressing the p300-CD38 axis through its anti-HAT activity, which suggests that RBN can be used as a new phytoceutical candidate for preventing or improving this condition.

An analysis of elementary students' reasoning on the sum of triangle angles ('삼각형 세 각의 크기의 합'에 관한 초등학생의 추론 연구)

  • Kim, Ji Hyun
    • Education of Primary School Mathematics
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    • v.27 no.2
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    • pp.155-171
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    • 2024
  • This study compared and analyzed students' reasoning processes and justification methods when introducing the concept of "the sum of angles in a triangle" in mathematics classes with a focus on both measurement and geometric aspects. To confirm this, the research was conducted in a 4th-grade class at H Elementary School in Suwon, Gyeonggi-do, South Korea. The conclusions drawn from this study are as follows. First, there is a significant difference when introducing "the sum of angles in a triangle" in mathematics classes from a measurement perspective compared to a geometric perspective. Second, justifying the statement "the sum of angles in a triangle is 180°" is more effective when explained through a measurement approach, such as "adding the sizes of the three angles gives 180°," rather than a geometric approach, such as "the sum of the angles forms a straight angle." Since elementary students understand mathematical knowledge through manipulative activities, the level of activity is connected to the quality of mathematics learning. Research on this reasoning process will serve as foundational material for approaching the concept of "the sum of angles in a triangle" within the "Geometry and Measurement" domain of the Revised 2022 curriculum.

Suppression of EGFR/STAT3 activity by lupeol contributes to the induction of the apoptosis of human non-small cell lung cancer cells

  • TAE‑RIN MIN;HYUN‑JI PARK;KI‑TAE HA;GYOO‑YONG CHI;YUNG‑HYUN CHOI;SHIN‑HYUNG PARK
    • International Journal of Oncology
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    • v.55 no.1
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    • pp.320-330
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    • 2019
  • The aim of this study was to investigate the underlying mechanisms responsible for the anticancer effects of lupeol on human non-small cell lung cancer (NSCLC). MTT assay and Trypan blue exclusion assay were used to evaluate the cell viability. DAPI staining and flow cytometric analysis were used to detect apoptosis. Molecular docking and western blot analysis were performed to determine the target of lupeol. We found that lupeol suppressed the proliferation and colony formation of NSCLC cells in a dose-dependent manner. In addition, lupeol increased chromatin condensation, poly(ADP-ribose) polymerase (PARP) cleavage, sub-G1 cell populations, and the proportion of Annexin V-positive cells, indicating that lupeol triggered the apoptosis of NSCLC cells. Notably, lupeol inhibited the phosphorylation of epithelial growth factor receptor (EGFR). A docking experiment revealed that lupeol directly bound to the tyrosine kinase domain of EGFR. We observed that the signal transducer and activator of transcription 3 (STAT3), a downstream molecule of EGFR, was also dephosphorylated by lupeol. Lupeol suppressed the nuclear translocation and transcriptional activity of STAT3 and downregulated the expression of STAT3 target genes. The constitutive activation of STAT3 by STAT3 Y705D overexpression suppressed lupeol-induced apoptosis, demonstrating that the inhibition of STAT3 activity contributed to the induction of apoptosis. The anticancer effects of lupeol were consistently observed in EGFR tyrosine kinase inhibitor (TKI)-resistant H1975 cells (EGFR L858R/T790M). Taken together, the findings of this study suggest that lupeol may be used, not only for EGFR TKI-naïve NSCLC, but also for advanced NSCLC with acquired resistance to EGFR TKIs.

A Novel Truncated CHAP Modular Endolysin, CHAPSAP26-161, That Lyses Staphylococcus aureus, Acinetobacter baumannii, and Clostridioides difficile, and Exhibits Therapeutic Effects in a Mouse Model of A. baumannii Infection

  • Yoon-Jung Choi;Shukho Kim;Ram Hari Dahal;Jungmin Kim
    • Journal of Microbiology and Biotechnology
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    • v.34 no.8
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    • pp.1718-1726
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    • 2024
  • Development of novel antibacterial agents is imperative due to the increasing threat of antibiotic-resistant pathogens. This study aimed to develop the enhanced antibacterial activity and in-vivo efficacy of a novel truncated endolysin, CHAPSAP26-161, derived from the endolysin LysSAP26, against multidrug-resistant bacteria. CHAPSAP26-161 exhibited higher protein purification efficiency in E. coli and antibacterial activity than LysSAP26. Moreover, CHAPSAP26-161 showed the higher lytic activity against A. baumannii with minimal bactericidal concentrations (MBCs) of 5-10 ㎍/ml, followed by Staphylococcus aureus with MBCs of 10-25 ㎍/ml. Interestingly, CHAPSAP26-161 could lyse anaerobic bacteria, such as Clostridioides difficile, with MBCs of 25-50 ㎍/ml. At pH 4-8 and temperatures of 4℃-45℃, CHAPSAP26-161 maintained antibacterial activity without remarkable difference. The lytic activity of CHAPSAP26-161 was increased with Zn2+. In vivo tests demonstrated the therapeutic effects of CHAPSAP26-161 in murine systemic A. baumannii infection model. In conclusion, CHAPSAP26-161, a truncated endolysin that retains only the CHAP domain from LysSAP26, demonstrated enhanced protein purification efficiency and antibacterial activity compared to LysSAP26. It further displayed broad-spectrum antibacterial effects against S. aureus, A. baumannii, and C. difficile. Our in vitro and in-vivo results of CHAPSAP26-161 highlights its promise as an innovative therapeutic option against those bacteria with multiple antibiotic resistance.

The Effect of Insulin-Like Growth Factor-I(IGF-I) and IGF Binding Protein-3(IGFBP-3) on Cellular Proliferation in Mouse 3T3 Fibroblast Cells (마우스 섬유아세포(3T3 fibroblast cells)에서 Insulin-like Growth Factor-I(IGF-I) 및 IGF Binding Protein-3 (IGFBP-3)이 세포증식에 미치는 영향)

  • Cho, Chul-Ho;Kwak, Seung-Min;Moon, Tae-Hun;Cho, Jae-Hwa;Ryu, Jeong-Seon;Lee, Hyong-Lyeol
    • Tuberculosis and Respiratory Diseases
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    • v.47 no.5
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    • pp.618-628
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    • 1999
  • Background: Cell growth is a balance between cell proliferation and cell death. Insulin-like growth factor-I(IGF-I), which binds IGF-I receptor(IGF-IR), mediates cellular proliferation as a potent mitogen. IGF binding protein-3(IGFBP-3) as a circulating major IGFBP can inhibit or enhance the effects of IGF-I on cellular growth by binding IGFs. Methods: We investigated the expressions of mRNA of IGF-I and IGF-IR by northern blot and phosphorylation of IGF-IR with the treatment of IGF-I by western blot in 3T3 fibroblast cells. The cellular proliferations of 3T3 cells with the treatments of IGF-I were evaluated using $^3H$-thymidine incorporation and MTT assay. Also to observe the effect of IGFBP-3 on cellular proliferation, 3T3 cells were treated with anti-IGFBP-3 and ${\alpha}IR_3$(monoclonal antibody to IGF-IR) alone or in combination. Results: Our results demonstrated that 3T3 cells showed mRNA expressions of IGF-I and IGF-IR and the IGF-I increased phosphorylation of IGF-IR. The treatments of 3T3 cells with IGF-I increased cellular proliferation in 5 % and 1 % seruma-containing media, not in serum-free media. The addition of anti-IGFBP-3 to neutralize IGFBP-3 showed 2-fold increase of cellular proliferation, and also co-incubation of anti-IGFBP-3 and ${\alpha}IR_3$ together showed similar increase of cellular proliferation in 3T3 cells. Interestingly, when the cells were pretreated with ${\alpha}IR_3$ for 4 hr, prior to the simultaneous addition of ${\alpha}IR_3$ and anti-IGFBP-3, anti-IGFBP-3-mediated cellular proliferation was decreased to control level. All of these results suggest that free IGF-I released from IGF-I/IGFBP-3 complex would be involved in the cellular proliferation. Conclusion: IGF-I is a mitogen through the activation of IGF-IR in 3T3 cells, and IGFBP-3 could be a potent inhibitor for IGF-I action by binding IGF-I.

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Study on Oxidation and Coercivity of Nd2Fe14B Compound Crystal (Nd2Fe14B 화합물 결정의 산화 및 보자력에 관한 연구)

  • Kwon, H.W.;Yu, J.H.
    • Journal of the Korean Magnetics Society
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    • v.22 no.3
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    • pp.85-90
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    • 2012
  • Oxidation of the $Nd_2Fe_{14}B$ compound crystal and its effect on the coercivity of the fine $Nd_2Fe_{14}B$ crystal particles were investigated. Oxidation kinetics of the $Nd_2Fe_{14}B$ compound crystal was investigated using an excessively grown $Nd_2Fe_{14}B$ grains in the $Nd_{15}Fe_{77}B_8$ alloy ingot. Oxidation of the $Nd_2Fe_{14}B$ compound crystal occurred by dissociation of the phase into multi-phase mixture of ${\alpha}$-Fe, $Fe_3B$, and Nd oxides. Oxidation rate of the $Nd_2Fe_{14}B$ compound crystal showed no dependence on the crystallographic direction. The oxidation reaction was modeled according to simple linear relationship. Activation energy for the oxidation of $Nd_2Fe_{14}B$ compound crystal was calculated to be approximately 26.8 kJ/mol. Fine $Nd_2Fe_{14}B$ crystal particles in near single domain size was prepared by ball milling of the HDDR-treated $Nd_{15}Fe_{77}B_8$ alloy, and these particles were used for investigating the effect of oxidation on the coercvity. The near single domain size $Nd_2Fe_{14}B$ crystal particles (${\fallingdotseq}0.3\;{\mu}m$) had high coercivity over 9 kOe. However, the coercivity was radically reduced as the temperature increased in air (<2 kOe at $200^{\circ}C$). This radical coercivity reduction was attributed to the soft magnetic phases, ${\alpha}$-Fe and $Fe_3B$, which were formed on the surface of the fine particles due to the oxidation.

Thickness Dependence of Amorphous CoSiB/Pd Multilayer with Perpendicular Magnetic Anisotropy (비정질 강자성체 CoSiB/Pd 다층박막의 두께에 따른 수직자기이방성 변화)

  • Yim, H.I.
    • Journal of the Korean Magnetics Society
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    • v.23 no.4
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    • pp.122-125
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    • 2013
  • Perpendicular magnetic anisotropy (PMA) is the phenomenon of magnetic thin film which is preferentially magnetized in a direction perpendicular to the film's plane. Amorphous multilayer with PMA has been studied as the good candidate to realization of high density STT-MRAM (Spin Transfer Torque-Magnetic Random Access Memory). The current issue of high density STT-MRAM is a decrease in the switching current of the device and an application of amorphous materials which are most suitable devices. The amorphous ferromagnetic material has low saturated magnetization, low coercivity and high thermal stability. In this study, we presented amorphous ferromagnetic multilayer that consists of an amorphous alloy CoSiB and a nonmagnetic material Pd. We investigated the change of PMA of the $[CoSiB\;t_{CoSiB}/Pd\;1.3nm]_5$ multilayer ($t_{CoSiB}$ = 0.1, 0.2, 0.3, 0.4, 0.5, 0.6 nm, and $t_{Pd}$ = 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6 nm) and $[CoSiB\;0.3nm/Pd\;1.3nm]_n$ multilayer (n = 3, 5, 7, 9, 11, 13). This multilayer is measured by VSM (Vibrating Sample Magnetometer) and analyzed magnetic properties like a coercivity ($H_c$) and a magnetization ($M_s$). The coercivity in the $[CoSiB\;t_{CoSiB}\;nm/Pd\;1.3nm]_5$ multi-layers increased with increasing $t_{CoSiB}$ to reach a maximum at $t_{CoSiB}$ = 0.3 nm and then decreased for $t_{CoSiB}$ > 0.3 nm. The lowest saturated magnetization of $0.26emu/cm^3$ was obtained in the $[CoSiB\;0.3nm/Pd\;1.3nm]_3$ multilayer whereas the highest coercivity of 0.26 kOe was obtained in the $[CoSiB\;0.3nm/Pd\;1.3nm]_5$ mutilayer. Additional Pd layers did not contribute to the perpendicular magnetic anisotropy. The single domain structure evolved in to a striped multi-domain structure as the bilayer repetition number n was increased above 7 after which (n > 7) the hysteresis loops had a bow-tie shapes.

The Region-of-Interest Based Pixel Domain Distributed Video Coding With Low Decoding Complexity (관심 영역 기반의 픽셀 도메인 분산 비디오 부호)

  • Jung, Chun-Sung;Kim, Ung-Hwan;Jun, Dong-San;Park, Hyun-Wook;Ha, Jeong-Seok
    • Journal of the Institute of Electronics Engineers of Korea SP
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    • v.47 no.4
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    • pp.79-89
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    • 2010
  • Recently, distributed video coding (DVC) has been actively studied for low complexity video encoder. The complexity of the encoder in DVC is much simpler than that of traditional video coding schemes such as H.264/AVC, but the complexity of the decoder in DVC increases. In this paper, we propose the Region-Of-Interest (ROI) based DVC with low decoding complexity. The proposed scheme uses the ROI, the region the motion of objects is quickly moving as the input of the Wyner-Ziv (WZ) encoder instead of the whole WZ frame. In this case, the complexity of encoder and decoder is reduced, and the bite rate decreases. Experimental results show that the proposed scheme obtain 0.95 dB as the maximum PSNR gain in Hall Monitor sequence and 1.87 dB in Salesman sequence. Moreover, the complexity of encoder and decoder in the proposed scheme is significantly reduced by 73.7% and 63.3% over the traditional DVC scheme, respectively. In addition, we employ the layered belief propagation (LBP) algorithm whose decoding convergence speed is 1.73 times faster than belief propagation algorithm as the Low-Density Parity-Check (LDPC) decoder for low decoding complexity.

Cloning and Characterization of Cellulase Gene (cel5C) from Cow Rumen Metagenomic Library (소 반추위 메타게놈에서 새로운 섬유소분해효소 유전자(cel5C) 클로닝 및 유전산물의 특성)

  • Kim, Min-Keun;Barman, Dhirendra Nath;Kang, Tae-Ho;Kim, Jung-Ho;Kim, Hoon;Yun, Han-Dae
    • Journal of Life Science
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    • v.22 no.4
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    • pp.437-446
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    • 2012
  • A metagenomic library of cow rumen in the pCC1FOS phage vector was screened in $E.$ $coli$ EPI300 for cellulase activity on carboxymethyl cellulose agar plates. One clone was partially digested with $Sau$3AI, ligated into the $Bam$HI site of the pBluescript II SK+ vector, and transformed into $E.$ $coli$ $DH5{\alpha}$. We obtained a 1.5 kb insert DNA, designated $cel$5C, which hydrolyzes carboxymethyl cellulose. The cel5C gene has an open reading frame (ORF) of 1,125 bp encoding 374 amino acids. It belongs to the glycosyl hydrolase family 5 with the conserved domain LIMEGFNEIN. The molecular mass of the Cel5C protein induced from $E.$ $coli$ $DH5{\alpha}$, as analyzed by CMC SDS-PAGE, appeared to be approximately 42 kDa. The enzyme showed optimum cellulase activity at pH 4.0, and $50^{\circ}C$. We examined whether the $cel$5C gene comes from the 49 identified cow rumen bacteria using PCR. No PCR bands were identified, suggesting that the $cel$5C gene came from the unidentified cow rumen bacteria.

Construction of Mammalian Cell Expression Vector for pAcGFP-bFLIP(L) Fusion Protein and Its Expression in Follicular Granulosa Cells

  • Yang, Run Jun;Li, Wu Feng;Li, Jun Ya;Zhang, Lu Pei;Gao, Xue;Chen, Jin Bao;Xu, Shang Zhong
    • Asian-Australasian Journal of Animal Sciences
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    • v.23 no.3
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    • pp.401-409
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    • 2010
  • FLICE inhibitory protein (FLIP) is one of the important anti-apoptotic proteins in the Fas/FasL apoptotic path which has death effect domains, mimicking the pro-domain of procaspase-8. To reveal the intracellular signal transduction molecules involved in the process of follicular development in the bovine ovary, we cloned the c-FLIP(L) gene in bovine ovary tissue with the reverse transcription polymerase chain reaction (RT-PCR), deleted the termination codon in its cDNA, and directionally cloned the amplified c-FLIP(L) gene into eukaryotic expression vector pAcGFP-Nl, including AcGFP, and successfully constructed the fusion protein recombinant plasmid. After identifying by restrictive enzyme BglII/EcoRI and sequencing, pAcGFP-bFLIP(L) was then transfected into follicular granulosa cells, mediated by Lipofectamine 2000, the expression of AcGFP observed and the transcription and expression of c-FLIP(L) detected by RT-PCR and Western blot. The results showed that the cattle c-FLIP(L) was successfully cloned; the pAcGFPbFLIP(L) fusion protein recombinant plasmid was successfuly constructed by introducing a BglII/EcoRI cloning site at the two ends of the c-FLIP(L) open reading frame and inserting a Kozak sequence before the start codon. AcGFP expression was detected as early as 24 h after transfection. The percentage of AcGFP positive cells reached about 65% after 24 h. A 1,483 bp transcription was amplified by RT-PCR, and a 83 kD target protein was detected by Western blot. Construction of the pAcGFP-bFLIP(L) recombinant plasmid should be helpful for further understanding the mechanism of regulation of c-FLIP(L) on bovine oocyte formation and development.