• Title/Summary/Keyword: C.L.T.

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Crystal Structure of Cytochrome cL from the Aquatic Methylotrophic Bacterium Methylophaga aminisulfidivorans MPT

  • Ghosh, Suparna;Dhanasingh, Immanuel;Ryu, Jaewon;Kim, Si Wouk;Lee, Sung Haeng
    • Journal of Microbiology and Biotechnology
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    • v.30 no.8
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    • pp.1261-1271
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    • 2020
  • Cytochrome cL (CytcL) is an essential protein in the process of methanol oxidation in methylotrophs. It receives an electron from the pyrroloquinoline quinone (PQQ) cofactor of methanol dehydrogenase (MDH) to produce formaldehyde. The direct electron transfer mechanism between CytcL and MDH remains unknown due to the lack of structural information. To help gain a better understanding of the mechanism, we determined the first crystal structure of heme c containing CytcL from the aquatic methylotrophic bacterium Methylophaga aminisulfidivorans MPT at 2.13 Å resolution. The crystal structure of Ma-CytcL revealed its unique features compared to those of the terrestrial homologues. Apart from Fe in heme, three additional metal ion binding sites for Na+, Ca+, and Fe2+ were found, wherein the ions mostly formed coordination bonds with the amino acid residues on the loop (G93-Y111) that interacts with heme. Therefore, these ions seemed to enhance the stability of heme insertion by increasing the loop's steadiness. The basic N-terminal end, together with helix α4 and loop (G126 to Y136), contributed positive charge to the region. In contrast, the acidic C-terminal end provided a negatively charged surface, yielding several electrostatic contact points with partner proteins for electron transfer. These exceptional features of Ma-CytcL, along with the structural information of MDH, led us to hypothesize the need for an adapter protein bridging MDH to CytcL within appropriate proximity for electron transfer. With this knowledge in mind, the methanol oxidation complex reconstitution in vitro could be utilized to produce metabolic intermediates at the industry level.

Protective Effect of Processed Saengmaek-san(SM) on Cell Damage in UV-exposed HaCaT Cell (생맥산(生脈散)이 자외선에 의한 피부각질세포의 상해에 미치는 영향)

  • Kim, Eun-Seop;Yoo, Dong-Youl
    • The Journal of Korean Obstetrics and Gynecology
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    • v.24 no.2
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    • pp.33-51
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    • 2011
  • Objectives: This study was performed to assess the protective effect of Saengmaek-san (SM) on UVB-induced HaCaT cell damage. Methods: The protective effects of Saengmaek-san(SM) were determined by UVB-induced HaCaT assay. We assessed protective effects of Saengmaek-san (SM) on LDH release and nitrite release from HaCaT. And COX-2, Bcl-2, Bax, $TNF{\alpha}$, c-jun, c-fos, NF-kB, iNOS, Bcl-xL gene expression were determined in HaCaT using real-time PCR method. Results: 1. SM inhibited LDH-release, nitrite production in UVB-exposed HaCaT. 2. SM suppressed the gene expression of COX-2, $TNF{\alpha}$ in UVB-exposed HaCaT. 3. SM increased the gene expression of Bcl-2, Bax, Bcl-xL family protein in UVB-exposed HaCaT. 4. SM suppressed the gene expression of c-jun, c-fos, NF-kB in UVB-exposed HaCaT. Conclusions: The study showed SM inhibited the cell damage in UVB-exposed HaCaT.

T-Shaped DGS for CPW (CPW에 삽입된 T자형 DGS)

  • Kim Hyung-Mi;Lee Bom-Son
    • The Journal of Korean Institute of Electromagnetic Engineering and Science
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    • v.15 no.10 s.89
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    • pp.938-943
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    • 2004
  • In this paper, we propose a T-shaped DGS for CPW and provide the closed-form solutions for power loss rate, and bandwidth of the DGS. The proposed T-shaped DGS structure has a range of capacitance(C) 5.5 times wider than the dumbbell-shaped DGS structure. In addition we also analyze relations between R, L, C values of the DGS equivalent circuit and total loss rate, BW, $\omega_0$ of the DGS.

Betulinic Acid Stimulates Glucose Uptake through the Activation of PI3K and AMPK in 3T3-L1 Adipocytes (Betulinic acid의 PI3K와 AMPK경로 활성화를 통한 3T3-L1 지방세포에서 포도당 흡수 촉진 효과)

  • Lee, Jung Kyung;Park, Jae Eun;Han, Ji Sook
    • Journal of Life Science
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    • v.32 no.10
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    • pp.762-770
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    • 2022
  • Hyperglycemia in type 2 diabetes can be alleviated by promoting cellular glucose uptake. Betulinic acid (3β,-3-hydroxy-lup-20(29)-en-28-oic acid) is a pentacyclic lupane-type triterpenoid compound. Although there have been studies on the antidiabetic activity of betulinic acid, studies on cellular glucose uptake are lacking. We investigated the effects of betulinic acid on glucose uptake and its mechanism of action in 3T3-L1 adipocytes. Betulinic acid significantly stimulated glucose uptake in 3T3-L1 adipocytes by increasing the phosphorylation of the insulin receptor substrate 1-tyrosine (IRS-1tyr) in the insulin signaling pathway, which in turn stimulated the activation of phosphoinositide 3-kinase (PI3K) and the phosphorylation of protein kinase B (Akt). The activation of PI3K and Akt by betulinic acid translocated glucose transporter 4 to the plasma membrane (PM-GLUT4), thereby increasing the expression of PM-GLUT4 and thus stimulating cellular glucose uptake. Betulinic acid also significantly increased the phosphorylation/activation of AMP-activated protein kinase (AMPK) and acetyl-CoA carboxylase. The activation of PI3K and AMPK by betulinic acid was confirmed using the PI3K inhibitor wortmannin and the AMPK inhibitor compound C. The increase in glucose uptake induced by betulinic acid was significantly decreased by wortmannin and compound C in the 3T3-L1 adipocytes. These results suggest that betulinic acid stimulates glucose uptake by activating PI3K and AMPK in 3T3-L1 adipocytes.

Characteristics of Flame Hardening Process for 12Cr Steels (12Cr 강의 이동 화염경화 공정 특성)

  • Kim Gwang-Ho;Lee Min-Ku;Kim Kyeong-Ho;Kim Whung-Whoe;Rhee Chang-Kyu;Kim Gil-Mu
    • Journal of the Korean institute of surface engineering
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    • v.39 no.2
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    • pp.49-56
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    • 2006
  • In this study, the movable flame hardening process of 12Cr steel for a uniform hardness and desirable residual stress have been investigated. For this, the temperature cycles have been controlled accurately as a function of the three processing variables, the flame intensity $I_f$, the scanning velocity $V_s$, and the initial flame holding time $t_h$, where the standard surface temperature $T_{s,\;max}$, was maintained at $960^{\circ}C$. The optimized conditions were $V_s=0.68mn/s\;and\;t_h=67sec$ for the $C_3H_8:O_2\;=\;5:20l/min,\;V_s=0.80mm/s$ and $t_h=56sec$ for the $C_3H_8:O_2=6:24l/min,\;V_s=1.01mm/s\;and\;t_h=48sec$ for the $C_3H_8:O_2=7:28l/min,\;and\;V_s=1.15mm/s$ and $t_h=39sec$ for the $C_3H_8:O_2$=8:32 l/min. The optimally flame-hardened surface exhibited uniform distributions of the hardness and residual compressive stress over the treated area with moderate levels of $470{\sim}490HV_{0.2}$in hardness and $-300{\sim}-450MPa$ in residual stress, which were acceptable on the basis of the acceptance criteria of Siemens AG-KWU and GE Power Generation Engineering.

Red pepper seed water extract inhibits preadipocyte differentiation and induces mature adipocyte apoptosis in 3T3-L1 cells

  • Kim, Hwa-Jin;You, Mi-Kyoung;Lee, Young-Hyun;Kim, Hyun-Jung;Adhikari, Deepak;Kim, Hyeon-A
    • Nutrition Research and Practice
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    • v.12 no.6
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    • pp.494-502
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    • 2018
  • BACKGROUND/OBJECTIVES: Reducing the number of adipocytes by inducing apoptosis of mature adipocytes as well as suppressing differentiation of preadipocytes plays an important role in preventing obesity. This study examines the anti-adipogenic and pro-apoptotic effect of red pepper seed water extract (RPS) prepared at $4^{\circ}C$ (RPS4) in 3T3-L1 cells. MATERIALS/METHODS: Effect of RPS4 or its fractions on lipid accumulation was determined in 3T3-L1 cells using oil red O (ORO) staining. The expressions of AMP-activated protein kinase (AMPK) and adipogenic associated proteins [peroxisome proliferator-activated receptor-${\gamma}$ ($PPAR-{\gamma}$), CCAAT/enhancer-binding proteins ${\alpha}$ (C/EBP ${\alpha}$), sterol regulatory element binding protein-1c (SREBP-1c), fatty acid synthase (FAS), and acetyl-CoA carboxylase (ACC)] were measured in 3T3-L1 cells treated with RPS4. Apoptosis and the expression of Akt and Bcl-2 family proteins [B-cell lymphoma 2 (Bcl-2), Bcl-2-associated death promoter (Bad), Bcl-2 like protein 4 (Bax), Bal-2 homologous antagonist/killer (Bak)] were measured in mature 3T3-L1 cells treated with RPS4. RESULTS: Treatment of RPS4 ($0-75{\mu}g/mL$) or its fractions ($0-50{\mu}g/mL$) for 24 h did not have an apparent cytotoxicity on pre and mature 3T3-L1 cells. RPS4 significantly suppressed differentiation and cellular lipid accumulation by increasing the phosphorylation of AMPK and reducing the expression of $PPAR-{\gamma}$, C/EBP ${\alpha}$, SREBP-1c, FAS, and ACC. In addition, all fractions except ethyl acetate fraction significantly suppressed cellular lipid accumulation. RPS4 induced the apoptosis of mature adipocytes by hypophosphorylating Akt, increasing the expression of the pro-apoptotic proteins, Bak, Bax, and Bad, and reducing the expression of the anti-apoptotic proteins, Bcl-2 and p-Bad. CONCLUSIONS: These finding suggest that RPS4 can reduce the numbers as well as the size of adipocytes and might useful for preventing and treating obesity.

Growing pigs developed different types of diabetes induced by streptozotocin depending on their transcription factor 7-like 2 gene polymorphisms

  • Tu, Ching-Fu;Hsu, Chi-Yun;Lee, Meng-Hwan;Jiang, Bo-Hui;Guo, Shyh-Forng;Lin, Chai-Ching;Yang, Tien-Shuh
    • Laboraroty Animal Research
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    • v.34 no.4
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    • pp.185-194
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    • 2018
  • The different polymorphisms of the transcription factor 7-like 2 (TCF7L2) gene promote variances in diabetes susceptibility in humans. We investigated whether these genotypes also promote differences in diabetic susceptibility in commercial pigs. Growing pigs (Landrace, both sex, 50-60 kg) with the C/C (n=4) and T/T (n=5) TCF7L2 genotypes were identified and intravenously injected with streptozotocin (STZ, 40 mg/kg) twice in weekly intervals, then a high-energy diet was offered. Oral glucose tolerance tests, blood analyses and the homeostasis model assessment-insulin resistance (HOMA-IR) index calculations were performed. The animals were sacrificed at the end of 12 weeks of treatment to reveal the pancreas histomorphometry. The results showed that all of the treated pigs grew normally despite exhibiting hyperglycemia at two weeks after the induction. The glycemic level of the fasting or postprandial pigs gradually returned to normal. The fasting insulin concentration was significantly decreased for the T/T carriers but not for the C/C carriers, and the resulting HOMA-IR index was significantly increased for the C/C genotype, indicating that the models of insulin dependence and resistance were respectively developed by T/T and C/C carriers. The histopathological results illustrated a significant reduction in the pancreas mass and insulin active sites, which suggested increased damage. The results obtained here could not be compared with previous studies because the TCF7L2 background has not been reported. Growing pigs may be an excellent model for diabetic in children if the animals are genetically pre-selected.

The effect of eleutherococcus senticosus on metabolism-associated protein expression in 3T3-L1 and C2C12 cells

  • Hashimoto, Takeshi;Okada, Yoko;Yamanaka, Atsushi;Ono, Natsuhiko;Uryu, Keisuke;Maru, Isafumi
    • Korean Journal of Exercise Nutrition
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    • v.24 no.3
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    • pp.13-18
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    • 2020
  • [Purpose] In vivo studies have demonstrated the ergogenic benefits of eleutherococcus senticosus (ES) supplementation. ES has been observed to enhance endurance capacity, improve cardiovascular function, and alter metabolic functions (e.g., increased fat utilization); however, the exact mechanisms involved remain unknown. We aimed to determine whether ES could effectively induce fat loss and improve muscle metabolic profiles through increases in lipolysis- and lipid metabolism-associated protein expression in 3T3-L1 adipocytes and C2C12 skeletal muscle cells, respectively, to uncover the direct effects of ES on adipocytes and skeletal muscle cells. [Methods] Different doses of ES extracts (0.2, 0.5, and 1.0 mg/mL) were added to cells (0.2 ES, 0.5 ES, and 1.0 ES, respectively) for 72 h and compared to the vehicle control (control). [Results] The intracellular triacylglycerol (TG) content significantly decreased (p < 0.05 for 0.2 ES, p < 0.01 for 0.5 ES and 1.0 ES) in 3T3-L1 cells. Adipose triglyceride lipase, which is involved in active lipolysis, was significantly higher in the 1.0 ES group than in the control group (p < 0.01) of 3T3-L1 adipocytes. In C2C12 cells, the mitochondrial protein voltage-dependent anion channel (VDAC) was significantly increased in the 1.0 ES group (p < 0.01). Furthermore, we found that 1.0 ES activated both 5' AMP-activated protein kinase (AMPK) and acetyl-CoA carboxylase (ACC) in skeletal muscle cells (p < 0.01). [Conclusion] These findings suggest that ES extracts decreased TG content, presumably by increasing lipase in adipocytes and metabolism-associated protein expression as well as mitochondrial biogenesis in muscle cells. These effects may corroborate previous in vivo findings regarding the ergogenic effects of ES supplementation.