• Journal of Microbiology and Biotechnology
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• v.15 no.4
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• pp.756-766
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• 2005

The signaling mechanism underlying aburatubolactam C-induced FasL upregulation was investigated in human Jurkat T cells. After treatment with aburatubolactam C, the src-family PTKs $p56^{lck}\;and\;p59^{fyn}$, and MAP kinases ERK2 and JNK1, were activated prior to FasL upregulation; Both $p56^{lck}\;and\;p59^{fyn}$ were directly activated 2.4- and 2.2-fold, respectively, in vitro by aburatubolactam C. The aburatubolactam C-induced cellular changes, including the activation of ERK2 and INK1, and FasL upregulation, were completely prevented by the PTK inhibitor genistein. The activation of protein kinase C (PKC$\delta,\;\epsilon\;and\;\mu$ was also induced following aburatubolactam C treatment. Although the activation of $p56^{lck}$ and tyrosine phosphorylation of the cellular proteins were not blocked by the PKC inhibitor GFl09203X, the activation of ERK2 was completely abrogated, along with a detectably enhanced JNK1 activation; FasL upregulation, and apoptosis. However, the FasL upregulation and apoptosis were significantly inhibited by the PKC activator PMA, with a remarkable increase in the ERK2 activation. The cytotoxic effect of aburatubolactam C was reduced in the presence of the anti-Fas neutralizing antibody ZB-4. Although ectopic expression of Bcl-2 failed to completely block the cytotoxicity of aburatubolactam C, it was clearly suppressed. The c-Fos mRNA expression was upregulated in a biphasic manner, where the second phasic expression overlapped with the FasL upregulation. Accordingly, these results demonstrate that aburatubolactam C-induced apoptosis is exerted, at least in part, by FasL upregulation dictated by activation of the PTK ($p56^{lck}\;and\;p59^{fyn}$) /JNKI pathway, which is negatively affected by the concurrent activation of the PKC/ERK2 pathway proximal to PTK activation.

• Proceedings of the Korean Vacuum Society Conference
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• 2011.08a
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• pp.266-267
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• 2011

질화갈륨(GaN)은 높은 전자이동도 및 높은 항복전계를 가지며 낮은 온저항으로 인하여 에너지효율이 우수하기 때문에 고출력 전력소자 분야에서 많은 관심을 받고 있다. GaN을 이용한 고출력 전력소자의 경우 상용화 수준에 근접할 만한 기술적 진보가 있었으나, 페르미 레벨 고정(Fermi-level pinning) 현상, 소자의 누설전류 등 아직 해결되어야 할 문제를 갖고 있다. 본 연구에서는 실리콘 기판 위에 성장된 GaN 에피탁시를 활용한 고출력 전력소자의 누설전류를 억제시키기 위해 오믹 접합 중 Au의 상호확산을 억제하는 중간층 금속(Mo or Ni)을 변화시켰으며 오믹 열처리 온도에 따른 특성을 비교 연구하였다. $Cl_2$와 $BCl_3$를 이용하여 0.6 ${\mu}m$ 깊이의 메사 구조가 활성영역을 형성하였고, Si 도핑된 n-GaN 위에 Ti/Al/Mo/Au (20/100/25/200 nm) 와 Ti/Al/Ni/Au (20/100/25/200 nm) 오믹 접합을 각각 설계, 제작하였다. 오믹 열처리시의 GaN 표면오염을 방지하기 위해 $SiO_2$ 희생층을 증착하였다. 오믹 접합 형성을 위해 각 750$^{\circ}C$, 800$^{\circ}C$, 850$^{\circ}C$에서 30초간 열처리를 진행 하였으며, 이후 6 : 1 BOE 용액으로 $SiO_2$ 희생층을 제거하였다. 750, 800, 850$^{\circ}C$에서 Ti/Al/Mo/Au 구조의 오믹 접합 저항은 각 2.56, 2.34, 2.22 ${\Omega}$-mm 이었으며, Ti/Al/Ni/Au 구조의 오믹 접합 저항은 각 43.72, 2.64, 1.86 ${\Omega}$-mm이었다. Isolation 누설전류를 측정하기 위해서 두 개의 오믹 접합 사이에 메사 구조가 있는 테스트 구조를 제안하였다. Isolation 누설전류는 Ti/Al/Mo/Au 구조에서 두 오믹 접합 사이의 거리가 25 ${\mu}m$이고 100 V일 때 750, 800, 850 $^{\circ}C$의 열처리 온도에서 각 1.25 nA/${\mu}m$, 2.48 nA/${\mu}m$, 8.76 nA/${\mu}m$이었으며, Ti/Al/Ni/Au 구조에서는 각 1.58 nA/${\mu}m$, 2.13 nA/${\mu}m$, 96.36 nA/${\mu}m$이었다. 열처리 온도가 증가하며 오믹 접합 저항은 감소하였으나 isolation 누설전류는 증가하였다. 750$^{\circ}C$ 열처리에서 오믹 접합 저항은Ti/Al/Mo/Au 구조가 Ti/Al/Ni/Au 구조보다 약 17배 우수하였고, 850$^{\circ}C$ 고온의 열처리 경우 Ti/Al/Mo/Au 구조의 isolation 누설전류는 8.76 nA/${\mu}m$로 Ti/Al/Ni/Au의 누설전류 96.36 nA/${\mu}m$보다 약 11배 우수하였다. Ti/Al/Mo/Au가 Ti/Al/Ni/Au 보다 오믹 접합 저항과 isolation 누설전류 측면에서 전력용 GaN 소자에 적합함을 확인하였다.

• Animal Bioscience
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• v.34 no.7
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• pp.1210-1220
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• 2021

Objective: Unlike mammals, goose fatty liver shows a strong tolerance to fatty acids without obvious injury. Stearyl-coenzyme A desaturase 1 (SCD1) serves crucial role in desaturation of saturated fatty acids (SAFs), but its role in the SAFs tolerance of goose hepatocytes has not been reported. This study was conducted to explore the role of SCD1 in regulating palmitic acid (PA) tolerance of goose primary hepatocytes. Methods: 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide was examined to reflect the effect of PA on hepatocytes viability, and quantitative polymerase chain reaction was used to detect the mRNA levels of several genes related to endoplasmic reticulum (ER) stress, inflammation, and apoptosis, and the role of SCD1 in PA tolerance of goose hepatocytes was explored using RNA interfere. Results: Our results indicated that goose hepatocytes exhibited a higher tolerant capacity to PA than human hepatic cell line (LO2 cells). In goose primary hepatocytes, the mRNA levels of fatty acid desaturation-related genes (SCD1 and fatty acid desaturase 2) and fatty acid elongate enzyme-related gene (elongase of very long chain fatty acids 6) were significantly upregulated with 0.6 mM PA treatment. However, in LO2 cells, expression of ER stress-related genes (x box-binding protein, binding immunoglobulin protein, and activating transcription factor 6), inflammatory response-related genes (interleukin-6 [IL-6], interleukin-1β [IL-1β], and interferon-γ) and apoptosis-related genes (bcl-2-associated X protein, b-cell lymphoma 2, Caspase-3, and Caspase-9) was significantly enhanced with 0.6 mM PA treatment. Additionally, small interfering RNA (siRNA) mediated downregulation of SCD1 significantly reduced the PA tolerance of goose primary hepatocytes under the treatment of 0.6 mM PA; meanwhile, the mRNA levels of inflammatory-related genes (IL-6 and IL-1β) and several key genes involved in the phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT), forkhead box O1 (FoxO1), mammalian target of rapamycin and AMPK pathways (AKT1, AKT2, FoxO1, and sirtuin 1), as well as the protein expression of cytochrome C and the apoptosis rate were upregulated. Conclusion: In conclusion, our data suggested that SCD1 was involved in enhancing the PA tolerance of goose primary hepatocytes by regulating inflammation- and apoptosis-related genes expression.

• Korean Journal of Agricultural Science
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• v.49 no.2
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• pp.175-184
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• 2022

The present study investigated the neuroprotective effects of Coreopsis lanceolate extract against hydrogen-peroxide (H₂O₂)-induced oxidative damage and cell death in pheochromocytoma 12 (PC12) cells. Reactive oxygen species (ROS), 2,2'-azinobis (3-ethylbebzothiazoloine-6-sulfonic acid) diammonium salt, and 1,1-diphenyl-2-picrrylhydrazyl radical scavenging activities, as well as the expression levels of proteins associated with oxidative damage and cell death were investigated. According to the results, C. lanceolate extract exhibited inhibitory activity against intracellular ROS generation and cell-damaging effects induced by hydroxyl radicals in a dose-dependent manner. Total phenolic and flavonoid contents were 22.3 mg·g^-1 gallic acid equivalent and 16.2 mg·g^-1 catechin equivalent, respectively. Additionally, a high-performance liquid chromatography (HPLC) assay based on the internal standard method used to detect phenolic compounds. The phenolic compounds identified in C. lanceolata extract contained (+)-catechin hydrate (5.0 ± 0.0 mg·g^-1), ferulic acid (1.6 ± 0.0 mg·g^-1), chlorogenic acid (1.5 ± 0.0 mg·g^-1), caffeic acid (1.2 ± 0.0 mg·g-1), naringin (0.9 ± 0.0 mg·g^-1), and p-coumaric acid (0.5 ± 0.0 mg·g^-1). C. lanceolata extract attenuated pro-apoptotic Bax expression levels and enhanced the expression levels of anti-apoptotic Bcl-2, caspase-3, and caspase-9 proteins. Therefore, C. lanceolata is a potential source of materials with neuroprotective properties against neurodegenerative disorders, such as Alzheimer's and Parkinson's diseases.

• Journal of Microbiology and Biotechnology
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• v.18 no.3
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• pp.523-531
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• 2008

Radiotherapy is currently applied in the treatment of human cancers. We studied whether genistein would enhance the radiosensitivity and explored its precise molecular mechanism in cervical cancer cells. After co-treatment with genistein and irradiation, the viability, cell cycle analysis, and apoptosis signaling cascades were elucidated in CaSki cells. The viability was decreased by co-treatment with genistein and irradiation compared with irradiation treatment alone. Treatment with only ${\gamma}$-irradiation led to cell cycle arrest at the $G_1$ phase. On the other hand, co-treatment with genistein and ${\gamma}$-irradiation caused a decrease in the $G_1$ phase and a concomitant increase up to 56% in the number of $G_2$ phase. In addition, co-treatment increased the expression of p53 and p21, and Cdc2-tyr-15-p, supporting the occurrence of $G_2/M$ arrest. In general, apoptosis signaling cascades were activated by the following events: release of cytochrome c, upregulation of Bax, down regulation of Bcl-2, and activation of caspase-3 and -8 in the treatment of genistein and irradiation. Apparently, co-treatment downregulated the transcripts of E6*I, E6*II, and E7. Genistein also stimulated irradiation-induced intracellular reactive oxygene, species (ROS) production, and co-treatment-induced apoptosis was inhibited by the antioxidant N-acetylcysteine, suggesting that apoptosis has occurred through the increase in ROS by genistein and ${\gamma}$-irradiation in cervical cancer cells. Gamma-irradiation increased cyclooxygenase-1 (COX-2) expression, whereas the combination with genistein and ${\gamma}$-irradiation almost completely prevented irradiation-induced COX-2 expression and $PGE_2$ production. Co-treatment with genistein and ${\gamma}$-irradiation inhibited proliferation through $G_2/M$ arrest and induced apoptosis via ROS modulation in the CaSki cancer cells.

• Journal of Embryo Transfer
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• v.31 no.3
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• pp.169-177
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• 2016

There is a growing interest in the application of primary hepatocytes for treatment of liver diseases in humans and for drug development. Several studies have focused on long-term survival and di-differentiation blocking of primary hepatocytes in an in vitro culture system. Therefore, the present study also aimed to optimize an in vitro culture system using primary rat hepatocytes. Primary rat hepatocytes from 6-week-old male Crl:CD rats were isolated using a modified two-step collagenase perfusion. Healthy $3.5{\times}10^6$ primary rat hepatocytes were seeded into a 2 dimensional (2D) culture in a 25T culture flask coated with collagen type I or into a 3D culture in a 125-ml spinner flask for 7 days. Production of plasma protein (ALB and TF), apoptosis (BAX and BCL2), and CYP (CYP3A1) related genes were compared between the 2D and 3D culture systems. The 3D culture system had an advantage over the 2D system because of the relatively high expression of ALB and low expression of BAX in the 3D system. However, the level of CYP3A1 did not improve in the 3D culture with and without the presence of a dexamethasone inducer. Therefore, 3D culture has an advantage for albumin production and primary rat hepatocyte survivability, but a low expression of CYP3A1 indicated that primary rat hepatocytes require a high-density culture for stress reduction by continuous flow.

• Proceedings of the Ginseng society Conference
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• 1998.06a
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• pp.231-243
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• 1998

In the present study, we provide evidence that ginsenoside $Rh_2$ (G-$Rh_2$) as well as staurosporine induces apoptosis of human hepatoma SK-HEP-1 cells by caspase 3-mediated processing of $p21^{WAFI/CIPI}$ in the early stage of apoptosls. Immunoblottings showed that G-$Rh_2$ as well as statrosporine induced the processing of caspase-3 to an active form, pl7. In stable Bcl-2 transfectants however, G-$Rh_2$ induced DNA fragmentation, while staurosporine did not. In the early stage of apoptosis, $p21^{WAFI/CIPI}$ was detected to undergo proteolytic processing specifically conducted by caspase-3. $p21^{WAFI/CIPI}$ translated in vitro was cleaved into a p14 fragment, when incubated with cell extracts obtained from either G-$Rh_2$- or staurosporine-treated cells. Cleavage was equally inhibited in both cases by adding Ac-DEVD-cho, a specific caspase-3 inhibitor, but not by Ac-YVkD-cho, a specific caspase-l inhibitor. Similarly, $p21^{WAFI/CIPI}$ was efficiently leaved by recombinant caspase-3 overexpressed in E. coli. Moreover, the endogenous $p21^{WAFI/CIPI}$ of untreated-cell extracts was also cleaved by recombinant caspase-3. Mutation analysis allowed identification of two caspase-3 cleavage sites, $DHVD^{112}$/L and $SMTD^{149}$/F, which are located within, or near the interaction domains for cyclins, Cdks, and PCNA. Taken together, these results show that G-$Rh_2$ as well as staurosporine increases caspase-3 activity, which in turn directly cleaves $p21^{WAFI/CIPI}$ resulting in elevation of Cdk kinase activity in the early stages of apoptosis. We propose that proteolytic cleavage of $p21^{WAFI/CIPI}$ is a functionally relevant event that allows unleashing the cyclin/Cdk activity from the inhibitor seen in the earlier stage of apoptosis, the event of which may be associated with the triggering mechanism for the execution of apoptosis.

• Proceedings of the Korean Vacuum Society Conference
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• 2010.02a
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• pp.406-406
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• 2010

일반적으로, 나노스케일의 MOS 소자에서는 게이트 절연체 두께가 감소함에 따라 tunneling effect의 증가로 인해 PID (plasma induced damage)로 인한 소자 특성 저하 현상을 감소하는 추세로 알려져 있다. 하지만 요즘 많이 사용되고 있는 high-k 게이트 절연체의 경우에는 오히려 더 많은 charge들이 trapping 되면서 PID가 오히려 더 심각해지는 현상이 나타나고 있다. 이러한 high-k 게이트 식각 시 현재는 주로 Hf-based wet etch나 dry etch가 사용되고 있지만 gate edge 영역에서 high-k 게이트 절연체의 undercut 현상이나 PID에 의한 소자특성 저하가 보고되고 있다. 본 연구에서는 이에 차세대 MOS 소자의 gate stack 구조중 issue화 되고 있는 metal gate 층과 gate dielectric 층의 식각공정에 각각 중성빔 식각과 중성빔 원자층 식각을 적용하여 전기적 손상 없이 원자레벨의 정확한 식각 조절을 해줄 수 있는 새로운 two step 식각 공정에 대한 연구를 진행하였다. 먼저 TiN metal gate 층의 식각을 위해 HBr과 $Cl_2$ 혼합가스를 사용한 중성빔 식각기술을 적용하여 100 eV 이하의 에너지 조건에서 하부층인 $HfO_2$와 거의 무한대의 식각 선택비를 얻었다. 하지만 100 eV 조건에서는 낮은 에너지에 의한 빔 스케터링으로 실제 패턴 식각시 etch foot이 발생되는 현상이 관찰되었으며, 이를 해결하기 위하여 먼저 높은 에너지로 식각을 진행하고 $HfO_2$와의 계면 근처에서 100 eV로 식각을 해주는 two step 방법을 사용하였다. 그 결과 anistropic 하고 하부층에 etch stop된 식각 형상을 관찰할 수 있었다. 다음으로 3.5nm의 매우 얇은 $HfO_2$ gate dielectric 층의 정확한 식각 깊이 조절을 위해 $BCl_3$와 Ar 가스를 이용한 중성빔 원자층 식각기술을 적용하여 $1.2\;{\AA}$/cycle의 단일막 식각 조건을 확립하고 약 30 cycle 공정시 3.5nm 두께의 $HfO_2$ 층이 완벽히 제거됨을 관찰할 수 있었다. 뿐만 아니라, vertical 한 식각 형상 및 향상된 표면 roughness를 transmission electron microscope(TEM)과 atomic force microscope (AFM)으로 관찰할 수 있었다. 이러한 중성빔 식각과 중성빔 원자층 식각기술이 결합된 새로운 gate recess 공정을 실제 MOSFET 소자에 적용하여 기존 식각 방법으로 제작된 소자 결과를 비교해 본 결과 gate leakage current가 약 one order 정도 개선되었음을 확인할 수 있었다.

• Radiation Oncology Journal
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• v.28 no.2
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• pp.91-98
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• 2010

Purpose: Troglitazone (TRO), a PPAR-$\gamma$ agonist, can reduce heat shock protein (HSP) 70 and increase the antioxidant enzymes, such as superoxide dismutase (SOD) and catalase, which might affect thermal sensitivity. Here, we investigated whether TRO modifies thermal sensitivity in uterine cervical cancer cells, which is most commonly treated by hyperthermia (HT). Materials and Methods: HeLa cells were treated with $5{\mu}M$ TRO for 24 hours before HT at $42^{\circ}C$ for 1 hour. Cell survival was analyzed by clonogenic assay. The expression of HSPs was analyzed by Western blot. SOD and catalase activity was measured and reactive oxygen species (ROS) was measured using 2',7'-dichlorofluorescin diacetate and dihydroethidium. Results: The decreased cell survival by HT was increased by preincubation with TRO before HT. Expression of HSP 70 was increased by HT however, it was not decreased by preincubation with TRO before HT. The decreased Bcl-2 expression by HT was increased by preincubation with TRO. SOD and catalase activity was increased by 1.2 and 1.3 times,respectively with TRO. Increased ROS by HT was decreased by preincubation with TRO. Conclusion: TRO decreases thermal sensitivity through increased SOD and catalase activity, as well as scavenging ROS in HeLa cells.

• Nutrition Research and Practice
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• v.10 no.1
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• pp.115-124
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• 2016

BACKGROUND/OBJECTIVES: This is the first study to identify common genetic factors associated with the basal metabolic rate (BMR) and body mass index (BMI) in obese Korean women including overweight. This will be a basic study for future research of obese gene-BMR interaction. SUBJECTS/METHODS: The experimental design was 2 by 2 with variables of BMR and BMI. A genome-wide association study (GWAS) of single nucleotide polymorphisms (SNPs) was conducted in the overweight and obesity (BMI > $23kg/m^2$) compared to the normality, and in women with low BMR (< 1426.3 kcal/day) compared to high BMR. A total of 140 SNPs reached formal genome-wide statistical significance in this study (P < $1{\times}10^{-4}$). Surveys to estimate energy intake using 24-h recall method for three days and questionnaires for family history, a medical examination, and physical activities were conducted. RESULTS: We found that two NRG3 gene SNPs in the 10q23.1 chromosomal region were highly associated with BMR (rs10786764; $P=8.0{\times}10^{-7}$, rs1040675; $2.3{\times}10^{-6}$) and BMI (rs10786764; $P=2.5{\times}10^{-5}$, rs10786764; $6.57{\times}10^{-5}$). The other genes related to BMI (HSD52, TMA16, MARCH1, NRG1, NRXN3, and STK4) yielded P < $10{\times}10^{-4}$. Five new loci associated with BMR and BMI, including NRG3, OR8U8, BCL2L2-PABPN1, PABPN1, and SLC22A17 were identified in obese Korean women (P < $1{\times}10^{-4}$). In the questionnaire investigation, significant differences were found in the number of starvation periods per week, family history of stomach cancer, coffee intake, and trial of weight control in each group. CONCLUSION: We discovered several common BMR- and BMI-related genes using GWAS. Although most of these newly established loci were not previously associated with obesity, they may provide new insights into body weight regulation. Our findings of five common genes associated with BMR and BMI in Koreans will serve as a reference for replication and validation of future studies on the metabolic rate.