• Journal of Microbiology and Biotechnology
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• 제27권1호
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• pp.197-205
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• 2017

Exposure of Jurkat T cell clone (J/Neo cells) to acacetin (5,7-dihydroxy-4'-methoxyflavone), which is present in barnyard millet (Echinochloa esculenta (A. Braun)) grains, caused cytotoxicity, enhancement of apoptotic $sub-G_1$ rate, Bak activation, loss of mitochondrial membrane potential (${\Delta}{\Psi}m$), activation of caspase-9 and caspase-3, degradation of poly(ADP-ribose) polymerase, and FITC-Annexin V-stainable phosphatidylserine exposure on the external surface of the cytoplasmic membrane without accompanying necrosis. These apoptotic responses were abrogated in Jurkat T cell clone (J/Bcl-xL) overexpressing Bcl-xL. Under the same conditions, cellular autophagic responses, including suppression of the Akt-mTOR pathway and p62/SQSTM1 down-regulation, were commonly detected in J/Neo and J/Bcl-xL cells; however, formation of acridine orange-stainable acidic vascular organelles, LC3-I/II conversion, and Beclin-1 phosphorylation (Ser-15) were detected only in J/Neo cells. Correspondingly, concomitant treatment with the autophagy inhibitor (3-methyladenine or LY294002) appeared to enhance acacetin-induced apoptotic responses, such as Bak activation, ${\Delta}{\Psi}m$ loss, activation of caspase-9 and caspase-3, and apoptotic $sub-G_1$ accumulation. This indicated that acacetin could induce apoptosis and cytoprotective autophagy in Jurkat T cells simultaneously. Together, these results demonstrate that acacetin induces not only apoptotic cell death via activation of Bak, loss of ${\Delta}{\Psi}m$, and activation of the mitochondrial caspase cascade, but also cytoprotective autophagy resulting from suppression of the Akt-mTOR pathway. Furthermore, pharmacologic inhibition of the autophagy pathway augments the activation of Bak and resultant mitochondrial damage-mediated apoptosis in Jurkat T cells.

• Journal of Korean Neurosurgical Society
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• 제63권5호
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• pp.566-578
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• 2020

Objective : Radiation is known to induce autophagy in malignant glioma cells whether it is cytocidal or cytoprotective. Dexamethasone is frequently used to reduce tumor-associated brain edema, especially during radiation therapy. The purpose of the study was to determine whether and how dexamethasone affects autophagy in irradiated malignant glioma cells and to identify possible intervening molecular pathways. Methods : We prepared p53 mutant U373 and LN229 glioma cell lines, which varied by phosphatase and tensin homolog (PTEN) mutational status and were used to make U373 stable transfected cells expressing GFP-LC3 protein. After performing cell survival assay after irradiation, the IC₅₀ radiation dose was determined. Dexamethasone dose (10 μM) was determined from the literature and added to the glioma cells 24 hours before the irradiation. The effect of adding dexamethasone was evaluated by cell survival assay or clonogenic assay and cell cycle analysis. Measurement of autophagy was visualized by western blot of LC3-I/LC3-II and quantified by the GFP-LC3 punctuated pattern under fluorescence microscopy and acridine orange staining for acidic vesicle organelles by flow cytometry. Results : Dexamethasone increased cell survival in both U373 and LN229 cells after irradiation. It interfered with autophagy after irradiation differently depending on the PTEN mutational status : the autophagy decreased in U373 (PTEN-mutated) cells but increased in LN229 (PTEN wild-type) cells. Inhibition of protein kinase B (AKT) phosphorylation after irradiation by LY294002 reversed the dexamethasone-induced decrease of autophagy and cell death in U373 cells but provoked no effect on both autophagy and cell survival in LN229 cells. After ATG5 knockdown, radiation-induced autophagy decreased and the effect of dexamethasone also diminished in both cell lines. The diminished autophagy resulted in a partial reversal of dexamethasone protection from cell death after irradiation in U373 cells; however, no significant change was observed in surviving fraction LN229 cells. Conclusion : Dexamethasone increased cell survival in p53 mutated malignant glioma cells and increased autophagy in PTEN-mutant malignant glioma cell but not in PTEN-wildtype cell. The difference of autophagy response could be mediated though the phosphatidylinositol 3-kinase/AKT/mammalian target of rapamycin signaling pathway.

• 대한환경공학회지
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• 제27권7호
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• pp.753-761
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• 2005

전처리 공정에 따른 투과 flux 변화를 살펴보면 $MIEX^{(R)}$+UF 공정의 경우 응집+UF 공정에 비하여 높은 유기물 제거율을 나타내었으나 투과 flux 감소는 크게 나타났다. 따라서, 막 오염을 자세하게 규명하기 위하여 sequential filtration 실험결과 막 오염물질은 고분자 유기물질로 나타났으며, DOC 0.5 mg/L 이하의 유기물질이 막 오염 유발물질로 작용하고 있었다. 10 kDa 이하의 저분자 유기물질들은 $MIEX^{(R)}$ 처리에 의해 제거가 용이하여 막 표면에서 유기물 부하를 감소시켜 이로 인해 응집+UF공정에 비하여 투과 flux 감소율이 낮게 나타났다. $MIEX^{(R)}$+UF공정은 입자상 물질의 존재 유무에 관계없이 flux 감소율은 거의 유사한 경향을 보였으나 응집+UF공정은 용존성 유기물질만이 존재하는 시수에 비하여 입자상 물질이 존재하는 경우에 투과 flux 감소가 더 낮게 나타났다.

• 대한화학회지
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• 제18권5호
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• pp.381-388
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• 1974

p-Phenylenediamine(PPD) 및 m-aminophenol(MAP)를 몰비를 달리하면서, -5∼0$^{\circ}$, $N_2$ 기류중에서 일정량의 포르말린(이때 포름알데히드의 량은 PPD와 MAP 전량에 대하여 2배몰에 해당하는 량으로 고정)과 반응시키면 약간의 발열과 함께 급격히 부가축합이 진행되어 난용성인 갈색 내지 등색의 중합체를 생성한다. PPD:MAP의 몰비가 1:3, 1:2, 1:1의 것들을 180∼220에서 몰비가 2:1, 3:1의 것들을 250∼270$^{\circ}$에서 각각 암색으로 색변화를 이르킬 따름이며, 이 중합체들은 모두 300$^{\circ}$까지 용융하지 않는다. 이들 중합체의 메틸렌블루에 대한 흡착력은 N2 기류중에서 중합시킨것을 묽은(7%) 수산화나트륨으로 처리하였을때 현저히 흡착력이 증가되어 PPD-MAP-F(F;포름알데히드)의 몰비가 1:3:8 일때 수지 1g당 80mg의 메틸렌블루를 흡착하며, 브롬페놀블루에대한 흡착력은 PPD-MAP-F의 몰비가 2:1:6일때가 최고이며, 수지 1g당 250mg의 브롬페놀블루를 흡착한다. TGA에 의한 내열성실험에서는 PPD-MAP-F의 몰비가 1:3:8인 경우 가장 좋은 결과를 나타냈으며, $N_2$ 기류중 매분 2$^{\circ}$로 가열하였을때 900$^{\circ}$에서 45%의 중량손실을 나타내었다.

• Journal of Ginseng Research
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• 제41권3호
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• pp.247-256
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• 2017

Background: KG-135, a standardized formulation enriched with Rk1, Rg3, and Rg5 ginsenosides, has been shown to inhibit various types of cancer cells; however, the underlying mechanisms are not fully understood. In this study, we explored its effects in A549 human lung cancer cells to investigate the induction of Forkhead Class box O3a (FOXO3a) and autophagy. Methods: Cell viability was determined by sulforhodamine B staining. Apoptosis and cell cycle distribution were analyzed using flow cytometry. The changes of protein levels were determined using Western blot analysis. Autophagy induction was monitored by the formation of acidic vesicular organelles stained with acridine orange. Results: KG-135 effectively arrested the cells in G1 phase with limited apoptosis. Accordingly, a decrease of cyclin-dependent kinase-4, cyclin-dependent kinase-6, cyclin D1, and phospho-retinoblastoma protein, and an increase of p27 and p18 proteins were observed. Intriguingly, KG-135 increased the tumor suppressor FOXO3a and induced the accumulation of autophagy hallmark LC3-II and acidic vesicular organelles without an increase of the upstream marker Beclin-1. Unconventionally, the autophagy adaptor protein p62 (sequestosome 1) was increased rather than decreased. Blockade of autophagy by hydroxychloroquine dramatically potentiated KG-135-induced FOXO3a and its downstream (FasL) ligand accompanied by the cleavage of caspase-8. Meanwhile, the decrease of Bcl-2 and survivin, as well as the cleavage of caspase-9, were also drastically enhanced, resulting in massive apoptosis. Conclusion: Besides arresting the cells in G1 phase, KG-135 increased FOXO3a and induced an unconventional autophagy in A549 cells. Both the KG-135-activated extrinsic FOXO3a/FasL/caspase-8 and intrinsic caspase-9 apoptotic pathways were potentiated by blockade of autophagy. Combination of KG-135 and autophagy inhibitor may be a novel strategy as an integrative treatment for cancers.

• 한국식품영양과학회지
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• 제45권4호
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• pp.577-584
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• 2016

다양한 환경에서 분리된 시가독소 생산성 대장균(Shiga toxin-producing E. coli, STEC, n=18)을 초산혼합용액(AAS;400 mM, pH 3.2, $30^{\circ}C$)에 노출한 후 산 저항성을 측정하였다. 또한, 선정된 4종류의 non-O157:H7 STEC균을 사과주스, 파인애플주스, 오렌지주스, 딸기주스(pH 3.8)에 정봉하여 $4^{\circ}C$와 $20^{\circ}C$에서 24시간 산 적응시킨 후 위합성용액(SGF, pH 1.5)에서 2시간 동안 생존능력을 평가하였다. Non-O157:H7 STEC를 AAS에 노출했을 때 O111 혈청형의 STEC는 평균 0.12 log CFU/mL 감소하여 다른 혈청형에 비하여 가장 강한 산 저항성을 나타냈고 O157:H7 STEC와 유의적 차이가 없었으며(P>0.05), O26 혈청형의 STEC는 가장 민감한 것으로 나타났다. 반면, AAS에 glutamic acid를 첨가하였을 경우 모든 STEC는 혈청형과 관계없이 초산에 매우 강한 저항성을 나타내었다(P>0.05). SGF에서 생존능력을 측정한 결과, 06E0218(O157:H7)은 다른 non-O157:H7 STEC 균들보다 생존능력이 낮았고 03-4669(O145:NM)가 가장 강한 생존능력을 나타내었다. 한편, 과일주스 중에서는 파인애플주스에 산 적응된 STEC가 SGF에 가장 강한 생존능력을 나타내었다. $4^{\circ}C$의 과일주스에 STEC를 산 적응시켰을 경우 $20^{\circ}C$보다 SGF에 대한 생존능력이 현저하게 높았다(P<0.05). 따라서 과일주스에 의한 non-O157:H7 STEC의 산 적응력 증가는 위장관 내 생존율 및 식중독 발생을 높일 수 있으므로 이에 대한 적절한 연구와 안전관리 옵션을 제공할 필요가 있을 것으로 판단된다.

• Restorative Dentistry and Endodontics
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• 제27권2호
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• pp.142-149
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• 2002

Low-viscosity composite resins may produce better sealed margins than stiffer compositions (KempScholte and Davidson, 1988: Crim, 1989). Plowable composites have been recommended for use in Class V cavities but it is also controversial because of its high rates of shrinkage. On the other hand, in the study comparing elastic moduli and leakage, the microfill had the least leakage (Rundle et at. 1997) Furthermore, in the 1996 survey of the Reality Editorial Team, microfills were the clear choice for abfraction lesions. The purpose of this study was to evaluate the microleakage of 6 compostite resins (2 hybrids, 2 microfills, and 2 flowable composites) with and without load cycling. Notch-shaped Class V cavities were prepared on buccal surface of 180 extracted human upper premolars on cementum margin. The teeth were randomly divided into non-load cycling group (group 1) and load cycling group (group 2) of 90 teeth each. The experimental teeth of each group were randomly divided into 6 subgroups of 15 samples. All preparations were etched, and Single bond was applied. Preparations were restored with the following materials (n=15) : hybrid composite resin [Z250(3M Dental Products Inc. St. Paul, USA), Denfil(Vericom, Ahnyang, Korea)], microfill [Heliomolar RO(Vivadent, Schaan, Liechtenstein), Micronew(Bisco Inc. Schaumburg, IL, USA)], and flowable composite［AeliteFlo(Bisco Inc. Schaumburg, IL, USA), Revolution(Kerr Corp. Orange, CA, USA)]. Teeth of group 2 were subjected to occlusal load (100N for 50,000 cycles) using chewing simulator(MTS 858 Mini Bionix II system, MTS Systems Corp. Minn. USA). All samples were coated with nail polish 1mm short of the restoration, placed in 2％ methylene blue for 24 hours, and sectioned with a diamond wheel. Enamel and dentin/cementum margins were analyzed for microleakage on a sclale of 0 (no leakage) to 3 (3/3 of wall). Results were statistically analyzed by Kruscal-Wallis One way analysis, Mann-Whitney U-test, and Student-Newmann-Keuls method. (p = 0.05) Results : 1. There was significantly less microleage in enamel margins than dentinal margins of all groups (p＜0.05) 2. There was no significant between six composite resin in enamel margin of group 1. 3. In dentin margin of group 1, flowable composite had more microleakage than others but not of significant differences. 4. there was no significant difference between six composite resin in enamel margin of group 2. 5. In dentin margin of group 2, the microleakage were R＞A =H=M＞D＞Z. But there was no significant differences. 6. In enamel margins, load cycling did not affect the marginal microleakage in significant degree. 7. In enamel margins, load cycling did affect the marginal microleakage only in Revolution. (p＜0.05).