• Journal of Microbiology and Biotechnology
• /
• v.27 no.1
• /
• pp.197-205
• /
• 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
• /
• v.63 no.5
• /
• pp.566-578
• /
• 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.

• Journal of Korean Society of Environmental Engineers
• /
• v.27 no.7
• /
• pp.753-761
• /
• 2005

In this study, pretreatment of organic matters with $MIEX^{(R)}$ was evaluated using bench-scale experimental procedures on four organic matters to determine its effect on subsequent UF membrane filtration. For comparison, coagulation process was also used as a pretreatment of UF membrane filtration. Moreover, the membrane fouling potential was identified using different fractions and molecular weights of organic matters. From the removal property of MW organic matters by coagulation process for the sample water NOM and AOM, the removal efficiency of high MW organic matters were much higher than those of low MW organic matters. It was shown that the removal efficiency of high MW organic matter more than 10 kDa was lower than that of low MW organic matter for $MIEX^{(R)}$ process. For the change of permeate flux by the pretreatment process, $MIEX^{(R)}$+UF process showed high removal efficiency of organic matter as compared with coagulation-UF processes, but high reduction rate of permeate flux was presented through the reduction of removal efficiency of high MW organic matter. From sequential filtration test results in order to examine the effect of MW of organic matter on membrane fouling, it was found that the membrane foulant was occurred by high MW organic matter, and the DOC of organic matter less than 0.5 mg/L was working as the membrane foulant. In the case of sample water composed of low MW organic matter less than 10 kDa, since the low MW organic matter less than 10 kDa has high removal efficiency by $MIEX^{(R)}$, low reduction rate of permeate flux is obtained as compared with coagulation-UF processes. In summary, it is required to conduct the research on physical/chemical characteristic of original water before pretreatment process of membrane process is selected, and a pertinent pre-treatment process should be employed based on the physical/chemical characteristic of original water.

• Journal of the Korean Chemical Society
• /
• v.18 no.5
• /
• pp.381-388
• /
• 1974

When mixture of p-phenylenediamine (PPD) and m-aminophenol (MAP) were reacted with formaldehyde (F) varying their amounts under $N_2$ stream at the temperature of -5∼0$^{\circ}$, addition condensation reaction occurred and brown colored resins(in some cases orange colored) were formed immediately. All resins thus formed were insoluble in most ordinary organic solvents and did not melt up to 300$^{\circ}$. When the resins were treated with dilute(7 %) aqueous sodium hydroxide solution, the adsorptivity of methylene blue on them showed marked improvement reaching as much as 80 mg of methylene blue on 1 g of the resin. On the other hand, in the case of bromophenol blue, its amount of adsorption appeared 250 mg per 1 g of the resin. The TGA under $N_2$ atmosphere indicated that the resin formed in molar ratio of 1 : 3 : 8 (PPD : MAP : F) showed the best heat-resistant property among others. About 40 % weight loss was observed for this resin at 900$^{\circ}$ with heating rate of 2$^{\circ}$ per minute.

• Journal of Ginseng Research
• /
• v.41 no.3
• /
• pp.247-256
• /
• 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.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.45 no.4
• /
• pp.577-584
• /
• 2016

The objectives of this study were I) to compare the acid resistance (AR) of seven non-O157 Shiga toxin-producing Escherichia coli (STEC) serogroups, including O26, O45, O103, O111, O121, O145, and O157:H7 STEC isolated from various sources, in 400 mM acetic acid solution (AAS) at pH 3.2 and $30^{\circ}C$ for 25 min with or without glutamic acid and II) to determine strain survival upon exposure to simulated gastric fluid (SGF, pH 1.5) at $37^{\circ}C$ for 2 h after acid adaptation in apple, pineapple, orange, and strawberry juices at pH 3.8, $4^{\circ}C$ and $20^{\circ}C$. Results show that the O111 serogroup strains had the strongest AR (0.12 log reduction CFU/mL) which was very similar to that of O157:H7 STEC (P>0.05), compared to other serogroups in AAS without glutamic acid, whereas O26 serogroup strains showed the most sensitive AR. However, there was no significant (P>0.05) difference of AR among seven serogroups in AAS with glutamic acid. In the SGF study, 05-6545 (O45:H2), 08023 (O121:H19), and 03-4669 (O145:NM) strains adapted in fruit juices at $4^{\circ}C$ and $20^{\circ}C$ displayed enhanced survival with exposure to SGF for 60 min compared to 06E0218 (O157:H7) strains (P<0.05). In addition, 4 STEC strains adapted in pineapple juice at $4^{\circ}C$ showed enhanced survival with exposure to SGF for 60 min compared to those strains acid-adapted in the other fruit juices. Generally, adaptation at $4^{\circ}C$ in fruit juices resulted in significantly enhanced survival levels compared to acid-adapted at $20^{\circ}C$ and non-adapted conditions. The AR caused by adaptation in fruit juices at low temperature may thus increase survival of non-O157 STEC strain in acidic environments such as the gastrointestinal tract. These results suggest that more careful strategies should be provided to protect against risk of foodborne illness by non-O157 STEC.

• Restorative Dentistry and Endodontics
• /
• v.27 no.2
• /
• pp.142-149
• /
• 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).