• Title/Summary/Keyword: LC model

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The Effect of Remifentanil Preconditioning on Injured Keratinocyte

  • Hong, Hun Pyo;Kim, Cheul Hong;Yoon, Ji Young;Kim, Yong Deok;Park, Bong Soo;Kim, Yong Ho;Yoo, Ji Uk
    • Journal of The Korean Dental Society of Anesthesiology
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    • v.14 no.3
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    • pp.157-165
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    • 2014
  • Background: Incisional site of surgical operation become transient ischemic state and then occur reoxygenation due to vasodilatation by inflammatory reaction, the productive reactive oxygen species (ROS) give rise to many physiologic results. Apoptosis have major role on elimination of inflammatory cell and formation of granulation tissue in normal wound healing process. Remifentanil can prevent the inflammatory response and can suppress inducible nitric oxide synthase expression in a septic mouse model. After cardiopulmonary bypass for coronary artery surgery, remifentanil can also inhibit the release of biomarkers of myocardial damage. Here we investigated whether remifentanil pretreatment has cellular protective effect against hypoxia-reoxygenation in HaCaT human keratinocytes, if so, the role of apoptosis and autophagy on this phenomenon. Methods: The HaCaT human keratinocytes were exposed to various concentrations of remifentanil (0.01, 0.05, 0.1, 0.5 and 1 ng/ml) for 2 h before hypoxia (RPC/HR group). These cells were cultured under 1% oxygen tension for 24h at $37^{\circ}C$. After hypoxia, to simulate reoxygenation and recovery, the cells were reoxygenated for 12 h at $37^{\circ}C$. 3-MA/RPC/HR group was treated 3-methyladenine (3-MA), autophagy inhibitor for 1h before remifentanil treatment. Cell viability was measured using a quantitative colorimetric assay with thiazolyl blue tetrazoliumbromide (MTT, amresco), showing the mitochondrial activity of living cells. To investigate whether the occurrence of autophagy and apoptosis, we used fluorescence microscopy and Western blot analysis. Results: The viability against hypoxia-reoxygenation injury in remifentanil preconditioning keratinocytes were increased, and these cells were showed stimulated expression of autophagy 3-MA suppressed the induction of autophagy effectively and the protective effects on apoptosis. Atg5, Beclin-1, LC3-II and p62 were elevated in RPC/HR group. But they were decreased when autophagy was suppressed by 3-MA. Conclusions: Remifentanil preconditioning showed the protective effect in human keratinocytes, and we concluded that autophagy may take the major role in the recovery of wound from hypoxia-reoxygenation injury. We suggest that further research is needed about the cell protective effects of autophagy.

Toxic Effects of Metal Plating Wastewater on Daphnia magna and Euglena agilis (Daphnia magna와 Euglena agilis를 이용한 도금폐수 독성평가)

  • Lee, Junga;Park, Da Kyung
    • Korean Journal of Environmental Biology
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    • v.34 no.2
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    • pp.116-123
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    • 2016
  • The ecotoxicity tests for metal plating wastewater were conducted using Daphnia magna (D. magna) and Euglena agilis (E. agilis). Evaluation for sources of toxicity was performed by 1) Correlation analysis between the concentration of individual metals in the metal plating wastewater and the toxic effects on D. magna, 2) Toxicant identification evaluation methods including graduated pH method, EDTA procedure and sodium thiosulfate procedure, 3) Comparison of toxic effect value ($EC_{50}$ or $LC_{50}$) of individual metal on D. magna and it's concentration in the metal plating wastewater. To evaluate the possibility of E. agilis, a Korean domestic organism, as a test model organism for metal plating waste water, E. agilis toxicity test was also assessed using on-line euglena ecotoxicity system (E-Tox system). Based on toxicant characterization test using D. magna, it was expected that SS, oxidants and heavy metals are responsible for toxicity of metal plating waste water. Especially Cu, Hg, and Ag were the major cationic metals that caused toxicity. E. agilis is less sensitive than D. magna based on the $EC_{50}$ value however it shows prompt response to toxic test substances. E. agilis shows even a significant effect on the cell swimming velocity within 2 min to toxic metal plating wastewater. Our study demonstrates that E. agilis test can be a putative ecotoxicity test for assessing the quality of metal plating waste water.

A STUDY ON THE COLOR STABILITY OF RESIN MODIFIED GLASS IONOMERS (레진 강화형 글라스아이오노머의 색 안정성에 관한 연구)

  • Koo, Dae-Hoi;Lee, Yong-Keun;Son, Ho-Hyun;Im, Mi-Kyung
    • Restorative Dentistry and Endodontics
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    • v.22 no.1
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    • pp.358-373
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    • 1997
  • Resin modified glass ionomers were introduced in 1988 to overcome the problems of moisture sensitivity and low early mechanical strength of conventional glass ionomers and to maintain their clinical advantages. The purpose of this study was to evaluate the color stability of four resin modified glass ionomers(Fuji II LC, Vitremer, Dyract and VariGlass), one resin composite material(Z-100), and one conventional glass ionomer(GC Fuji II) under several conditions. These conditions were as follows: 1) before curing, 2) after curing, 3) after polishing, 4) after 500 thermocycling, 5) after 1,000 thermocycling, 6) after 1,500 thermocycling and 7) after 2,000 thermocycling. Three specimens of each material/shade combination were made. Materials were condensed into metal mold with a diameter of 10 mm and a thickness of 2.0 mm, and were pressed between glass plates. The material was polymerized using a light polymerizing unit(Visilux II, 3M, USA). After removal of excess, the surface was polished sequentially on wet sandpapers. A reflection spectrophotometer(Model TC-6FX, Tokyo Denshoku Co., Japan) was used to determine CIELAB coordinates($L^*,a^*$ and $b^*$) of each specimen. CIE standard illumination C was used as the light source. The results were as follows : 1. In comparing different shades of same material, CIELAB color difference(${\Delta}E^*$) value was not significantly different from each other(p>0.05). 2. CIELAB color difference(${\Delta}E^*$) values between after-curing and after-polishing were ranged from 5.53 to 27.08. These values were higher than those of other condition combinations. 3. CIELAB color difference(${\Delta}E^*$) values between before-thermocycling and after-thermocycling were ranged from 1.40 to 7.81. Despite the number of thermocycling increased, CIELAB color difference(${\Delta}E^*$) value was low. 4. The color stability of resin modified glass ionomers was more stable than that of conventional glass ionomers but less stable than that of Z100.

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Protective Effect of Propofol against Hypoxia-reoxygenation Injury in HaCaT Human Keratinocytes

  • Kim, Yong-Ho;Kang, Jin-Mo;Kim, In-Ryoung;Lee, Bo-Young;Yoon, Ji-Young;Kim, Cheul-Hong;Park, Bong-Soo
    • International Journal of Oral Biology
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    • v.39 no.2
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    • pp.97-105
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    • 2014
  • The aim of this study was to determine the beneficial effect of propofol on human keratinocytes that have undergone hypoxia reoxygenation (H/R) injury and to investigate whether autophagy is associated with the protective mechanism. Thus, we evaluated how propofol influences the intracellular autophagy and apoptosis during the H/R process in the HaCaT cells. The cultured human keratinocyte cells were exposed to 24 h of hypoxia (5% $CO_2$, 1% $O_2$, 94% $N_2$) followed by 12 h of reoxygenation (5% $CO_2$, 21% $O_2$, 74% $N_2$). The experiment was divided into 4 groups: (1) Control=Normoxia ; (2) H/R=Hypoxia Reoxygenation ; (3) PPC+H/R=Propofol Preconditioning+Hypoxia Reoxygenation; (4) 3-MA+PPC+ H/R=3-MA-Methyladenine+Propofol Preconditioning+ Hypoxia Reoxygenation. In addition, Western blot analysis was performed to identify the expression of apoptotic pathway parameters, including Bcl-2, Bax, and caspase 3 involved in mitochondrial-dependent pathway. Autophagy was determined by fluorescence microscopy, MDC staining, AO staining, and western blot. The H/R produced dramatic injuries in keratinocyte cells. In our study, the viability of Propofol in H/R induced HaCaT cells was first studied by MTT assay. The treatment with 25, 50, and $100{\mu}M$ Propofol in H/R induced HaCaT cells enhanced cell viability in a dose-dependent manner and $100{\mu}M$ was the most effective dose. The Atg5, Becline-1, LC3-II, and p62 were elevated in PPC group cells, but H/R-induced group showed significant reduction in HaCaT cells. The Atg5 were increased when autophagy was induced by Propofol, and they were decreased when autophagy was suppressed by 3-MA. These data provided evidence that propofol preconditioning induced autophagy and reduced apoptotic cell death in an H/R model of HaCaT cells, which was in agreement with autophagy playing a very important role in cell protection.

Remifentanil Protects Human Keratinocyte Through Autophagic Expression

  • Kim, Eok Nyun;Park, Chang Hoon;Woo, Mi Na;Yoon, Ji Young;Park, Bong Soo;Kim, Yong Ho;Kim, Cheul Hong
    • Journal of The Korean Dental Society of Anesthesiology
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    • v.14 no.2
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    • pp.101-106
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    • 2014
  • Background: Remifentanil, an ultra-short-acting mu-opioid receptor agonist, is unique from other opioids because of its esterase-based metabolism, minimal accumulation, and very rapid onset and offset of clinical action. Remifentanil can prevent the inflammatory response and can suppress inducible nitric oxide synthase expression in a septic mouse model. However, the effects of remifentanil on human keratinocyte and autophagy have yet to be fully elucidated during hypoxia-reoxygenation. Here we investigated whether remifentanil confers protective effect against hypoxia-reoxygenation in human keratinocyte and, if so, whether autophagy mediates this effect. Methods: The human keratinocytes were cultured under 1% oxygen tension. The cells were gassed with 94% $N_2$, and 5% $CO_2$ and incubated for 24 h at $37^{\circ}C$. To determine whether the administration of affects human keratinocytes hypoxia-reoxygenation injury, cells were then exposed to various concentrations of remifentanil (0.01, 0.1, 0.5 and 1 ng/ml) for 2 h. After remifentanil treatment, to simulate reoxygenation and recovery, the cells were reoxygenated for 12 h at $37^{\circ}C$. Control group did not receive remifentanil treatment. Normoxia group did not receive hypoxia and remifentanil treatment for 36 h. 3-MA group was treated 3-methyladenine (3-MA) for 1h before remifentanil treatment. Cell viability was measured using a quantitative colorimetric assay with MTT, showing the mitochondrial activity of living cells. Cells were stained with fluorescence and analyzed with Western blot analysis to find out any relations with activation of autophagy. Results: Prominent accumulation of autophagic specific staining MDC was observed around the nuclei in RPT group HaCaT cells. Similarly, AO staining, red fluorescent spots appeared in RPT group HaCaT cells, while the Normoxia, control and 3-MA groups showed mainly green cytoplasmic fluorescence. We here examined activation of autophagy related protein under H/R-induced cells by Western blotting analysis. Atg5, Beclin-1, LC3-II (microtubule-associated protein 1 light chain 3 form II) and p62 was elevated in RPT group cells. But they were decreased when autophagy was suppressed by 3-MA (Fig. 5). Conclusions: Although the findings of this study are limited to an in vitro interpretation, we suggest that remifentanil may have a beneficial effect in the recovery of wound from hypoxia-reoxygenation injury.

Cardioprotective effect of ginsenoside Rb1 via regulating metabolomics profiling and AMP-activated protein kinase-dependent mitophagy

  • Hu, Jingui;Zhang, Ling;Fu, Fei;Lai, Qiong;Zhang, Lu;Liu, Tao;Yu, Boyang;Kou, Junping;Li, Fang
    • Journal of Ginseng Research
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    • v.46 no.2
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    • pp.255-265
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    • 2022
  • Background: Ginsenoside Rb1, a bioactive component isolated from the Panax ginseng, acts as a remedy to prevent myocardial injury. However, it is obscure whether the cardioprotective functions of Rb1 are related to the regulation of endogenous metabolites, and its potential molecular mechanism still needs further clarification, especially from a comprehensive metabolomics profiling perspective. Methods: The mice model of acute myocardial ischemia (AMI) and oxygen glucose deprivation (OGD)-induced cardiomyocytes injury were applied to explore the protective effect and mechanism of Rb1. Meanwhile, the comprehensive metabolomics profiling was conducted by high-performance liquid chromatography and quadrupole time-of-flight mass spectrometry (HPLC-Q/TOF-MS) and a tandem liquid chromatography and mass spectrometry (LC-MS). Results: Rb1 treatment profoundly reduced the infarct size and attenuated myocardial injury. The metabolic network map of 65 differential endogenous metabolites was constructed and provided a new inspiration for the treatment of AMI by Rb1, which was mainly associated with mitophagy. In vivo and in vitro experiments, Rb1 was found to improve mitochondrial morphology, mitochondrial function and promote mitophagy. Interestingly, the mitophagy inhibitor partly attenuated the cardioprotective effect of Rb1. Additionally, Rb1 markedly facilitated the phosphorylation of AMP-activated protein kinase α (AMPKα), and AMPK inhibition partially weakened the role of Rb1 in promoting mitophagy. Conclusions: Ginsenoside Rb1 protects acute myocardial ischemia injury through promoting mitophagy via AMPKα phosphorylation, which might lay the foundation for the further application of Rb1 in cardiovascular diseases.

Antioxidant Activity of Novel Casein-Derived Peptides with Microbial Proteases as Characterized via Keap1-Nrf2 Pathway in HepG2 Cells

  • Zhao, Xiao;Cui, Ya-Juan;Bai, Sha-Sha;Yang, Zhi-Jie;Cai, Miao;Megrous, Sarah;Aziz, Tariq;Sarwar, Abid;Li, Dong;Yang, Zhen-Nai
    • Journal of Microbiology and Biotechnology
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    • v.31 no.8
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    • pp.1163-1174
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    • 2021
  • Casein-derived antioxidant peptides by using microbial proteases have gained increasing attention. Combination of two microbial proteases, Protin SD-NY10 and Protease A "Amano" 2SD, was employed to hydrolyze casein to obtain potential antioxidant peptides that were identified by LC-MS/MS, chemically synthesized and characterized in a oxidatively damaged HepG2 cell model. Four peptides, YQLD, FSDIPNPIGSEN, FSDIPNPIGSE, YFYP were found to possess high 1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging ability. Evaluation with HepG2 cells showed that the 4 peptides at low concentrations (< 1.0 mg/ml) protected the cells against oxidative damage. The 4 peptides exhibited different levels of antioxidant activity by stimulating mRNA and protein expression of the antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px), as well as nuclear factor erythroid-2-related factor 2 (Nrf2), but decreasing the mRNA expression of Kelch-like ECH-associated protein 1 (Keap1). Furthermore, these peptides decreased production of reactive oxygen species (ROS) and malondialdehyde (MDA), but increased glutathione (GSH) production in HepG2 cells. Therefore, the 4 casein-derived peptides obtained by using microbial proteases exhibited different antioxidant activity by activating the Keap1-Nrf2 signaling pathway, and they could serve as potential antioxidant agents in functional foods or pharmaceutic preparation.

A Qualitative Study of Physicians' Perspectives on Non-Cancer Hospice-Palliative Care in Korea: Focus on AIDS, COPD and Liver Cirrhosis (국내의 비암성 질환의 호스피스 완화의료 적용에 대한 전문가의 인식에 관한 질적 연구: 후천성 면역결핍 증후군, 만성 폐쇄성 폐질환, 간경화를 중심으로)

  • Shin, Jinyoung;Yoon, Seok-Joon;Kim, Sun-Hyun;Lee, Eon Sook;Koh, Su-Jin;Park, Jeanno
    • Journal of Hospice and Palliative Care
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    • v.20 no.3
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    • pp.177-187
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    • 2017
  • Purpose: From August 2017, hospice-palliative care (HPC) will be provided to patients with acquired immunodeficiency syndrome (AIDS), chronic obstructive pulmonary disease (COPD), and liver cirrhosis in Korea. To contribute to building a non-cancer (NC) hospice-palliative care model, NC specialists were interviewed regarding the goals, details, and provision methods of the model. Methods: Four physicians specializing in HPC of cancer patients formulated a semi-structured interview with questions extracted from literature review of 85 articles on NC HPC. Eleven NC disease specialists were interviewed, and their answers were analyzed according to the qualitative content analysis process. Results: The interviewees said as follows: It is difficult to define end-stage NC patients. HPC for cancer patients and that for NC patients share similar goals and content. However, emphasis should be placed on alleviating other physical symptoms and emotional care rather than pain control. Timing of the care provision should be when patients are diagnosed as "end stage". Special issues should be considered for each NC disease (e.g., use of anti-retroviral drugs for AIDS patients, oxygen supply for COPD patients suffering from dyspnea, liver transplantation for patients with liver cirrhosis) and education should be provided to healthcare professionals. NC patients tend to negatively perceive HPC, and the government's financial assistance is insufficient. Conclusion: It is necessary to define end-stage NC patients through in-depth discussion to minimize issues that will likely accompany the expansion of care recipients. This requires cooperation between medical staff caring for NC patients and HPC givers for cancer patients.

Hypoglycemic Effect of Culture Broth of Bacillus subtilis S10 Producing 1-Deoxynojirimycin (1-Deoxynojirimycin을 생산하는 Bacillus subtilis S10 배양액의 혈당강하 효과)

  • Cho, Yong-Seok;Park, Young-Shik;Lee, Jae-Yeon;Kang, Kyung-Don;Hwang, Kyo-Yeol;Seong, Su-Il
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.11
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    • pp.1401-1407
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    • 2008
  • 1-Deoxynojirimycin (DNJ) is a strong $\alpha$-glucosidase inhibitor which inhibits hyperglycemia in animals. To select the Bacillus strains highly producing DNJ, 4,000 strains were isolated from soil and grain samples. By the inhibitory activity against $\alpha$-glucosidase, nine Bacillus strains were selected and then identified by 16S rDNA sequencing. B. subtilis S10 was finally selected as the best strain for the production of DNJ. Various carbon sources and nitrogen sources in culture medium were evaluated for the highest production of DNJ. As the results, the optimized concentration of carbon source and nitrogen source was 1.0% galactose and 1.6% polypeptone and the concentration of DNJ produced was 0.75 g/L. The effect of culture supernatant of B. subtilis S10 on lowering blood glucose level was investigated in streptozotocin (STZ)-induced diabetic mice model. Mice were randomly assigned to control group (saline) and three test groups such as acarbose group, silkworm powder group and B. subtilis S10 group. After eight-week oral feeding, blood glucose levels of the B. subtilis S10 and silkworm powder groups were respectively $209.1{\pm}19.6\;mg/dL$ (59.1%) and $208.6{\pm}39.8\;mg/dL$ (59.0%) lower than $510{\pm}10\;mg/dL$ of the control group. These results indicated that the culture supernatant of B. subtilis S10 was able to reduce the blood glucose level in STZ-induced diabetic mice.