• Title/Summary/Keyword: Biological pretreatment

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Suppressive effects of ethanol extract of Aralia elata on UVB-induced oxidative stress in human keratinocytes (자외선 B를 조사한 인간유래각질세포에서 두릅순 에탄올추출물의 산화적 스트레스 억제효과)

  • Kwak, Chung Shil;Yang, Jiwon
    • Journal of Nutrition and Health
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    • v.49 no.3
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    • pp.135-143
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    • 2016
  • Purpose: Ultraviolet (UV)-induced oxidative stress contributes to several adverse biological effects on skin. Many phenolic phytochemicals have been shown to have antioxidant properties and protect skin cells from UV-induced oxidative damage. In this study, we investigated whether or not Aralia elata (AE) has a protective effect against UVB-induced reactive oxygen species (ROS), ultimately leading to photoaging. Methods: Phenolic content of dried AE and antioxidant properties of AE extract in 70% ethanol weredetermined by measuring DPPH and ABTS radical scavenging activities and ferric reducing antioxidant power (FRAP). The effect of AE extract on cellular ROS generation and expression levels of oxidative stress-response proteins such as superoxide dismutase (SOD)-1, catalase, nuclear factor-erythroid 2-related factor (Nrf)-2, and heme oxygenase (HO)-1 in UVB-irradiated ($75mJ/cm^2$) human keratinocytes (HaCaT) were further determined by 2'-7'-dichlorofluoresceine diacetate assay and Western blotting, respectively. Results: The total phenolic and flavonoid contents of dried AE were 20.15 mg tannic acid/g and 18.75 mg rutin/g, respectively. The $IC_{50}$ of AE extract against DPPH radical was $98.5{\mu}g/mL$, and ABTS radical scavenging activity and FRAP upon treatment with $1,000{\mu}g/mL$ of AE extract were $41.8{\mu}g\;ascorbic\;acid\;(AA)\;eq./mL$ and $29.7{\mu}g\;AA\;eq./mL$,m respectively. Pretreatment with AE extract significantly reduced (p < 0.05) ROS generation compared to that in UVB-irradiated control HaCaT cells. Pretreatment with AE extract reversed reduction of Nrf-2 and SOD-1 protein expression and induction of HO-1 protein expression caused by UVB exposure in HaCaT cells, whereas it did not affect catalase expression. Conclusion: AE extract in 70% ethanol demonstrated a protective effect against UVB-induced oxidative stress and decreased expression of Nrf-2 and SOD-1 in human keratinocytes. These findings suggest that AE ethanol extract might have potential as a natural resource for a skin anti-photoaging product in the food and cosmetic industry.

Adhesion-induced generation of oxygen free radical from human alveolar macrophages and its mechanisms (폐포대식세포의 부착에 의한 산소유리기 분비능 활성화 및 그 기전)

  • Chung, Man-Pyo;Yoo, Chul-Gyu;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo;Han, Yong-Chol
    • Tuberculosis and Respiratory Diseases
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    • v.43 no.2
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    • pp.210-220
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    • 1996
  • Background : Neutrophils or monocytes separated in vitro by the adherence to plastic surface are known to be activated by surface adherence itself and subsequent experimental data might be altered by surface adherence. In the process of surface adherence, adhesion molecules have a clear role in intracellular signal pathway of cellular activation. Human alveolar macrophages(HAM) are frequently purified by the adherence procedure after bronchoalveolar lavage. But the experimental data of many reports about alveolar macrophages have ignored the possibility of adhesion-induced cellular activation. Method : Bronchoalveolar lavage was performed in the person whose lung of either side was confirmed to be normal by chest CT. With the measurement of hydrogen peroxide release from adherent HAM to plastic surface and non-adherent HAM with or without additional stimulation of phorbol myristate acetate(PMA) or N-formyl-methionyl-leucyl-phenylalanine (fMLP), we observed the effect of the adherence to plastic surface. We also evaluated the effect of various biological surfaces on adhesion-induced activation of HAM. Then, to define the intracellular pathway of signal transduction, pretreatment with cycloheximide, pertussis toxin and anti-CD11/CD18 monoclonal antibody was done and we measured hydrogen peroxide in the culture supernatant of HAM. Results : 1) The adherence itself to plastic surface directly stimulated hydrogen peroxide release from human alveolar macrophages and chemical stimuli such as phorbol myristate acetate(PMA) or N-formyl-methionyl-leucyl-phenylalanine(fMLP) colud not increase hydrogen peroxide release in these adherent macrophages which is already activated. 2) PMA activated human alveolar macrophages irrespective of the state of adhesion. However, fMLP stimulated the release of hydrogen peroxide from the adherent macrophages, but not from the non-adherent macrophages. 3) HAM adherent to A549 cell(type II alveolar epithelium-like human cell line) monolayer released more hydrogen peroxide in response to both PMA and fMLP. This adherence-dependent effect of fMLP was blocked by pretreatment of macrophages with cycloheximide, pertussis toxin and anti-CD18 monoclonal antibody, Conclusion : These results suggest that the stimulatory effect of PMA and fMLP can not be found in adherent macrophage because of the activation of human alveolar macrophage by the adherence to plastic surface and the cells adhered to biologic surface such as alveolar epithelial cells are appropriately responsive to these stimuli. It is also likely that the effect of fMLP on the adherent macrophage requires new protein synthesis via G protein pathway and is dependent on the adhesion between alveolar macrophages and alveolar epithelial cells by virtue of CD11/CD18 adhesion molecules.

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A Protocol of Ludox Treatment for Physiological and Molecular Biological Research of Freshwater Cyanobacteria (퇴적층 남조류 휴면세포의 생리적-분자생물학적 연구를 위한 Ludox 처리법)

  • Keonhee Kim;Kyeong-eun Yoo;Hye-in Ho;Chaehong Park;Hyunjin Kim;Soon-Jin Hwang
    • Korean Journal of Ecology and Environment
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    • v.56 no.1
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    • pp.94-103
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    • 2023
  • Cyanobacterial resting cells, such as akinetes, are important seed cells for cyanobacteria's early development and bloom. Due to their importance, various methods have been attempted to isolate resting cells present in the sediment. Ludox is a solution mainly used for cell separation in marine sediments, but finding an accurate method for use in freshwater is difficult. This study compared the two most commonly used Ludox methods (direct sediment treatment and sediment distilled water suspension treatment). Furthermore, we proposed a highly efficient method for isolating cyanobacterial resting cells and eDNA amplification from freshwater sediments. Most of the resting cells found in the sediment were akinete to the Nostocale and were similar to those of Dolichospermum, Cylindrospermum, and Aphanizomenon. Twenty times more akinetes were found in the conical tube column using the sediment that had no treatment than in the sample treated by suspending the sediment in distilled water. Akinete separated through Ludox were mainly spread over the upper and lower layers in the column rather than concentrated at a specific depth in the column layer. The mibC, Geo, and 16S rDNA genes were successfully amplified using the sediment directly in the sample. However, the amplification products of all genes were not found in the sample in which the sediment was suspended in distilled water. Therefore, 5 g to 10 g of sediment is used without pretreatment when isolating cyanobacterial resting cells from freshwater sediment. Cell isolation and gene amplification efficiency are high when four times the volume of Ludox is added. The Ludox treatment method presented in this study isolates cyanobacterial resting cells in freshwater sediment, and the same efficiency may not appear in other biotas. Therefore, to apply Ludox to the separation of other biotas, it is necessary to conduct a pre-experiment to determine the sediment pretreatment method and the water layer where the target organism exists.

Effects of Platycodon grandiflorum on the Induction of Autophagy and Apoptosis in HCT-116 Human Colon Cancer Cells (길경 추출물에 의한 HCT-116 대장암 세포주에서의 autophagy와 apoptosis 유발 효과)

  • Hong, Su Hyun;Park, Cheol;Han, Min Ho;Kim, Hong Jae;Lee, Moon Hee;Choi, Yung Hyun
    • Journal of Life Science
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    • v.24 no.11
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    • pp.1244-1251
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    • 2014
  • Platycodon grandiflorum (PG) has been known to possess many biological effects, including anti-inflammatory and anti-allergy activity and anti-obesity and hyperlipidemia effects. However, little research has been conducted regarding its anticancer effects, with the exception of its ability to stimulate apoptosis in skin cells. There has also been no study regarding PG-induced autophagy. The modulation of autophagy is recognized as one of the hallmarks of cancer cells. Depending on the type of cancer and the context, autophagy can suppress or help cancer cells to overcome metabolic stress and the cytotoxicity of chemotherapy. Therefore, the present study was designed to investigate whether or not extracts from PG-induced cell death were connected with autophagy and apoptosis in HCT-116 human colon cancer cells. PG stimulation decreased cell proliferation in a dose- and time-dependent manner and induced apoptosis, which was partially dependent on the activation of caspases. PG treatment also resulted in the formation of autophagic vacuoles simultaneously with regulation of autophagy-related genes. Interestingly, a PG-mediated apoptotic effect was further triggered by pretreatment with the autophagy inhibitors 3-methyladenin and bafilomycin A1. However, cell viability recovered quite well with bafilomycin A1 treatment. These findings show that PG treatment promotes both autophagy and apoptosis and that PG-induced autophagic response might play a role in the autophagic cell death of HCT-116 cells.

Degradation of Fat, Oil, and Grease (FOGs) by Lipase-Producing Bacterium Pseudomonas sp. Strain D2D3

  • Shon, Ho-Kyong;Tian, Dan;Kwon, Dae-Young;Jin, Chang-Suk;Lee, Tae-Jong;Chung, Wook-Jin
    • Journal of Microbiology and Biotechnology
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    • v.12 no.4
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    • pp.583-591
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    • 2002
  • Biodegradation of fat, oil, and grease (FOGs) plays an Important role in wastewater management and water pollution control. However, many industrial food-processing and food restaurants generate FOG-containing waste waters for which there Is no acceptable technology for their pretreatment. To solve these problems, this study evaluated the feasibility of effective FOG-degrading microorganisms on the biodegradation of olive oil and FOG-containing wastewater. Twenty-two strains capable of degrading FOGs were isolated from five FOG-contaminated sites for the evaluation of their FOG degradation capabilities. Among twenty-two strains tested, the lipase-producing Pseudomonas sp. strain D2D3 was selected for actual FOG wastewater treatment. Its biodegradability was performed at 3$0^{\circ}C$ and pH 8. The extent of FOG removal efficiency was varied for each FOG tested, being the highest for olive oil and animal fat (94.5% and 94.4%), and the lowest for safflower oil (62%). The addition of organic nitrogen sources such as yeast extract, soytone, and peptone enhanced the removal efficiency of FOGs, but the addition of the inorganic nitrogen nutrients such as $NH_4$Cl and $(NH_4)_2SO_4$ did not increase. The $KH_2PO_4$ sources in 0.25% to 0.5% concentrations showed more than 90% degradability. As a result, the main pathway for the oxidation of fatty acids results in the removal of two carbon atoms as acetyl-CoA with each reaction sequence: $\beta$-oxidation. Its lipase activity showed 38.5 U/g DCW using the optimal media after 9 h. Real wastewater and FOGs were used for determining the removal efficiency by using Pseudomonas sp. strain D2D3 bioadditive. The degradation by Pseudomonas sp. strain D2D3 was 41% higher than that of the naturally occurring bacteria. This result indicated that the use of isolated Pseudomonas sp. strain D2D3 in a bioaugmentating grease trap or other processes might possibly be sufficient to acclimate biological processes for degrading FOGs.

The Protective Activity of Soeumin Bojungykgi-tang Water Extract Against Oxidative Stress-induced Hepato-Toxicity (산화적 스트레스로 유도된 간손상에 대한 소음인보중익기탕 열수추출물의 간세포보호효과)

  • Son, Jin Won;Jung, Ji Yun;Kim, Kwang-Youn;Hwangbo, Min;Park, Chung A;Cho, IL Je;Back, Young Doo;Jung, Tae Young;Kim, Sang Chan;Jee, Seon Young
    • Herbal Formula Science
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    • v.25 no.4
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    • pp.509-526
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    • 2017
  • Background and objectives : Soeumin Bojungykgi-tang (seBYTE) has been used to supplement qi in Korean medicine. It has been demonstrated to possess various biological functions such as anti-cancer, anti-aging and anti-inflammatory effects. The present study evaluated the protective roles of seBYTE in hepatotoxic in vitro and in vivo model. Methods : To investigate cytoprotective effect of seBYTE, HepG2 cells were pretreated with seBYTE and then subsequently exposed to $10{\mu}m$ AA for 12 h, followed by $5{\mu}m$ iron. Cell viability was examined by MTT assay, and expression of apoptosis-related proteins was evaluated by immunoblot analysis. For responsible molecular mechanisms, ROS production, GSH contents, and mitochondrial membrane potential were measured. In addition, hepatoprotective effect of seBYTE in vivo was assessed in $CCl_4$-induced animal model. Results : seBYTE prevented AA + iron-induced cytotoxicity in concentration dependent manner. In addition, ROS production, GSH depletion, and mitochondrial dysfunction induced by AA + iron were significantly reduced by seBYTE pretreatment. Furthermore, seBYTE recovered expression of the pro-apoptotic proteins such as PARP and pro-caspase-3. In animal experiment, plasma ALT and AST levels were significantly elevated in $CCl_4$ treatment, but seBYTE significantly decreased the ALT and AST levels. Moreover, seBYTE alleviated the numbers of histological activity index, percentages of degenerative regions, degenerated hepatocytes, infiltrated inflammatory cells, nitrotyrosine- and 4-hydroxynonenal-positive cells in liver. Conclusions : These results showed that hepatoprotective effect of seBYTE against on $CCl_4$-induced hepatic damages is partly due to antioxidative and anti-apoptotic process.

A Practical Study on the Solid-Liquid Separation of the Swine Wastewater from Slurry Feedlot (슬러리 양돈분뇨의 최적 고액분리 방안 연구)

  • Park, Seung-Kyun;Choi, Jae-Gil;Chung, Yoon-Jin
    • Journal of the Korea Organic Resources Recycling Association
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    • v.8 no.2
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    • pp.60-70
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    • 2000
  • The swine wastewater from slurry feedlot has been a social problem in Korea since the proper treatment is very difficult. Therefore, a practical study on the Solid-Liquid separation of swine wastewater from slurry feedlot was carried out as a pan of pretreatment for the successful biological treatment. The appropriate type of coagulant and optimum dosage were proposed for the most efficient Solid-Liquid separation and the best Solid-Liquid separation methods for different size of feedlot were determined through the tests with field-scaled Solid-Liquid separation equipment. The appropriate coagulant for the conditioning of dewatering property was E-851, which is a cationic polyelectrolyte made of polyacrylamide, and the optimum dosage was 0.24~0.6% of unit solids weight. Mesh Screen, Drum Screen, Cyclone Drum Filter, Screw Press, High-speed Screw Decanter, Low-speed Screw Decanter, and Dissolved Air Flotation Process had been investigated in this study. According to the results, the Screw Press was the best dewatering equipment for the small & medium size for feedlot and low-speed Screw Decanter was the best for the large size feedlot & public owned treatment facilities for the primary Solid-Liquid separation, and the most suitable secondary treatment process was DAF. On the other hand, reductions for the requirement of bulking agent and organic loading by Solid-Liquid separation process were 94.8% and 84.7%, respectively Therefore, the Solid-Liquid separation process must be required for the successful treatment of swine wastewater from slurry feedlot.

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Effect of Ozonation on Removal of Dissolved Organic Matter by Granular Activated Carbon Process (오존공정이 입상활성탄공정에서 용존유기물질의 제거에 미치는 영향)

  • Ahn, Hyo-Won;Chae, Seon-Ha;Wang, Chang-Keun;Lim, Jae-Lim
    • Journal of Korean Society of Environmental Engineers
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    • v.30 no.6
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    • pp.601-608
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    • 2008
  • The objective of this study was to evaluate the effect of ozonation as pretreatment on the removal of dissolved or biodegradable organic matter(DOM or BOM), the variance of DOM fractionation, and microbial regrowth by pilot-scale granular activated carbon processes in which adsorption and biodegradability was proceeding due to long time operation. Regardless of point of ozonation applied, GAC processes with ozonation(i.e., Ozonation combined with GAC Filter-adsorber; Pre O$_3$ + F/A, Ozonation combined with GAC adsorber; Post O$_3$ + GAC) compared with GAC processes without ozonation(i.e., GAC Filter-adsorber; F/A, GAC adsorber; GAC) removed approximately 10 to 20% more of DOC, hydrophilic DOM(HPI), BDOC and AOC after long period of operation that biological activity was assumed to happen. Ozonation was not found to have a significant effect on the removal of DOC, but caused the decrease of AOC by approximately 20%. It was found that the fixed bacterial biomass on GAC media did not show a significant difference between the GAC with ozonation and GAC without ozonation as pre-treatment, whereas the HPC of column effluent was more biostable at Post O$_3$ + GAC compared with F/A or GAC.

Applicability Evaluation of Two-stages and Dual Media Filtration System by the Small-scale Pilot Plant (이단이층 복합여과시스템의 소규모 파일롯 플랜트 적용성 평가)

  • Woo, Dal-Sik;Song, Si-Byum;Hwang, Byung-Gi
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.10 no.4
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    • pp.857-864
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    • 2009
  • This study aimed at developing the two stage and dual filtration system. It has a sand + activated carbon layer above the underdrain system and a sand layer above the middledrain system for pretreatment. When retrofitting an old filter bed or designing a new one, this technology can substitute the existing sand filter bed without requiring a new site. In order to extend the filtering duration, the upper layer of the filter bed consists of the rapid sand filtration with large particles which pre-treats and removes coarse particles and turbidity matters. The middle layer has biological activated carbon(BAC) and granular activated carbon(GAC) to eliminate dissolved organic matters, disinfection by-products precursors etc. The lower layer consists of the sand filtration for the post filtering mode. In this study, a pilot plant of two stage and dual filtration system was operated for 4 months in the S water treatment plant in Kyounggi-Do. The stability of turbidity was maintained below 1NTU. The TOC, THMFP and HAAFP were removed about 90% by two stage and dual filtration system, which is almost 2 times higher than S WTP. From analysis result of HPC along the depth of activated carbon + sand layer at 2nd stage, microorganism was mostly not detected, however, increment of HPC was shown as it becomes deeper. It indicates that growth of microorganism is occurred at activated carbon layer.

Analysis Method of N-Nitrosamines in Human Urine by LC-MS/MS System (LC-MS/MS 시스템을 이용한 소변 중 N-니트로사민류 분석법 확립)

  • Park, Na-Youn;Jung, Woong;Kho, Younglim
    • Journal of the Korean Chemical Society
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    • v.61 no.2
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    • pp.51-56
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    • 2017
  • N-nitrosamines are the nitroso compounds which are produced by nitrosation reactions of the secondary amine and nitrite under acidic conditions. Approximately 300 species of N-nitrosamine have been tested for carcinogenicity in laboratory experiments, with 90% of them demonstrated carcinogenic effects different animal species, including higher primates. In 1978, IARC classified NDMA and NDEA as Group 2A, and NDPA, NDBA, NPIP, NPYR and NMOR as Group 2B. In this study, we established pretreatment and analytical method for N-nitrosamines (NDMA, NDEA, NMEA, NDPA, NDBA, NPIP, NPYR and NMOR) in human urine for biological monitoring of N-nitrosamines. The analytes were extracted using solid phase extraction (SPE), then quantitative analysis was performed by LC-(APCI)-MS/MS. The accuracies of the established method were between 85.8~108.7% and precisions were lower than 20%. The limit of detection (LOD) were between 0.0002 (NDBA) and 0.0793 (NDMA) ng/ml. The linearity obtained was satisfying for the 8 N-nitrosamines, with a coefficient of determination ($r^2$) higher than 0.999. The mean concentrations of N-nitrosamines in the urine were 2.645 mg/g creatinine for NDMA, 0.067 mg/g creatinine for NDEA, 0.009 mg/g creatinine for NMEA, 0.011 mg/g creatinine for NDBA, 0.271 mg/g creatinine for NPIP and 0.413 mg/g creatinine for NPYR. NDPA and NMOR were not detected. It can be used as a instrumental methodology for evaluation and risk assessment of human exposure to N-nitrosamines for the further research.