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Determination of plasma C16-C24 globotriaosylceramide (Gb3) isoforms by tandem mass spectrometry for diagnosis of Fabry disease (패브리병(Fabry) 진단을 위한 혈장 중 Globotriaosylceramide (Gb3)의 탠덤매스 분석법 개발과 임상 응용)

  • Yoon, Hye-Ran;Cho, Kyung-Hee;Yoo, Han-Wook;Choi, Jin-Ho;Lee, Dong-Hwan;Zhang, Kate;Keutzer, Joan
    • Journal of Genetic Medicine
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    • v.4 no.1
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    • pp.45-52
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    • 2007
  • Purpose : A simple, rapid, and highly sensitive analytical method for Gb3 in plasma was developed without labor-ex tensive pre-treatment by electrospray ionization MS/ MS (ESI-MS/MS). Measurement of globotriaosy lceramide (Gb3, ceramide trihex oside) in plasma has clinical importance for monitoring after enzyme replacement therapy in Fabry disease patients. The disease is an X-linked lipid storage disorder that results from a deficiency of the enzyme ${\alpha}$-galactosidase A (${\alpha}$-Gal A). The lack of ${\alpha}$-Gal A causes an intracellular accumulation of glycosphingolipids, mainly Gb3. Methods : Only simple 50-fold dilution of plasma is necessary for the extraction and isolation of Gb3 in plasma. Gb3 in diluted plasma was dissolved in dioxane containing C17:0 Gb3 as an internal standard. After centrifugation it was directly injected and analyzed through guard column by in combination with multiple reaction monitoring mode of ESI-MS/MS. Results : Eight isoforms of Gb3 were completely resolved from plasma matrix. C16:0 Gb3 occupied 50% of total Gb3 as a major component in plasma. Linear relationship for Gb3 isoforms w as found in the range of 0.001-1.0 ${\mu}g$/mL. The limit of detection (S/N=3) was 0.001 ${\mu}g$/mL and limit of quantification was 0.01 ${\mu}g$/mL for C16:0 Gb3 with acceptable precision and accuracy. Correlation coefficient of calibration curves for 8 Gb3 isoforms ranged from 0.9678 to 0.9982. Conclusion : This quantitative method developed could be useful for rapid and sensitive 1st line Fabry disease screening, monitoring and/or diagnostic tool for Fabry disease.

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Nutritional Composition and Cytoprotective Effect of Moringa oleifera Lam. (Drumstick-tree (Moringa oleifera Lam.)의 주요 영양성분 및 추출물의 신경세포 보호 효과)

  • Jin, Su Il;Kim, Hyeon Ju;Jeong, Ji Hee;Jin, Dong Eun;Choi, Sung-Gil;Heo, Ho Jin
    • Korean Journal of Food Science and Technology
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    • v.46 no.5
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    • pp.609-615
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    • 2014
  • The cytoprotective effect of Moringa oleifera Lam. (drumstick tree) on neuronal cells was investigated to confirm the physiological benefits associated with this natural food resource. First, the drumstick tree extract was chemically analyzed to determine inherent nutritional constituents. Calcium and potassium were identified as the major mineral constituents, and palmitic acid (C16:0, 16.33%) and gadoleic acid (C20:01, 66.34%) were detected as the major fatty acids. Moreover, drumstick tree extract contained 94.78 mg/100 g vitamin E and 112.61 mg/100 g niacin. PC12 cells were used to study the cytoprotective effects of drumstick tree extract. Intracellular accumulation of reactive oxygen species was significantly reduced when $H_2O_2$ treated-neuronal cells were cultured in a medium containing the methanolic extract of drumstick tree, compared to cells treated with only $H_2O_2$. Cell viability assay using MTT showed that the extract protected cells against $H_2O_2$-induced neurotoxicity and inhibited LDH leakage from the cell membrane. Caspase assay showed that the extract exerted cytoprotective effect against apoptosis. Consequently, these data suggest that drumstick tree is a useful natural resource with positive effects on human health.

Protective Effect of Radiation-induced New Blackberry Mutant γ-B201 on H2O2-induced Oxidative Damage in HepG2 Cells (H2O2 에 의해 유도된 HepG2 세포의 산화적 스트레스에 대한 신품종 방사선 돌연변이 블랙베리 γ-B201의 세포 보호 효과)

  • Cho, Byoung Ok;Lee, Chang-Wook;So, Yangkang;Jin, Chang-Hyun;Yook, Hong-Sun;Byun, Myung-Woo;Jeong, Yong-Wook;Park, Jong Chun;Jeong, Il-Yun
    • Korean Journal of Food Science and Technology
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    • v.46 no.3
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    • pp.384-389
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    • 2014
  • The objective of the present study was to investigate the chemical composition of anthocyanin-enriched extract of radiation-induced blackberry (Rubus fruticosus L.) mutant (${\gamma}$-B201) as well as the protective effect of ${\gamma}$-B201 against oxidative stress in vitro. The cytotoxicity, reactive oxygen species (ROS) scavenging capacity, and DNA damage were assessed by WST-1 assay, flow cytometry, and comet assay, respectively. Lactate dehydrogenase, superoxide dismutase, and catalase activities were determined by using a commercial kit. The in vitro results showed that ${\gamma}$-B201 increased the cell viability, reduction of lactate dehydrogenase release, and intracellular ROS scavenging capacity in hydrogen peroxide ($H_2O_2$)-treated HepG2 cells. Furthermore, treatment with ${\gamma}$-B201 attenuated DNA damage in $H_2O_2$-treated HepG2 cells and treatment with ${\gamma}$-B201 restored the activity of superoxide dismutase and catalase in $H_2O_2$-treated HepG2 cells. In conclusion, the present study suggests that ${\gamma}$-B201 blackberry extract can exert a significant cytoprotective effect against $H_2O_2$-induced cell damage.

Effect of gomchwi (Ligularia fischeri) extract against high glucose- and H2O2-induced oxidative stress in PC12 cells (PC12 신경세포에서 고당 및 과산화수소로 유도된 산화적 스트레스에 대한 곰취 추출물의 효과)

  • Park, Sang Hyun;Park, Seon Kyeong;Ha, Jeong Su;Lee, Du Sang;Kang, Jin Yong;Kim, Jong Min;Lee, Uk;Heo, Ho Jin
    • Korean Journal of Food Science and Technology
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    • v.48 no.5
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    • pp.508-514
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    • 2016
  • Effects of the ethyl acetate fraction from gomchwi (Ligularia fischeri) extract against high $glucose/H_2O_2-induced$ oxidative stress and in vitro neurodegeneration were investigated to confirm the physiological property of the extract. The ethyl acetate fraction of gomchwi extract showed the highest total phenolic contents than the other solvent fractions. An anti-hyperglycemic activity of the ethyl acetate fraction was evaluated using the ${\alpha}-glucosidase$ inhibitory assay, and the half maximal inhibitory concentration ($IC_{50}$) value for ${\alpha}-glucosidase$ was found to be $727.64{\mu}g/mL$. In addition, the ethyl acetate fraction showed excellent 2,2'-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) diammonium salt radical scavenging activity, and inhibition of malondialdehyde production. The ethyl acetate fraction also decreased intracellular reactive oxygen species, whereas neuronal cell viability against high glucose/$H_2O_2$-induced cytotoxicity was found to be increased. Finally, 3,5-dicaffeoylquinic acid as a main phenolic compound in the ethyl acetate fraction was analyzed by high-performance liquid chromatography. These results suggest that gomchwi might be a good natural source of functional materials to prevent diabetic neurodegeneration.

Comparison of Anti-oxidative Activity in a Single Serving Size of the Commercial Coffees and Teas

  • Kim, Tae-Hun;Lee, Seulgi;Seo, Jin Woo;Bing, Sun Hye;Kim, Jong Im;Kwon, Eui-Ra;Jo, Gune-Hee;Lee, Jae-Myean;Choi, Joon Sig
    • Journal of Food Hygiene and Safety
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    • v.32 no.6
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    • pp.460-469
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    • 2017
  • The aim of this work was to study the comparison of anti-oxidative activity in a single serving size of commercial coffees and teas. Commercial regular coffees and teas, including, brand regular coffees ($BC_A$, $BC_B$, $BC_C$, $BC_D$, and $BC_E$), green tea ($GT_A$, $GT_B$, $GT_C$, and $GT_D$), black tea ($BT_A$, $BT_B$, and $BT_C$), pu-erh tea ($PT_A$, $PT_B$, and $PT_C$), chamomile tea ($CT_A$, $CT_B$, and $CT_C$), peppermint tea ($P_A$, $P_B$, and $P_C$), polygonatum odoratum tea ($POT_A$, $POT_B$, and $POT_C$), and jujube tea ($JT_A$, $JT_B$, and $JT_C$) were assayed for the levels of ascorbic acid, caffeine, total content of polyphenols and flavonoids, and ability to scavenge free radicals, using two in vitro antioxidant assays. The scavenging abilities of $BC_A$ and $BC_C$ were $664.91{\pm}48.87mg$ ascorbic acid equivalent/serving size and $624.36{\pm}16.18mg$ ascorbic acid equivalent/serving size, respectively. The four beverage samples ($BC_A$, $BC_C$, $GT_D$, and $BT_A$) significantly reduced the production of reactive oxygen species (ROS) and intracellular oxidative stress induced by $H_2O_2$. These results suggest that the beverages possess significant radical scavenging ability, which may be due to the presence of antioxidants. Furthermore, the significant reducing level of ROS evidences the potential antioxidant effects of these beverages in human cells.

Production of Fungal Lipids - I. On Intracellular Fungal Lipids - (곰팡이 유지 생산에 관한 연구 - 제 1 보 : 곰팡이 세포내 유지에 대하여 -)

  • Yoo, Jin-Young;Shin, Dong-Hwa;Yim, Ho;Min, Byong-Yong;Suh, Kee-Bong
    • Korean Journal of Food Science and Technology
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    • v.12 no.2
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    • pp.97-102
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    • 1980
  • Cladosporium fulvum, Aspergillus ochraceus, Aspergillus terreus and N-1 (unidentified species) were cultured on the artificial media containing sucrose as a carbon source at 20, 25 and $30^{\circ}C$ for 10 to 12 days. The lipids in the felts were extracted with chloroform-methanol mixture and the class composition and fatty acids of the lipid were determined. The summarized results are as follows 1. The average felts produced by each species per 100 ml of media were $3.82{\pm}0.30g$ for Cl. fulvum, $2.62{\pm}0.23g$ for Asp. ochraceus, $4.24{\pm}0.25g$ for Asp. terreus and $4.62{\pm}0.10g$ for N-1. Their crude fat contents $27.5{\pm}1.61%,\;50.47{\pm}1.00%,\;46.6{\pm}1.59%$ and 33.78 % and the fat coefficient 6.92, 8.88, 13.01 and 10.28, respectively. 2. The lipids produced by these species were mainly composed of triglyceride and the next free fatty acid in Cl. fulvum and N-1 and phospholipid Asp. ochraceus and Asp. terreus. 3. The major fatty acids of the lipids were in order of oleic, palmitic, linoleic and stearic acids in Asp. ochraceus, Asp. terreus and Cl. fulvum and linoleic, palmitic, oleic and stearic acid in N-1. The total percentage contents of these major fatty acids were over 98 % the former and over 95 % the latter. 4. The constituent fatty acids of the lipid were changed depending on the incubation temperature but hardly found a certain tendency except linoleic acid which was higher at lower temperature. 5. The total percentages of unsaturated fatty acids in the lipids were $50{\sim}60%$ and comparatively higher at lower incubation temperature.

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Protective Effects of Sea Buckthorn (Hippophae rhamnoides L.) Leaves Fermented with Hericium erinaceum Mycelium against Oxidative Modification of Biological Macromolecules and Cell Death (노루궁뎅이 버섯균사체를 이용한 비타민나무 발효물이 생체고분자의 산화적 변형과 세포사멸에 미치는 보호 영향)

  • Kim, Seung-Sub;Kyeong, Inn-Goo;Lee, Mi-La;Kim, Dong-Goo;Shin, Ji-Young;Yang, Jin-Yi;Lee, Gwang-Ho;Eum, Won-Sik;Kang, Jung-Hoon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.1
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    • pp.35-43
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    • 2015
  • In this study, hot water extract from sea buckthorn (Hippophae rhamnoides L.) leaves fermented with Hericium erinaceum mycelium (SBT-HE) was assessed for protection against oxidative modification of biological macromolecules and cell death. Antioxidant activity of SBT-HE was evaluated based on 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical, and peroxyl radical scavenging assays. SBT-HE showed 65.06% DPPH radical scavenging activity at $500{\mu}g/mL$, 98.83% ABTS radical scavenging activity at $50{\mu}g/mL$, and 44.03% peroxyl radical scavenging activity at $100{\mu}g/mL$. SBT-HE significantly inhibited DNA strand breakage induced by peroxyl radical. SBT-HE also prevented peroxyl radical-mediated human serum albumin modification. SBT-HE effectively inhibited $H_2O_2$-induced cell death and significantly increased cell survival by 21.59% at $100{\mu}g/mL$. SBT-HE also reduced intracellular reactive oxygen species levels in $H_2O_2$-treated cells. The results suggest that SBT-HE can contribute to antioxidant activity and protect cells from oxidative stress-induced cell injury.

Antibacterial and Intracellular Clearance Effect of Saururus chinensis Baill Water Extract against Salmonella typhimurium (삼백초 수용성 추출물의 Salmonella typhimurium 균에 대한 항균 및 숙주세포 내 사멸 효과 규명)

  • Kim, Dong-Hyeok;Lee, Jin-Ju;Lim, Jeong-Ju;Kim, Dae-Geun;Kim, Gon-sub;Lee, Hu-Jang;Min, Won-gi;Rhee, Man-Hee;Chang, Hong-Hee;Kim, Suk
    • Journal of agriculture & life science
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    • v.44 no.6
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    • pp.111-119
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    • 2010
  • Salmonella spp. are one of major pathogens for zoonosis in worldwide, and can replicate within host cells and generally cause enterocolitis and foodborne poisoning which represents a considerable public health burden. The present study was designated to investigate the safty for host cells, antibacterial effects of Saururus chinensis Baill water extract (SCWE) on pure culture and infection with Salmonella enterica serovar Typhimurium (S. typhimurium) in murine derived macrophage RAW 264.7 cells. The different treatment of SCWE concentration (1, 10 or $100{\mu}g/ml$) did not show any cytotoxic effect to RAW 264.7 cells for 24 h incubation. In determination of antibacterial activity of SCWE against S. typhimurium, bacterial viability was markedly decreased compared to SCWE-untreated control. In RAW 264.7 cells, SCWE significantly induced morphological change (p<0.05). In infection assay of S. typhimurium in RAW 264.7 cells pretreated with $100{\mu}g/ml$ of SCWE, which are non-cytotoxic concentration, bacterial uptake ability of macrophage was increased corresponding with morphological change, whereas bacterial survival rates within macrophage was markedly reduced comparing to that of SCWE-untreated control. Furthermore, nitric oxide (NO) production in SCWE-treated cells was slightly decreased until 24 h post infection. Taken together, these findings demonstrated that SCWE have the antibacterial activity for S. typhimurium and the protective effects against S. typhimurium infection through activating murine macrophage independent on NO, suggesting that SCWE were beneficial on the disease caused by intracellularly replicating pathogens as a safe alternatives of conventional chemotherapies.

Anti-inflammation effect of blueberry (Vaccinium ashei) leaf extract on RAW 264.7 macrophages stimulated by lipopolysaccharide (Lipopolysaccharide에 의해 활성화된 RAW 264.7대식세포에서 블루베리 잎(Vaccinium ashei) 추출물의 항염증 효과)

  • Kim, Dong In;Kim, Hyun Jung;Yun, Jong Moon;Lee, Ji Hye;Han, So Jung;Kim, Ha Eun;Jang, Min Jung;An, Bong Jeun
    • Food Science and Preservation
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    • v.25 no.1
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    • pp.107-116
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    • 2018
  • The aim of this study is to investigate the antioxidant and intracellular anti-inflammatory efficacy of blueberry leaf extracted with hot water (BLW), 70% ethanol (BLE), and 70% acetone (BLA) in RAW 264.7 macrophages. In order to evaluate the anti-inflammatory effect of blueberry leaf extracts, RAW 264.7 macrophages were stimulated with lipopolysaccharide (LPS) to induce the production of inflammation-related factors, which were measure by Western blotting and real-time PCR methods. i-NOS, COX-2 protein, and mRNA expression showed concentration-dependent decrease. The decreases in the mRNA expression levels of interleukin-$1{\beta}$ (IL-$1{\beta}$), interleukin-6 (IL-6), tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), and prostaglandin $E_2$ ($PGE_2$) were concentration-dependent. Further, the antioxidant effects of blueberry leaf on total polyphenol contents, electron donating ability and $ABTS^+$ radical scavenging activity were evaluated. The total polyphenol contents of BLW, BLE, and BLA were $217.04{\pm}2.98$, $156.72{\pm}3.90$, and $182.88{\pm}3.02mg\;TAE/g$, respectively, while the electron donating abilities at $1,000{\mu}g/mL$ of BLW, BLE, and BLA were 81.7, 79.6, and 79.3%, respectively. The $ABTS^+$ radical scavenging activity was fond to be concentration dependent. The nitric oxide (NO) production inhibition activities at $50{\mu}g/mL$ of BLW, BLE, and BLA were 35.1, 42.4 and 42.7%, respectively. In conclusion, the antioxidant and anti-inflammatory test results indicate that blueberry leaf extracts (BLW, BLE, and BLA) can be used as potential anti-inflammatory agents.

Protective effect of Codium fragile extract on fine dust (PM2.5)-induced toxicity in nasal cavity, lung, and brain cells (미세먼지(PM2.5)로 유도된 세포(비강, 폐, 뇌)독성에 대한 청각(Codium fragile)의 보호효과)

  • Kim, Gil Han;Park, Seon Kyeong;Kang, Jin Yong;Kim, Jong Min;Shin, Eun Jin;Moon, Jong Hyeon;Kim, Min Ji;Lee, Hyo Lim;Jeong, Hye Rin;Heo, Ho Jin
    • Korean Journal of Food Science and Technology
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    • v.53 no.2
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    • pp.223-229
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    • 2021
  • To evaluate the protective effect of Codium fragile on fine dust (PM2.5)-induced cytotoxicity, we investigated its antioxidant activity and cell protective effect on PM2.5-exposed cells. The 40% ethanolic extract of C. fragile showed the highest total phenolic content, whereas the water extract of C. fragile showed the highest total polysaccharide content. The protective effect of the extracts on PM2.5-induced oxidative damage in nasal cavity (RPMI2650), lung (A549), brain (MC-IXC), hippocampus (HT-22), and microglia (BV-2) cells was evaluated by measuring the intracellular reactive oxygen species (ROS) content and cell viability. The results showed that the 40% ethanolic extract more efficiently inhibited ROS production than the water extract. In contrast, PM2.5-exposed cells treated with the water extract showed higher viability than those treated with the 40% ethanolic extract.