• Title/Summary/Keyword: PRETREATMENT

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Development of Analysis Method of Caffeine and Content Survey in Commercial Foods by HPLC (HPLC를 이용한 카페인의 분석법 개발 및 시판 식품중 함유량 조사)

  • Kim, Hee-Yun;Lee, Young-Ja;Hong, Ki-Hyoung;Lee, Chul-Won;Kim, Kil-Saeng;Ha, Sang-Chul
    • Korean Journal of Food Science and Technology
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    • v.31 no.6
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    • pp.1471-1476
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    • 1999
  • A simple and practical method for determination of caffeine in foods was developed. The analysis of caffeine was performed by reverse phase high performance liquid chromatography using a ${\mu}-Bondapak\;C_{18}$ column at isocratic condition with methanol-acetic acid-water(20 : 1 : 79) on UV detector at 280 nm. The clean-up and extraction of caffeine in samples were based on a simple pretreatment using a Sep-Pak $C_{18}$ cartridge. Recovery rates obtained with this method for cider, candy, cookie, milk, ice cream and persimmon leaf tea were 99.23%, 99.50%, 99.17%, 99.37%, 98.93% and 99.10% respectively. And the detection limit of caffeine was $0.1\;{\mu}g/mL$. With this method, the range of caffeine contents extracted from coffee, green tea, black tea, Oolong tea(tea bag), soft drinks, ice cream, milk and commercial confectionery were $3.38{\sim}37.50\;mg/g,\;16.30{\sim}26.10\;mg/g,\;10.80{\sim}16.65\;mg/g,\;11.25\;mg/g,\;0.06{\sim}0.11\;mg/g,\;0.04{\sim}0.44\;mg/g,\;0.04{\sim}0.39\;mg/g\;and\;0.10{\sim}1.80\;mg/g$, respectively. But caffeine was not detected in the other tea such as Acanthopanax sessiliflorum tea, Angelica gigas tea, Angelica tea, Arrow root tea, Duchu'ng tea, Dunggulle tea, Ganoerma lucidum tea, Ginger tea powder, Persimmon leaf tea, Ssanghwa tea and Cocoa mix powder.

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Activation of Pro-Apoptotic Multidomain Bcl-2 Family Member Bak and Mitochondria-Dependent Caspase Cascade are Involved in p-Coumaric Acid-Induced Apoptosis in Human Jurkat T Cells (p-Coumaric acid에 의해 유도되는 인체 Jurkat T 세포의 에폽토시스 기전)

  • Lee, Je-Won;Kim, Young-Ho
    • Journal of Life Science
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    • v.21 no.12
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    • pp.1678-1688
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    • 2011
  • The apoptogenic effect of p-coumaric acid, a phenolic acid found in various edible plants, on human acute leukemia Jurkat T cells was investigated. Exposure of Jurkat T cells to p-coumaric acid (50-$150{\mu}M$) caused cytotoxicity and TdT-mediated dUTP nick-end labeling (TUNEL)-positive apoptotic DNA fragmentation along with Bak activation, ${\Delta}{\psi}m$ loss, activation of caspase-9, -3, -7, and -8, and PARP degradation in a dose-dependent manner. However,these apoptotic events were completely abrogated in Jurkat T cells overexpressing Bcl-2.Under these conditions, necrosis was not accompanied. Pretreatment of the cells with the pan-caspase inhibitor (z-VAD-fmk) could prevent p-coumaric acid-induced sub-$G_1$ peak representing apoptotic cells, whereas it failed to block ${\Delta}{\psi}m$ loss, indicating that the activation of caspase cascade was prerequisite for p-coumaric acid-induced apoptosis as a downstream event of ${\Delta}{\psi}m$ loss. FADD- and caspase-8-positive wild-type Jurkat T cell clone A3, FADD-deficient Jurkat T cell clone I2.1, and caspase-8-deficient Jurkat T cell clone I9.2 exhibited similar susceptibilities to the cytotoxicity of p-coumaric acid, excluding an involvement of Fas/FasL system in triggering the apoptosis. The apoptogenic activity of p-coumaric acid is more potent in malignant Jurkat T cells than in normal human peripheral T cells. Together, these results demonstrated that p-coumaric acid-induced apoptogenic activity in Jurkat T cellswas mediated by Bak activation, ${\Delta}{\psi}m$ loss, and subsequent activation of multiple caspases such as caspase-9, -3, -7, and-8, and PARP degradation, which could be regulated by anti-apoptotic protein Bcl-2.

Reactive Oxygen Species Mediates Lysophosphatidic Acid-induced Migration of SKOV-3 Ovarian Cancer Cells (SKOV-3 난소암 세포주에서 lysophosphatidic acid 유도 세포의 이동에 있어 활성산소의 역할)

  • Kim, Eun Kyoung;Lee, Hye Sun;Ha, Hong Koo;Yun, Sung Ji;Ha, Jung Min;Kim, Young Whan;Jin, In Hye;Shin, Hwa Kyoung;Bae, Sun Sik
    • Journal of Life Science
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    • v.22 no.12
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    • pp.1621-1627
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    • 2012
  • Cell motility plays an essential role in many physiological responses, such as development, immune reaction, and angiogenesis. In the present study, we showed that lysophosphatidic acid (LPA) modulates cancer cell migration by regulation of generation of reactive oxygen species (ROS). Stimulation of SKOV-3 ovarian cancer cells with LPA strongly promoted migration. but this migration was completely blocked by pharmacological inhibition of phosphatidylinositol 3-kinase (PI3K)/Akt signaling pathway. Inhibition of the ERK pathway had no effect on migration. Stimulation of SKOV-3 ovarian cancer cells with LPA significantly induced the generation of ROS in a time-dependent manner. LPA-induced generation of ROS was significantly blocked by pharmacological inhibition of PI3K or Akt, but inhibition of the ERK signaling pathway had little effect. LPA-induced generation of ROS was blocked by pretreatment of SKOV-3 ovarian cancer cells with an NADPH oxidase inhibitor, whereas inhibition of xanthine oxidase, cyclooxygenase, or mitochondrial respiratory chain complex I had no effect. Scavenging of ROS by N-acetylcysteine completely blocked LPA-induced migration of SKOV-3 ovarian cancer cells. Inhibition of NADPH oxidase blocked LPA-induced migration whereas inhibition of xanthine oxidase, cyclooxygenase, or mitochondrial respiratory chain complex I did not affect LPA-induced migration of SKOV-3 ovarian cancer cells. Given these results, we suggest that LPA induces ROS generation through the PI3K/Akt/NADPH oxidase signaling axis, thereby regulating cancer cell migration.

Histological studies on in vitro Propagation of Pulsatilla koreana Nakai (할미꽃 기내증식(器內增殖)에 관(關)한 조직학적(組織學的) 연구(硏究))

  • Lee, Man-Sang;Oh, Ki-Hong
    • Korean Journal of Medicinal Crop Science
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    • v.1 no.2
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    • pp.137-157
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    • 1993
  • This study was carried out to investigate the optimal condition for multiple propagation through leaf tissue culture and to apply anther culture techniques to Pulsatilla koreana Nakai breeding. Leaf and anther of Pulsatilla koreana Nakai were cultured on MS, MT, LS and $B_5$ media supplemented with several growth regulators and nitrogen sources under various conditions. For callus induction and differentiation from the Pulsatilla koreana leaf segments were more effective in the combination of zeatin and auxin than auxin alone. The color of the callus was green when treated with IBA alone. Shoot differentiation was more effective when treated with zeatin than auxin alone, especially the best hormoal combination for shoot differentiation was zeatin 1.0mg/l +NAA 0.1mg/l, while 2,4-D inhibited shoot differentiation. The appeared rate of S pollen was 35% in vivo, while that of S pollen by low temperature$(4^{\circ}C)$ pretreatment for 4 days was increased by 53% and the optimum culture time for callus induction from anther was uni-nucleate stage. $B_5$ basal medium supplemented with NAA 0.5mg/l and zeatin 1 mg/l was the most effective on callus formation and the best results of plant regeneration were obtained from combination of NAA 0.5mg/l and zeatin 0.5mg/l in anther culture. $NH_{4}NO_3$ as more effectives as the nitrogen source than $KNO_3$ and the combination with zeatin 2.0mg /L was the best effective. The best combination for plant regeneration in callus induced from anther was $NH_{4}NO_3$ 1650mg/l + $KNO_3$ 3800mg/l + zeatin 2.0mg/l. Ploidy level of anther-derived plants appeared 28% haploid, 47% diploid and the others were triploid, tetraploid and mixploid. In compare with E.S.T, M.D.H and P.X banding patterns were distinguished among callus, haploid and diploid plants in electrophoresis.

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Effect of Pretreatment Conditions on Effective Components of Extracts from Safflower (Carthamus tinctorius L.) Seed (전처리조건이 홍화씨 추출물의 유효성분 함량에 미치는 영향)

  • Kim, Jun-Han;Park, Jun-Hong;Kim, Jong-Kuk;Lee, Jin-Man;Moon, Kwang-Deog
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.31 no.3
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    • pp.367-372
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    • 2002
  • In order to utilize safflower seed effectively as a food material, it was processed at the conditions including roasting temperature/time of 170$\^{C}$/10 min to 210$\^{C}$/30 min, ethanol concentration of 0 to 100% (V/V) and enzyme hydrolysis with $\alpha$-amylase, $\beta$-amylase, amyloglucosidase and cellulase. Safflower seed extracts had the highest soluble solid content at the condition of 60% ethanol concentration, roasting at 190$\^{C}$ for 20 min and hydrolysis with amyloglucosidase. Total phenolic compounds increased with the ethanol concentration, showing the highest at the condition of 80% ethanol, roasting at 170$\^{C}$ for 30 min and hydrolysis with amyloglucosidase. High level total flavonoid was observed at the condition of 80% ethanol, roasting at 210$\^{C}$ for 30 min and hydrolysis with amyloglucosidase. Safflower seed had sucrose as major free sugar as well as xylose and arabinose as minor free sugars. Organic acids in safflower seed included oxalic, citric, magic and fumaric acid. Serotonin I (N-[2-(5-hydroxy-1H-indo-1-3-yl)ethyl]ftrulamide) and serotonin II (N-[2-(5-hydroxy-1H-indol-3yl)ethyl]-p-coumaramide) as antioxidant compounds increased with ethanol concentration, showing the highest revel at 60% ethanol. Acacetin content increased with temperature and roasting time, with a maximum of 69.47 mg% at 210$\^{C}$ for 30 min.

Isolation and Identification of Agarose-degrading Bacterium, Pseudoalteromonas sp. GNUM08122 (아가로오스 분해세균인 Pseudoalteromonas sp. GNUM08122 분리 및 동정)

  • Kim, Yu-Na;Jeong, Yeon-Kyu;Kim, Mu-Chan;Kim, Sung-Bae;Chang, Yong-Keun;Chi, Won-Jae;Hong, Soon-Kwang;Kim, Chang-Joon
    • Microbiology and Biotechnology Letters
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    • v.40 no.1
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    • pp.1-9
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    • 2012
  • This study's aim was to isolate microorganisms producing agarase with a high activity, with possible applications in improving the performance of the pretreatment processes for bioethanol production. Marine algaes were collected from the south coast of Korea, from which three kinds of microorganisms were isolated. After a 4-day culture of these strains at $25^{\circ}C$, crude enzymes were obtained from culture supernatant or cell-free extract by ammonium sulfate precipitation and membrane dialysis. Agarase activity was observed in these crude enzymes. Notably higher specific activity was observed in the crude enzyme obtained from the culture supernatant rather than that from the cell-free extract. This indicates that a secreted enzyme has a much greater activity than a cellular enzyme. Crude enzymes from the GNUM08122 strain were inferred to have ${\alpha}$-agarase activity because release of p-nitrophenol was observed, possibly due to the cleavage of p-nitrophenyl-${\alpha}$-D-galactopyranoside. The 16S rRNA sequence of GNUM08122 showed a close relationship to Pseudoalteromonas issachenkonii KMM 3549 (99.8%) and Pseudoalteromonas tetraodonis IMA 14160 (99.7%), which led us to assign it to the genus Pseudoalteromonas. Biochemical and physiological study revealed that this strain can grow well at $40^{\circ}C$ under a wide range of pH (pH 4~8) in high-salt conditions (10% NaCl).

Effect of xylobiose-sugar mixture on defecation frequency and symptoms in young women with constipation (자일로바이오스를 함유한 설탕이 젊은 여성의 변비 개선에 미치는 효과)

  • Lee, Jung-Sug;Kim, A-Reum;Nam, Hye-kyoung;Kyung, Myungok;Jo, Sung-Eun;Chang, Moon-Jeong
    • Journal of Nutrition and Health
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    • v.50 no.1
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    • pp.41-52
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    • 2017
  • Purpose: The objective of the study was to investigate the effects of xylobiose-sugar mixture intake on defecation frequency and constipation symptoms in 31 young women with constipation. Methods: Thirty-one subjects were assigned to two groups, and subjects in each group were administered 10 g of a 7% xylobiose-sugar mixture (Experiment 1: XBS, n = 15) or 10 g of a 7% xylobiose-sugar mixture containing coffee mix (Experiment 2: XBS coffee mix, n = 16) twice per day for 6 weeks. During the study, clinical efficacy was assessed by a daily diary record. The subjects recorded their defecation frequency and fecal characteristics. Results: During pretreatment week, mean defecation frequency of XBS subjects was 2.13 times/week, whereas that of XBS coffee mix subjects was 1.56 times/week. The mean defecation frequencies of XBS and XBS coffee mix subjects increased significantly to 3.73 times/week (p < 0.05) and 3.56 times/week by week 6 (p < 0.05), respectively. After treatment with either XBS or XBS coffee mix, patients presented significant improvements in their amounts of stool, feelings of residual stool leftness, and abdominal pain symptoms (p < 0.05). The total constipation scoring system (CSS) for diagnosing constipation symptoms significantly decreased in the XBS group (10.53 score vs 7.22 score) and in the XBS coffee mix group (10.75 score vs 6.51 score) after 6 weeks. Improvement due to intake of 7% xylobiose-containing sugar seemed to last during the experimental period. Conclusion: The addition of approximately 7% xylobiose to commercially available sweeteners has been shown to improve constipation.

Electrofusion of Yeast Cells and Their Genetic Analysis Using RAPD-PCR (효모세포의 전기융합 및 융합세포의 RAPD-PCR을 이용한 유전적 분석)

  • Kim, Seung;Kim, Jae-Sung;Sapkota, Kumar;Park, In-Sung;Cho, Moon-Gu;Park, Yeal;Chun, Hong-Sung;Choi, Bong-Suk;Park, Se-Eun;Choi, Han-Suk;Kim, Myung-Kon;Kim, Sung-Jun
    • Applied Biological Chemistry
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    • v.49 no.3
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    • pp.186-191
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    • 2006
  • In the present study, an attempt has been made to produce hybrid yeast strains of different useful and dominant characteristics. The hybrid yeast strains were produced by electrofusion and their genetic analysis were performed by RAPD-PCR (random amplified polymorphic DNA-polymerase chain reaction). The protoplast of Saccharomyces cerevisiae KCTC 7904 and Zygosaccharomyces rouxii KCTC 7966 were obtained above 92% when treated with lyticase at $30^{\circ}C$ for $60{\sim}90$ min after the pretreatment of $1{\sim}2%$ 2-mercaptoethanol at $30^{\circ}C$ for $15{\sim}20$ min. The fusant was produced from paired protoplast stage under the electric pulse at high frequency conditions (1.5 MHz/50 pV, 615 $V/256\;{\mu}sec$) within glass-platinum made electrofusion chamber. Changes in RAPD patterns in mother cells and hybrid cells proved that the fusant contains two types of yeast gene originated from its parent. Furthermore, fermentation characters exhibits by the fusant cell confirmed its genetic changes. These results suggest that genetically stable hybrid yeast strains of economic importance can be produced by electrofusion technique and these electrofused yeast cells have an enormous impact in biotechnology and biomedicine.

Preliminary Results of Phase I/II Study of Simultaneous Modulated Accelerated (SMART) for Nasopharyngeal Carcinoma (비인강암의 치료에서 동시차등조사가속치료를 적용한 제1상/2상 연구의 예비적 결과)

  • Park Jin-Hong;Back Geum-Mun;Yi Byong-Yong;Choi Eun-Kyung;Ahn Seung-Do;Shin Seong-Soo;Kim Jung-Hun;Kim Sang-Yoon;Lee Bong-Jae;Nam Soon-Yuhl;Choi Seung-Ho;Kim Seung-Bae;Lee Sang-Wook
    • Radiation Oncology Journal
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    • v.24 no.1
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    • pp.1-10
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    • 2006
  • Puroose: To present preliminary results of intensity-modulated radiotherapy (IMRT) using the simultaneous modulated accelerated radiation therapy (SMART) boost technique in patients with nasopharyngeal carcinoma (NPC). Materials and Methods: Twenty patients who underwent IMRT for non-metastatic NPC at the Asan Medical Center between September 2001 and December 2003 were prospectively evaluated. IMRT was delivered using the 'step and shoot' SMART technique at prescribed doses of 72 Gy (2.4 Gy/day) to the gross tumor volume (GTV), 60 Gy (2 Gy/day) to the clinical target volume (CTV) and metastatic nodal station, and 46 Gy (2 Gy/day) to the clinically negative neck region. Eighteen patients also received concurrent chemotherapy using cisplatin once per week. Results: The median follow-up period was 27 months. Nineteen patients completed the treatment without interruption; the remaining patient interrupted treatment for 2 weeks owing to severe pharyngitis and malnutrition. Five patients (25%) had RTOG grade 3 mucositis, whereas nine (45%) had grade 3 pharyngitis. Seven patients (35%) lost more than 10% of their pretreatment weight, whereas 11 (55%) required intravenous fluids and/or tube feeding. There was no grade 3 or 4 chronic xerostomia. All patients showed complete response. Two patients had distant metastases and loco-regional recurrence, respectively. Conclusion: IMRT using the SMART boost technique allows parotid sparing, as shown clinically and by dosimetry, and may also be more effective biologically. A larger population of patients and a longer follow-up period are needed to evaluate ultimate tumor control and late toxicity.

7-Ketocholesterol Induces Vascular Smooth Muscle Cell Apoptosis via Akt Degradation (7-Ketocholesterol에 의한 Akt 감소와 혈관평활근세포의 세포자멸사)

  • Seo, Kyo Won;Kim, Chi Dae;Lee, Won Suk
    • Journal of Life Science
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    • v.26 no.2
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    • pp.226-233
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    • 2016
  • Vascular smooth muscle cell (VSMC) apoptosis has been identified in various vascular diseases, including atherosclerosis and restenosis after angioplasty, and has been known to precipitate atherosclerotic plaque instability and rupture. Oxysterols are known as inducers of apoptosis in VSMC, and 7-ketocholesterol (7KC) is the major nonenzymically formed oxysterol in atherosclerotic lesions. The precise mechanism underlying VSMC apoptosis is still poorly understood. In this study, we investigated whether 7KC causes apoptosis, and characterized its apoptotic mechanisms in primary cultured rat aortic VSMC. Cell viability was assessed by MTT assay and trypan blue assay. Apoptosis was assessed by flow cytometry, immunofluorescence, immunoprecipitation, and Western blot analyses. 7KC markedly decreased the VSMC viability in a time- and concentration-dependent manner, and increased the production of 4-hydroxynonenal (HNE), a major end-product of lipid peroxidation, which also decreased the VSMC viability. Pretreatment with 2,4-dinitrophenylhydrazine, a well-known reagent of lipid peroxidation-derived aldehydes, significantly restored the 7KC-decreased viability of VSMC. Furthermore, HNE, as well as 7KC, reduced the level of total Akt, a major mediator of cell survival. The 7KC-decreased level of total Akt was significantly restored by pretreatments with 2,4-dinitrophenylhydrazine and N-acetylcysteine. Lactacystin, a proteasome inhibitor, protected VSMC against apoptosis and Akt degradation, but did not inhibit HNE production. In the immunoprecipitation assay, 7KC increased HNE-modified Akt. From the results, it seems that, in atherosclerotic lesions, 7KC induces HNE production in VSMC, and this HNE binds to Akt, proceeding to proteasomal degradation of Akt, through which mechanism the atherosclerotic plaque instability may be facilitated.