• Title/Summary/Keyword: Escherichia coli

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Detection of Pathogenic Yersinia Enterocolitica in Drinking Water and Vegetables by Mutiplex-PCR (Multiplex-PCR에 의한 먹는샘물 및 야채류로부터의 병원성 Yersinia enterocolitica의 신속검출)

  • 이택수;박부길;오덕환
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.32 no.1
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    • pp.35-41
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    • 2003
  • The study was conducted to develope a rapid method for the detection of Yersinia enterocolitica in spring water and vegetables via multiplex polymerase chain reaction (PCR) technique using ail, yst, uirF and subgenus-specific Y16S primers. Specificity and sensitivity of multiplex PCR and application of best primers for the detection of Y. enterocolitica from spring water and vegetables were investigeted. Y. enterocolitica ATCC 27729 strains gave 356 bP and 200 bp (Y16S) and 134 bp (yst) bands. but Y. enterocolitica ATCC 9610 and ATCC 23715 strains gave 200 bp and 134 bp bands.In the meanwhile, non-pathogenic Yersinia species, such as Y. frederikseni, Y. inter-media, Y. kristenseni and Y. pseudotuberculosis gave only single 200 bp band, and other bacteria including Escherichia coli O157:H7 ATCC 25392, Shigella dysenteri. Staphylococcu aureus ATCC 25923 and Listeria mo-nocytogenes ATCC 19111 did not show any bands. Among primers, yst and Y16S primer showed the best sensitivity. Seven CFU/mL Y. enterocolitica cells could be detected with yst and Y16S primers and the sensitivity was significantly improved by the further 2nd PCR after 38 cycles of first PCR amplication. Spring water, cabbage and mushroom were inoculated with Y. enterocolitica to determine the sensitivity of multiplex-PCR for the rapid detection of Y. enterocolitica. Multiplex-PCR assay could detect 7 or 70 cells in spring water and vegetables using whole cell lysate with repeating PCR amplication.

Processed Panax ginseng, sun ginseng, inhibits the differentiation and proliferation of 3T3-L1 preadipocytes and fat accumulation in Caenorhabditis elegans

  • Lee, Hyejin;Kim, Jinhee;Park, Jun Yeon;Kang, Ki Sung;Park, Joeng Hill;Hwang, Gwi Seo
    • Journal of Ginseng Research
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    • v.41 no.3
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    • pp.257-267
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    • 2017
  • Background: Heat-processed ginseng, sun ginseng (SG), has been reported to have improved therapeutic properties compared with raw forms, such as increased antidiabetic, anti-inflammatory, and antihyperglycemic effects. The aim of this study was to investigate the antiobesity effects of SG through the suppression of cell differentiation and proliferation of mouse 3T3-L1 preadipocyte cells and the lipid accumulation in Caenorhabditis elegans. Methods: To investigate the effect of SG on adipocyte differentiation, levels of stained intracellular lipid droplets were quantified by measuring the oil red O signal in the lipid extracts of cells on differentiation Day 7. To study the effect of SG on fat accumulation in C. elegans, L4 stage worms were cultured on an Escherichia coli OP50 diet supplemented with $10{\mu}g/mL$ of SG, followed by Nile red staining. To determine the effect of SG on gene expression of lipid and glucose metabolism-regulation molecules, messenger RNA (mRNA) levels of genes were analyzed by real-time reverse transcription-polymerase chain reaction analysis. In addition, the phosphorylation of Akt was examined by Western blotting. Results: SG suppressed the differentiation of 3T3-L1 cells stimulated by a mixture of 3-isobutyl-1-methylxanthine, dexamethasone, and insulin (MDI), and inhibited the proliferation of adipocytes during differentiation. Treatment of C. elegans with SG showed reductions in lipid accumulation by Nile red staining, thus directly demonstrating an antiobesity effect for SG. Furthermore, SG treatment down-regulated mRNA and protein expression levels of peroxisome proliferator-activated receptor subtype ${\gamma}$ ($PPAR{\gamma}$) and CCAAT/enhancer-binding protein-alpha ($C/EBP{\alpha}$) and decreased the mRNA level of sterol regulatory element-binding protein 1c in MDI-treated adipocytes in a dose-dependent manner. In differentiated 3T3-L1 cells, mRNA expression levels of lipid metabolism-regulating factors, such as amplifying mouse fatty acid-binding protein 2, leptin, lipoprotein lipase, fatty acid transporter protein 1, fatty acid synthase, and 3-hydroxy-3-methylglutaryl coenzyme A reductase, were increased, whereas that of the lipolytic enzyme carnitine palmitoyltransferase-1 was decreased. Our data demonstrate that SG inversely regulated the expression of these genes in differentiated adipocytes. SG induced increases in the mRNA expression of glycolytic enzymes such as glucokinase and pyruvate kinase, and a decrease in the mRNA level of the glycogenic enzyme phosphoenol pyruvate carboxylase. In addition, mRNA levels of the glucose transporters GLUT1, GLUT4, and insulin receptor substrate-1 were elevated by MDI stimulation, whereas SG dose-dependently inhibited the expression of these genes in differentiated adipocytes. SG also inhibited the phosphorylation of Akt (Ser473) at an early phase of MDI stimulation. Intracellular nitric oxide (NO) production and endothelial nitric oxide synthase mRNA levels were markedly decreased by MDI stimulation and recovered by SG treatment of adipocytes. Conclusion: Our results suggest that SG effectively inhibits adipocyte proliferation and differentiation through the downregulation of $PPAR{\gamma}$ and $C/EBP{\alpha}$, by suppressing Akt (Ser473) phosphorylation and enhancing NO production. These results provide strong evidence to support the development of SG for antiobesity treatment.

Antibacterial Activity of Hippophae rhamnoides Leaf Extract and the Stability of a Cream with the Extract (비타민나무 잎 추출물의 항균 활성 및 크림의 안정성)

  • Chae, Kyo-Young;Kim, Jung-Eun;Park, Soo-Nam
    • Microbiology and Biotechnology Letters
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    • v.40 no.1
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    • pp.43-49
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    • 2012
  • In this study, we investigated the antibacterial activity and stability of a cream containing Hippophae rhamnoides leaf extract. The MIC values of ethyl acetate fraction from an H. rhamnoides leaf on Escherichia coli, Pityrosporum ovale, Propionibacterium acnes and Staphylococcus aureus were 0.5%, 0.25%, 0.25% and 0.06%, respectively. Stability evaluations, pH, viscosity and absorbance of the cream containing 0.25% ethyl acetate fraction of H. rhamnoides, were performed. The cream was measured under 4 different temperature conditions under sunlight at 2-week intervals for 12 weeks. The viscosity and pH were measured by a comparison of the experimental cream with a similar control cream. The H. rhamnoides extract was found to have contributed to the stability of the emulsion product via a protective effect in maintaining the viscosity of the cream against sunlight. The absorbance variations of the experimental cream at 270 nm were, under sunlight; $45^{\circ}C$, $37^{\circ}C$, $25^{\circ}C$, and $4^{\circ}C$. In addition, any change in color or smell was not observed through the 12 weeks of the experimental period. These results indicated that the cream containing 0.25% ethyl acetate fraction of H. rhamnoides leaf extract was stable. Accordingly, this suggests that further study is needed to provide additional information for manufacturers, who are seeking the application of the extract to improve anti-oxidant and antibacterial activities and the stability of cosmetic products.

Effects of γ-Irradiation from Cobalt-60 on Immunogenicity of Eimeria tenella (Cobalt-60 γ-선 조사가 Eimeria tenella 의 면역원성에 미치는 영향)

  • Youn, Hee-jeong;Kang, Yung-bai;Jang, Du-hwan
    • Korean Journal of Veterinary Research
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    • v.33 no.4
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    • pp.657-664
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    • 1993
  • To reveal the immunogenicity of ${\gamma}-irradiated$ E tenella and its progeny, a series of experiments on the effects of Cobalt-to ${\gamma}-irradiation$ was performed. The SPF chickens inoculated with diffenrt doses of inoculum were challenged with $1{\times}10^5$ oocysts of virulent E tenella. The levels of 100 Gy ${\gamma}-irradiation$ from $^{60}Co$ and of inoculum with $1{\times}10^4$ oocysts were recognized as proper as immunogen by comparison of survival rates, body weight gains, blood in feces and lesion scores in the chickens. In these trials of challenge with virulent E tenella after inoculation with $1{\times}10^4$ oocysts of the ${\gamma}-irradiated$ E tenella and its progeny, the survival rates of the chickens challenged with the virulent E tenella after immunization with the 1st and the 3rd progeny groups of ${\gamma}-irradiated$ E tenella oocysts were higher(l00%) than that(87.0%) of the challenged control group. The signs of blood in feces and the lesion scores were seen markedly lower with the ourput of the smaller number of oocysts, i.e. OPG 103,900 and 25,800 in the groups of the 1st and the 3rd progeny, respectively, than those(OPG 1,658,900) of the challenged control group. The body weight gains of the 1st and the 3rd progeny groups, the 1st week and the 2nd week after challenge, were higher (2.6g and 155.4g, 11.6g and 168.9g respectively) than those(-85.8g and 63.6g, respectively) of the challenged control group, and the feed conversion ratios(FCR 3.28 and 2.96) of the 1st and the 3rd progeny groups were lower than that(FCR 5.60) of the groups challenged control group. The anticoccidial indices(70.5 and 93.9) of the groups challenged with the virulent oocysts of E tenella after immunization with the 1st and the 3rd progeny of the ${\gamma}-irradiated$ E tenella were significantly higher than that (ACI -81.9) of the challenged control group. It was thought that the immunogenicity of ${\gamma}-irradiated$ E tenella would be increase according to increase the number of generation passaged in chicken. That might be because of increasing the pathogenicity of ${\gamma}-irradiated$ E tenella according to increase the number of generation passaged in chicken.

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Components and Antimicrobial Activity of Veiled Lady Mushroom, Dictyophora echinovolvata (흰돌기망태버섯(가칭; Dictyophora echinovolvata)의 일반성분 및 항미생물활성)

  • Cheong, Jong-Chun;Cho, Soo-Muk;Jeong, Joon-Ho;Park, Jeong-Sik;Chung, Bong-Koo;Lee, Dong-Chul
    • The Korean Journal of Mycology
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    • v.29 no.2
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    • pp.79-85
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    • 2001
  • A strain of Dictyophora echinovolvata ASI 32002 showing good fruiting body formation was selected. Analyses of chemical and nutritional components as well as antimicrobial activity of different parts of the mushroom such as mycelium, egg, and fruiting body were carried out. There were differences in the chemical compositions and the quantities depending on developmental stages of veiled lady mushroom, D. echinovolvata ASI 32002. Nitrogen, phosphate, magnesium, and calcium in inorganic chemicals were abundant in mycelium, and potassium and mineral elements were abundant in the egg and fruiting body. Mannitol and trehalose were abundant in free sugar contents. Glutamic acid and arginine in mycelium and aspartic acid and glutamic acid in egg and fruiting body were abundant in free amino acid contents. Linoleic acid, an polyunsaturated fatty acid, was abundant in all parts of the Dictyophora species, but compositions and quantities of other fatty acids varied depending on the different parts of the mushroom. It was detected that malic acid, lactic acid and acetic acid in mycelium, formic acid, acetic acid and fumaric acid in egg, and malic acid, citric acid, lactic acid, fumaric acid in fruiting body were abundant. The methanol extracts of D. echinovolvata ASI 32002 mycelium showed antifungal activity with minimal inhibition concentration (MIC) of $62{\sim}125\;{\mu}g/ml$ that was similar levels of cyclohexamide against Aspergillus awamori, Hypocrea nigricance and Trichoderma virens. The MIC of extracts from mycelium and fruiting body against Candida albicans was $250\;{\mu}g/ml$, similar to that of tetracycline. In addition to the above results, further as food additives and ingredient of cosmetics.

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Expression and Purification of Three Lipases (LipAD1, LipAD2, and LipAD3) and a Lipase Chaperone (LipBD) from Acinetobacter schindleri DYL129 (Acinetobacter schindleri DYL129 유래의 3개 lipases와 chaperone의 발현과 정제)

  • Kim, Sun-Hee;Lee, Yong-Suk;Jeong, Hae-Rin;Pyeon, Hyo-Min;You, Ju-Soon;Choi, Yong-Lark
    • Journal of Life Science
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    • v.29 no.4
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    • pp.492-498
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    • 2019
  • Previously, three kinds of lipases, lipAD1, lipAD2, and lipAD3, and lipase chaperone, lipBD, of Acinetobacter schindleri DYL129 isolated from soil sample were reported. In this report, three lipase and lipase chaperone were cloned into the pET32a(+) or pGEX-6P-1 vectors for protein expression in Escherichia coli, and named as pETLAD1, pETLAD2, pETLAD3 and pETLBD or pGEXLAD1, pGEXLAD 2, pGEXLAD3 and pGEXLBD, respectively. Protein expression rate was higher in pET system than in pGEX system. Although LipAD1 and LipAD2 were produced as inclusion bodies, their expression levels were high. So LipAD1 and LipAD2 could be solubilized in 8 M urea buffer and purified. LipAD3 and LipBD were overexpressed in soluble form and purified. Those proteins were purified by His-tag affinity chromatography connected in AKTA prime system. The activities of the purified lipases were demonstrated with 1% tributyrin agar plate. After purification, molecular mass was determined with sodium dodecyl sulfate-polyacrylamide gel electrophoresis. LipAD1 showed high activity toward ${\rho}$-nitrophenyl acetate and ${\rho}$-nitrophenyl butyrate, LipAD2 showed high activity toward ${\rho}$-nitrophenyl acetate and ${\rho}$-nitrophenyl myristate, and LipAD3 showed high activity toward ${\rho}$-nitrophenyl acetate, ${\rho}$-nitrophenyl butyrate, and ${\rho}$-nitrophenyl miristate, respectively. Three lipases, LipAD1, LipAD2, and LipAD3, showed optimal reaction at $50^{\circ}C$ using ${\rho}$-nitrophenyl butyrate, as substrate.

Confirmation of Enzymatic Synthesis of 1, 2-Octanediol Galactoside using Mass Spectrometry and NMR Spectroscopy (Mass spectrometry와 NMR Spectroscopy를 이용한 1, 2-Octanediol Galactoside의 효소합성 확인)

  • Lee, Hyang-Yeol;Jin, Hong-Jong;An, Seung Hye;Lee, Hye Won;Jung, Kyung-Hwan
    • Journal of the Korean Applied Science and Technology
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    • v.38 no.3
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    • pp.824-831
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    • 2021
  • 1, 2-Octanediol galactoside (OD-gal) has been synthesized from 1, 2-octanediol (OD), as a safer cosmetic preservative, using recombinant Escherichia coli β-galactosidase (β-gal). To confirm the molecular structure of synthesized OD-gal, mass spectrometry and NMR (1H- and 13C-) spectroscopy of OD-gal were carried out. In the reaction mixture, a sodium adduct ion of OD-gal (m/z=331.1732) was identified using mass spectrometry analysis. In addition, 1H NMR spectrum of OD-gal showed multiple peaks corresponding to the galactosyl group, which is evidence of galactosylation on OD. Downfield proton peaks at δH 4.39 ppm and multiple peaks from δH 3.98~3.55 ppm were indicative of galactosylation on OD. Up field proton peaks at δH 1.52~1.26 ppm and 0.89 ppm showed the presence of CH2 and CH3 protons of OD. 13C NMR spectrum revealed the presence of 24 carbons suggestive of α- and β-anomers of OD-gal. Among 14 carbon peaks from each anomer, the 4 peaks at δC 31.4, 29.0, 22.3, and 13.7 ppm were assigned to be overlapped showing only 24 peaks out of a total of 28 peaks. The mass value from mass spectrometry analysis of OD-gal, and 1H and 13C NMR spectral data were in good agreement with the expecting structure of OD-gal. Finally, we identified a galactose molecule from the hydrolysate of OD-gal using β-gal. We are expecting that through future study it will eventually be able to develop a safe cosmetic preservative.

Improving the Nutritional Value of Tenebrio molitor Larvae by Feeding Them a Soymilk Residue-added Food Source (두유박 첨가 먹이원 급이에 따른 갈색거저리 유충의 영양성 증진 효과)

  • So-Yun Kim;Min Ji Park;Jeong-Hun Song;Sangmin Ji;Gyu-Dong Chang;Sun Young Kim
    • Journal of Life Science
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    • v.34 no.3
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    • pp.191-198
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    • 2024
  • To verify the nutritional value and safety of larvae fed with a soymilk residue-added food source, we compared and analyzed the nutritional components of and harmful substances in Tenebrio molitor larvae reared on wheat bran. Crude protein content based on dry weight was 1.2 times higher in the 10% soymilk residue-fed group (SR) (54.0%) than in the wheat bran-fed group (WB) (43.5%). Dietary fiber also tended to be 1.9 times more in the SR (4.9%) than the WB (2.5%). Among unsaturated fatty acids, the linoleic acid content was found to be 1.1-fold higher in the SR (32.9%) than in the WB (29.0%). Potassium, which was the most abundant among the macro minerals, was 1.1-fold more abundant in the WB (1,074.5 mg/100 g) than in the SR (1,014.0 mg/100 g). Among the micro minerals, zinc content was 1.2-fold higher in the SR (14.5 mg/100 g) than in the WB (11.9 mg/100 g). The results of the analysis of hazardous substances in the WB and SR revealed that the amount of heavy metals met the standards for heavy metals in edible insects, and food poisoning-inducing bacteria such as Escherichia coli and Salmonella spp. were not detected in all groups. These results indicate that mealworm larvae fed with 10% soymilk residue have abundant nutrients and are safe for intake. Thus, food sources with added soymilk residue has the potential to be used as feed ingredients.

Seasonal Variations of Water Quality in the Lower Part of the Nagdong River (낙동강 하류수질의 계절적 변화)

  • KIM Yong-Gwan;SHIM Hye-Kung;CHO Hak-Rae;YOU Sun-Jae
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.17 no.6
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    • pp.511-522
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    • 1984
  • The Nagdong is one of the biggest rivers in Korea, which is very important water source not only for tap water of Pusan city but also for the industrial water. Therefore, authors tried to check the water quality year by year. In this experiment one hundred and twenty water samples collected from August 1983 to July 1984 were analyzed bacteriologically and physiologically. Fifteen sampling stations were established between near Samrangjin and estuary of the river. To evaluate the water quality, temperature, pH, chloride ion, salinity, chemical oxygen demand (COD), electrical conductivity, nutrients, total coliform, fecal coliform, fecal streptococcus, viable cell count and bacterial flora were observed. The variation of water temperature was ranged $-1.5{\sim}29.0^{\circ}C$ (Mean value $13.9{\sim}16.5^{\circ}C$), it in spring was higher as $10{\sim}15^{\circ}C$ about $10^{\circ}C$ than in winter and it in autumm was very stabilized as about $20^{\circ}C$ at each station. The pH variation of the samples was ranged $6.68{\sim}9.15$. The range of concentration of chloride ion and salinity varied $7.4{\sim}l,020.5$ mg/l and $1.05{\sim}33.0\%0$, respectively. Especially, salinity of the 3rd water war was the higher than others as $25.76{\sim}31.58\%0$. COD was ranged $1.45{\sim}14.94$ mg/l and the lower part of the Nagdong River was heavily contaminated by domesitc sewage and waste water from the adjacent factor area. The range of electrical conductivity was $1.360{\times}10^2{\sim}5.650{\times}10^4{\mu}{\mho}/cm$ and that was by far higher the estuary than the upper. Concentration of nutrients were $0.008{\sim}0.040$ mg/l (Mean value $0.019{\sim}0.068$ mg/l) for $NO_2-N,\;0.038{\sim}5.253$ mg/l ($0.351{\sim}2.347$ mg/l) for $NO_3-N,\;0.100{\sim}2.685$ mg/l($0.117{\sim}1.380$ mg/l) for $NH_4-N,\;0.003{\sim}0.084$ mg/l($0.014{\sim}0.065$ mg/l) for $PO_4-P$ and $0.154{\sim}6.123$ mg/l ($1.165{\sim}3.972$ mg/l) for $SiO_2-Si$, respectively. Usually nutrients contents of the water in the upper part(included station 1 to 5) were higher than those of the estuarine area. The bacterial density of the samples ranged 7.3 to 460,000/100 ml for total coliforms, 3.6 to 460,000/100 ml for fecal coliform, $0{\sim}46,000/100ml$ for fecal streptococcus and $<30{\sim}1.2{\times}10^5/ml$ for viable cell count. Composition of coliform was $28\%$ Escherichia coli group, $18\%$ Citrobacter freundii group, $31\%$ Enterobacter aerogenes group and $22\%$ others. Predominant species among the 659 strains isolated from the samples were Pseudomonas spp. ($42\%$), Flavobacterium spp. ($20\%$) and Moraxella spp. ($12\%$).

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Origin of Organic Matter and Geochemical Variation of Upper Quaternary Sediments from the Ulleung Basin (울릉분지 상부 제4기 퇴적물의 유기물 기원 및 지화학적 분포)

  • Kim, Ji-Hoon;Park, Myong-Ho;Ryu, Byong-Jae;Lee, Young-Joo;Oh, Jae-Ho;Cheong, Tae-Jin;Chang, Ho-Wan
    • Economic and Environmental Geology
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    • v.40 no.5
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    • pp.605-622
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    • 2007
  • Elemental, Rock-Eval pyrolysis and isotopic analysis of the core sediments from the northwestern and eastern Ulleung Basin of the East were carried out to identify their geochemical characteristics, spatial and vertical variation and origin of organic matter in Upper Quaternary sediments from the northwestern and eastern Ulleung Basin of the East Sea. TOC, m and TS did not show spatial variation between the sampling locations whereas they showed systematic vertical variation associated with MIS stages related to the sea-level change of the East Sea. It is suggested that these past changes of sea-level influenced the sedimentary depositional environments and/or diagenesis which resulted the patterns observed in this study. Based on the results of TOC/N, TS/TOC, ${\delta}^{13}C_{org}\;and\;{\delta}^{15}N_{org}$ analysis, organic matters in the study area appears to be predominantly originated from the marine algae rather than land plant and deposited under normal marine oxic condition during MIS I and MIS III period, and under euxinic/anoxic condition during MIS II period. TOC/N, ${\delta}^{13}C_{org}\;and\;{\delta}^{15}N_{org}$ have a relatively constant value irrespective of MIS stages, implying that the organic matter source does not change by the sea-level fluctuations. However, the results of Rock-Eval pyrolysis indicates that the organic matter is in immature stage and originated from land-plant (Type III), locating in the immature stage land plant (Type III). Similar differences were reported from other areas such as the Atlantic Ocean, Iberia Abyssal Plain, Mediterranean Sea, suggesting that Rock-Eval method does not exactly reflect the characteristic of immature organic matters. Accordingly, the application of Rock-Eval pyrolysis for delineating the source of immature organic matters should be approached with caution and all other geochemical proxies should be considered altogether at the same time.