• Title/Summary/Keyword: Isolated pattern

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Interferon-gamma susceptibility of HL-60 cells, mononuclear cells of umbilical cord blood and bone marrow (HL-60 세포주, 제대혈 및 골수 단핵구 세포의 interferon-gamma에 대한 감수성에 관한 연구)

  • Cheong, Hee Jeong;Hong, Dae Sik;Kim, Sook Ja;Cheong, Jae Hwa;Lee, Joo Young;Lee, Nam Su;Park, Sung Kyu;Won, Jong Ho;Park, Hee Sook;Kim, Sung Il
    • IMMUNE NETWORK
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    • v.1 no.3
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    • pp.230-235
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    • 2001
  • Background: Finding of the regulation of various gene expression by cytokine including $IFN-{\gamma}$ in hematopoietic stem cell will light up the understanding of pathogenesis of aplastic anemia in various aspects. To study on aplastic anemia, however, we have to circumvent the difficulty of directly obtaining bone marrow stem cells from the patient. Therefore, we tried to find out a cell can replace the bone marrow stem cells for study on cell signaling pathway and regulation of gene expression by $IFN-{\gamma}$. Materials and Methods: HL-60 cells, of 20 ng/mL of $IFN-{\gamma}$. Total RNA was isolated from the cells and RT-PCR of the indoleamine 2,3-dioxygenase (IDO), $IFN-{\gamma}$, TNF-${\alpha}$, $MIP-1{\alpha}$, and $TGF-{\beta}2$ was carried out for the estimation of the gene expression. Results: $IFN-{\gamma}$ induced IDO gene expression of mononuclear cells from umbilical cord blood showed similar pattern as compared to that of bone marrow. Whether $INF-{\gamma}$ was treated or not, $TNF-{\alpha}$ was expressed in both mononuclear cells from umbilical cord blood and bone marrow. However, HL-60 cells showed different expression patterns. HL-60 cells would express neither IDO nor $TNF-{\alpha}$ even under the culture with 20ng/mL of $IFN-{\gamma}$. Conclusion: Our results showed bone marrow can be replaced with mononuclear cells from umbilical cord blood in the study on the relation between aplastic anemia and $IFN-{\gamma}$ including $IFN-{\gamma}$ cell signaling pathway.

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Human Cord Serum as a Fetal Bovine Serum Substitute for the Culture of Human Amnion-Derived Stem Cells (인간의 양막유래 줄기세포의 체외 배양 시 소태아혈청 대체제로서의 인간제대혈청)

  • Kim, Jin-Young;Park, Se-A;Kang, Hyun-Mi;Kim, Eun-Su;Kim, Hae-Kwon
    • Development and Reproduction
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    • v.11 no.2
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    • pp.85-96
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    • 2007
  • Mesenchymal stem cells (MSC) are promising candidates for cell-based therapies. One major obstacle for their clinical use is the unsafety of fetal bovine serum (FBS), which is a crucial part of all media currently used for the culture of MSC. We investigated the effect of human cord serum (HCS) on the growth response, mRNA and protein expressions of human amnion-derived stem cells (HAM). HAM were isolated from the amnion after a Caesarean section and cultured in DMEM supplemented with 10% FBS, 5% HCS or 10% HCS. During culture, their biological characteristics at earlier and later passages were analyzed using RT-PCR and immunocytochemistry. Regardless of serum sources, HAM showed the prominent expression of Oct-4, Rex-1, SCF, FGF-5, BMP-4, nestin, GATA-4, NCAM and HLA ABC genes. The expression profile was observed even at later passages. Similarly, HAM cultured in either FBS or HCS exhibited the distinct protein expression of collagen I, II, III and XII, fibronectin, $\alpha$-smooth muscle actin, vimentin, CK18, CD54, FSP, TRA-1-60, SSEA-3, -4 and HLA ABC. However, desmin expression was only observed in HAM cultured in medium supplemented with FBS and vWF expression was only found in HAM cultured in medium supplemented with HCS. Overall pattern of gene and protein expression of HAM was typical of known adult stem cells such as bone marrow-derived MSC. In conclusion, HCS could be as effective as FBS for the culture of HAM.

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Geological Structures and Evolution of the Tertiary Chŏngja Basin, Southeastern Margin of the Korean Peninsula (울산군 강동면 제 3기 정자분지(亭子盆地)의 지질구조와 분지발달)

  • Son, Moon;Kim, In-Soo
    • Economic and Environmental Geology
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    • v.27 no.1
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    • pp.65-80
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    • 1994
  • The Tertiary $Ch{\check{o}}ngja$ basin is located in the southeastern coastal area of the Korean Peninsula. It is a lozenge shaped fault-bounded basin with circa $5{\times}5km$ areal extent, isolated from other Tertiary basins by the Cretaceous Ulsan Formation in-between. The northwestern boundary of the basin is a domino/listric type normal fault trending $N30^{\circ}E$, whereas its southwestern boundary is a dextral strike-slip fault (trending $N20^{\circ}W$) with a lateral offset of more than 1 km. The basin is bounded by the East Sea on the eastern margin. Basin-fills consist of extrusive volcanic rock (Tangsa Andesites) of Early Miocene (16~22 Ma in radiometric age), unconsolidated fluviatile conglomerate (Kangdong Formation) and shallow brackish-water sandstone ($Sinhy{\check{o}}n$ Formation). The latter yields abundant Vicarya-Anadara molluscan fossils of early Middle Miocene age. The Tertiary strata become younger toward the northwestern boundary-fault of the basin, showing a zonal distribution pattern parallel to the fault: the younger sedimentary formations occupy a narrow zone of 2 km width along the northwestern boundary-fault, whereas the older Tangsa Andesites underlie them unconformably in the eastern and southeastern portions of the basin. The strata in the basin, including the Tangsa Andesites, are tilted (about $20^{\circ}$) toward the northwestern boundary-fault Sedimentary strata thicken toward the boundary-fault, forming a wedge shaped half-graben structure. A number of small-scale syndepositional normal growth faults and graben structures are observed in the sedimentary strata. These extensional structures have the same trend as the normal northwestern boundary-fault which we interpret as a pull-apart detachment fault. These characteristics imply persistent extension during the basin evolution, caused by a NW-SE directed tensional force. The $Ch{\check{o}}ngja$ basin is, thus, a kind of syndepositional tectonic basin evolved in a strike-slip (pull-apart) regime. The latter was caused by a dextral simple shear associated with the NNW-SSE opening of the East Sea. In view of the fact that the normal growth faults do not cut through the uppermost portion of the youngest $Sinhy{\check{o}}n$ Formation, it is inferred that the tensional force came to be inactive in the early Middle Miocene. This is coincident in timing with the termination of the East Sea opening (15 Ma).

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Serological Distribution and Properties of Antibiotic Resistance of Escherichia coli from Patients with Diarrhea (설사환자로부터 분리한 대장균의 혈청형 분포 및 항생제 내성유형)

  • 차인호;진성현;박은희;박성아;조현철;이영숙;정석훈;이영길;이상훈
    • Journal of Life Science
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    • v.10 no.3
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    • pp.262-272
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    • 2000
  • As a part of investigation for basic epidemiology of diarrheogenic disease, we attempted isolation of Escherichia coli from patients with diarrhea. Seven hundred and twenty-one strains of E. coli were isolated from 1,239 patients with diarrhea. Seasonal distribution of patient with diarrhea was shown the most high at August (18.2%). Age group distribution of patient was shown the most high at children (54.6%, 2 to 10 years old). The serotypes of 721 E. coli isolates were in order of serotype O44 (16.8%), O153 (8.6%), O1 (7.5%), O166(5.7%), O8 and O86a (4.7%), and O125 (4.6%). The supernates cultured 36 strains among 721 E. coli isolates were indicated cytotoxicity against monolayered Vero cells. All of the isolates were susceptible to amikacin. The isolates were resistant in order of novobiocin (99.0%), moxalactam (97.1%), carbenicillin (96.1%), tetracycline (90.4%), ampicillin (85.9%), gentamicin (84.0%), streptomycin (78.4%), cephalothin (46.6%) and polymyxin B (4.2%). In the antibiotic resistant patterns, 125 kinds of multiple resistance patterns of E. coli isolates were detected. The highest resistant pattern was ampicillin-carbenicillin-chloramphenicol-cephalothin-erythro-mycin- gentamicin-moxalactam-novobiocin-penicillin G-streptomycin-tobramycin-tetracycline-tri methoprim type (24.3%).

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Isolation and Partial Characterization of Isoflavone Transforming Lactobacillus plantarum YS712 for Potential Probiotic Use (Isoflavone 비배당화 및 항산화 활성을 지닌 Lactobacillus plantarum YS712의 선발)

  • Cho, Yoon-Hee;Imm, Jee-Young;Kim, Hwa-Young;Hong, Seong-Gil;Hwang, Sung-Joo;Park, Dong-Jun;Oh, Se-Jong
    • Food Science of Animal Resources
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    • v.29 no.5
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    • pp.640-646
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    • 2009
  • Lactic acid bacteria (LAB) are typical probiotic microbes that are used in various industries including fermented foods, feed additives, and pharmaceuticals. The purpose of this study was to compare the ability of isoflavone biotransformation and antioxidative activity of 17 LAB. Six strains including the Lactobacillus species exhibited a 100% hydrolysis rate for daidzein during fermentation. The content of total genistein in soymilk fermented with these strains was $872-943\;{\mu}g/g$. The DPPH (1, 1-diphenyl-2-picrylhydrazyl) radical scavenging ability of the LAB was widely variable and ranged from 23-78%. A selected strain was isolated from kimchi and the strain was identified as Lactobacillus plantarum ssp. through the API carbohydrate fermentation pattern and 16S rDNA profile. The strain exhibited excellent acid tolerance in an artificial gastric solution. L. plantarum YS712 showed high $\beta$-glucosidase activity in fermentation. The concentration of genistein and daidzein in soymilk fermented with L. plantarum YS712 increased from 3.64 to $917.3\;{\mu}g/g$ and from 58.18 to $1062.17\;{\mu}g/g$, respectively. These results demonstrate the potential of L. plantarum YS712 as a probiotic culture that can be utilized in the manufacturing of fermentation foods and dietary supplements.

Characteristics and Action Pattern of Alkaline Protease from Streptomyces gviseus HC-1141 (Streptomyces griseus HC-1141이 생성하는 Alkaline Protease의 특성 및 작용양상)

  • Choi, Cheong;Chung, Yung-Gun;Sung, Sam-Kyung;Choi, Kwang-Soo;Lee, Jae-Sung;Cho, Young-Je;Chun, Sung-Sook
    • Microbiology and Biotechnology Letters
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    • v.20 no.3
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    • pp.295-301
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    • 1992
  • An alkaline protease producing microorganism was isolated from soil and identified as Streptomyces griseus HC-1141. The optimum pH and temperature for the purified enzyme activity were 8.0 and $60^{\circ}C$, respectively. The enzyme was relatively stable in the pH range of 7.0-9.0 and at the temperature below $60^{\circ}C$. The activity of purified enzyme was inhibited by $Hg^{2+}$, $Cu^{2+}$, $Zn^{2+}$, $Ba^{2+}$ and $Fe^{2+}$, whereas activated by $Mn^{2+}$ and $Ca^{2+}$. $\varepsilon$-Amino caproic acid, 2,4-dinitrophenol and iodine did not show inhibitory effect on the activity of alkaline protease, but p-chloromercuribenzoic acid, ethylendiaminetetraacetic acid showed inhibitory effect on the enzyme activity. These result suggested that the protease was metalloenzyme, and require a reactive SH group for the activity. The reaction of this enzyme follows typical Michaelis-Menten kinetics with the $K_m$ value of $2.229{\times}10^{-4}$M and the $V_{max}$ of $46.08 {\mu}$g/min for casein. The activation energy for the alkaline protease calculated by Arrhenius equation was 3.643 kcal/mol. This enzyme hydrolyzed casein more rapidly than the hemoglobin and egg albumin.

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Detection of DNA from Dermatophytes by Polymerase Chain Reaction (Polymerase chain reaction에 의한 동물 유래 피부사상균 DNA의 검출)

  • Kim, Young-Wook;Yeo, Sang-Geon;Choi, Woo-Pil
    • Korean Journal of Veterinary Research
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    • v.42 no.3
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    • pp.363-370
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    • 2002
  • For the development of diagnostic polymerase chain reaction (PCR) to fungal infection by dermatophytes Trichophyton and Microsporum, detection of the fungal DNA by PCR and analysis of the DNA pattern were undertaken in the present study. A total of 15 strains were tested and those consisted of 3 reference strains and 12 isolates such as: reference strains of T mentagrophytes (downy type, ATCC 9533), T rubrum (IFO 6204) and M gypseum (ATCC 9083), and each isolate of T mentogrophytes (powdery type), T mentagrophytes (granular type), T mentogrophytes (purple-red type), T rubrum, T raubitschekii, T tonsurans, T equinum, T ajelloi, T verrucosum, M cookei, M nanum and M gypseum. The DNA were purely isolated from all strains of Trichophyton spp. and Microsporum spp. by a simple method partly consisted of disruption of fungal cells by lyophilization and grinding and extraction of fungal DNA without phenol treatment which is a routine procedure in DNA isolation. For the detection of fungal DNAs, optimal condition of PCR was determined as preheating once at $94^{\circ}C$ for 5 min, 35 cycles of denaturation at $94^{\circ}C$ for 1 min, annealing at $38^{\circ}C$ for 1 min and polymerization at $72^{\circ}C$ for 2 min, and 1 cycle of final extension at $72^{\circ}C$ for 5 min. In PCR using arbitrary primers AP-1 (5' ACCCGACCTG3') and AP-2 (5' ACGGGCCAGT3'), DNAs in various numbers and sizes were detected from different species of Trichophyton and Microsporum, while DNAs in similar size were also detected in all strains of Trichophyton spp. and Microsporum spp. There were unique DNAs observed from certain dermatophytes by AP-1 such as 1,900 bases in T rubrum, 950 and 1,100 bases in T raubitscheldi, 2,100 bases in T equinum, 400 bases in T verrucosum and 1,150 bases in M gypseum. The unique DNAs were also observed by AP-2 such as 1,200 bases in T ajelloi, 250 bases in T verrucosum, 1,150 bases in M cookei and 2,000 bases in M nanum. The results indicated that PCR can detect a specific DNA from certain Trychophyton and Microsporum spp, which can be the information for further development of diagoomc PCR to dennatophytes.

Bacterial Distribution and Antimicrobial Resistance Pattern of PC Room(Internet Cafe) (서울 시내 PC방(Internet Cafe)의 세균분포와 항생제 내성 양상)

  • Yun Ji-Hee;Back Han-Joo;Jin Han-Joo;Son Ye-Won;Kwak Sun-Young;Yang Hwan-Jin;Hong Eun-Kyung;Choi Sung-Suk;Ha Nam-Joo
    • Korean Journal of Microbiology
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    • v.41 no.3
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    • pp.183-187
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    • 2005
  • This study was performed to investigate the hygiene condition of PC room(internet cafe) in Seoul Korea. From July 2004 to December, 34 samples were collected, there's an average of $9.0{\times}10^4$ CFU/ml on keyboards, $2{\times}10$ CFU/ml on mouse and $5{\times}10^3$ CFU/ml on door konbs toilets, suggesting that keyboards and mouse are more contaminated than toilet door knobs. Seven antimicrobial resistant strains were isolated from PC Rooms. Two isolates were resistant to methicillin and erythromycin, while five isolates were resistant to gentamicin, ampicllin, cefotaxim, and chloramphenicol. By identification, these strains were identified as Staphylococcus aureus (2 strains). Actinobacillus ureae (4 strains) and Pasteurella multocida (1 strain), respectively. Pasteurella multocida and Actinobacillus ureae are potentially pathogenic bacteria. Actinobacillus ureae, formerly, known as Pasteurella ureae, is an uncommon of the upper respiratory tract in humans. Pasteurella multocida is a part of the normal flora in the nasopharynx of many domestic animals. We concluded that Staphylococcus aureus is highly resistant to erythromycin and methicillin over $100\;{\mu}g/ml$, while Pasteurella multocida and Actinobacillus ureae is highly resistant to gentamicin, ampicillinover over $100\;{\mu}g/ml$.

Distribution Pattern of Soil Actinomycetes on the Seasonal Change (계절에 따른 토양 방선균의 속 다양성 분포)

  • Park, Dong-Jin;Lee, Sang-Hwa;Park, Sang Ho;Kim, Chang-Jin
    • Korean Journal of Microbiology
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    • v.34 no.3
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    • pp.149-153
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    • 1998
  • From soils seasonally collected at two depths (0~2 cm, $50{\pm}1cm$) of forest, field, grass land, or paddy field, distinct strains of actinomycetes were isolated and identified to the genus level. The genus-diversity of soil actinomycetes was revealed to be considerably different by seasonal change. It was also affected by soil depths, soil types, or actinomycete groups. At the soil depth of 0~2 cm, the seasonal distribution fluctuation (%) of streptomycete strains was higher in grass land (41%), field (39%) soil than paddy field (18%), or forest (18%), whereas that of streptomycete strains at the soil depth of $50{\pm}1cm$ was high in order of paddy field (36%), field (28%), grass land (26%), and forest (16%). On the other hand, the seasonal distribution fluctuation ratio of rare actinomycete strains at the soil depth of 0~2 cm was above 45% except for paddy field (26%). At the soil depth of $50{\pm}1cm$, the seasonal distribution of rare actinomycete strains exhibited high fluctuation (%) in order of forest (79%), paddy field (36%), field (24%), and grass land (10%).

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Electrophoretic Karyotypes of Fusarium oxysporum f. sp. lycopersici (Fusarium oxysporum f. sp. lycopersici의 Electrophoretic Karyotype)

  • Kim, Young-Tae;Kim, Hong-Gi
    • The Korean Journal of Mycology
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    • v.27 no.2 s.89
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    • pp.112-118
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    • 1999
  • Strains of Fusarium oxysporum f. sp. lycopersici isolated from Korea, Japan and U.S.A. were used for electrophoretic karyotype (EK) analysis. Chromosome separations on FastLane agarose gels (FMC BioProducts, Rockland, ME), called pulsed field gel electrophoresis (PFGE), were performed by CHEF-DRII apparatus (Bio-Rad Laboratories, Melville, NY) using TAE as a running buffer. To obtain optimal condition for separation of chromosome sized DNAs, variable running conditions such as field strengths, swithching intervals, and running time were applied in CHEF gel electrophoresis. We were able to resolve 9 to 11 chromosome sized DNAs ranging in size from 0.76 to 6.41 Mb in isolates from Korea and estimate that the total genome size was ranging from 35.29 to 38.92 Mb. Distinct differences in length range and genome size exist among isolates from different countries. Isolates from Japan and U.S.A. were resolved 9 to 11 chromosome sized DNAs ranging in size from 1.24 to 6.85 Mb and estimated that the total genome size was ranging from 35.32 to 43.87 Mb. Isolates from variable provinces in Korea had the same or similar chromosomal polymorphism and showed different chromosomal DNA patterns compared to isolates from the other countries.

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