• Title/Summary/Keyword: CO I gene

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The Utility of MAGE Gene Detection in Bronchial Washing Fluid for Patients with Peripheral NSCLC (말초 비소세포폐암 환자에서 기관지 세척액 MAGE 유전자 진단의 유용성)

  • Kim, Suhyun;Kim, Hojoong;Kwon, O Jung;Chung, Man Pyo;Suh, Gee Young;Koh, Won-Jung;Ham, Cho Rom;Nam, Hae Seong;Um, Sang-Won;Kwon, Yong Soo;Park, Sung-Hoon
    • Tuberculosis and Respiratory Diseases
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    • v.64 no.1
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    • pp.15-21
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    • 2008
  • Background: The melanoma antigen-encoding (MAGE) genes are known to be expressed in various cancer cells, including non-small cell lung cancer (NSCLC), and are silent in all normal tissues except for the testis. In patients with peripheral NSCLC, bronchial washing fluid can be used to detect the MAGE genes, suggesting a diagnosis of lung cancer. In order to evaluate the diagnostic utility of the MAGE test in patients with peripheral NSCLC, bronchial washing fluid was investigated in patients with peripheral pulmonary nodules, which were invisible as detected by bronchoscopy. Methods: Bronchial washing fluid from 37 patients was used for cytological examinations and MAGE gene detection, using RT-nested-PCR of common A1-A6 mRNA. Results were compared to a final diagnosis of patients as confirmed by pathology. Results: Among the 37 subjects, NSCLC was diagnosed in 21 patients, and benign pulmonary diseases were diagnosed in 16 patients. MAGE mRNA was detected in 10 of 21 (47.6%) NSCLC patients, while conventional cytology examinations were positive for MAGE expression in 2 of 21 (9.5%) cases. MAGE expression was observed in 4 of 16 (25%) benign pulmonary disease patients. Conclusion: The MAGE test of bronchial washing fluid can be used as a sensitive predictor of peripheral NSCLC patients.

Genotypic and Phenotypic Characteristics of Staphylococcus aureus Isolates from Lettuces and Raw Milk (상추와 원유에서 분리한 황색 포도상구균의 유전형 및 표현형 특징)

  • Jung, Hye-Jin;Cho, Joon-Il;Park, Sung-Hee;Ha, Sang-Do;Lee, Kyu-Ho;Kim, Cheol-Ho;Song, Eun-Seop;Chung, Duck-Hwa;Kim, Min-Gon;Kim, Kwang-Yup;Kim, Keun-Sung
    • Korean Journal of Food Science and Technology
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    • v.37 no.1
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    • pp.134-141
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    • 2005
  • To characterize genotypic and phenotypic traits of Staphylococcus aureus isolates (n = 86) from lettuces and raw milk, major virulence-associated genes and antibiotic susceptibility were detected using PCR-based methods and disk diffusion method, respectively. All isolates possessed coagulase gene and showed five polymorphism types [500 bp (2.4%), 580 bp (17.4%), 660 bp (61.6%), 740 bp (17.4%), and 820 bp (1.2%)] due to variable numbers of tandem repeats present within the gene. Two or three different loci of hemolysin gene family were dominant in isolates, 47 of which (55%) possessed combination of hla/hld/hlg-2 genes as the most prevalent types. Among enterotoxin-encoding genes, sea was detected from 32 isolates (37%), sed from 1 isolate (1%), and sea and sed genes were co-detected from 4 isolates (5%), whereas seb, sec, and tsst-1 genes were not detected. All isolates were susceptible to ciprofloxacin, trimethoprim/sulfamethoxazole, oxacillin, and vancomycin, 85 isolates (99%) to penicillin G, 54 isolates (63%) to chloramphenicol, 51 isolates (59%) to erythromycin, and 7 isolates (8%) to clindamycin. Among resistant isolates, seven displayed multiantibiotic-resistance against two different antibiotics.

Phototransduction and Visual Cycle in the Ascidian Tadpole Larva

  • Kusakabe, Takehiro;Nakashima, Yuki;Kusakabe, Rie;Horie, Takeo;Kawakami, Isao;Yoshida, Reiko;Inada, Kyoko;Nakagawa, Masashi;Tsuda, Motoyuki
    • Journal of Photoscience
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    • v.9 no.2
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    • pp.37-40
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    • 2002
  • Ascidians are lower chordates, and their tadpole-like larvae share a basic body plan with vertebrates. To study photoreceptive systems in ascidians, we have isolated and characterized cDNA clones for three opsins, five G protein ${\alpha}$ subunits (G${\alpha}$), catalytic and regulatory subunits of cGMP phosphodiesterase (PDE), and arrestin from the ascidian Ciona intestinalis tadpole larva. Ci-opsin1 and Ci-opsin2 are vertebrate-type opsins, while Ci-opsin3 is a retinal photoisomerase similar to retinochrome and mammalian RGR. Both Ci-opsin1 and arrestin are specifically localized in the photoreceptor cells of the ocellus, whereas Ci -opsin2 is not expressed in the photoreceptors, but is co-localized in another population of neurons in the brain with PDE (Ci-PDE9 and Ci-PDE$\delta$). Ci-opsin3 is present in the entire region of the brain. Though five different cDNAs encoding Ga have been cloned, no transducin-type G protein has been found yet. Interestingly, one of G${\alpha}$i isoform is conspicuously expressed in the entire region of the brain. The Ci-opsin3 gene expression was observed in a broad area of the brain vesicle as well as in the visceral ganglion. Genes encoding ascidian homologs of CRALBP and ${\beta}$-CD, whose function is required for the mammalian visual cycle, are co-expressed with Ci-opsin3 in the brain vesicle and visceral ganglion. Localization of Ci-opsin3, CRALBP, and ${\beta}$-CD in a broad area of the brain suggests that the brain of the ascidian larva has a visual cycle system similar to that of the vertebrate RPE. Based on these data, we discuss the evolution of vertebrate visual systems.

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Cellular Responses to Alcohol in Escherichia coli, Clostridium acetobutylicum, and Saccharomyces cerevisiae (알코올에 대한 Escherichia coli, Clostridium acetobutylicum, Saccharomyces cerevisiae의 반응)

  • Park, Ju-Yong;Hong, Chun-Sang;Han, Ji-Hye;Kang, Hyun-Woo;Chung, Bong-Woo;Choi, Gi-Wook;Min, Ji-Ho
    • Korean Chemical Engineering Research
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    • v.49 no.1
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    • pp.105-108
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    • 2011
  • The increased concern for the security of the oil supply and the negative impact of fossil fuels on the environment, particularly greenhouse gas emissions, has put pressure on society to find renewable fuel alternatives. Compared to the traditional biofuel, ethanol, higher alcohols offer advantage as gasoline substitutes because of their higher energy density and lower hygroscopicity. For this reason, microbial fermentation is known as potential producers for sustainable energy carriers. In this study, bacterial responses including cellular and molecular toxicity were studied in three different microorganisms, such as Escherichia coli, Clostridium acetobutylicum, and Saccharomyces cerevisiae. In this study, it was analyzed specific stress responses caused by ethanol and buthanol using four different stress responsive genes, i.e. fabA, grpE, katG and recA. The expression levels of these genes were quantified by semi-quantitative reverse transcription-PCR. It was found that four genes have shown different responsive patterns when E. coli cultures were under stressful conditions caused by ethanol and buthanol, respectively. Therefore, in this study, the stress responsive effects caused by these alcohols and the extent of each stress response can be analyzed using the expression levels and patterns of different stress responsive genes.

Purification and Characterization of Exo-xylanase from Escherichia coli Cells Harboring the Recombinant Plasmid pMGl (재조합 균주 Escherichia coli가 생산하는 Bacillus stearothermophilus Exo-xylanase의 정제 및 특성)

  • 문애란;최용진
    • Microbiology and Biotechnology Letters
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    • v.20 no.5
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    • pp.574-582
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    • 1992
  • Exo-xylanase encoded by the xylA gene of Bacillus stearothermoPhillus was produced from Escherichia coli ]M109 carrying a recombinant plasmid pMGL Synthesis of the enzyme was observed to be cell-associated, and about 94% of the enzyme synthesized was located in the cytoplasmic region. The maximum production was attained when the E. coli strain was grown at $37^{\circ}C$ for 8 hours on the medium containing 0.5% fructose, 1.0% tryptone, 1.0% sodium chloride, and 0.5% yeast extract. The exo-xylanase was purified to homogeneity using a combination of salting out with ammonium sulfate, DEAE-Sepharose CL-6B ion exchange chromatography, Sephadex G-IOO gel filtration, and Sephadex G-150 gel filtration. The' purified enzyme was most active at pH 6.0 and $45^{\circ}C$. $Ca^{2+}$ and $Co^{2+}$ activated the exo-xylanase activity by about 20% while $Ag^{2+}$, $Fe^{2+}$, $Mg^{2+}$ and $Zn^{2+}$ inhibited the enzyme activity by up to 60%. The $K_m$, value on p-nitrophenyl-$\beta$-D-xylanopyranoside was 2.75 mM. The enzyme had a pI value of 4.7. The estimated molecular weight of the native protein was 200,000 daL SDS-polyacrylamide gel electrophoresis analysis suggested that the native enzyme was a trimer composed of three identical 66,000 da!. polypeptides. The purified enzyme efficiently converted all the xylo-oligosaccharides tested to xylose. It was also confirmed that the enzyme split xylans in an exo-manner even though the degree of hydrolysis was fairly low. The xylanolytic enzyme was, therefore, classified to be one of the few bacterial exo-xylanases lacking transferase activity.

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Evaluation of Optimal Condition for Recombinant Bacterial Ghost Vaccine Production with Four Different Antigens of Streptococcus iniae-enolase, GAPDH, sagA, piaA (연쇄구균증 항원-enolase, GAPDH, sagA, piaA에 대한 재조합 고스트 박테리아 백신의 생산 최적화)

  • Ra, Chae-Hun;Kim, Yeong-Jin;Son, Chang-Woo;Jung, Dae-Young;Kim, Sung-Koo
    • Journal of Life Science
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    • v.19 no.7
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    • pp.845-851
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    • 2009
  • A vector harboring double cassettes; a heterologous gene expression cassette of pHCE-InaN-antigen and a ghost formation cassette of pAPR-cI-E lysis 37 SDM was constructed and introduced to E. coli DH5a. For the production of a bacterial ghost vaccine, bacterial ghosts from E. coli / Streptococcus iniae with four different types of antigens - enolase, GAPDH, sagA and piaA - were produced by the optimization of fermentation parameters such as a glucose concentration of 1 g/l, agitation of 300 rpm and aeration of 1 vvm. Efficiency of ghost bacteria formation was evaluated with cultures of OD$_{600}$=1.0, 2.0 and 3.0. The efficiency of the ghost bacteria formation was 99.54, 99.67, 99.99 and 99.99% with inductions at OD$_{600}$=3.0, 1.0, 2.0 and 1.0 for E. coli/S. iniae antigens enolase, piaA, GAPDH and sagA, respectively. Ghost bacteria as a vaccine was harvested by centrifugation. The antigen protein expressions were analyzed by SDS-PAGE and western blot analysis, and the molecular weights of the enolase, piaA, GAPDH and sagA were 78, 26, 67 and 26 kDa, respectively. The molecular weights of the expressed antigens were consistent with theoretical sizes obtained from the amino acid sequences.

Regulation of the plasminogen activator activity and inflammatory environment via transforming growth factor-beta regulation of sperm in porcine uterine epithelial cells

  • Kim, Su-jin;Cheong, Hee-Tae;Park, Choon-keun
    • Journal of Animal Reproduction and Biotechnology
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    • v.35 no.4
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    • pp.297-306
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    • 2020
  • The aims of the present study were to confirm that regulation of the PA and environment via TGF-β regulation of sperm by Percoll-separated in porcine uterine epithelial cells. And, it was performed to identify the cytokines (TGF-β1, 2 and 3, TGF-β receptor1 and 2; interleukin, IL-6, IL-8) and PA-related genes (urokinase-PA, uPA; tissue-PA, tPA; PA inhibitor, PAI; uPA-receptor, uPAR) by spermatozoa. The experiment used porcine uterus epithelial cells (pUECs) and uterine tissue epithelial cells, Boar sperm were separated by discontinuous Percoll density gradient (45/90%), and tissues were co-incubated with spermatozoa, followed by real-time PCR. PA activity was measured of sperm by discontinuous Percoll density gradient (45/90%) for 24 hours. To measure viability and acrosome damage of sperm double stained propidium iodide (PI) and SYBR-14 or FITC-PNA were used. In results, binding ratio of Percoll-separated sperm was found no differences, but sperms isolated from 90% Percoll layer reduced PA activity (p < 0.05). when co-cultured sperm selected Percoll in porcine uterus tissues epithelial cells, 90% layer sperm increased TGF-β R1, contrastively tPA and PAI-1 in comparison with control (p < 0.05). 45% sperm was decreased the expression of uPA (p < 0.05). TGF-β decreased PA activity in the supernatant collected from pUECs (p < 0.05). Especially, The group including uPA, PAI-1 were induce sperm intact, while it was reduced in sperm damage when compared to control (p < 0.05). Also, there was no significant difference group of tPA and tPA+I in the dead sperm and acrosome damage compared to control. The expression of tPA and PAI showed a common response. Percoll-separated spermatozoa in 90% layer reduced tPA and IL-related gene mRNA expression. Thus, Percoll-sparated sperm in 90% layer show that it can suppress inflammation through increased expression of TGF-β and downregulation of PA and IL in epithelial cells compared to 45% layer Percoll.

Skin Moisturizing Activity of Unripe Apple(Immature Fruit of Malus pumila) in Mice (풋사과 추출물의 피부 보습 효과)

  • Park, Hye Rim;Kim, Jae Kwang;Lee, Jae Kyoung;Choi, Beom Rak;Kim, Jong Dae;Ku, Sae Kwang;Jegal, Kyung Hwan
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.35 no.4
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    • pp.63-74
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    • 2022
  • Objectives : Skin aging is generally characterized by wrinkles, sagging, loss of elasticity roughness, pigmentation and dryness. This changes is caused by reducing the elements constituting the extracellular matrix contributing to the physiological properties of the skin, such as collagen fiber, elastic fiber, and hyaluronic acid. Adequate skin hydration is important to maintain normal skin function and reduce skin aging. The present study is objective to observe skin moisturizing effects of Unripe apple(UA, Immature fruit of Malus pumila Mill) in vivo and its underlying molecular mechanisms. Methods : ICR mice were orally administerd UA(100, 200 and 400mg/kg/day) for 8 weeks, and skin water contents and the expression of transforming growth factor (TGF)-𝛽1, ceramide, hyaluronan and collagen type I(COL1) were measured in dorsal back skin of the mice. Gene expression of hyaluronan synthase(HAS1, HAS2, HAS3), collagen synthase(COL1A1, COL1A2) and TGF-𝛽1 were also determined by realtime RT-PCR. Results : Skin water contents and the expression of TGF-𝛽1, ceramide, COL1 and hyaluronan were significantly increased in UA group(100, 200 and 400mg/kg/day) compared to vehicle control. The mRNA expression of HAS isoform(HAS1, HAS2, HAS3), COL1A1, COL1A2, and TGF-𝛽1 were also significantly increased by UA. Conclusions : UA has skin moisturizing effects and enhancement activities in skin function related components(COL1, hyaluronan, ceramide and TGF-𝛽1). These results suggested that UA can be a developing candidate for developing alternative skin protective agent or functional food ingredient.

Alterations and Co-Occurrence of C-MYC, N-MYC, and L-MYC Expression are Related to Clinical Outcomes in Various Cancers

  • Moonjung Lee;Jaekwon Seok;Subbroto Kumar Saha;Sungha Cho;Yeojin Jeong;Minchan Gil;Aram Kim;Ha Youn Shin;Hojae Bae;Jeong Tae Do;Young Bong Kim;Ssang-Goo Cho
    • International Journal of Stem Cells
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    • v.16 no.2
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    • pp.215-233
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    • 2023
  • Background and Objectives: MYC, also known as an oncogenic reprogramming factor, is a multifunctional transcription factor that maintains induced pluripotent stem cells (iPSCs). Although MYC is frequently upregulated in various cancers and is correlated with a poor prognosis, MYC is downregulated and correlated with a good prognosis in lung adenocarcinoma. MYC and two other MYC family genes, MYCN and MYCL, have similar structures and could contribute to tumorigenic conversion both in vitro and in vivo. Methods and Results: We systematically investigated whether MYC family genes act as prognostic factors in various human cancers. We first evaluated alterations in the expression of MYC family genes in various cancers using the Oncomine and The Cancer Genome Atlas (TCGA) database and their mutation and copy number alterations using the TCGA database with cBioPortal. Then, we investigated the association between the expression of MYC family genes and the prognosis of cancer patients using various prognosis databases. Multivariate analysis also confirmed that co-expression of MYC/MYCL/MYCN was significantly associated with the prognosis of lung, gastric, liver, and breast cancers. Conclusions: Taken together, our results demonstrate that the MYC family can function not only as an oncogene but also as a tumor suppressor gene in various cancers, which could be used to develop a novel approach to cancer treatment.

Methane Production and T-RFLP Patterns of Methanogenic Bacteria Dependent on Agricultural Methods (농법에 따른 메탄생성과 메탄생성 세균의 T-RFLP 패턴)

  • Kim, Hun-Soo;Cho, Ju-Sik;Park, Kyeong-Ryang
    • Korean Journal of Microbiology
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    • v.45 no.1
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    • pp.17-25
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    • 2009
  • We studied soil components, methane production, the number of methanogens, and T-RFLP patterns dependent on agricultural methods with the change of seasons. There is no regular increase or decrease tendency of the most soil components followed by sampling period. And the water content in soil was higher in October than May. Also a lot of methanogens existed in soil, and acetotrophs were relatively of smaller number than hydogenotrophs and formate utilizing methanogens using MPN (most probable number) enumeration. In the experiment using the formate, it was used from the first week, and only a minute amount was detecte after four weeks. However in the acetate, it was increased until the third week, and after that was consumed. And there was higher methane production for all soil samples which administered with the hydrogen spike. The activity of methanogens was higher in the organic and low-agrichemical agricultural method samples, and the organic agricultural method had high methanogen activity among the other samples. A result of T-RFLP pattern of mcrA gene digested with Sau96I, methanogen community have a little relation with agricultural methods and seasons. This results also agreed to no critical difference the soil components dependent on agricultural methods, but some analytical data have a positive relationship with a agricultural methods. Therefor we could concluded that the comparison study of community for soil bacteria sufficiently could be useful for the microbiological indicator.