• 제목/요약/키워드: M2 gene

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시료 중 잔류 항생제 분석 방법: I. 고상 시료 전처리 방법 (Determination of Antibiotic Residues: I. Extraction and Clean-up Methods for Solid Samples_A Review)

  • 김찬식;류홍덕;정유진;김용석;류덕희
    • 한국물환경학회지
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    • 제32권6호
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    • pp.600-627
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    • 2016
  • Korea is one of the countries with a large veterinary antibiotics market, although antimicrobial resistance in bacteria is becoming a serious issue in many countries. The Korean government started to take interest in estimating the effects of livestock manure on rivers and agricultural soils and in monitoring of heavy metals, organic pollutants and antibiotics in the ambient water and soil. In this paper, pre-treatment methods to separate the selected antibiotics from solid samples were reviewed. It is essential to select an efficient and appropriate procedure for pre-treatment due to the high proportion of proteins and organics in biosolid samples. Pre-treatment consists of extraction followed by clean-up. Initially, homogenized samples were extracted by sonication, mechanical agitation or pressurized liquid extraction with methanol/acetonitrile/water mixture under acidic/basic conditions depending on the compound. However, aminoglycosides and colistin were extracted with 5% trichloroacetic acid and HCl, respectively. Since the ${\beta}-lactams$ are easily decomposed in acidic and basic conditions, they were extracted in neutral pH. Filtration with a membrane (pore size, $0.2{\mu}m$) or solid phase extraction with HLB and methanol, as eluents, was normally applied for the clean-up. At least, three different pre-treatment procedures should be adopted to screen all the selected antibiotics in solid samples.

Aldose Reductase Inhibitor Fidarestat as a Promising Drug Targeting Autophagy in Colorectal Carcinoma: a Pilot Study

  • Pandey, Saumya
    • Asian Pacific Journal of Cancer Prevention
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    • 제16권12호
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    • pp.4981-4985
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    • 2015
  • Background: Colorectal cancer (CRC) is a leading cause of morbidity and mortality worldwide. Targeting autophagic cell death is emerging as a novel strategy in cancer chemotherapy. Aldose reductase (AR) catalyzes the rate limiting step of the polyol pathway of glucose metabolism; besides reducing glucose to sorbitol, AR reduces lipid peroxidation-derived aldehydes and their glutathione conjugates. A complex interplay between autophagic cell death and/or survival may in turn govern tumor metastasis. This exploratory study aimed to investigate the potential role of AR inhibition using a novel inhibitor Fidarestat in the regulation of autophagy in CRC cells. Materials and Methods: For glucose depletion (GD), HT-29 and SW480 CRC cells were rinsed with glucose-free RPMI-1640, followed by incubation in GD medium +/- Fidarestat ($10{\mu}M$). Proteins were extracted by a RIPA-method followed by Western blotting ($35-50{\mu}g$ of protein; n=3). Results: Autophagic regulatory markers, primarily, microtubule associated protein light chain (LC) 3, autophagy-related gene (ATG) 5, ATG 7 and Beclin-1 were expressed in CRC cells; glyceraldehyde-3 phosphate dehydrogenase (GAPDH) was used as an internal reference. LC3 II (14 kDa) expression was relatively high compared to LC3A/B I levels in both CRC cell lines, suggesting occurrence of autophagy. Expression of non-autophagic markers, high mobility group box (HMG)-1 and Bcl-2, was comparatively low. Conclusions: GD +/- ARI induced autophagy in HT-29 and SW-480 cells, thereby implicating Fidarestat as a promising therapeutic agent for colorectal cancer; future studies with more potent ARIs are warranted to fully dissect the molecular regulatory networks for autophagy in colorectal carcinoma.

7, 12-dimethylbenz[a]anthracene(DMBA)로 유발된 햄스터 협낭암에서 chlorophylln의 암예방효과에 관한 실험적 연구 (AN EXPERIMENTAL STUDY ON THE CHEMOPREVENTIVE EFFECT OF CHLOROPHYLLIN IN HAMSTER CHEEK POUCH TUMOR INDUCED BY 7, 12-DIMETHYLBENZ[A]ANTHRACENE)

  • 윤규호
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • 제26권2호
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    • pp.137-145
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    • 2000
  • Carcinogenesis is a multi-stage process that generally consists of at least three steps; initiation, promotion, and progression. If one of these carcinogenic steps were suppressed or delayed, the cancer could be prevented. Cancer chemoprevention is defined to be inhibition or reversal of the carcinogenic process by the specific chemical agents and is a novel approach to cancer management alternative to conventional chemotherapy. Chlorophylln(CHL), a water-soluble derivative of chlorophyll, containing sodium and copper, has been known to be strong antimutagen in several test systems, but its mechanism of antimutagenic action is unknown. In the present experiment, the possibility of CHL as chemopreventive drugs on 7,12-dimethylbenz[a]anthracene(DMBA)-induced hamster buccal pouch carcinogenesis was investigated by mutagenicity test, carcinogenicity test, and frequency or spectrum of H-ras mutations in the both of DMBA-induced and chlorophylln-pretreated-DMBA induced tumor by polymerase chain reaction and non-isotopic restriction fragment length polymorphism. The treatment of CHL reduced the yields and multiplicity of the 0.5% DMBA-induced tumor, 86% to 62.5% and $3.7{\pm}0.6$ to $1.4{\pm}0.3$, respectively. The occurrence of histidine revertant by $20{\mu}mole$ DMBA was inhibited 25.6 to 81.7% by 1 to $5{\mu}M$ CHL in a dose-dependent manner. The mutation rates of H-ras gene in DMBA-induced and CHL-pretreated-DMBA induced tumor were 96%, 94% of which the most mutations were in codon 12/13. These results suggest that CHL inhibits the carcinogenic action of DMBA by the formation of complex between CHL and DMBA or the inhibition of the activation of DMBA in vivo. But CHL did not affect the mutation rates or its spectrum in already formed tumor.

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Effects of polysaccharides derived from Orostachys japonicus on induction of cell cycle arrest and apoptotic cell death in human colon cancer cells

  • Ryu, Deok-Seon;Baek, Geum-Ok;Kim, Eun-Young;Kim, Ki-Hoon;Lee, Dong-Seok
    • BMB Reports
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    • 제43권11호
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    • pp.750-755
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    • 2010
  • Crude Orostachys japonicus polysaccharide extract (OJP) was prepared by hot steam extraction. Polysaccharides (OJPI) were separated from OJP by gel filtration chromatography and phenol-sulfuric acid assay. The average molecular weight of the OJPI was 30-50 kDa. The anti-proliferative effect of OJPI on HT-29 human colon cancer cells was investigated via morphology study, cell viability assay, apoptosis assay, cell cycle analysis, and cDNA microarray. OJPI inhibited proliferation and growth of HT29 cells and also stimulated apoptosis in a dose- and time-dependent manner. In cell cycle analysis, treatment with OJPI resulted in a marked increase of cells in the G0 (sub G1) and G2/M phases. To screen for genes involved in the induction of cell cycle arrest and apoptosis, the gene expression profiles of HT-29 cells treated with OJPI were examined by cDNA microarray, revealing that a number of genes were up- or down-regulated by OJPI. Whereas several genes involved in anti-apoptosis, cell proliferation and growth, and cell cycle regulation were down-regulated, expression levels of several genes involved in apoptosis, tumor suppression, and other signal transduction events were up-regulated. These results suggest that OJPI inhibits the growth of HT-29 human colon cancer cells by various apoptosis-aiding activities as well as apoptosis itself. Therefore, OJPI deserve further development as an effective agent exhibiting anticancer activity.

Optimized Medium Improves Expression and Secretion of Extremely Thermostable Bacterial Xylanase, XynB, in Kluyveromyces lactis

  • Yin, Tie;Miao, Li-Li;Guan, Fei-Fei;Wang, Gui-Li;Peng, Qing;Li, Bing-Xue;Guan, Guo-Hua;Li, Ying
    • Journal of Microbiology and Biotechnology
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    • 제20권11호
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    • pp.1471-1480
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    • 2010
  • An extremely thermostable xylanase gene, xynB, from the hyperthermophilic bacterium Thermotoga maritima MSB8 was successful expressed in Kluyveromyces lactis. The response surface methodology (RSM) was also applied to optimize the medium components for the production of XynB secreted by the recombinant K. lactis. The secretion level (102 mg/l) and enzyme activity (49 U/ml) of XynB in the optimized medium (yeast extract, lactose, and urea; YLU) were much higher than those (56 mg/l, 16 U/ml) in the original medium (yeast extract, lactose, and peptone; YLP). The secretory efficiency of mature XynB was also improved when using the YLU medium. When the mRNA levels of 13 characterized secretion-related genes in the K. lactis cultured in YLP and YLU were detected using a semiquantitative RT-PCR method, the unfolded protein response (UPR)-related genes, including ero1, hac1, and kar2, were found to be up-regulated in the K. lactis cultured in YLU. Therefore, the nutrient ingredients, especially the nitrogen source, were shown to have a significant influence on the XynB secretory efficiency of the host K. lactis.

New HDAC inhibitor, IN2001 induces apoptosis/cell cycle arrest in human breast cancer cells

  • Euno, Joung-Ki;Nan, Min-Kyung;Jung, Cho-Min;Young, An-Jin;Kim, -Dae-Ki;Yhong, Sheen-Yhun
    • 한국환경독성학회:학술대회논문집
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    • 한국환경독성학회 2003년도 추계국제학술대회
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    • pp.180-180
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    • 2003
  • The acetylation of histone is one of the mechanisms involved in the regulation of gene expression and is tightly controlled by two core enzymes, histone acetyltransferase (HAT) and deacetylase (HDAC). There are several reports that imbalance of HAT and HDAC activity is associated with abnormal behavior of the cells in morphology, cell cycle, differentiation, and carcinogenesis. Recently, an increasing number of structurally diverse HDAC inhibitors have been identified that inhibit proliferation and induce differentiation and/or apoptosis of tumor cells in vivo and in vitro. In this study, we have investigated the effects of novel HDAC inhibitors, IN2001 on ER positive and ER negative human breast cancer cell lines. The growth inhibition, cell cycle arrest and apoptosis of cells by HDAC inhibitors were determined using SRB assay, DNA fragmentation, and flow cytometry. We found that IN 2001 as well as Trichostatin A inhibited cell growth dose-dependently in both ER Positive and ER negative human breast cancer cell lines. The growth inhibition with HDAC inhibitors was associated with profound morphological change. The result of cell cycle analysis after 24 h exposure of IN2001 showed G2-M cell cycle arrest in MCF-7 cell and apoptosis in T47B and MDA-MB-231 cell. In summary, IN2001 has antiproliferative effect on human breast cancer cells regardless of the expression of estrogen receptor. These findings heights the possibility of developing HDAC inhibitors as potential anticancer therapeutic agents for the treatment of breast cancer.

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해수 및 시판 수산물에서 분리한 장염비브리오균(Vibrio parahaemolyticus)의 항균제 내성 및 최소발육억제농도의 규명 (Antimicrobial Resistance and Minimum Inhibitory Concentrations of Vibrio parahaemolyticus Strains Isolated from Seawater and Commercial Fisheries)

  • 조의동;김희대;박권삼
    • 한국수산과학회지
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    • 제52권6호
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    • pp.587-595
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    • 2019
  • Eighty-three Vibrio parahaemolyticus isolates from surface seawater in Gomso Bay on the west coast of Korea, and commercial fisheries from Gunsan fisheries center were analyzed for the presence of virulence genes and susceptibility to 30 different antimicrobials. All 83 isolates were examined for the presence of two virulence genes (tdh or trh) using polymerase chain reaction; however, neither gene was found in any of the isolates. A disk diffusion susceptibility test, showed that all of the strains studied were resistant to clindamycin, oxacillin, ticarcillin, and vancomycin, and also revealed varying levels of resistance to ampicillin (98.8%), penicillin G (95.2%), streptomycin (20.5%), cefoxitin (14.5%), amikacin (6.0%), cephalothin (4.8%), and erythromycin (3.6%). However, all of the strains were susceptible to 19 other antimicrobial agents, including cefepime, cefotaxime, chloramphenicol, gentamycin, nalidixic acid, sulfamethoxazole/trimethoprim, and trimethoprim. All 83 isolates (100%) were resistant to five or more classes of antimicrobials, and two strains exhibited resistance to ten antimicrobial agents. The average minimum inhibitory concentrations against V. parahaemolyticus of clindamycin, oxacillin, ticarcillin, and vancomycin were 55.9, 98.3, 499.3, and 44.3 ㎍/mL, respectively. These results provide new insight into the necessity for seawater sanitation in Gomso Bay and commercial fisheries, and provide evidence to help reduce the risk of contamination by antimicrobial-resistant bacteria.

TGF-β Signaling and miRNAs Targeting for BMP7 in the Spleen of Two Necrotic Enteritis-Afflicted Chicken Lines

  • Truong, Anh Duc;Hong, Yeojin;Lee, Janggeun;Lee, Kyungbaek;Lillehoj, Hyun S.;Hong, Yeong Ho
    • 한국가금학회지
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    • 제44권3호
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    • pp.211-223
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    • 2017
  • Transforming growth factor beta ($TGF-{\beta}$) signaling pathways are involved in the regulation of proliferation, differentiation, immunity, survival, and apoptosis of many cells. The aim of this study was to investigate the differential expression of $TGF-{\beta}$-related genes, and their interactions and regulators in the spleen of two genetically disparate chicken lines (Marek's disease resistant line 6.3 and Marek's disease-susceptible line 7.2) induced with necrotic enteritis (NE) by Eimeria maxima and Clostridium perfringens infection. By using high-throughput RNA-sequencing, we investigated 76 $TGF-{\beta}$-related genes that were significantly and differentially expressed in the spleens of the chickens. Approximately 20 $TGF-{\beta}$ pathway genes were further verified by qRT-PCR, and the results were consistent with our RNA sequencing data. All 76 identified genes were analyzed through Gene Ontology and mapped onto the KEGG chicken $TGF-{\beta}$ pathway. Our results demonstrated that several key genes, including $TGF-{\beta}$1-3, bone morphogenetic proteins (BMP)1-7, inhibitor of differentiation (ID) proteins ID1-3, SMAD1-9, and Jun, showed a markedly differential expression between the two chicken lines, relative to their respective controls. We then further predicted 24 known miRNAs that targeted BMP7 mRNA from 139 known miRNAs in the two chicken lines. Among these, six miRNAs were measured by qRT-PCR. In conclusion, this study is the first to analyze most of the genes, interactions, and regulators of the $TGF-{\beta}$ pathway in the innate immune responses of NE afflicted chickens.

당근을 첨가한 쉰다리의 발효 특성 (Fermentation Characteristics of Shindari Added with Carrot)

  • 김소연;박은진
    • 한국식품조리과학회지
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    • 제31권1호
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    • pp.9-17
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    • 2015
  • Shindari is a traditional fermented drink of Jeju in Korea, which is made with boiled barley and nuruk for short fermentation periods. This study determined chemical, microbiological, and sensory characteristics of the modified Shindari with 15% carrots as an additive (carrot Shindari), and this study compared it with a traditional Shindari as a control. After fermentation at $30^{\circ}C$ for a day, the pHs of the carrot Shindari and traditional Shindari largely decreased, and the total acidities increased in both of the Shindari. The significantly higher scores of Hunter's color values were observed more in carrot Shindari than in traditional Shindari. Also, carrot Shindari (0.4954 g/100 g) had a significantly higher content of vitamin C than traditional Shindari (0.0030 g/100 g). The most abundant free sugar and organic acid were glucose and lactic acid, respectively, in both of the Shindari. The total numbers of bacteria, fungi and lactic-acid bacteria in both samples increased by log 3 CFU/mL after fermentation. Based on 16S ribosomal RNA gene analysis, the dominant lactic-acid bacteria was Pediococcus acidilactici in both samples. The DPPH (1.1-diphenyl-2-picrylhydrazyl) radical scavenging activity of carrot Shindari (60.13%) was higher than that of traditional Shindari (23.70%). In sensory evaluations (taste, flavor, color, and overall acceptance), the carrot Shindari had higher scores in all these values. In this study, the modified Shindari with carrot presenting high sensory characteristic as well as chemical and microbiologic characteristics provide an opportunity to improve the application of a traditional fermented drink of Jeju, Shindari.

일측성 편마비를 동반한 A군 ${\beta}$-용혈성 사슬알균 수막뇌염 1례 (Group A ${\beta}$-hemolytic Streptococcal Meningoencephalitis with Unilateral Hemiparesis)

  • 김기원;순유진;차병호;이해용;어영;김선주;전진경
    • Pediatric Infection and Vaccine
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    • 제16권2호
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    • pp.210-214
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    • 2009
  • 저자들은 건강하던 8세 남아에서 A군 $\beta$-용혈성 사슬알균(Group A $\beta$-hemolytic streptococci, GABHS) 감염 후 발생한 일련의 면역반응으로 인한 응고장애와 좌측 편마비 및 경련이 발생한 수막뇌염을 경험하여 문헌고찰과 함께 보고하는 바이다.

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