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Evaluation of Selective Media Containing Iron Source and Alpha-Glucosidase Substrates for Enterobacter sakazakii (Cronobacter spp.) Detection

  • Chon, Jung-Whan;Seo, Kun-Ho;Yim, Jin-Hyeok;Bae, Dongryeoul;Kim, Binn;Kim, Tae-Jin;Jeong, Dongkwan;Song, Kwang-Young
    • Journal of Dairy Science and Biotechnology
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    • v.39 no.1
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    • pp.9-19
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    • 2021
  • Enterobacter sakazakii (Cronobacter spp.) causes meningitis, necrotizing enterocolitis, sepsis, and bacteremia in neonates and children and has a high mortality rate. For rapid E. sakazakii detection, various differential and selective media containing α-glucosidase substrates, such as 5-bromo-4-chloro-3-indolyl-α-D-glucopyranoside (BCIG) or 4-methylumbelliferyl-α-D-glucoside (α-MUG), have been developed as only E. sakazakii exhibits α-glucosidase activity in the genus Enterobacter. However, Escherichia vulneris (family: Enterobacteriaceae) can also utilize α-glucosidase substrates, thereby resulting in false positives. Various iron sources are known to promote the growth of gram-negative bacteria. This study aimed to develop a selective medium containing α-glucosidase substrates for E. sakazakii detection that would eliminate false positives, such as those of E. vulneris, and to determine the role of iron source in the medium. Three previously developed (TPD) media, i.e., Oxoid, OK, and VRBG, and the medium developed in this study, i.e., NGTE, were evaluated using 58 E. sakazakii and 5 non-E. sakazakii strains. Fifty-four E. sakazakii strains appeared as fluorescent or chromogenic colonies on all four media that were assessed. Two strains showed colonies on NGTE medium and not on TPD media. In contrast, the remaining two strains showed colonies on TPD media and not on NGTE medium. None of the non-E. sakazakii strains showed fluorescent or chromogenic colonies on any of the evaluated media except E. vulneris, which showed colonies on TPD media and not on NGTE medium. This study demonstrated that the newly developed NGTE medium was not only equally efficient in promoting the growth of bacterial colonies when compared with the currently available media but also eliminated false positives, such as E. vulneris.

Co-Infection with Cytomegalovirus and Helicobacter pylori in a Child with $M\acute{e}n\acute{e}$trier's Disease

  • Yoo, Yangho;Lee, Yoon;Lee, Yoo Min;Choe, Yon Ho
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.16 no.2
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    • pp.123-126
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    • 2013
  • $M\acute{e}n\acute{e}$trier's disease is a rare protein-losing gastropathy characterized by hypertrophic gastric fold, foveolar hyperplasia, and hypoproteinemia with resulting peripheral edema. It is clinically evident as nonspecific gastrointestinal symptoms, including abdominal discomfort, nausea and vomiting, abdominal pain, weight loss, diarrhea, and edema. Pediatric $M\acute{e}n\acute{e}$trier's disease usually has an insidious onset and progressive, chronic clinical course and it spontaneously resolves in weeks or months. The pathogenesis of $M\acute{e}n\acute{e}$trier's disease is not clearly understood. $M\acute{e}n\acute{e}$trier's disease is thought to be associated with some gastric infections. But the cause of $M\acute{e}n\acute{e}$trier's disease is unknown, an association with cytomegalovirus (CMV) and Helicobacter pylori has been suggested. In Korea, We present the first a case of pediatric $M\acute{e}n\acute{e}$trier's disease with positive evidence of CMV and H. pylori.

Studies on Changes of the Droplets by Bubbles in Piezoelectric Inkjet Head (잉크젯 헤드내 발생한 기포에 따른 토출 변화 연구)

  • Yoo, Young-Seuck;Kim, Young-Jae;Sim, Won-Chul;Park, Chang-Sung;Park, Jung-Hoon;Kang, Pil-Joong;Joung, Jae-Woo;Oh, Yong-Soo
    • Proceedings of the KIEE Conference
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    • 2007.07a
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    • pp.1544-1545
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    • 2007
  • 본 논문은 피에조방식으로 구동하는 MEMS 구조의 산업용 잉크젯 헤드를 제작하여 잉크를 충진하여 토출하는 과정에서 토출이 되지 않는 원인 중 하나인 기포에 대해서 연구하였다. 기포를 직접 관찰하기 위한 방법으로 투명한 유리로 Membrane을 제작하여 기포가 발생하여 거동하는 모습을 관찰하였으며 Actuator가 구동하는 헤드내 기포를 구동 중에 관찰하기 위한 방법으로 LDV(Laser Doffler Vibrometer)를 이용하였다. 그 결과, 구동하면서 발생하는 변위의 미세한 차이를 관찰할 수 있었으며 주파수 data의 차이를 관찰함으로써 기포의 크기에 따른 토출의 양태를 구별할 수 있었다.

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HPV 16 E6/E7 Transgenic Mice have Hyperkeratosis and Elevated ROS Related Enzyme Activities

  • Kim, Myoung-Ok;Lee, Eun-Ju;Kim, Sung-Hyun;Park, Jun-Hong;Kyoungin-Cho;Jung, Boo-Kyung;Kim, Hee-Chul;Sol ha Hwang;Kim, Sun-Jung
    • Proceedings of the KSAR Conference
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    • 2003.06a
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    • pp.45-45
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    • 2003
  • Human papillomavirus type 16(HPV16) has been known to the major factor for the development of uterine cervical carcinomas. We have extended these studies to investigate the in vivo activities of HPV-16 E6/E7 when expressed in squamous epithelia of transgenic mice. Grossly, hK14HPV16E6/E7 transgenic mice had multiple phenotypes, including wrinkled skin that was apparent prior to the appearance of hair on neonates, thickened ears, and loss of hair in adults. In the transgenic mice, the wrinkled skin phenotype on the body and legs died at the age of 3∼4 weeks. Histological analysis of demonstrated that E6/E7 causes epidermal hyperplasia in multiple transgenic lineages with high penetrance. This epithelial hyperplasia was characterized by an expansion of the proliferating compartment and an expansion of the keratinocyte and was associated with hyperkeratosis. These transgenic mice expressed E6/E7 transgene mainly in skin, heart, pancreas and kidney. Hyperplasia was found at the skin. The enzyme activities of GR, GPx and CuZnSOD were measured from the transgene cause keratinocyte at the skin. The specific enzyme activities were significantly higher in transgenic mice skin compared to the normal mice skin. Thus these transgenic mice may be useful for the develpment of antioxidant enzymes or other therapies for HPV-associated hyperkeratosis.

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Cytotoxic T Lymphocytes Elicited by Dendritic Cell-Targeted Delivery of Human Papillomavirus Type-16 E6/E7 Fusion Gene Exert Lethal Effects on CaSki Cells

  • Wu, Xiang-Mei;Liu, Xing;Jiao, Qing-Fang;Fu, Shao-Yue;Bu, You-Quan;Song, Fang-Zhou;Yi, Fa-Ping
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.6
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    • pp.2447-2451
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    • 2014
  • Human papillomavirus (HPV) is the primary etiologic agent of cervical cancer. Consideration of safety and non human leukocyte antigen restriction, protein vaccine has become the most likely form of HPV therapeutic vaccine, although none have so far been reported as effective. Since tumor cells consistently express the two proteins E6 and E7, most therapeutic vaccines target one or both of them. In this study, we fabricated DC vaccines by transducing replication-defective recombinant adenoviruses expressing E6/E7 fusion gene of HPV-16, to investigate the lethal effects of specific cytotoxic T lymphocytes (CTL) against CaSki cells in vitro. Mouse immature dendritic cells (DC) were generated from bone marrow, and transfected with pAd-E6/E7 to prepare a DC vaccine and to induce specific CTL. The surface expression of CD40, CD68, MHC II and CD11c was assessed by flow cytometry (FCM), and the lethal effects of CTL against CaSki cells were determined by DAPI, FCM and CCK-8 methods. Immature mouse DC was successfully transfected by pAd-E6/E7 in vitro, and the transfecting efficiency was 40%-50%. A DC vaccine was successfully prepared and was used to induce specific CTL. Experimental results showed that the percentage of apoptosis and killing rate of CaSki cells were significantly increased by coculturing with the specific CTL (p <0.05). These results illustrated that a DC vaccine modified by HPV-16 E6/E7 gene can induce apoptosis of CaSki cells by inducing CTL, which may be used as a new strategy for biological treatment of cervical cancer.

13-Hydroxy-9Z,11E,15E-octadecatrienoic Acid from the Leaves of Cucurbita moschata

  • Bang, Myun-Ho;Han, Jae-Taek;Kim, Hae-Yeong;Park, Young-Doo;Park, Chang-Ho;Lee, Kang-Ro;Baek, Nam-In
    • Archives of Pharmacal Research
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    • v.25 no.4
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    • pp.438-440
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    • 2002
  • A new unsaturated hydroxy fatty acid was isolated from the leaves of Cucurbita moschata through repeated silica gel column chromatography and chemical methods. The structure of the new fatty acid was determined as 13-hydroxy-9, 11, 15-octadecatrienoic acid on the basis of several spectral data including 2D-NMR. The stererostructures of double bonds were determined to be 9Z, 11 E and 15E by coupling patterns of related proton signals in the $^1H-NMR$ and NOESY experiments.

Analysis of E,E-farnesol and squalene in makgeolli using stir bar sorptive extraction coupled with gas chromatography-mass spectrometry (SBSE-GCMS를 이용한 막걸리 중의 E,E-farnesol과 squalene분석법)

  • Ha, Jaeho;Shim, You-Shin;Cho, Yongsun;Seo, Dongwon;Jang, Hyewon;Jang, Hyejin
    • Analytical Science and Technology
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    • v.27 no.1
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    • pp.60-65
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    • 2014
  • The aim of this study was to establish an analytical method for the determination of E,E-farnesol and squalene in makgeolli, which is a traditional type of Korean fermented rice wine. E,E-farnesol and squalene in makgeolli were extracted using stir bar sorptive extraction (SBSE) coupled with gas chromatography-mass spectrometry. SBSE was found to be an effective method for analyzing the E,E-farnesol and squalene levels in makgeolli. The linear dynamic range of the SBSE method for detecting E,E-farnesol and squalene ranged from 0.5 to 200 ng/mL with $R^2=0.9974$ for E,E-farnesol and 100 to 50000 ng/mL with $R^2=0.9982$ for squalene. The limit of detection and the limit of quantification using the SBSE method were 0.1 and 0.5 ng/mL for E,E-farnesol and 15.0 and 40.0 ng/mL for squalene, respectively. The average recoveries obtained were, quantitatively, 101-107% for E,E-farnesol and 98-103% for squalene, respectively, supporting the accuracy of the SBSE-GCMS method.

The relationship between the variants in the 5'-untranslated regions of equine chorionic gonadotropin genes and serum equine chorionic gonadotropin levels

  • Liu, ShuQin;Lian, Song;Yang, YunZhou;Fu, ChunZheng;Ma, HongYing;Xiong, ZhiYao;Ling, Yao;Zhao, ChunJiang
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.12
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    • pp.1679-1683
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    • 2017
  • Objective: An experiment was conducted to study the association between the single nucleotide polymorphisms (SNPs) in 5'-untranslated regions (5'-UTR) of equine chorionic gonadotropin (eCG) genes and the serum eCG levels. Methods: SNPs in 5'-UTR of eCG genes were screened across 10 horse breeds, including 7 Chinese indigenous breeds and 3 imported breeds using iPLEX chemistry, and the association between the serum eCG levels of 174 pregnant Da'an mares and their serum eCG levels (determined with ELISA) was analyzed. Results: Four SNPs were identified in the 5'-UTR of the $eCG{\alpha}$ gene, and one of them was unique in the indigenous breeds. There were 2 SNPs detected at the 5' end of the $eCG{\beta}$ subunit gene, and one of them was only found in the Chinese breeds. The SNP g.39948246T>C at the 5'-UTR of $eCG{\alpha}$ was associated significantly with eCG levels of 75-day pregnant mare serum (p<0.05) in Da'an mares. Prediction analysis on binding sites of transcription factors showed that the g.39948246T>C mutation causes appearance of the specific binding site of hepatocyte nuclear factor 3 forkhead homolog 2 (HFH-2), which is a transcriptional repressor belonging to the forkhead protein family of transcription factors. Conclusion: The SNP g.39948246T>C at the 5'-UTR of $eCG{\alpha}$ is associated with eCG levels of 75-day pregnant mare serum (p<0.05).

Fiber Drawing Induced Defects in Silica Optical Fiber (광섬유 인선 공정에 의해 생성된 실리카 광섬유내 점결함)

  • 안병길;이종원;김효태
    • Journal of the Korean Ceramic Society
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    • v.40 no.11
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    • pp.1102-1105
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    • 2003
  • The fiber drawing process induced defects in silica fiber have been investigated. This study has focused on the Oxygen Deficient Centers (ODCs) and E' centers induced by the fiberization process in low-OH silica fibers. To investigate those defects induced by the fiberization process, the optical absorption spectrum and Electron Spin Resonance (ESR) have both been employed. The concentration of Oxygen Deficient Centers (ODCs) and E' centers are increased by the fiber drawing process. The population of defects in the neck-down region has also been investigated. The most significant generation of defects during fiber drawing process has been shown to occur in this region of silica preform. The population of defects is higher on the edge region than in the center of neck-down region.

Vibrio vulnificus Metalloprotease VvpE has no Direct Effect on Iron-uptake from Human Hemoglobin

  • Sun, Hui-Yu;Han, Song-Iy;Choi, Mi-Hwa;Kim, Seong-Jung;Kim, Choon-Mee;Shin, Sung-Heui
    • Journal of Microbiology
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    • v.44 no.5
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    • pp.537-547
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    • 2006
  • This study was designed to determine whether or not Vibrio vulnificus metalloprotease VvpE can promote iron uptake via the proteolytic cleavage of human hemoglobin. We found that V. vulnificus utilized hemoglobin as an iron source more efficiently via the vulnibactin-mediated iron-uptake system than via the HupA-mediated iron-uptake system and, of the proteases produced by V. vulnificus, VvpE was found to be the only protease capable of destroying hemoglobin. However, VvpE expression, on both the transcriptional and protein levels, was suppressed in iron-limited media. However, vvpE transcription, but not extracellular VvpE production, was reactivated by the addition of hemoglobin or inorganic iron into iron-limited media. Moreover, vvpE transcription began only in the late growth phase when V. vulnificus had already consumed most of the iron for growth. In addition, neither vvpE mutation nor in trans vvpE complementation affected the ability of V. vulnificus to acquire iron or to grow in iron-limited media or in cirrhotic ascites containing hemoglobin. Hemoglobin added into iron-limited media was not destroyed, but gradually formed an insoluble aggregate during culture; this aggregation of hemoglobin occurred regardless of vvpE mutation or complementation. These results indicate that VvpE is not required for efficient iron uptake from hemoglobin. On the contrary, hemoglobin or iron is required for efficient vvpE transcription. In addition, a discrepancy exists between vvpE transcription and extracellular VvpE production in iron-limited media containing inorganic iron or hemoglobin, which suggests that additional unknown posttranscriptional events may be involved in the extracellular production of VvpE.