• Title/Summary/Keyword: $E_2$

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Molecular Cloning and Expression of Human Dihydrolipoamide Dehydrogenase-Binding Protein in Excherichia coli

  • Lee, Jeong-Min;Ryou, Chong-Suk;Kwon, Moo-Sik
    • Journal of Microbiology and Biotechnology
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    • v.11 no.4
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    • pp.592-597
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    • 2001
  • The pyruvate dehydrogenase complex (PDC) catalyzes the oxidative decarboxylation of pyruvate with the formation of $CO_2$, acetyl-CoA, NADH, and H+. This complex contains multiple copies of three catalytic components including pyruvate dehydrogenase(E1), dihydrolipoamide acetyltransferase(E2), and dihydrolipoamide dehydrogenase (E3). Two regulatory components (E1-kinase and phospho-E1 phosphatase) and functionally less-understood protein (protein X, E3BP) are also involved in the formation of the complex. In this study, cloning and characterization of a gene for human E3BP have been carried out. A cDNA encoding the human E3BP was isolated by database search and cDNA library screening. The primary structure of E3BP has some similar characteristics with that of E2 in the lipoyl domain and the carboxyl-terminal domain, based on the nucleotide sequence and the deduced amino acid sequence. However, the conserved amino acid moiety including the histidine residue for acetyltransferase activity in E2 is not conserved in the case of human E3BP. The human E3BP was expressed and purified in E. coli. The molecular weight of the protein, excluding the mitochondrial target sequence, was about 50 kDa as determined by SDS-PAGE. Cloning of human E3BP and expression of the recombinant E3BP will facilitate the understanding of the role(s) of E3BP in mammalian PDC.

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Relationship among e-Service Quality, Relationship Quality, and e-Loyalty of Small Medical Clinic (소형병원의 e-서비스품질, 관계의 질, e-충성도의 영향관계)

  • Kim, JI-Young
    • Journal of the Korean Applied Science and Technology
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    • v.38 no.3
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    • pp.689-699
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    • 2021
  • The spread of the COVID-19 pandemic has been increasing non-face-to-face activities; as a result, this has resulted in the number of individuals obtaining medical information from the websites and mobile contents of medical institutions increasing. The study conducted the structural equation modeling to test hypotheses; as a result, all sub-factors of e-service quality of small medical clinic websites and mobile contents, usability, security, responsiveness, design, and information, had a significant positive effect on relationship quality, and relationship quality had a significant positive effect on e-Loyalty. Moreover, the structural equation model showed a good model fit, χ2/df of 2.021, NFI of .954, TLI of .969, CFI of .976, RMSEA of .046. Future research is suggested to study relationship quality by developing a system able to quickly and accurately respond to websites and mobile contents users; furthermore, improving e-service quality and relationship quality is likely to strengthen e-loyalty.

Strategy for V2E Performance Assurance Technology Development Using the Kano Model (Kano 모델을 활용한 V2E 성능확보기술 개발 전략)

  • Jang, Jeong Ah;Son, Sungho;Lee, Jung Ki
    • Journal of Auto-vehicle Safety Association
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    • v.14 no.2
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    • pp.75-82
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    • 2022
  • Automated vehicles (AVs) are coming to our roadways. In practice, there are still several challenges that can impede the AV sensors are polluted on various road conditions. In this paper, we propose a strategy for V2E performance assurance technology using Kano model. We are developing the vehicle sensor cleaning system about the three types of commonly used sensors: camera, radar, and LiDAR. Surveys were carried out in 30 AV's experts on quality characteristics about V2E performance assurance technology. As a result, the Kano model developed to verify a major requirement of autonomous vehicle's sensor cleaning system. It is expected that the Kano model will be actively used to verify the importance of V2E development strategy.

Effect of E-box and E2F Binding Site on Transcriptional Activity in MCM Promoter (MCM promoter에서 E-box와 E2F 결합부위가 전사활성에 미치는 영향)

  • 권현주
    • Journal of Life Science
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    • v.14 no.5
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    • pp.732-740
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    • 2004
  • MCM proteins are essential for eukaryotic DNA replication, playing roles in the initiation and elongation of DNA replication. MCM proteins expression is much higher in malignant tissues than normal tissues. Several reports have indicated the usefulness of MCM proteins as markers of cancer cells in histopathological diagnosis. However, the cause of enhanced expression of MCM proteins in cancer cells remain to be clarified. The purpose of this study is to examine the relative transcriptional activities of human mcm gene promoters in cancer and normal cells. The minimal promoter region required for transcription of a luciferase reporter gene was contained an E-box and one E2F site. In addition, luciferase activities from mcm7 and mcm2 promoter/luciferase gene reporter constructs were significantly increased in cancer cells at 8 times compared with normal cells. E-box and E2F binding site in the promoter of mcm genes are responsible for different mechanism of transcription regulation on the cellular environment.

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.

Selective Fluidization of Synaptosomal Plasma Membrane Vesicles by 17β-Estradiol

  • Lee, Sae A;Park, Yong Jin;Jang, Il Ho;Kang, Jung Sook
    • Biomedical Science Letters
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    • v.23 no.1
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    • pp.17-24
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    • 2017
  • Estrogens are effective neuroprotectants in vivo and in vitro. To obtain a better insight into the molecular mechanisms of action of neuroprotection by $17{\beta}-estradiol$ (E2), we examined the differential effects of E2 on the fluidity of synaptosomal plasma membrane vesicles (SPMV) isolated from rat cerebral cortex. Intramolecular excimerization of 1,3-di(1-pyrenyl)-propane (Py-3-Py) was used to investigate the effects of E2 on the bulk and annular lateral diffusion of the SPMV. In addition, we examined the effects of E2 on the rotational diffusion of individual leaflet of SPMV exploiting selective quenching of outer monolayer 1,6-diphenyl-1,3,5-hexatriene (DPH) fluorescence by trinitrophenyl groups. The $F{\ddot{o}}rster$ distance $R_0$ value for the tryptophan-Py-3-Py donor-acceptor pair was $26.9{\AA}$. E2 increased the lateral mobility of both bulk and annular lipids in SPMV in a dose-dependent manner, but a larger effect on bulk lipids was observed. Although E2 decreased the anisotropy of DPH in SPMV, E2 had a greater fluidizing effect on the outer leaflet compared to the inner leaflet. These results suggest that E2 selectively fluidizes the more fluid regions within SPMV. It is highly probable that E2 mostly fluidizes the bulk lipids, away from either annular lipids or lipid rafts, in the outer leaflet of SPMV. This selective fluidization may be one of the nongenomic mechanisms of neuroprotection by E2.

Clinical Implication of Prostaglandin E2 Content in Lumbar Disc Disease (추간판내 Prostaglandin E2 양의 임상적 의의)

  • Kim, Hyung Seok;Cho, Ki Hong;Kim, Ki Young;Ahn, Young Hwan;Ahn, Young Min;Yoon, Soo Han;Cho, Kyung Gi
    • Journal of Korean Neurosurgical Society
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    • v.29 no.10
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    • pp.1340-1344
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    • 2000
  • Objective : A prospective biochemical assay of prostaglandin $E_2$ content in symptomatic lumbar disc materials was done in order to clarify the pathogenesis of lumbar radiculopathy. Patients and Methods : Forty-eight disc specimens were purified by a standard solid-phase extraction procedure and analyzed by an enzymelinked immunosorbent assay for prostaglandin $E_2$. Clinical and anatomic correlations were evaluated with analysis of variance and t-test. Results : Acute herniated lumbar disc diseases tended to be associated with a higher prostaglandin $E_2$ content than degenerative lumbar disc disease. Sequestered discs tended to be associated with a higher prostaglandin $E_2$ content than extruded discs, which also showed higher prostaglandin $E_2$ content than protruded ones. A sciatica and positive straight leg raising test appeared to be associated with a higher prostaglandin $E_2$ content than a negative test. Conclusion : This result suggests that the level of prostaglandin $E_2$ would be correlated with clinical symptom and sign in the inflammatory process of lumbar disc herniation.

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Situation of HPV16 E2 Gene Status During Radiotherapy Treatment of Cervical Carcinoma

  • Kahla, Saloua;Kochbati, Lotfi;Maalej, Mongi;Oueslati, Ridha
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.6
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    • pp.2869-2873
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    • 2014
  • Background: Human papillomavirus (HPV) integration within the E2 gene has been proposed as a critical event in cervical carcinogenesis. This study concerned whether HPV16 status and E2 gene intactness are predictive of radiation response in patients with cervical cancer. Materials and Methods: Biopsies of 44 patients with cervical cancer were collected before or after radiotherapy. The presence of HPV16 was assessed by polymerase chain reaction (PCR) using specific primers for the L1 region. E2 disruption was detected by amplifying the entire E2 gene. Results: HPV16 DNA was found in 54.5% of the clinical samples. Overall, 62.5% of the HPV16 positive tumors had integrated viral genome and 37.5% had episomal genome. There was a tendency of increase of HPV16 E2 negative tumors compared with HPV16 L1 ones in advanced stages (75% versus 20% in stage III respectively). Detection of E2 gene appeared influenced by the radiotherapy treatment, as the percentage of samples containing an intact HPV16 E2 was more frequent in pretreated patients compared to radiotherapy treated patients (66.6% versus 20%). The radiation therapy caused an eight-fold [OR= 8; CI=1.22-52.25; p=0.03] increase in the risk of HPV16 genome disruption. The integration status is influenced by the irradiation modalities, interestingly E2 disruption being found widely after radiotherapy treatment (75%) with a total fractioned dose of 50Gy. Conclusions: This study reveals that the status of the viral DNA may be used as a marker to optimize the radiation treatment.

Evaluation of the virulence genes and Shiga toxin-producing abilities of Escherichia coli field isolates causing edema disease in pigs (국내 분리 돼지 부종병 대장균의 병원성 유전자 및 시가독소 생성 검증)

  • Seo, Byoung-Joo;Jeong, Chang-Gi;Kang, A-Rum;Cho, Ho-Seong;Kim, Won-Il
    • Korean Journal of Veterinary Service
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    • v.39 no.2
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    • pp.87-92
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    • 2016
  • Porcine edema disease (ED) is a communicable disease of pigs caused by infection with Shiga toxin (Stx)-producing Escherichia coli (STEC) which expresses F18 fimbriae and/or Stx type 2e (Stx2e). While STEC causes a severe illness including hemorrhagic colitis and hemolytic-uremic syndrome in humans, it induces damage to the vascular endothelium, which results in edema, hemorrhage, and microthrombosis, leading in high mortality in pigs. In the present study, we cultured Stx2e-producing E. coli field isolates from conventional pig farms that experienced sudden deaths previously with symptoms similar to porcine edema disease, which were further investigated with Shiga toxin profiles. A total of 43 strains were identified from the collected samples by F18 or Stx2e specific PCR. Based on the PCR, 42 isolates out of 43 isolates were proved to carry one of F18 or Stx2e genes and 14 isolates to carry both F18 and Stx2e genes. All of the 30 isolates that harbored Stx2e gene induced the cytopathic effect (CPE) in vero cells and especially, the isolate 150229 produced the highest level of Shiga toxin. Therefore, we identified the virulence genes (F18 and Stx2e) and demonstrated Shiga toxin-producing abilities from porcine edema disease causing E. coli filed isolates. These results suggested that one of the isolates could be a vaccine antigen candidate against STEC through further investigating to elicit an immune response.

Accuracy of the Position Fix and Local Fluctuation of Omega Signals in the South Coast of Korea (한국남해안에 있어서의 오메가신호의 지역적인 변화와 측위의 정도)

  • 안장영
    • Journal of the Korean Society of Fisheries and Ocean Technology
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    • v.17 no.2
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    • pp.59-63
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    • 1981
  • For the purpose of surveying accuracy of the position fix and local fluctuation of Omega signals in south coast of Korea, the author observe continuously the phase difference of the A-C pair and A-E pair form December 1, 1980, to February 28, 1981, at Cheju(33$^{\circ}$31'.0N, 126$^{\circ}$31.9E) and Busan(35$^{\circ}$7'.9 N, 129$^{\circ}$6'. 3 E). The analyzed results are as follows: 1. Omega signals at Cheju and Busan are positive correlation. Mean value of coefficients during all measured time is 0.70 with A-C pair and 0.75 with A-E pair. 2. Range of mean value of difference between the measured PPC at Cheju and Busan is 17.7~21.3 cels with A-C pair and -2.9~2.0 cels with A-E pair. 3. In case of using the differential Omega at Cheju and Busan, mean value of total errors of the position fix is 1.52 miles and its standard deviation is 0.21 miles.

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