• Title/Summary/Keyword: MEP

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Biosynthesis of Isoprenoids: Characterization of a Functionally Active Recombinant 2-C-methyl-D-erythritol 4-phosphate Cytidyltransferase (IspD) from Mycobacterium tuberculosis H37Rv

  • Shi, Wenjun;Feng, Jianfang;Zhang, Min;Lai, Xuhui;Xu, Shengfeng;Zhang, Xuelian;Wang, Honghai
    • BMB Reports
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    • v.40 no.6
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    • pp.911-920
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    • 2007
  • Tuberculosis, caused by Mycobacterium tuberculosis, continues to be one of the leading infectious diseases to humans. It is urgent to discover novel drug targets for the development of antitubercular agents. The 2-C-methyl-Derythritol-4-phosphate (MEP) pathway for isoprenoid biosynthesis has been considered as an attractive target for the discovery of novel antibiotics for its essentiality in bacteria and absence in mammals. MEP cytidyltransferase (IspD), the third-step enzyme of the pathway, catalyzes MEP and CTP to form 4-diphosphocytidyl-2-C-methylerythritol (CDP-ME) and PPi. In the work, ispD gene from M. tuberculosis H37Rv (MtIspD) was cloned and expressed. With N-terminal fusion of a histidine-tagged sequence, MtIspD could be purified to homogeneity by one-step nickel affinity chromatography. MtIspD exists as a homodimer with an apparent molecular mass of 52 kDa. Enzyme property analysis revealed that MtIspD has high specificity for pyrimidine bases and narrow divalent cation requirements, with maximal activity found in the presence of CTP and $Mg^{2+}$. The turnover number of MtIspD is $3.4 s^{-1}$. The Km for MEP and CTP are 43 and $92{\mu}M$, respectively. Furthermore, MtIspD shows thermal instable above $50^{\circ}C$. Circular dichroism spectra revealed that the alteration of tertiary conformation is closely related with sharp loss of enzyme activity at higher temperature. This study is expected to help better understand the features of IspD and provide useful information for the development of novel antibiotics to treat M. tuberculosis.

Molecular cloning and expression analysis of the first two key genes through 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway from Pyropia haitanensis (Bangiales, Rhodophyta)

  • Du, Yu;Guan, Jian;Xu, Ruijun;Liu, Xin;Shen, Weijie;Ma, Yafeng;He, Yuan;Shen, Songdong
    • ALGAE
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    • v.32 no.4
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    • pp.359-377
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    • 2017
  • Pyropia haitanensis (T. J. Chang et B. F. Zheng) N. Kikuchi et M. Miyata is one of the most commercially useful macroalgae cultivated in southeastern China. In red algae, the biosynthesis of terpenoids through 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway can produce a direct influence on the synthesis of many biologically important metabolites. In this study, two genes of cDNAs, 1-deoxy-D-xylulose-5-phosphate synthase (DXS) and 1-deoxy-D-xylulose-5-phosphate reductase (DXR), which encoding the first two rate-limiting enzymes among MEP pathway were cloned from P. haitanensis. The cDNAs of P. haitanensis DXS (PhDXS) and DXR (PhDXR) both contained complete open reading frames encoding polypeptides of 764 and 426 amino acids residues, separately. The expression analysis showed that PhDXS was significant differently expressed between leafy thallus and conchocelis as PhDXR been non-significant. Additionally, expression of PhDXR and its downstream gene geranylgeranyl diphosphate synthase were both inhibited by fosmidomycin significantly. Meanwhile, we constructed types of phylogenetic trees through different algae and higher plants DXS and DXR encoding amino acid sequences, as a result we found tree clustering consequences basically in line with the "Cavalier-Smith endosymbiotic theory." Whereupon, we speculated that in red algae, there existed only complete MEP pathway to meet needs of terpenoids synthesis for themselves; Terpenoids synthesis of red algae derivatives through mevalonate pathway came from two or more times endosymbiosis of heterotrophic eukaryotic parasitifer. This study demonstrated that PhDXS and PhDXR could play significant roles in terpenoids biosynthesis at molecular levels. Meanwhile, as nuclear genes among MEP pathway, PhDXS and PhDXR could provide a new way of thinking to research the problem of chromalveolata biological evolution.

Chemical Resistance of Striped Rice Borer, Chilo Suppressalis, and Green Rice Leafhopper, Nephotettix cincticeps (이화명충과 끝동매미충의 약제저항성)

  • Lee Seung Chan;Yoo Jai Ki
    • Korean journal of applied entomology
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    • v.14 no.2 s.23
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    • pp.65-70
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    • 1975
  • The study was planned to detect resistance levels of striped rice borers (Chilo suppressalis Walker) and green rice leafhoppers(Nephotettix cincticeps Uhler) from different localities to organophosphate and carbamate insecticides using topical application. Strains of overwintering striped rice borers were collected from 7 different areas in Gyeonggi Province and they were kept under conditions of about $25^{\circ}C$ for 10 days. Overwintered and reared larvae of striped rice borers were compared for response to insecticides. The strains of green rice leafhopper were collected from Iri, Milyang, Pyongtaek, Echeon and Suweon. Insects were multiplied in the lab. All insecticides tested were organphosphates(MPP, MEP and Diazinon) and carbamate (NAC), which have been used in control of rice insect pests for over past 10 years. The results obtained were as follows; a. With MPP compound, resistant levels of the Joam, Suweon, and Echeon strains of C. suppressalis were from 2 to 4 times as compared with the Yangpyong strain. b. With MEP insecticide, resistant levels of the Kimpo, Pyongtaek, Yongin, Echeon, Suweon, and Joam strains of C. suppressalis were from 2 to 6 times as compared with the Yangpyong strain. c. With Diazinon, resistant levels of the Suweon strain of C. suppressalis were 3 times greater than the Yangpyong strain; but the resistant levels of Kimpo, Pyongtaek, Yongin, and Echeon strains approximated the later. 4. The overwintered larvae of C. suppressalis were more tolerant to the insecticides than the larvae reared in the lab. e. With MEP, resistant levels of the Iri, Milyang, and Pyongtaek strains of N. cincticeps to the Suweon strain showed 13, 8, 7 and4 times, respectively. f. With MPP, resistant levels of the Iri and Pyongtaek strains of N. cincticeps were 7 and 4 times as compared with the Suweon strain, respectively. g. With NAC, resistant levels of the several strains of if. cincticeps showed no difference, and seemed not to have developed resistance yet.

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Consideration of Ruptured Cerebral Aneurysms Clipping Surgery

  • Park, Sang-Ku;Kim, Dong-Jun;Han, Hung-Tae
    • Korean Journal of Clinical Laboratory Science
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    • v.47 no.1
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    • pp.24-27
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    • 2015
  • To measure motor evoked potentials (MEP) during emergency surgery is often difficult in patients with subarachnoid hemorrhage (SAH) from a ruptured cerebral aneurysm, The cause of these difficulties may be considered as damage to the motor pathway by hemorrhage. To identify the cause of difficulties in measuring MEP, we defined the association between motor evoked potentials during surgery and the severity of the hemorrhage in patients with subarachnoid hemorrhage.

Effects of Yukmijihwang-tang on Maximal Exercise Performance (육미지황탕 투여가 운동 지구력 향상에 미치는 영향)

  • Bae, Gi Sang;Park, Jin Han
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.29 no.6
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    • pp.498-502
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    • 2015
  • Yukmijihwang-tang (YM) is a well-known prescription to treat kidney inflammation on Korea. However, the effects of YM on maximal exercise performance (MEP) is not-well investigated. In this study, we aimed to evaluate the effects of YM on MEP using forced swimming test (FST) model. Mice were largely divided into three group; a non-swimming group, saline treated-swimming load test group, and YM treated-swimming load group. Swimming load test groups were re-divided into swimming control group (Control), YM fed group (YM). YM was administrated orally for 2 weeks before FST. After FST, immobility time, oxygen consumption was measured by physiological test, and serum was collected for biochemical analysis. FST induced MEP identified by increased immobility, oxygen consumption, and biochemical factors. Immobility time was significantly reduced by YM treatment compared with control group. Oxygen consumption was also significantly reduced by YM treatment. The increase on lactic acid and lactate dehydrogenase after FST was inhibited by YM treatment. In addition, consumption of energy source (free fatty acid, and triglyceride) and recovery of energy was enhanced by YM treatment after FST. In conclusion, YM inhibited the elevation of immobility time, oxygen consumption, and biochemical factors after FST. YM could be used as a beneficial agents to enhance maximal endurance exercise performance and an effective ergogenic aid in exercise training.

Investigation of Pesticide Residues in Fruits and Vegetables (과채류(果菜類) 중(中) 농약잔류(農藥殘留) 수준조사연구(水準調査硏究))

  • Rhu, H.I.;Suh, Y.S.;Kim, I.K.;Kim, H.Y.;Jun, S.H.
    • Korean Journal of Environmental Agriculture
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    • v.6 no.1
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    • pp.61-66
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    • 1987
  • 1. In the vegetable crops, only MEP and Malathion showed a litte higher positive rate than other pesticides in general. On the other hand, parathion and captan were not detectable at all. 2. In the fruits, pesticides, MEP, PAP and Parathion had a little high positive rate, and Diazinon, and captan were not detectable at all. 3. However, the detected values of the pesticides in the vegetables and fruits were far below from korean tolerances and internationally recommended limits and tolerances, showing only $\frac{1}{3}{\sim}\frac{1}{66}$ and $\frac{1}{8}{\sim}\frac{1}{500}$ of Korean tolerance in vegetables and fruits, respectively.

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Technical Considerations of Effective Direct Cortical and Subcortical Stimulation (효과적인 대뇌 직접피질자극 검사 및 피질하자극 검사의 술기에 관한 기술적 고찰)

  • Lim, Sung Hyuk;Jang, Min Hwan
    • Korean Journal of Clinical Laboratory Science
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    • v.54 no.2
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    • pp.157-162
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    • 2022
  • The purpose of the direct cortical and subcortical stimulation technique is to prevent false positives caused by transcranial electrical motor evoked potentials (TceMEP) in surgery on patients with brain tumors that have occurred around the motor cortex and to preserve the correct mapping of motor areas during surgery and the corticospinal tract. In addition, it reduces the trial and error that occurs during the intraoperative neurophysiological monitoring (INM) process and minimizes the test time, so that accurate information is communicated to the surgeon with quick feedback on the test results. The most important factors of this technique are, first, examination at a stimulus threshold of a certain intensity, and second, maintaining anesthesia depth at an appropriate level to prevent false positives from occurring during surgery. The third is the installation of a multi-level channel recording electrode on the opposite side of the area of operation to measure the TceMEP waveform and the response to direct cortical and subcortical stimulation in as many muscles as possible. If these conditions are maintained, it is possible to predict causes that may occur in other factors, not false positives, from the INM test.

Vocal Characteristics and Differences in Gender and Voice Classification among Classical Singers (성악가의 성별 및 성종에 따른 발성적 특징과 차이)

  • Nam, Do-Hyun;Kim, Wha-Soak
    • Phonetics and Speech Sciences
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    • v.1 no.2
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    • pp.163-171
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    • 2009
  • This study attempted to investigate vocal characteristics and differences in gender and voice classification among classical singers. Twenty-three female singers (M = 23.1 yrs, SD = 3.6 yrs, average 6.3 yrs singing experience, all classified as sopranos) and twenty male singers (M = 25.2 yrs, SD= 3.6 yrs, average 6. 3 yrs singing experience, 8 tenors, 12 baritones) were recruited to participate in the present study. Speaking fundamental frequency (FO), closed quotient (CQ), MPT (Maximum Phonation Time), breathing types, maximum inspiratory pressure (MIP), maximum expiratory pressure (MEP), and singers' formants were measured. In addition, vibratory patterns were observed using stroboscopy. Sfo, singing CQ, breathing types, formant frequency in singers' formants, MIP, MEP, and MPT were significantly different from gender to gender. Generally, singers' formants were observed in male singers and also the pattern of singers' formants was different between tenors and baritones. Lower singing CQ values were observed than speaking CQ values in the female singers (P<.001). Furthermore, MEP, MIP, and singing CQ were significantly lower for female singers than for males singers (P<.001). MPT and speaking FO, however, were not significantly different between tenors and baritones.

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Highly Integrated Low-Power Motion Estimation Processor for Mobile Video Coding Applications (이동통신 향 동영상압축을 위한 고집적 저전력 움직임 추정기)

  • Park Hyun Sang
    • Journal of Broadcast Engineering
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    • v.10 no.1 s.26
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    • pp.77-82
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    • 2005
  • We propose a highly Integrated motion estimation processor (MEP) for efficient video compression in an SoC platform. When compressing video by the standards like MPEG-4 and H.263, the macroblock related functions motion compensation. mode decision, motion vector prediction, and motion vector difference calculation require the frequent intervention of MCU. Thus the proposed MEP incorporates those functions with the motion estimation capability to reduce the number of interrupts to MCU, which can lead to a highly efficient SoC system. For low-power consumption, the proposed MEP can prevent the temporally static area from motion estimation or can skip the half-pel motion estimation for those macroblocks whose modes are decided as INTRA.

Comparative Modeling Studies of 1-deoxy-D-xylulose 5-phosphate Synthase (MEP pathway) from Mycobacterium Tuberculosis

  • Kothandan, Gugan
    • Journal of Integrative Natural Science
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    • v.4 no.3
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    • pp.202-209
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    • 2011
  • Tuberculosis is a major health problem in humans because of its multidrug resistance and discovering new treatments for this disease is urgently required. The synthesis of isoprenoids in Mycobacterium tuberculosis has been reported as an interesting pathway to target. In this context, 2C-methyl-D-erythritol 4-phosphate (MEP) pathway of M. tuberculosis has drawn attention. The MEP pathway begins with the condensation of glyceraldehyde 3-phosphate and pyruvate forming 1-deoxy-D-xylulose 5-phosphate (DXP) which is catalyzed by 1-deoxy-D-xylulose 5-phosphate synthase (DXS). As there is no X-ray structure was reported for this target, comparative modeling was used to generate the three dimensional structure. The structure was further validated by PROCHECK, VERIFY-3D, PROSA, ERRAT and WHATIF. Molecular docking studies was performed with the substrate (Thiamine pyrophosphate) and the reported inhibitor 2-methyl-3-(4-fluorophenyl)-5-(4-methoxy-phenyl)-4H-pyrazolol[1,5-a]pyrimidin-7-one) against the developed model to identify the crucial residues in the active site. This study may further be useful to provide structure based drug design.