• Title/Summary/Keyword: $C_4-pathway$

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Kinetic Property and Phylogenie Relationship of 2-Hydroxy-muconic Semialdehyde Dehydrogenase Encoded in tomC Gene of Burkholderia cepacia G4

  • Reddy, Alavala-Matta;Min, Kyung-Rak;Lee, Kyoung;Lim, Jai-Yun;Kim, Chi-Kyung;Kim, Young-Soo
    • Archives of Pharmacal Research
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    • v.27 no.5
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    • pp.570-575
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    • 2004
  • 2-Hydroxymuconic semialdehyde (2-HMS) dehydrogenase catalyzes the conversion of 2-HMS to 4-oxalocrotonate, which is a step in the meta cleavage pathway of aromatic hydrocarbons in bacteria. A tomC gene that encodes 2-HMS dehydrogenase of Burkholderia cepacia G4, a soil bacterium that can grow on toluene, cresol, phenol, or benzene, was overexpressed into E. coli HB 101, and its gene product was characterized in this study. 2-HMS dehydrogenase from B. cepacia G4 has a high catalytic efficiency in terms of V$_{max}$K$_{max}$ towards 2-hydroxy-5-methyl-muconic semialdehyde followed by 2-HMS but has a very low efficiency for 5-chloro-2-hydroxymuconic semialdehyde. However, the enzyme did not utilize 2-hydroxy-6-oxo-hepta 2,4-dienoic acid and 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoic acid as substrates. The molecular weight of 2-HMS dehydrogenase from B. cepacia G4 was predicted to be 52 kDa containing 485 amino acid residues from the nucleotide sequence of the tomC gene, and it exhibited the highest identity of 78% with the amino acid sequence of 2-HMS dehydrogenase that is encoded in the aphC gene of Comamonas testosteroni TA441. 2-HMS dehydrogenase from B. cepacia G4 showed a significant phylogenetic relationship not only with other 2-HMS dehydrogenases, but also with different dehydrogenases from evolutionarily distant organisms.sms.

Characteristic Intracelluar Response to Lidocaine And MK-801 of Hippocampal Neurons: An In Vivo Intracellular Neuron Recording Study

  • Choi, Byung-Ju;Cho, Jin-Hwa
    • The Korean Journal of Physiology and Pharmacology
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    • v.2 no.3
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    • pp.297-305
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    • 1998
  • This study used in vivo intracellular recording in rat hippocampus to evaluate the effect of lidocaine and MK-801 on the membrane properties and the synaptic responses of individual neurons to electrical stimulation of the commissural pathway. Cells in control group typically fired in a tonic discharge mode with an average firing frequency of $2.4{\pm}0.9$ Hz. Neuron in MK-801 treated group (0.2 mg/kg, i.p.) had an average input resistance of $3.28{\pm}5.7\;M{\Omega}$ and a membrane time constant of $7.4{\pm}1.8$ ms. These neurons exhibited $2.4{\pm}0.2$ ms spike durations, which were similar to the average spike duration recorded in the neurons of the control group. Slightly less than half of these neurons were firing spontaneously with an average discharge rate of $2.4{\pm}1.1$ Hz. The average peak amplitude of the AHP following the spikes in these groups was $7.4{\pm}0.6$ mV with respect to the resting membrane potential. Cells in MK-801 and lidocaine treated group (5 mg/kg, i.c.v.) had an average input resistance of $3.45{\pm}6.0\;M{\Omega}$ and an average time constant of $8.0{\pm}1.4$ ms. The cells were firing spontaneously at an average discharge rate of $0.6{\pm}0.4$ Hz. Upon depolarization of the membrane by 0.8 nA for 400 ms, all of the tested cells exhibited accommodation of spike discharge. The most common synaptic response contained an EPSP followed by early-IPSP and late-IPSP. Analysis of the voltage dependence revealed that the early-IPSP and late-IPSP were putative $Cl^--and\;K^+-dependent$, respectively. Systemic injection of the NMDA receptor blocker, MK-801, did not block synaptic responses to the stimulation of the commissural pathway. No significant modifications of EPSP, early-IPSP, or late-IPSP components were detected in the MK-801 and/or lidocaine treated group. These results suggest that MK-801 and lidocaine manifest their CNS effects through firing pattern of hippocampal pyramidal cells and neural network pattern by changing the synaptic efficacy and cellular membrane properties.

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Nitroxergic Nerve Relaxes Rat Gastric Smooth Muscle by NO-cGMP Pathway

  • Yoon, Yoong-Sam;Choi, Hyoung-Chul;Jung, Young-Sook;Kim, Jong-Ho;Lee, Kwang-Youn;Sohn, Uy-Dong;Ha, Jeoung-Hee;Kim, Won-Joon
    • The Korean Journal of Physiology and Pharmacology
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    • v.4 no.5
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    • pp.369-378
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    • 2000
  • This study was undertaken to investigate an involvement of nitroxergic innervation in gastric smooth muscle of rat. Isometric tension study, the measurement of single cell length, NADPH diaphorase stain of smooth muscle layers and neuronal nitric oxide synthase (nNOS) western blotting were performed. Sodium nitroprusside (SNP), a nitric oxide donor, relaxed the muscle strips precontracted by acetylcholine (ACh) in a concentration-dependent manner. Pretreatment of L-arginine decreased the contraction induced by electric field stimulation (EFS). Pretreatment of $N^G-nitro-L-arginine$ methyl ester (L-NAME), a NOS inhibitor, increased the EFS-induced contractions. LY 83583, a guanylate cyclase (GC) inhibitor, reversed the inhibitory actions of L-arginine on the muscle contractions. The effects of L-Arginine, L-NAME and LY 83583 on ACh-induced contractions were not significant. L-arginine reduced the EFS-induced contraction in circular muscle, whereas L-NAME enhanced the EFS-induced contraction in longitudinal strips. By EFS, the phasic contractions appeared approximately $20{\sim}25$ seconds later. L-NAME significantly shortened the delay time to about $2{\sim}3$ seconds. In single cell study, ACh contracted gastric smooth muscle cells, SNP relaxed the cells, and the latter also inhibited the ACh-induced contraction. LY 83583 enhanced the ACh-induced contraction and antagonized SNP-induced relaxation. NADPH diaphorase activity was assessed by a histochemistry, nitroblue tetrazolium (NTB) staining. Positive staining was observed in both circular and longitudinal muscle layers. L-arginine increased the staining, while L-NAME decreased the staining. Western blotting for nNOS proved the presence of nNOS in rat gastric smooth muscle. EFS and additional $Ca^{2+}$ increased nNOS protein expression. These results suggest that in rat stomach, both circular and longitudinal muscle layers are innervated with nitroxergic nerves which relax the gastric smooth muscle via NO-cGMP pathway.

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Sensing the Stress: the Role of the Stress-activated p38/Hog1 MAPK Signalling Pathway in Human Pathogenic Fungus Cryptococcus neoformans

  • Bahn, Yong-Sun;Heitman, Joseph
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2007.05a
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    • pp.120-122
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    • 2007
  • All living organisms use numerous signal-transduction pathways to sense and respond to their environments and thereby survive and proliferate in a range of biological niches. Molecular dissection of these signalling networks has increased our understanding of these communication processes and provides a platform for therapeutic intervention when these pathways malfunction in disease states, including infection. Owing to the expanding availability of sequenced genomes, a wealth of genetic and molecular tools and the conservation of signalling networks, members of the fungal kingdom serve as excellent model systems for more complex, multicellular organisms. Here, we employed Cryptococcus neoformans as a model system to understand how fungal-signalling circuits operate at the molecular level to sense and respond to a plethora of environmental stresses, including osmoticshock, UV, high temperature, oxidative stress and toxic drugs/metabolites. The stress-activated p38/Hog1 MAPK pathway is structurally conserved in many organisms as diverse as yeast and mammals, but its regulation is uniquely specialized in a majority of clinical Cryptococcus neoformans serotype A and D strains to control differentiation and virulence factor regulation. C. neoformans Hog1 MAPK is controlled by Pbs2 MAPK kinase (MAPKK). The Pbs2-Hog1 MAPK cascade is controlled by the fungal "two-component" system that is composed of a response regulator, Ssk1, and multiple sensor kinases, including two-component.like (Tco) 1 and Tco2. Tco1 and Tco2 play shared and distinct roles in stress responses and drug sensitivity through the Hog1 MAPK system. Furthermore, each sensor kinase mediates unique cellular functions for virulence and morphological differentiation. We also identified and characterized the Ssk2 MAPKKK upstream of the MAPKK Pbs2 and the MAPK Hog1 in C. neoformans. The SSK2 gene was identified as a potential component responsible for differential Hog1 regulation between the serotype D sibling f1 strains B3501 and B3502 through comparative analysis of their meiotic map with the meiotic segregation of Hog1-dependent sensitivity to the fungicide fludioxonil. Ssk2 is the only polymorphic component in the Hog1 MAPK module, including two coding sequence changes between the SSK2 alleles in B3501 and B3502 strains. To further support this finding, the SSK2 allele exchange completely swapped Hog1-related phenotypes between B3501 and B3502 strains. In the serotype A strain H99, disruption of the SSK2 gene dramatically enhanced capsule biosynthesis and mating efficiency, similar to pbs2 and hog1 mutations. Furthermore, ssk2, pbs2, and hog1 mutants are all hypersensitive to a variety of stresses and completely resistant to fludioxonil. Taken together, these findings indicate that Ssk2 is the critical interface protein connecting the two-component system and the Pbs2-Hog1 pathway in C. neoformans.

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Study on the Effect of the Lift Column Layout Design on Structural Strength of the Passenger Boarding Bridge (리프트 컬럼 배치설계가 탑승교 구조 강성에 미치는 영향에 대한 연구)

  • Na, Won Hyun;Koo, Hwan Jun;Bin, Soo Yeol
    • Transactions of the KSME C: Technology and Education
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    • v.3 no.4
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    • pp.307-312
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    • 2015
  • The passenger boarding bridge which provides a safe and comfort passenger pathway between an airplane and airport terminal gate is one of the apron equipment. This study investigates the effect of the lift column layout design on structural strength of the passenger boarding bridge by using finite element method, comparing deflection and stress. The overlapped zone of the tunnel frame A and B occurred at the maximum stress. The results of this research show that the lift column layout design is closely the value of the maximum stress.

Aqueous Extract of Rosa rugosa Radix Dilates Vascular Smooth Muscle Via a NO-cGMP Pathway

  • Lee, Jun-Kyoung;Li, Hua Cao;Kang, Dae-Gill;Lee, Hyuck;Jang, Ji-Yeon;Lee, Ho-Sub
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.19 no.3
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    • pp.797-801
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    • 2005
  • While conducting an in vitro screening of various medicinal plant extracts, an aqueous extract of Rosa rugosa Radix (ARR) was found to exhibit a distinct vasorelaxant activity. ARR induced a concentration-dependent relaxation of the phenylephrine-precontracted aorta. This effect disappeared with the removal of functional endothelium. Pretreatment of the aortic tissues with NG-nitro-L-arginine methyl ester (L-NAME) or 1H-[1,2,4]-oxadiazole-[4,3-]-quinoxalin-1-one (ODQ) completely inhibited the relaxation induced by ARR. ARR-induced vascular relaxations were also markedly attenuated by addition of diltiazem or verapamil. However, the relaxant effect of ARR was not blocked by pretreatment with indomethacine, tetraethylammonium (TEA), glibenclamide, atropine, or propranolol. Taken together, the present study suggests that ARR dilates vascular smooth muscle via endothelium-dependent NO/cGMP signaling.

The Development of Leaf in Amaranthus retroflexus and Chenopodium album Represented by the Plastochron (Plastochron 에 의한 Amaranthus retroflexus 와 Chenopodium album 의 잎의 성장 해석 2. 잎의 경쟁적 발달)

  • Park, Bong-Kyu;Joung-Hee Kim
    • The Korean Journal of Ecology
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    • v.8 no.3
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    • pp.127-132
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    • 1985
  • In this study, the competitive development of leaf in Amaranthus and Chenopodium were invesitgated by the complemented plastochron index. The value of PI for Amaranthus was not varied with competional ratios, while for Chenopodium it was varied. Namely in Chenopodium, the plastochron 1 was increased as competitional combinations, the plastochron 2 was decreased. These results indicated Amaranthus had advantage in competition over Chenopodium. It is surmised that these results were exhibited differences in photosynthetic pathway between Amaranthus (C4 plant) and Chenopodium (C3 plant). The linear patterns were clearly demonstrated by the differences in leaf arrangement between Amaranthus (alternate type) and Chenopodium (opposite type). From these resultss, use of plastochron seems to a useful means of evaluating plat response to various environments.

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The Reaction between Organothioenamine Phosphonate and Styrene Oxide (유기 티오엔아민 포스포네이트와 산화스티렌과의 반응)

  • Keun Jae Kim;Chi Sun Han
    • Journal of the Korean Chemical Society
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    • v.20 no.4
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    • pp.295-298
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    • 1976
  • The fact that a reaction between organothioenamine phosphonate and styrene oxide produces a derivative of cyclopropane has been proved by structural identification. This suggests that an anionic oxygen atom from the ring opening of the styrene oxide by nucleophilic attack of thioenamine phosphonate links to the phosphorous atom to from a betaine as an intermediate which is followed by cleavage of the weak P-C bond. The dextrorotatory optical activity of the product showed that the reaction was under the control of steric stability of the benzyl carbon in styrene which leads to the product through a sterically stable pathway.

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D-Erythroascorbic acid regulates cyanide-resistant respiration in Candida albicans

  • Lee, Jung-Shin;Huh, Won-Ki;Kang, Sa-Ouk
    • Proceedings of the Korean Biophysical Society Conference
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    • 2001.06a
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    • pp.42-42
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    • 2001
  • Candida albicans possess cyanide-resistant respiratory pathway, which is mediated by alternative oxidase. The activity of alternative oxidase has been found to be dependent on several regulatory mechanisms. In order to investigate the influence of D-erythroascorbic acid on respiration of C. albicans, the respiratory activity of the cells was measured with oxygen monitor. ALO1 is known to encode D-arabinono-1,4-lactone oxidase that catalyses the final step of D-erythroascorbic acid biosynthesis in C. albicans.(omitted)

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Genome sequence of carotenoid producing Sphingobacteriaceae bacterium SH-48 isolated from freshwater in Korea (카로티노이드 생산 Sphingobacteriaceae SH-48 균주의 유전체 염기서열 분석)

  • Choi, Ahyoung;Chung, Eu Jin;Nam, Young Ho;Choi, Gang-Guk
    • Korean Journal of Microbiology
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    • v.53 no.4
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    • pp.347-350
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    • 2017
  • We sequenced the genome of the Sphingobacteriaceae bacterium SH-48 isolated from the Sohan stream in Republic of Korea by using a dilution-to-extinction culturing method. The sequences were assembled into a draft genome containing 5,650,162 bp with a G + C content of 35.4% and 4,856 protein-coding genes in 2 contigs. This strain contains the carotenoid biosynthesis genes crtY, crtZ, crtD, crtI, crtB, and crtH as gene clusters. This genomic information provides new insights into the carotenoid biosynthesis pathway.