• 제목/요약/키워드: D-Amino acid

검색결과 1,166건 처리시간 0.025초

Neuronal injury in AIDS dementia: Potential treatment with NMDA open-channel blockers and nitric oxide-related species

  • Lipton, Stuart A.
    • 한국응용약물학회:학술대회논문집
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    • 한국응용약물학회 1996년도 춘계학술대회
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    • pp.19-29
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    • 1996
  • The neurological manifestations of AIDS include dementia, encountered even in the absence of opportunistic superinfection or malignancy. The AIDS Dementia Complex appears to be associated with several neuropathological abnormalities, including astrogliosis and neuronal injury or loss. How can HIV-1 result in neuronal damage if neurons themselves are only rarely, if ever, infected by the vitus\ulcorner In vitro experiments from several different laboratiories have lent support to the existence of HIV- and immune-related toxins. In one recently defined pathway to neuronal injury, HIV-infected macrophages/microglia as well as macrophages activated by HIV-1 envelope protein gp120 appear to secrete excitants/neurotoxins. These substances may include arachidonic acid, platelet-activating factor, free radicals (NO - and O$_2$), glutamate, quinolinate, cysteine, cytokines (TNF-${\alpha}$, IL1-B, IL-6), and as yet unidentified factors emanating from stimulated macrophages and possibly reactive astrocytes. A final common pathway for newonal suscepubility appears to be operative, similar to that observed in stroke, trauma, epilepsy, and several neurodegenerative diseases, including Huntington's disease, Parkinson's disease, and amyotrophic lateral sclerosis. This mechanism involves excessive activation of N-methyl-D-aspartate (NMDA) receptor-operated channels, with resultant excessive influx of Ca$\^$2+/ leading to neuronal damage, and thus offers hope for future pharmacological intervention. This chapter reviews two clinically-tolerated NMDA antagonists, memantine and nitroglycerin; (ⅰ) Memantine is an open-channel blocker of the NMDA-associated ion channel and a close congener of the anti-viral and anti-parkinsonian drug amantadine. Memantine blocks the effects of escalating levels of excitotoxins to a greater degree than lower (piysiological) levels of these excitatory amino acids, thus sparing to some extent normal neuronal function. (ⅱ) Niuoglycerin acts at a redox modulatory site of the NMDA receptor/complex to downregulate its activity. The neuroprotective action of nitroglycerin at this site is mediated by n chemical species related to nitric oxide, but in a higher oxidation state, resulting in transfer of an NO group to a critical cysteine on the NMDA receptor. Because of the clinical safety of these drugs, they have the potential for trials in humans. As the structural basis for redox modulation is further elucidated, it may become possible to design even better redox reactive reagents of chinical value. To this end, redox modulatory sites of NMDA receptors have begun to be characterized at a molecular level using site-directed mutagenesis of recombinant subunits (NMDAR1, NMDAR2A-D). Two types of redox modulation can be distinguished. The first type gives rise to a persistent change in the functional activity of the receptor, and we have identified two cysteine residues on the NMDARI subunit (#744 and #798) that are responsible for this action. A second site, presumably also a cysteine(s) because <1 mM N-ethylmaleimide can block its effect in native neurons, underlies the other, more transient redox action. It appears to be at this, as yet unidentified, site on the NMDA receptor that the NO group acts, at least in recombinant receptors.

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A Tuber Lectin from Arisaema jacquemontii Blume with Anti-insect and Anti-proliferative Properties

  • Kaur, Manpreet;Singh, Kuljinder;Rup, Pushpinder Jai;Kamboj, Sukhdev Singh;Saxena, Ajit Kumar;Sharma, Madhunika;Bhagat, Madhulika;Sood, Sarvesh Kumar;Singh, Jatinder
    • BMB Reports
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    • 제39권4호
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    • pp.432-440
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    • 2006
  • A tuber lectin from Arisaema jacquemontii Blume belonging to family Araceae was purified by employing a single step affinity chromatography using column of asialofetuin-linked amino activated silica beads and the bound lectin was eluted with 100 mM glycine-HCl buffer pH 2.5. The purified A. jacquemontii lectin (AJL) showed a single protein band with an apparent molecular mass of 13.4 kDa when submitted to SDS-polyacrylamide gel electrophoresis under reducing as well as non-reducing conditions. The native molecular mass of AJL determined by gel filtration on a Biogel P-200 column was 52 kDa and its carbohydrate content was estimated to be 3.40%. Thus AJL is a tetrameric glycoprotein. The purified lectin agglutinated erythrocytes from rabbit but not from human. Its activity was not inhibited by any of the mono- and disaccharides tested except N-acetyl-D-lactosamine having minimal inhibitory sugar concentration (MIC) 25 mM. Among the glycoproteins tested only asialofetuin was found to be inhibitory (MIC $125\;{\mu}g/mL$). A single band was obtained in native PAGE at pH 4.5 while PAGE at pH 8.3 showed two bands. Isoelectric focusing of AJL gave multiple bands in the pI range of 4.6-5.5. When incorporated in artificial diet AJL significantly affected the development of Bactrocera cucurbitae (Coquillett) larvae indicating the possibility of using this lectin in a biotechnological strategy for insect management of cucurbits. Larvae fed on artificial diet containing sub-lethal dose of AJL showed a significant decrease in acid phosphatase and alkaline phosphatase activity while esterase activity markedly increased as compared to larvae fed on diet without lectin. Out of various human cancer cell lines employed in sulphorhodamine B (SRB) assay, this lectin was found to have appreciable inhibitory effect on the in vitro proliferation of HCT-15, HOP-62, SW-620, HT-29, IMR-32, SKOV-3, Colo-205, PC-3, HEP-2 and A-549 cancer cell lines by 82, 77, 73, 70, 41, 41, 37, 29, 21 and 21% respectively.

Characterization of a Chitinase Gene Exhibiting Antifungal Activity from a Biocontrol Bacterium Bacillus licheniformis N1

  • Lee, Kwang-Youll;Heo, Kwang-Ryool;Choi, Ki-Hyuck;Kong, Hyun-Gi;Nam, Jae-Sung;Yi, Young-Byung;Park, Seung-Hwan;Lee, Seon-Woo;Moon, Byung-Ju
    • The Plant Pathology Journal
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    • 제25권4호
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    • pp.344-351
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    • 2009
  • A biocontrol bacterium Bacillus licheniformis N1 grown in nutrient broth showed no chitinolytic activity, while its genome contains a gene which encodes a chitinase. The gene for chitinase from B. licheniformis N1 was amplified by PCR and the deduced amino acid sequence analysis revealed that the chitinase exhibited over 95% identity with chitinases from other B. licheniformis strains. Escherichia coli cells carrying the recombinant plasmid displayed chitinase activity as revealed by the formation of a clear zone on chitin containing media, indicating that the gene could be expressed in E. coli cells. Chitinase gene expression in B. licheniformis N1 was not detected by RT-PCR analysis. The protein was over-expressed in E. coli BL21 (DE3) as a glutathione S-transferase fusion protein. The protein could also be produced in B. subtilis 168 strain carrying the chitinase gene of N1 strain. The crude protein extract from E. coli BL21 carrying GST fusion protein or culture supernatant of B. subtilis carrying the chitinase gene exhibited enzyme activity by hydrolyzing chitin analogs, 4-methylumbelliferyl-$\beta$-D-N,N'-diacetylchitobioside and 4-methylumbelliferyl-$\beta$-D-N,N',N"-triacetylchitotrioside. These results indicated that even though the chitinase gene is not expressed in the N1 strain, the coding region is functional and encodes an active chitinase enzyme. Furthermore, B. subtilis 168 transformants expressing the chitinase gene exhibited antifungal activity against Fulvia fulva by suppressing spore germination. Our results suggest that the proper engineering of the expression of the indigenous chitinase gene, which will lead to its expression in the biocontrol strain B. licheniformis N1, may further enhance its biocontrol activity.

The Site-Directed A184S Mutation in the HTH Domain of the Global Regulator IrrE Enhances Deinococcus radiodurans R1 Tolerance to UV Radiation and MMC Shock

  • Zhang, Chen;Zhou, Zhengfu;Zhang, Wei;Chen, Zhen;Song, Yuan;Lu, Wei;Lin, Min;Chen, Ming
    • Journal of Microbiology and Biotechnology
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    • 제25권12호
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    • pp.2125-2134
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    • 2015
  • IrrE is a highly conserved global regulator in the Deinococcus genus and contributes to survival from high doses of UV radiation, ionizing radiation, and desiccation. Drad-IrrE and Dgob-IrrE from Deinococcus radiodurans and Deinococcus gobiensis I-0 each share 66% sequence identity. However, Dgob-IrrE showed a stronger protection phenotype against UV radiation than Drad-IrrE in the D. radiodurans irrE-deletion mutant (ΔirrE), which may be due to amino acid residues differences around the DNA-binding HTH domain. Site-directed mutagenesis was used to generate a Drad-IrrE A184S single mutant, which has been characterized and compared with the ΔirrE mutant complemented strain with Drad-irrE, designated ΔirrE-E. The effects of the A184S mutation following UV radiation and mitomycin C (MMC) shock were determined. The A184S mutant displayed significantly increased resistance to UV radiation and MMC shock. The corresponding A184 site in Dgob-IrrE was inversely mutated, generating the S131A mutant, which exhibited a loss of resistance against UV radiation, MMC shock, and desiccation. qPCR analysis revealed that critical genes in the DNA repair system, such as recA, pprA, uvrA, and ddrB, were remarkably induced after UV radiation and MMC shock in the ΔirrE-IE and A184S mutants. These data suggested that A184S improves the ability against UV radiation and MMC shock, providing new insights into the modification of IrrE. We speculated that the serine residue may determine the efficiency of DNA binding, leading to the increased expression of IrrE-dependent genes important for protection against DNA damage.

폭약 2,4,6-Trinitrotoluene에 노출된 분해세균 Stenotrophomonas sp. OK-5의 세포반응 (Cellular Responses of the TNT-degrading Bacterium, Stenotrophomonas sp. OK-5 to Explosive 2,4,6-Trinitrotoluene (TNT))

  • 장효원;송승열;김승일;강형일;오계헌*
    • 미생물학회지
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    • 제38권4호
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    • pp.247-253
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    • 2002
  • 환경오염원으로서 폭약 2,4,6-trinitrotoluene (TNT)에 대한 TNT 분해세균 Stenotrophomonas sp. OK-5의 세포반응에 대하여 조사하였다. 아치사조건의 TNT농도와 노출시간에 따른 균주 OK-5의 생존율을 분석한 결과, 이 세균의 생존율은 스트레스 충격 단백질의 생성과 비례하였다. 총세포 지방산 조성분석에서 균주 OK-5는 tryp-ticase soy agar에서 자랄 때보다 TNT 배지에서 자랄 때 여러 가지 종류의 지방산이 생성되거나 사라지는 것이 밝혀졌다. 주사전자현미경하에서 TNT에 노출된 세포는 쭈글쭈글하고 불규칙적인 간상형으로 나타났다. Anti-DnaK와 anti-GroEL을 이용하여 SDS-PAGE와 Western blot을 통한 분석으로 균주 OK-5는 70 kDa DanK와 60 kDa GroEL을 포함하는 몇가지 스트레스충격단백질을 생성하는 것으로 밝혀졌다. TNT에 노출된 OK-5 배양에서 수용성 단백질 분획에 대하여 2-D PAGE를 실시하였으며, pH 3에서 pH 10의 범위에서 약 300여 개 spot들이 silver로 염색된 gel상에서 관찰되었다. 이들 가운데 TNT의 반응으로 현저하게 유도되고 발현된 10개의 spot들을 확인하였으며, 2개의 단백질, spot #1과 spot #10에 대한 내부아미노산 서열을 ESI-Q TOF로 분석한 결과, Xylella fastidiosa의 DnaK protein XF2340와 Mesorhizobium loti의 스트레스 유도단백질로 각각 밝혀졌다.

잡초성벼의 superoxide dismutase cDNA cloning과 재배벼로의 형질전환 (Isolation of Superoxide Dismutase cDNAS from an Weedy Rice Variety and Transformation of a Cultivated Rice Variety)

  • 박상규;박종석;이승인;서석철;김병극;조윤래;서학수
    • 한국환경농학회지
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    • 제21권2호
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    • pp.156-161
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    • 2002
  • 냉해나 한발등의 환경스트레스에 대해 저항성을 유발하는 유전자를 환경스트레스에 강한 잡초성벼로부터 선발하고 이들 유전자를 재배벼에 도입하여 도입유전자 산물의 과량 발현을 통해 냉해나 한발 등에 대한 저항성이 향상된 벼를 선발하고자 하였다. 잡초성벼인 Bhutan 14Ad로부터 냉해 및 한발 저항성 유전자로 알려진 superoxide dismutase (SOD) cDNA를 분리하고자 mRNA를 분리하고 이 분리된 mRNA를 이용해 reverse transcriptase PCR방법으로 SOD cDNA를 cloning 하였다. 그 결과 2종의 SOD cDNA가 cloning되어 SOD-A, SOD-B로 명명하였다. 이들 cDNA의 염기서열을 결정한 결과 이들은 아미노산 서열 상동성이 88.4%를 나타내었으며, SOD-A는 Oryza sativa 계열의 Cu/Zn SOD유전자인 GenBank accession No. L36320와 99.3% 동일하였으며, SOD-B는 accession No. D01000과 100% 동일하였다. 이들 SOD-A와 SOD-B cDNA를 재배벼인 낙동벼에 형질전환하여 형질전환체 벼를 선발하였으며, 이들 형질전환체 벼의 냉해저항성및 한발저항성 검정을 통해 저항성이 향상된 형질전환체 벼를 선발하고 있다.

단백질 가시화 형태에 따른 정보표현적합도 평가 (Evaluation of Information Representation Goodness-of-fit According to Protein Visualization Pattern)

  • 변재희;최유주;서정근
    • 인터넷정보학회논문지
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    • 제16권2호
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    • pp.117-125
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    • 2015
  • 단백질 기능을 규명하는 단백질 구조 정보는 단백질 의약품의 약효를 증진시키고, 개발을 단축시키는데 큰 영향을 미친다. 따라서 단백질의 구조를 효과적으로 분석하기 위한 단백질 가시화에 대한 연구가 증가하고 있다. 하지만 단백질 가시화에 대한 연구가 단백질의 구조를 예측하거나 렌더링의 속도를 향상시키는 것을 중심으로 이뤄지고 있으며, 단백질 가시화 형태에 따른 정보 전달 효용성에 대한 연구는 미비한 실정이다. 본 연구는 단백질 의약품에 대한 효율적인 정보 서비스의 사전 연구로써 단백질 1, 2차구조 혼합가시화 형태별 정보표현적합도를 분석하였다. 단백질 1, 2차구조 혼합가시화 형태는 대표적 가시화 서비스인 Chimera, PDB, Cn3D와 기존 가시화 서비스의 문제점을 개선한 단백질 1, 2차구조 혼합가시화 형태를 대상으로 하였다. 정보표현적합도를 구하기 위한 정보요인은 피험자 분석 결과를 바탕으로 단백질 1차구조, 아미노산 위치, 단백질 2차구조, 단백질 2차구조 비율정보로 구분하였으며, 피험자는 단백질 의약품 업계종사기간이 5년 이상인 전문가 집단을 대상으로 하였다. 그 결과 단백질 1, 2차구조 혼합가시화형태별 정보표현적합도에는 유의미한 차이가 있었으며, 가시화 형태별 정보 전달 효용성에 차이가 있음을 입증할 수 있었다.

벼의 칼슘-의존성 단백질 카이네즈 유전자인 OsCPK11의 기능적 분석 (A Functional Analysis of OsCPK11, a Calcium-dependent Protein Kinase (CDPK) Gene in Rice)

  • 이수희;이정은;필립 데이;사이몬 길로이;김성하
    • 생명과학회지
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    • 제27권11호
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    • pp.1233-1244
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    • 2017
  • Calcium-dependent protein kinases (CDPKs)는 칼슘 이온을 매개로 한 신호전달 경로에서 중요한 역할을 한다. 벼(Oryza sativa)에는 29개의 CDPKs가 확인되었지만 그들의 기능은 완벽히 밝혀지지 않았다. 이 연구는 OsCPK11 유전자에 초점을 맞춰 그것의 기능적인 특징을 조사하였다. 벼의 어린 잎, 성장한 잎, 꽃에서 OsCPK11 유전자의 조직-특이적 발현이 확인되었고, 지베렐린이 처리된 벼의 호분층에서도 이 유전자의 발현을 확인할 수 있었다. Tos-17이 삽입된 oscpk11의 표현형에서 돌연변이체 각각의 키는 야생형과 구분되지 않았지만, 영과의 수나 무게는 통계적으로 유의미한 차이가 있었다. 덧붙여 많은 돌연변이체 낟알의 배젖에서 white belly materials이 확인되었다. OsCPK11의 cDNA가 cloning되었고, 약 60.5 kD인 OsCPK11 단백질이 GST affinity chromatography와 SDS-PAGE에 의해 얻어졌다. 아미노산 서열 분석을 통해 OsCPK11이 전형적인 CDPKs의 구조적 특징을 가짐을 알 수 있었다. 이 결과는 OsCPK11유전자의 기능과 식물에서 칼슘 이온을 매개로 한 신호전달 경로에 CDPK 역할의 유용한 정보를 제공해줄 것이다.

즉석라면 스-프의 감마선 살균 (Radicidation of the Condiment for Soup of Instant Noodle(Ramen))

  • 변명우;권중호;차보숙;조한옥;강세식
    • 한국식품영양과학회지
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    • 제18권1호
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    • pp.14-18
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    • 1989
  • 즉석 라면용 스-프의 살균법 개발을 위해 이들의 미생물 오염도와 방사선 조사 및 ethylene oxide훈증처리에 의한 살균효과 및 이화학적 특성 변화를 조사한 결과는 다음과 같다. 5종의 시료의 총균수는 $10^4{\sim}10^6/g$이였고 내열성 및 내산성세균이 $10^4{\sim}10^6/g$로서 총균수의 90%이상을 차지 하였다. 효모 및 곰팡이도 $10^2{\sim}10^3/g$정도 오염되어 있었다. 살균효과에 있어서 $7{\sim}10kGy$조사로 효모 빛 곰팡이는 완전 사멸되었고, 총균수는 $10^3/g$이하로 격감되었거나 사멸되었으며 이들의 $D_{10}$값은 $2.33{\sim}3.33kGy$범위였다. 이화학적 특성 변화에서 l0kGy조사구 와 ethyleneoxide처리구는 시료에 따라 차이는 있으나 산패도, 색도, 아미노태 질소는 다소 영향을 미쳤으며, ethylene oxide 처리구는 방사선 조사구에 비해 더욱 현저한 변화를 나타 내였다.

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울산 해안의 최종간빙기 최온난기 추정 해성단구 (The estimation of the marine terrace of the Last Interglacial culmination stage(MIS 5e) in the Sanhari of Ulsan coast,southeastern Korea)

  • 최성길
    • 한국지형학회지
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    • 제23권2호
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    • pp.47-59
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    • 2016
  • The formation age and depositional environment of the marine terrace I of the estimated paleoshoreline altitude of 18m in Sanhari of Ulsan coast, southeastern Korea were investigated on the basis of examination of lithofacies and stratigraphy of terrace deposits. Marine deposits of the terrace is composed of rounded boulders(70cm in diameter) and rounded pebbles(1.0cm in diameter) which overlay them. The above rounded boulders which lie on the paleo-shore platform are considered to have been formed by wave abrasion in the same period that the paleo-shore platform was developed. The rounded pebbles which lie on the rounded boulder layer are considered to have been deposited in gravel beach and berm environment, judging from the laminae developed in this layer. The paleo-shore platform and marine rounded gravel layer of the terrace are assumed to have been formed in the large transgression period of the Last Interglacial culmination stage(MIS 5e), judging from the comparision of the formation age of 125ka B.P. of Juckcheon terrace I in the adjacent Pohang coast which was dated by amino acid dating. The terrestrial deposit of this terrace was largely composed of angular and subangular gravel mixed with marine rounded pebble which has been carried away mainly from the deposit of previous marine terraces and redeposited in this terrace. The lowest peat layer of terrastrial deposit was considered to have been deposited during the period from the late MIS 5e which is the estimated finishing time of deposition of the above marine gravels to the early stage of following regression period(MIS 5d) in which the sea level was still high. The sediments of angular and subangular gravel deposit which lie on this peat layer were assumed to have been deposited during the period from the early stage of the first regression period(MIS 5d) of the Last Interglacial to the Last Glacial. The lower part of the angular gravel layer is composed of the deposits of the fluvial and colluvial sediments, whereas most of the upper and middle part of the layer is mainly composed of angular gravels of colluvial sediments formed in the cold environment.