• Title/Summary/Keyword: amino acid

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Transduced Tat-α-Synuclein Protects against Oxidative Stress In vitro and In vivo

  • Choi, Hee-Soon;Lee, Sun-Hwa;Kim, So-Young;An, Jae-Jin;Hwang, Seok-Il;Kim, Dae-Won;Yoo, Ki-Yeon;Won, Moo-Ho;Kang, Tae-Cheon;Kwon, Hyung-Joo;Kang, Jung-Hoon;Cho, Sung-Woo;Kwon, Oh-Shin;Choi, Jin-Hi;Park, Jin-Seu;Eum, Won-Sik;Choi, Soo-Young
    • BMB Reports
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    • v.39 no.3
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    • pp.253-262
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    • 2006
  • Parkinson's disease (PD) is a common neurodegenerative disorder and is characterized by the progressive loss of dopaminergic neurons in the substantia nigra. Although many studies showed that the aggregation of $\alpha$-synuclein might be involved in the pathogenesis of PD, its protective properties against oxidative stress remain to be elucidated. In this study, human wild type and mutant $\alpha$-synuclein genes were fused with a gene fragment encoding the nine amino acid trans activator of transcription (Tat) protein transduction domain of HIV-l in a bacterial expression vector to produce a genetic in-frame WT Tat-$\alpha$-synuclein (wild type) and mutant Tat-a-synucleins (mutants; A30P and A53T), respectively, and we investigated the protective effects of wild type and mutant Tat-$\alpha$-synucleins in vitro and in vivo. WT Tat-$\alpha$-synuclein rapidly transduced into an astrocyte cells and protected the cells against paraquat induced cell death. However, mutant Tat-$\alpha$-synucleins did not protect at all. In the mice models exposed to the herbicide paraquat, the WT Tat-$\alpha$-synuclein completely protected against dopaminergic neuronal cell death, whereas mutants failed in protecting against oxidative stress. We found that these protective effects were characterized by increasing the expression level of heat shock protein 70 (HSP70) in the neuronal cells and this expression level was dependent on the concentration of transduced WT Tat-$\alpha$-synuclein. These results suggest that transduced Tat-$\alpha$-synuclein might protect cell death from oxidative stress by increasing the expression level of HSP70 in vitro and in vivo and this may be of potential therapeutic benefit in the pathogenesis of PD.

DEVELOPMENT OF A BIOACTIVE CELLULOSE MEMBRANE FROM SEA SQUIRT SKIN FOR BONE REGENERATION - A PRELIMINARY RESEARCH (멍게와 미더덕 피부의 천연 셀룰로오스 각질을 이용한 골재생 효능을 가진 생활성막의 개발 - 예비 연구)

  • Kim, Soung-Min;Lee, Jong-Ho;Jo, Joung-Ae;Lee, Seung-Cheol;Lee, Suk-Keun
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.31 no.5
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    • pp.440-453
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    • 2005
  • Objectives : To develop a bioactive membrane for guided bone regeneration (GBR), the biocompatibility and bone regenerating capacity of the cellulose membrane obtained from the Ascidians squirt skin were evaluated. Materials and methods : After processing the pure cellulose membrane from the squirt skin, the morphological study, amino acid analysis and the immunoreactivity of the cellulose membrane were tested. Total eighteen male Spraque-Dawley rats (12 weeks, weighing 250 to 300g) were divided into two control (n=8) and another two experimental groups (n=10). In the first experimental group (n=5), the cellulose membrane was applicated to the 8.0 mm sized calvarial bone defect and the same sized defect was left without cellulose membrane in the first control group (n=4). In the another experimental group (n=5), the cellulose membrane was applicated to the same sized calvarial bone defect after femoral bone graft and the same sized defect with bone graft was left without cellulose membrane in the another control group (n=4). Each group was sacrificed after 6 weeks, the histological study with H&E and Masson trichrome stain was done, and immunohistochemical stainings of angiogenin and VEGF were also carried out. Results : The squirt skin cellulose showed the bio-inductive effect on the bone and mesenchymal tissues in the periosteum of rat calvarial bone. This phenomenon was found only in the inner surface of the cellulose membrane after 6 weeks contrast to the outer surface. Bone defect covered with the bioactive cellulose membrane showed significantly greater bone formation compared with control groups. Mesenchymal cells beneath the inner surface of the bioactive cellulose membrane were positive to the angiogenin and VEGF antibodies. Conclusion : We suppose that there still remains extremely little amount of peptide fragment derived from the basement membrane matrix proteins of squirt skin, which is a kind of anchoring protein composed of glycocalyx. This composition could prevent the adverse immunological hypersensitivity and also induce bioactive properties of cellulose membrane. These properties induced the effective angiogenesis with rapid osteogenesis beneath the inner surface of cellulose membrane, and so the possibilities of clinical application in dental field as a GBR material will be able to be suggested.

Production and Action of Microbial Piscicidal Substance (미생물에 의한 살어성물질의 생성 및 그 작용)

  • 도재호;서정훈
    • Microbiology and Biotechnology Letters
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    • v.6 no.1
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    • pp.41-46
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    • 1978
  • Piscicidal substance produced by Streptomyces sp. isolated from soil was toxic against various kinds of fish. After extraction with CH$Cl_3$ from the culture medium, the substance was purified by avicel column chromatography. In order to test toxicity, various kinds of fish were subjected to the acqueous solution of 100 us of the substance per liter of water. Generally, the substance was toxic to most fish, but Macropodus chinenes and Misgurnus mizolepis are resistant to the substance than Gobius similis and Pseudorasbora parva. The substance was stable at pH range, 3.0 to 7.0, but labile at alkaline pH, and to heat as well. Succinic dehydrogenase on most of tissue cell of Cyprinus carpio was inhibited by this substance strongly, but spinal cord was not inhibited. By addition of Cu and Pb salts to the culture medium, piscicidal substance producibility was activated.

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Effects of Methacrylamide Treatment on Silk Fibers III. Polymerization Behavior of Methacrylamide (견섬유에 대한 메타크릴아미드의 처리효과 III. 메타크릴아미드의 중합거동)

  • 신봉섭;남중희
    • Journal of Sericultural and Entomological Science
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    • v.34 no.2
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    • pp.32-40
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    • 1992
  • Many studies have been carried out on the graft finishing in order to improve the quality of silk fiber. Various vinyl monomers, for instance, styrene, methylmethacrylate, 2-hydroxyeth-ylmethacrylate and methacrylamide, have been used practically up to date. Among these monomers, methacrylamide has been applied as the most favourable monomer onto silk fibers in recent years. The polymerization mechanism about styrene- and methylmethacrylate-grafted silk fiber has been studied by many researchers. They proposed that free radicals were formed and vinyl monomers were polymerized in silk fibroin by graft polymerization mechanism, while active sites were varied by the types of monomer and initiator as well as by the reaction condition. In general. there is another Opinion that monomers are polymerized and impregnated in the internal side of the fiber by homopolymerization, which has not been proved experimentally yet More than 10 years have been passed since methacrylamide was applied on the silk fiber, and at the present time most finishings are being achieved by methacrylamide. However, no attention has been paid to the polymerization mechanism of the methacrylamide-treated silk fiber yeL In this paper, the treatments of methacrylamide on silk fibers were studied in aqueous solution using potassium persulfate as an initiator. The polymerization mechanism of the methacrylamide-treated silk fibers was investigated and analyzed on the basis of the results of infrared spectroscopy, amino acid analysis and scanning electron microscopy. From the results of these instrumental analyses, it can be suggested that polymerization mechanism about the methacrylamide-treated silk fibers is not performed by graft polymerization which has been accepted generally in styrene and methylmethacrylate-grafted silk fibers. The different mechanism is supposed to be due to the difference in monomer types, initiator types and treatment conditions.

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Utilization of Chitin Prepared from the Shellfish Crust 2. Effect of Chitosan Film Packing on Quality of Lightly-Salted and Dried Horse Mackerel (갑각류부산물을 이용한 키틴의 제조 및 이용에 관한 연구 2. 키토산필름포장이 반염건(半鹽乾) 전갱이의 가공 및 저장 중 품질에 미치는 효과)

  • AHN Chang-Bum;LEE Eung-Ho
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.25 no.1
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    • pp.51-57
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    • 1992
  • The preservative effect of chitosan film packing on quality of lightly-salted and dried horse mackerel was studied. In preparation of chitosan film, blue crab shell chitosan was dissolved in dilute acetic acid$(1.0\%,\;v/v)$, filtered, and spreaded on plastic plate and dried at $50\pm2^{\circ}C$. The chitosan film thus obtained was neutralized with 1.0N NaOH for 2 hrs and dried at room temperature after washing several times with distilled water. The lightly-salted and dried horse mackerel product was prepared by drying for 4 hrs at $40\pm2^{\circ}C$ in hot air dryer after packing with the chitosan film. During storage at $5.0\pm0.5^{\circ}C$, moisture content of the product was higher than that of the reference, but contents of VBN(volatile basic nitrogen) , amino nitrogen, and TMA of the product on dry basis were lower than those of the reference. Viable cell count, TBA value, and peroxide value of the product were also lower than those of the reference. Judging from the result of sensory evaluation, the chitosan film packing in the storage of lightly-salted and dried horse mackerel was remarkably elongated shelf-life of the product. From the results of chemical and sensory evaluation, it was concluded that chitosan film packing was an effective method for retaining the quality of lightly-salted and dried horse mackerel.

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Preparation of Powdered Dried Sea Mussel and Anchovy for Instant Soup (진주담치 및 마른멸치 분말수우프의 제조)

  • LEE Eung-Ho;HA Jae-Ho;CHA Yong-Jun;OH Kwang-Soo;KWON Chil-Sung
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.17 no.4
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    • pp.299-305
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    • 1984
  • As one trials to utilize sea mussel and anchovy effectively, powdered instant soups were prepared and then their quality stability were examined during storage. Powdered instant soup was made by adding $3\%$ sugar, $20\%$ table salt, $5\%$ monosodium glutamate, $0.2\%$ black pepper and garlic powder to the pulverized dried sea mussel or anchovy. Powdered instant soup products, powderd products, and dried round state sea mussel or anchovy were packed with air in laminated film bag (cellophane/polyester/aluminium foil/polyester: $20{\mu}m/15{\mu}m/7{\mu}m/20{\mu}m,\;13{\times}14cm$). The contents of amino-nitrogen and volatile basic nitrogen of these products were showed little significant variations and also water activity and color value (L, a, b) of these products were little changed during storage. Thiobarbituric acid value increased up to 30 days of storage and then decreased slightly. Comparing the quality of powdered-seasoned products with that of dried round state products, there were no significant differences in stability during storage. Judging from the experimental results, the quality of powdered instant soup of sea mussel and anchovy were stable for 100 days at room temperature($25{\pm}3^{\circ}C$).

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Molecular Cloning and High-Level Expression of Human Cytoplasmic Superoxide Dismutase Gene in Escherichia coli (사람의 세포질 Superoxide Dismutase 유전자의 클로닝과 대장균내에서의 대량발현에 관한 연구)

  • 이우길;김영호;양중익;노현모
    • Korean Journal of Microbiology
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    • v.28 no.2
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    • pp.91-97
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    • 1990
  • Complementary DNA (cDNA) coding for human cytoplasmic superoxide dismutase (SOD1) (superoxide: superoxide oxidoreductase E.C.1.15.1.1) was isolated from human liver cDNA library of $\lambda$gt11 by in situ plaque hybridization. The insery cDNA gas the 5' untranslational region (UTR) and 3'UTR of SOD1 gene. Polymerase Chain Reaction (PCR) method was used fro subcloning of SOD1 structural gene. Using synthetic sense strand primer (24mer) containing a start codon and antisense strand primer (24mer), SOD1 structural gene was selectively amplified. Amplified DNA was directly cloned into the HincII site of pUC19 plasmid. Insery cDNA was subcloned into M13 mp19 and sequenced by dideowy chain termination method with Sequenase. The nucleotide sequence of insert cDNA had an open reading frame (ORF) coding for 153 amino acid residues. The structural gene of cytoplasmic SOD was placed under the control of bacteriophage $\lambda P_{L}$ regulatory sequences, generating a highly efficient expression plasmid. The production of human SOD1 in E. coli cells was about 7% of total cellular proteins and recombinant human SOD1 possessed its own enzymatic acitivity.

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Reidentification of Comamonas sp. Strain DJ-12 and Analysis of its pcbABC2D2 Genes Responsible for Degradation of 4-Chlorobiphenyl. (Comamonas sp. Strain DJ-12 의 재동정 및 4-Chlorobiphenyl 분해유전자 pcbABC2D2 의 분석)

  • 이준훈;박동우;강철희;채종찬;이동훈;김치경
    • Korean Journal of Microbiology
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    • v.40 no.2
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    • pp.121-126
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    • 2004
  • Comamonas sp. strain DJ-12 is a 4-chlobiphenyl(4CB)-degrading bacterium that was reidentified from Pseudomonas sp. DJ-12. The genomic DNA was isolated from the strain DJ-12 and amplified by PCR with primers for cloning pcbABCD genes responsible for degradation of 4CB. The amino acid sequences deduced from the nucleotide sequences of pcbA1, pcbA2, pcbA3, pcbA4, pcbB, pcbC2, and pcbD2 genes showed 91, 87, 99, 87, 97, 90 and 87% homologies with those of Pseudomonas sp. KKS102, respectively. The pcbC1D1 genes that are involved in the degradation of (4-chloro)1,2-dihydroxybiphenyl produced from 4CB by pcbAB gene products were previously reported in the recombinant plasmid pCU1 from Pseudomonas sp. DJ-12. However, the pcbC2D2 genes in the plasmid pCT4 and pCT5 cloned from Comamonas sp. DJ-12 in this study showed 51 and 62% homologies with those of pcbC1D1 in their nucleotide sequences. The pcbC1D1 and pcbC2D2 genes were found by Southern hybridization to be located at different loci on the chromosome of DJ-12 strain. These results indicate that Comamonas sp. strain DJ-12 has two different sets of pcbCD genes responsible for deg-radation of (4-chloro)1,2-dihydroxybiphenyl.

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

  • Lipton, Stuart A.
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1996.04a
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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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Isolation and Characterization of a Basic Leucine Zipper Gene in Poplar (Populus alba × P. glandulosa) (현사시나무에서 Basic Leucine Zipper 유전자의 분리와 특성 구명)

  • Yoon, Seo-Kyung;Lee, Hyoshin;Bae, Eun-Kyung;Choi, Young-Im;Kim, Joon-Hyeok;Noh, Seol Ah
    • Journal of Korean Society of Forest Science
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    • v.103 no.2
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    • pp.189-195
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    • 2014
  • Basic leucine zipper (bZIP) protein is a regulatory transcription factor that plays crucial roles in growth, development and stress response of plant. In this study, we isolated a PagbZIP1 gene that belonged to Group SE3 of bZIP from Populus alba ${\times}$ P. glandulosa, and investigated its expressional characteristics. The PagbZIP1 is 844 base pairs long and encodes a putative 144-amino-acid protein with an expected molecular mass of 16.6 kDa. The PagbZIP1 has two conserved domains including the basic and leucine zipper portions. Southern blot analysis revealed that two copies of the gene are presented in the poplar genome. PagbZIP1 was specifically expressed in the root and suspension cells. Moreover, the expression of PagbZIP1 was induced by drought, salt, cold and ABA. Therefore, our results indicated that PagbZIP1 might be expressed in response to abiotic stress through the ABA-mediated signaling pathway in poplar.