• Title/Summary/Keyword: Triton X

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Lanthanum-induced Inhibitions of Microsomal $H^+-ATPase$ in the Roots of Tomato ($La^{3+}$에 의한 토마토 뿌리조직 마이크로솜 $H^+-ATPase$ 활성저해)

  • Cho, Kwang-Hyun;Kim, Young-Kee
    • Applied Biological Chemistry
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    • v.46 no.2
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    • pp.84-89
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    • 2003
  • In order to find a chemical agent which is able to modulate the activity of $H^+-ATPase$, microsomal preparation was obtained from the root tissue of tomato plant and the effect of $La^{3+}$ was measured. The activities of plasma and vacuolar membrane $H^+-ATPase$ were analyzed by the inhibited activities using their specific inhibitors, vanadate and $NO_3-$, respectively. $La^{3+}$ inhibited microsomal ATPases in a dose-dependent manner and the inhibitory effect of $La^{3+}$ was suppressed by both vanadate and $NO_3-$, implying that $La^{3+}$ inhibits both plasma and vacuolar membrane $H^+-ATPase$. The Ki. values of $La^{3+}$which inhibit 50% of the activities of plasma and vacuolar membrane $H^+-ATPase$ were 57 and $78\;{\mu}M$, respectively. The $H^+-ATPase$ of the leaky microsomes made by the treatment of Triton X-100 were also inhibited by $La^{3+}$, suggesting that $La^{3+}$ directly inhibits both enzymes. Meanwhile, the inhibitory effect of $La^{3+}$ was decreased by increasing the concentration of ATP, The effect of ATP was also concentration-dependent and 7 mM ATP completely removed the inhibitory effect of $La^{3+}$. These results imply that $La^{3+}$ inhibits both plasma and vacuolar membrane $H^+-ATPases$ by decreasing the binding affinity of ATP and $La^{3+}$ can be used to control the activity or root $H^+-ATPases$.

Isolation of Cadmium-Tolerant Bacteria and Characterization of Cadmium Accumulation into the Bacteria Cell (카드뮴 내성균(耐性菌)의 분리(分離), 동정(同定)및 균체내(菌體內) 카드뮴 축적(蓄積) 특성(特性))

  • Cho, Ju-Sik;Han, Mun-Gyu;Heo, Jong-Soo
    • Korean Journal of Environmental Agriculture
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    • v.11 no.1
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    • pp.77-85
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    • 1992
  • Of the cadmium-tolerant 162 bacterial strains isolated from soils, river waters or active sludges of waste-water disposal plants in the Gyeongnam province a strain C1, which showed considerably higher growth rate in the agar plate containing 2000 ppm than any other strains isolated, was identified as a Pseudomonas putida or its similar strain when analyzed by taxonomical characteristics. Optimum pH and temperature for the growth of the P, putida were 7.0 and $30^{\circ}C$, respectively. This strain was resistant to antibiotics(ampicillin, chloramphenicol and streptomycin), and heavy metals(lithium, cupper, lead and zinc). This strain utilized salicylate, naphthalene or xylene as a sole carbon source. The rate of cadmium accumulation in P. putida cell was enhanced at low concentration of Cd in the growth media. The maximum cadmium absorption by this strain grown in 1 and l0ppm of Cd was respectively 78% and 60% 24 hrs after culture, but in 100 ppm Cd, 40% 48 hrs after culture. Addition of a non-ionic surfactant Triton X-100(0.1%) to the medium enhanced the accumulation of cadmium in the P. putida up to approximately 37%.

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Solubilization and Reconstitution of ${\Delta}^{5}-3{\beta}$-Hydroxy Steroid Acyl Transferase from the Rat Brain (쥐 뇌의 ${\Delta}^{5}-3{\beta}$-Hydroxy Steroid Acyl 전이효소의 용해 및 Liposome에서의 재구성)

  • Ko, Kyu-Jung;Park, In-Ho;Han, Beom-Ku;Jo, Do-Hyun
    • Applied Biological Chemistry
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    • v.38 no.4
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    • pp.320-324
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    • 1995
  • Solubilization of microsomal ${\Delta}^{5}-3{\beta}$-hydroxy steroid acyl transfearse(${\Delta}^{5}-3{\beta}$-OH-SAT) of rat brain and its reconstitution into liposomes were investigated. Among the detergents utilized for the solubilization, deoxycholic acid was superior to Tween 80 or Triton X-100 for the reconstituted activity of ${\Delta}^{5}-3{\beta}$-OH-SAT. The enzyme activity was shown to be affected by the nature of phospholipids used for the preparation of the liposome. Phosphatidylcholines from egg yolk and soybean showed the highest activity of ${\Delta}^{5}-3{\beta}$-OH-SAT and phosphatidylethanolamine came next. However phosphatidylserine and phosphatidic acid showed a lower activity than those obtained before the reconstitution. This study suggests that the presence of quaternary ammonium salt or amine group in the phospholipids stimulates the activity of ${\Delta}^{5}-3{\beta}$-OH-SAT. However the presence of a carboxylic group or the absence of the amine group may have an inhibitory effect on the ${\Delta}^{5}-3{\beta}$-OH SAT.

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Expression and Biochemical Characteristics of a Phospholipase D from Bacillus licheniformis (Bacillus licheniformis로부터 분리된 phospholipase D 유전자의 발현 및 생화학 특성)

  • Kang, Han-Chul;Yoon, Sang-Hong;Lee, Chang-Muk;Koo, Bon-Sung
    • Journal of Applied Biological Chemistry
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    • v.54 no.2
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    • pp.94-100
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    • 2011
  • A gene encoding a putative phospholipase D was isolated from Bacillus licheniformis and cloned into pGEM-T easy vector. The gene was expressed in E. coli BL21 (DE3) using a pET-21(a) vector containing His6 tag. Affinity purification of the recombinant phospholipase D with nickel-nitrilotriacetic acid (Ni-NTA) resin resulted major one-band by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis. The purified enzyme showed a molecular weight of 44 kDa. The optimum activity of enzyme was around pH 7.0 and the enzyme was also the most stable around this condition. The optimum temperature was about $40-45^{\circ}C$ and the enzyme still showed considerable activities at wide range of temperature. Among various detergents, Triton X-100 significantly increased the enzyme activity, resulting in 181% activity of control at 0.6 mM of the detergent. Calcium ion did not significantly affect the enzyme activity, suggesting that the enzyme might be classified into $Ca^{2+}$-independent PLD.

Studies on Protoplast Formation and Regeneration of Lyophyllum decastes (Lyophyllum decastes의 원형질체 분리와 재생에 관한 연구)

  • Bok, Jin-Woo;Kim, Jong-Pil;Jin, Mi-Rim;Choi, Eung-Chil;Kim, Byong-Kak
    • The Korean Journal of Mycology
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    • v.22 no.2
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    • pp.130-137
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    • 1994
  • This experiment was carried out to investigate proper conditions for protoplast isolation and regeneration from mycelia of Lyophyllum decastes. Novozym 234(10 mg/ml) with 0.6 M $MgSO_4$ in phosphate buffer(pH 4.0) was proper for protoplast isolation. The optimal reaction time of the mycelium with the lytic enzyme was four hours in shaking condition at 120 strokes per min. When the mycelium of L. decastes was cultured at $24^{\circ}C$ for 5 days, the formation of protoplasts was effective. The liquid medium was more effective for protoplast isolation than the solid medium. In the liquid medium, high yields of protoplasts were obtained from 0.6 M $MgSO_4$ osmotic stabilizer. Protoplasts of L. decastes were regenerated to normal hyphal growth and the regeneration frequency of the protoplasts in the complete agar medium containing Triton X-100(0.0025%) was $5.94{\sim}8.32%$. The regeneration medium stabilized with 0.6 M sucrose was the best for regeneration of the protoplasts. In contrast to protoplast formation, regeneration was inhibited by the inorganic salts used as osmotic stabilizer.

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Microbial Control of the Tobacco Cutworm, Spodopera litura (Fab.), Using S. litura Nuclea Polyhedrosis Virus. II. Formulation of S. litura Nuclear Polyhdrosis Virus as Viral Insecticides (곤충 핵다각체병바이러스를 이용한 담배거세미나방의 생물적 방제. II. 담배거세미나방 핵다각체병바이러스의 살충제 제제화)

  • 임대준;진병래;최귀문;강석권
    • Korean journal of applied entomology
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    • v.29 no.4
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    • pp.244-251
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    • 1990
  • Three viral insecticides were differently formulated with a nuclear polyhedrosis virus isolated from Spdodoptera litura by addition of feeding attractant, anti-precipitate of polyhedra, spreading agent, and UV-protectants. Sucrose was effective for attraction of larval feeding to increase the mortality and for protection of polyhedra from inactivation by sunlight when added 1% to 5% of sucrose solution to the formulations. Contents of additives to the formulations were 0.5% in polyvinyl alcohol to prohibit the precipitation of polyhedra and 0.1% in Triton X-100 to spread and wet the formulations to the plant. Inactivation of the virus under sunlight was decreased when added 800g of white carbon to 100 L of water in the white carbon formulation and 30% of molasses to the molasses's. In the formulation of white carbon and molasses mixtures, activation of the virus was increased when mixtured 500g of the former with 10% of the latter. Three formulations were persisted their pathogenicity more than 95% of mortality at 3 days p.i. Encapsulation of the polyhedral surface was more distinctively coated with the carbon and showed more effective in the residual effects of the white carbon than others, but the molasses more attractive for larval feeding.

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Enhanced Production of hCTLA4Ig through Increased Permeability in Transgenic Rice Cell Cultures (형질전환 벼 현탁세포 배양에서 투과성 증진을 통한 hCTLA4Ig의 생산성 증대)

  • Choi, Hong-Yeol;Cheon, Su-Hwan;Kwon, Jun-Young;Lim, Jung-Ae;Park, Hye-Rim;Kim, Dong-Il
    • KSBB Journal
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    • v.31 no.4
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    • pp.277-283
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    • 2016
  • In this system, rice cells were genetically modified to express human cytotoxic T-lymphocyte antigen 4-immunoglobulin (hCTLA4Ig) using RAmy3D promoter induced by sugar depletion. Even though the target protein fused with signal sequence peptide, plant cell wall can be a barrier against secretion of recombinant proteins. Therefore, hCTLA4Ig can be trapped inside cell wall or remained in intracellular space. In this study, to enhance the secretion of hCTLA4Ig from cytoplasm and cell walls into the medium, permeabilizing agents, such as dimethyl sulfoxide (DMSO), Triton X-100 and Tween 20, were applied in transgenic rice cell cultures. When 0.5% (v/v) of DMSO was added in sugar-free medium, intracellullar hCTLA4Ig was increased, on the other hand, the secreted extracellular hCTLA4Ig was lower than that of control. DMSO did not give permeable effects on transgenic rice cell cultures. And Triton X-100 was toxic to rice cells and also did not give enhancing permeability of cells. When 0.05% (v/v) Tween 20 was added in rice cell cultures, however, intracellular hCTLA4Ig was lower than that of control cultures. And the maximum 44.76 mg/L hCTLA4Ig was produced for 10 days after induction, which was 1.4-fold increase compared to that of control cultures. Especially, Tween 20 at 0.05% (v/v) showed the positive effect on the secretion of hCTLA4Ig though the decrease of intracellular hCTLA4Ig. Also, Tween 20 as a non-toxic surfactant did not affect the cell growth, cell viability and protease activity. In conclusion, secretion of hCTLA4Ig could be increased by enhancing permeability of cells regardless of the cell growth, cell viability and protease activity.

The Cleaning of Costumes of Yeosan Song's Family Excavated at Mokdal-dong in Daejeon (대전 목달동 출토 여산송씨 출토복식의 세척)

  • Baek, Young-Mee;Kwon, Young-Suk
    • Journal of Conservation Science
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    • v.25 no.2
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    • pp.219-231
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    • 2009
  • The purpose of this study is to provide basic information necessary for the cleaning of excavated costumes. For the purpose, these researchers reviewed previous records of the actual cleaning of excavated costumes and then implemented and documented the processes of cleaning the Yeosan Song's costumes excavated at Mokdal-dong, Daejeon, which could date back to the early and mid periods of Choseon Dynasty. The excavated clothes of the family provide good examples for comparing men's costume of the 15th century with men's and women's of the mid and late 16th century. The total quantity of excavated remains were 184 and textiles were cotton, silk, hemp, ramie, and union cloth. The clothing remains were processed through wet or dry cleaning in accordance with their fabric condition and the extent to which they were worn or polluted. In detail, the excavated costumes of the Yeosan Song family were cleaned in two stages. For wet cleaning, both anionic(LAS) and nonionic(Triton X-100) surfactants were respectively used as cleaning agents and for dry cleaning, a mixture of n-hexane and n-decane(the ratio of 4 to 6) and petrolic dry cleaning solvent were used. After first cleaning, some cotton, ramie and hemp which had still the stains were processed bleaching and silk which were good condition was processed dry cleaning with the organic solvent again.

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The Development of Diagnosis for Atopic Dermatitis by Evaluating the Expression of Skin Barrier Proteins Using a Non-Invasive Method (비침습적 피부장벽단백질 측정을 통한 아토피 피부염의 진단 및 유용성)

  • Kim, In Sik;Lee, Ji-Sook
    • Korean Journal of Clinical Laboratory Science
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    • v.49 no.4
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    • pp.395-400
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    • 2017
  • Atopic dermatitis is a chronic inflammatory skin disease caused by a variety of genetic and environmental factors, dysregulation of immunological response, as well as dysfunction of the skin barrier proteins. The purpose of this study is to develop an ELISA kit suitable for evaluating the expression of skin barrier proteins. Proteins were obtained from the skin via AriNo and D-Squame patches. The efficiency of protein collection from the skin, using the Arino patch, was shown to be more effective than using D-Squame; while the efficiency of lysis using 0.1% Triton-X100 was higher than that of other lysis solutions, including 0.1 M Tris-HCL, 0.1% Tween-20, and 5 mM KOH. Recombinant skin barrier proteins, such as filaggrin and involucrin, were produced by molecular biological methods. Monoclonal antibodies against filaggrin and involucrin were produced by immunization of mice, fusion of spleen cells and myeloma cells, as well as a selection of antibody-producing hybridoma cells. The filaggrin expression in the skin of subjects suffering from atopic dermatitis was lower than that in normal mice. Involucrin expression was not altered between normal individuals and subjects with atopic dermatitis. These findings contribute to an elucidation of the importance of the skin barrier protein expression in atopic dermatitis and the development of a diagnostic kit for atopic dermatitis.

Chemical Suppression of Gravitropic Bending Response in Flower Stalks of Snapdragon (Antirrhinum majus L.) (몇가지 생리활성 저해제가 금어초 절화의 굴지성 반응에 미치는 효과)

  • Kim, Yong-Sam;Kim, Dong-Hern;Hwang, Young-Soo;Jung, Jin
    • Applied Biological Chemistry
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    • v.40 no.6
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    • pp.567-571
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    • 1997
  • Numbers of chemical agents which have been shown to inhibit either auxin signal transduction pathway or ethylene formation in plant cells were applied to cut flower stems of snapdragon (Antirrhinum majus L.) and their effects on the postharvest gravitropic response were studied. The chemical treatments were done by submerging either the stem base or the top part of cut flower, which involves the gravistimulus-sensitive region, for 1 h at $25^{\circ}C$. When the chemicals were supplied from the cut stem base, the gravitropic upward bending of flower stalks kept horizontally after the treatments with 20 mM CDTA or 10 mM $CoCl_2$ was comparable to that of the untreated control, but o-vanadate showed a certain degree of effectiveness for suppressing the bending response. In contrast, the direct application of those agents to the gravitropically sensitive region of cut flowers in the presence of 0.01% Triton X-100 resulted in a substantial reduction of the gravitropic response. In the case of 20 mM $CoCl_2$ treatment, almost total elimination of gravitropism without any significant deterioration of flower quality was observed. The results indicate the possibility of preparation of a protocol involving $CoCl_2$ and a proper surfactant for commercial use to suppress the gravitropic response of cut flowers during postharvest storage and transportation.

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