• Title/Summary/Keyword: S100B protein

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Expression and Purification of Transmembrane Protein MerE from Mercury-Resistant Bacillus cereus

  • Amin, Aatif;Sarwar, Arslan;Saleem, Mushtaq A.;Latif, Zakia;Opella, Stanley J.
    • Journal of Microbiology and Biotechnology
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    • v.29 no.2
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    • pp.274-282
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    • 2019
  • Mercury-resistant ($Hg^R$) bacteria were isolated from heavy metal polluted wastewater and soil collected near to tanneries of district Kasur, Pakistan. Bacterial isolates AZ-1, AZ-2 and AZ-3 showed resistance up to $40{\mu}g/ml$ against mercuric chloride ($HgCl_2$). 16S rDNA ribotyping and phylogenetic analysis were performed for the characterization of selected isolates as Bacillus sp. AZ-1 (KT270477), Bacillus cereus AZ-2 (KT270478) and Bacillus cereus AZ-3 (KT270479). Phylogenetic relationship on the basis of merA nucleotide sequence confirmed 51-100% homology with the corresponding region of the merA gene of already reported mercury-resistant Gram-positive bacteria. The merE gene involved in the transportation of elemental mercury ($Hg^0$) via cell membrane was cloned for the first time into pHLV vector and transformed in overexpressed C43(DE3) E. coli cells. The recombinant plasmid (pHLMerE) was expressed and the native MerE protein was obtained after thrombin cleavage by size exclusion chromatography (SEC). The purification of fusion/recombinant and native protein MerE by Ni-NTA column, dialysis and fast protein liquid chromatography (FPLC/SEC) involved unfolding/refolding techniques. A small-scale reservoir of wastewater containing $30{\mu}g/ml$ of $HgCl_2$ was designed to check the detoxification ability of selected strains. It resulted in 83% detoxification of mercury by B. cereus AZ-2 and B. cereus AZ-3, and 76% detoxification by Bacillus sp. AZ-1 respectively (p < 0.05).

Neuroinflammation and Psychiatric Illness (신경염증과 정신질환)

  • Song, Hoo Rim;Lee, Hwa-Young;Shim, Se-Hoon;Kwon, Young-Joon
    • Korean Journal of Biological Psychiatry
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    • v.23 no.1
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    • pp.12-17
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    • 2016
  • Neuroinflammation is one of important allostatic loads contributory to the various psychiatric illness. It is mediated mainly by glial cells, which produce both proinflammatory and antiinflammatory cytokines, and the balance of them determines the inflammatory process in the central nervous system. S100 calcium-binding protein B, which is used as an inflammatory marker is also released by glial cells. In the molecular level, oxidative stress contributes to the neuroinflammation. Their disturbances have been revealed in the psychiatric illness and related with the dysregulation of the glutamatergic and monoaminergic systems. There is a possibility to use them as disease markers. The approach for inflammation using antiinflammatory drugs and antioxidants could be connected to the development of disease-modifying treatments. Also, a searching examination about specific subtypes who are vulnerable to inflammation in the patients is required to confirm their efficacy clearly.

Purification and Characterization of an Extracellular Xylanase of Bacillus stearothermophilus (Bacillus stearothermophilus 가 생산하는 Xylanase의 정제 및 특성)

  • 배성호;최용진
    • Microbiology and Biotechnology Letters
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    • v.19 no.6
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    • pp.592-597
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    • 1991
  • An extracellular xylanase of Bacillus stearothemophilus was purified to a single protein through a sequency of operations including ammonium sulfate fractionation, DEAE Sepharose CL-6B ion exchange chromatography, Sephadex G-100 gel filtration and heat treatment. The purified enzyme had a moleular weight of 170, 000. the pH and temperature optima for the enzyme activity were pH 9.0 and $55^{\circ}C$, respectively. The activity was enhanced by $co^{2+} \; and\; Mn^{2+}$, and inhibited by $Hg^{2+}$. Pattern of hydrolysis demonstrated that the xylanase was an endo-splitting enzyme able to break down larchwood xylan at random giving xylobiose and xylotriose as the main end products.

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Effect of Snail(Fruticiola sieboldiana) Extract on Reactive Oxygen Species(ROS) in Old Female Rats (고령 암컷 흰쥐에서 달팽이 추출물이 활성산소종에 미치는 영향)

  • Sohn, Kieho;Kim, Taehee
    • Korean Journal of Pharmacognosy
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    • v.48 no.4
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    • pp.289-297
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    • 2017
  • This study was performed to investigate the effect of 8 weeks administration of snail extract on free radical formation in old female rats (27 weeks). Rats administrated orally with snail extract at the dose of 100 mg/kg, 200 mg/kg, chondroitin sulfate 10 mg/kg and 0.9% saline (control) for 8 weeks. Hematologic profiles and hepatic enzymes were all normal. Results of analysis on snail extract were naicin, Na, protein, sugar, beta-carotene, vitamin (A, B1, B2, B6, C, E), folic acid, phosphorus, lipid, K, cholesterol, chondroitin. Hepatic lipid peroxidase content was decreased, aldehyde oxidase was decreased, glutathione peroxidase and glutathione-S transferase were not changed, but xanthine oxidase, catalase and superoxidase activities were significantly increased in snail extract fed group (p<0.001). Therefore, as the result of this study, it could be expected that the administration of snail extract for 8weeks is the potential to be use as a hepatic anti-oxidative agent.

The Inhibitiory Effects of the Scirpi rhizoma on Melanogenesis (삼릉추출물의 멜라닌 생성 저해 효과)

  • Lee, Bum-Chun;Kim, Jin-Hwa;Sim, Gwan-Sub;Zhang, Yong-He;Pyo, Hyeong-Bae
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.31 no.4 s.54
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    • pp.305-310
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    • 2005
  • To obtain effective and safe depigmenting agents, we investigated the effects of Scirpi rhizoma, a medicine among Chinese herbs, on melanogenesis. Dried S. rhizoma was refluxed with 70% aqueous ethanol and the extract was evaporated to dryness. To determine the effects as a whitening agent, various in vitro tests were performed such as free radical scavenging activity, melanin formation assay, tyrosinase activity and expression of tyrosinase, TRP-1 and TRP-2(western blot and RT-PCR) in B16 melanoma cells. S. rhizoma showed scavenging activities of free radicals and reactive oxygen species (ROS) with the $IC_{50}\;of\;638{\mu}g/mL$ against 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical and $21.7{\mu}g/mL$ against superoxide radicals in the xanthine/xanthine oxidase system, respectively. S. rhizoma significantly inhibited melanin production in B16 melanoma cells. S. rhizoma treatment(48 h) suppressed the biosynthesis of melanin up to 27% at 100{\mu}g/mL$ and reduced tyrosinase activity up to 31% at $100{\mu}g/mL$ in B16 melanoma cells. S. rhizoma was also able to significantly inhibit tyrosinase and TRP-1 expression in protein and mRNA level. These results suggest that S. rhizoma inhibited melanin biosynthesis by regulating tyrosinase activity and expression in B16 melanoma cells. Therefore, S. rhizoma may be useful as a new antioxidant and whitening agent to inhibit melanogenesis.

Protective Effects of Diallyl Sulfide against Thioacetamide-Induced Toxicity: A Possible Role of Cytochrome P450 2E1

  • Kim, Nam Hee;Lee, Sangkyu;Kang, Mi Jeong;Jeong, Hye Gwang;Kang, Wonku;Jeong, Tae Cheon
    • Biomolecules & Therapeutics
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    • v.22 no.2
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    • pp.149-154
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    • 2014
  • Effects of diallyl sulfide (DAS) on thioacetamide-induced hepatotoxicity and immunotoxicity were investigated. When male Sprague-Dawley rats were treated orally with 100, 200 and 400 mg/kg of DAS in corn oil for three consecutive days, the activity of cytochrome P450 (CYP) 2E1-selective p-nitrophenol hydroxylase was dose-dependently suppressed. In addition, the activities of CYP 2B-selective benzyloxyresorufin O-debenzylase and pentoxyresorufin O-depentylase were significantly induced by the treatment with DAS. Western immunoblotting analyses also indicated the suppression of CYP 2E1 protein and/or the induction of CYP 2B protein by DAS. To investigate a possible role of metabolic activation by CYP enzymes in thioacetamide-induced hepatotoxicity, rats were pre-treated with 400 mg/kg of DAS for 3 days, followed by a single intraperitoneal treatment with 100 and 200 mg/kg of thioacetamide in saline for 24 hr. The activities of serum alanine aminotransferase and aspartate aminotransferase significantly elevated by thioacetamide were protected in DAS-pretreated animals. Likewise, the suppressed antibody response to sheep erythrocytes by thioacetamide was protected by DAS pretreatment in female BALB/c mice. Taken together, our present results indicated that thioacetamide might be activated to its toxic metabolite(s) by CYP 2E1, not by CYP 2B, in rats and mice.

Evaluation of Chinese Brown Rice as an Alternative Energy Source in Pig Diets

  • Piao, X.S.;Li, Defa;Han, In K.;Chen, Y.;Lee, J.H.;Wang, D.Y.;Li, J.B.;Zhang, D.F.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.1
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    • pp.89-93
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    • 2002
  • A total of six crossbred barrows ($Duroc{\times}Landrace{\times}Large$ White, $44.17{\pm}1.94kg$ BW) were housed conducted to evaluate apparent fecal digestibilities of Brown Rice (BR) as an alternative energy source in growing pigs. Pigs were housed individually on metabolism crate on the basis of body weight. Four treatments contained: 1) 100% of corn-soybean meal (C100; Control diet), 2) 75% of corn-soybean meal diet plus 25% of corn meal (C25), 3) 100% of brown rice-soybean meal diet (BR100), 4) 75% of brown rice-soybean meal diet plus 25% of brown rice meal (BR25). Brown rice has an excellent gross energy and crude protein composition compared to corn. The BR used had 3,801 kcal of gross energy/kg, 8.0% crude protein, 2.6% of ether extract, 0.035% calcium and 0.35% total phosphorus. The best digestibilities of energy (87.75%), DM (81.71%) and CP (78.57%) were observed in BR 100 group and the worst were found in Corn 25 group. The nutrient digestibility was not significantly different in most nutrients. Through this experiment, BR appeared a good alternative energy source that can replace corn yellow to 100% in growing pigs. Therefore, the price relationship between corn and BR may provide an excellent opportunity for pork producers to use BR in order to reduce feed costs provided that diet has been balanced for digestible amino acids.

Quality Characteristics of Brown Sauce Prepared with Rice Powder Beurre mani$\acute{e}$ (쌀가루 Beurre mani$\acute{e}$를 사용한 브라운 소스의 품질 특성)

  • Kim, Ki-Young;Kim, Sung-Kook;Yoo, Seung-Seok
    • Journal of the East Asian Society of Dietary Life
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    • v.19 no.2
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    • pp.247-255
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    • 2009
  • This study examined the quality characteristics of brown sauce prepared with rice powder in place of wheat flour, which is the commonly used thickener. With regard to the general components, as the level of rice powder increased and its ratio in the stock increased, moisture decreased and crude protein, lipid, and ash increased. Reducing sugar content was 4.72% in the control group, and as the mixing ratio of stock to rice powder increased from 100:5 (S1) to 100:13 (S5) in the rice powder-added groups, reducing sugar content increased from 3.94% to 4.82%, respectively. In terms of Hunter's color values, as more rice power was added to the beurre mani$\acute{e}$, L, a, and b values increased. Thus, a sauce of a light brown color was appropriately made. For consistency, as more rice powder beurre mani? was inserted into the analyzer, flow was reduced and viscosity significantly increased. The pH of the control (CS) was 4.94, and there were no significant differences between the control and the rice powder-added groups. In sensory evaluations, color presented its highest score as 6.47 when the mixing ratio of stock to beurre mani$\acute{e}$ was 100:7 (S2). S2 also received the highest score for flavor at 6.32. Viscosity was highest in the control (CS) at 6.26, and was not significantly different from the 100:7 (S2) or 100:9 (S3) groups. Finally, the S2 group had the highest scores for taste (6.21) and overall-acceptability (6.30). In correlation analyses between the sensory characteristics, overall-acceptability as well as the color, flavor, viscosity, and taste of the brown sauce all presented comparatively high positive correlations, in which overall-acceptability and color had the highest correlation.

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Studies on Soybean Protein [Part ll]-Isolation and Subunit Composition of Multiple 7S Globulins- (대두(大豆) 단백질(蛋白質)에 관(關)한 연구(硏究) 제2보[第二報]-7S Globulin중의 복합단백질(複合蛋白質)의 분리(分離) 및 그 구성(構成) Subunit에 대하여)

  • Lee, C.Y.;Kim, I.S.;Kim, S.U.
    • Applied Biological Chemistry
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    • v.20 no.1
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    • pp.26-32
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    • 1977
  • The multiple 7S globulins composed of two fractions (A and B) in the electrophoresis with Davis' method were isolated at different stages of the soybean seed development. Electrophoresis of their subunits liberated in PAWU solvent [phenol-acetic acid-water (2 : 1 : 1) solution plus 5M urea] yielded 4 major bands. Observation of both the electrophoretic bands of the multiple 7S fractions(7S-A and 7S-B) and those of their subunits was suggestive of a similarity of the subunit pattern between two 7S fractions. The two fractions in multiple 7S globulins were isolated with DEAE-Sephadex A-50 column$(2.0{\sim}100cm)$ chromatography. They were separated into 2 fractions in a linear gradient concentration of 0.28 to 0.40M NaCl with phosphate buffer (pH 7.8) containing 10mM ${\beta}-mercaptoethanol$(ME). The isolated protein was dissociated into subunits with two different solvent systems; in PAWU solvent and in Tris-HCl buffer(pH 8.0) containing 1% sodium dodecyl sulfate (SDS) and 40mM ME. The dissociated subunits were subjected to electrophoresis in PAWU-treated 7.5% acrylamide gel and in 1% SDS-treated 5.6% acrylamide gel. In PAWU gel electrophoresis, total 7S globulin was separated into 5 major bands, two of which were occupied in common by two 7S fractions(7S-A and 7S-B). In SDS gel electrophoresis, total 7S globulin was separated into 7 major bands, three of which were overlapped with the subunit of the two 7S fractions. The above results alluded us to the presence of a common and/or similar subunit between the multiple 7S globulins.

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Effects of Chromium Picolinate on In Vitro Lipogenesis and Lipolysis in Adipose Tissue and Protein Synthesis in Liver Tissue of Pigs

  • Choi, Y.J.;Kim, H.G.;Cho, J.S.;Chung, I.B.;Kim, Y.H.;Han, I.K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.11 no.4
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    • pp.428-433
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    • 1998
  • The effects of chromium picolinate supplementation in pig diet were evaluated by measuring the in vitro lipogenic and lipolytic activities in adipose tissue and the protein synthetic activity in liver acinar cell in culture. Thirty-two male and thirty-two female pigs were randomly assigned to one of four dietary groups: Control, 100 ppb, 200 ppb, and 400 ppb of Cr in the form of picolinate. The chromium picolinate supplementation (p < 0.01) increased the in vitro lipolytic activity in adipose tissue of pig, but had no effects on lipogenesis. The chromium picolinate effect was greater in female pigs than in male pigs on lipolytic activity. The results from the studies with the liver acinar cells in culture indicated that chromium picolinate supplementation increased protein synthetic activity (p < 0.05). It was observed through this experiment that chromium picolinate functions not only on fat degradation but also on retained protein synthesis.