• Title/Summary/Keyword: Binding Capacity

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Role of stearyl-coenzyme A desaturase 1 in mediating the effects of palmitic acid on endoplasmic reticulum stress, inflammation, and apoptosis in goose primary hepatocytes

  • Tang, Bincheng;Qiu, Jiamin;Hu, Shenqiang;Li, Liang;Wang, Jiwen
    • Animal Bioscience
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    • v.34 no.7
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    • pp.1210-1220
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    • 2021
  • Objective: Unlike mammals, goose fatty liver shows a strong tolerance to fatty acids without obvious injury. Stearyl-coenzyme A desaturase 1 (SCD1) serves crucial role in desaturation of saturated fatty acids (SAFs), but its role in the SAFs tolerance of goose hepatocytes has not been reported. This study was conducted to explore the role of SCD1 in regulating palmitic acid (PA) tolerance of goose primary hepatocytes. Methods: 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide was examined to reflect the effect of PA on hepatocytes viability, and quantitative polymerase chain reaction was used to detect the mRNA levels of several genes related to endoplasmic reticulum (ER) stress, inflammation, and apoptosis, and the role of SCD1 in PA tolerance of goose hepatocytes was explored using RNA interfere. Results: Our results indicated that goose hepatocytes exhibited a higher tolerant capacity to PA than human hepatic cell line (LO2 cells). In goose primary hepatocytes, the mRNA levels of fatty acid desaturation-related genes (SCD1 and fatty acid desaturase 2) and fatty acid elongate enzyme-related gene (elongase of very long chain fatty acids 6) were significantly upregulated with 0.6 mM PA treatment. However, in LO2 cells, expression of ER stress-related genes (x box-binding protein, binding immunoglobulin protein, and activating transcription factor 6), inflammatory response-related genes (interleukin-6 [IL-6], interleukin-1β [IL-1β], and interferon-γ) and apoptosis-related genes (bcl-2-associated X protein, b-cell lymphoma 2, Caspase-3, and Caspase-9) was significantly enhanced with 0.6 mM PA treatment. Additionally, small interfering RNA (siRNA) mediated downregulation of SCD1 significantly reduced the PA tolerance of goose primary hepatocytes under the treatment of 0.6 mM PA; meanwhile, the mRNA levels of inflammatory-related genes (IL-6 and IL-1β) and several key genes involved in the phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT), forkhead box O1 (FoxO1), mammalian target of rapamycin and AMPK pathways (AKT1, AKT2, FoxO1, and sirtuin 1), as well as the protein expression of cytochrome C and the apoptosis rate were upregulated. Conclusion: In conclusion, our data suggested that SCD1 was involved in enhancing the PA tolerance of goose primary hepatocytes by regulating inflammation- and apoptosis-related genes expression.

Recombinant human KAI1/CD82 attenuates M1 macrophage polarization on LPS-stimulated RAW264.7 cells via blocking TLR4/JNK/NF-κB signal pathway

  • Hyesook Lee;Jung-Hwa Han;Kangbin An;Yun Jeong Kang;Hyun Hwangbo;Ji Hye Heo;Byung Hyun Choi;Jae-Joon Kim;Seo Rin Kim;Soo Yong Lee;Jin Hur
    • BMB Reports
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    • v.56 no.6
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    • pp.359-364
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    • 2023
  • KAI1/CD82, a membrane tetraspanin protein, can prevent various cancers and retinal disorders through its anti-angiogenic and anti-metastatic capacity. However, little is known about its anti-inflammatory effect and molecular mechanism. Therefore, the present study aimed to inLPSvestigate effect of a recombinant protein of the large extracellular domain of human KAI1 (Gly 111-Leu 228, rhKAI1) on lipopolysaccharides (LPS)-stimulated RAW264.7 macrophage-like cells and mouse bone marrow-derived macrophages (BMDM) and to identify its underlying mechanism. Our data showed that rhKAI1 suppressed expression levels of classically macrophages (M1) phenotype-related surface markers F4/80+CD86+ in LPS-stimulated BMDM and RAW264.7 cells. In addition, LPS markedly increased mRNA expression and release levels of pro-inflammatory cytokines and mediators such as interleukin (IL)-1β, IL-6, tumor necrosis factor-α, cyclooxygenase-2, nitric oxide and prostaglandin E2, whereas these increases were substantially down-regulated by rhKAI1. Furthermore, LPS strongly increased expression of NF-κB p65 in the nuclei and phosphorylation of ERK, JNK, and p38 MAPK. However, nuclear translocation of NF-κB p65 and phosphorylation of JNK were greatly reversed in the presence of rhKAI1. Especially, rhKAI1 markedly suppressed expression of toll-like receptor (TLR4) and prevented binding of LPS with TLR4 through molecular docking predict analysis. Importantly, Glu 214 of rhKAI1 residue strongly interacted with Lys 360 of TLR4 residue, with a binding distance of 2.9 Å. Taken together, these findings suggest that rhKAI1 has an anti-inflammatory effect on LPS-polarized macrophages by interacting with TLR4 and down-regulating the JNK/NF-κB signaling pathway.

Natural killer cell activity in mice infected with Acanthamoeba culbertsoni (Acanthamoeba culbertsoni 감염에 있어 자연살세포의 활성)

  • Hyeon, Dong-Geun;Sin, Ju-Ok;Im, Gyeong-Il
    • Parasites, Hosts and Diseases
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    • v.30 no.2
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    • pp.101-112
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    • 1992
  • The natural killer cell activity of splenocytes and TBC, active NK cells, recycling capacity of natural killer cells were observed by means of both the 51Cr-release cytotoxicity assay and single cell cytotoxicity assay against YAC-1. CSH/HeJ mice were infected intranasally with $1{\times}10^4{\;}or{\;}1{\times}10^5$ trophosoites of pathogenic Acanthamoeba culbertsoni. The infected mice showed mortality rate of 34% in $1{\times}10^4$ group and 65% in 1{\times}10^5 group, and mean survival time was $16.40{\pm}3.50$ {\;}and{\;}3.20{\pm}4.09$ days respectively. The cytotoxic activity of natural killer cells of the 2 groups was significantly higher than that of non-infected mice from the 12th hour to the 2nd day after infection, showing the highest on the first day. On the l0th day after infection, the cytotoxic activity of natural killer cells was significantly suppressed as compared with that of the control. There was no significant difference in NK cell cytotoxicity between two infected groups. The targetbinding capacity and active NK cells of natural killer cells in $1{\times}10^5$ trophosoite infected mice was significantly increased on the 12th hour and the first day after infection as compared with the control group. Maximal recycling capacity (MRC) was not changed during the observation period. The present results indicated that the elevation of natural killer cell activity in the mice infected with A. culbertsoni was due to elevation of target.binding capacity and increased active NK cells of natural killer cells, and not due to the maximal recycling capacity of the individual NK cell, and there was no difference between two experimental dose groups.

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Lead Biosorption by Alginate Beads Immobilizing Aspergillus niger (Aspergillus niger를 고정화한 Alginate Bead에 의한 납 흡착)

  • Bang, Byung-Ho
    • Applied Biological Chemistry
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    • v.44 no.3
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    • pp.185-190
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    • 2001
  • Alginate, a well-known biopolymer, is universally applied for immobilization of microbial cells. Biosorption characteristics of lead by waste biomass of immobilized A. niger beads, used in fermentation industries to produce citric acid, were studied. The immobilized A. niger beads, prepared via capillary extrusion method using calcium chloride, were applied in the removal of lead. Pb uptake was the highest in A. niger beads cells grown for 3 days with medium producing citric acid (12% sucrose, 0.5% $NH_4NO_3$, 0.1% $KH_2PO_4$, and 0.025% $MgSO_4$). Lead uptake by the immobilized A. niger beads and free A. niger mycellia beads increased sharply with time. However, while uptake by the immobilized A. niger beads continued to increase slowly, that by free A. niger mycellia beads stopped after 30 min. The optimum pH and temperature of lead uptake were found to be 6 and $35^{\circ}C$, respectively. The maximum uptake of lead was achieved with $50{\sim}100$ beads and 50 ml lead solution in a 250-ml Erlenmeyer flask, while, at over 100 beads, uptake of the lead decreased. The order of biosorption capacity for heavy metals was Pb>Cu>Cd. Pb uptake capacity of the immobilized A. niger beads treated with 0.1 M $CaCI_2$, 0.1 M NaOH, and 0.1 M KOH decreased compared to the untreated beads. On testing the desorption of Pb from the immobilized A. niger beads, re-uptake of Pb was found possible after desorption of the binding metal with 0.1 M HCI.

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Effect of Immobilization Method in the Biosorption and Desorption of Lead by Algae, Chlorella pyrenoidosa (Chlorella pyrenoidosa에 의한 납 흡.탈착시 고정화 방법의 영향)

  • Shin, Taek-Soo;Lim, Byung-Seo;Lee, Sang-Woo;Rhu, Kwon-Gul;Jeong, Seon-Ki;Kim, Kwang-Yul
    • Journal of Korean Society of Environmental Engineers
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    • v.31 no.8
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    • pp.663-672
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    • 2009
  • In this studies, the adsorption test using Chlorella pyrenoidosa was conducted to examine the effect of Pb adsorption according to various immobilized methods such as Ca-alginate, K-carrageenan, and Polyacrylamide. From the results, the duration to need to reach adsorption equilibrium was delayed according to the immobilization. And, the higher adsorption capacity of immobilized Chlorella pyrenoidosa was represented in the higher concentration of Pb, the smaller amount of immobilizing agent, and the higher pH of solution. The maximum adsorption capacity of Pb was shown in the adsorption test using Chlorella pyrenoidosa immobilized with Ca-alginate even though it was sensitive pH. The adsorption results properly represented with Freundlich isotherm equations. And, pseudo second-order chemisorption kinetic rate equation was applicable to all the biosorption data over the entire time range. The FT-IR analysis showed that the mechanism involved in biosorption of Pb by Chlorella pyrenoidosa was mainly attributed to Pb binding of carbo-acid and amide group. Adsorbed Pb on immobilized Chlorella prenoidosa was easily desorbed in the higher concentration of desorbents(NTA, HCl, EDTA, $H_2SO_4,\;Na_2CO_3$). Among the several desorbents, NTA showed the maximum desoption capacities of Pb from Chlorella pyrenoidosa immobilized with Ca-alginate and K-carrageenan and EDTA was the most effective in Chlorella pyrenoidosa immobilized with polyacrylamide. The desoprtion efficiency in the optimum condition was 90.0, 83.0, and 80.0%, respectively.

A Study on the Role of Service Design in Creating Resident-driven Safe Community (주민주도형 안전 공동체 조성에 있어 서비스디자인의 역할 탐색)

  • Jeon, Young-Ok
    • Journal of Digital Convergence
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    • v.15 no.6
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    • pp.407-414
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    • 2017
  • With an increase in urban crimes in various forms, this study is intended to analyze the effectiveness of the service design that presented a new model resolving crime risks through differentiated thinking paradigm and problem approaches. The empirical case addressed in this study is 'the project to create resident-driven safe community in Duryu-dong, Dalseo-gu, Deagu though service design'. This project is evaluated as having prepared a prevention-oriented local safety system through a preemptive and resident-centered process. The project was promoted as a 'natural monitoring capacity building program' for residents to prevent local crimes, a 'social role expansion program' for local safety, and 'crime prevention environment design', which provides comprehensive solutions for residents' safety. Here, designers act as exerts in designing a task-based platform that can be driven by residents rather than a visual environment improver, and reorganizing the local ecosystem by expanding the opportunities for residents to interact. This case identifies the role of service design as binding the solidarity of local residents beyond the improvement of the crime environment and giving them the potential capacity to maintain a safe living space in relation to a safety issues of community.

Effect of Heat Treatments on Physical Properties and in vitro Glucose, Bile Acid, and Cadmium Transport Retardation of Wax Gourd (Benincasa hispida) (동아의 물리적 특성 및 in vitro 포도당, 담즙산, 카드뮴 투과억제 효과에 대한 열처리 영향)

  • Ju, In-Ok;Jung, Gi-Tai;Ryu, Jeong;Kim, Young-Soo
    • Korean Journal of Food Science and Technology
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    • v.35 no.6
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    • pp.1117-1123
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    • 2003
  • The effects of heat treatment on the physical and physical and physiological properties of wax gourd (Benincasa hispida) were examined. The applied heat treatments were autoclaved at $121^{\circ}C$ for 1 hr, boiled for 30 min, and microwaved at 680 W for 5 min. The water retention capacity (WRC) of the wax gourds was 9.43 g/g for the microwaved samples, 5.12 g/g for the boiled samples, 4.63 g/g for the raw samples, and 2.61 g/g for the autoclaved samples. Heat treatment caused to increase swelling by up to $4.4{\sim}7.8\;mL/g$. Calcium binding capacity of heat-treated wax gourd increased in the order of microwaved, boiled, raw, autoclaved samples. Scanning electron microscopy (SME) showed that autoclaving caused the most severe structural modifications, while microwave treatment produced the least modifications. The retarding effect on glucose and bile acid transport depended on the heat treatment. Only boiling showed the glucose retardation effect. Bile acid retardation effect increased in order of boiling (22.9%), autoclaving (17.1%), microwave treatment (14.3%), and raw wax gourd (8.6%). The cadmium retardation effect was significantly high in all samples.

Preparation and properties of gelatin from conger eel skin (붕장어껍질로부터 젤라틴의 제조 및 그 특성)

  • Ihm, Chi-Won;Kim, Poong-Ho;Kim, Jin-Soo
    • Applied Biological Chemistry
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    • v.39 no.4
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    • pp.274-281
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    • 1996
  • To prepare edible skin gelatin of conger eel such as material fur quality improvement of surimi gel, the defatted skin was limed with 1% calcium hydroxide at $5^{\circ}C$ for 2 days, washed thoroughly with tap water, extracted with 8 volumes of distilled water to dehydrated skin for 2 hours at $50^{\circ}C$. The gelatin extract was centrifuged, filtered and then passed through anion(Amberlite 200C) and cation (Amberlite IR 900) resins. The purified gelatin solution was evaporated and dried by hot-air blast$(40^{\circ}C)$. The gelatin prepared by above condition had the highest quality as revealed by physical property values i.e. 240.5 g in gel strength, $28.0^{\circ}C$ in melting point and $28.0^{\circ}C$ in gelling point. Funtional property values were 56.8% in solubility, 1.8 ml/g in oil binding capacity, 55.0% in emulsifying capacity and 48.5% in emulsifying stability. jelly strength and senso교 evaluation of surimi gel from fish with red muscle were not improved by addition of emulsifying curd from conger eel skin gelatin as emulsifier. Therefore, the conger eel skin gelatin requires a suitable modification of functional group and improvement of processing operation to utilize as a material for quality Improvement of surimi gel.

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Effect of Molokhia (Corchorus olitorius) and Its Mucilage on Cholesterol Metabolism in High Cholesterol Fed Rats (몰로키아가 고 콜레스테롤 식이를 투여한 흰쥐에서의 콜레스테롤 대사에 미치는 영향)

  • Jung, Chang-Hwa;Choi, In-Wook;Kim, Sung-Ran;Seog, Ho-Moon
    • Korean Journal of Food Science and Technology
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    • v.35 no.3
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    • pp.379-385
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    • 2003
  • Effects of molokhia (Corchorus olitorius) and its mucilage on in vitro bile acid adsorption capacity and lipid composition using cholesterol-fed SD rats were investigated. Mucilage showed stronger affinity toward bile acid than hot water extract of molokhia powder. As the extracting temperature increased from 50 to $80^{\circ}C$, bile acid binding capacity of mucilage also increased from 41 to 83%. When molokhia powder or its mucilage added into cholesterol diet at 5 or 10% levels (as fiber source) was compared with cellulose-added group, total cholesterol and triglyceride levels of plasma showed no significant differences, whereas, HDL-cholesterol level of cellulose group significantly increased. Accumulation of cholesterol and triglyceride in liver were significantly inhibited in molokhia and mucilage groups. Molokhia and mucilage lowered the liver weights significantly. As the concentration of molokhia or mucilage increased, cholesterol and triglyceride levels in the liver was lowered. Cholesterol and triglyceride excreted through feces were significant increased in molokhia- or mucilage-fed group, with excretion of cholesterol by molokhia-fed group being mere distinct.

Rapid Purification of Glucose-6-Phosphate Dehydrogenase by Affinity Chromatography (Affinity Chromatography를 이용한 Glucose-6-Phosphate Dehydrogenase의 신속한 정제방법 개발)

  • 이한수;임정빈
    • Korean Journal of Microbiology
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    • v.21 no.4
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    • pp.221-228
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    • 1983
  • An improved procedure for the rapid purification of glucose-6-phosphate dehydrogenase from extracts of Saccharomyces cerevisiae was developed by using affinity chromatography. Among six affinty media tested, $NADP^+ -agarose$ and Affi-gel Blue were more effective than others (i.e., Affi-gel Red, AMP-agarose, ATP-agarose, and $NAD^+ -agarose$). Conditions to desorb the enzyme bound to the affinity media were examined to increase the purity as well as yield. The best result was obtained when the column was developed with a linear gradient of KCl (0-1.0M). In case of Affi-gel Blue, introduction of $NAD^+$ (15mM) washing step prior to the salt gradient was most effective to remove $NAD^+ -binding$ proteins. For a large scale preparation of G-6-P dehydrogenase higher recovery was obtained by Affi-gel Blue than $NADP^+ -agarose$, however, the purity of the enzyme was decreased by 10 times if the former was used as the affinity medium. The capacity of Affi-gel Blue for G-6-P dehydrogenase was found to be 5 times higher than that of $NADP^+ -agarose$. Furthermore Affi-gel Blue could be reused repeatedly and its preparation is relatively easier and less expensive than $NADP^+ -agarose$.

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