• Title/Summary/Keyword: Protein Fraction

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In vitro Antioxidant and Anti-Inflammatory Activities of Ethanol Extract and Sequential Fractions of Flowers of Prunus persica in LPS-Stimulated RAW 264.7 Macrophages (복숭아꽃 에탄올 추출물과 분획물의 in vitro 항산화 효과 및 RAW 264.7 대식세포에서의 항염증 효과)

  • Kwak, Chung Shil;Choi, Hye-In
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.10
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    • pp.1439-1449
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    • 2015
  • Prunus persica Flos (PPF) were investigated for their antioxidant and anti-inflammatory activities to find a natural functional food resource preventing degenerative diseases associated with excessive oxidative stress and chronic inflammation. PPF was extracted using ethanol (EtOH) and then sequentially fractioned by hexane (Hx), dichloromethane (DM), ethyl acetate (EA), n-butanol (BtOH), and water (DW). Contents of total phenolics and flavonoids, as well as 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging activities were measured. Anti-inflammatory effects in terms of nitric oxide (NO), prostaglandin (PG) E2, and pro-inflammatory cytokines such as interleukin (IL)-6 and tumor necrosis factor (TNF)-${\alpha}$ production were also measured using LPS-treated RAW 264.7 macrophages. EtOH extract showed relatively high antioxidant activity with high total phenolic (78.1 mg tannic acid/g) and flavonoid contents (55.3 mg rutin/g). EA fraction contained the highest total phenolic and flavonoid contents (394.6 mg tannic acid/g, 253.7 mg rutin/g), followed by BtOH (128.3 mg tannic acid/g, 93.1 mg rutin/g). EA and BtOH fractions and EtOH extract showed higher DPPH radical and ABTS radical scavenging activities than the others (P<0.05). In LPS-treated RAW 264.7 macrophages, EtOH extract ($200{\mu}g/mL$) showed significantly reduced (P<0.05) NO, PGE2, and TNF-${\alpha}$ production levels to 38.5%, 32.3%, and 48.9% of the control, respectively, as well as reduced iNOS and COX-2 protein expression. DM fraction ($50{\mu}g/mL$) showed significantly reduced (P<0.05) NO, PGE2, IL-6, and TNF-${\alpha}$ production levels to 43.5%, 13.3%, 38.7%, and 41.3% of the control, respectively, and EA fraction ($50{\mu}g/mL$) showed significantly reduced NO, PGE2, IL-6, and TNF-${\alpha}$ production levels to 44.8%, 22.4%, 45.7%, and 62.0% of the control, respectively. Taken together, EtOH extract of PPF showed potent antioxidant and anti-inflammatory activities, and EA and BtOH fractions showed comparatively stronger antioxidant activities while DM and EA fractions showed stronger anti-inflammatory activities. It can be concluded that EtOH extract of PPF and its fractions are good candidates as natural resources for the development of anti-oxidative and anti-inflammatory functional food products.

Studies on Salient Metabolites of Plant Tissues (I) -Nitrogen Metabolism and Proline Accumulation in Halophytes- (식물조직계(植物組織系)의 유효성분(有效成分)에 관(關)한 연구(硏究) [1] -내염성(耐鹽性) 식물(植物)의 Proline축적(蓄積) 및 질소대사(窒素代謝)-)

  • Cho, I.H.
    • Applied Biological Chemistry
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    • v.20 no.2
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    • pp.221-227
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    • 1977
  • Contents of proline and chloride in halophytes were $80-1700\;{\mu}g/gfw$ and 0.13-0.45 mM/gfw respectively. The content of proline was inversely proportional to that of chloride. Rhizomes of Phargmites communis Trin, a halophyte, were grown in non-saline medium and then taken to saline treatment for one or two weeks. Growth of P. communis was inhibited when salinized with 0.25M NaCl. Total nitrogen decreased and alcohol soluble nitrogen and proline increased when growth was retarded. The quantity of Fraction 1 protein decreased at 0.25M NaCl treatment. The accumulation of proline at high concentration in P. communis suggested that it might play a role in osmotic adjustment.

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Photoalteration in Biodegradability and Chemical Compositions of Algae- derived Dissolved Organic Matter (자외선에 의한 조류기원 용존유기물의 생분해도 및 화학조성변환.)

  • Imai, Akio;Matsushige, Kazuo;Nagai, Takashi;Kim, Yong-Hwan;Kim, Bom-Chul;Choi, Kwang-Soon
    • Korean Journal of Ecology and Environment
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    • v.36 no.3 s.104
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    • pp.235-241
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    • 2003
  • The effect of ultraviolet (UV) radiation on the characteristics of algae-derived dissolved organic matter (DOM) was examined by comparing the biodegradability and DOM fraction distribution of algal DOM before and after UV exposure. Algal DOM from two axenic cultures of Microcystis aeruginosa and Oscillatoria agardhii were irradiated for 24 h at a UV intensity of 42 W/$m^2$. A complete degradation of algal DOM during the UV exposure did not occur, remaining at constant concentrations of dissolved organic carbon(DOC). After UV exposure, however, microbial degradations were reduced by 17% in M. aeruginosa and 53% in O. agardhii, respectively, and decomposition rates also were two times lower in UV exposed algal DOM. In addition, the chemical compositions of algal DOM altered substantially after UV radiation exposure. The proportions of hydrophilic bases (HiB; protein-like DOM) decreased considerably in both algal DOM sources after UV exposure (16.8% and 20.0% of DOM, respectively), whereas those of hydrophilic acids (HiA; carboxylic acids-like DOM) increased as much as the decrease of the HiB fraction. Capillary ion electrophoresis (CE) analysis showed that several carboxylic acids increased significantly after UV exposure, further confirming an increase in HiA fractions. The results of this study clearly indicate that algal DOM can be changed in its chemical composition as well as biodegradability without complete degradation by UV radiation.

A Novel Glycine-Rich Region in Sox4 is a Target for the Proteolytic Cleavage in E. coli (전사활성 인자인 Sox4의 단백질 분해효소에 의한 표적 부위에 관한 연구)

  • 허은혜;최주연;장경희;김인경;임향숙
    • Korean Journal of Microbiology
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    • v.38 no.3
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    • pp.153-161
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    • 2002
  • Sox4, a transcription factor, consists of three functional domains: an HMG-box domain as a DNA binding domain, serine rich region as a transactivation domain and glycine rich region (GRR), an unknown functional domain. Although Sox4 is known to be functionally involved in heart, B-cell and reproductive system development, its physiological function remains to be elucidated. We used pGEX expression system to develop a simple and rapid method for purifying Sox4 protein in suitable forms for biochemical studies of their functions. Unexpectedly, we observed that full-length Sox4 appears to be protease-sensitive during expression and purification in E. coli. To map the protease-sensitive site in Sox4, we generated various constructs with each of functional domains of Sox4 and purified as the GST-Sox4 fusion proteins using glutathione beads. We found that the specific cleavage site for the proteolytic enzyme, which exists in E. coli, is localized within the novel GRR of Sox4. Our study suggest that the GRR of Sox4 may a target for the cellular protease action and this cleavage in the GRR may be involved in regulating physiological function of Sox4. Additionally, our study may provide a useful method for investigating the proteolytic cleavage of the target molecule in E. coli.

Diurnal Patterns in the Flow of Escapable Soluble Non-Ammonia Nitrogen Fractions in Omasal Digesta as Influenced by Barley and Rapeseed Meal Supplementation in Cows Fed Grass Silage Based Diet (목초 사일리지 급여 시 보리와 채종박 보충급여에 의한 제 3위 소화액내 Soluble Non-ammonia Nitrogen Fraction의 Flow 패턴 변화)

  • Choi, C.W.
    • Journal of Animal Science and Technology
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    • v.49 no.3
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    • pp.341-350
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    • 2007
  • The present study was conducted to measure diurnal patterns in the flow of soluble non-ammonia nitrogen (SNAN) fractions in the liquid phase of digesta entering the omasum of cows fed grass-red clover silage supplemented with barley and rapeseed meal. Four ruminally cannulated cows were fed, in a 4×4 Latin square design, grass-red clover silage alone (GS) or supplemented with (on a DM basis) 6.0kg/d of barley grain, 2.1kg/d of rapeseed meal or 6.0kg/d of barley and 2.1kg/d rapeseed meal. Omasal digesta was taken using an omasal sampling system at 1.5h intervals during a 12h feeding cycle, and SNAN fractions (free AA, peptide and soluble protein) in the omasal digesta were assessed using ninhydrin assay. Dietary supplementation numerically increased the mean flow of SNAN fractions relative to GS diet despite the lack of statistical significance. Diurnal patterns in the flow of peptide entering the omasum during a 12h feeding cycle appeared to be highest immediately after feeding, declined by 10.0h post-feeding and slightly increased thereafter. In SNAN fractions, the flow of peptide was higher for supplemented diets than for GS diet throughout the feeding cycle. Based on the microbial contribution to total SNAN using 15N, diurnal patterns in the flow of dietary SNAN for dietary supplemented diets appeared to be higher compared with GS diets. Present results may conclude that peptide flow is quantitatively the most important N in SNAN fractions and that dietary supplementation can increase peptide flow entering the omasal canal.

Cytotoxic and Apoptotic Effects of Saponins from Akebia quinata on HepG2 Hepatocarcinoma Cells (으름유래 사포닌의 HepG2 간암세포에 대한 세포독성 및 세포자살유도 효과)

  • Kang, Hye-Sook;Kang, Jae-Seon;Jeong, Woo-Sik
    • Food Science and Preservation
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    • v.17 no.3
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    • pp.311-319
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    • 2010
  • Four saponins (1~4) were isolated from Akebia quinata pericarp through bioassay-guided fractionation. Pericarps of A. quinata were extracted with ethanol and sequentially fractionated with dichloromethane, ethyl acetate, butanol and water. Compounds 1~4 from the butanol fraction were identified as 3-O-${\alpha}$-L-arabinopyranosyl hederagenin (${\delta}$-hederin), 3-O-${\alpha}$-L-rhamnopyranosyl (1${\rightarrow}$2) ${\alpha}$-L-arabinopyranoly oleanolic acid (${\beta}$-hederin), 3-O-${\beta}$-D-xylopyranosyl (1${\rightarrow}$3) ${\alpha}$-L-arabinopyranosyl hederagenin (saponin C), and 3-O ${\alpha}$-L-rhamnopyranosyl (1${\rightarrow}$2) ${\alpha}$-L-arabinopyranosyl hederagenin (${\alpha}$-hederin) based on the spectroscopic evidences, respectively. Oleanolic acid and hederagenin were identified as the corresponding sapogenins by acid-hydrolysis. These compounds exhibited strong cytotoxic activity in MTS [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxy-methoxyphenyl)-2-(4-sulfophenyl)-2H- tetrazolium, inner salt] assay on HepG2 cells. ${\beta}$-Hederin obviously attenuated the expression of bcl-2, an anti-apoptotic protein. All of the compounds also induced the activity of caspase-3, an apoptotic enzyme, while ${\alpha}$-hederin was the most potent activator of the enzyme. Our data demonstrate for the first time the apoptosis-inducing activity of A. quinata. These results suggest that A. quinata could be used as a potential source of natural cancer chemopreventive agents.

Effects of Inhalable Microparticles of Socheongryong-tang on Chronic Obstructive Pulmonary Disease in a Mouse Model (COPD 동물 모델에서 소청룡탕 흡입제형의 효과)

  • Lee, Eung-Seok;Han, Jong-Min;Kim, Min-Hee;Namgung, Uk;Yeo, Yoon;Park, Yang-Chun
    • The Journal of Korean Medicine
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    • v.34 no.3
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    • pp.54-68
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    • 2013
  • Objectives: This study aimed to evaluate the effects of microparticles of Socheongryong-tang (SCRT) on chronic obstructive pulmonary disease (COPD) in a mouse model. Methods: The inhalable microparticles containing SCRT were produced by spray-drying with leucine as an excipient, and evaluated with respect to the aerodynamic properties of the powder by Andersen cascade impactor (ACI). Its equivalence to SCRT extract was evaluated using lipopolysaccharide (LPS) and a cigarette-smoking (CS)-induced murine COPD model. Results: SCRT microparticles provided desirable aerodynamic properties (fine particle fraction of $49.6{\pm}5.5%$ and mass median aerodynamic diameter of $4.8{\pm}0.3{\mu}m$). SCRT microparticles did not show mortality or clinical signs over 14 days. Also there were no significant differences in body weight, organ weights or serum chemical parameters between SCRT microparticle-treated and non-treated groups. After 14 days the platelet count significantly increased compared with the non-treated group, but the values were within the normal range. Inhalation of SCRT microparticles decreased the rate of neutrophils in blood, granulocytes in peripheral blood mononuclear cells (PBMC) and bronchoalveolar lavage fluid (BALF) and level of TNF-${\alpha}$ and IL-6 in BALF on COPD mouse model induced by LPS plus CS. This effect was verified by histological findings including immunofluorescence staining of elastin, collagen, and caspase 3 protein in lung tissue. Conclusions: These data demonstrate that SCRT microparticles are equivalent to SCRT extract in pharmaceutical properties for COPD. This study suggests that SCRT microparticles would be a potential agent of inhalation therapy for the treatment of COPD.

Estimation of Nutritive Value of Whole Crop Rice Silage and Its Effect on Milk Production Performance by Dairy Cows

  • Islam, M.R.;Ishida, M.;Ando, S.;Nishida, T.;Yoshida, N.
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.10
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    • pp.1383-1389
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    • 2004
  • The nutritive value and utilization of whole crop rice silage (WCRS), Hamasari, at yellow mature stage was determined by three studies. In first study, chemical composition, in vivo digestibility and metabolizable energy (ME) content of WCRS was determined by Holstein steers. WCRS contains 6.23% CP, its digestibility is 48.4% and estimated TDN is 56.4%. Its ME content was 1.91 Mcal/kg DM. Gross energy (GE) retention (% of GE intake) in steers is only 22.7% most of which was lost through feces (44.7% of GE intake). It takes 81 minutes to chew a kg of WCRS by steers. In another study, the effect of Hamasari at yellow mature stage at three stages of lactation (early, mid and late lactation) and two levels of concentrate (40 or 60%) on voluntary intake, ME content and ME intake, milk yield and composition using lactating Holstein dairy cows were investigated. Total intake increased with the concentrate level in early and mid lactation, but was similar irrespective of concentrate level in late lactation. WCRS intake was higher with 40% concentrate level than with 60% concentrate. ME intake by cows increased with the concentrate level and WCRS in early lactating cows with 40% concentrate can support only 90% of the ME requirement. Milk production in accordance with ME intake increased with the increase in concentrate level in early and mid lactating cows but was similar in late lactating cows irrespective of concentrate level. Fat and protein percent of milk in mid and late lactating cows were higher with for 60% concentrate than 40%, but reverse was in early lactating cows. Solids-not-fat was higher with for 60% concentrate than 40% concentrate. Finally in situ degradability of botanical fractions such as leaf, stem, head and whole WCRS, Hamasari at yellow mature stage was incubated from 0 to 96 h in Holstein steers to determine DM and N degradability characteristics of botanical fractions and whole WCRS. Both DM and N solubility, rate of degradation and effective degradability of leaf of silage was lower, but slowly degradable fraction was higher compared to stem and head. Solubility of DM and N of stem was higher than other fractions. The 48 h degradability, effective degradability and rate of degradation of leaf were always lower than stem or head. In conclusion, voluntary intake of silage ranged from 5 to 12 kg/d and was higher with low levels of concentrate, but milk yield was higher with high levels of concentrate. Fat corrected milk yield ranged from 19 to 37 kg per day. For consistency of milk, early lactating cows should not be allowed more than 40% whole crop rice silage in the diet, but late lactating cows may be allowed 60% whole crop rice silage.

Characterization of the Water Soluble Organic Fraction Extracted from a Sewage Sludge Amended Soil (Sewage Sludge를 시용(施用)한 토양(土壤)에서 추출(抽出)한 수용성유기물(水溶性有機物)의 화학구조적(化學構造的) 특성(特性))

  • Lim, Hyungsik;Volk, V.V.;Baham, J.
    • Korean Journal of Soil Science and Fertilizer
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    • v.18 no.1
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    • pp.38-49
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    • 1985
  • The water soluble organic fractions (WSOF) from an agricultural soil (W), a soil treated with sludge for 6 years ($WS_6$), a sludge-soil mixture incubated for one week ($WS_1$), and sewage sludge (SS) were extracted, purified, and characterized by elemental analysis, functional group determinations, infrared, UV-visible, and proton nuclear magnetic resonance spectrosocpy. The SS was characterized by higher organic H, N, and P contents, a higher H/C ratio, and a lower C/N ratio than W. Total acidity carboxyl and phenolic hydroxyl group contents were generally highest in SS, intermediate in $WS_6$ and $WS_1$, and lowest in W. Overall aromatic character and aromatic carboxyl group contents were highest in W, and lowest in SS. Aliphatic proton, aliphatic carboxyl, and phenolic hydroxyl group contents were highest in SS, and lowest in W. Protein decomposition products were the pronounced components in SS, and decreased in concentration as the sludge component in the mixtures decreased. The $^1H$-NMR spectra suggested that the SS-protons were bound to a wider range of functional groups than W-protons. Structural complexities around the aromatic protons followed the following order: SS>$WS_1$>$WS_6$>W.

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Effect of irradiation on cell cycle, cell death and expression of its related proteins in normal human oral keratinocytes (방사선조사가 사람 정상 구강각화 세포의 세포주기, 세포사 및 수종 단백질의 발현에 미치는 영향)

  • Kang Mi-Ae;Heo Min-Suk;Lee Sam-Sun;Oh Sung-Ock;Lee Sul-Mi;Jeon In-Seong;Choi Soon-Chul;Park Tae-Won
    • Imaging Science in Dentistry
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    • v.33 no.3
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    • pp.143-149
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    • 2003
  • Purpose: To investigate the radiosensitivity of the normal human oral keratinocytes (NHOK), and the effect of irradiation on cell cycle and protein expression. Materials and Methods: To evaluate the radiosensitivity of NHOK, the number of colonies and cells were counted after irradiation and the SF2 (survival fraction at 2Gy) value, and the cell survival curve fitted on a linear-quadratic model were obtained. LDH analysis was carried out to evaluate the necrosis of NHOK at 1, 2, 3, and 4 days after 2, 10, and 20 Gy irradiation. Cell cycle arrest and the induction of apoptosis were analyzed using flow cytometry at 1, 2, 3, and 4 days after 2, 10, and 200y irradiation. Finally, proteins related cell cycle arrest and apoptosis were analysed by Western blot. Results: The number of survived cell was significantly decreased in a dose-dependent manner. The cell survival curve showed SF2, α, and β values to be 0.568, 0.209, and 0.020 respectively. At 200y irradiated cells showed higher optical density than the control group. After irradiation, apoptosis was not observed but G2 arrest was observed in the NHOK cells. 1 day after 10 Gy irradiation, the expression of p53 remained unchanged, the p2l/sup WAF1/Cipl/ increased and the mdm2 decreased. The expression of bax, bcl-2, cyclin B1, and cyclin D remained unchanged. Conclusion: These results indicate that NHOK responds to irradiation by G2 arrest, which is possibly mediated by the expression of p21/sup WAFl/Cipl/, and that cell necrosis occurs by high dose irradiation.

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