• Title/Summary/Keyword: Protein Fraction

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Selective Extraction of Phospholipids from Soybeans with Mixture of Supercritical Carbondioxide and Ethanol (초임계 $CO_2$와 에탄올 혼합물에 의한 대두인지질의 선택적 추출)

  • Lee, Mi-Jin;Jeon, Young-Soo;Jeong, Noh-Hee;Jeang, Bu-Shik
    • Journal of the Korean Applied Science and Technology
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    • v.27 no.3
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    • pp.233-239
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    • 2010
  • Supercritical carbondioxide is very effective in removing oils from a variety of seed matrices, devoid of any appreciable amount of phospholipid content. However, the limited solubility of phosphalipids in supercritical carbondioxide leaves behind a potentially valuable by-product in spent seed matrix. Any phospholipid extraction process from the spent matrix must maintain the structure and the quality of phospholipids and must be compatible with the end use of the seed protein meal an animal feed or for human consumption. An initial supercritical carbondioxide extraction of soybean flakes was performed at 32 MPa and $80^{\circ}C$ to extract the oils, leaving the phospholipids in the deflatted soybean flakes, A second step was performed on the defatted soybean flake using $X_{eth}$=0.10, Varying the pressure from 175 MPa to 70.6 MPa and temperature from $60^{\circ}C$ to $80^{\circ}C$. For all supercritical carbon dioxide/ethanol mixture extractions, a fraction rich in phospholipids was obtained. The fractions extracted from defatted soybean flakes were dried and them redissolved in chloroform before HPLC-ELSD analysis. Quantitative and qualitative analysis of phospholipids on soybean seeds, defatted soybean flake, and different extracted phospholipid fractions was carried out, to ascertain the effect of extraction pressure and temperature.

Angiotensin-I Converting Enzyme Inhibitory Activity of Enzymatic Hydrolysates of Food Proteins (식품단백질 효소가수분해물의 Angiotensin-I 전환효소 저해작용)

  • 염동민;노승배;이태기;김선봉;박영호
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.22 no.2
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    • pp.226-233
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    • 1993
  • Enzymatic hydrolysates of food proteins (defatted soybean cake, egg albumin and casein) were tested for inhibitory activity against angiotensin-I converting enzyme (ACE). Food proteins were hydrolysed with complex enzyme, bromelain, alcalase, $\alpha$-chymotrypsin, trypsin, papain and pepsin by heating method. The hydrolysates obtained from the treatment of complex enzyme and bromelain showed the higher ACE inhibitory activity. ACE inhibitory activity of hydrolysates exhibited a tendency to be increased until 8hrs and increased with increment of concentration. The activity was also stable by heat treatment at 10$0^{\circ}C$ for 20min. Molecular weight of active fraction was about 1, 400 and defatted soybean cake hydrolysate below 1, 400 in case of defatted soybean cake hydrolysate treated with alcalase. Amino acid of the active fractions was abundant in Asp, Glu, Lys, lle, Leu, Ala and Val.

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Antitumor Activities of Red Ginseng Acidic Polysaccharide(RGAP) as an Immunomodulator

  • Park Jong Dae;Kim Young Sook;Shin Han Jae;Park Kyung Mee;Kwak Yi Sung;Toida Toshihiko
    • Proceedings of the Ginseng society Conference
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    • 2002.10a
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    • pp.266-276
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    • 2002
  • A red ginseng acidic polysaccharide(RGAP) with immunomodulating antitumor activities was isolated from Korean red ginseng, The molecular weight of RGAP was estimated to be 12-450 kDa by gel filtration chromatography, RGAP was found to increase survival rate and to inhibit of tumor growth significantly in a dose dependent manner in mice transplanted with tumor cells. RGAP significantly promoted nitric oxide(NO) production from peritoneal macrophages bothin vivo and in vitro. Western blot analysis exhibited a newly synthesized inducible nitric oxide synthase(iNOS) protein band in the RGAP treated group. It seems likely that immunomodulating antitumor activities of RGAP are mainly mediated by NO production of macrophage. RGAP was further purified by ultrafiltration and anion exchange chromatography on DEAE-sepharose, followed by gel filtration on Sephacryl S-300 to give an active fraction(GFP) with stronger NO production in murine macrophages. GFP increased survival rate ten times compared to RGAP in male ICR mice transplanted with sarcoma 180 and also showed more potent tumoricidal activities of natural killer cells than RGAP. Sugar $composition(mol\%)$ of GFP was found to be arabinose:rhamnose:xylose:galacturonic acid:mannose:galactose:glucose(10:9:1:25:8:20:27) by GC/MS. The results suggest that clinical trials of RGAP in immunotherapy against cancer are highly feasible.

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Amelioration of experimental autoimmune encephalomyelitis by Ishige okamurae

  • Ahn, Meejung;Kim, Jeongtae;Yang, Wonjun;Choi, Yuna;Ekanayake, Poornima;Ko, Hyunju;Jee, Youngheun;Shin, Taekyun
    • Anatomy and Cell Biology
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    • v.51 no.4
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    • pp.292-298
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    • 2018
  • Experimental autoimmune encephalomyelitis (EAE) is a T-cell-mediated autoimmune central nervous system disease characterized by inflammation with oxidative stress. The aim of this study was to evaluate an anti-inflammatory effect of Ishige okamurae on EAE-induced paralysis in rats. An ethanolic extract of I. okamurae significantly delayed the first onset and reduced the duration and severity of hind-limb paralysis. The neuropathological and immunohistochemical findings in the spinal cord were in agreement with these clinical results. T-cell proliferation assay revealed that the ethyl-acetate fraction of I. okamurae suppressed the proliferation of myelin basic protein reactive T cells from EAE affected rats. Flow cytometric analysis showed $TCR{\alpha}{\beta}^+$ T cells was significantly reduced in the spleen of EAE rats with I. okamurae treatment with concurrent decrease of inflammatory mediators including tumor necrosis $factor-{\alpha}$ and cyclooxygenase-2. Collectively, it is postulated that I. okamurae ameliorates EAE paralysis with suppression of T-cell proliferation as well as decrease of pro-inflammatory mediators as far as rat EAE is concerned.

Characterization of Alpha Amylase Producing Thielaviopsis ethacetica and Its Raw Starch Hydrolyzing Ability on Different Agricultural Substrates

  • Dissanayaka, Dissanayaka M.S.;De Silva, Sembukuttige N.T.;Attanayaka, D.P.S.T.G.;Kurera, Mihidukulasuriya J.M.S.;Fernando, Charakrawarthige A.N.
    • Microbiology and Biotechnology Letters
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    • v.47 no.3
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    • pp.412-422
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    • 2019
  • The present study reports the morphological and molecular characterization of the fungal strain, CMSS06 and evaluates its raw starch hydrolyzing ability in four different agricultural substrates (rice bran, banana peel, cassava tubers, and coconut water). The potential use of each agricultural substrate to replace the expensive fermentation media was evaluated with six different fermentation media: rice bran (RB), banana peel (BP), cassava starch (CS), cassava in coconut water (CSCW), cassava in modified coconut water (CMCW), and pure Coconut water (CW). The fungal strain CMSS06 was identified as Thielaviopsis ethacetica by the analysis of the ITS sequences. The T. ethacetica alpha amylase enzyme exhibited maximum alpha amylase activity at 72 h, pH 7.0, and $40^{\circ}C$ on soluble starch. This species resulted in the highest enzyme activity (mU/ml) of 26.06, 10.89, 58.82, 14.2, and 54.67 with the RB, BP, CS, CSCW, and CMCW fermentation media, respectively. The results indicate that CS can be used as a carbon substrate and CMCW can be used to accelerate the fermentation by T. ethacetica. The enzyme was partially purified by 40-60% ammonium sulphate fraction, and it showed total enzyme activity, total protein content, specific activity, purification fold, and a recovery of 2400 mU, 30 mg, 80 mU/mg, 2.7, and 71.1%, respectively. The molecular mass of the T. ethacetica alpha amylase was estimated on SDS-PAGE, and two bands around 50 kDa and 70 kDa were identified. The present study implies that T. ethacetica can produce alpha amylase, and it can be used to hydrolyze raw starch during the fermentation processes.

Fermentation enhances the antioxidant and anti-inflammatory effects of Bat Faeces (Ye Ming Sha) via the ERK, p38 MAPK and NF-κB signaling pathways in RAW 264.7 cells

  • Lee, Han-Saem;Chon, So-Hyun;Kim, Min-A;Park, Jeong-Eun;Lim, Yu-Mi;Kim, Eun-Jeong;Son, Eun-Kyung;Kim, Sang-Jun;So, Jai-Hyun
    • Journal of Applied Biological Chemistry
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    • v.62 no.1
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    • pp.57-66
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    • 2019
  • The ethyl acetate fraction of Bat Faeces (Ye Ming Sha: natural products used in Chinese Medicine) after fermentation (EFBF-AF) showed enhanced anti-oxidative effects in 2,2-diphenyl-1-picrylhydrazyl and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt assays. Fermentation of the Bat Faeces by using the crude enzyme extract from Aspergillus kawachii, significantly increased the anti-inflammatory effects. Fermented Bat Faeces markedly inhibited nitric oxide production, inducible nitric oxide synthase, and cyclooxygenase-2 expression in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophage cells. The EFBF-AF reduced the nuclear translocation of nuclear factor kappa B ($NF-{\kappa}B$) via $IKK{\alpha}$ and $I{\kappa}B{\alpha}$ phosphorylation, and decreased the phosphorylated the extracellular signal-regulated kinases (ERK) and p38 expression in LPS-treated RAW 264.7 macrophages. In addition, the EFBF-AF suppressed the expression of pro-inflammatory genes, such as interleukin-$1{\beta}$, interleukin-6, and tumor necrosis $factor-{\alpha}$. These results suggest that fermented Bat Faeces may suppress pro-inflammatory responses in LPS-stimulated RAW 264.7 macrophages cells via ERK, p38 mitogen-activated protein kinase and $NF-{\kappa}B$ signaling pathways.

Anti-Inflammatory Effects of Grasshopper Ketone from Sargassum fulvellum Ethanol Extract on Lipopolysaccharide-Induced Inflammatory Responses in RAW 264.7 Cells

  • Kim, Min-Ji;Jeong, So-Mi;Kang, Bo-Kyeong;Kim, Koth-Bong-Woo-Ri;Ahn, Dong-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.29 no.5
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    • pp.820-826
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    • 2019
  • This study evaluated the anti-inflammatory potential of a grasshopper ketone (GK) isolated from the brown alga Sargassum fulvellum on lipopolysaccharide (LPS)-induced RAW 264.7 murine macrophage cell line. GK was isolated and purified from the n-hexane fraction and its structure was verified on the basis of NMR spectroscopic data. GK up to $100{\mu}g/ml$ is not cytotoxic to RAW 264.7, and is an effective inhibitor of LPS-induced NO production in RAW 264.7 cells. The production of pro-inflammatory cytokines, including IL-6, $IL-1{\beta}$, and $TNF-{\alpha}$ was found significantly reduced in $0.1-100{\mu}g/ml$ dose ranges of GK treatment (p < 0.05). We confirmed the dose-dependent and significant inhibition of iNOS and COX-2 proteins expression. In addition, it has been shown that GK induces anti-inflammatory effects by inhibiting MAPKs (ERK, JNK, and p38) and $NF-{\kappa}B$ p65 phosphorylation. Our results show that the anti-inflammatory properties of GK may be due to the inhibition of the $NF-{\kappa}B$ and MAPKs pathways, which are associated with the attenuation of cytokine secretion.

CircCOL1A2 Sponges MiR-1286 to Promote Cell Invasion and Migration of Gastric Cancer by Elevating Expression of USP10 to Downregulate RFC2 Ubiquitination Level

  • Li, Hang;Chai, Lixin;Ding, Zujun;He, Huabo
    • Journal of Microbiology and Biotechnology
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    • v.32 no.7
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    • pp.938-948
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    • 2022
  • Gastric cancers (GC) are generally malignant tumors, occurring with high incidence and threatening public health around the world. Circular RNAs (circRNAs) play crucial roles in modulating various cancers, including GC. However, the functions of circRNAs and their regulatory mechanism in colorectal cancer (CRC) remain largely unknown. This study focuses on both the role of circCOL1A2 in CRC progression as well as its downstream molecular mechanism. Quantitative polymerase chain reaction (qPCR) and western blot were adopted for gene expression analysis. Functional experiments were performed to study the biological functions. Fluorescence in situ hybridization (FISH) and subcellular fraction assays were employed to detect the subcellular distribution. Luciferase reporter, RNA-binding protein immunoprecipitation (RIP), co-immunoprecipitation (Co-IP), RNA pull-down, and immunofluorescence (IF) and immunoprecipitation (IP) assays were used to explore the underlying mechanisms. Our results found circCOL1A2 to be not only upregulated in GC cells, but that it also propels the migration and invasion of GC cells. CircCOL1A2 functions as a competing endogenous RNA (ceRNA) by sequestering microRNA-1286 (miR-1286) to modulate ubiquitin-specific peptidase 10 (USP10), which in turn spurs the migration and invasion of GC cells by regulating RFC2. In sum, CircCOL1A2 sponges miR-1286 to promote cell invasion and migration of GC by elevating the expression of USP10 to downregulate the level of RFC2 ubiquitination. Our study offers a potential novel target for the early diagnosis and treatment of GC.

The protective effects of Aster yomena (Kitam.) Honda on high-fat diet-induced obese C57BL/6J mice

  • Kim, Min Jeong;Kim, Ji Hyun;Lee, Sanghyun;Kim, Bohkyung;Kim, Hyun Young
    • Nutrition Research and Practice
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    • v.16 no.1
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    • pp.46-59
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    • 2022
  • BACKGROUND/OBJECTIVES: Aster yomena (Kitam.) Honda (AY) has remarkable bioactivities, such as antioxidant, anti-inflammation, and anti-cancer activities. On the other hand, the effects of AY against obesity-induced insulin resistance have not been reported. Therefore, this study examined the potential of AY against obesity-associated insulin resistance in high-fat diet (HFD)-fed mice. MATERIALS/METHODS: An obesity model was established by feeding C57BL/6J mice a 60% HFD for 16 weeks. The C57BL6/When ethyl acetate fraction from AY (EFAY) at doses of 100 and 200 mg/kg/day was administered orally to mice fed a HFD for the last 4 weeks. Normal and control groups were administered water orally. The body weight and fasting blood glucose were measured every week. Dietary intake was measured every other day. After dissection, blood and tissues were collected from the mice. RESULTS: The administration of EFAY reduced body and organ weights significantly compared to HFD-fed control mice. The EFAY-administered groups also improved the serum lipid profile by decreasing the triglyceride, total cholesterol, and low-density lipoprotein compared to the control group. In addition, EFAY ameliorated the insulin resistance-related metabolic dysfunctions, including the fasting blood glucose and serum insulin level, compared to the HFD-fed control mice. The EFAY inhibited lipid synthesis and insulin resistance by down-regulation of hepatic fatty acid synthase and up-regulation of the AMP-activated protein kinase pathway. EFAY also reduced lipid peroxidation in the liver, indicating that EFAY protected hepatic injury induced by obesity. CONCLUSIONS: These results suggest that EFAY improved obesity-associated insulin resistance by regulating the lipid and glucose metabolism, suggesting that AY could be used as a functional food to prevent obesity and insulin resistance.

Delta neutrophil index as a predictor of vesicoureteral reflux in children with febrile urinary tract infection

  • Kim, Jae Eun;Oh, Jun Suk;Yoon, Jung Min;Ko, Kyung Ok;Cheon, Eun Jung
    • Childhood Kidney Diseases
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    • v.26 no.1
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    • pp.46-51
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    • 2022
  • Purpose: Delta neutrophil index (DNI) indicates immature granulocytes in peripheral blood and has been confirmed to be effective as a prognostic factor for neonatal sepsis. Also, it has been reported to have diagnostic value in acute pyelonephritis and in predicting vesicoureteral reflux (VUR) in the infant. We conducted the study to verify whether DNI is also helpful in the entire pediatric age group with febrile urinary tract infection (UTI). Methods: Medical records of children hospitalized for febrile UTIs were analyzed retrospectively. All subjects underwent kidney ultrasound and voiding cystourethrography. In the group with and without VUR, we compared sex and age, and the following laboratory values: the white blood cell count, neutrophil, polymorphonuclear leucocyte, eosinophil, hemoglobin, platelet count, C-reactive protein, DNI value, and the finding of ultrasound. Results: A total of 315 patients (163 males and 152 females; range, 0-127 months) were eligible, and 41 patients (13%) had VUR. As a result of univariate analysis, the white blood cell count, neutrophil, DNI, and ultrasonic abnormalities were high in the reflux group, and the hemoglobin and lymphocyte fraction values were low. The value of DNI and the abnormal ultrasound were significantly higher in the reflux group on the multivariate analysis. The area under the curve value of the receiver operating curve was higher in DNI (0.640; 95% confidence interval, 0.536-0.744; P=0.004), and the DNI cutoff value for VUR prediction was 1.85%. Conclusions: We identified that ultrasound findings and DNI values were helpful predictors of VUR in pediatric febrile UTIs.