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A Novel Tannase from the Xerophilic Fungus Aspergillus niger GH1

  • Marco, Mata-Gomez;Rodriguez, Luis V.;Ramos, Erika L.;Renovato, Jacqueline;Cruz-Hernandez, Mario A.;Rodriguez, Raul;Contreras, Juan;Aguilar, Cristobal N.
    • Journal of Microbiology and Biotechnology
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    • v.19 no.9
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    • pp.987-996
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    • 2009
  • Aspergillus niger GH1 previously isolated and identified by our group as a wild tannase producer was grown under solid-state (SSC) and submerged culture (SmC) conditions to select the enzyme production system. For tannase purification, extracellular tannase was produced under SSC using polyurethane foam as the inert support. Tannase was purified to apparent homogeneity by ultrafiltration, anion-exchange chromatography, and gel filtration that led to a purified enzyme with a specific activity of 238.14 IU/mg protein with a final yield of 0.3% and a purification fold of 46. Three bands were found on the SDS-PAG with molecular masses of 50, 75, and 100 kDa. PI of 3.5 and 7.1% N-glycosylation were noted. Temperature and pH optima were 600e and 6.0 [methyl 3,4,5-trihydroxybenzoate (MTB) as substrate], respectively. Tannase was found with a $K_M$ value of $0.41{\times}10^{-4}M$ and the value of $V_{max}$ was $11.03{\mu}$moL/min at $60^{\circ}C$ for MTB. Effects of several metal salts, solvents, surfactants, and typical enzyme inhibitors on tannase activity were evaluated to establish the novelty of the enzyme. Finally, the tannase from A. niger GH1 was significantly inhibited by PMSF (phenylmethylsulfonyl fluoride), and therefore, it is possible to consider the presence of a serine or cysteine residue in the catalytic site.

Exposure and Toxicity Assessment of Ultrafine Particles from Nearby Traffic in Urban Air in Seoul, Korea

  • Yang, Ji-Yeon;Kim, Jin-Yong;Jang, Ji-Young;Lee, Gun-Woo;Kim, Soo-Hwan;Shin, Dong-Chun;Lim, Young-Wook
    • Environmental Analysis Health and Toxicology
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    • v.28
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    • pp.7.1-7.9
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    • 2013
  • Objectives We investigated the particle mass size distribution and chemical properties of air pollution particulate matter (PM) in the urban area and its capacity to induce cytotoxicity in human bronchial epithelial (BEAS-2B) cells. Methods To characterize the mass size distributions and chemical concentrations associated with urban PM, PM samples were collected by a 10-stage Micro-Orifice Uniform Deposit Impactor close to nearby traffic in an urban area from December 2007 to December 2009. PM samples for in vitro cytotoxicity testing were collected by a mini-volume air sampler with $PM_{10}$ and $PM_{2.5}$ inlets. Results The PM size distributions were bi-modal, peaking at 0.18 to 0.32 and 1.8 to $3.2{\mu}m$. The mass concentrations of the metals in fine particles (0.1 to $1.8{\mu}m$) accounted for 45.6 to 80.4% of the mass concentrations of metals in $PM_{10}$. The mass proportions of fine particles of the pollutants related to traffic emission, lead (80.4%), cadmium (69.0%), and chromium (63.8%) were higher than those of other metals. Iron was the dominant transition metal in the particles, accounting for 64.3% of the $PM_{10}$ mass in all the samples. We observed PM concentration-dependent cytotoxic effects on BEAS-2B cells. Conclusions We found that exposure to $PM_{2.5}$ and $PM_{10}$ from a nearby traffic area induced significant increases in protein expression of inflammatory cytokines (IL-6 and IL-8). The cell death rate and release of cytokines in response to the $PM_{2.5}$ treatment were higher than those with $PM_{10}$. The combined results support the hypothesis that ultrafine particles from vehicular sources can induce inflammatory responses related to environmental respiratory injury.

A case of idiopathic pulmonary hemosiderosis with seasonal recurrence (계절성으로 재발한 특발성 폐 혈철 침착증 1예)

  • Kwak, Ga Young;Lee, Na Young;Lee, Moon Hee;Lee, Soo Young;Chung, Seung Yun;Kang, Jin Han;Jeong, Dae Chul
    • Clinical and Experimental Pediatrics
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    • v.52 no.2
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    • pp.256-260
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    • 2009
  • Idiopathic pulmonary hemosiderosis (IPH) is a rare disease affecting mostly children. This disorder is characterized by recurrent episodes of hemoptysis, bilateral diffuse pulmonary infiltrates, and iron-deficiency anemia. An acute fulminant alveolar hemorrhage can be fatal due to respiratory failure, while chronic hemorrhage leads to hemosiderin-laden macrophages and pulmonary fibrosis. Genetic, autoimmune, allergic, environmental, and metabolic mechanisms of pathogenesis have been suggested, but the etiology of IPH remains unknown. We report on a 9-year-old girl with idiopathic pulmonary hemosiderosis who showed seasonal recurrences without cause.

Effects of plant-based Korean food extracts on lipopolysaccharide-stimulated production of inflammatory mediators in vitro

  • Lee, Sun Young;Kim, Yoo-Sun;Lim, Ji Ye;Chang, Namsoo;Kang, Myung-Hee;Oh, Se-Young;Lee, He-Jin;Kim, Hyesook;Kim, Yuri
    • Nutrition Research and Practice
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    • v.8 no.3
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    • pp.249-256
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    • 2014
  • BACKGROUND/OBJECTIVES: The traditional Korean diet is plant-based and rich in antioxidants. Previous studies have investigated the potential health benefits of individual nutrients of Korean foods. However, the cumulative effects of a Korean diet on inflammation remain poorly understood. Therefore, the aim of this study was to investigate the anti-inflammatory effects of a plant-based Korean diet. MATERIALS/METHODS: Using data from the Fifth Korean National Health and Nutrition Examination Survey, 75 individual plant food items were selected which represent over 1% of the total diet intake of the Korean diet. These items were classified into ten different food groups, and the vegetable (Veg) and fruit (Fruit) groups were studied based on their high antioxidant capacity. For comparison, a mixture of all ten groups (Mix) was prepared. To produce a model of inflammation with which to test these Veg, Fruit, and Mix plant-based Korean food extracts (PKE), RAW264.7 macrophages were treated with lipopolysaccharide (LPS). RESULTS: Levels of nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$), as well as protein expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) were found to be lower following PKE treatment. Furthermore, PKE treatment was found to suppress tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) and interleukin-6 (IL-6) via the nuclear transcription factor kappa-B ($NF-{\kappa}B$) signaling pathway. Overall, the Mix group exhibited the greatest anti-inflammatory effects compared with Veg and Fruit PKE group. CONCLUSIONS: Inhibition of LPS-induced pro-inflammatory mediators by the PKE tested was found to involve an inhibition of NF-kB activation. Moreover, PKE tested have the potential to ameliorate various inflammation-related diseases by limiting the excessive production of pro-inflammatory mediators.

Comparison of Nutritional and Physical Status according to the Residental Type among College Women in Seoul Women's University and Sahmyook University (삼육대 및 서울여대 재학 중인 일부 여대생의 거주형태에 따른 영양섭취상태 및 신체계측 비교 연구)

  • Choi, Kyung-Soon;Shin, Kyung-Ok;Huh, Seon-Min;Chung, Keun-Hee
    • Journal of Nutrition and Health
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    • v.43 no.1
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    • pp.86-96
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    • 2010
  • This study was conducted to investigate the effects of residence types ((1) parent house, (2) dormitory or boarding home, (3) boarding home w/o food) on the dietary habits and health status of college women (20-23 y). Overall, 46.1% of college women reported that they were economically middle class and spent 200,000-390,000 won. Additionally, 17.4% of college women considered themselves healthy, while 33.8% reported that they were not healthy. Furthermore, 32.0% of college women had experience to control their weight and 39.8% reported that they exercised to control their weight. There were no significant differences in the height and weight of the subjects according to residential type, but the amount of skeletal muscles mass ($21.0\;{\pm}\;2.6\;kg$) of the subjects that lived in dormitory or boarding home was significantly higher (p < 0.05) than that of women who lived at home. Additionally, 30.8% of students that lived in private residences, 25.0% of students that dwelled in dormitory or boarding homes and 27.7% of students that boarding home w/o food had three regular meals every day (p < 0.05). It has been reported that 18.3% of college students eat processed and instant foods due to their convenience. In the present study, intakes of energy, protein, fat, vitamin A, vitamin $B_1$, vitamin $B_2$, vitamin $B_6$, niacin, vitamin C, vitamin E and folic acid for the subjects who lived in dormitory or boarding homes was significantly higher than those of students who lived in private residences (p < 0.05). Additionally, intakes of iron, calcium, and zinc were lower in the subjects that boarding home w/o food (p < 0.05). Finally, the blood glucose level was $84.7\;{\pm}\;13.0\;mg/dL$, and differed significantly by residential types (p < 0.05); however, the average glucose levels of all subjects were within the normal range (90-110 mg/dL).

Endotoxin-induced Acute Lung Injury is Mediated by PAF Produced via Remodelling of Lyso PAF in the Lungs

  • Lee, Young-Man;Kim, Teo-An
    • The Korean Journal of Physiology and Pharmacology
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    • v.4 no.3
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    • pp.219-226
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    • 2000
  • In order to elucidate the role of platelet activating factor (PAF) in the acute lung injury induced by endotoxin (ETX), activities of phospholipase A2, lyso PAF acetyltransferase and oxidative stress by neutrophilic respiratory burst were probed in the present study. To induce acute lung injury, $100\;{\mu}g$ of E.coli ETX (type 0127; B8) was instilled directly into the tracheae of Sprague-Dawley rats. Five hours after the ETX instillation, induction of acute lung injury was confirmed by lung leak index and protein contents in the bronchoalveolar lavage (BAL) fluid. At the same time, lung phospholipase A2 (PLA2) activity and expression of group I and II secretory type PLA2 were examined. In these acutely injured rats, ketotifen fumarate, known as lyso PAF acetyltransferase inhibitor and mepacrine were administered to examine the role of PAF in the pathogenesis of the acute lung injury. To know the effect of the ETX in the synthesis of the PAF in the lungs, lyso PAF acetyltransferase activity and PAF content in the lungs were measured after treatments of ETX, ketotifen fumarate and mepacrine. In addition, the role of neutrophils causing the oxidative stress after ETX was examined by measuring lung myeloperoxidase (MPO) and enumerating neutrophils in the BAL fluid. To confirm the oxidative stress in the lungs, pulmonary contents of malondialdehyde (MDA) were measured. After instillation of the ETX in the lungs, lung leak index increased dramatically (p<0.001), whereas mepacrine and ketotifen decreased the lung leak index significantly (p<0.001). Lung PLA2 activity also increased (p<0.001) after ETX treatment compared with control, which was reversed by mepacrine and ketotifen (p<0.001). In the examination of expression of group I and II secretory PLA2, mRNA synthesis of the group II PLA2 was enhanced by ETX treatment, whereas ketotifen and WEB 2086, the PAF receptor antagonist, decreased the expression. The activity of the lysoPAF acetyltransferase increased (p<0.001) after treatment of ETX, which implies the increased synthesis of PAF by the remodelling of lysoPAF in the lungs. Consequently, the contents of the PAF in the lungs were increased by ETX compared with control (p<0.001), while mepacrine (p<0.001) and ketotifen (p<0.01) decreased the synthesis of the PAF in the lungs of ETX treated rats. The infiltration of the neutrophils was confirmed by measuring and enumerating lung MPO and the neutrophils in the BAL fluid respectively. Compared with control, ETX increased lung MPO and number of neutrophils in BAL significantly (p<0.001) whereas mepacrine and ketotifen decrerased number of neutrophils (p<0.001) and MPO (p<0.05, p<0.001, respectively). The lung MDA contents were also increased (p<0.001) by ETX treatment, but treatment with mepacrine (p<0.001) and ketotifen (p<0.01) decreased the lung MDA contents. Collectively, we conclude that ETX increases PLA2 activity, and that the subsequently increased production of PAF was ensued by the remodelling of the lyso PAF resulting in tissue injury by means of oxidative stress in the lungs.

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Program Analysis Activities for Development of Componentization Methodology for of Legacy System (레가시 시스템의 컴포넌트화 방법론 개발을 위한 프로그램 분석 활동)

  • Cha, Jung-Eun;Kim, Cheol-Hong;Yang, Young-Jong
    • Proceedings of the Korea Information Processing Society Conference
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    • 2002.11c
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    • pp.2119-2122
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    • 2002
  • 기업의 비즈니스 프로세스가 복잡, 다양해짐에 따라, 현재 운영 시스템에 대한 급격한 기술적 변화를 수용하고 이를 조직적 측면의 기업 프로세스로 적용하기 위해 레가시 시스템의 현대화가 요구된다. 따라서, 현재의 기업들은 다양한 사용자들이 각자 그들의 관점에서 필요한 비즈니스 요구들을 웹 상에서 처리시킬 수 있도록 J2EE, .NET 등으로 대표되는 컴포넌트 및 웹 서비스 기술을 적용한 새로운 e-business 환경을 수용해야만 한다. 하지만, 기업 조직의 중요한 지식과 프로세스들을 처리하는 시스템들은 대부분 과거(Legacy)의 기술에 의해 개발되어졌으며, 이러한 시스템들을 새로운 비즈니스 환경에 적용하기에는 웹 환경을 위한 분산 아키텍쳐의 결여와 개방성과 표준화 미흡으로 시스템의 유지보수에 많은 어려움을 가진다. 또한, 방법론 차원에서 재공학의 절차와 기법을 체계적으로 정의하고 지원하기 위한 노력이 매우 부족한 실정이다. 본 논문에서는 레거시 시스템을 새로운 시스템 환경으로의 변환 및 통합을 위한 재공학 방법론 개발을 목적으로 프로그램 분석 활동을 설명한다. 본 논문에서 개발하고자 하는 방법론은 다양한 추상화 수준에서 역공학 정보를 복구하고, 컴포넌트화 단계를 통해 새로운 시스템으로 진화할 수 있는 절차 및 기법들을 제공한다. 레거시 시스템 컴포넌트화 방법론은 변환계획 단계, 역공학 단계, 컴포넌트화 단계, 인도 단계의 4 단계로 구성되어 있으며, 본 논문에서는 전체 단계 중 가장 기초가 되고 중요한 단계인 역공학 단계에 초점을 두고 프로그램 분석을 위한 절차 및 과정의 주요 지침들을 제시한다.0보다 유의적으로 우수하였으며, 맛, 다즙성 및 전체적인 기호도는 TMR-0 및 TMR-1 사이에 유의성이 없었다(p<0.05).能性)을 알아보고자 본(本) 실험(實驗)을 실시(實施)한 결과(結果), 유의성(有意性) 있는 결과(結果)를 얻었기에 보고(報告)하는 바이다.또한 이들 상피세포(上皮細胞)들을 투과전자현미경적(透過電子顯微鏡的)으로 관찰(觀察)하였을 때 초미세구조(超微細構造)가 잘 보존(保存)되었으나 Langer-hans cell내(內)의 Birbeck granule은 유리전(遊離前) 피부상피조직내(皮膚上皮組織內)의 Langerhans cell내(內)의 Birbeck granule에 비(比)해 수적(數的)으로 현저히 감소(減少)되어 있었다. 그러나 Thy-1 양성(陽性) dendritic cell에서 볼 수 있는 dense-core 과립(顆粒)은 별변화(別變化)없이 쉽게 관찰(觀察)될 수 있었다. 조직배양(組織培養)을 한 견(犬)의 keratinocyte에 대(對)해 사람 pemphigus vulgaris의 항체(抗體)로 반응(反應)시킨 후 protein-A gold(15 nm)로 표식(標識)시킨 바 제일 바깥 상층(上層)의 keratinocyte에 있어서 세포막표면(細胞膜表面)을 따라 표식(標識)되어 세포막항원(細胞膜抗元)을 나타내었으며, 이와 같은 소견(所見)으로 미루어 정상피부(正常皮膚) 중층편평상피세포(重層扁平上皮細胞)에서도 동일(同一)한 소견(所見)을 관찰(觀察)할 수 있다고 본다.al remnants, Resorption of fetus로 관찰된 것이다. Fetal death는 수정후

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Downregulation of $\textrm{NF}_{-{\kappa}}\textrm{B}$ activation in human keratinocytes by melanogenic inhibitors

  • Ahn, Kwang-Seok;Lee, Jinseon;Kim, Yeong-Shik
    • Proceedings of the SCSK Conference
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    • 2003.09a
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    • pp.780-803
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    • 2003
  • Exposure of skin cells, particularly keratinocytes to various nuclear factor-kappaB ($\textrm{NF}_{-{\kappa}}\textrm{B}$) activators [e.g. tumor necrosis factor-$\alpha$, interleukin-1, lipopolysaccharides, and ultraviolet light] leads to phosphorylation and degradation of the inhibitory protein, $\textrm{I}_{{\kappa}}\textrm{B}$. Liberated $\textrm{NF}_{-{\kappa}}\textrm{B}$ is translocated into the nucleus where it can change or alter expression of target genes, resulting in the secretion of extracellular signaling molecules including melanotrophic factors affecting melanocyte. In order to demonstrate the possible role of $\textrm{NF}_{-{\kappa}}\textrm{B}$ activation on the synthesis of melanotrophic factors from the keratinocytes, the activities of $\textrm{NF}_{-{\kappa}}\textrm{B}$ induced by melanogenic inhibitors (MIs) were determined in human HaCaT keratinocytes transfected with $\textrm{pNF}_{-{\kappa}}\textrm{B}$-SEAP-NPT plasmid. Transfectant cells released the secretory alkaline phosphatase (SEAP) as a transcription reporter in response to the $\textrm{NF}_{-{\kappa}}\textrm{B}$ activity and contain the neomycin phosphotransferase (NPT) gene for the dominant selection marker for geneticin resistance. MIs such as niacinamide, kojic acid, hydroquinone, resorcinol, arbutin, and glycolic acid were preincubated with transfectant HaCaT cells for 3 h and then ultraviolet B (UVB) was irradiated. $\textrm{NF}_{-{\kappa}}\textrm{B}$ activation was measured with the SEAP reporter gene assay using a fluorescence detection method. Of the Mis tested, kojic acid ($IC_{50}$/ = 60 $\mu$M) was found to be the most potent inhibitor of UVB-upregulating $\textrm{NF}_{-{\kappa}}\textrm{B}$ activation in transfectant HaCaT cells, which is followed by niacinamide ($IC_{50}$/= 540 $\mu$M). Pretreatment of the transfectant HaCaT cells with the Mis, especially kojic acid and niacinamide, effectively lowered $\textrm{NF}_{-{\kappa}}\textrm{B}$ binding measured by electrophoretic mobility shift assay. Furthermore, these two inhibitors remarkably reduced the secretion level of IL-6, one of melanotrophic factors, triggered by UV-radiation of the HaCaT cells. These observations suggest that Mis working at the in vivo level might act partially through the modulation of the synthesis of melanotrophic factors in keratinocyte.

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Single and 28-day repeated dose toxicity studies of botulinum toxin type A in mice and rats (마우스 및 랫드에서 botulinum toxin type A의 단회 및 28일 반복투여 독성시험)

  • Jeon, Tae-Won;Kim, Ji-Young;Hyun, Sun-Hee;Kim, Nam-Hee;Lee, Sang-Kyu;Kim, Chun-Hwa;Woo, Hee-Dong;Yang, Gi-Hyeok;Jung, Hyun-Ho;Jeong, Tae-Cheon
    • Korean Journal of Veterinary Research
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    • v.43 no.1
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    • pp.57-66
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    • 2003
  • Single and 28-day repeated dose toxicity studies of botulimnn toxin type A were carried out in ICR mice and Sprague-Dawley rats, respectively. In the single dose toxicity study, botulinwn toxin was injected intraperitoneally to male and female mice at a single dose of 40, 59, 89 133 and 200 ng/10 ml saline/kg. All animals died from 59 ng/kg group. Some clinical signs, such as decrease in locomotor activity, dyspnea, prone position and ptosis, were observed in most of both sexes from 59 ng/kg group, but no signs were seen in all animals at 40 ng/kg group. The results showed that the median lethal dose of botulinum toxin might be in the range of 40-59 ng/kg in both sexes. In the repeated dose toxicity study, the test material was administered intradermally for 28 days at doses of 0 (vehicle-treated control), 1.25, 2.5, 5.0 and $10.0ng/head/50{\mu}{\ell}$ saline in male and female rats. No test material-related changes were noted in survivals, clinical signs, food and water consumptions and gross finding in any group. Botulinum toxin treatment significantly decreased the body weight gain rate in male of 5.0 ng/head group and over and in female of 10.0 ng/head group compared to vehicle-treated control. One or more relative organ weights (i.e., spleen, thymus, liver and kidney) were increased significantly from 5.0 ng/head group compared to vehicle-treated control in both sexes. Serum biochemistry revealed increases in aspartate aminotransferase (AST), alanine aminotransferase (ALT), creatine phosphokinase, total protein and albumin in male, and increases in AST and ALT and decreases in $K^+$ and $Cl^-$ in female without dose-pendent manners. In the histopathological study, physical stimulation by needle caused slight inflammations of dennis. In addition, botulinum toxin treatment induced denervation of nerve cell and disuse of muscle, resulting in atrophy of skeletal muscle in both sexes from 2.5 ng/head group. When the antibodies to toxin were determined in all animals, a significant increase in serum antibodies was observed from 5.0 ng/head group. The results showed that the NOAEL of botulinum toxin might be 1.25 ng/head for 28-day repeated dose toxicity in rats.

Nutritional composition and functionality of mixed cereals powder (혼합잡곡분말의 영양성 및 기능성)

  • Kim, Ja-Min;Park, Joo-Young;Kim, Kun-Woo;Yoon, Kyung-Young
    • Food Science and Preservation
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    • v.21 no.3
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    • pp.388-395
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    • 2014
  • Six types of mixed cereals powder were prepared using different mixing ratios of barley, proso millet, sorghum, and adlay, and their nutritional components and functionalities were analyzed. The mixed cereals powders contained high levels of crude protein. The free amino acid and mineral contents of Sample E were the highest among all the samples. Fructose, glucose, sucrose, and maltose were detected as the four major free sugars in all the mixed cereals powders, and Sample D had the highest amount of reducing sugar among all the samples. The total polyphenol and total flavonoid contents of Sample F were significantly higher than those of the other samples. Sample C showed the highest level of DPPH radical scavenging activity and reducing power. As for the ABTS radical scavenging activity, the $IC_{50}$ of Sample D was the lowest among all the samples, but did not significantly differ from that of Sample C. Therefore, we suggest 2:2:1:1 (in Sample D) as the best mixing ratio of barley, proso millet, sorghum, and adlay to produce a mixed cereals powder with excellent nutritional value and antioxidant activities.