• Title/Summary/Keyword: Chemical assay

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The research on antioxidative effect of Sasa quelpaertensis extractum and assessment of cytotoxicity (제주조릿대 잎 추출물의 항산화효능 및 세포독성 평가에 관한 연구)

  • Lee, Song Hee;Lee, Moo Sung
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.18 no.3
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    • pp.687-693
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    • 2017
  • This research analyzed cell survival rate according to antioxidative effect and cytotoxicity using Sasa quelpaertensis and Ethanol extracts. Sasa quelpaertensis used extracts of $95^{\circ}C$ boiled-water extract and 70% ethanol liquid each. For total polyphenol content, tannin content, and flavonoid content, polyphenol content of $95^{\circ}C$ boiled-water extract was 26.6 mg/g while that of 70% ethanol extract liquid was 22.3 mg/g, which means polyphenol content was higher in $95^{\circ}C$ boiled-water extract. Tannin content was 72.1 mg/g in $95^{\circ}C$ boiled-water extract which was higher than 61.2 mg/g in 70% ethanol extract liquid. Total flavonoid content was higher in 70% ethanol extract liquid (25.4 mg/g) than in $95^{\circ}C$ boiled-water extract (17.8 mg/g). Further, the researcher measured DPPH radical elimination and ABTS radical elimination. As a result, as the concentration of each extract increased, DPPH radical elimination and ABTS radical elimination increased. In the DPPH radical elimination, L-ascorbic acid was eliminated at 500 ppm and showed no change even though the concentration increased, whereas it increased in BHT and Sasa quelpaertensis leaf extracts according to concentration. ABTS radical elimination indicated the similar phenomenon, whereas BHT showed maximum elimination at 500 ppm and decreased gradually as the concentration increased. On the other hand, it gradually increased in Sasa quelpaertensis leaf extract and showed 90% elimination at 10,000 pm, and it increased in $95^{\circ}C$ boiled-water extract. Using human skin fibroblasts (3T3) to check the cell survival rate of Sasa quelpaertensis leaves showed that cell survival rate decreased between 85.6 and 66.6% in $95^{\circ}C$ boiled-water extract. The cell survival rate was between 95.4 and 92.1% when using 70% ethanol extracts. Sasa quelpaertensis has efficacy as a natural antioxidant as well as a material hyangjang.

The Effect of Histamine on Polymorphonuclear Leukocyte-induced Pneumocyte Injury in Vitro (다형핵구에 의한 폐포세포 손상에 Histamine이 미치는 영향)

  • Kim, Young-Kyoon;Kwon, Soon-Seog;Kim, Kwan-Hyung;Han, Ki-Don;Moon, Hwa-Sik;Sang, Jeong-Sup;Park, Sung-Hak
    • Tuberculosis and Respiratory Diseases
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    • v.39 no.3
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    • pp.228-235
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    • 1992
  • Background: Although polymorphonuclear leukocytes (PMN) are important in protecting the airways and alveolar surfaces, there is evidence that they can also injure the lung while exercising their defensive role. However it has been unclear whether PMN-induced pneumocyte injury is mediated by their direct cytotoxic effect on target cells or by PMN-derived cytotoxic mediators. On the other hand histamine was known not only to act as an important chemical mediator participated in the pathogenesis of some atotic and allegic disorders, but also to have an inhibitory effect on normal PMN functions. Method: To study the mechanism by which PMN induce pneumocyte injury, we cocultured PMN from four healthy nonsmokers or their PMN-derived supernatants (PMN-SPN) with monolayers of $^{51}Cr$-labeled human A549 pneumocytes and compared PMN-and PMN-SPN-mediated pneumocyte injuries measured by $^{51}Cr$ release assay. We also compared the effects of histamine on each pneumocyte injury. Results: 1) PMN-SPN showed more injurious effect on A549 pneumocytes than that of PMN itself regardless histamine pretreatment of PMN. 2) Pneumocyte injury by PMN with histamine pretreatment was increased or decreased compared with that by PMN without histamine pretreatment, according to histamine concentrations, and PMN stimulating agents and their concentrations. 3) Pneumocyte injury by PMN-SPN with histamine pretreatment tended to be decreased compared with that by PMN-SPN without histamine pretreatment. Conclusion: Our results suggest that PMN-SPN may play more important role in mediating pneumocyte injury than PMN itself and that histamine may partially play a protective role on PMN-induced pneumocyte injury. Alternatively we conclude that the effects of histamine on PMN-induced pneumocyte injury may be affected by microenvironment in vivo.

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Quality characteristics of ground coffee extracts according to physical and chemical defects (커피빈의 물리화학적 결점에 따른 추출액의 품질특성)

  • Sung, Jun-Hyung;Cho, Jeong-Seok;Lee, Hyeon-Jeong;Choi, Ji-Young;Lee, Yeong-Min;Luo, Jin;Moon, Kwang-Deog
    • Food Science and Preservation
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    • v.23 no.5
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    • pp.638-644
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    • 2016
  • To examine the utilization possibility of defective coffee beans, non-defective and defective coffee bean were compared by means of its physiochemical properties and antioxidant capacities measured by DPPH radical scavenging activity, FRAP assay, total phenol contents, functional component (trigonelline, caffeine, chlorogenic acid) contents. After roasting process, pH and soluble solid contents of coffee extracts decreased; $L^*$ value decreased while $a^*$ and $b^*$ values increased. DPPH radical scavenging activities of defective green bean extracts were higher than that of non-defective green bean extracts. Immature green bean extract showed the highest radical scavenging activity. In FRAP assay, green bean extracts ranged from 15.28~21.80 mM TE which was higher than roasted bean extracts which showed 14.81~16.38 mM TE. Total phenol contents of green bean extracts ranged 191.06~256.25 mg% GAE which was higher than that of roasted bean extracts showed 161.91~173.44 mg% GAE. The contents of trigonelline, caffeine, chlorogenic acid in immature green bean extract were the highest, which showed 895.20 mg/L, 825.85 mg/L and 3,836.94 mg/L respectively. Each contents were decreased after roasting process. Results of this study suggest that defective coffee bean can be used as a functional food material.

Changes in phytoestrogen contents and antioxidant activities during fermentation of soybean-powder milks prepared from different soybean cultivars by Lactobacillus plantarum P1201 (Lactobacillus plantarum P1201에 의한 콩 품종별 콩-분말 두유 발효 과정에서의 식물성 에스트로젠 함량과 항산화 활성의 변화)

  • Hwang, Chung Eun;Haque, Md. Azizul;Lee, Jin Hwan;Ahn, Min Ju;Lee, Hee Yul;Lee, Byong Won;Lee, Yu-Young;Lee, Choonwo;Kim, Byung Joo;Park, Ji-Yong;Sim, Eun-Yeong;Lee, Dong Hoon;Ko, Jong Min;Kim, Hyun Tae;Cho, Kye Man
    • Korean Journal of Microbiology
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    • v.52 no.2
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    • pp.202-211
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    • 2016
  • This study evaluated the changes of phytoestrogen contents and antioxidant activities of soybean-powder milk (SPM) prepared from yellow soybean during fermentation with Lactobacillus plantarum P1201. In consequence, the levels of total phenolic and isoflavone-aglycone contents, ABTS and DPPH radical-scavenging activities, and FRAP assay values increased, while isoflavone-glycoside contents decreased during fermentation. The highest levels of daidzein, glycitein, and genistein were present in the Daepung SPM at concentrations of 177.92, 20.64, and $106.14{\mu}g/g$, respectively after 60 h of fermentation. Moreover, Daepung SPM showed the highest DPPH radical-scavenging activity of 48.54%, an ABTS radical-scavenging activity of 99.25%, and a FRAP assay value of 0.84 at the end of fermentation. The fermented Daepung SPM possessed highest isoflavone aglycone contents and antioxidant activities, which can be utilized for the development of functional foods.

Activation of Antioxidant-Response Element (ARE), Mitogen- Activated Protein Kinases (MAPKs) and Caspases by Major Green Tea Polyphenol Components during Cell Survival and Death

  • Chen, Chi;Yu, Rong;Owuor, Edward D.;Kong, A.NTony
    • Archives of Pharmacal Research
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    • v.23 no.6
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    • pp.605-612
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    • 2000
  • Green tea polyphenols (GTP) have been demonstrated to suppress tumorigenesis in several chemical-induced animal carcinogenesis models, and predicted as promising chemopreventive agents in human. Recent studies of GTP extracts showed the involvement of mitogen-activated protein kinases (MAPKs) in the regulation of Phase II enzymes gene expression and induction of apoptosis. In the current work we compared the biological actions of five green tea catechins: (1) induction of ARE reporter gene, (2) activation of MAP kinases, (3) cytotoxicity in human hepatoma HepG2-C8 cells, and (4) caspase activation in human cervical squamous carcinoma HeLa cells. For the induction of phase IIgene assay, (-)-epigallocatechin-3-gallate (EGCG) and (-)-epicatechin-3-gallate (ECG) potently induced antioxidant response element (ARE)-mediated luciferase activity, with induction observed at 25 $\mu\textrm{m}$with EGCG. The induction of ARE reporter gene appears to be structurally related to the 3-gallate group. Comparing the activation of MAPK by the five polyphenols, only EGCG showed potent activation of all three MAPKs (ERK, JNK and p38) in a dose- and time-dependent manner, whereas EGC activated ERK and p38. In the concentration range of 25 $\mu\textrm{m}$ to 1 mM, EGCG and ECG strongly suppressed HepG2-ARE-C8 cell-growth. To elucidate the mechanisms of green tea polyphenol-induced apoptosis, we measured the activation of an important cell death protein, caspase-3 induced by EGCG, and found that caspase-3 was activated in a dose- and time-dependent manner. Interestingly, the activation of caspase-3 was a relatively late event (peaked at 16 h), whereas activation of MAPKs was much earlier (peaked at 2 h). It is possible, that at low concentrations of EGCG, activation of MAPK leads to ARE-mediated gene expression including phase II detoxifying enzymes. Whereas at higher concentrations of EGCG, sustained activation of MAPKs such as JNK leads to apoptosis. These mechanisms are currently under investigation in our laboratory. As the most abundant catechin in GTP extract, we found that EGCG potently induced ARE-mediated gene expression, activated MAP kinase pathway, stimulated caspase-3 activity, and induced apoptosis. These mechanisms together with others, may contribute to the overall chemopreventive function of EGCG itself as well as the GTP.

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Evaluation of Amino Acid and Energy Utilization in Feedstuff for Swine and Poultry Diets

  • Kong, C.;Adeola, O.
    • Asian-Australasian Journal of Animal Sciences
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    • v.27 no.7
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    • pp.917-925
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    • 2014
  • An accurate feed formulation is essential for optimizing feed efficiency and minimizing feed cost for swine and poultry production. Because energy and amino acid (AA) account for the major cost of swine and poultry diets, a precise determination of the availability of energy and AA in feedstuffs is essential for accurate diet formulations. Therefore, the methodology for determining the availability of energy and AA should be carefully selected. The total collection and index methods are 2 major procedures for estimating the availability of energy and AA in feedstuffs for swine and poultry diets. The total collection method is based on the laborious production of quantitative records of feed intake and output, whereas the index method can avoid the laborious work, but greatly relies on accurate chemical analysis of index compound. The direct method, in which the test feedstuff in a diet is the sole source of the component of interest, is widely used to determine the digestibility of nutritional components in feedstuffs. In some cases, however, it may be necessary to formulate a basal diet and a test diet in which a portion of the basal diet is replaced by the feed ingredient to be tested because of poor palatability and low level of the interested component in the test ingredients. For the digestibility of AA, due to the confounding effect on AA composition of protein in feces by microorganisms in the hind gut, ileal digestibility rather than fecal digestibility has been preferred as the reliable method for estimating AA digestibility. Depending on the contribution of ileal endogenous AA losses in the ileal digestibility calculation, ileal digestibility estimates can be expressed as apparent, standardized, and true ileal digestibility, and are usually determined using the ileal cannulation method for pigs and the slaughter method for poultry. Among these digestibility estimates, the standardized ileal AA digestibility that corrects apparent ileal digestibility for basal endogenous AA losses, provides appropriate information for the formulation of swine and poultry diets. The total quantity of energy in feedstuffs can be partitioned into different components including gross energy (GE), digestible energy (DE), metabolizable energy (ME), and net energy based on the consideration of sequential energy losses during digestion and metabolism from GE in feeds. For swine, the total collection method is suggested for determining DE and ME in feedstuffs whereas for poultry the classical ME assay and the precision-fed method are applicable. Further investigation for the utilization of ME may be conducted by measuring either heat production or energy retention using indirect calorimetry or comparative slaughter method, respectively. This review provides information on the methodology used to determine accurate estimates of AA and energy availability for formulating swine and poultry diets.

Antioxidative and Hepatoprotective Effects of Injinho-Tang and Osumogwa-Tang (인진호탕(茵蔯蒿湯)과 오수목과탕(吳茱木瓜湯)의 항산화(抗酸化) 및 간세포(肝細胞) 보호효과(保護效果))

  • Lee, Sang-Hyun;Kim, Young-Bok
    • Herbal Formula Science
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    • v.16 no.1
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    • pp.117-130
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    • 2008
  • Korean traditional medicine has been used for the treatment of the various diseases based on both oriental medicinal theory and clinical trials. Thus, the prescriptions of Korean traditional medicine would be useful for the development of new therapeutics. This research focuses on the fundamental study in Korean traditional prescriptions for the development of new hepatoprotective agents. We found two prescriptions. Injinho-Tang and Osumogwa-Tang, showed the significant DPPH free radical scavenging and hepatoprotective effect, respectively. It is well-known that free radical scavenging effect is related to the prevention of various pathological events including liver injury. This paper deals with hepatoprotective effects on tacrine-induced cytotoxicity in Hep G2 cells, free radicals scavenging on both DPPH and superoxide of above two prescriptions. Hot water extract of Injinho-Tang did not show the significant hepatoprotective effect on tacrine-induced cytotoxicity in Hep G2 cells, however, it shows the significant scavenging effects for both DPPH and superoxide radicals. On the other hand, all of the hot water extracts of constituent herbal drugs in Injinho-Tang exhibited the promising protective effect on tacrine-induced cytotoxicity in Hep G2 cells. Of these, water extract of Rhei Rhizoma showed the most prominent effect on tacrine-induced cytotoxicity in Hep G2 cells. Bioassay-guided fractionation of Rhei Rhizoma extract has furnished four compounds, and their chemical structures have been identified by comparison of their spectral data with those of literature as chrysophanol (1), emodin (2), 3,5-dihydroxy-4'- methoxystilbene (3), and rhapontigenin (4), respectively. Among the isolated compounds, compounds 2-4 revealed the significant hepatoprotective effect in vitro when their $EC_{50}$ values compare with that of silybin, as a positive control. It also exhibited that emodin possessed the most hepatoprotective effect among these active compounds. In case of Osumogwa-Tang, its hot water extract showed the moderate protective effect on tacrine-induced cytotoxicity in Hep G2 cells. Hot water extract of Chaenomelis Fructus, one of the constituent herbal drug of this prescription, exhibited the significant hepatoprotective effect with $EC_{50}$ value of $7.8{\pm}0.1\;{\mu}g/ml$, however, it showed strong cytotoxicity in Hep G2 cells above the concentration of $25\;{\mu}g/ml$. It was revealed that both hot water extract of Evodiae Fructus and its butanol soluble fraction showed the moderate hepatoprotective effect but concentration-dependent activity in Hep G2 assay system. Two quinolone alkaloids, evocarpine and dihydroevocarpine, also tested for their hepatoprotective effects on tacrine-induced cytotoxicity in Hep G2 cells, however, these two compounds derived from the Evodiae Fructus as the major constituents did not show in vitro hepatoprotective effect. From these results, it would be necessary to further isolation of its hepatoprotective compounds from the butanol soluble fraction of the hot water extract of Evodiae Fructus.

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Surface characteristics of thermally treated titanium surfaces

  • Lee, Yang-Jin;Cui, De-Zhe;Jeon, Ha-Ra;Chung, Hyun-Ju;Park, Yeong-Joon;Kim, Ok-Su;Kim, Young-Joon
    • Journal of Periodontal and Implant Science
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    • v.42 no.3
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    • pp.81-87
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    • 2012
  • Purpose: The characteristics of oxidized titanium (Ti) surfaces varied according to treatment conditions such as duration time and temperature. Thermal oxidation can change Ti surface characteristics, which affect many cellular responses such as cell adhesion, proliferation, and differentiation. Thus, this study was conducted to evaluate the surface characteristics and cell response of thermally treated Ti surfaces. Methods: The samples were divided into 4 groups. Control: machined smooth titanium (Ti-S) was untreated. Group I: Ti-S was treated in a furnace at $300^{\circ}C$ for 30 minutes. Group II: Ti-S was treated at $500^{\circ}C$ for 30 minutes. Group III: Ti-S was treated at $750^{\circ}C$ for 30 minutes. A scanning electron microscope, atomic force microscope, and X-ray diffraction were used to assess surface characteristics and chemical composition. The water contact angle and surface energy were measured to assess physical properties. Results: The titanium dioxide ($TiO_2$) thickness increased as the treatment temperature increased. Additional peaks belonging to rutile $TiO_2$ were only found in group III. The contact angle in group III was significantly lower than any of the other groups. The surface energy significantly increased as the treatment temperature increased, especially in group III. In the 3-(4,5-Dimethylthiazol- 2-yl)-2,5-diphenyltetrazolium bromide assay, after 24 hours of incubation, the assessment of cell viability showed that the optical density of the control had a higher tendency than any other group, but there was no significant difference. However, the alkaline phosphatase activity increased as the temperature increased, especially in group III. Conclusions: Consequently, the surface characteristics and biocompatibility increased as the temperature increased. This indicates that surface modification by thermal treatment could be another useful method for medical and dental implants.

Chemical Characteristics and Immunostimulating Activity of Crude Polysaccharide Isolated from Commercial Instant Coffee (시판 인스턴트 커피에서 추출한 다당류의 화학적 특성 및 면역활성)

  • Kwak, Bong-Shin;Shin, Kwang-Soon
    • Korean Journal of Food Science and Technology
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    • v.48 no.3
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    • pp.289-295
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    • 2016
  • To elucidate the new biologically active ingredient in commercial instant coffee, a crude polysaccharide (ICP-0) was isolated by ethanol precipitation, and its immunostimulatory activity was estimated. ICP-0 mainly consisted of galactose (55.5%), mannose (25.7%), arabinose (6.0%), and galacturonic acid (10.1%), suggesting the possibility of its existence as a mixture of galactomannan or pectic polysaccharide. ICP-0 showed proliferative activity in peritoneal macrophages and splenocytes. ICP-0 dose-dependently augmented the production of nitric oxide and reactive oxygen species by peritoneal macrophages. In addition, murine peritoneal macrophages stimulated by ICP-0 showed enhanced production of various cytokines (tumor necrosis factor-${\alpha}$, interleukin-6, and interleukin-12) as compared to unstimulated murine peritoneal macrophages. In an in vitro assay for assessing intestinal immunomodulation, the ICP-0-treated Peyer's patch cells showed higher bone marrow cell proliferation activity at $100{\mu}g/mL$ and higher production of granulocyte-macrophage colony-stimulating factor, compared to the untreated Peyer's patch cells. These results suggest that polysaccharides in commercial instant coffee have a potentiality for macrophage functions and the intestinal immune system.

Effect of Duck's Feet Derived Collagen Sponge on Skin Regeneration: In Vitro Study (오리발 유래 콜라겐 스펀지의 피부재생 효과: In vitro 연구)

  • Cha, Se Rom;Jeong, Hyun Ki;Kim, Su Young;Kim, Eun Young;Song, Jeong Eun;Park, Chan Hum;Kwon, Soon Yong;Khang, Gilson
    • Polymer(Korea)
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    • v.39 no.3
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    • pp.493-498
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    • 2015
  • For biomaterials for skin regeneration with minimized inflammatory response, high bioactivity and biocompatibility are highly required. Also, it should have a porous microstructure to improve cell adhesion and growth. In this study, we extracted a new collagen source from duck's feet which is by-product, and made the shape of sponges from duck's feet collagen (DC) to compare with DBP and SIS. To analyze physical and chemical property of the scaffold, SEM and FTIR were used. MTT assay was used to measure the attachment and proliferation of NIH/3T3 in the scaffolds. RTPCR was used to evaluate the expression of proinflammatory cytokine. Also, 1,1-diphenyl-2-picrylhydrazyl (DPPH) was used to measure the ability of antioxidant activity. Overall, this study shows that DC scaffold is biocompatible and has good physical property. Additionally, DC scaffold shows the potential as wound healing biomaterials.