• Title/Summary/Keyword: Ascorbate

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Isolation of High-Quality mRNA from Tannin-Rich Persimmon Fruit (고 Tannin 함유 감과실로 부터 mRNA의 분리)

  • ;Dav
    • Food Science and Preservation
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    • v.4 no.1
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    • pp.45-51
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    • 1997
  • In our studies on the role of $\beta$-galactosidase in fruit softening, significant difficulty, was encountered in our attempts to extract RNA from persimmon(Diospyros kaki L. cv. Fuyu) fruit due to astringency and tannin content. Initial, unsuccessful RNA extractions involved methods using guanidinium isothiocyanate/CsCl with and without polyvinylpyrrolidone(PVP), phenol/sodium lauryl sulfate(SDS), guanidinium hydrochloride, as well as polysomal RNA purification method that used 0.2 M Tris-HCI (pH 9.0) containing KCI, Mg-acetate, EDTA, $\beta$-mercaptoethanol, and sucrose. A method was devised which employed treatment of fruit with CO2 gas to diminish astringency prior to RNA extraction, followed by extraction of tissue powders with Proteinase K extraction buffer containing PVP and ascorbate at an alkaline pH. This procedure resulted in the removal of tannins and other polyphenolics and extraction of relatively large amount of high-quality RNA suitable for cDNA library construction and polymerase chain reaction(PCR). Futhermore, the procedure does not use the toxic and corrosive chemical guanidinium isothiocyanate or require ultracentrifugation.

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Effective application duration of sodium ascorbate antioxidant in reducing microleakage of bonded composite restoration in intracoronally-bleached teeth

  • Park, Jae-Young;Kwon, Tae-Yub;Kim, Young-Kyung
    • Restorative Dentistry and Endodontics
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    • v.38 no.1
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    • pp.43-47
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    • 2013
  • Objectives: The aim of this study was to determine an appropriate application duration of sodium ascorbate (SA) antioxidant gel in reducing microleakage of bonded composite restoration in intracoronally-bleached teeth. Materials and Methods: Eighty endodontically-treated human incisors were randomly divided into eight groups: control, no bleaching; IB and DB, immediate and delayed bonding after bleaching, respectively; S10m, S60m, S24h, S3d and S7d, bleaching + SA gel for 10 min, 60 min, 24 hr, 3 day and 7 day, respectively. For bleaching, a mixture of 30% hydrogen peroxide and sodium perborate was applied for 7 day. All access cavities were restored using One-Step adhesive (Bisco Inc.) and then Aelite LS Packable composite (Bisco Inc.). The bonded specimens were subjected to 500 thermal cycles, immersed in 1% methylene blue for 8 hr, and longitudinally sectioned. Microleakage was assessed with a 0 - 4 scoring system and analyzed using nonparametric statistical methods (${\alpha}$ = 0.05). Results: Group IB showed a significantly higher microleakage than the control group (p = 0.006) and group DB a statistically similar score to the control group (p > 0.999). Although groups S10m, S60m, and S24h exhibited significantly higher scores than group DB (p < 0.05), the microleakage in groups S3d and S7d was statistically similar to that in group DB (p = 0.771, p > 0.999). Conclusions: Application of SA gel for 3 day after nonvital bleaching was effective in reducing microleakage of composite restoration in intracoronally-bleached teeth.

Gene Gun-Mediated Human Erythropoietin Gene Expression in Primary Cultured Oviduct Cells from Laying Hens

  • Ochiai, H.;Park, H.M.;Sasaki, R.;Okumura, J.;Muramatsu, T.
    • Asian-Australasian Journal of Animal Sciences
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    • v.12 no.1
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    • pp.9-14
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    • 1999
  • Factors affecting gene gun-mediated expression of the human erythropoietin gene were investigated in primary cultured oviduct cells from laying hens. The human erythropoietin gene was transfected by a gene gun method at $1.25{\mu}g$ per dish, and cultured in a synthetic serum-free medium for 72 hrs. The concentration of human erythropoietin mRNA was determined by RNA : RNA solution hybridization. In experiment 1, the effect of changing the shooting pressure of DNA-coated microparticles with nitrogen gas was tested at 20 and $60kgf/cm^2$. The results showed that the erythropoietin mRNA concentration was significantly higher at 60 than $20kgf/cm^2$. In experiment 2, the effects of supplementing the medium with fetal calf serum at 10%, and raising the shooting pressure from 60 to $80kgf/cm^2$ on the cell number and erythropoietin gene expression were examined. Although supplementation with fetal calf serum significantly increased the cell numbes compared with no supplemented controls (p < 0.05), erythropoietin mRNA concentration per $10^3$ cells was not affected. Raising the shooting pressure from 60 to $80kgf/cm^2$ did not affect either of the parameters, In experiment 3, the effect of supplementing ascorbate 2-phosphate at 0.5 mM was tested. The results indicated that the ascorbate supplementation significantly increased the cell number (p < 0.05), and tended to increase erythropoietin mRNA concentration (p < 0.1). Thus, for human erythropoietin gene expression by using the gene gun method, shooting pressure with nitrogen gas should be sufficient at $60kgf/cm^2$ and supplementation with ascorbate phosphate would be useful to enhance not only the cell proliferation but also erythropoietin gene expression.

The Relationshin between ACE Inhibitory Activity and Degradations of Sulfur Containing Materials in Dolsan Leaf Mustard Juice

  • Yoo Eun-Jeong;Choi Myeong-Rak;Lim Hyun-Soo
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.9 no.5
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    • pp.400-404
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    • 2004
  • This Study was tarried out to investigate the relationship ACE inhibitory activity and degradations of sulfur containing materials in Dolsan leaf mustard juice (DLMJ). The changes of sulfur containing materials which were treated with autolysis, myrosinase, ascorbate and papain were studied, as well as the changes of ACE inhibitory activity in DLMJ. At $37^{\circ}C$, sulfur contain-ing materials by autolysis decreased most rapidly from $0.43\%$ to $0.13\%$ in the second day. Conversely. ACE inhibitory activity increased most from $66\%$ to $87\%$. in the second day at $37^{\circ}C$. As myrosinase concentrations increased more, sulfur containing materials in DLMJ decreased more. The ACE inhibitory activities at 0, 0.5, 1, 2, and 4 Units of myrosinase for 240 min later were 70, 74, 75, 82, and $85\%$, respectively. At 1 mM ascorbate. concentrations of Sulfur containing materials in DLMJ decreased more significantly on the second day than on the other days. At 1 mM ascorbate for 6 days, ACE Inhibitory activity reached a maximum of about $92\%$. And, an increase of papain concentration was noted in accordance with a decreased sulfur containing materials. The maximum rate of AEC inhibitory activity at control, 3, 6, and 12 Units of papains treatments was shown as 70, 70, 75, and $78\%$ at 60 min, respectively. These results suggested that the degradation of sulfur containing materials led to the increase of ACE inhibitory activity. Consequently, it was suggested that ACE inhibiting was significantly related to the degradatives of sulfur containing materials.