• Title/Summary/Keyword: dual-cure

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Soluble Expression of a Human MnSOD and Hirudin Fusion Protein in Escherichia coli, and Its Effects on Metastasis and Invasion of 95-D Cells

  • Yi, Shanze;Niu, Dewei;Bai, Fang;Li, Shuaiguang;Huang, Luyuan;He, Wenyan;Prasad, Anand;Czachor, Alexander;Tan, Lee Charles;Kolliputi, Narasaiah;Wang, Feng
    • Journal of Microbiology and Biotechnology
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    • v.26 no.11
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    • pp.1881-1890
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    • 2016
  • Manganese superoxide dismutase (MnSOD) is a vital enzyme that protects cells from free radicals through eliminating superoxide radicals ($O^{2-}$). Hirudin, a kind of small active peptide molecule, is one of the strongest anticoagulants that can effectively cure thrombus diseases. In this study, we fused Hirudin to the C terminus of human MnSOD with the GGGGS linker to generate a novel dual-feature fusion protein, denoted as hMnSOD-Hirudin. The hMnSOD-Hirudin gene fragment was cloned into the pET15b (SmaI, CIAP) vector, forming a recombinant pET15b-hMnSOD-Hirudin plasmid, and then was transferred into Escherichia coli strain Rosetta-gami for expression. SDS-PAGE was used to detect the fusion protein, which was expected to be about 30 kDa upon IPTG induction. Furthermore, the hMnSOD-Hirudin protein was heavily detected as a soluble form in the supernatant. The purification rate observed after Ni NTA affinity chromatography was above 95%. The hMnSOD-Hirudin protein yield reached 67.25 mg per liter of bacterial culture. The identity of the purified protein was confirmed by western blotting. The hMnSOD-Hirudin protein activity assay evinced that the antioxidation activity of the hMnSOD-Hirudin protein obtained was $2,444.0{\pm}96.0U/mg$, and the anticoagulant activity of the hMnSOD-Hirudin protein was $599.0{\pm}35.0ATU/mg$. In addition, in vitro bioactivity assay showed that the hMnSOD-Hirudin protein had no or little cytotoxicity in H9c2, HK-2, and H9 (human $CD_4{^+}$, T cell) cell lines. Transwell migration assay and invasion assay showed that the hMnSOD-Hirudin protein could suppress human lung cancer 95-D cell metastasis and invasion in vitro.

Effect of surface treatment of FRC-Post on bonding strength to resin cements (FRC-포스트의 표면처리가 레진시멘트와의 접착력에 미치는 영향)

  • Park, Chan-Hyun;Park, Se-Hee;Kim, Jin-Woo;Cho, Kyung-Mo
    • Restorative Dentistry and Endodontics
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    • v.36 no.2
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    • pp.125-131
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    • 2011
  • Objectives: The purpose of this study was to evaluate the effect of surface treatment of FRC-Post on bonding strength to resin cements. Materials and Methods: Pre-surface treated LuxaPost (DMG), Rely-X Fiber Post (3M ESPE) and self adhesive resin cement Rely-X Unicem (3M ESPE), conventional resin cement Rely-X ARC (3M ESPE), and Rely-X Ceramic Primer (3M ESPE) were used. After completing the surface treatments of the posts, posts and resin cement were placed in clear molds and photo-activation was performed. The specimens were sectioned perpendicular to the FRC-Post into 2 mm-thick segments, and push-out strength were measured. The results of bond strength value were statistically analyzed using independent samples t-test and oneway ANOVA with multiple comparisons using Scheffe's test. Results: Silanization of posts affect to the bond strength in LuxaPost, and did not affect in Rely-X Fiber Post. Rely-X ARC showed higher value than Rely-X Unicem. Conclusions: Silanization is needed to enhance the bond strength between LuxaPost and resin cements.

Effect of water storage on the fracture toughness of dental resin cement used for zirconia restoration (수분이 지르코니아 수복물 전용 레진시멘트의 파괴인성에 미치는 영향에 관한 연구)

  • Goo, Bon-Wook;Kim, Sung-Hun;Lee, Jai-Bong;Han, Jung-Suk;Yeo, In-Sung;Ha, Seung-Ryong;Kim, Hee-Kyung
    • The Journal of Korean Academy of Prosthodontics
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    • v.52 no.4
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    • pp.312-316
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    • 2014
  • Purpose: The aim of this study was to compare the fracture toughness of currently available resin cements for zirconia restorations and evaluate the effect of water storage on fracture toughness of those resin cements. Materials and methods: Single-edge notched specimens ($3mm{\times}6mm{\times}25mm$) were prepared from three currently available dual cure resin cements for zirconia restorations (Panavia F 2.0, Clearfil SA luting and Zirconite). Each resin cement was divided into four groups: immersed in distilled water at $37^{\circ}C$ for 1 (Control group), 30, 90, or 180 days (n=5). Specimens were loaded in three point bending at a cross-head speed of 0.1 mm/s. The maximum load at specimen failure was recorded and the fracture toughness ($K_{IC}$) was calculated. Data were analyzed using one-way ANOVA and multiple comparison $Scheff{\acute{e}}$ test (${\alpha}$=.05). Results: In control group, the mean $K_{IC}$ was $3.41{\pm}0.64MN{\cdot}m^{-1.5}$ for Panavia F, 2.0, $3.07{\pm}0.41MN{\cdot}m^{-1.5}$ for Zirconite, $2.58{\pm}0.30MN{\cdot}m^{-1.5}$ for Clearfil SA luting respectively, but statistical analysis revealed no significant difference between them. Although a gradual decrease of $K_{IC}$ in Panavia F 2.0 and gradual increases of KIC in Clearfil SA luting and Zirconite were observed with storage time, there were no significant differences between immersion time for each cement. Conclusion: The resin cements for zirconia restorations exhibit much higher $K_{IC}$ values than conventional resin cements. The fracture toughness of resin cement for zirconia restoration would not be affected by water storage.

A STUDY ON THE BOND STRENGTHS BETWEEN GLASS IONOMER CEMENT BASES AND COMPOSITE RESINS (글래스 아이오노머 이장재와 복합레진간의 결합강도에 관한 연구)

  • Kim, Min-Hee;Kim, Shin;Jeong, Tae-Sung
    • Journal of the korean academy of Pediatric Dentistry
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    • v.26 no.3
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    • pp.520-527
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    • 1999
  • For the purpose of providing some suggestions in selection of filling materials used in 'sandwich technique', the bond strengths between glass ionomer cement bases and composite resins were investigated and compared. For lining materials, 'Vitrebond' and 'Ketac-fil' were used. Using these two as bases, 10 of each following resins were built up on the top ; Z-100 (light curing resin) Clear-fil (chemical curing resin), Bis-core (dual cure resin), Dyract (compomer), therfore 10 specimens of each group and total of 80 specimens were made. After storing specimens in $37^{\circ}C$ deionized water for 24 hours, the shear bond strengths were measured under universal testing machine with 50 kg of full load scale and 1mm/min of cross-head speed and obtained the results as follows : 1. Over Vitrebond base, Z-100 showed the lowest bond strength but the other three did not show any difference in bond strength. 2. Over Ketac-fil base, Clear-fil showed the highest bond strength followed by Dyract, Bis-core, and Z-100 showed the lowest bond strengths. 3. Whereas Clear-fil showed the similar bond strengths on the Vitrebond base as other resins, it showed the highest bond strength on Ketac-fil base, which showed some difference in bond strength by differing GIC bases. 4. The bond strengths between base materials and composite resin showed a stronger resin-dependence tendency in cases with Ketac-fil bases rather than with Vitrebond bases.

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Effects of activators contained in adhesives on dentin bond strengths (상아질 접착제에 포함된 활성제의 사용 유무가 자가중합 복합레진의 상아질에 대한 전단결합강도에 미치는 영향)

  • Kang, Hye-Kyung;Shin, Joo-Hee;Suh, Kyu-Won;Ryu, Jae-Jun
    • The Journal of Korean Academy of Prosthodontics
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    • v.46 no.5
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    • pp.511-519
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    • 2008
  • Statement of problem: An incompatibility between the initiator systems of self-curing composite resins and light-curing adhesives was supposed recently. Purpose: The purpose of the study was to evaluate the influence of activators for self/dual bonding on dentin shear bonding strengths. Material and methods: Fifty human molars were divided into 5 groups. A flat dentin surface was created for each tooth. A self-curing composite resin (Luxacore) was bonded with the following adhesives (n = 10); One-Step, Prime&Bond NT, AdheSE, Prime&Bond NT and AdheSE were also used in combination with activators. Shear bond strengths were measured after 24 hours of water storage. The specimens were loaded in shear in the Instron until failure at a 1 mm/min crosshead speed. Data were compared using one-way ANOVA and Tukey HSD test (${\alpha}$= 0.05). Results: The dentin adhesive systems in order of decreasing median bond strength were One-Step > Prime&Bond NT with activator, AdheSE with activator > Prime&Bond NT, AdheSE. Among adhesives, One-Step had the highest bond strength. Prime&Bond NT with activator had higher bond strengths than Prime&Bond NT that was used alone, and so was AdheSE. Conclusion: Shear bond strengths were increased in Prime&Bond NT and AdheSE when these were used with activators comparing used without activators. But using activators was not effective clinically comparing One-Step.