42 research outputs found

    Photocatalytic production of hydrogen on Ni/NiO/KNbO_3/CdS nanocomposites using visible light

    Get PDF
    The photocatalytic production of H2 from water splitting was demonstrated on Ni/NiO/KNbO3/CdS nanocomposites using visible light irradiation at wavelengths >400 nm in the presence of isopropanol. The inherent photocatalytic activity of bulk-phase CdS was enhanced by combining Q-sized CdS with KNbO3 and Ni deposited on KNbO3. Enhanced activity is most likely due to effective charge separation of photogenerated electrons and holes in CdS that is achieved by electron injection into the conduction band of KNbO3 and the reduced states of niobium (e.g., Nb(IV) and Nb(III)) are shown to contribute to enhanced reactivity in the KNbO3 composites by mediating effective electron transfer to bound protons. We also observed that the efficient attachment of Q-size CdS and the deposition of nickel on the KNbO3 surface increases H2 production rates. Other factors that influence the rate of H2 production including the nature of the electron donors and the solution pH were also determined. The Ni/NiO/KNbO3/CdS nanocomposite system appears to be a promising candidate for possible practical applications including the production of H2 under visible light

    Enhanced efficiency of generating human-induced pluripotent stem cells using Lin28-30Kc19 fusion protein

    Get PDF
    Induced pluripotent stem cells (iPSCs) have intrinsic properties, such as self-renewal ability and pluripotency, which are also shown in embryonic stem cells (ESCs). The challenge of improving the iPSC generation efficiency has been an important issue and there have been many attempts to develop iPSC generation methods. In this research, we added Lin28, known as one of the reprogramming factors, in the form of a soluble recombinant protein from E. coli to improve the efficiency of human iPSC (hiPSC) generation, in respect of alkaline phosphatase (AP)-positive colonies. To deliver Lin28 inside the cells, we generated a soluble Lin28-30Kc19 fusion protein, with 30Kc19 at the C-terminal domain of Lin28. 30Kc19, a silkworm hemolymph-derived protein, was fused due to its cell-penetrating and protein-stabilizing properties. The Lin28-30Kc19 was treated to human dermal fibroblasts (HDFs), in combination with four defined reprogramming factors (Oct4, Sox2, c-Myc, and Klf4). After 14 days of cell culture, we confirmed the generated hiPSCs through AP staining. According to the results, the addition of Lin28-30Kc19 increased the number and size of generated AP-positive hiPSC colonies. Through this research, we anticipate that this recombinant protein would be a valuable material for increasing the efficiency of hiPSC generation and for enhancing the possibility as a substitute of the conventional method

    Development of a CHO cell line for stable production of recombinant antibodies against human MMP9

    Get PDF
    Abstract Background Human matrix metalloproteinase 9 (hMMP9) is a biomarker in several diseases, including cancer, and the need for developing detectors and inhibitors of hMMP9 is increasing. As an antibody against hMMP9 can be selectively bound to hMMP9, the use of anti-MMP9 antibody presents new possibilities to address hMMP9-related diseases. In this study, we aimed to establish a stable Chinese hamster ovary (CHO) cell line for the stable production of antibodies against hMMP9. Results Weconstructed recombinant anti-hMMP9 antibody fragment-expressing genes and transfected these to CHO cells. We chose a single clone, and successfully produced a full-sized antibody against hMMP9 with high purity, sensitivity, and reproducibility. Subsequently, we confirmed the antigen-binding efficiency of the antibody. Conclusions We developed a novel recombinant anti-hMMP9 antibody via a CHO cell-based mammalian expression system, which has a high potential to be used in a broad range of medical and industrial areas

    Soluble expression and stability enhancement of transcription factors using 30Kc19 cell-penetrating protein

    Get PDF
    Transcription factors have been studied as an important drug candidate. Ever since the successful generation of induced pluripotent stem cells (iPSCs), there has been tremendous interest in reprogramming transcription factors. Because of the safety risks involved in a virus-based approach, many researchers have been trying to deliver transcription factors using nonintegrating materials. Thus, delivery of transcription factors produced as recombinant proteins in E. coli was proposed as an alternative method. However, the low level of soluble expression and instability of such recombinant proteins are potential barriers. We engineered a Bombyx mori 30Kc19 protein as a fusion partner for transcription factors to overcome those problems. We have previously reported that 30Kc19 protein can be produced as a soluble form in E. coli and has a cell-penetrating property and a protein-stabilizing effect. Transcription factors fused with 30Kc19 (Oct4-30Kc19, Sox2-30Kc19, c-Myc-30Kc19, L-Myc-30Kc19, and Klf4-30Kc19) were produced as recombinant proteins. Interestingly, Oct4 and L-Myc were expressed as a soluble form by conjugating with 30Kc19 protein, whereas Oct4 alone and L-Myc alone aggregated. The 30Kc19 protein also enhanced the stability of transcription factors both in vitro and in cells. In addition, 30Kc19-conjugated transcription factors showed rapid delivery into cells and transcriptional activity significantly increased. Overall, 30Kc19 protein conjugation simultaneously enhanced soluble expression, stability, and transcriptional activity of transcription factors. We propose that the conjugation with 30Kc19 protein is a novel approach to solve the technical bottleneck of gene regulation using transcription factors.OAIID:RECH_ACHV_DSTSH_NO:T201623709RECH_ACHV_FG:RR00200001ADJUST_YN:EMP_ID:A002014CITE_RATE:3.376FILENAME:2. (2016.04) Soluble expression and stability enhancement of.pdfDEPT_NM:í™”í•™ìƒëŹŒêł”í•™ë¶€EMAIL:[email protected]_YN:YFILEURL:https://srnd.snu.ac.kr/eXrepEIR/fws/file/0df54ee9-e9f1-4612-9d6e-6deaa8197e3e/linkCONFIRM:

    Enhanced anti-cancer effect using MMP-responsive L-asparaginase fused with cell-penetrating 30Kc19 protein

    Get PDF
    As the acute lymphoblastic leukaemia (ALL) develops, expression of L-asparaginase (ASNase) protein is known to decrease. Therefore, deficiency of the ASNase protein would be regarded as one of the significant indications of the ALL. For the treatment of ALL, recombinant ASNase protein derived from bacterial origin is used which causes cytotoxicity by deprivation of Asn. However, short half-life of the protein is an obstacle for medical use. In order to overcome this limit, recombinant ASNase was fused to 30Kc19 with protein-stabilizing and cell-penetrating properties. As the 30Kc19 protein may induce steric hindrance, we further added a PLGLAG linker sequence (LK) between the ASNase and 30Kc19. The treatment of ASNase-LK-30Kc19 fusion protein demonstrated enhanced stability, cell-penetrating property, and anti-cancer activity. Intracellular delivery of both the non-cleaved and cleaved forms of the protein were observed, suggesting that ASNase acted both internally and externally, performing high anti-cancer activity by effective depletion of intracellular Asn. Additionally, ASNase-LK-30Kc19 showed high selectivity towards cancer cells. In terms of the dosage, releasable ASNase from ASNase-LK-30Kc19 reached the same half-maximal inhibitory concentration at a concentration five times lower than non-releasable ASNase-30Kc19. Altogether, the findings suggest that this fusion approach has potential applications in the treatment of ALL

    Measurement of W+W− production in association with one jet in proton–proton collisions at sqrt(s) = 8TeV with the ATLAS detector

    Get PDF
    The production of W boson pairs in association with one jet in pp collisions at View the MathML sources=8 TeV is studied using data corresponding to an integrated luminosity of 20.3 fb−1 collected by the ATLAS detector during 2012 at the CERN Large Hadron Collider. The cross section is measured in a fiducial phase-space region defined by the presence of exactly one electron and one muon, missing transverse momentum and exactly one jet with a transverse momentum above 25 GeV and a pseudorapidity of |η|<4.5|η|<4.5. The leptons are required to have opposite electric charge and to pass transverse momentum and pseudorapidity requirements. The fiducial cross section is found to be View the MathML sourceσWWfid,1-jet=136±6(stat)±14(syst)±3(lumi) fb. In combination with a previous measurement restricted to leptonic final states with no associated jets, the fiducial cross section of WW production with zero or one jet is measured to be View the MathML sourceσWWfid,≀1-jet=511±9(stat)±26(syst)±10(lumi) fb. The ratio of fiducial cross sections in final states with one and zero jets is determined to be 0.36±0.050.36±0.05. Finally, a total cross section extrapolated from the fiducial measurement of WW production with zero or one associated jet is reported. The measurements are compared to theoretical predictions and found in good agreement

    Nrf2 and NF-ÎșB Signaling Pathways Contribute to Porphyra-334-Mediated Inhibition of UVA-Induced Inflammation in Skin Fibroblasts

    No full text
    In this study, we examined the protective effects of porphyra-334 against UVA-irradiated cellular damage and elucidated the underlying mechanisms. Porphyra-334 prevented UVA-induced cell death and exhibited scavenging activities against intracellular oxidative stress induced by UVA irradiation in skin fibroblasts. We found that porphyra-334 significantly reduced the secretion and expression of IL-6 and TNF-α, reduced nuclear expression of Nuclear factor-ÎșB (NF-ÎșB), and sustained NF-E2-related factor 2 (Nrf2) activation. Further mechanism research revealed that porphyra-334 promoted the Nrf2 signaling pathway in UVA-irradiated skin fibroblasts. Our results show that the antioxidant effect of porphyra-334 is due to the direct scavenging of oxidative stress and its inhibitory effects on NF-ÎșB-dependent inflammatory genes, such as IL-6 and TNF-Îș. Therefore, we hypothesize that boosting the Nrf2- NF-ÎșB-dependent response to counteract environmental stress is a promising strategy for the prevention of UVA-related damage

    Nrf2 and NF-ÎșB Signaling Pathways Contribute to Porphyra-334-Mediated Inhibition of UVA-Induced Inflammation in Skin Fibroblasts

    No full text
    In this study, we examined the protective effects of porphyra-334 against UVA-irradiated cellular damage and elucidated the underlying mechanisms. Porphyra-334 prevented UVA-induced cell death and exhibited scavenging activities against intracellular oxidative stress induced by UVA irradiation in skin fibroblasts. We found that porphyra-334 significantly reduced the secretion and expression of IL-6 and TNF-α, reduced nuclear expression of Nuclear factor-ÎșB (NF-ÎșB), and sustained NF-E2-related factor 2 (Nrf2) activation. Further mechanism research revealed that porphyra-334 promoted the Nrf2 signaling pathway in UVA-irradiated skin fibroblasts. Our results show that the antioxidant effect of porphyra-334 is due to the direct scavenging of oxidative stress and its inhibitory effects on NF-ÎșB-dependent inflammatory genes, such as IL-6 and TNF-Îș. Therefore, we hypothesize that boosting the Nrf2- NF-ÎșB-dependent response to counteract environmental stress is a promising strategy for the prevention of UVA-related damage

    Photocatalytic Hydrogen Production with Visible Light over Pt-Interlinked Hybrid Composites of Cubic-Phase and Hexagonal-Phase CdS

    No full text
    A hybrid photocatalytic system, which is based on a mixed-phase cadmium sulfide matrix composed of nanoparticulate cubic-phase CdS (c-CdS) with average particle diameters of 13 nm and a bandgap energy of 2.6 eV, is coupled with bulk-phase hexagonal CdS (hex-CdS) that has a bandgap energy of 2.3 eV and is interlinked with elemental platinum deposits. The resulting hybrid nanocomposite catalysts are photocatalytically efficient with respect to hydrogen gas production from water with visible light irradiation at λ > 420 nm. Rates of H_2 production approaching 1.0 mmol-H_2 g^(−1) h^(−1) are obtained with a c-CdS/Pt/hex-CdS composite photocatalyst, in the presence of a mixed sodium sulfide and sodium sulfite background electrolyte system at pH 14. In contrast, the same composite produces H_2 a rate of 0.15 mmol g^(−1) h^(−1) at pH 7 in a water-isopropanol solvent system. The relative order of reactivity for the synthesized hybrid catalysts was found to be c-CdS/Pt/hex-CdS > Pt/c-CdS/hex-CdS > Pt/hex-CdS > hex-CdS > c-CdS/hex-CdS > quantum-sized c-CdS. A mechanism involving enhanced lifetimes of electron−hole trapping states that are dependent on the surface chemistry of hydrated CdS involving surface hydroxyl (>CdOH) and sulfhydryl groups (>CdSH) are invoked
    corecore