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  • Calcium hydroxide regulates bone sialoprotein gene transcription in human osteoblast-like Saos2 cells. 21467818

    Bone sialoprotein (BSP) is a mineralized tissue-specific protein expressed in differentiated osteoblasts that appears to function in the initial mineralization of bone. Calcium hydroxide (Ca(OH)(2)) is a basic salt that has been widely used for a variety of applications in dentistry, due to its antimicrobial effects and its capability of inducing hard tissue formation. However, details of the mechanism involved in the mineralization induced by Ca(OH)(2) are still unclear. In the present study, Ca(OH)(2) (0.4 mM) was found to increase the levels of BSP and Runx2 mRNA at 3 h in human osteoblast-like Saos2 cells. Transient transfection assays were performed using chimeric constructs of the human BSP gene promoter linked to a luciferase reporter gene. Treatment of Saos2 cells with Ca(OH)(2) (0.4 mM) increased the luciferase activities of the constructs between -60LUC and -927LUC at 12 h. Gel shift analysis showed that Ca(OH)(2) (0.4 mM) increased the binding of nuclear protein to CRE1, CRE2 and FRE. Antibodies against CREB1, c-Fos, c-Jun, JunD, Fra2 and P300 disrupted the formation of the CRE1- and CRE2-protein complexes, and antibodies against Dlx5, Msx2, Runx2 and Smad1 disrupted the formation of the FRE-protein complex. These findings demonstrate that Ca(OH)(2) stimulates BSP transcription by targeting the CRE1, CRE2 and FRE elements in the human BSP gene promoter.
    Document Type:
    Reference
    Product Catalog Number:
    AB5728

  • Aluminum hydroxide adjuvants activate caspase-1 and induce IL-1beta and IL-18 release. 17404311

    Aluminum hydroxide (Alum) is the only adjuvant approved for routine use in humans, although the basis for its adjuvanticity remains poorly understood. In this study, we show that Alum activates caspase-1 and induce secretion of mature IL-1beta and IL-18. Human PBMC or dendritic cells stimulated with pure TLR4 and TLR2 agonists released only traces of IL-1beta or IL-18, despite the fact that the IL-1beta mRNA was readily induced by both TLR agonists. In contrast, cells costimulated with TLR agonists plus Alum released large amount of IL-1beta and IL-18. Alum-induced IL-1beta and IL-18 production was not due to enhancement of TLR signaling but rather reflected caspase-1 activation and in mouse dendritic cells occurred in a MyD88-independent fashion. Secretion of other proinflammatory cytokines such as IL-8 was not affected by Alum treatments. However, TLR-induced production of IL-10 was increased and that of IFN-gamma-inducible protein decreased by Alum cotreatment. Considering the immunostimulatory activities of these cytokines and the ability of IL-1beta to act as adjuvant, our results suggest a mechanism for the adjuvanticity of Alum.
    Document Type:
    Reference
    Product Catalog Number:
    06-503
    Product Catalog Name:
    Anti-Caspase 1 Antibody

  • Recombinant viral-like particles of parvovirus B19 as antigen carriers of anthrax protective antigen. 16724664

    Viral-like particles (VLPs) of parvovirus B19 were employed as an antigen carrier to present antigenic determinants of Bacillus anthracis. The small-loop peptide and the full-length domain 4 of protective antigen (PA) were chosen as immunogens for presentation on the VLP-capsid surface and subsequent immunization of BALB/c mice. The recombinant VLPs induced anti-PA IgG titers of up to 2.5 x 10(4). Neutralization assays showed that the recombinant VLPs elicited neutralizing anti-PA antibody titers of up to 1:400 and showed potential for the prevention of lethal toxin-induced mortality of mouse-macrophage cells (RAW264.7). In postimmune sera, no anti-PA titers were detected against synthetic small-loop peptide. Recombinant VLPs demonstrated the capacity to retain the immunogenicity of the displayed microbial PA-epitopes and elicited robust levels of anti-PA antibody titers. These findings suggest that the recombinant VLPs of parvovirus B19 have potential as an additional tool in the development of sub-unit vaccines.
    Document Type:
    Reference
    Product Catalog Number:
    MAB8293
    Product Catalog Name:
    Anti-Parvovirus B19 Antibody, aa 328-344 of VP2 capsid protein, clone R92F6

  • Proteomic analyses of corneal tissue subjected to alkali exposure. 20861482

    To determine whether exposure to alkaline chemicals results in predictable changes in corneal protein profile. To determine whether protein profile changes are indicative of severity and duration of alkali exposure.Enucleated bovine and porcine (n = 59 each) eyes were used for exposure to sodium, ammonium, and calcium hydroxide, respectively. Eyes were subjected to fluorescein staining, 5-bromo-2'-deoxy-uridine (BrdU) labeling. Excised cornea was subjected to protein extraction, spectrophotometric determination of protein amount, dynamic light scattering and SDS-PAGE profiling, mass spectrometric protein identification, and iTRAQ-labeled quantification. Select identified proteins were subjected to Western blot and immunohistochemical analyses.Alkali exposure resulted in lower protein extractability from corneal tissue. Elevated aggregate formation was found with strong alkali exposure (sodium hydroxidegreater than ammonium, calcium hydroxide), even with a short duration of exposure compared with controls. The protein yield after exposure varied as a function of postexposure time. Protein profiles changed because of alkali exposure. Concentration and strength of the alkali affected the profile change significantly. Mass spectrometry identified 15 proteins from different bands with relative quantification. Plexin D1 was identified for the first time in the cornea at a protein level that was further confirmed by Western blot and immunohistochemical analyses.Exposure to alkaline chemicals results in predictable and reproducible changes in corneal protein profile. Stronger alkali, longer durations, or both, of exposure resulted in lower yields and significant protein profile changes compared with controls.
    Document Type:
    Reference
    Product Catalog Number:
    MAB374
    Product Catalog Name:
    Anti-Glyceraldehyde-3-Phosphate Dehydrogenase Antibody, clone 6C5

  • An organic matrix-mediated processing methodology to fabricate hydroxyapatite based nanostructured biocomposites. 20644842

    An amorphous calcium phosphate precursor phase, which forms by adding orthophosphoric acid to a calcium hydroxide suspension, is transformed into crystalline hydroxyapatite by introducing polymer solutions. The nanostructured composite films formed by a solvent casting technique from the concentrated hybrid suspension are characterised for structure and mechanical properties.
    Document Type:
    Reference
    Product Catalog Number:
    09-432
    Product Catalog Name:
    Anti-phospho-IRS1 (Tyr608) mouse/ (Tyr612) human Antibody

  • Preparation of chitosan films using different neutralizing solutions to improve endothelial cell compatibility. 22042456

    The development of chitosan-based constructs for application in large-size defects or highly vascularized tissues is still a challenging issue. The poor endothelial cell compatibility of chitosan hinders the colonization of vascular endothelial cells in the chitosan-based constructs, and retards the establishment of a functional microvascular network following implantation. The aim of the present study is to prepare chitosan films with different neutralization methods to improve their endothelial cell compatibility. Chitosan salt films were neutralized with either sodium hydroxide (NaOH) aqueous solution, NaOH ethanol solution, or ethanol solution without NaOH. The physicochemical properties and endothelial cell compatibility of the chitosan films were investigated. Results indicated that neutralization with different solutions affected the surface chemistry, swelling ratio, crystalline conformation, nanotopography, and mechanical properties of the chitosan films. The NaOH ethanol solution-neutralized chitosan film (Chi-NaOH/EtOH film) displayed a nanofiber-dominant surface, while the NaOH aqueous solution-neutralized film (Chi-NaOH/H(2)O film) and the ethanol solution-neutralized film (Chi-EtOH film) displayed nanoparticle-dominant surfaces. Moreover, the Chi-NaOH/EtOH films exhibited a higher stiffness as compared to the Chi-NaOH/H(2)O and Chi-EtOH films. Endothelial cell compatibility of the chitosan films was evaluated with a human microvascular endothelial cell line, HMEC-1. Compared with the Chi-NaOH/H(2)O and Chi-EtOH films, HMECs cultured on the Chi-NaOH/EtOH films fully spread and exhibited significantly higher levels of adhesion and proliferation, with retention of the endothelial phenotype and function. Our findings suggest that the surface nanotopography and mechanical properties contribute to determining the endothelial cell compatibility of chitosan films. The nature of the neutralizing solutions can affect the physicochemical properties and endothelial cell compatibility of chitosan films. Therefore, selection of suitable neutralization methods is highly important for the application of chitosan in tissue engineering.
    Document Type:
    Reference
    Product Catalog Number:
    MAB2150
    Product Catalog Name:
    Anti-E-Selectin Antibody, clone P2H3