IL-12 Production Induced by Agaricus blazei Fraction H (ABH) Involves Toll-like Receptor (TLR)

Agaricus blazei Murill is an edible fungus used in traditional medicine, which has various well-documented medicinal properties. In the present study, we investigated the effects of hemicellulase-derived mycelia extract (Agaricus blazei fraction H: ABH) on the immune system. First, we examined the cytokine-inducing activity of ABH on human peripheral mononuclear cells (PBMC). The results indicated that ABH induced expression of IL-12, a cytokine known to be a critical regulator of cellular immune responses. Flow cytometric analysis demonstrated the induction of IL-12 production by the CD14-positive cell population, consisting of monocytes/macrophages (Mo/Mφ). Furthermore, the elimination of Mo/Mφ attenuated IL-12 production in PBMC. ABH-induced IL-12 production was inhibited by anti-CD14 and anti-TLR4 antibodies but not by anti-TLR2 antibody. The activity of ABH was not inhibited by polymyxin B, while the activity of lipopolysaccharide used as a reference was inhibited. Oral administration of ABH enhanced natural killer (NK) activity in the spleen. These findings suggest that ABH activated Mo/Mφ in a manner dependent on CD14/TLR4 and NK activity

The role of homophilic binding in anti-tumor antibody R24 recognition of molecular surfaces. Demonstration of an intermolecular beta-sheet interaction between vh domains.

The murine antibody R24 and mouse-human Fv-IgG1(kappa) chimeric antibody chR24 are specific for the cell-surface tumor antigen disialoganglioside GD3. X-ray diffraction and surface plasmon resonance experiments have been employed to study the mechanism of "homophilic binding," in which molecules of R24 recognize and bind to other molecules of R24 though their heavy chain variable domains. R24 exhibits strong binding to liposomes containing disialoganglioside GD3; however, the kinetics are unusual in that saturation of binding is not observed. The binding of chR24 to GD3-bearing liposomes is significantly weaker, suggesting that cooperative interactions involving antibody constant regions contribute to R24 binding of membrane-bound GD3. The crystal structures of the Fabs from R24 and chR24 reveal the mechanism for homophilic binding and confirm that the homophilic and antigen-binding idiotopes are distinct. The homophilic binding idiotope is formed largely by an anti-parallel beta-sheet dimerization between the H2 complementarity determining region (CDR) loops of two Fabs, while the antigen-binding idiotope is a pocket formed by the three CDR loops on the heavy chain. The formation of homophilic dimers requires the presence of a canonical conformation for the H2 CDR in conjunction with participation of side chains. The relative positions of the homophilic and antigen-binding sites allows for a lattice of GD3-specific antibodies to be constructed, which is stabilized by the presence of the cell membrane. This model provides for the selective recognition by R24 of cells that overexpress GD3 on the cell surface

A mutational analysis of the globotriaosylceramide-binding sites of verotoxin VT1.

Escherichia coli verotoxin, also known as Shiga-like toxin, binds to eukaryotic cell membranes via the glycolipid Gb(3) receptors which present the P(k) trisaccharide Galalpha(1-4)Galbeta(1-4)Glcbeta. Crystallographic studies have identified three P(k) trisaccharide (P(k)-glycoside) binding sites per verotoxin 1B subunit (VT1B) monomer while NMR studies have identified binding of P(k)-glycoside only at site 2. To understand the basis for this difference, we studied binding of wild type VT1B and VT1B mutants, defective at one or more of the three sites, to P(k)-glycoside and pentavalent P(k) trisaccharide (pentaSTARFISH) in solution and Gb(3) presented on liposomal membranes using surface plasmon resonance. Site 2 was the key site in terms of free trisaccharide binding since mutants altered at sites 1 and 3 bound this ligand with wild type affinity. However, effective binding of the pentaSTARFISH molecule also required a functional site 3, suggesting that site 3 promotes pentavalent binding of linked trisaccharides at site 1 and site 2. Optimal binding to membrane-associated Gb(3) involved all three sites. Binding of all single site mutants to liposomal Gb(3) was weaker than wild type VT1B binding. Site 3 mutants behaved as if they had reduced ability to enter into high avidity interactions with Gb(3) in the membrane context. Double mutants at site 1/site 3 and site 2/site 3 were completely inactive in terms of binding to liposomal Gb(3,) even though the site 1/site 3 mutant bound trisaccharide with almost wild type affinity. Thus site 2 alone is not sufficient to confer high avidity binding to membrane-localized Gb(3). Cytotoxic activity paralleled membrane glycolipid binding. Our data show that the interaction of verotoxin with the Gb(3) trisaccharide is highly context dependent and that a membrane environment is required for biologically relevant studies of the interaction

Accurate large-signal equivalent circuit of surface channel diamond FETs based on the Chalmers model

The paper presents a large-signal nonlinear circuit-oriented model for polycrystalline and single-crystal H-terminated diamond MESFETs implemented within the Agilent ADS design suite. The DC characteristics of such devices suggest that the channel free charge control law may be modeled using the same strategy adopted for III-V HEMTs. For this reason, the well-known nonlinear Chalmers (Angelov) circuit model was chosen as the starting point for the development of the present non-linear diamond MESFET model. Model fitting was performed against DC and multibias small signal measurements, with good agreement. Model validations versus large-signal (power) measurements point out the accuracy of the proposed approach to simulate the behavior of H-terminated diamond MESFETs under large-signal operatio

Diamond-Based Thin Film Bulk Acoustic Wave Resonator for Biomedical Applications

Nowadays it is in constant growing the development of thin film bulk acoustic resonators. If the piezoelectric material is going to be implanted in the human body, an important requirement is the biocompatibility of the implant. In this regard, Aluminum Nitride (AlN) has emerged as an attractive alternative for use in biomedical MicroElectroMechanical Systems. Ultrananocrystalline Diamond (UNCD) is a promising material to be used in biomedical applications, due to its extraordinary mulifunctionality; it is exceptional for implantable medical devices requiring stringent biological performance. Since both UNCD and AlN films can be processed via photolithography processes used in microfabrication, the integration of UNCD and AlN films provides the bases for developing a new generation of biocompatible Bio-MEMS/NEMS. Research and development was conducted to produce implantable MEMS devices: Pt/piezoelectric AlN/Pt layer heterostructure was grown and patterned on the UNCD membrane with a Ti adhesion layer. By applying voltages between the top and bottom Pt electrodes layers the piezoelectric AlN layer is energized. The feasibility of the fabrication of biocompatible AlN/diamond-based FBAR structure has been demonstrated.Fil: Zalazar, Martin. Universidad Nacional de Entre Rios. Facultad de Ingenieria. Departamento de Bioingenieria; ArgentinaFil: Guarnieri, Fabio Ariel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico. Centro de Investigación de Métodos Computacionales; Argentina. Universidad Nacional de Entre Rios. Facultad de Ingenieria. Departamento de Bioingenieria; Argentin

A short sequence for the iterative synthesis of fused polyethers

A simple and efficient four‐step sequence for the synthesis of fused polyether arrays has been developed. Cyclic ethers are installed by sequential alkynyl ether formation, carbocupration, ring‐closing metathesis and hydroboration with acidic workup. Crucially, the alkene required for the subsequent ring formation by ring‐closing metathesis is present in the substrate but is masked in the form of a vinylic silane, which prevents competitive metathesis of the side chain. Generation of the reactive alkene from the unreactive vinylic silane is accomplished by hydroboration and subsequent acid‐mediated Peterson elimination of the intermediate hydroxysilane