Cross Protection Against Influenza A Virus by Yeast-Expressed Heterologous Tandem Repeat M2 Extracellular Proteins

The influenza M2 ectodomain (M2e) is well conserved across human influenza A subtypes, but there are few residue changes among avian and swine origin influenza A viruses. We expressed a tandem repeat construct of heterologous M2e sequences (M2e5x) derived from human, swine, and avian origin influenza A viruses using the yeast expression system. Intramuscular immunization of mice with AS04-adjuvanted M2e5x protein vaccines was effective in inducing M2e-specific antibodies reactive to M2e peptide and native M2 proteins on the infected cells with human, swine, or avian influenza virus, mucosal and systemic memory cellular immune responses, and cross-protection against H3N2 virus. Importantly, M2e5x immune sera were found to confer protection against different subtypes of H1N1 and H5N1 influenza A viruses in naïve mice. Also, M2e5x-immune complexes of virusinfected cells stimulated macrophages to secrete cytokines via Fc receptors, indicating a possible mechanism of protection. The present study provides evidence that M2e5x proteins produced in yeast cells could be developed as a potential universal influenza vaccine

Synthesis and Pyrolysis of Novel Polysilazane to SiBCN Ceramic

A novel synthesis route was developed to produce exceptionally thermal stable SiBCN-based ceramics. This route turns out to be cheaper, simpler and faster than the conventional route. The SiBCN-based preceramic polymer was synthesized by using three monomers, trichlorosilane (HSiCl3), boron trichloride (BCl3), and hexamethydisilazane (HMDZ), at a molar ratio of about 1:1:4. The product becomes an amorphous structure with crosslinked bonds during the elimination of (CH3)3SiCl. An intermolecular condensation with the loss of HMDZ occurs in a second stage without the addition of crosslinking agents. In the final pyrolysis stage, hydrogen-rich species such as CH4 are lost. The chemical structure and composition of the obtained polymer were investigated using IR, MAS-NMR, TGA, DSC, XRD, EA, AA, ICP, and EDS. Through the pyrolysis of preceramic polymers, the mechanisms of the structural and compositional changes at a high temperature were also examined. A preceramic polymer useful for nuclear application can be formed, which provides hydrothermal stability under autoclave test

Changes of Shoulder, Elbow, and Wrist Stiffness Matrix Post Stroke

Stroke affects multiple joints in the arm with stereotypical patterns of arm deformity involving the shoulder, elbow, wrist, and hand and with disrupted coordination of multiple joints in active movements. However, there is a lack of systematicmethods to evaluatemulti-joints and multi-degree of freedoms (DOF) neuro-mechanical changes, especially for complex systemswith three ormore joints/ DOFs involved. This paper used a novel systematic method to characterize dynamics and control of the shoulder, elbow, and wrist of the human arm individually and simultaneously, including the couplings across themultiple joints during controlled movements. A novel method was developed to decompose the complex system into manageable single-joint level for more reliable characterizations. The method was used in clinical studies to characterize the multi-joint changes associated with spastic impaired arm of 11 patients post stroke and 12 healthy controls. It was found that stroke survivors showed not only increased stiffness at the individual joints locally but also significantly higher couplings across the joints. The relative increases in couplings are often higher than that of the local joint stiffness. The multi-joint characterization provided a tool to characterize impairment of individual patients, which would allow more focused impairment-specific treatment. In general, the decomposition method can be used for even more complex systems, making characterization of intractable system dynamics of three or more joints/DOFs manageable

Characterization of tandem repeat M2e5x protein.

<p>(A) Structures of M2e5x protein. Human: Human influenza A type M2e, Swine: Swine influenza A type M2e, Avian I: Avian influenza A type I M2e, Avian II: Avian influenza A type II M2e,-: linker. (B) Western blot of M2e5x protein using mouse anti-M2 monoclonal antibody (14C2 mAb). Lane 1: influenza A/Philippines/2/82 (H3N2) virus (40 μg). Lane 2–4: M2e5x protein (10 μg, 5 μg, 2 μg, 1 μg). All samples were run under reduced conditions. (C) SDS-PAGE analysis of M2e5x protein. Lane 1–4: M2e5x protein (40 μg, 20 μg, 10 μg, 5 μg). (D) ELISA reactivity of 14C2 mAb to M2e5x proteins. (E) ELISA reactivity of 14C2 mAb to influenza A/Philippines/2/82 (H3N2) virus.</p

AS04-adjuvanted M2e5x protein vaccination induces M2e-specific IgG and IgG isotype antibody responses.

<p>BALB/c mice (<i>n</i> = 9) were immunized with 20 μg of M2e5x protein alone (M2e5x) or mixed with adjuvant AS04 (M2e5x.AS04). Blood samples were collected at 3 weeks after each immunization. (A) Total IgG antibody. IgG was detected by using human type M2e peptide as an ELISA-coating antigen. (B) M2e-specific IgG isotype responses of the M2e5x.AS04 group after 3rd immunization. (C) IgG antibody responses reactive to M2e peptide antigens derived from human, swine, or avian influenza A viruses. IgG antibodies specific to M2e were measured in 3rd immune sera from the M2e5x.AS04 group using human, swine, avian I, or avian II type peptide as a coating antigen. Sera were serially diluted and ELISA was performed for serum antibodies specific for M2e peptides. Error bars indicates mean ± SEM.</p