Alloy design concept for bcc-T2 silicide-B2 aluminide multicomponent alloys

For the development of refractory metal-based high temperature bcc alloys, the phase equilibrium between bcc (Nb-Mo) and T2 (Nb, Mo)5(Si,B)3 has been investigated. Bcc matrix phase is for toughening at ambient temperatures, and T2 phase is for strengthening and also for oxidation resistance. However, the oxidation resistance of T2 phase is still under investigation. B2-NiAl phase has been utilized as coating materials for Ni-based superalloys for many years. However, addition of Al and transition metal element such as Ni and Fe results in the formation of brittle Laves phases in refractory metal-based bcc alloys. In the present study it is found that additive element selection in terms of atomic size control is effective to avoid the formation of Laves phase. From this phase stability viewpoint, a three-phase alloy composed of Nb, Mo, Si, B, Ni and Al is proposed as a first step for designing three-phase alloys. Please click Additional Files below to see the full abstract

Immunobiotic Bifidobacteria Strains Modulate Rotavirus Immune Response in Porcine Intestinal Epitheliocytes via Pattern Recognition Receptor Signaling.

In this work, we aimed to characterize the antiviral response of an originally established porcine intestinal epithelial cell line (PIE cells) by evaluating the molecular innate immune response to rotavirus (RVs). In addition, we aimed to select immunomodulatory bacteria with antiviral capabilities. PIE cells were inoculated with RVs isolated from different host species and the infective titers and the molecular innate immune response were evaluated. In addition, the protection against RVs infection and the modulation of immune response by different lactic acid bacteria (LAB) strains was studied. The RVs strains OSU (porcine) and UK (bovine) effectively infected PIE cells. Our results also showed that RVs infection in PIE cells triggered TLR3-, RIG-I- and MDA-5-mediated immune responses with activation of IRF3 and NF-κB, induction of IFN-β and up-regulation of the interferon stimulated genes MxA and RNase L. Among the LAB strains tested, Bifidobacterium infantis MCC12 and B. breve MCC1274 significantly reduced RVs titers in infected PIE cells. The beneficial effects of both bifidobacteria were associated with reduction of A20 expression, and improvements of IRF-3 activation, IFN-β production, and MxA and RNase L expressions. These results indicate the value of PIE cells for studying RVs molecular innate immune response in pigs and for the selection of beneficial bacteria with antiviral capabilities

Innate immune response of porcine intestinal epithelial cell line (PIE cells) after infection with rotaviruses.

<p>PIE cells were cultured in DMEM media for 10 days, and then infected with OSU or UK rotaviruses. After 0, 3, 6, and 12 hours post-infection the expression level of IFN-β, MxA, RNaseL, RIG-I, TLR3, and cytokines (CXCL10, IL-6, IL-8, and MCP-1) were quantified. The results represent data from three independent experiments and are expressed as mean ± S.D. Asterisks indicate significant differences: <i>p</i> <0.05 (*), <i>p</i> <0.01 (**), and <i>p</i> < 0.001(***).</p

Effect of immunobiotic bifidobacteria on IRF3 activation in porcine intestinal epithelial cell line (PIE cells).

<p>PIE cells were pre-treated with <i>B</i>. <i>infantis</i> MCC12 or <i>B</i>. <i>breve</i> MCC1274, and then challenged with rotaviruses OSU or UK; or poly(I:C). Total proteins were extracted from lysed cells and separated by SDS-PAGE, and then western-blot was performed to analyze phosphorylation of IRF3 after 10, 30, 60 and 120 minutes. The phosphorylated IRF3 specific bands were normalized to that corresponding to total IRF3. Intensities of proteins bands were calculated from peak area of densitogram by using image software. The results represent data from three independent experiments and are expressed as relative index vs 0 min control with mean ± S.D. ^{a, b, c} Different superscripts letters indicate significant difference (p<0.05) among stimulants at the same time point.</p

Effect of immunobiotic bifidobacteria on TRAF3 activation in porcine intestinal epithelial cell line (PIE cells).

<p>PIE cells were pre-treated with <i>B</i>. <i>infantis</i> MCC12 or <i>B</i>. <i>breve</i> MCC1274, and then challenged with rotaviruses OSU or UK; or poly(I:C). Total proteins were extracted from lysed cells and separated by SDS-PAGE, and then western-blot was performed to analyze phosphorylation of TRAF3 after 10, 30, 60 and 120 minutes. The TRAF3 specific bands were normalized to that corresponding to β-actin. Intensities of proteins bands were calculated from peak area of densitogram by using image software. The results represent data from three independent experiments and are expressed as relative index vs 0 min control with mean ± S.D. ^{a, b, c} Different superscripts letters indicate significant difference (p<0.05) among stimulants at the same time point.</p

Infectivity of rotaviruses in porcine intestinal epithelial cell line (PIE cells).

<p>PIE cells cultured for 3 or 10 days. MA104 cells were used for comparison. PIE and MA104 cells were inoculated with porcine OSU or bovine UK rotaviruses and evaluated by immunofluorescence assay. The cells with specific green-fluorescence in the cytoplasm were photographed by confocal laser microscopy after labeling with fluorescence antibody.</p

Infectivity of rotaviruses in porcine intestinal epithelial cell line (PIE cells).

<p>PIE cells cultured for 3 or 10 days. MA104 cells were used for comparison. PIE and MA104 cells were inoculated with rotaviruses from different host species including: human Wa, murine EW, bovine UK, and porcine OSU rotaviruses. Numbers of rotavirus antigen positive cells were counted by immunofluorescence assay after 16 hours post-inoculation. The virus titer was expressed as Log₁₀ focus forming units (FFU)/0.1 ml. NI: no infectivity of rotaviruses. The results represent data from three independent experiments. ^{a, b, c} In the comparison within means of the same virus, the difference among means with different superscripts was significant at 5% level.</p

Proposed mechanism.

<p>The possible immunomodulatory activity of <i>B</i>. <i>infantis</i> MCC12, and <i>B</i>. <i>breve</i> MCC1274 in porcine intestinal epithelial cell line (PIE cells) after stimulation with rotaviruses. Arrows indicate up and down-regulation of cytokines/chemokines, and anti-viral factors. (+): upregulation, (--): strong down-regulation, (-): moderate down-regulation.</p

Effect of immunobiotic bifidobacteria in antiviral immune response of porcine intestinal epithelial cell line (PIE cells).

<p>PIE cells were pre-treated with <i>B</i>. <i>infantis</i> MCC12 or <i>B</i>. <i>breve</i> MCC1274 for 48 hours, and then challenged with rotavirus UK. The expression level of IFN-β, MxA, RNaseL, RIG-I, TLR3, A20, and cytokines (CXCL10, IL-6, IL-8, MCP-1) were quantified by RT-PCR after 6 and 12 hours. The results represent data from three independent experiments and are expressed as mean ± S.D. Asterisks indicate significant differences: <i>p</i> <0.05 (*), <i>p</i> <0.01 (**), and <i>p</i> < 0.001(***).</p