Protective Immunity against Listeria monocytogenes in Rats,Provided by HCl- and NaOH-Induced Listeria monocytogenes Bacterial Ghosts (LMGs) as Vaccine Candidates

Listeria monocytogenes (Lm) bacterial ghosts (LMGs) were produced by the minimum inhibitory concentration (MIC) of HCl, H₂SO₄, and NaOH. Acid and alkali effects on the LMGs were compared by in vitro and in vivo analyses. Scanning electron microscope showed that all chemicals form lysis pores on the Lm cell envelopes. Real-time qPCR revealed a complete absence of genomic DNA in HCl- and H₂SO₄-induced LMGs but not in NaOH-induced LMGs. HCl-, H₂SO₄- and NaOH-induced LMGs showed weaker or missing protein bands on SDS-PAGE gel when compared to wild-type Lm. Murine macrophages exposed to the HCl-induced LMGs showed higher cell viability than those exposed to NaOH-induced LMGs or wild-type Lm. The maximum level of cytokine expression (TNF-α, iNOS, IFN-γ, and IL-10 mRNA) was observed in the macrophages exposed to NaOH-induced LMGs, while that of IL-1β mRNA was observed in the macrophages exposed to HCl-induced LMGs. To investigate LMGs as a vaccine candidate, mice were divided into PBS buffer-injected, HCl- and NaOH-induced LMGs immunized groups. Mice vaccinated with HCl- and NOH-induced LMGs, respectively, significantly increased in specific IgG antibodies, bactericidal activities of serum, and CD4⁺ and CD8⁺ T-cell population. Antigenic Lm proteins reacted with antisera against HCl- and NOH-induced LMGs, respectively. Bacterial loads in HCl- and NaOH-induced LMGs immunized mice were significantly lower than PBS-injected mice after virulent Lm challenges. It suggested that vaccination with LMGs induces both humoral and cell-mediated immune responses and protects against virulent challenges.     

Rhabdomyolysis-Induced AKI Was Ameliorated in NLRP3 KO Mice via Alleviation of Mitochondrial Lipid Peroxidation in Renal Tubular Cells

Introduction: A recent study showed that early renal tubular injury is ameliorated in Nod-like receptor pyrin domain-containing protein 3 (NLRP3) KO mice with rhabdomyolysis-induced acute kidney injury (RIAKI). However, the precise mechanism has not been determined. Therefore, we investigated the role of NLRP3 in renal tubular cells in RIAKI. Methods: Glycerol-mediated RIAKI was induced in NLRP3 KO and wild-type (WT) mice. The mice were euthanized 24 h after glycerol injection, and both kidneys and plasma were collected. HKC-8 cells were treated with ferrous myoglobin to mimic a rhabdomyolytic environment. Results: Glycerol injection led to increase serum creatinine, aspartate aminotransferase (AST), and renal kidney injury molecule-1 (KIM-1) level; renal tubular necrosis; and apoptosis. Renal injury was attenuated in NLRP3 KO mice, while muscle damage and renal neutrophil recruitment did not differ between NLRP3 KO mice and WT mice. Following glycerin injection, increases in cleaved caspase-3, poly (ADP-ribose) polymerase (PARP), and a decrease in the glutathione peroxidase 4 (GPX-4) level were observed in the kidneys of mice with RIAKI, and these changes were alleviated in the kidneys of NLRP3 KO mice. NLRP3 was upregulated, and cell viability was suppressed in HKC-8 cells treated with ferrous myoglobin. Myoglobin-induced apoptosis and lipid peroxidation were significantly decreased in siNLRP3-treated HKC-8 cells compared to ferrous myoglobin-treated HKC-8 cells. Myoglobin reduced the mitochondrial membrane potential and increased mitochondrial fission and reactive oxygen species (ROS) and lipid peroxidation levels, which were restored to normal levels in NLRP3-depleted HKC-8 cells. Conclusions: NLRP3 depletion ameliorated renal tubular injury in a murine glycerol-induced acute kidney injury (AKI) model. A lack of NLRP3 improved tubular cell viability via attenuation of myoglobin-induced mitochondrial injury and lipid peroxidation, which might be the critical factor in protecting the kidney.     

Replacing Mg2+ by Fe2+ for RNA-Cleaving DNAzymes

It has been proposed that Mg²⁺ and Fe²⁺ are very similar in interacting with ribozymes and some protein-based enzymes, but their activities with DNAzymes have yet to be studied. Here, the activity of Fe²⁺ as cofactor for a few RNA-cleaving DNAzymes is investigated. 17E is a well-studied DNAzyme that is active in the presence of many different divalent metal ions; it is highly active with Fe²⁺ with an apparent K_d of 29.7±2.3 μm and a k_obs of 1.12±0.11 min⁻¹ in the presence of 1 mm Fe²⁺ at pH 7.5. Fe²⁺ has 21-fold higher activity than Mg²⁺. Six different DNAzymes are then tested, and only the DNAzymes active with Mg²⁺ (17E, 8–17, and E5) are active with Fe²⁺. Fe²⁺ has 25 and one- to twofold higher activity than Mg²⁺ for the 8–17 and E5 DNAzymes, respectively. In pH>7 buffer and in presence of air, 1 mm Fe²⁺ results in a nonspecific degradation of the DNA strand due to reactive oxygen species (ROS). Cleavage reactions in anoxic environment and antioxidant ascorbate can be used to overcome the effect of oxidation. The findings provide insights for potential DNAzyme catalysis in the early Earth, and they further support the similarity between Mg²⁺ and Fe²⁺ in enzyme catalysis.     

Dietary Monounsaturated Fatty Acids but not Saturated Fatty Acids Preserve the Insulin Signaling Pathway via IRS-1/PI3K in Rat Skeletal Muscle

Saturated fatty acids (SFA) and monounsaturated fatty acids (MUFA) show different effects on the development of insulin resistance. In this study, we compared the effect of dietary SFA and MUFA on the insulin signaling pathway in the skeletal muscle of a type 2 diabetic animal model. Twenty-nine-week-old male Otsuka Long-Evans Tokushima fatty (OLETF) rats were randomly divided into three groups and fed one of the following diets for 3 weeks; a normal chow diet, an SFA (lard oil) enriched or a MUFA (olive oil) enriched high-fat diet. The vastus lateralis muscle was used for analyses. Insulin tolerance test showed improved insulin sensitivity in rats fed the MUFA diet, as compared to those fed the SFA diet (p < 0.001). The SFA diet reduced IRS-1 expression and phosphorylated PI3K levels in skeletal muscle, as compared with a chow diet (p < 0.001, respectively). On the contrary, muscle IRS-2 expression and phosphorylated ERK1/2 was significantly increased in rats fed the SFA diet (p < 0.001, respectively). Membrane translocation of glucose transporter type 4 decreased in the skeletal muscle of rats fed the SFA diet, as compared to those fed a chow diet (p < 0.001). These changes in insulin signaling pathway in skeletal muscle were not observed in rats fed the MUFA diet. In conclusion, the beneficial effect of dietary MUFA on insulin sensitivity is associated with a conserved IRS-1/PI3K insulin signaling pathway which was altered by dietary SFA.     

Epoxy resins toughened with in situ azide–alkyne polymerized polysulfones

To simultaneously improve the fracture toughness and heat resistance of a cured toughened epoxy resin along with a reduction in its viscosity during the mixing process, two novel polysulfone‐type polymers are synthesized via azide–alkyne polymerization for use as toughening agents. The epoxy resin toughened with these polymers by in situ azide–alkyne polymerization during the cure process, which shows excellent processibility and based on the significantly lower viscosity (61 and 62 cP) during epoxy mixing process than that of commonly commercial polyethersulfone (PES, 127,612 cP). The novel polysulfone‐type polymer toughened epoxy resin showed the advantage in excellent fracture toughness than the PES toughened epoxy. In addition, the glass transition temperature of the novel polysulfone‐type polymer toughened epoxy resin is similar to that of the neat one (～230 °C) and does not decrease, which implies excellent heat resistance of the toughened epoxy. These phenomena can be attributed to the formation of semi‐interpenetrating polymer networks comprising the epoxy network and the linear polysulfone‐type polymers. © 2017 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2018 , 135, 45790.     

Particle size effect of Li[Ni0.5Mn0.5]O2 prepared by co-precipitation

Spherical (Ni_0.5Mn_0.5)(OH)₂ with different secondary particle size (3 μm, 10 μm in diameter) was synthesized by co-precipitation method. Mixture of the prepared hydroxide and lithium hydroxide was calcined at 950 °C for 20 h in air. X-ray diffraction patterns revealed that the prepared material had a typical layered structure with space group. Spherical morphologies with mono-dispersed powders were observed by scanning electron microscopy. It was found that the layered Li[Ni_0.5Mn_0.5]O₂ delivered an initial discharge capacity of 148 mAh g⁻¹ (3.0-4.3 V) though the particle sizes were different. Li[Ni_0.5Mn_0.5]O₂ having smaller particle size (3 μm) showed improved area specific impedance due to the reduced Li⁺ diffusion path, compared with that of Li[Ni_0.5Mn_0.5]O₂ possessing larger particle size (10 μm). Although the Li[Ni_0.5Mn_0.5]O₂ (3 μm) was electrochemically delithiated to Li_0.39[Ni_0.5Mn_0.5]O₂, the resulting exothermic onset temperature was around 295 °C, of which the value is significantly higher than that of highly delithiated Li_1−δCoO₂ (∼180 °C).     

Steam reforming of n-hexadecane over noble metal-modified Ni-based catalysts

Steam reforming of n-hexadecane, a main constituent of diesel, over noble metal-modified Ni-based hydrotalcite catalyst was carried out in a temperature range of 700–950 °C, at an atmospheric pressure with space velocity of 10,000–100,000 h⁻¹ and feed molar ratio of H₂O/C = 3.0. The catalysts were prepared by a co-precipitation and dipping methods. The noble metal-modified Ni-based hydrotalcite catalyst displayed higher resistance for the sintering of active metal than the Ni-based hydrotalcite catalyst prepared by the conventional method. It was found that the Rh-modified Ni-based catalysts showed high resistance to the formation of carbon compared to Ni-based catalysts. The results suggest that Rh-modified Ni-based catalyst can be applied for the steam reforming (SR) reaction of diesel.     

Immobilization of pamidronic acids on the nanotube surface of titanium discs and their interaction with bone cells

Self-assembled layers of vertically aligned titanium nanotubes were fabricated on a Ti disc by anodization. Pamidronic acids (PDAs) were then immobilized on the nanotube surface to improve osseointegration. Wide-angle X-ray diffraction, X-ray photoelectron microscopy, and scanning electron microscopy were employed to characterize the structure and morphology of the PDA-immobilized TiO₂ nanotubes. The in vitro behavior of osteoblast and osteoclast cells cultured on an unmodified and surface-modified Ti disc was examined in terms of cell adhesion, proliferation, and differentiation. Osteoblast adhesion, proliferation, and differentiation were improved substantially by the topography of the TiO₂ nanotubes, producing an interlocked cell structure. PDA immobilized on the TiO₂ nanotube surface suppressed the viability of the osteoclasts and reduced their bone resorption activity.     

Development and characterization of covalently cross-linked SPEEK/Cs-TPA/CeO2 composite membrane and membrane electrode assembly for water electrolysis

Covalently cross-linked SPEEK/Cs-TPA/CeO₂ composite membrane was prepared for the polymer electrolyte membrane water electrolysis. Tungstophosphoric acid (TPA) with a cesium was added to the SPEEK to increase proton conductivity. CeO₂ was used to scavenge free radicals which attack the membrane in the water electrolysis and to improve the durability of the membrane. The composite membrane featured the electrochemical characteristics, such as 0.130 S/cm of proton conductivity at 80 °C, and 2.324 meq./g-dry-memb. of ion-exchange capacity. Pt(NH₃)₄Cl₂, Pd(NH₃)₄Cl₂, RhCl₃ and Co(NH₆)₄Cl₃ were used to prepare a variety of the membrane electrode assemblies (MEAs) as electrocatalytic precursors. Electrochemical activity surface area (ESA) of the Pt–Pd electrode prepared with 2 mM Pt(NH₃)₄Cl₂ and 2 mM Pd(NH₃)₄Cl₂ showed the best properties of 26.2 m²/g with CL-SPEEK/Cs-TPA/CeO₂ membrane. In water electrolysis performance, the cell voltage of Pd/PEM/Pt–Pd MEA with CL-SPEEK/Cs-TPA/CeO₂(1%) composite membrane showed cell property of 1.82 V at 1 A cm⁻² and 80 °C.     

Characteristics of gas-liquid chromatography

The effects of sample size, liquid loading, particle size, column length, and column temperature on retention volumes were studied and separation factor, column efficiency, partition coefficient, and heat of solution were also obtained by gas-liquid chromatography. The feed materials were chosen by similar boiling points as diethylether, dimethoxymethane and dichloromethane. The relations between retention volume and above mentioned various variables were obtained. Separation factor was more affected by column temperature than other variables, and decreased with the temperature. HETP increased almost linearly with sample size. From the exponential relationship between partition coefficient and column temperature heat of solution of each material was calculated.