High-throughput immunophenotyping by surface plasmon resonance imaging

Cell binding assays on antibody arrays permit the rapid immunophenotyping of living cells. The throughput of the analysis, however, is still limited due to our inability to perform parallel and quantitative detection of cells captured on the array. To address this limitation, we employed here an imaging technique based on surface plasmon resonance (SPR). SPR has been frequently used to monitor capture of proteins on antibody microarrays, while few cases were reported for capture of cells. Antibody arrays were prepared through the photopatterning of an alkanethiol monolayer on a gold-evaporated glass plate and the subsequent immobilization of various antibodies onto 4-9 separate spots created by photopatterning. A glass slip was mounted onto the array with a thin spacer to construct a parallel-plate chamber. Leukemia cells were injected into the chamber to conduct a binding assay, while refractive index changes at the vicinity of the array surface were monitored by SPR imaging. We observed that SPR signals were intensified on specific antibody spots but not on nonspecific spots. Confocal laser scanning microscopy revealed that the observed SPR signals were attributed to cell deformations caused by multivalent interactions with immobilized antibody, which effectively elevated the refractive index of a medium phase within an evanescent field. This effect could be suitably utilized to monitor quantitatively cell binding to multiple spots from a heterogeneous cell population.     

Rewired Cellular Metabolic Profiles in Response to Metformin under Different Oxygen and Nutrient Conditions

Metformin is a metabolic disruptor, and its efficacy and effects on metabolic profiles under different oxygen and nutrient conditions remain unclear. Therefore, the present study examined the effects of metformin on cell growth, the metabolic activities and consumption of glucose, glutamine, and pyruvate, and the intracellular ratio of nicotinamide adenine dinucleotide (NAD⁺) and reduced nicotinamide adenine dinucleotide (NADH) under normoxic (21% O₂) and hypoxic (1% O₂) conditions. The efficacy of metformin with nutrient removal from culture media was also investigated. The results obtained show that the efficacy of metformin was closely associated with cell types and environmental factors. Acute exposure to metformin had no effect on lactate production from glucose, glutamine, or pyruvate, whereas long-term exposure to metformin increased the consumption of glucose and pyruvate and the production of lactate in the culture media of HeLa and HaCaT cells as well as the metabolic activity of glucose. The NAD⁺/NADH ratio decreased during growth with metformin regardless of its efficacy. Furthermore, the inhibitory effects of metformin were enhanced in all cell lines following the removal of glucose or pyruvate from culture media. Collectively, the present results reveal that metformin efficacy may be regulated by oxygen conditions and nutrient availability, and indicate the potential of the metabolic switch induced by metformin as combinational therapy.     

Docosahexaenoic Acid Esters of Hydroxy Fatty Acid Is a Novel Activator of NRF2

Fatty acid esters of hydroxy fatty acids (FAHFAs) are a new class of endogenous lipids with interesting physiological functions in mammals. Despite their structural diversity and links with nuclear factor erythroid 2-related factor 2 (NRF2) biosynthesis, FAHFAs are less explored as NRF2 activators. Herein, we examined for the first time the synthetic docosahexaenoic acid esters of 12-hydroxy stearic acid (12-DHAHSA) or oleic acid (12-DHAHOA) against NRF2 activation in cultured human hepatoma-derived cells (C3A). The effect of DHA-derived FAHFAs on lipid metabolism was explored by the nontargeted lipidomic analysis using liquid chromatography-mass spectrometry. Furthermore, their action on lipid droplet (LD) oxidation was investigated by the fluorescence imaging technique. The DHA-derived FAHFAs showed less cytotoxicity compared to their native fatty acids and activated the NRF2 in a dose-dependent pattern. Treatment of 12-DHAHOA with C3A cells upregulated the cellular triacylglycerol levels by 17-fold compared to the untreated group. Fluorescence imaging analysis also revealed the suppression of the degree of LDs oxidation upon treatment with 12-DHAHSA. Overall, these results suggest that DHA-derived FAHFAs as novel and potent activators of NRF2 with plausible antioxidant function.     

Positive-temperature-coefficient effect of electrical resistivity below melting point of poly(vinylidene fluoride) (PVDF) in Ni particle-dispersed PVDF composites

This paper discusses the positive-temperature-coefficient effects of resistivity in Ni particle-dispersed poly(vinylidene fluoride) (PVDF) composites based on experiment results from SEM, DSC, and pressure-volume-temperature (PVT) measurements. The melting points of composites with Ni content of 20, 30, 40, and 50vol.% were equal to that of pure PVDF. The PTC effects in composites with Ni content of 40 and 50vol.% occurred at temperatures near the melting point of the PVDF matrix, whereas those in composites with Ni content of 20 and 30vol.% occurred at temperatures below the melting point of the PVDF matrix. We found that the PTC effect occurs even without melting of the matrix polymer. Moreover, we determined that a slight increase in specific volume at temperatures below the melting point of the matrix polymer acts fully as a driving force for forming a gap between fillers. This suggestion was backed up by theoretical analyses using percolation theory and a thermal-fluctuation-induced tunneling model.     

Oxidative coupling of methane over LaCoO3−δ-based mixed oxides

The catalytic activity of LaCoO_3–-based mixed oxides for the oxidative coupling of methane has been tested by TPR and cyclic reaction. Characterization has been done by XRD, TGA and Mössbauer spectrometry. It is likely that the perovskite-crystal structure containing hypervalent metal ions has an important role and that unique structural oxygen species in the perovskite contribute to the partial oxidation of methane.     

Surfactant-Assisted Purification of Hydrophobic DNA Nanostructures

Purification of functional DNA nanostructures is an essential step in achieving intended functions because misfolded structures and the remaining free DNA strands in a solution can interact and affect their behavior. However, due to hydrophobicity-mediated aggregation, it is difficult to purify DNA nanostructures modified with hydrophobic molecules by conventional methods. Herein, we report the purification of cholesterol-modified DNA nanostructures by using a novel surfactant-assisted gel extraction. The addition of sodium cholate (SC) to the sample solution before structure folding prevented aggregation; this was confirmed by gel electrophoresis. We also found that adding sodium dodecyl sulfate (SDS) to the sample inhibited structural folding. The cholesterol-modified DNA nanostructures prepared with SC were successfully purified by gel extraction, and their ability to bind to the lipid membrane surfaces was maintained. This method will facilitate the purification of DNA nanostructures modified with hydrophobic molecules and expand their applicability in the construction of artificial cell-like systems.     

Effect of Cu powder addition on thermoelectric properties of Cu/TiO2−x composites

Spark plasma sintering was used to fabricate Cu/TiO_2−x composites by adding Cu powder to nonstoichiometric titanium dioxide, TiO_2−x. The composition and crystal forms of the composites were examined. The thermoelectric properties of the composites were measured and the effects of composite formation on these properties were discussed. The rule of mixture (ROM) of composite and general effective medium theory (GEM) were used to investigate the composite effects of the Cu/TiO_2−x composites. The results revealed that the electrical resistivities of the composites was much lower than that of TiO_2−x. As the added amount of Cu powder increased, the electrical properties of the composites shifted from semiconductor behavior to metallic behavior. The thermoelectric performances of the composites improved as a result of composition formation. The thermoelectric performance can be improved by adjusting the balance among electrical resistivity, thermal conductivity and the Seebeck coefficient, based on the composite effects.     

Surface modification of contact lenses using adsorption of ethylene oxide branched copolymers

To examine methods for reducing the amount of adsorbed protein on the surface of contact lenses during use, cationic copolymers containing poly(ethylene oxide) units were synthesized and evaluated as surface modifiers. Poly(ethylene oxide) graft‐branched copolymers of composition 70 mol % dimethylaminoethyl methacrylate (DM) and 30 mol % methoxy polyethylene glycol methacrylate (Mp0G; p = 2, 4, 9; the average number of the ethylene oxide units) were obtained using nonionic monomers containing poly(ethylene oxide) units. The copolymers very efficiently prevented protein adsorption on a contact lens. Contact angle measurements showed that immersion in tear fluid made the lens surface hydrophobic because of adsorption of proteins with hydrophobic residues. The copolymer pretreatment made the lens surface hydrophilic, even after dipping in artificial tear fluid. These results suggest that adsorption of the poly(ethylene oxide) branched copolymer on the contact lens would make the lens surface hydrophilic and prevent protein adsorption. © 2012 Wiley Periodicals, Inc. J. Appl. Polym. Sci., 2013     

Initial Collection Performance of Resin Wool Filters and Estimation of Charge Density

Abstract

A new type of resin wool filter (RWF) that persists the load with oil droplets was developed by Kimura and colleagues. In the present work, the initial collection performances of RWF (A and C) are measured for various particle sizes (0.03, 0.05, 0.1, 0.15, 0.2, 0.25, and 0.3 μm) with different charging states at various filtration velocities (0.05, 0.1, 0.15, and 0.3 m/s). As a result, it is shown that the present RWF impregnated with PTBP resin can attain high collection efficiency (99.999% at filtration velocity of 0.05 m/s) with a pressure drop of less than 30 Pa. The charge density is estimated by applying prediction equations of single-fiber collection efficiencies of electret filters with a dipolar charge distribution because no other prediction equation for RWF are available at present. The experimental single-fiber efficiencies for uncharged particles are successfully predicted by assigning a single value of charge density in the prediction equations for dipolar fibers. The estimated charge density on RWF fibers is 2.1 × 10^{? 4} C/m², which is much higher than those of conventional electret filter media. Therefore, RWF studied in the present work is suitable for the application to respirators as well as room air cleaners.     

Creation of Titania Artificial Interfaces with Geometric Patterns by Using Microstereolithography and Aqueous Solution Techniques

Titania micropatterns with periodic arrangements were successfully formed on glass substrates for use with electromagnetic wave energy resonance and localizations in terahertz frequency ranges. Geometric arrangements of acrylic polygonal tablets with titania particle dispersions were fabricated by using micropatterning stereolithography. Moreover, periodically arranged full anatase‐phase titania tablets were created homogeneously through liquid‐phase crystal depositions of water solvent processes, using microtemplates fabricated by using the stereolithography system. The terahertz wave properties were measured and calculated by using a time‐domain spectroscopic system and finite‐difference time‐domain method.