Simple replication methods for producing nanoslits in thermoplastics and the transport dynamics of double-stranded DNA through these slits

Mixed-scale nano- and microfluidic networks were fabricated in thermoplastics using simple and robust methods that did not require the use of sophisticated equipment to produce the nanostructures. High-precision micromilling (HPMM) and photolithography were used to generate mixed-scale molding tools that were subsequently used for producing fluidic networks into thermoplastics such as poly(methyl methacrylate), PMMA, cyclic olefin copolymer, COC, and polycarbonate, PC. Nanoslit arrays were imprinted into the polymer using a nanoimprinting tool, which was composed of an optical mask with patterns that were 2-7 μm in width and a depth defined by the Cr layer (100 nm), which was deposited onto glass. The device also contained a microchannel network that was hot embossed into the polymer substrate using a metal molding tool prepared via HPMM. The mixed-scale device could also be used as a master to produce a polymer stamp, which was made from polydimethylsiloxane, PDMS, and used to generate the mixed-scale fluidic network in a single step. Thermal fusion bonding of the cover plate to the substrate at a temperature below their respective T(g) was accomplished by oxygen plasma treatment of both the substrate and cover plate, which significantly reduced thermally induced structural deformation during assembly: ～6% for PMMA and ～9% for COC nanoslits. The electrokinetic transport properties of double-stranded DNA (dsDNA) through the polymeric nanoslits (PMMA and COC) were carried out. In these polymer devices, the dsDNA demonstrated a field-dependent electrophoretic mobility with intermittent transport dynamics. DNA mobilities were found to be 8.2 ± 0.7 × 10(-4) cm(2) V(-1) s(-1) and 7.6 ± 0.6 × 10(-4) cm(2) V(-1) s(-1) for PMMA and COC, respectively, at a field strength of 25 V cm(-1). The extension factors for λ-DNA were 0.46 in PMMA and 0.53 in COC for the nanoslits (2-6% standard deviation).     

Fractionation and characterization of gold nanoparticles in aqueous solution: asymmetric-flow field flow fractionation with MALS, DLS, and UV–Vis detection

Asymmetrical-flow field flow fractionation (AFFF) separates constituents based on hydrodynamic size and is emerging as a powerful tool for obtaining high-resolution information on the size, molecular weight, composition, and stability of nanoscale particles in liquid media. We employ a customized AFFF system combining on-line detectors for multi-angle light scattering, dynamic light scattering, and UV–Vis absorption. Our objective is to develop optimized measurement protocols for the characterization of gold nanoparticles (GNPs), which are widely utilized in biomedical research and other nanotechnology applications. Experimental conditions have been optimized by controlling key parameters, including injection volume and solids concentration, mobile phase composition, membrane type and pore size, and ratio of channel-to-cross-flow rates. Individual citrate-stabilized GNP components (nominally 10, 20, 30, 40, and 60 nm) and GNPs functionalized with polyethylene glycol were separated from multicomponent GNP mixtures by AFFF and subsequently characterized. We discuss the effects due to variations in measurement parameters and GNP surface modification on observed retention, recovery, and peak resolution.     

Three-dimensional optical microscopy using axially distributed image sensing

We propose three-dimensional (3D) optical microscopy using axially distributed image sensing. In the proposed method, the micro-objects are optically magnified and their axially distributed images are recorded by moving the image sensor along a common optical axis. The 3D volumetric images are generated from the recorded axial image set using a computational reconstruction algorithm based on ray backprojection. Preliminary experimental results are presented. To the best of our knowledge, this is the first report on 3D optical microscopy using axially distributed sensing.     

An Oxidizable Anion‐Excluding Polymer‐Overlayer for Oxygen Electrodes

Here we describe an anion excluding ion‐permeable membrane, which we evaluate on an O₂‐electroreducing cathode poised at a strongly oxidizing potential, near the reversible potential of the O₂/H₂O half cell. The bioelectrocatalyst of the O₂ cathode consists of the cross‐linked electrostatic adduct of a polycationic redox hydrogel and bilirubin oxidase (BOD), a polyanion at neutral at pH 7.3. If an uncured Nafion dispersion is applied on this bioelectrocatalyst, the polyanionic Nafion displaces the BOD in the electrostatic adduct, de‐wiring the BOD. We show here that insertion of a polycationic poly(acrylamide‐co‐vinylimidazole) (PAA‐PVI) between the bioelectrocatalyst and the Nafion prevents the dewiring of BOD. The resulting bi‐layer membrane effectively excludes the urate, thiocyanate and NADH anions.     

Selective addition of CXCR3+CCR4-CD4+ Th1 cells enhances generation of cytotoxic T cells by dendritic cells in vitro

Increasing importance is being given to the stimulation of Th1 response in cancer immunotherapy because its presence can shift the direction of adaptive immune responses toward protective immunity. Based on chemokine receptor expression, CXCR3⁺CCR4^-CD4⁺ T cells as Th1-type cells were investigated its capacity in monocyte-derived dendritic cell (DC) maturation and polarization, and induction of antigen specific cytotoxic T lymphocytes (CTL) in vitro. The levels of IL-4, IL-5 and IL-10 were decreased to the basal level compared with high production of IFN-γ, TNF-α, and IL-2 in CXCR3⁺CCR4^-CD4⁺ T cells stimulated with anti-CD3 and anti-CD28 antibodies. Co-incubation of activated CD4⁺ or CXCR3⁺CCR4^-CD4⁺ T cells with DC (CD4⁺/DC or CXCR3⁺CD4⁺/DC, respectively) particularly up-regulated IL-12 and CD80 expression compared with DC matured with TNF-α and LPS (mDC). Although there was no significant difference between the effects of the CXCR3⁺CCR4^-CD4⁺ and CD4⁺ T cells on DC phenotype expression, CXCR3⁺CD4⁺/DC in CTL culture were able to expand number of CD8⁺ T cells and increased frequencies of IFN-γ secreting cells and overall cytolytic activity against tumor antigen WT-1. These results demonstrated that the selective addition of CXCR3⁺CCR4^-CD4⁺ T cells to CTL cultures could enhance the induction of CTLs by DC in vitro, and implicated on a novel strategy for adoptive T cell therapy.     

Alterations of epinephrine-induced gluconeogenesis in aging

The effects of glucagon and epinephrine on gluconeogenesis in young (4 month) and old (24 month) Fisher 344 rat hepatocytes were compared. In contrast to glucagon, which had a similar effect on gluconeogenesis in both young and old cells, epinephrine caused a smaller increase in gluconeogenesis in old rat hepatocytes than in young hepatocytes. β₂ adrenergic receptor (β₂-AR) expression slightly decreased in aged rat liver, and there were differences between young and old hepatocytes in their patterns of G protein coupled receptor kinases, which are involved in the activation of β₂-AR receptor signal desensitization. The major isoform of the kinase changed from GRK2 to GRK3 and the expression of β-arrestin, which is recruited by the phosphorylated β₂-AR for internalization and degradation, increased in aged rat liver. GRK3 overexpression also decreased the glucose output from young rat hepatocytes. We conclude that an age-associated reduction in epinephrine-induced gluconeogenesis occurs through the epinephrine receptor desensitizing system.     

XPS/EXAFS study of cycleability improved LiMn2O4 thin film cathodes prepared by solution deposition

The influence of Sn substitution in LiMn₂O₄ thin films as a cathode has been studied via solution deposition to improve the electrochemical performance of thin film lithium batteries. LiSn_0.025Mn_1.95O₄ thin films showed the most promising performance, i.e. a high capacity retention of 77% at 10 C after the 500th cycle, due to the increased average Mn valence state. The thin films of LiSn_x/2Mn_2?xO₄ (x ? 0.10) showed significant precipitation of SnO₂ and SnO after the cycling evaluation.     

Self‐Template‐Directed Formation of Coordination‐Polymer Hexagonal Tubes and Rings,and their Calcination to ZnO Rings

This template will self‐destruct : A unique particle‐growth mechanism involves growth of new coordination polymers on the surface of initially formed hexagonal blocks and concomitant dissolution of the blocks to form hexagonal tubes (see scheme and scanning electron, optical, and fluorescence microscopy images). Calcination of the tubes yields ZnO particles of the same shape.

     

HPLC of fluoroquinolone antibacterials using chiral stationary phase based on enantiomeric (3,3′‐diphenyl‐1,1′‐binaphthyl)‐20‐crown‐6

A residual silanol group‐protecting chiral stationary phase (CSP) based on optically active (3,3′‐diphenyl‐1,1′‐binaphthyl)‐20‐crown‐6 was successfully applied to the resolution of fluoroquinolone compounds including gemifloxacin mesylate. The chiral recognition ability of the residual silanol group‐protecting CSP was generally greater than that of the residual silanol group‐containing CSP. From these results, it was concluded that the simple protection of the residual silanol groups of the latter CSP with lipophilic n‐octyl groups can improve its chiral recognition ability for the resolution of racemic fluoroquinolone compounds. The chromatographic resolution behaviors were investigated as a function of the content and type of organic and acidic modifiers and the ammonium acetate concentration in aqueous mobile phase and the column temperature. Especially, the addition of ammonium acetate to the mobile phase was found to be a quite effective means of reducing the enantiomer retentions without sacrificing the chiral recognition efficiency of the CSP.     

A simple formula for an analogue of conditional wiener integrals and its applications II

Let C[0, T] denote the space of real-valued continuous functions on the interval [0, T] with an analogue w _ϕ of Wiener measure and for a partition 0 = t ₀ < t ₁ < ... < t _n < t _n+1 = T of [0, T], let X _n : C[0, T] → ℝ ⁿ⁺¹ and X ⁿ⁺¹: C[0, T] → ℝ ⁿ⁺² be given by X _n (x) = (x(t ₀), x(t ₁), ..., x(t _n )) and X _n+1(x) = (x(t ₀), x(t ₁), ..., x(t _n+1)), respectively. In this paper, using a simple formula for the conditional w _ϕ-integral of functions on C[0, T] with the conditioning function X _n+1, we derive a simple formula for the conditional w _ϕ-integral of the functions with the conditioning function X _n . As applications of the formula with the function X _n , we evaluate the conditional w _ϕ-integral of the functions of the form F _m (x) = ∫₀ ^T (x(t)) ^m for x ∈ C[0, T] and for any positive integer m. Moreover, with the conditioning X _n , we evaluate the conditional w _ϕ-integral of the functions in a Banach algebra which is an analogue of the Cameron and Storvick’s Banach algebra . Finally, we derive the conditional analytic Feynman w _ϕ-integrals of the functions in .