SSA-MOA: a novel CTC isolation platform using selective size amplification (SSA) and a multi-obstacle architecture (MOA) filter

Circulating tumor cells (CTCs) have gained increasing attention as physicians and scientists learn more about the role these extraordinarily rare cells play in metastatic cancer. In developing CTC technology, the critical criteria are high recovery rates and high purity. Current isolation methods suffer from an inherent trade-off between these two goals. Moreover, ensuring minimal cell stress and robust reproducibility is also important for the clinical application of CTCs. In this paper, we introduce a novel CTC isolation technology using selective size amplification (SSA) for target cells and a multi-obstacle architecture (MOA) filter to overcome this trade-off, improving both recovery rate and purity. We also demonstrate SSA-MOA's advantages in minimizing cell deformation during filter transit, resulting in more stable and robust CTC isolation. In this technique, polymer microbeads conjugated with anti-epithelial cell adhesion molecules (anti-EpCAM) were used to selectively size-amplify MCF-7 breast cancer cells, definitively differentiating from the white blood cells (WBCs) by avoiding the size overlap that compromises other size selection methods. 3 μm was determined to be the optimal microbead diameter, not only for size discrimination but also in maximizing CTC surface coverage. A multi-obstacle architecture filter was fabricated using silicon-on-glass (SOG) technology-a first such application of this fabrication technique-to create a precise microfilter structure with a high aspect ratio. The filter was designed to minimize cell deformation as simulation results predicted that cells captured via this MOA filter would experience 22% less moving force than with a single-obstacle architecture. This was verified by experiments, as we observed reliable cell capture and reduced cell deformation, with a 92% average recovery rate and 351 peripheral blood leukocytes (PBL) per millilitre (average). We expect the SSA-MOA platform to optimize CTC recovery rates, purity, and stability, increasing the sensitivity and reliability of such tests, thereby potentially expanding the utilization of CTC technologies in the clinic.     

Magnetically-actuated blood filter unit attachable to pre-made biochips

We present a novel blood filter unit that is designed to separate blood plasma from whole blood by simple magnetic actuation. A non-diluted blood sample is dropped into the filter unit and magnetic attraction is applied to squeeze out only blood plasma while blood particles are filtered by membranes stacked in the filter unit. The new filter device yields good filtering performance with nearly perfect filtering efficiency (～99.999%), high plasma recovery (～30%), low blood consumption (<50 μl), and fast operation (～1 min). Because it is simple to operate and is attachable to any kind of pre-made biochip, it has commercial potential in various lab-on-a-chip applications for blood tests.     

Transition state analysis of Vibrio cholerae sialidase-catalyzed hydrolyses of natural substrate analogues

A series of isotopically labeled natural substrate analogues (phenyl 5-N-acetyl-α-d-neuraminyl-(2→3)-β-d-galactopyranosyl-(1→4)-1-thio-β-d-glucopyranoside; Neu5Acα2,3LacβSPh, and the corresponding 2→6 isomer) were prepared chemoenzymatically in order to characterize, by use of multiple kinetic isotope effect (KIE) measurements, the glycosylation transition states for Vibrio cholerae sialidase-catalyzed hydrolysis reactions. The derived KIEs for Neu5Acα2,3LacβSPh for the ring oxygen ((18)V/K), leaving group oxygen ((18)V/K), C3-S deuterium ((D)V/K(S)) and C3-R deuterium ((D)V/K(R)) are 1.029 ± 0.002, 0.983 ± 0.001, 1.034 ± 0.002, and 1.043 ± 0.002, respectively. In addition, the KIEs for Neu5Acα2,6βSPh for C3-S deuterium ((D)V/K(S)) and C3-R deuterium ((D)V/K(R)) are 1.021 ± 0.001 and 1.049 ± 0.001, respectively. The glycosylation transition state structures for both Neu5Acα2,3LacβSPh and Neu5Acα2,6LacβSPh were modeled computationally using the experimental KIE values as goodness of fit criteria. Both transition states are late with largely cleaved glycosidic bonds coupled to pyranosyl ring flattening ((4)H(5) half-chair conformation) with little or no nucleophilic involvement of the enzymatic tyrosine residue. Notably, the transition state for the catalyzed hydrolysis of Neu5Acα2,6βSPh appears to incorporate a lesser degree of general-acid catalysis, relative to the 2,3-isomer.     

Understanding the mechanism of short-range electron transfer using an immobilized cupredoxin

The hydrophobic patch of azurin (AZ) from Pseudomonas aeruginosa is an important recognition surface for electron transfer (ET) reactions. The influence of changing the size of this region, by mutating the C-terminal copper-binding loop, on the ET reactivity of AZ adsorbed on gold electrodes modified with alkanethiol self-assembled monolayers (SAMs) has been studied. The distance-dependence of ET kinetics measured by cyclic voltammetry using SAMs of variable chain length, demonstrates that the activation barrier for short-range ET is dominated by the dynamics of molecular rearrangements accompanying ET at the AZ-SAM interface. These include internal electric field-dependent low-amplitude protein motions and the reorganization of interfacial water molecules, but not protein reorientation. Interfacial molecular dynamics also control the kinetics of short-range ET for electrostatically and covalently immobilized cytochrome c. This mechanism therefore may be utilized for short-distance ET irrespective of the type of metal center, the surface electrostatic potential, and the nature of the protein-SAM interaction.     

Development of Zn–ProPhenol‐Catalyzed Asymmetric Alkyne Addition: Synthesis of Chiral Propargylic Alcohols

The development of a general and practical zinc‐catalyzed enantioselective alkyne addition methodology is reported. The commercially available ProPhenol ligand ( 1 ) has facilitated the addition of a wide range of zinc alkynylides to aryl, aliphatic, and α,β‐unsaturated aldehydes in high yield and enantioselectivity. New insights into the mechanism of this reaction have resulted in a significant reduction in reagent stoichiometry, enabling the use of precious alkynes and avoiding the use of excess dimethylzinc. The enantioenriched propargylic alcohols from this reaction serve as versatile synthetic intermediates and have enabled efficient syntheses of several complex natural products.     

Influence of High-energy electron-beam on photocatalytic activity of TiO2 films on carbon-fiber deposited by atomic layer deposition

High-energy electron-beam with energy of 1 MeV was used for modifying surface structure of TiO₂ thin films on carbon fiber prepared by using atomic layer deposition under atmospheric pressure. TiO₂ nanoparticles (∼20 nm) on carbon fiber underwent structural modification of the surface upon electron-beam treatment, resulting in enhanced photocatalytic activity. In contrast, a thicker film of TiO₂ did not show such changes in surface structure and photocatalytic activity by electron-beam treatment. We demonstrate that electron-beam can be used for modifying surface structure of photocatalysts consisting of nanoparticles for improvement of their activity.     

Stokes phenomena and non-perturbative completion in the multi-cut two-matrix models

The Stokes multipliers in the matrix models are invariants in the string-theory moduli space and related to the D-instanton chemical potentials. They not only represent non-perturbative information but also play an important role in connecting various perturbative string theories in the moduli space. They are a key concept to the non-perturbative completion of string theory and also expected to imply some remnant of strong coupling dynamics in M theory. In this paper, we investigate the non-perturbative completion problem consisting of two constraints on the Stokes multipliers. As the first constraint, Stokes phenomena which realize the multi-cut geometry are studied in the Zk symmetric critical points of the multi-cut two-matrix models. Sequence of solutions to the constraints are obtained in general k-cut critical points. A discrete set of solutions and a continuum set of solutions are explicitly shown, and they can be classified by several constrained configurations of the Young diagram. As the second constraint, we discuss non-perturbative stability of backgrounds in terms of the Riemann-Hilbert problem. In particular, our procedure in the 2-cut (1,2) case (pure-supergravity case) completely fixes the D-instanton chemical potentials and results in the Hastings-McLeod solution to the Painlevé II equation. It is also stressed that the Riemann-Hilbert approach realizes an off-shell background independent formulation of non-critical string theory.     

Multichannel transceiver dual-tuned RF coil for proton/sodium MR imaging of knee cartilage at 3 T

Sodium magnetic resonance (MR) imaging is a promising technique for detecting changes of proteoglycan (PG) content in cartilage associated with knee osteoarthritis. Despite its potential clinical benefit, sodium MR imaging in vivo is challenging because of intrinsically low sodium concentration and low MR signal sensitivity. Some of the challenges in sodium MR imaging may be eliminated by the use of a high-sensitivity radiofrequency (RF) coil, specifically, a dual-tuned (DT) proton/sodium RF coil which facilitates the co-registration of sodium and proton MR images and the evaluation of both physiochemical and structural properties of knee cartilage. Nevertheless, implementation of a DT proton/sodium RF coil is technically difficult because of the coupling effect between the coil elements (particularly at high field) and the required compact design with improved coil sensitivity. In this study, we applied a multitransceiver RF coil design to develop a DT proton/sodium coil for knee cartilage imaging at 3 T. With the new design, the size of the coil was minimized, and a high signal-to-noise ratio (SNR) was achieved. DT coil exhibited high levels of reflection S11 (～-21 dB) and transmission coefficient S12 (～-19 dB) for both the proton and sodium coils. High SNR (range 27-38) and contrast-to-noise ratio (CNR) (range 15-21) were achieved in sodium MR imaging of knee cartilage in vivo at 3-mm(3) isotropic resolution. This DT coil performance was comparable to that measured using a sodium-only birdcage coil (SNR of 28 and CNR of 20). Clinical evaluation of the DT coil on four normal subjects demonstrated a consistent acquisition of high-resolution proton images and measurement of relative sodium concentrations of knee cartilages without repositioning of the subjects during the same MR scanning session.     

Pure spin current generation in monolayer graphene by quantum pumping

We study a method to generate pure spin current in monolayer graphene over a wide range of Fermi energy by adiabatic quantum pumping. The device consists of three gate electrodes and two ferromagnetic strips, which induce a spin-splitting in the graphene through the proximity effect. A pure spin current is generated by applying two periodic oscillating gate voltages. We find that the pumped pure spin current is a sensitive oscillatory function of the Fermi energy. Large spin currents can be found at Fermi energies where there are Fabry-Perot resonances in the barriers. Furthermore, we analyze the effects of the parameters of the system on the pumped currents. Our predicted pumped spin current can be of the order of 100 nA which is measurable using the current technology. The proposed method is useful in the realization of graphene spintronic devices.     

复杂混合溶液成分高光谱分析的可行性

采用高光谱技术对复杂混合溶液进行检测分析,同时利用被测物质的吸光度和散射特性信息以提高光谱的信噪比。实验设计了高光谱采集装置,采集生物组织模拟液(Intralipid-10%)的漫反射高光谱图像,并用Monte Carlo方法和漫射近似理论对其进行了正向和反向推导,获得了632nm波长下,Intralipid-10%吸收系数为0.002 0cm-1,与标准参数相对误差为11.1%;约化散射系数为63.35cm-1,与标准参数相对误差为6.49%,基本符合标准参数的误差范围,验证了该高光谱检测系统的准确性。还利用该高光谱系统对不同厂家出品的牛奶、果汁等样本进行了高光谱采集,得到不同样本间差异较传统二维光谱更为明显的结果,充分证明了高光谱方法在复杂混合溶液成分分析中具有很强的可行性。