Detection of six single-nucleotide polymorphisms associated with rheumatoid arthritis by a loop-mediated isothermal amplification method and an electrochemical DNA chip

An electrochemical DNA chip using an electrochemically active intercalator and DNA probe immobilized on a gold electrode has been developed for genetic analysis. In this study, the six polymorphisms associated with rheumatoid arthritis (RA), N-acetyltransferase2 (NAT2) gene polymorphisms T341C, G590A, and G857A, methylenetetrahydrofolate reductase (MTHFR) gene polymorphisms C677T and A1298C, and serum amyloid A1 (SAA1) gene promoter polymorphism C-13T were simultaneously detected by the electrochemical DNA chip and the loop-mediated isothermal amplification (LAMP) method, which is a novel technique for DNA amplification. Human genomic DNAs were extracted from blood, and the targets containing the six polymorphisms were amplified by the LAMP method. A sample containing the six LAMP products was reacted with the electrochemical DNA chip using a DNA detection system that controls hybridization reaction, washing, electrochemical detection, and data analysis automatically. A total of 31 samples were genotyped by this method, and the results were completely consistent with those determined by the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis or the PCR direct sequence analysis. The time required for this method was only 2 h, and operations were very simple. Therefore, this method is expected to contribute to personalized medicine based on genotype.     

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.     

Littrow-type external-cavity blue laser for holographic data storage

An external-cavity laser with a wavelength of 405 nm and an output of 80 mW has been developed for holographic data storage. The laser has three states: the first is a perfect single mode, whose coherent length is 14 m; the second is a three-mode state with a coherent length of 3 mm; and the third is a six-mode state with a coherent length of 0.3 mm. The first and second states are available for angular-multiplexing recording; all states are available for coaxial multiplexing recording. Due to its short wavelength, the recording density is higher than that of a 532 nm laser.     

Orthogonal sample design scheme for two-dimensional synchronous spectroscopy and its application in probing intermolecular interactions

This paper introduces a new approach to probing intermolecular interactions based on a framework of two-dimensional (2D) synchronous spectroscopy. Mathematical analysis performed on 2D synchronous spectra using variable concentration as an external perturbation shows that the cross-peaks are composed of two parts. The first part reflects intermolecular interactions that manifest in the form of deviation from the Beer-Lambert law. The second part is related simply to the concentration variations of the solutes and is responsible for the generation of interfering cross-peaks not related to the intermolecular interactions in the system. It is the second part that prevents the reliable identification of intermolecular interactions. We propose a way of selecting the concentrations of solutes so that the resultant dynamic concentration vectors of different solutes become orthogonal to one another. Therefore, the contribution of the second part to the cross-peaks can be effectively removed by the dot product of orthogonal vectors. Our new approach has been tested on a simulated chemical system and a real chemical system. The results demonstrate that interfering cross-peaks can be successfully removed from a 2D synchronous spectrum so that the cross-peaks can be used as a reliable tool to characterize or probe intermolecular interactions.     

Thermal degradation of poly(3-hydroxybutyrate) and poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) in nitrogen and oxygen studied by thermogravimetric-Fourier transform infrared spectroscopy

The thermal degradation behavior of poly(3-hydroxybutyrate) (PHB) and poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (P(HB-co-HHx), HHx=12 mol%) has been studied under different environmental conditions by thermogravimetric analysis (TGA) Fourier transform infrared (FT-IR) spectroscopy. It is reported that at higher temperature (>400 degrees C) carbon dioxide and propene are formed from the decomposition product crotonic acid in a nitrogen atmosphere, whereas in an oxygen atmosphere propene oxidizes in a further step to carbon dioxide, carbon monoxide and hydrogen. It was also found that PHB and P(HB-co-HHx) have a similar thermal degradation mechanism. The analysis of the FT-IR-spectroscopic data was performed with 2D and perturbation-correlation moving-window 2D (PCMW2D) correlation spectroscopy.     

Holographic femtosecond laser processing using optimal-rotation-angle method with compensation of spatial frequency response of liquid crystal spatial light modulator

Holographic femtosecond laser processing performs high-speed parallel processing using a computer-generated hologram (CGH) displayed on a liquid crystal spatial light modulator. A critical issue is to precisely control the intensities of the diffraction peaks of the CGH. We propose a method of compensating for the spatial frequency response in the design of CGH using the optimal-rotation-angle method. By applying the proposed method, the uniformity of the diffraction peaks was improved. We demonstrate holographic femtosecond laser processing with two-dimensional and three-dimensional parallelism.     

Therapy of myeloma in vivo using marine phospholipid in combination with Agaricus blazei Murill as an immune respond activator

Mushroom (Agaricus blazei Murill) extract has been reported to possess antitumor effects through immune activation. Here, we investigated the beneficial effects of combining A. blazei extract with marine phospholipids in comparison to A. blazei extract alone on myeloma sp2 tumor suppression when orally administrated. The experimental groups designed for sp2 tumor bearing BALB/c nu/nu mice were drinks of: (1)control; (2)1.0 mg/mL squid phospholipid liposome alone; (3)0.5 mg/mL A. blazei Murill water extract alone; (4)1.0 mg/mL squid phospholipid liposome with 0.5 mg/mL A. blazei Murill water extract in the form of those simple mixture; and (5)1.0 mg/mL squid phospholipid liposome with 0.5 mg/mL A. blazei Murill water extract partially encapsulated. Orally administrated volumes amounted to approximately 5 mL per day per mouse for all groups. A. blazei Murill water extract alone and squid phospholipid alone served groups show moderate tumor suppression with total administrations of approximately 105 mg/mouse for squid phospholipid through out the experimental term. When both A. blazei Murill water extract and squid phospholipid were administrated simultaneously in a simple mixture form, promotional effect on cancer tumor suppression was observed. And when A. blazei Murill water extract was partially encapsulated in the squid phospholipid liposomes with total administrations being 105 mg/mouse for squid phospholipid, effect on cancer tumor suppression was more pronounced. Though there was no statistically significant difference in tumor sizes between the simple mixture form administrated group i.e. group (4) and the partially encapsulated form administrated group i.e. group (5), the tumor vanished mouse was seen in the partially encapsulated form administrated group. Thus it was concluded that combinational administration of the A. blazei Murill water extract and the marine phospholipid may be useful in myeloma sp2 therapy.     

On Implicit Second-Order Ordinary Differential Equations: Completely Integrable and Clairaut Type

In this paper, we give a characterization of implicit second-order ordinary differential equations with smooth complete integrals which we call Clairaut-type equations. Moreover, we consider properties of the Clairaut-type equations and present the duality among special completely integrable equations with respect to Engel–Legendre transformations.      

A unimolecular nanocapsule: Encapsulation property of amphiphilic polymer based on hyperbranched polythreitol

Hyperbranched polythreitol (1) with different molecular weights (M_w,SLS: 1.18 × 10⁴ and 4.79 × 10⁴) was reacted with trityl chloride in DMF to afford a novel amphiphilic polymer (2) consisting of 1 as the hydrophilic core and the trityl groups as the hydrophobic shell. Compound 2 was tested for its ability to act as a unimolecular nanocapsule toward the water-soluble dye, rose bengal (RB). Their encapsulation and release properties were also evaluated by comparison with the degree of substitution (DS) of the trityl groups, i.e., the hydrophobic shell density. The polymers were found to have very good unimolecular nanocapsule characteristics even at extremely low concentrations. The average number of RBs per polymer molecule depended on the hydrophilic core size and the hydrophobic shell density. The increasing DS value led to a decrease in the encapsulated amount due to the decrease in the hydrophilic core space, while the low DS value (less than ca. 20 mol%) led to a destabilization as a unimolecular nanocapsule and a lower encapsulation ability. In particular, 2 with ca. 23% DS value showed an efficient encapsulation. Based on a release test of the RB-loaded unimolecular nanocapsules, the polymers showed a high RB-holding ability in water.     

New π-conjugated polymers containing oxazole in the main chain: Optical and electrochemical properties

Oxazole-containing π-conjugated polymers, an alternating copolymer of 9,9-dioctylfluorene-2,7-diyl and 2,5-bis(p-phenylene)oxazole (P(Flu-DPOxz)) and an alternating copolymer of 2,5-dialkoxy-p-phenylene and 2,5-bis(p-phenylene(oxazole)) (P(ROPh-DPOxz)), have been synthesized by Pd-catalyzed organometallic polycondensation. They were soluble in organic solvents, and had number average molecular weights of 18?600 and 24?700, respectively, in GPC analysis. P(Flu-DPOxz) and P(ROPh-DPOxz) showed UV-vis absorption peaks at about 380 nm in CHCl₃. Powder XRD patterns and UV-vis data of the polymers indicated that the polymer solid was amorphous. The polymers were photoluminescent both in chloroform and in solid. In chloroform, the emission peak appeared at about 420 nm with a high quantum yield of about 70%. The polymers were electrochemically active, and electrochemical data revealed that the oxazole unit behaved as a moderate electron-accepting unit.