Real-Time Duplex Applications of Loop-Mediated AMPlification (LAMP) by Assimilating Probes

Isothermal nucleic-acid amplification methods such as Loop-Mediated isothermal AMPlification (LAMP) are increasingly appealing alternatives to PCR for use in portable diagnostic system due to the low cost, weight, and power requirements of the instrumentation. As such, interest in developing new probes and other functionality based on the LAMP reaction has been intense. Here, we report on the development of duplexed LAMP assays for pathogen detection using spectrally unique Assimilating Probes. As proof of principle, we used a reaction for Salmonella enterica as a model coupled with a reaction for λ-phage DNA as an internal control, as well as a duplexed assay to sub-type specific quarantine strains of the bacterial wilt pathogen Ralstonia solanacearum. Detection limits for bacterial DNA analyzed in individual reactions was less than 100 genomic equivalents in all cases, and increased by one to two orders of magnitude when reactions were coupled in duplexed formats. Even so, due to the more robust activity of newly available strand-displacing polymerases, the duplexed assays reported here were more powerful than analogous individual reactions reported only a few years ago, and represent a significant advance for incorporation of internal controls to validate assay results in the field.     

Matrix influence on the piezoelectric properties of piezoelectric ceramic/polymer composite exhibiting particle alignment

This study was addressed to the influence of an electric field strength applied at fabrication process and matrix properties, such as the dielectric constant and the Young's modulus, on “pseudo‐1‐3 piezoelectric ceramic/polymer composite” in order to further enhance the piezoelectricity of that. The pseudo‐1‐3 piezoelectric ceramic/polymer composite consists of linearly ordered piezoelectric ceramic particles in polymer material. Silicone gel, silicone rubber, urethane rubber, and poly‐methyl‐methacrylate, which exhibit different dielectric constants and Young's modulus, were used as matrices to evaluate the matrix influence. The piezoelectricity of the pseudo‐1‐3 piezoelectric ceramic/polymer composite was evaluated using the piezoelectric strain constant d₃₃. The d₃₃ is one of the indices of the piezoelectric properties for piezoelectric materials. As a result, it was confirmed that d₃₃ of the pseudo‐1‐3 piezoelectric ceramic/polymer composite increased with the increase of the electric filed strength applied at fabrication process, though, it reached a constant value at a certain strength value. Further it was confirmed that dielectric constant of the matrix had a small influence on d₃₃ of the pseudo‐1‐3 piezoelectric ceramic/polymer composite, however, in case of matrix of lower Young's modulus, d₃₃ was increase. © 2014 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2015 , 132, 41817.     

An optical instrumentation using a sensor with a sensordummyagainst noises on sensor and transmission line

This short paper discusses the method of effectively canceling equal status normal mode noise not only on a sensor line but also on a transmission line of an optical instrument using a sensor with a sensordummy resistance     

Unperturbed dimension of poly(4-acetoxystyrene) measured in good and theta solvents

In order to obtain the additional data concerning the unperturbed dimension of poly-4-substituted styrene, light scattering measurements are performed on the twenty-two fractions with molecular weight of (0.91–352)·10⁴ of poly(4-acetoxystyrene) in dioxan at 25°C, from which the molecular weight obtained was found easily to be evaluated with the gel permeation chromatography using THF. Phase separation experiments for this polymer indicate that the theta state is attained in isopropyl acetate at 19.7°C and butyl acetate at 26.8°C. By making viscosity measurements at that state, the value of K_Θ is directly evaluated as 5.4·10^?4 dl g^?1. The limiting viscosity number is also obtained in good solvents, THF and dioxan, at 25°C and constants of the Mark-Houwink-Sakurada equation in each solvent are determined. Further, approximately the same K_Θ as above is obtained from these data with the Stockmayer-Fixman plot. The calculated value of steric factor, 2.37, on this polymer may be plausible, compared with those of polyvinylaromatic derivatives.     

Volumetric properties of mixtures of 1,4-dioxane and water at high pressures

Densities of aqueous of 1,4-dioxane have been measured at temperatures from 298 to 348 K and at pressures up to 40 MPa by a vibrating-tube method. Molar volumes obtained with an estimated uncertainty of ±0.2°,% are correlated with pressure by the Tait equation within the experimental uncertainty. Pressure and composition dependences of the excess molar volume, partial molar volume, and isothermal compressibility are determined and they are compared with those of other aqueous solutions.     

The anti-tumor arotinoid Ro 40-8757 protects bone marrow from the toxic effects of 5-fluorouracil

We have previously shown that strychnine mimics the cytoprotective properties of glycine in renal proximal tubule (RPT) suspensions exposed to antimycin A (AA). The aims of this study were to determine whether the cytoprotective properties of strychnine applied to various types of nephrotoxicants and to examine the temporal aspects of the cytoprotection of glycine and strychnine. Tubular release of LDH activity was used as a marker of cell death. Glycine (2 mM) or strychnine (1 mM) added 5 min prior to the toxicant decreased LDH release in rabbit RPT suspensions exposed to 25 microM tetrafluoroethyl-L-cysteine (TFEC), 10 microM HgCl2, 0.5 mM t-butyl hydroperoxide (TBHP), or 0.2 mM bromohydroquinone (BHQ) for 4 hr, or 2 mM sodium cyanide (NaCN) for 2 hr. The relative rank order of effectiveness of glycine and strychnine was NaCN = TFEC > BHQ > DCVC > TBHP > HgCl2. The temporal aspects of strychnine and glycine protection were examined by exposing RPT to either AA or TFEC for 1 or 3 hr, respectively, and then adding either 1 mM glycine or 1 mM strychnine. Glycine and strychnine decreased LDH release in AA-treated RPT at 1.25 and 2 hr and TFEC-treated RPT at 4 hr. In addition, when RPT exposed to AA or TFEC and treated with strychnine or glycine were washed at either 1 or 4 hr, protection was eliminated at later time points. When glycine was added to RPT treated with either PCBC, TFEC, or DCVC 5 min prior to or 30, 60, 120, and 180 min following toxicant addition, LDH release was reduced at all time points. These results demonstrate that strychnine and glycine protect RPT from a variety of diverse nephrotoxicants, strychnine and glycine do not need to be present at the time of toxic insult, strychnine and glycine cytoprotection is reversible, and strychnine and glycine act in the late phase of necrotic cell injury.     

Regenerating-mode low-speed operation of sensorless induction motor drive with adaptive observer

Speed estimation methods for sensorless induction motor drives can be divided into two groups. One is based on information about fundamental components of stator currents and voltages. The other is based on high-frequency components injection in stator voltages or currents. The latter may operate stably under zero frequency, which occurs in regenerating mode at low speeds. It, however, causes loss increasing and torque ripple. The former is difficult to operate under zero frequency. The authors propose a stable method categorized in the former group even in regenerating mode at low speeds. The proposed method is based on an adaptive flux observer. First, a new observer gain design is shown. Next, a method avoiding zero-frequency operation with controlling a rotor flux level is proposed.     

Diffusion-controlled size-selective chloride ion sensing in the presence of bromide ion using a thin, nanoporous, plasma-polymerized coating on an Ag/AgCl electrode

We have developed a diffusion-controlled size-selective method for sensing chloride ion in the presence of bromide ion, based on a thin, nanoporous, plasma-polymerized coating of hexamethyldisiloxane on an Ag/AgCl electrode. Sub-nanometer-sized pores responsible for a highly cross-linked polymer network in the plasma-polymerized coating allowed diffusion-controlled permeation of chloride ion while blocking the larger bromide ion. An electrode coating of thickness greater than 70 nm enabled chloride detection in the concentration range 1-10 mM in the presence of 0.63 mM bromide ion. Advantages of this approach are: (1) simple design compared with ionophore-based strategies and (2) compatibility with microfabrication and mass production processes.     

Extensions of polyacrylic acid ammonium salts in the adsorption layer to fluidize alumina slurries

An average shell volume occupied in the adsorption layer on alumina by a polyacrylic acid ammonium salts molecule (PAA) defined as the average area occupied by an adsorbed PAA on alumina multiplied by the average thickness of the water layer at the limit of thickening, has been calculated from the adsorbed amount of PAA and from the flow points of alumina in the presence of PAA of different molecular weights. A steric effect of the PAA dispersant on the dispersion of alumina resulted in a flow with no yield stress. This was due to the change of the extension of PAA in the adsorption shell from a sphere to an ellipsoid with a long axis which exceeded the effective distance that the van der Waals attraction force reaches at a molecular weight for the PAA of between 10 000–20 000.