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.     

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     

A Model of the Relationship Between Psychosocial Variables and Diurnal Cortisol Rhythm Under Chronic Stress by Using Structural Equations

By using structural equations, we investigated the effect of chronic stress on salivary cortisol rhythm and proposed a causal model of chronic stress by using psychosocial and physiological data. First, 111 healthy workers (48 males, 63 females) completed questionnaires on chronic stress and lifestyle habits. Then, they provided saliva samples and answered questionnaires that were prepared to assess their psychological states 5 times (on waking up and at 10:00, 11:40, 14:00, and 16:00) on workdays. Structural equation modeling (SEM) revealed that chronic stress and longer commuting time resulted in sleep irregularities and this disrupted the cortisol circadian rhythm. This suggests that chronic stress disrupts the cortisol circadian rhythm even in healthy individuals, and sleep regularity mediates the effect of chronic stress on the cortisol rhythm.

     

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.     

Air vent in water chamber and surface coating on liner slides concerning auxiliary cooling system for the high temperature engineering test reactor

Experimental study was made to confirm the validity of new designs of the auxiliary cooling system for the high temperature engineering test reactor (HTTR). First, it is necessary to vent residence air in outlet side of water chamber of the auxiliary heat exchanger for the HTTR. Accordingly, we have proposed to mount a proper bend duct in the outlet side of the water chamber. Air vent is done by difference between pressures at both ends of the bend duct caused by the forced water circulation using the water pumps. From flow tests, it was confirmed that it is capable of venting the air through the bend duct by circulating the water in maximum capacity of the water pumps. Second, it is essential to prevent seizure and excessive wear of the liner slides of the auxiliary concentric hot gas duct for the HTTR at a service temperature of 950°C. Therefore, we have put forward to coat titanium nitride (TiN) on the surface of the liner slides made of nickel-based superalloy Hastelloy XR using the thermochemical vapor deposition method. As a result of seizure and wear tests, it was confirmed that the TiN coating film of 3 μm on the surface of Hastelloy XR is sufficient.     

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.