Relative reactivity of phenylboronic acid to phenylboronate ion toward L-lactate and mandelate at physiological pH

Kinetic studies were performed on the reactions of phenylboronic acid with L-lactic acid and mandelic acid in acidic aqueous and alkaline solutions in order to specify reactive species in these reactions. It was confirmed that the diprotonated ligand (H₂L: L-lactic acid or mandelic acid) is less reactive than the monoprotonated ligand (HL^?: L-lactate ion or mandelate ion), which made possible direct determination of the rate constants of phenylboronic acid (PhB(OH)₂) and its conjugate base, phenylboronate ion (PhB(OH)₃^?). It was found that PhB(OH)₂ is more reactive than PhB(OH)₃^?. On the basis of kinetic results, it was concluded that the most reactive species are PhB(OH)₂ and HL^? at physiological pH 7.4, so the reaction in the boronic acid-based sensor for L-lactate mainly would occur between these species.     

Estrogen Stimulates Proliferation and Differentiation of Neural Stem/Progenitor Cells through Different Signal Transduction Pathways

Our previous study indicated that both 17β-estradiol (E2), known to be an endogenous estrogen, and bisphenol A (BPA), known to be a xenoestrogen, could positively influence the proliferation or differentiation of neural stem/progenitor cells (NS/PCs). The aim of the present study was to identify the signal transduction pathways for estrogenic activities promoting proliferation and differentiation of NS/PCs via well known nuclear estrogen receptors (ERs) or putative membrane-associated ERs. NS/PCs were cultured from the telencephalon of 15-day-old rat embryos. In order to confirm the involvement of nuclear ERs for estrogenic activities, their specific antagonist, ICI-182,780, was used. The presence of putative membrane-associated ER was functionally examined as to whether E2 can activate rapid intracellular signaling mechanism. In order to confirm the involvement of membrane-associated ERs for estrogenic activities, a cell-impermeable E2, bovine serum albumin-conjugated E2 (E2-BSA) was used. We showed that E2 could rapidly activate extracellular signal-regulated kinases 1/2 (ERK 1/2), which was not inhibited by ICI-182,780. ICI-182,780 abrogated the stimulatory effect of these estrogens (E2 and BPA) on the proliferation of NS/PCs, but not their effect on the differentiation of the NS/PCs into oligodendroglia. Furthermore, E2-BSA mimicked the activity of differentiation from NS/PCs into oligodendroglia, but not the activity of proliferation. Our study suggests that (1) the estrogen induced proliferation of NS/PCs is mediated via nuclear ERs; (2) the oligodendroglial generation from NS/PCs is likely to be stimulated via putative membrane-associated ERs.     

Population structure of Cladophora-borne Escherichia coli in nearshore water of Lake Michigan

We previously reported that the macrophytic green alga Cladophora harbors high densities (up to 10(6) colony-forming units/g dry weight) of the fecal indicator bacteria, Escherichia coli and enterococci, in shoreline waters of Lake Michigan. However, the population structure and genetic relatedness of Cladophora-borne indicator bacteria remain poorly understood. In this study, 835 E. coli isolates were collected from Cladophora tufts (mats) growing on rocks from a breakwater located within the Indiana Dunes National Lakeshore in northwest Indiana. The horizontal fluorophore enhanced rep-PCR (HFERP) DNA fingerprinting technique was used to determine the genetic relatedness of the isolates to each other and to those in a library of E. coli DNA fingerprints. While the E. coli isolates from Cladophora showed a high degree of genetic relatedness (92% similarity), in most cases, however, the isolates were genetically distinct. The Shannon diversity index for the population was very high (5.39). Both spatial and temporal influences contributed to the genetic diversity. There was a strong association of isolate genotypes by location (79% and 80% for lake- and ditch-side samplings, respectively), and isolates collected from 2002 were distinctly different from those obtained in 2003. Cladophora-borne E. coli isolates represented a unique group, which was distinct from other E. coli isolates in the DNA fingerprint library tested. Taken together, these results indicate that E. coli strains associated with Cladophora may be a recurring source of indicator bacteria to the nearshore beach.     

Relationships between Bacteroides 16S rRNA genetic markers and presence of bacterial enteric pathogens and conventional fecal indicators

Occurrence and prevalence of different bacterial enteric pathogens as well as their relationships with conventional (total and fecal coliforms) and alternative fecal indicators (host-specific Bacteroides 16S rRNA genetic markers) were investigated for various water samples taken from different sites with different degrees of fecal contamination. The results showed that a wide range of bacterial pathogens could be detected in both municipal wastewater treatment plant samples and in surface water samples. Logistic regression analysis revealed that total and human-specific Bacteroides 16S rRNA genetic markers showed significant predictive values for the presence of Escheriachia coli O-157, Salmonella, heat-labile enterotoxin (LT) of enterotoxigenic E. coli (ETEC), and heat-stable enterotoxin for human (STh) of ETEC. The probability of occurrence of these pathogenic bacteria became significantly high when the concentrations of human-specific and total Bacteroides 16S rRNA genetic markers exceeded 10(3) and 10(4) copies/100 mL. In contrast, Clostridium perfringens was detected at high frequency regardless of sampling sites and levels of Bacteroides 16S rRNA genetic markers. No genes related to Shigella spp., Staphylococcus aureus and Vibrio cholerae were detected in any samples analyzed in this study. Conventional indicator microorganisms had low levels of correlation with the presence of pathogens as compared with the alternative fecal indicators. These results suggested that real-time PCR-based measurement of alternative Bacteroides 16S rRNA genetic markers was a rapid and sensitive tool to identify host-specific fecal pollution and probably associated bacterial pathogens. However, since one fecal indicator might not represent the relative abundance of all pathogenic bacteria, viruses and protozoa, combined application of alternative indicators with conventional ones could provide more comprehensive pictures of fecal contamination, its source and association with pathogenic microorganisms.     

Noncorrosive separator materials for electric double layer capacitor

The paper evaluates noncorrosive and inexpensive materials, namely polypropylene sheet, fiberglass, and glass wool, as potential separator materials for electric double‐layer capacitor (EDLC) application. Using these materials as separators and the same activated carbon electrodes, properties of two‐electrode capacitors filled with aqueous sulfuric acid (H₂SO₄) were investigated by cyclic voltammetry (CV) as well as galvanostatic and electrochemical impedance spectroscopy (EIS) measurements. Performance comparison of the tested capacitors with an identical capacitor with conventional cellulose separator was also carried out. As a benchmark, the noncorrosive‐separator‐based capacitors demonstrate comparable power and energy densities to those of a cellulose separator, with the highest specific capacitance of 131 F/g and lowest equivalent series resistance of 13 Ω for the glass wool separator. Application of such noncorrosive separators may realize the utilization of high‐concentration aqueous electrolytes, leading to higher rating EDLCs at lower cost compared to organic‐solution‐based capacitors. © 2014 Institute of Electrical Engineers of Japan. Published by John Wiley & Sons, Inc.     

Functional characterization and evolutionary implication of the internal 157-amino-acid sequence of an exoinulinase from Penicillium sp. strain TN-88

An extracellular exoinulinase from the filamentous fungus Penicillium sp. strain TN-88 has a 14-fold higher specific activity of 743 U/mg toward inulin than its equivalent from the Aspergillus niger strain 12 and possesses an internal 157-amino-acid sequence whose corresponding region is absent in the A. niger enzyme. On the basis of sequence alignment, the internal region D' encoding the 157-amino-acid sequence in the Penicillium exoinulinase gene inuD cDNA was inserted into the site between the nucleotides 897 and 898 of the A. niger exoinulinase gene inuE cDNA. The resultant inuE::D' fusion was expressed in the methylotrophic yeast Pichia pastoris. The K(m) value of the secreted hybrid enzyme InuE::D' for inulin hydrolysis was about 1/15 that of the A. niger InuE, whereas its k(cat) value did not differ greatly from that of the InuE. These observations indicate that the Penicillium exoinulinase has evolved by the horizontal transfer and integration of a relevant DNA segment and that the internal sequence D' functions as an additional noncatalytic inulin-affinity region.     

Thermal deformation measurement of the solder joints in electronic packages using electron moiré method

In this paper, electron moiré method is used to measure the thermal deformation of electronic packages. In order to observe the electron moiré fringe, a holographic grid was replicated on the cross section of the BGA type package at 150°C. This grid was fabricated on a glass plate using a moving point holographic system, and was replicated to measured area at high temperature. Under SEM, the holographic grid(specimen grid) and a programmed electron beam scan(master grid) interfere and form electron moiré patterns. The shear strain measurement technique using electron moiré method is described. Using the electron moiré method, the shear strains in the different solder joints were measured and analysed. Some useful results were obtained.