Evaluation of the impact of bioaugmentation and biostimulation by in situ hybridization and microelectrode

Three rotating disk biofilm reactors were operated to evaluate whether bioaugmentation and biostimulation can be used to improve the start-up of microbial nitrification. The first reactor was bioaugmented during start-up period with an enrichment culture of nitrifying bacteria, the second reactor received a synthetic medium containing NH(4)(+) and NO(2)(-) to facilitate concomitant proliferation of ammonia- and nitrite-oxidizing bacteria, and the third reactor was used as a control. To evaluate the effectiveness of bioaugmentation and biostimulation approaches, time-dependent developments of nitrifying bacterial community and in situ nitrifying activity in biofilms were monitored by fluorescence in situ hybridization (FISH) technique and microelectrode measurements of NH(4)(+), NO(2)(-), NO(3)(-), and O(2). In situ hybridization results revealed that addition of the enrichment culture of nitrifying bacteria significantly facilitated development of dense nitrifying bacterial populations in the biofilm shortly after, which led to a rapid start-up and enhancement of in situ nitrification activity. The inoculated bacteria could proliferate and/or survive in the biofilm. In addition, the addition of nitrifying bacteria increased the abundance of nitrifying bacteria in the surface of the biofilm, resulting in the higher nitrification rate. On the other hand, the addition of 2.1mM NO(2)(-) did not stimulate the growth of nitrite-oxidizing bacteria and did inhibit the proliferation of ammonia-oxidizing bacteria instead. Thus, the start-up of NO(2)(-) oxidation was unchanged, and the start-up of NH(4)(+) oxidation was delayed. In all the three biofilm reactors, data sets of time series analyses on population dynamics of nitrifying bacteria determined by FISH, in situ nitrifying activities determined by microelectrode measurements, and the reactor performances revealed an approximate agreement between the appearance of nitrifying bacteria and the initiation of nitrification activity, suggesting that the combination of these techniques was a very powerful monitoring tool to evaluate the effectiveness of bioaugmentation and biostimulation strategies.     

Determination of potent antisense oligonucleotides in vitro by semiempirical rules

The selection of effective antisense target sites on a given mRNA molecule is a major problem in the detection of target mRNA in oligonucleotide arrays. In general, antisense oligodeoxynucleotides (asODNs) of about 10-20 nucleotides (nt) in length are used. However, the demand for predicting the sequence of potent asODNs much longer than those mentioned above has been increasing. Here, we prepared 40-nt asODNs directed against fluorescence-labeled green fluorescent protein (GFP) mRNA and quantified their hybridization efficiencies by fluorescence microscopy. We found that the hybridization efficiency depended on the TC content or the minimum free energy of the asODNs. On the basis of these findings, a semiempirical parameter called accessibility score was introduced to predict the potency of asODNs. The results of this study aided in the development of an effective two-step procedure for determining mRNA accessibility, namely, the computer-aided selection of asODN binding sites using an accessibility score followed by an experimental procedure for measuring the hybridization efficiencies between the selected asODNs and the target mRNA by fluorescence microscopy.     

The present status of dental titanium casting

Experimentation in all aspects of titanium casting at universities and industries throughout the world for the last 20 years has made titanium and titanium-alloy casting nearly feasible for fabricating sound cast dental prostheses, including crowns, inlays, and partial and complete dentures. Titanium casting in dentistry has now almost reached the stage where it can seriously be considered as a new method to compete with dental casting using conventional noble and base-metal alloys. More than anything else, the strength of titanium’s appeal lies in its excellent biocompatibility, coupled with its comparatively low price and abundant supply. Research efforts to overcome some problems associated with this method, including studies on the development of new titanium alloys suitable for dental use, will continue at many research sites internationally. Toru Okabe earned his Ph.D. in metallurgical and materials engineering at the University of Florida in 1969. He is currently professor and chair of biomaterials science at Baylor College of Dentistry. Dr. Okabe is a member of TMS. Chikahiro Ohkubo earned his Ph.D. in removable prosthodontics and his D.M.D. in dentistry at Tsurumi University in 1989 and 1985, respectively. He is currently an instructor at the School of Dental Medicine at Tsurumi University in Yokohama, Japan. Ikuya Watanabe earned his Ph.D. in dental science at Nagasaki University in 1995. He is currently working at Nagasaki University, Japan. Osamu Okuno earned his Ph.D. in engineering at Waseda University in 1976. He is currently a professor and chair in the Department of Dental Materials Science, School of Dentistry, Tohoku University, Japan. Yukyo Takada earned his Ph.D. in engineering at Waseda University in 1991. He is currently assistant professor in the Department of Dental Materials Science, School of Dentistry, Tohoku University, Japan.     

Gene duplication and multiplicity of collagenases in Clostridium histolyticum

Clostridium histolyticum collagenase contains a number of different active components. Previously we have shown that colH encodes a 116-kDa collagenase (ColH) and a 98-kDa gelatinase. We purified a different 116-kDa collagenase (ColG) from the culture supernatant and sequenced its gene (colG). We also identified four other gelatinases (105, 82, 78, and 67 kDa) and determined their N-terminal amino acid sequences, all of which coincided with that of either ColG or ColH. Hybridization experiments showed that each gene is present in a single copy and each gene is transcribed into a single mRNA. These results suggest that all the gelatinases are produced from the respective full-length collagenase by the proteolytic removal of C-terminal fragments. The substrate specificities of the enzymes suggest that colG and colH encode class I and class II enzymes, respectively. Analysis of their DNA locations by pulsed-field gel electrophoresis and nucleotide sequencing of their surrounding regions revealed that the two genes are located in different sites on the chromosome. C. histolyticum colG is more similar to C. perfringens colA than to colH in terms of domain structure. Both colG and colA have a homologous gene, mscL, at their 3' ends. These results suggest that gene duplication and segment duplication have occurred in an ancestor cell common to C. histolyticum and C. perfringens and that further divergence of the parent gene produced colG and colA.     

System Cu-Rh-O: Phase diagram and thermodynamic properties of ternary oxides CuRhO2 and CuRh2O4

An isothermal section of the phase diagram for the system Cu-Rh-O at 1273 K has been established by equilibration of samples representing eighteen different compositions, and phase identification after quenching by optical and scanning electron microscopy (SEM), X-ray diffraction (XRD), and energy dispersive analysis of X-rays (EDX). In addition to the binary oxides Cu₂O, CuO, and Rh₂O₃, two ternary oxides CuRhO₂ and CuRh₂O₄ were identified. Both the ternary oxides were in equilibrium with metallic Rh. There was no evidence of the oxide Cu₂Rh₂O₅ reported in the literature. Solid alloys were found to be in equilibrium with Cu₂O. Based on the phase relations, two solid-state cells were designed to measure the Gibbs energies of formation of the two ternary oxides. Yttria-stabilized zirconia was used as the solid electrolyte, and an equimolar mixture of Rh+Rh₂O₃ as the reference electrode. The reference electrode was selected to generate a small electromotive force (emf), and thus minimize polarization of the three-phase electrode. When the driving force for oxygen transport through the solid electrolyte is small, electrochemical flux of oxygen from the high oxygen potential electrode to the low potential electrode is negligible. The measurements were conducted in the temperature range from 900 to 1300 K. The thermodynamic data can be represented by the following equations: {fx741-1} where Δ_f(ox) G ^o is the standard Gibbs energy of formation of the interoxide compounds from their component binary oxides. Based on the thermodynamic information, chemical potential diagrams for the system Cu-Rh-O were developed.     

Image analysis of morphological change during arachidonic acid production by Mortierella alpina 1S-4

The changes in mycelial morphology during arachidonic acid (AA) production by Mortierella alpina 1S-4 were investigated using an image analysis system. Cultivation was performed in a 10-kl fermentor, and the culture broth was separated into two fractions by sieving (0.5 mm aperture size): the filament fraction (F-fraction, <0.5 mm), and the pellet fraction (P-fraction, >0.5 mm). The effect of the mycelial morphology in each fraction on AA production was analyzed. As a result, a product distribution in the culture broth wherein the AA content in the mycelia of the P-fraction was observed to be higher than that in the mycelia of the F-fraction throughout the cultivation. Morphological analysis of the P-fraction revealed that the hairy pellets became smooth because the mycelia on the pellet surface were shaved off; some pellets were broken and reduced in size. The shaved-off mycelia from the hairy pellets surface moved into the F-fraction and aggregated there. From the above findings, it was likely that the low AA content in the F-fraction was due to mycelial damage during the cultivation. In addition, the morphology of the hairy pellets was found to contribute to an increase in the viscosity of culture broth.     

System Nd-Pd-O: Phase diagram and thermodynamic properties of oxides using a solid-state cell with advanced features

An isothermal section of the phase diagram for the system Nd-Pd-O at 1350 K has been established by equilibration of samples representing 13 different compositions and phase identification after quenching by optical and scanning electron microscopy, x-ray diffraction, and energy dispersive analysis of x-rays. The binary oxides PdO and NdO were not stable at 1350 K. Two ternary oxides Nd₄PdO₇ and Nd₂Pd₂O₅ were identified. Solid and liquid alloys, as well as the intermetallics NdPd₃ and NdPd₅, were found to be in equilibrium with Nd₂O₃. Based on the phase relations, three solidstate cells were designed to measure the Gibbs energies of formation of PdO and the two ternary oxides. An advanced version of the solid-state cell incorporating a buffer electrode was used for high-temperature thermodynamic measurements. The function of the buffer electrode, placed between reference and working electrodes, was to absorb the electrochemical flux of the mobile species through the solid electrolyte caused by trace electronic conductivity. The buffer electrode prevented polarization of the measuring electrode and ensured accurate data. Yttria-stabilized zirconia was used as the solid electrolyte and pure oxygen gas at a pressure of 0.1 MP a as the reference electrode. Electromotive force measurements, conducted from 950 to 1425 K, indicated the presence of a third ternary oxide Nd₂PdO₄, stable below 1135 (±10) K. Additional cells were designed to study this compound. The standard Gibbs energy of formation of PdO (†_f G ⁰) was measured from 775 to 1125 Kusing two separate cell designs against the primary reference standard for oxygen chemical potential. Based on the thermodynamic information, chemical potential diagrams for the system Nd-Pd-O were also developed.     

Fatigue Life of Cast Titanium Alloys Under Simulated Denture Framework Displacements

The objective of the study was to evaluate the hypothesis that the mechanical properties and fatigue behavior of removable partial dentures (RPD) made from cast titanium alloys can be improved by alloying with low-cost, low-melting elements such as Cu, Al, and Fe using commercially pure Ti (CP-Ti) and Ti-6Al-4V as controls. RPD specimens in the form of rest-shaped, clasp, rectangular-shaped specimens and round-bar tensile specimens were cast using an experimental Ti-5Al-5Cu alloy, Ti-5Al-1Fe, and Ti-1Fe in an Al₂O₃-based investment with a centrifugal-casting machine. The mechanical properties of the alloys were determined by performing tensile tests under a controlled displacement rate. The fatigue life of the RPD specimens was tested by the three-point bending in an MTS testing machine under a cyclic displacement of 0.5 mm. Fatigue tests were performed at 10 Hz at ambient temperature until the specimens failed into two pieces. The tensile data were statistically analyzed using one-way ANOVA (α = 0.05) and the fatigue life data were analyzed using the Kaplan-Meier survival analysis (α = 0.05). The experimental Ti-5Al-5Cu alloy showed a significantly higher average fatigue life than that of either CP-Ti or Ti-5Al-1Fe alloy (p < 0.05). SEM fractography showed that the fatigue cracks initiated from surface grains, surface pores, or hard particles in surface grains instead of the internal casting pores. Among the alloys tested, the Ti-5Al-5Cu alloy exhibited favorable results in fabricating dental appliances with an excellent fatigue behavior compared with other commercial alloys.     

A diagnostic method based on RVM (return voltage measurement) for condenser bushings with oil–paper insulation systems

The bushing of a transformer is one part of the transformer system, but is as vital as the transformer itself because it forms a part of the main circuit. Deterioration and abnormal condition of the bushing possibly cause its destruction, shortage of the main circuit to the Earth, and fire in the systems, so that diagnosis of the bushings is thus very important. At present, mainly visual inspection from outside of the bushing is carried out. Inspection of the inside condition of the bushing by dissolved gas analysis of the inside oil is done only as sampling for selected bushings because it is difficult due to the sealed construction of the bushings. Therefore, a simple diagnostic method for bushings has been expected for a long time. In Eastern Europe, the RVM (Return Voltage Measurement) method based on scanning the polarization spectra was applied to the oil–paper insulation systems inside the transformer as a diagnostic method. In this paper we investigate theoretically the principle of the RVM method applied to bushings with the same oil–paper insulation systems as the transformer by using the equivalent circuit, and the possibility of using this novel diagnostic method for condenser‐type bushings is demonstrated by a model experiment. © 1999 Scripta Technica, Electr Eng Jpn, 130(1): 21–29, 2000     

Synthesis of 2H‐Thiopyrans by Rhodium(II) Acetate‐Catalyzed Reaction of 4‐Amino‐2,5‐dihydro‐3‐thiophenecarbonitriles with α‐Diazocarbonyl Compounds. Part 1

4‐Amino‐2,5‐dihydro‐3‐thiophenecarbonitriles 1 reacted with dimethyl diazomalonate in the presence of rhodium(II) acetate to give regioselectively 4‐cyano‐2H‐thio‐pyrans 2 (C ₂— S insertion), and 5‐cyano‐2H‐thiopyrans (C ₅— S insertion) were not isolated. Similar insertion was also observed in the reaction of 1 with methyl diazoacetoacetate and ethyl diazobenzoylacetate. The starting compounds 1 were synthesized by the reaction of tetrahydro‐4‐oxo‐3‐thiophene‐carbonitrile with morpholine, piperidine, and pyrrolidine in the presence of formic acid in ethanol.