Mode of photocatalytic bactericidal action of powdered semiconductor TiO2 on mutans streptococci.

Powdered semiconductor TiO2 has a photocatalytic bactericidal capacity on some kinds of bacteria, but its mechanism still remains unclear. The mode of its photocatalytic bactericidal action on the mutans group of streptococci was investigated. Powdered TiO2 had a bactericidal capacity on all serotypes of mutans streptococci. Streptococcus sobrinus AHT was mainly used for these experiments. The most effective concentration of TiO2 was about 1 mg ml-1 and, at this concentration, 10(5) colony-forming units of S. sobrinus AHT per millilitre were completely killed within 1 min. In order to search for the mechanism of this effect, a high bacterial cell density (10(9) colony-forming units ml-1) was used in the following studies. "Rapid" leakage of potassium ions from the bacteria occurred parallel to the decrease in cell viability. Protein and RNA were "slowly" released from bacterial cells for a reaction time up to 120 min. The pH of the reaction mixture decreased continuously to 4.5 after 120 min. Co-aggregation of S. sobrinus AHT and powdered TiO2 occurred at high bacterial densities (above 10(8) colony-forming units ml-1). Aggregates gradually decomposed with light irradiation. Transmission electron microscopy of S. sobrinus AHT after photocatalytic action for 60-120 min indicated complete destruction of bacterial cells. From these results, bacterial death appears to be caused by a significant disorder in cell membranes and finally the cell walls were decomposed.     

FTIR detection of protonation/deprotonation of key carboxyl side chains caused by redox change of the Cu(A)-heme a moiety and ligand dissociation from the heme a3-Cu(B) center of bovine heart cytochrome c oxidase

FTIR spectral changes of bovine cytochrome c oxidase (CcO) upon ligand dissociation from heme a(3)() and redox change of the Cu(A)-heme a moiety (Cu(A)Fe(a)()) were investigated. In a photosteady state under CW laser illumination at 590 nm to carbonmonoxy CcO (CcO-CO), the C-O stretching bands due to Fe(a3)()(2+)CO and Cu(B)(1+)CO were identified at 1963 and 2063 cm(-)(1), respectively, for the fully reduced (FR) state [(Cu(A)Fe(a)())(3+)Fe(a3)()(2+)Cu(B)(1+)] and at 1965 and 2061 cm(-)(1) for the mixed valence (MV) state [(Cu(A)Fe(a)())(5+)Fe(a3)()(2+)Cu(B)(1+)] in H(2)O as well as in D(2)O. For the MV state, however, another band due to Cu(B)(1+)CO was found at 2040 cm(-)(1), which was distinct from the alpha/beta conformers in the spectral behaviors, and therefore was assigned to the (Cu(A)Fe(a)())(4+)Fe(a3)()(3+)Cu(B)(1+)CO generated by back electron transfer. The FR-minus-oxidized difference spectrum in the carboxyl stretching region provided two negative bands at 1749 and 1737 cm(-)(1) in H(2)O, which were apparently merged into a single band with a band center at 1741 cm(-)(1) in D(2)O. Comparison of these spectra with those of bacterial enzymes suggests that the 1749 and 1737 cm(-)(1) bands are due to COOH groups of Glu242 and Asp51, respectively. A similar difference spectrum of the carboxyl stretching region was also obtained between (Cu(A)Fe(a)())(3+)Fe(a3)()(2+)Cu(B)(1+)CO and (Cu(A)Fe(a)())(5+)Fe(a3)()(2+)Cu(B)(1+)CO. The results indicate that an oxidation state of the (Cu(A)Fe(a)()) moiety determines the carboxyl stretching spectra. On the other hand, CO-dissociated minus CO-bound difference spectra in the FR state gave rise to a positive and a negative peaks at 1749 and 1741 cm(-)(1), respectively, in H(2)O, but mainly a negative peak at 1735 cm(-)(1) in D(2)O. It was confirmed that the absence of a positive peak is not caused by slow deuteration of protein. The corresponding difference spectrum in the MV state showed a significantly weaker positive peak at 1749 cm(-)(1) and an intense negative peak at 1741 cm(-)(1) (1737 cm(-)(1) in D(2)O). The spectral difference between the FR and MV states is explained satisfactorily by the spectral change induced by the electron back flow upon CO dissociation as described above. Thus, the changes of carboxyl stretching bands induced both by oxidation of (Cu(A)Fe(a)()) and dissociation of CO appear at similar frequencies ( approximately 1749 cm(-)(1)) but are ascribed to different carboxyl side chains.     

Self-assemblies of enzymatically degradable amphiphilic oligopeptides as nonviral gene carrier

Novel biodegradable oligopeptide-type gene carriers composed of cationic residues (KRRRKRKRRRKRKRRC) and oligo leucine segments were developed. The amphiphilic carrier was found to form micelle-like assemblies in aqueous solutions, when the oligo leucine is 12 amino acids length (Pep-L12). NMR, CMC, and GPC analysis revealed their hydrophobic/cationic core/shell morphology. Hydrophobic interaction between leucines is thought to be the major driving force behind formations of assemblies. The transient expression of luciferase introduced to COS-1 cells using Pep-L12 below the CMC is as low as that by the control cationic peptides without leucine residue (Pep-L0), while improved transgene expression was observed in the case of Pep-L12 above CMC. The self-assembly raised the apparent molecular weight and gene transfection ability without loosening their low cytotoxicity. These results indicate that the amphiphilic oligopeptides are very promising materials as highly efficient and less toxic gene carriers.     

Functors on the category of quasi-fibrations

We consider the following questions: when can we extend a continuous endofunctor on Top the category of topological spaces to a fibrewise continuous endofunctor on Top(2) the category of continuous maps? If this is true, does such fibrewise continuous endofunctor preserve fibrations? In this paper, we define Fib the topological category of cell-wise trivial fibre spaces over polyhedra and show that any continuous endofunctor on Top induces a fibrewise continuous endofunctor on Fib preserving the class of quasi-fibrations.     

Determination of vitamin D2 in emulsified nutritional supplements by solid-phase extraction and column-switching high-performance liquid chromatography with UV detection

This paper deals with a method for solid-phase extraction of trace amounts of vitamin D2 (VD2, 19 ng/g) from emulsified nutritional supplements, which contain 50 kinds of compounds, followed by column-switching high-performance liquid chromatography (HPLC) with UV detection at 265 nm. VD2 is present at 1000-20,000,000 times lower concentration than other components. Bond Elut C18 cartridge was chosen as for the emulsified nutritional supplements after comparison with eight other types. A sample solution was applied to the solid-phase extraction cartridge and VD2 was eluted by methanol followed by HPLC. The effects of sample pH, eluent composition and eluate volume on the retention and elution of VD2 on Bond Elut C18 cartridge were examined. The resulting method was simple, rapid (analysis time: approximately 20 min), sensitive (detection limit: approximately 0.1 ng per injection (200 microl) at a signal-to-noise ratio 3:1), and reproducible (relative standard deviation: approximately 6.2%, n=5). The calibration graph for VD2 was linear in the range of 0.1-3 ng per injection (200 microl). Recovery of VD2 was approximately 80% by the standard addition method.     

Synthesis and crystal structure of a Pr5Ni19 superlattice alloy and its hydrogen absorption-desorption property

The intermetallic compound Pr(5)Ni(19), which is not shown in the Pr-Ni binary phase diagram, was synthesized, and the crystal structure was investigated by X-ray diffraction (XRD) and transmission electron microscopy (TEM). Two superlattice reflections with the Sm(5)Co(19)-type structure (002 and 004) and the Pr(5)Co(19)-type structure (003 and 006) were observed in the 2θ region between 2° and 15° in the XRD pattern using Cu Kα radiation. Rietveld refinement provided the goodness-of-fit parameter S = 6.7 for the Pr(5)Co(19)-type (3R) structure model and S = 1.7 for the Sm(5)Co(19)-type (2H) structure model, indicating that the synthesized compound has a Sm(5)Co(19) structure. The refined lattice parameters were a = 0.50010(9) nm and c = 3.2420(4) nm. The high-resolution TEM image also clearly revealed that the crystal structure of Pr(5)Ni(19) is of the Sm(5)Co(19) type, which agrees with the results from Rietveld refinement of the XRD data. The P-C isotherm of Pr(5)Ni(19) in the first absorption was clearly different from that in the first desorption. A single plateau in absorption and three plateaus in desorption were observed. The maximum hydrogen storage capacity of the first cycle reached 1.1 H/M, and that of the second cycle was 0.8 H/M. The 0.3 H/M of hydrogen remained in the metal lattice after the first desorption process.     

Routine high-performance liquid chromatographic determination of ascorbic acid in foods using L-methionine for the pre-analysis sample stabilization

The possibility of use of phosphoric acid (0.2% v/v, pH 2.1) in the mobile phase and co-existing compounds present in foods as the dissolving agent for the pre-analysis sample stabilization were examined for the routine determination of ascorbic acid (AA) in foods by high-performance liquid chromatography (HPLC) with electrochemical detection (ED). The applied potential was set at 400 mV versus an Ag/AgCl reference electrode. It was demonstrated that 0.2% v/v phosphoric acid was the useful mobile phase and l-methionine was the most effective dissolving agent for the pre-run sample stabilization of AA in foods after comparison with other amino acids and water-soluble vitamins. The proposed method was simple, rapid (retention time @ ca. 4 min), sensitive (detection limit: ca. 0.1 ng per injection (5 μl) at a signal-to-noise ratio of 3), highly selective and reproducible (relative standard deviation (R.S.D.); 2.5% (n=7), between-day R.S.D.: 3.7% (5 days)). The calibration graph of AA was linear in the range of 0.1-12.5 ng per injection (5 μl). Recovery of AA was over 90% by the standard addition method. Relationship between structure of compounds and the stability of AA was also examined.     

Splitting off rational parts in homotopy types

It is known algebraically that any abelian group is a direct sum of a divisible group and a reduced group (see Theorem 21.3 of [L. Fuchs, Infinite Abelian Groups, vol. I, Academic Press, New York-London, 1970]). In this paper, conditions to split off rational parts in homotopy types from a given space are studied in terms of a variant of Hurewicz map, say and generalised Gottlieb groups. This yields decomposition theorems on rational homotopy types of Hopf spaces, T-spaces and Gottlieb spaces, which has been known in various situations, especially for spaces with finiteness conditions.     

A new catalytic electrode for the amperometric determination of hydrogen peroxide

The electrode is based on the decomposition of H₂O₂ by an inorganic polymer catalyst. The steady -state decomposition currents of H₂0₂ obtained with the catalyst electrode are independent of ionic strength (0.1–1.5) and pH (2.5–10.5). A linear relationship is obtained from 0.02 to 2 mM H₂0₂ in 0.1 M KCl at 25°C.