Genetic analysis of double-strand break repair in Escherichia coli

We had reported that a double-strand gap (ca. 300 bp long) in a duplex DNA is repaired through gene conversion copying a homologous duplex in a recB21 recC22 sbcA23 strain of Escherichia coli, as predicted on the basis of the double-strand break repair models. We have now examined various mutants for this repair capacity. (i) The recE159 mutation abolishes the reaction in the recB21C22 sbcA23 background. This result is consistent with the hypothesis that exonuclease VIII exposes a 3'-ended single strand from a double-strand break. (ii) Two recA alleles, including a complete deletion, fail to block the repair in this recBC sbcA background. (iii) Mutations in two more SOS-inducible genes, recN and recQ, do not decrease the repair. In addition, a lexA (Ind-) mutation, which blocks SOS induction, does not block the reaction. (iv) The recJ, recF, recO, and recR gene functions are nonessential in this background. (v) The RecBCD enzyme does not abolish the gap repair. We then examined genetic backgrounds other than recBC sbcA, in which the RecE pathway is not active. We failed to detect the double-strand gap repair in a rec+, a recA1, or a recB21 C22 strain, nor did we find the gap repair activity in a recD mutant or in a recB21 C22 sbcB15 sbcC201 mutant. We also failed to detect conservative repair of a simple double-strand break, which was made by restriction cleavage of an inserted linker oligonucleotide, in these backgrounds. We conclude that the RecBCD, RecBCD-, and RecF pathways cannot promote conservative double-strand break repair as the RecE and lambda Red pathways can.     

Analysis of XANES for identification of highly dispersed transition metal oxides on supports

XANES of vanadium and niobium oxide on silica or alumina have been analyzed quantitatively by a deconvolution technique. Based on the results for reference compounds, local structures of supported vanadium and niobium species were identified. The composition was estimated from difference spectra for the samples which consisted of two kinds of species.     

Wasserabsorptionseigenschaften natürlicher polyionenkomplexe

As the cationic component of natural polymer polyionic complexes, polysaccharide chitosan or polypeptide polylysine, and as the anionic component, polysaccharide carboxymethyl cellulose (=CMC) or polypeptide poly(glutamic acid) (PGA) were used. The degree of chitosan/CMC or polylysine/PGA complex film swelling was minimum, when the amounts of the cationic and anionic groups in these polymers were balanced at 1 : 1 ratio. On the other hand, the degree of chitosan/PGA film swelling was minimum at the ratio with excess of PGA which is a linear flexible chain. Comparing the enthalpies estimated by DSC analysis, it was suggested that the larger quantity of water in the form of nonfreezable bound water was absorbed in the complex films with polysaccharide which is a chain of ring structure. Further, the compressive moduli of swollen complex films were high, when the complex consisted of polysaccharide components.     

Preparation of Copper-Ruby Glasses by Sputtering: The Effect of Atmosphere on the Growth of Copper Particles

Copper-ruby glasses were prepared by the sputtering method and the effect of reducing treatment was examined. The reducing was carried out either during sputtering or heat treatment. Optical absorption was related to the growth of copper particles. The results are summarized as follows: (i) the volume fraction of copper particles in the glasses which are heat-treated in air increases with the addition of hydrogen in the sputtering gas. (ii) The copper particles do not grow larger than 8 nm by heat treatment in reducing atmosphere, whereas they grow as large as 12 nm in air. (iii) Reducing during heat treatment gives a pronounced effect only when sputtering was not carried out under reducing conditions. The origin of these phenomena was also discussed, considering the mechanism of nucleation and growth of copper particles.     

Planar AlGaAs/GaAs heterojunction bipolar transistors fabricatedusing selective W-CVD

The authors describe a planar process for the AlGaAs/GaAs HBTs in which collector vias are buried selectively, even to the base layers, with chemical vapor deposited tungsten (CVD-W) films. By using WF₆ /SiH₄ chemistry, W could be deposited on Pt films, which were overlapped 50 nm thick on the AuGe-based collector electrodes, without depositing W on the surrounding SiO₂ layers. Current gains of planar HBTs with 3.5-μm×3.5-μm emitters were up to 150, for a collector current density of about 2.5×10⁴ A/cm²     

Influence of methylprednisolone on the intracellular antimicrobial activity of erythromycin and clindamycin against Legionella pneumophila

We have investigated the effect of methylprednisolone on the intracellular activity of erythromycin and clindamycin in vitro. An assay system was developed for the determination of intracellular activity of antibiotics against Legionella pneumophila using guinea pig resident alveolar macrophages. Erythromycin at a concentration of 0.625 mg/L (5 x MIC) and clindamycin at a concentration of 8 mg/L (MIC) inhibited the growth of a single strain of L. pneumophila in macrophages, whilst ceftizoxime at a concentration of 0.625 mg/L (5 x MIC) did not. Methylprednisolone at therapeutic concentrations did not affect the intracellular antibacterial activity of either erythromycin or clindamycin against L. pneumophila. We found no direct effect of methylprednisolone on the intracellular antibacterial activity of either erythromycin or clindamycin.     

Simplified quantitative assay system for measuring activities of drugs against intracellular Legionella pneumophila

We developed a new simple assay for the quantitation of the activities of drugs against intracellular Legionella pneumophila. The cells of a murine macrophage-like cell line (J774.1 cells) allowed the intracellular growth and replication of the bacteria, which ultimately resulted in cell death. The infected J774.1 cell monolayers in 96-well microplates were first treated with antibiotics and were further cultured for 72 h. The number of viable J774.1 cells in each well was quantified by a colorimetric assay with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and an enzyme-linked immunosorbent assay reader. The number of growing bacteria in each well was also determined by counting the numbers of CFU on buffered charcoal yeast extract-alpha agar plates. Viable J774.1 cell counts, determined by the colorimetric assay, were inversely proportional to the number of intracellular replicating bacteria. The minimum extracellular concentrations (MIECs) of 24 antibiotics causing inhibition of intracellular growth of L. pneumophila were determined by the colorimetric assay system. The MIECs of beta-lactams and aminoglycosides were markedly higher than the MICs in buffered yeast extract-alpha broth. The MIECs of macrolides, fluoroquinolones, rifampin, and minocycline were similar to the respective MICs. According to their intracellular activities, clarithromycin and sparfloxacin were the most potent among the macrolides or fluoroquinolones tested in this study. Our results indicated that the MTT assay system allows comparative and quantitative evaluations of the intracellular activities of antibiotics and efficient processing of a large number of samples.     

A mesenteric liposarcoma with production of granulocyte colony-stimulating factor

A 77-year-old female was admitted to our hospital because of pyrexia and a right retroperitoneal mass. Leukocytosis and other inflammatory findings were noted. Bone-marrow aspiration revealed hypercellularity with no malignant cells. An additional mass was detected sonographically in the pelvis. The serum concentration of granulocyte colony-stimulating factor (G-CSF) was highly elevated (299 pg/ml). The tumors were removed at laparotomy, and the pelvic mass was found to arise from the ileocecal mesentery. Postoperatively, white blood cell count and serum G-CSF concentrations decreased to normal levels. The mesenteric tumor showed weakly positive immunostaining for human G-CSF, and Northern and polymerase chain reaction (PCR) analyses detected CSF and its mRNA in the mesenteric tumor.     

Preparation of zirconia-toughened bioactive glass-ceramics

Bioactive glass-ceramics toughened by tetragonal zirconia polycrystal (TZP) were prepared by hot-pressing mixed powders of the MgO-CaO-P₂O₅-SiO₂ glass and TZP containing 20 to 80% alumina. The bending strength and the fracture toughness of the composite materials were improved compared with those of the material without TZP. These composites showed high bending strengths (400 to 500MPa) and high fracture toughness ( 2.8MPa m^1/2). The existence of a crack deflection mechanism was observed by scanning electron microscopy. After soaking in simulated physiological solution at 100 °C, no phase transformation from tetragonal to monoclinic of TZP in the composites and no degradation in bending strength occurred.