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.     

Two patients with polymyositis or dermatomyositis complicated with massive pleural effusion

Two patients with polymyositis (PM) or dermatomyositis (DM) complicated with massive pleural effusion are reported here. Both patients presented a high-grade fever, pleural effusion prominent on the right, and good response to steroid therapy. In a 50-year-old woman with PM, combined process of pleural inflammation, cardiomyopathy and coexisting hypothyroidism were considered to be responsible for the accumulation of the massive pleural effusion. However, in a 34-year-old man with DM, pleural inflammation associated with interstitial pneumonia or pleural microvasculopathy in DM was considered to be responsible for the accumulation of the massive pleural effusion.     

Evidence that angiotensin II, endothelins and nitric oxide regulate mitogen-activated protein kinase activity in rat aorta

We measured the activity of mitogen-activated protein (MAP) kinases, enzymes believed to be involved in the pathway for cell proliferation, in rat aortic strips with or without endothelium, and examined effects of angiotensin receptor antagonists, endothelin receptor antagonists and nitric oxide (NO)-related agents. Endothelium removal produced an activation of MAP kinase activity in the strips, whereas the enzyme activity was not affected in the adventitia. The MAP kinase activation was inhibited by either the angiotensin AT1 receptor antagonist losartan or the endothelin ETA receptor antagonist BQ 123. The combination of both antagonists caused an additive inhibition. The angiotensin AT2 receptor antagonist PD 123,319 and the endothelin ETB receptor antagonist BQ 788 did not affect the MAP kinase activation. The NO synthase inhibitor NG-nitro-L-arginine methyl ester (L-NAME) caused an activation of MAP kinase in the endothelium-intact aorta and the MAP kinase activation was inhibited by losartan or BQ123. The NO releaser nitroprusside inhibited the MAP kinase activation induced by endothelium removal or angiotensin II. These results suggest that even in isolated arteries, NO of endothelial origin tonically exert MAP kinase-inhibiting effects and endogenous angiotensin II and endothelins in the media are tonically released to cause MAP kinase-stimulating effects in medial smooth muscle.     

Decreased membrane fluidity and unsaturated fatty acids in Niemann-Pick disease type C fibroblasts

We have investigated transformation with heterologous DNA as a method for insertional mutagenesis of Aspergillus fumigatus. Two methods, polyethylene glycol-mediated transformation of protoplasts and electroporation of germinating spores, were used to establish conditions leading to single-copy integration of transforming DNA at different genomic sites. We have assessed the effect of restriction enzyme-mediated integration (REMI) for both methods. Non-REMI protoplast transformation led to integration of multiple copies of transforming DNA in the majority of transformants. Results of REMI with protoplast transformation varied depending on the enzyme used. Low concentrations of several restriction enzymes stimulated transformation, but of ten enzymes investigated only REMI with XhoI and KpnI resulted in single-copy integration of transforming DNA for the majority of transformants. For protoplast transformation with XhoI- or KpnI-based REMI, 50% and 76% of insertions, respectively, were due to integrations at a genomic enzyme site corresponding to the enzyme used for REMI. Electroporation of spores without addition of restriction enzyme resulted in a high transformation efficiency, with up to 67% of transformants containing a single copy of transforming DNA. In contrast to protoplast transformation, electroporation of spores in the presence of a restriction enzyme did not improve transformation efficiency or lead to insertion at genomic restriction sites. Southern analysis indicated that for both protoplast transformation with REMI using KpnI or XhoI and for electroporation of spores without addition of restriction enzymes, transforming DNA inserted at different genomic sites in a high proportion of transformants.     

On the method-independent lower bound (MILB) to the variance of attenuation estimation in random reflection

Based on the Cramer-Rao inequality, the MILB is calculated for the problem of estimating the frequency slope of the attenuation coefficient of tissue from random reflections of ultrasonic waves. Under typical signal conditions, this bound for a 1-cm x 1-cm region was found to be about 0.08 db/(MHz-cm), rather close to the clinical requirement of 0.1 dB/(MHz-cm). Comparison to existing methods (including an autoregressive deconvolution method) shows room for further improvement.     

Bilateral polynomial matrix equations in two indeterminates

Two special cases of the bilateral 2-D polynomial matrix equationDU +VN=C whenC=I andC=I with being a -stable 2-D polynomial, which are related respectively to deadbeat and asymptotic control problems of 2-D systems, are first considered. By generalizing the concepts of factor coprimeness, zero coprimeness and zero skew primeness in the 2-D polynomial ring to the ring of causal -stable 2-D rational functions, a constructive solution of these two problems mentioned is proposed. Based on these results, we derive a solvability condition for the bilateral equiation whereC is a general 2-D polynomial matrix. The general solutions are investigated as well.     

Highly reliable a‐IGZO TFTs on a plastic substrate for flexible AMOLED displays

We have successfully reduced threshold voltage shifts of amorphous In–Ga–Zn–O thin‐film transistors (a‐IGZO TFTs) on transparent polyimide films against bias‐temperature stress below 100 mV, which is equivalent to those on glass substrates. This high reliability was achieved by dense IGZO thin films and annealing temperature below 300 °C. We have reduced bulk defects of IGZO thin films and interface defects between gate insulator and IGZO thin film by optimizing deposition conditions of IGZO thin films and annealing conditions. Furthermore, a 3.0‐in. flexible active‐matrix organic light‐emitting diode was demonstrated with the highly reliable a‐IGZO TFT backplane on polyimide film. The polyimide film coating process is compatible with mass‐production lines. We believe that flexible organic light‐emitting diode displays can be mass produced using a‐IGZO TFT backplane on polyimide films.     

Vivid image projection system using e‐Paper active screen

Projected image on a screen is not always vivid enough when it is projected in a bright room. We generally have to choose either vivid image in a dark room or dull image in a bright room. We have suggested a new projection system which can realize high contrast image projection in a bright room. It is consisted of electronic paper screen, projector, and light‐emitting diode room light. The electronic paper (e‐Paper) screen alternates its whole surface white/black with 120 Hz. Room lights are controlled with the same frequency of 120 Hz with the opposite phase as that of the e‐Paper screen. We have confirmed that our new system has achieved higher contrast than the conventional projection system and also enough readability of paper documents in the room simultaneously. We have thus confirmed that our new system can bring an ideal condition, for audience, which can realize a vivid image projection in a bright room.