Histamine downregulates CD14 expression via H2 receptors on human monocytes

Lipopolysaccharide (LPS) binds to LPS-binding protein (LBP) in plasma and is delivered to the cell surface receptor CD14 on human monocyte. LPS is transferred to the transmembrane signaling receptor toll-like receptor (TLR) 4. In the present study, the effect of histamine on the expression of CD14 on human monocytes was investigated. Histamine concentration- and time-dependently decreased the expression of cell surface CD14, whereas histamine did not decrease mRNA for CD14 nor increase soluble CD14 (sCD14). The inhibitory effects of histamine on CD14 expression were antagonized by H2-receptor antagonist, but not by H1 and H3/H4 antagonist. The effects of selective H2-receptor agonists, 4-methylhistamine and dimaprit, on CD14 expression mimicked that of histamine indicating that histamine regulated CD14 expression through the stimulation of H2-receptors. The pretreatment with histamine partially inhibited the LPS-induced TNF-alpha production in human peripheral blood mononuclear cells (PBMC). Such inhibition might be due to the down-regulation of CD14 expression on monocytes by histamine.     

Genistein,a protein tyrosine kinase inhibitor,suppresses the fusogenicity of Moloney murine leukemia virus envelope protein in XC cells

Summary. XC cells are highly susceptible to syncytium formation by infection of ecotropic murine leukemia viruses (MLVs) and by expression of their envelope protein (Env). By this property, XC cells are widely used to determine titers of ecotropic MLVs. Number of plaques resulted from the syncytium formation in XC cells by ecotropic MLV infection is corresponding to number of the viral particles. XC cells had been established from a v-src-induced rat tumor. It has been reported that transformed cells are more sensitive to Mo-MLV-induced syncytium formation than non-transformed cells. To assess whether the transformation by v-src oncogene in XC cells is involved in the high sensitivity to ecotropic MLV-induced syncytium formation, XC cells were treated with genistein, a protein tyrosine kinase inhibitor. Genistein suppressed the syncytium formation between XC cells and ecotropic Env-expressing 293T cells. This result indicates that protein tyrosine kinase activity is associated with the high sensitivity of XC cells to ecotropic Env-induced syncytium formation.Received January 8, 2003; accepted May 26, 2003     

Rapid identification of metallothionein isoforms in liver cytosol fraction by capillary zone electrophoresis using EDTA

Identification of metallothionein (MT) isoforms on capillary zone electrophoresis (CZE) analysis was studied using a linear polyacrylamide-coated capillary at pH 7.4 and EDTA. The CZE system was able to separate standard (purified and commercially available) MT specimens into their isoforms within 10 min. The peaks of MT-1 and MT-2 isoforms disappeared on addition of EDTA to the specimen, and the disappearance was shown to be time-dependent and dose-dependent, although the reason why the peaks decreased is still unclear. A heat-treated cytosol fraction prepared from Zn-injected mouse liver showed many major and minor peaks on CZE analysis. Two major peaks were identified to be MT-1 and MT-2, respectively, by co-injection with the purified MT isoforms. When EDTA was added to the cytosol fraction, the two major peaks, MT-1 and MT-2, and three other minor peaks disappeared time-dependently. Therefore, each MT isoform in the cytosol fraction can be identified by the addition of EDTA, also the peaks are identified by the corresponding migration times of purified MTs. Unknown substances like MT sub-isoforms may also be detected, although this question warrants clarification. From these results, it was concluded that the addition of EDTA is useful for identification of MT isoforms in cytosol fractions on CZE analysis.     

Comparison of high-energy photon and electron dosimetry for various dosimetry protocols

The American Association of Physicists in Medicine Task Group 51 (TG-51) and the International Atomic Energy Agency (IAEA) published a new high-energy photon and electron dosimetry protocol, in 1999 and 2000, respectively. These protocols are based on the use of an ion chamber having an absorbed-dose to water calibration factor with a 60Co beam. These are different from the predecessors, the TG-21 and IAEA TRS-277 protocols, which require a 60Co exposure or air-kerma calibration factor. The purpose of this work is to present the dose comparison between various dosimetry protocols and the AAPM TG-51 protocol for clinical reference dosimetry of high-energy photon and electron beams. The absorbed-dose to water calculated according to the Japanese Association of Radiological Physics (JARP), International Atomic Energy Agency Technical Report Series No. 277 (IAEA TRS-277) and No. 398 (IAEA TRS-398) protocols is compared to that calculated using the TG-51 protocol. For various Farmer-type chambers in photon beams, TG-51 is found to predict 0.6-2.1% higher dose than JARP. Similarly, TG-51 is found to be higher by 0.7-1.7% than TRS-277. For electron beams TG-51 is higher than JARP by 1.5-3.8% and TRS-277 by 0.2-1.9%. The reasons for these differences are presented in terms of the cavity-gas calibration factor, Ngas, and a dose conversion factor, Fw, which converts the absorbed-dose to air in the chamber to the absorbed-dose to water. The ratio of cavity-gas calibration factors based on absorbed-dose to water calibration factors, N60Co(D,w), in TG-51 and cavity-gas calibration factors which are equivalent to absorbed-dose to air chamber factors, N(D,air), based on the IAEA TRS-381 protocol is 1.008 on average. However, the estimated uncertainty of the ratio between the two cavity-gas calibration factors is 0.9% (1 s.d.) and consequently, the observed difference of 0.8% is not significant. The absorbed-dose to water and exposure or air-kerma calibration factors are based on standards traceable to the National Institute of Standards and Technology (NIST). In contrast, the absorbed-dose to water determined with TRS-398 is in good agreement with TG-51 within about 0.5% for photon and electron beams.     

Spontaneous and osmium tetroxide-induced mutagenesis in an Escherichia coli strain deficient in both endonuclease III and endonuclease VIII

Thymine glycol, uracil glycol, 5-hydroxycytosine and 5-hydroxyuracil are common base lesions produced by cellular metabolism as well as ionizing radiation and environmental carcinogens. Escherichia coli DNA glycosylase, endonuclease III and endonuclease VIII recognize and remove these lesions from DNA. In this study, we assessed the mutagenic potential of these lesions in the supF gene as a forward mutation target in double-stranded plasmid DNA using an E.coli strain deficient in both endonuclease III and endonuclease VIII. These lesions were introduced into pTN89 DNA by the chemical oxidant osmium tetroxide. Spontaneous supF mutations occurred at a frequency of 3.03x10(-7) and osmium tetroxide-induced at a frequency of 8. 25x10(-7). Sequence analysis of supF mutants revealed that mutations occurred at cytosine sites rather than thymine sites, suggesting that thymine glycol is not the principal premutagenic lesion. In contrast, G:C-->A:T transitions were dominantly detected in the spontaneous and osmium tetroxide-induced mutations in the endonuclease III and endonuclease VIII double defective host. In this case, products of cytosine oxidation such as 5-hydroxycytosine, which are the substrate for endonuclease III and endonuclease VIII, were the principal mutagenic lesions.     

The inhibitory effect of oxatomide, an anti-allergic agent, on eosinophil-mediated and eosinophilic cell line-mediated natural cytotoxicity against bronchial epithelial cells

Recently, eosinophils have been implicated as inflammatory effector cells in allergic and inflammatory reactions such as bronchial asthma. In this study eosinophil-mediated and eosinophilic cell line-mediated natural cytotoxicity against bronchial epithelial cells and the effects of oxatomide, an anti-allergic agent, on their cytotoxicity were investigated. Treatment with oxatomide diminished both eosinophil-mediated and eosinophilic cell line-mediated natural cytotoxicity in vitro. We concluded from these results that oxatomide not only has anti-allergic activity but also anti-inflammatory properties for eosinophils. In addition this method for isolating eosinophils seems to well serve the purpose of evaluating eosinophil function as in this investigation, as we have reported previously.