Purification, bacteriolytic activity, and specificity of beta-lytic protease from Lysobacter sp. IB-9374

Lysobacter sp. IB-9374, which was isolated from soil as a high lysyl endopeptidase-producing strain (Chohnanet al., FEMS Microbiol. Lett., 213, 13-20, 2002), was found to produce a beta-lytic protease capable of lysing gram-positive bacteria such as Staphylococcus aureus, Microccocuseus, and Bacillus subtilis. The Lysobacter strain secreted the beta-lytic protease into the culture medium at a 2.4-fold higher level than Achromobacter lyticus. The enzyme was highly purified through a series of six steps with a high yield. The enzyme was strongly inhibited by tetraethylene-pentamine and 1,10-phenanthroline. The purified enzyme lysed more efficiently almost all the gram-positive bacteria tested than lysozyme, lysostaphin, and mutanolysin. The enzyme was very similar to Achromobacter beta-lytic protease containing one zinc atom in terms of amino acid composition and N-terminal sequence. The nucleotide sequence revealed that the mature enzyme was composed of 179 amino acid residues with additional 198 amino acids at the amino-terminal end of the enzyme. The deduced amino acid sequence of the mature enzyme coincided with that of the Achromobacter enzyme, although the prepro-region showed a 41% sequence identity with the counterpart. These results indicate that Lysobacter sp. is a useful strain for an efficient large-scale preparation of beta-lytic protease capable of lysing bacteria.     

Apoptosis Signaling Is Altered in CD4(+)CD25(+)FoxP3(+) T Regulatory Lymphocytes in Pre-Eclampsia

The aim of our study was to estimate the surface expressions of CD95 (APO-1/Fas) antigen and the intracellular expressions of anti-apoptotic protein Bcl-2 and pro-apoptotic protein Bax in CD4(+)CD25(+)FoxP3(+) T regulatory lymphocytes (Tregs) as well as the percentage of CD8(+)CD28(+) T cytotoxic cells in peripheral blood of patients with pre-eclampsia in comparison with healthy pregnant women in the third trimester of physiological pregnancy. Twenty-four women with pre-eclampsia and 20 normal third trimester pregnant women were included in the study. The lymphocytes were isolated from peripheral blood samples and labeled with monoclonal antibodies. The expressions of surface antigens and intracellular proteins were estimated using flow cytometry. The population of CD4(+)CD25(+)FoxP3(+) Treg cells was significantly lower in peripheral blood of patients with pre-eclampsia when compared to normal third trimester pregnant women. The percentages of CD4(+)CD25(+)FoxP3(+) Treg cells that express Bcl-2 protein were significantly lower in peripheral blood of patients with pre-eclampsia when compared to healthy pregnant women, whereas the percentages of CD4(+)CD25(+)FoxP3(+) Treg cells with the expressions of Bax protein did not differ in both groups. Moreover, the mean fluorescence intensity (MFI) of Bcl-2 protein in CD4(+)CD25(+)FoxP3(+) Treg cells was significantly lower and MFI of Bax protein significantly higher in pre-eclampsia when compared to the control group. The percentage of CD8(+)CD28(+) T cells did not differ in both studied groups but MFI of CD28 antigen on T CD8(+) cells was significantly higher in pre-eclampsia when compared to the control group. The obtained results suggest that the deficit of CD4(+)CD25(+)FoxP3(+) Treg lymphocytes which is observed in pre-eclampsia may be associated with altered apoptosis signaling in Tregs.     

A multiplexer with coupled quarter wave distributed lines in highdielectric ceramics

A multiplexers available several purposes such as for mobile communications, CATV systems and multimedia systems in future, are propped and trially made by authors. This paper present the results the design concept and experimental results. The design principle was based on the theoretical concept proposed by us     

Studies on catalysis by molten metals. 14. Catalytic cracking of asphalts at low temperature

The catalytic effects of liquid metals for low-temperature (336 °C) asphalt-cracking have been examined using a semi-batch reactor. All the liquid metals examined (Bi, Cd, Ga, In, Pb and Sn) effectively catalysed the reduction of molecular weight with minimal gasification (<1 wt%). Gasification had a positive correlation with the reduction in molecular weight in the processed asphalts. The catalytic activities of liquid metals tor these two reactions were proportional to a parameter which represents the interaction between the atoms of the catalyst metal and the radicals formed in the course of the reaction. Mass flow among four fractions (saturates, aromatics, resins and asphaltenes) had two independent paths: conversion of resins to saturates and of aromatics to asphaltenes. The former path dominated when catalytic dehydrogenation activity was relatively low, whereas the latter dominated when the catalytic activity was high.     

Absorption and Fluorescence Spectra of Nitrobenzanthrones

To identify nitrobenzanthrones (NBAs) such as 3-nitro-7 H -benz[ de ]anthracene-7-one (3-NBA) on the basis of their electronic spectral data, we have synthesized 1-, 2-, 3-, 9-, and 10-NBAs and measured their absorption and emission spectra. The first strong absorption band of the NBAs appeared in the region 350-440 nm; the band of 10-NBA was red-shifted by about 20 nm. The molar absorptivities of 3- and 9-NBA were about two times as large as those of the others. The fluorescence spectra of the NBAs varied more largely with the position of the nitro group compared to the absorption spectra. The quantum yields of fluorescence were very small, ranging from ～10 m 2 for 2- and 10-NBA to ～10 m 4 for 1- and 3-NBA. 3-NBA exhibited a characteristic spectrum with two broad bands at 450 and 530 nm. On excitation causing an n ~ * transition in the NBAs, their fluorescence intensities increased by a factor of 10. Phosphorescence was observed at 77 K for all compounds though that of 1-NBA was very weak. These results give the promise of characterizing individual isomers of NBAs on the basis of their spectroscopic data.     

Ultra Low-Dose Radiation: Stress Responses and Impacts Using Rice as a Grass Model

We report molecular changes in leaves of rice plants (Oryza sativa L. - reference crop plant and grass model) exposed to ultra low-dose ionizing radiation, first using contaminated soil from the exclusion zone around Chernobyl reactor site. Results revealed induction of stress-related marker genes (Northern blot) and secondary metabolites (LC-MS/MS) in irradiated leaf segments over appropriate control. Second, employing the same in vitro model system, we replicated results of the first experiment using in-house fabricated sources of ultra low-dose gamma (γ) rays and selected marker genes by RT-PCR. Results suggest the usefulness of the rice model in studying ultra low-dose radiation response/s.     

Brain histamine mediates the bombesin-induced central activation of sympatho-adrenomedullary outflow

Intracerebroventricular (i.c.v.) administration of bombesin (0.3 nmol) increased plasma levels of both adrenaline and noradrenaline in urethane anesthetized rats. These bombesin-induced increases were inhibited by i.c.v. pretreatment with pyrilamine, an H1-receptor antagonist. Ranitidine, an H2-receptor antagonist also inhibited the increase of adrenaline, however, its effective dose was much larger than that of pyrilamine. Furthermore, the bombesin-induced increase of noradrenaline was not effectively inhibited by ranitidine. In the next series, turnover of histamine was assessed by measuring accumulation of tele-methylhistamine (t-MH), a major metabolite of brain histamine. I.c.v. administration of bombesin (0.3-3 nmol) increased turnover of hypothalamic histamine, while its intravenous administration was without effect. The present results suggest that the bombesin-induced central activation of sympatho-adrenomedullary outflow is probably, at least in part, mediated through brain histaminergic neurons.     

A new method of controlling cavity-induced pressure oscillations using sub-cavity

A new passive control technique of cavity-induced pressure oscillations has been investigated numerically for a supersonic two-dimensional flow over open rectangular cavities at Mac h number 1.83 at the cavity entrance. A subcavity on the front wall of the cavity covered by a flat plate was evaluated for the effectiveness of controlling cavity-induced acoustic oscillations. The results showed that sub-cavity is very effective in reducing cavity-induced pressure oscillations. The results also showed that the resultant amount of attenuation of cavity-induced pressure oscillations was dependent on the length and thickness of the flat plate, and also on the depth of the sub-cavity used as an oscillation suppressor.