Formation of callose from sucrose in cotton fiber microsomal membranes

Callose is formed from exogenous sucrose by cotton fiber microsomal membranes that contain both sucrose synthase and callose synthase activity. Although the coupled reaction between sucrose and callose synthases occurs predominantly to channel glucose from sucrosederived uridine diphosphate (UDP)-glucose into callose in the membranes, there is no difference in the UDP-glucose-forming/sucrose-forming activity ratios between the soluble and membrane-bound forms of sucrose synthase. The consumption of UDP-glucose from sucrose into callose probably leads to UDP-glucose formation rather than sucrose formation despite the lower affinity of sucrose synthase for sucrose than for UDP-glucose. Callose formation is markedly stimulated by the addition of either recombinant Glu¹¹ (S11E) or in vitro phosphorylated Ser¹¹ mung bean sucrose synthase but not by the wild-type nonphosphorylated Ser¹¹ enzyme. We propose that a negative charge (by phosphorylation or mutagenesis) at Ser¹¹ in sucrose synthase causes the enzyme to promote a coupled callose-forming reaction.Part of this paper was presented at the 8th International Cell Wall Meeting, Norwich, UK, September 1998     

Shear tests of timber joints composed of sugi composite glulam beams using newly developed steel connectors

A research project supported by the Japan Wood Working Machinery Association has been conducted since 1999 to examine the feasibility of sugi (Cryptomeria japonica D. Don) composite glulam beams reinforced with Douglas fir (Pseudotsuga menziesii Franco) lamination. This study, part of the project, was concerned with the strength properties of timber joints composed of composite glulams using newly developed structural steel connectors. Two types of beam were prepared: 10 plies (inner 6 plies sugi, outer 4 plies Douglas fir) and 8 plies (inner 4 plies sugi, outer 4 plies Douglas fir). Two types of structural steel connector, Haratec and Standard, were used for joining the beam with a post or a girder. The relation between load and deformation of the joints was represented as a typical nonlinear curve. Initial stiffness and maximum load of the joint composed of the composite glulam were in the range between those of sugi and Douglas fir. Strength properties of the joints varied with three variables: type of connector, depth of the glulams, and the type of joint. Thus, the allowable loads for the connectors should be determined for each combination of these variables.     

Orally administered conjugated linoleic acid ameliorates allergic dermatitis induced by repeated applications of oxazolone in mice

Conjugated linoleic acid (CLA) is one of the constituents of animal products with possible health benefits such as anti‐carcinogenic and anti‐obesity effects. In this study, we investigated the immunomodulatory effects of CLA using a mouse model of allergic dermatitis. Mice were orally administered either a CLA mixture containing equal amounts of 9c, 11 t‐CLA and 10 t, 12c‐CLA, or high linoleic acid safflower oil, and allergic dermatitis was induced on the ear by repeated topical applications of oxazolone. Oral administration of the CLA mixture but not the high linoleic safflower oil attenuated the symptoms of allergic dermatitis in both ear weights and clinical scores. This effect was associated with decreased levels of ear interleukin‐4 (IL‐4) and plasma immunoglobulin E. The immunomodulatory effects of the CLA isomers were compared by an in vitro cytokine production assay. The results showed that 9c, 11 t‐CLA, the most predominant isomer in animal products, significantly inhibited IL‐4 and interferon‐γ production from mouse splenocytes with similar potency to 10 t, 12c‐CLA. These findings suggest that CLA, a constituent of animal products, has a potentially beneficial effect for amelioration of allergic dermatitis.     

Effect of oral administration of L-pipecolic acid on food intake in chicks and mice

It has been demonstrated that L-pipecolic acid (L-PA), a major metabolic intermediate of L-lysine (L-Lys) in the brain, is involved in the functioning of the γ-aminobutyric acid-ergic system. Previous study has shown that intracerebroventricular injection of L-PA suppressed feeding and induced sleep-like behavior in neonatal chicks. The present study examines whether the action of L-PA was induced by gavage in both chicks and mice. Oral administration of L-PA significantly inhibited food intake at 2 h after treatment in neonatal chicks, although no significant effect of L-Lys was detected. In mice, oral L-PA suppressed food intake compared to the control after 2 h of treatment. It was concluded that L-PA was effective for suppression of food intake after oral administration in both avian and mammalian species.     

Regulation of elicitin-induced ethylene production in suspension-cultured tobacco BY-2 cells

INF1 elicitin, a proteinaceous elicitor produced by Phytophthora infestans, induces a hypersensitive response in tobacco BY-2 cells. In response to elicitin, tobacco cells produce both reactive oxygen species (ROS) and ethylene (ET). To investigate the regulation of elicitin-induced ET production, we pharmacologically analyzed the effects of several chemicals on ET production. Inhibitors of ROS generation or ROS chelators efficiently inhibited ET production, whereas simultaneous treatment of a superoxide anion-generating system with salicylhydroxamic acid recovered ET production. In an in vitro experiment, superoxide anion was necessary and sufficient for conversion of 1-aminocyclopropane-1-carboxylate (ACC) to ET because ET was produced from ACC solely in the presence of the superoxide-generating chemical KO₂. ET production was also inhibited by lipoxygenase (LOX) inhibitors, indicating a possible involvement of LOX-mediated generation of superoxide anion and ET production itself. Furthermore, elicitin-induced ET production was completely inhibited by the protein synthesis inhibitor cycloheximide but recovered after exogenous application of ACC, indicating that de novo protein synthesis is required for ACC accumulation, leading to ET production. We also investigated the effects of several phytohormones on elicitor-induced ET production and discuss their role in the defense response.     

A pea NTPase, PsAPY1, recognizes signal molecules from microorganisms

Apyrases (E.C.3.6.1.5; NTP-NDPases) are distributed in the cytosol, nuclei, cytoskeleton, and on the surface of plant cells. Some may play an important role in signal transduction from exogenous stimuli. We previously found a protein of ca. 55-kDa (CWP-55) in an ATPase-rich fraction from the pea cell wall bound to the elicitor and supprescins (suppressors of defense) from pea pathogen Mycosphaerella pinodes. We cloned the cDNA of CWP-55 that coincided with PsAPY1, one of two NTPase clones in a pea cDNA library. An analysis with a green fluorescent protein fusion protein indicated that PsAPY1 was distributed in the cell wall, nucleus, and cytoplasm. The recombinant PsAPY1 expressed in Escherichia coli had ATP-hydrolyzing activity responsive not only to the elicitor and supprescins from the pea pathogen but also to other elicitors such as a bacterial harpin, a yeast extract, and a synthetic glycopeptide. Biotinylated fungal signal molecules were bound to the recombinant PsAPY1 specifically. Resonant mirror detection confirmed such binding characteristics of PsAPY1. Based on these results, we discuss the role of cell-wall-bound NTPases in recognizing and responding to microorganisms on the cell wall surface.     

Importance of AC-rich Element on Pea Phenylalanine Ammonia-Lyase Gene 1 Promoter for Expression Induced by Nonpathogenic Attack

Regulatory elements in the promoter of phenylalanine ammonia-lyase gene 1 of pea (PSPAL1) in response to nonpathogenic attack were identified by in vivo footprinting analysis. The footprints determined AC-rich sequences, Box-I and Box-II, that were conserved at similar positions in the phenylpropanoid gene promoters from several plants. To reveal the functions of the AC-rich sequence in nonpathogen-responsiveness, we constructed Box-I-deletion PSPAL1 promoter (dB-1) with GUS reporter gene and transformed it into tobacco plant. The dB-1 had reduced basal expression and a complete loss of nonpathogen-responsiveness. These results indicate the essentiality of Box-I for PSPAL1 activation induced by nonpathogenic attack. Received 27 October 1999/ Accepted in revised form 25 November 1999     

Nitro-PAHs and PAHs in Atmospheric Particulate Matters and Sea Sediments in Hiroshima Bay Area, Japan

Atmospheric and sea sediment concentrations were measured for eight nitrated polycyclic aromatic hydrocarbons (nitro-PAHs) and three unsubstituted PAHs in a suburban area and sea sediments in the Hiroshima Bay watershed area, Japan, from July to December, 2006 (atmospheric particulate matter) and in September and November, 2004 (sea sediments). Atmospheric concentration was higher in winter than summer for both nitro-PAHs and PAHs. Concentrations in sea sediments were less than 10%, and pattern was similar to those of atmospheric particles. Several combustion emission sources were also measured, and the 1-NP/Pyr ratio was compared to environmental values. The ratio of atmospheric and sea sediments were significantly lower than diesel particulate matters. Further, the vehicle emission loading and sea sedimentation loading was evaluated in this watershed area, and from the comparison, the existence of other important sources PAHs were suggested.