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.     

Decreased expression of matrix metalloproteinases and tissue inhibitors of metalloproteinase in the kidneys of hereditary nephrotic (ICGN) mice

Matrix metalloporoteinases (MMPs), which are dominantly regulated by tissue inhibitors of metalloproteinase (TIMPs), play important roles in extracellular matrix (ECM) degradation and are involved in the progression of kidney diseases. In glomeruli and tubulointerstitum of hereditary nephrotic (ICR-derived glomerulonephritis: ICGN) mouse kidneys, hyper-accumulation of ECM components occurred, and MMP activity decreased. In the present study, because lower levels of MMP activity may contribute to the progression of renal fibrosis in ICGN mice, Western blotting analysis and immunohistochemical staining for MMPs and TIMPs were performed to verify the expression levels of these proteins. Levels of MMP-2, MMP-9, MT1-MMP, TIMP-1 and TIMP-2 in the kidneys were decreased in ICGN mice in comparison with normal ICR mice. These results indicate that small amounts and low levels of activity of MMPs cause the progression of renal fibrosis in ICGN mice.     

Localization of proliferative and apoptotic cells in the kidneys of ICR-derived glomerulonephritis (ICGN) mice

The ICR-derived glomerulonephritis (ICGN) mouse is a novel inbred mouse strain with a hereditary nephrotic syndrome, considered to be a good model of human idiopathic nephrotic syndrome and develops proteinuria, hypoproteinemia and anemia. In the present study, we compared the cell kinetics in the kidneys of ICGN mice with age-matched ICR mice as normal controls. The proliferating cells were visualized by 5-bromo-2'-deoxyuridine labeling, and apoptotic cells were determined by terminal deoxynucleotidyl transferase-mediated biotinylated deoxyuridine triphosphate nick end-labeling. Many proliferating epithelial cells of renal tubules, glomerular mesangial cells and tublointerstitial fibroblast-like cells were observed in the kidneys of ICGN mice, but no proliferating cells were seen in the kidneys of ICR mice. Apoptotic cells had round nuclei, and were observed only in the tubulointerstitium in the kidneys of ICGN mice but not in that of controls. The proliferation of renal tubular epithelial cells may represent a compensatory response, and that of mesangial and fibroblast-like cells may play a pathogenic role in nephrotic syndrome. Apoptosis in tubulointerstitial cells with round nuclei may have been erythropoietin-producing cells, and probably caused anemia.     

Separation of deterrents to ingestive behavior of cattle from cattle feces

Feeding-deterrent chemicals were extracted from cattle feces and then separated with three chromatographic methods. Behavioral two-choice test bioassays with cattle were used to examine the deterrent properties of the fractions. Cattle feces were extracted with diethyl ether, and the extracts were separated into neutral, acidic, and basic fractions. Of the three fractions, only the neutral fraction was a deterrent. Separation of the ether-soluble neutral chemicals was conducted with an open column of silica gel using four carrier solutions consisting of pentane and ether. Fraction B (eluted with the carrier solution; pentane:ether = 90:10) was the most effective deterrent among the four fractions. This fraction was divided into 10 fractions by liquid chromatography. Fractions 6, 7, and 8 seemed to deter cattle from feeding. The combined Fractions 6, 7, and 8 were separated into 15 fractions with HPLC. Deterrent activities were detected in Fractions 2, 3, 9, 10, 11, 12, 13, and 14, suggesting that deterrents were separated into two groups using HPLC. These results suggested that several specific chemicals in feces are involved in inhibiting cattle from ingesting grass near cattle feces.     

Hepatic lesions caused by migrating larvae of Ascaris suum in chickens

Group A consisted of chickens infected with a single dose of Ascaris suum and group B of chickens infected with two successive doses. At days 1, 3, 7, 14 and 21 after the first or second infection dose, six chickens from each group were sacrificed. In both groups, larvae were recovered from the livers on days 1, 3, and 7 and lungs on days 3 and 7. No larvae were detected in chickens on day 14. Clear white lesions were noticed only on the livers from chickens of group B at day 7 but had disappeared at day 14. A comparison with group B showed mild histological changes that developed relative to the livers from group A.     

Abnormalities of extracellular matrices and transforming growth factor beta1 localization in the kidney of the hereditary nephrotic mice (ICGN strain).

ICR-derived strain with glomerulonephritis (ICGN) is a strain of mice with hereditary nephrotic syndrome with an unidentified cause. Based on histopathological and biochemical data, ICGN mice are considered to be a good experimental model for human idiopathic nephrotic syndrome. In the present study, we histochemically investigated the changes in localization of extracellular matrix (ECM) components and transforming growth factor beta1 (TGF-beta1). Strong immunohistochemical staining of basal membrane ECM components (collagen IV and laminin) and interstitial ECM components (type III collagen and fibronectin) were demonstrated in glomeruli and tubulointerstitum of ICGN mice as compared with those of sex and age-matched ICR mice, used as normal healthy controls. Marked type I collagen and tenascin deposition, which were not detected in the glomeruli of ICR mice, were seen in the glomeruli of ICGN mice. A remarkable increase in active-TGF-beta1 was also detected only in glomeruli of ICGN mice, but not in those of ICR mice. Furthermore, strikingly increased alpha-smooth muscle actin, a marker of activated glomerular mesangial cells, was demonstrated in the glomeruli, mainly in the mesangial cells, of ICGN mice. These findings indicated that ECM components are increased in the glomerulus and tubulointerstitum of ICGN mice, and that active-TGF-beta1 induces such increases in ECM components. The present findings may contribute to elucidation of the pathogenic mechanisms of hereditary nephrotic syndrome in ICGN mice and, in future, human idiopathic nephrotic syndrome.     

Pharmacological effects of Ganoderma lucidum collected from ume (Japanese apricot) trees

We performed functional evaluation of the fruiting bodies of Ganoderma lucidum growing on ume trees (Japanese apricot, Prunus mume), and determined the suitability of pruned ume branches as a basic component of culture medium for this mushroom. We observed that all tested functional activities of the fruiting bodies of G. lucidum collected from ume trees were higher than those collected from other broadleaf trees or cultured artificially; the functional tests were angiotensin I-converting enzyme inhibitory activity, a platelet aggregation inhibition test, and an interleukin-8 (IL-8) gene expression inhibition test. When extracts from fruiting bodies of G. lucidum were orally administered to spontaneous hypertensive rats, hypotensive effects were found. Freeze drying was the most suitable procedure for preservation of the extracts, and the activities of 30% ethanol extracts and 30% methanol extracts were higher than those of hot-water extracts. The highest functional activities for extracts from G. lucidum mycelia cultured on sawdust media were for sawdusts based on ume wood. Part of this study was presented at the 56th Annual Meeting of the Japan Wood Research Society, Akita, Japan, August 2006     

城市地下水硝酸盐污染及其成因分析

用N同位素分析方法并结合调查区域土地利用类型的分析 ,对杭州市城区 2 1口水井取样分析以确定杭州城市地下水的水质结果显示 :本地区地下水硝酸盐污染严重 ,杭州城市地下水水质属于Ⅲ类水标准 ,不宜饮用。有 4 0 5 %样品的NO3 N含量超过了世界卫生组织的标准 (N10mgL-1)。我们发现不同的土地利用区有不同的NO3 N水平 (N 0 0 4～ 34 4 1mgL-1)。同时我们引进N同位素方法以辨明NO3 N污染源 :居住区地下水δ15NNO3值为 10 4‰～ 2 2 0‰ ,农业区δ15NNO3值的为 17 5‰～ 19 5‰。生活污水是城市浅层地下水的主要NO3 N污染源。在居住区还存在点源污染 ,如化粪池 ;种植蔬菜施用的有机肥则是农业区的NO3 N污染源。