Synthesis and insecticidal activity of benzoheterocyclic analogues of N'-benzoyl-N-(tert-butyl)benzohydrazide: Part 1. Design of benzoheterocyclic analogues

The N'-benzoyl group of N-tert-butyl-N'-benzoyl-3,5-dimethylbenzohydrazide (1) was converted to a series of benzoheterocyclecarbonyl groups in order to investigate the potential usefulness of superimposing a hydrazine insecticide on 20-hydroxyecdysone. A series of analogues with benzodioxole, benzodioxane, benzodioxapine, indole, benzoxazole, benzoxazine or benzothiazole instead of the phenyl group of (1) were synthesized and tested for their insecticidal activity against the common cutworm (Spodoptera litura F). N-tert-Butyl-N'-(3,5-dimethylbenzoyl)-1,3-benzodioxole-5-carbohydrazide and N-tert-butyl-N'-(3,5-dimethylbenzoyl)-2,3-dihydro-1,4-benzodioxine-6-carbohydrazide showed high insecticidal activities, superior to that of (1) and equal to that of the commercial insecticide tebufenozide (RH-5992).     

Synthesis and insecticidal activity of benzoheterocyclic analogues of N'-benzoyl-N-(tert-butyl)benzohydrazide: Part 2. Introduction of substituents on the benzene rings of the benzoheterocycle moiety

A series of N'-benzoheterocyclecarbonyl-N-tert-butyl-3,5-dimethylbenzohydrazide analogues possessing a variety of substituents on the benzene rings of the benzoheterocyle moieties were synthesized and tested for their insecticidal activity. The introduction of a methyl group at the R1 position of the benzoheterocycle moiety strongly increased the insecticidal activity. Among the analogues synthesized, N'-tert-butyl-N'-(3,5-dimethylbenzoyl)-5-methyl-6-chromanecarbohydrazide showed the highest insecticidal activity (LC50 = 0.89 mg litre(-1)).     

Systemic spindle-cell proliferative disease in broiler chickens

The major organs and tissues of 24 broiler chickens (70 or 71 days old) suspected of spindle-cell proliferative disease (SPD) because of showing the tumorous lesions distributed throughout the body at meat inspection were collected for histopathological and immunohistochemical examination. Macroscopically, liver, spleen and cecal tonsil showed severe enlargement and white nodules or plaques were observed in the liver, spleen, kidney, intestine and bone marrow of the femur. All chickens were diagnosed with SPD based on the histopathological examination. The lesions of SPD were observed in the liver, spleen, kidney, heart, lung, pancreas, proventriculus, gizzard, duodenum, jejunum, ileum, rectum, cecal tonsil, bursa of Fabricius, bone marrow of the femur and skin. Hemangioma was observed in the lung of 1 bird. Eight 1-day-old specific pathogen-free chicks were inoculated intraperitoneally with 0.25 ml of a 20% homogenate of the affected spleens of three naturally occurring cases. One inoculated bird, necropsied at 10 weeks of age, macroscopically had a white nodule in the kidney and histopathologically had spindle-cell proliferative lesions, a pattern similar to that seen in the naturally occurring cases, in the liver, spleen, kidney, heart, lung, pancreas, proventriculus, duodenum, cecal tonsil and bone marrow of the femur, and was diagnosed with SPD. Immunohistochemically, significant positive reactions with a rabbit antiserum against avian leukosis virus antigens were detected in all spindle cells in the proliferative lesions of all examined SPD cases and in tumor cells of the hemangioma of a field case.     

An antagonistic rhizoplane bacterium Pseudomonas sp. strain EC-S101 physiologically stresses a spinach root rot pathogen Aphanomyces cochlioides

We observed that an antagonistic rhizoplane bacterium Pseudomonas sp. strain EC-S101 induces excessive lateral and apical branching in the hyphae of a root rot phytopathogen Aphanomyces cochlioides AC-5 resulting in radial growth inhibition of hyphae in a dual culture assay. Confocal laser scanning microscopic observations using fluorescent stains indicated an increased quantity of nuclei and lipid bodies in the affected hyphae during the early stage (less affected hyphae) at day 3 of interaction. At a more advanced stage (severely affected hyphae) at day 3, nuclei became smaller and round-shaped compared with the oval shape in AC-5 control hyphae. After 7 days, nuclei disintegrated, and the nuclear materials were released into the disorganized cytoplasm. With transmission electron microscopy at 5 days of interaction, we found that the cell walls of AC-5 hyphae were considerably thicker than those of the control. Enlarged vacuoles, lipid bodies sunk into vacuoles, and vacuoles filled with electron-dense material, followed by an invagination of the AC-5 hyphal cell wall, were commonly observed. Nonmembranous electron-transparent inclusion bodies irregular in size were often distributed in the affected hyphae. By integrating our observations, we conclude that antagonistic effects evoked by strain EC-S101 resulted in the death of AC-5 hyphae, which might contribute to the suppression of A. cochlioides AC-5-linked damping-off disease in its host plants. The nucleotide sequence data reported are available in the DDBJ/EMBL/GenBank databases under the accession number AB190286     

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.     

Acute vascular and interstitial rejection following renal allograft transplantation in dogs

Renal allograft transplantation was performed in four beagles. Immunosuppressive treatment using cyclosporine, mizoribin and prednisolone was continued from Day 5 pre- until Day 20 post-transplantation. Between Days 28 and 32 post-transplantation, an abrupt elevation of the serum creatinine values followed by the development of uremia was seen in all recipients. Histopathology of the allografts examined between Days 28 and 37 revealed edema, necrosis, hemorrhage and severe diffuse interstitial cellular infiltration as well as tubulitis. Glomerular changes notably included swelling of the tufts due to hypercellularity, which was consistent with transplant glomerulitis. The intrarenal arteries exhibited fibrinoid necrosis of the walls and intimal or transmural cellular infiltration. These renal lesions were consistent with those of acute vascular and interstitial rejection in humans.     

Microscopic Studies on Attachment and Differentiation of Zoospores of the Phytopathogenic Fungus <Emphasis Type="Italic">Aphanomyces cochlioides</Emphasis>

The mode of aggregation, attachment and differentiation of zoospores of the phytopathogenic fungus Aphanomyces cochlioides when interacting with the host and a host-specific attractant and a G-protein activator, mastoparan, was studied by light and scanning electron microscopy. When a zoospore approached very close to the host root, it seemed to halt, then coiled its anterior flagellum on its body. The halted zoospore appeared to contact the host surface with its posterior flagellum, which gradually drew the encysting zoospore onto the root surface. The spore then docked precisely on the root surface at its ventral face with the help of the posterior flagellum and anchored itself by releasing some adhesive materials. The adherent spore became a spherical after shedding its flagella and rapidly turned into an expanded cyst forming a smooth cyst coat around it, and finally changed into a smaller cystospore covered with a wrinkled surface. In contrast, the mastoparan- or cochliophilin A-stimulated zoospores on artificial membranes aggregated by using their posterior flagella before encystment. These contrasting phenomena suggest that A. cochlioides zoospores may use their posterior flagella for successful docking on the host surface or for aggregation of encysting spores in the absence of the host. Received 30 August 2001/ Accepted in revised form 8 November 2001     

In situ ruminal degradation and in vitro fermentation characteristics,and antioxidative activities of the lotus rhizome

We evaluated the lotus rhizome as a potential ruminant feed by investigating its compositional properties, in situ degradation profile and in vitro fermentation characteristics with ruminal microbes, in comparison with cereal grains (corn, barley and wheat). The antioxidative activities in the lotus rhizome were also estimated. The soluble fraction of dry matter in lotus tuber was >70%, which was higher than those in the grains. The insoluble fraction in lotus tuber was not degraded by ruminal microbes in accord with a first‐order reaction. In an in vitro experiment, lotus tuber showed lower fermentation at 8 hr compared to the grains, but exhibited higher productions of gas and VFA at 48 hr along with a lower lactate and higher pH. The lower value of final lactate production in lotus tuber, indicating the metabolic capacity for lactate utilization retained, suggests a lower risk of ruminal acidosis compared to grains. Lotus rhizome had high antioxidant activities, with the foliar bud showing the strongest ferric reducing antioxidant power, followed in order by the apical bud, node, residual tuber, edible tuber, and nodal root. For ruminants, the lotus rhizome could thus be not only an energy feed but also the source of natural antioxidants.