Characterization of a neutralizing monoclonal antibody against botulinum ADP-ribosyltransferase,C3 exoenzyme

A monoclonal antibody, named C302, was prepared and characterized against botulinum ADP-ribosyltransferase C3 exoenzyme that inactivates RhoA GTP-binding protein, resulting in the neurite outgrowth of human neuroblastoma GOTO cells. C302 bound not to the smaller fragments derived from the protease-treated C3 exoenzyme but to the intact C3 exoenzyme. It seems that the C302 epitope may depend on the three-dimensional structure of C3 exoenzyme molecule. C302 depressed the enzymatic and biological actions of C3 exoenzyme. The dose-dependent depression pattern of C302 on the enzyme activity was similar to that to the biological one. C302 turned the neurite-bearing shape of the C3 exoenzyme-treated GOTO cells into the intact shape. By using of C302 mAb and C3 exoenzyme, the research concerning GTP-binding proteins would be improved.     

Nucleotide sequence of the coat protein gene of Mirafiori lettuce virus

 The coat protein (CP) gene of Mirafiori lettuce virus (MiLV), a tentative member of the genus Ophiovirus was isolated and sequenced. The established sequence consists of 1514 nucleotides including one open reading frame (ORF) with 1311 nucleotides that encodes 437 amino acids with a relative molecular mass 48 543. When the ORF was expressed in Escherichia coli, the obtained protein was confirmed as CP by Western blotting using an antiserum against MiLV. Database searches showed that the CP gene of MiLV has a sequence similar to that of Citrus psorosis virus (CPsV), the type species of the genus Ophiovirus. The comparison between MiLV and CPsV CP genes revealed that the identities of the nucleotide and amino acid sequences were 46.5% and 30.9%, respectively. Received: July 29, 2002 / Accepted: October 2, 2002     

Effects of high ambient temperature and restricted feed intake on urinary and plasma 3-methylhistidine in lactating Holstein cows

Three experiments were performed to examine the effects of high ambient temperature and feed restrictions on urinary 3‐methylhistidine (3MH) excretion and plasma 3MH concentrations as a marker of myofibrillar proteolysis in lactating Holstein cows. In a crossover design, four cows were maintained during two 14‐day treatment periods with ad libitum feed intake under constant moderate (18°C) or high (28°C) ambient temperature (Experiment 1), with ad libitum or 70% ad libitum feed intake under constant moderate temperature (Experiment 2), and with 70% ad libitum feed intake under constant moderate or high ambient temperature (Experiment 3). The total digestible nutrients intake was decreased (P < 0.01) under high ambient temperature (Experiment 1) or feed restriction (Experiment 2). Across experiments, urinary 3MH excretion during days 10–14 of each treatment period was not different among treatments. However, the plasma 3MH concentrations at day 14 of treatment were increased significantly under high ambient temperature with subsequent reduced feeding (P < 0.01) or moderate temperature with feed restriction (P < 0.05), and were increased slightly (P = 0.11) under high ambient temperature alone. These results show that the plasma marker of myofibrillar proteolysis in lactating dairy cows was increased at day 14 of treatments of high ambient temperature with subsequent reduced feeding, moderate temperature with feed restriction, or high ambient temperature alone.     

A Cucumber mosaic virus isolated from Momordica charantia L.

Momordica charantia L. plants systemically infected with Cucumber mosaic virus (CMV) were found in Oita Prefecture. The virus isolated from the host plant was characterized by biological, serological, and molecular biological methods. The purified virus was used to mechanically inoculate the host and produced green mottle, green mosaic, and/or chlorotic spots in the noninoculated upper leaves of the host. The virus was identified as an isolate of CMV containing genomic RNA3 derived from subgroup IA by several lines of evidence based on electron microscopy, serological detection, host range, symptoms, and the entire nucleotide sequence of RNA3.     

Evaluation of beetle capture in traps as compared with manual capture on flowers in a long-term investigation in a beech forest

Beetle samples captured by traps were compared with those captured manually on flowers in a beech forest in Central Japan. Plastic traps (Sankei Chemical Co., Ltd.) equipped with benzyl acetate were set up from the 2nd of June to the 8th of September 1994, and all beetles captured were collected every week. Beetles were also captured on wild flowers blooming along the forest road. The traps captured 21,650 individuals of 169 species in 37 families. On the other hand, 971 individuals of 77 species in 19 families were captured on the flowers. All of the families and 57 (74%) species of the beetles captured on the flowers were also captured by the traps; all of the species with more than five individuals captured on the flowers were also trapped, except for one species. The present trap system is considered to be useful for faunal monitoring of flower-visiting Coleoptera, because the beetle fauna found on wild flowers was virtually covered by the trap capture. A part of this paper was presented at the 106th Annual Meeting of the Japanese Forestry Society (1995).     

Effect of microfibril angle on the longitudinal tensile creep behavior of wood

In this report, we undertook studies of the viscoelastic properties of wood from the viewpoint of the fine structure and properties of the constituent materials in the wood cell wall. To measure the mechanical properties of the wood as the behavior of the cell wall, it is required to perform the longitudinal tensile test using a homogeneous specimen. In this study, microtomed specimens of sugi (Cryptomeria japonica D.Don) earlywood were used for the creep test, which were conducted at the fiber saturation point. The substantial creep compliance of the cell wall was simulated using a simplified viscoelastic model consisting of a Voigt element and an independent spring in series. Based on the experimental results, the values of the parameters were optimized. The results were as follows: (1) the longitudinal tensile creep deformation tends to increase with the elapsed time, similar to the bending creep behavior; (2) the magnitude of the longitudinal creep function increases with MFA; and (3) each parameter in the simplified viscoelastic model is markedly affected by the MFA. Based on these results, the mechanism of the longitudinal tensile creep deformation of wood is discussed.     

Boring activity on coniferous tree branches byXylotrechus villioni (Villard) larvae (Coleoptera: Cerambycidae)

In anAbies homolepis Sieb. et Zucc. forest at Matsumoto, Nagano Pref., cumulative damage byXylotrechus villioni (Villard) (Coleoptera: Cerambycidae) was observed on tree branches. Of the 14 major branches in one sample tree, six had cumulative damage by boring larvae, of which 3 had been bored by multiple larvae. No adult emergence holes were found on the whole tree, suggesting very low survivorship from boring larva to emerging adult. Galleries made by single larvae were often observed to be terminated by a depression apparently carved by a picid woodpecker, suggesting that they are important predators of the cerambycid. The manner of cicatrix formation with phloem tissue over the larval gallery in anAbies veitchii Lindl. branch at Mt. Fuji, Yamanashi Pref. is presented. Studies onXylotrechus villioni (Villard) (Coleoptera: Cerambycidae), a primary borer of coniferous trees in Japan (III). This work was presented at the 104th Annual Meeting of Japanese Forestry Society at Morioka (April, 1993) and 20th International Congress of Entomology at Firenze (August, 1996). This work is supported by a Grant-in-Aid for Scientific Research (No.02660163) from the Ministry of Education, Science, Sports, and Culture of Japan.     

Isolation and sequence analysis of the promoter and an allelic sequence of the iron-sulfur protein subunit gene from the white-rot fungusPleurotus ostreatus

We have isolated a structural gene ofsdil, which encodes the iron-sulfur protein (Ip) subunit of succinate dehydrogenase (EC 1.3.99.1), from a white-rot basidiomycete,Pleurotus ostreatus. Here we report isolation of the promoter region ofsdil and an allelic sequence encoding the second-type cDNA fragment isolated in the former experiments. The nucleotide sequence analysis of the promoter region revealed the existence of putative CAAT and TATA boxes, which permits us to develop an expression system in this species. The Southern blot analysis and the restriction fragment length polymorphism assay using monokaryotic strains demonstrated that no family genes tosdil exist in the haploid genome ofP. ostreatus. Moreover, a genetic analysis to detect a linkage between thesdil genotypes and flutolanil resistance in the mutantP. ostreatus strains was also developed.     

Performances of four chemicals with floral scents as attractants for longicorn beetles (Coleoptera: Cerambycidae) in a broadleaved forest

White plastic insect traps (manufactured by Sankei Chemical Co., Ltd.) equipped with four chemicals with floral scents, namely, benzyl acetate, methyl benzoate, methyl phenylacetate and linalool, were set from June 28 to August 9, 1996. The traps were placed in a Japanese beech (Fagus creata) forest, Minakami, Gunma Prefecture, Central Japan, and trapped longicorn beetles were collected weekly. The greatest number of species and individuals belonged to the subfamily Lepturinae dominated by the genusPidonia. No significant differences were observed in the performances of the above four chemicals in either the total number of species or of individuals. However, more Lamiinae species were captured in the traps with methyl phenylacetate, than in those with methyl benzoate. A part of this paper was presented at the 48th Meeting of the Kanto Branch of the Jpn. For. Soc. (1996).     

Estimating larval supply of Ezo abalone <Emphasis Type="Italic">Haliotis discus hannai</Emphasis> in a small bay using a coupled particle-tracking and hydrodynamic model: insights into the establishment of harvest refugia

Most juveniles of Haliotis discus hannai have been found to be descendants of wild individuals, although most adults were artificially produced (released) individuals as a result of restocking inside the refugium located near the head of Oshoro Bay, Hokkaido, Japan. To estimate the larval supply from released and wild individuals into the refugium and to compare the suitability of locations as larval sources, we simulated larval dispersal using a coupled hydrodynamic and particle-tracking model. The simulation results indicated that more larvae may be supplied from the wild adults inside the bay to the refugium than from the released adults. These results are consistent with the observed high abundance of wild juveniles in the refugium. Most larvae from the refugium were predicted to disperse out of the bay. We found that larval retention in the bay was at least one order of magnitude higher than that in the refugium. Thus, it may be more effective in terms of self-replenishment and reproduction if the refugium were to be expanded to the bay scale. There were only minor differences among the compared sites at the head of the bay in terms of their suitability as larval sources. Consequently, the establishment of new refugia in this area could be expected to provide an effectiveness equal to that of the current refugium.