Restriction endonuclease analysis of canine herpesviruses isolated in Japan

Eighteen canine herpesvirus (CHV) isolates from Japan and two reference strains were compared by restriction endonuclease analysis technique using total DNA extracts from cells infected with the viruses. In order to select the suitable restriction endonucleases for differentiation of CHV isolates, ten enzymes were used and three of them, HindIII, XbaI, and PvuII, were found to be useful for strain differentiation. With these enzymes, CHV isolates from unrelated individuals were readily differentiated from each other. In contrast, all the isolates derived from the same litter were not distinguishable on the basis of restriction cleavage patterns. However, slight mobility shifts were observed among the isolates from the same litter or the same individual. The results showed that this method provides a powerful tool for epidemiological surveys of CHV infection.     

An enzyme-linked immunosorbent assay for detection of antibodies to exogenous avian leukosis virus

A microplate enzyme-linked immunosorbent assay (ELISA) for detecting antibodies to avian leukosis virus (ALV) of subgroups A and B in infected chickens was developed with the use of Rous-associated virus (RAV)-1 (subgroup A) and RAV-2 (subgroup B) antigens purified by sucrose-gradient centrifugation. The antigen was used for ELISA after treatment with Triton X-100. In the ELISA, the subgroup viral antigen reacted strongly with homologous antiserum but also reacted with heterologous antiserum. Tests with serum absorbed with purified homologous and heterologous virus and tests for antigen-blocking by group-specific antibodies to ALV revealed that the reaction was caused mainly by subgroup-specific antibodies. The ELISA was 8 to 32 times more sensitive than the virus-neutralization (VN) test and detected antibodies to ALV earlier than the VN test in chickens infected experimentally with RAV-1 and RAV-2. In field application of the ELISA, 44.2% of 484 chicken sera were positive for RAV-1 and/or RAV-2 antigen, and 80.4% of flocks were positive. These findings indicate that ELISA is superior to the VN test in sensitivity, simplicity, rapidity, and applicability for large-scale field surveys for ALV infection.     

Serotypes and antimicrobial susceptibility of Pseudomonas aeruginosa strains isolated from diseased dogs

Strains of Pseudomonas aeruginosa isolated from diseased dogs in Tokyo area during 1983 through 1986 were serotyped and assayed for antimicrobial susceptibility to obtain an epizootiological aspect of canine P. aeruginosa infection. Major sources of specimens were ear and nasal discharges and urine. The results of O-antigen typing using a monoclonal antibody kit showed that the most predominant serotype was type M. Types G and B were also major serotypes. In a yearly distribution of serotypes, type I was almost limited in 1986, and was isolated mainly from surgical wounds, which showed an episode of nosocomial infection, whereas that of type M or B was dispersed during 4 years from 1983 to 1986. As a result of antimicrobial susceptibility test, most canine strains were considered to be susceptible to 4 drugs, which were commonly used in both human and veterinary clinics, in contrast to human isolates.     

Extractives relating to heartwood color changes in sugi (<Emphasis Type="Italic">Cryptomeria japonica</Emphasis>) by a combination of smoke-heating and UV radiation exposure

 Sugi green logs with red or black heartwood were smoke-heated, and the changes in the color of the heartwood after ultraviolet (UV) (λ = 365 nm) radiation exposure were then observed. After UV radiation exposure, the redness and yellowness increased in both the red and black heartwoods, whereas the brightness decreased. In the black heartwood, the resulting color turned from yellowish white to reddish brown. Reddening in black heartwood after exposure to a combination of smoke heating and UV radiation is thought to be due to a decrease in brightness and an increase in both redness and yellowness. However, the degree of change in heartwood color by UV radiation exposure was not greatly affected by smoke-heating treatments of various lengths. When methanol extracts were fractionated and exposed to UV radiation, the yellowness increased in the n-hexane-soluble portion and the redness increased in the acetone-soluble fractions from the n-hexane-insoluble portion. These results suggest that the n-hexane-soluble fraction contains the substances that allow heartwood color to change to yellow after UV radiation exposure, and the acetone-soluble-fraction from the n-hexane-insoluble portion contains the substances that allow it to change to red. Received: November 14, 2001 / Accepted: June 3, 2002 Acknowledgment This research was supported in part by a Grant-in-Aid for Scientific Research from the Japan Society for the Promotion of Science. This study was presented in part at the 51st Annual Meeting of the Japan Wood Research Society, Tokyo, April 2001 Correspondence to:N. Yoshizawa     

Synthesis of dihydroxyphenacyl glycosides for biological and medicinal study: β-oxoacteoside from Paulownia tomentosa

The protected structure of -oxoacteoside (tomentoside A), 2-oxo-2-(3,4-dihydroxyphenyl)ethyl 3-O-(2,3,4-tri-O-acetyl--l-rhamnopyranosyl)-4-O-caffeoyl--d-glucopyranoside 14 was synthesized in 14% overall yield in 11 steps, starting from d-glucose for biological and medicinal studies of phenylpropanoid glycosides. The first step was the preparation of a 3-O-rhamnopyranosyl disaccharide sugar core 2 from a suitably protected rhamnosyl trichloroacetimidate 10 and glucose derivative (diacetone-d-glucose 1) in 71% yield. To the glucose moiety of this sugar core, several protection/deprotection procedures were performed sequentially to obtain a fully acetylated sugar core 7 with a 4-OH group on the glucose moiety, in 57% yield in five steps. Thereafter, to the 4-OH group of the glucose moiety, selective 4-O-caffeoylation was achieved by proton-transfer esterification with 3,4-di-O-allylcaffeic acid 16 to give the caffeoyl disaccharide 11 in 97% yield. Then, it was converted to trichloroacetimidate 13 for a glycosylation donor in 90% in two steps. Finally, anomeric glycosylation was conducted with 2-oxo-2-(3,4-di-allyloxyphenyl)ethyl alcohol 19 with catalytic amounts of BF₃·Et₂O to give 2-oxo-2-(3,4-di-allyloxyphenyl)ethyl 2,6-di-O-acetyl-3-O-(2,3,4-tri-O-acetyl--l-rhamnopyranosyl)-4-O-(3,4-di-allyloxycaffeoyl)--d-glucopyranoside 14 in 60% yield. Deprotected intermediates of compounds 2, 11, 14, and 19 which were obtained in high yield would be useful for biological and medicinal studies of phenylpropanoid glycosides.Part of this study was presented at the 52nd Annual Meeting of the Japan Wood Research Society, Gifu, April, 2002     

Generation and alteration of norlignans in a transition zone during the drying of a Cryptomeria japonica log

Sugi (Cryptomeria japonica D. Don) produces secondary metabolite norlignans in xylem. Several norlignans are involved in the coloration of heartwood and defense of sapwood against microbial invasion. Their biosynthetic process should be well understood so that their properties can be exploited to improve the quality and utility of C. japonica wood. Unfortunately, information on the norlignan biosynthesis is limited because norlignans are mainly synthesized in a particular season in the transition zone (TZ) along with the heartwood formation, and is difficult to study. Although the generation of two norlignans of C. japonica, agatharesinol and (E)-hinokiresinol, has been reported, systems for producing other norlignans are not yet known. To establish a novel norlignan generating system, we examined the changes occurring in norlignans in a TZ during the process of drying a C. japonica log. On the day of felling, the TZ contained agatharesinol and (E)-hinokiresinol, which increased until they reached a maximum on day 40 after felling. Sequirin-C appeared on day 40 and increased to day 70. The generation of sequirin-C in the TZ can be used to investigate the biosynthetic process of heartwood norlignans. This study describes for the first time the changes that occur in the composition of norlignan during the drying of the TZ.