In vitro hepatic biotransformation of aldrin and dieldrin in food-producing animals.

The hepatic biotransformation of aldrin (AD) and dieldrin (DD) was studied in liver post-mitochondrial supernatants (S-9s) from laying hens, female cattle and swine. S-9s were incubated with 0.03 nmol of AD or DD for 1 h. After 1 h, AD in the samples was almost epoxidated to DD. This formation was found with all the animal S-9s, and the highest rates occurred in pig S-9 (P < 0.01), followed by cow and hen S-9s. No reduction of DD was found with any of the S-9s.     

Measurement of sea bottom surface backscattering strength by quantitative echo sounder

ABSTRACT:   Measurement theory and method of the bottom surface backscattering strength (SS) using a quantitative echo sounder (QES) are discussed and applied in the ocean near Java, Indonesia. The frequencies of the QES were 38, 70, and 120 kHz. The measurements of bottom echoes and sampling of bottom material by a dredge were done simultaneously. Bottom characterization was based on analysis of particle size distribution for bottom samples taken during the survey. The SS value increases with the increase of the mean diameter of the bottom particles. The SS decreased with increasing frequency. The effectiveness of QES for measurement of the SS along with observation of the depth topography were demonstrated.     

S-allele genotypes of apple pollenizers,cultivars and lineages including those resistant to scab

Summary

We examined S-allele genotypes of ten apple cultivars and species to determine their possible usefulness as pollenizers for all apple cultivars. ‘Dolgo’ did not contain any known S-RNases encoded at the S-locus, suggesting its possible usefulness as a pollenizer for almost all apple cultivars. We also identified and confirmed the S-allele genotypes of 18 apple cultivars by polymerase chain reaction (PCR)-digestion analysis. The S-genotype of ‘Kiou’ (S₁S₇), ‘Korei’ (S₃S₂₈), ‘Korin’ (S₁S₉), ‘Kotoku’ (S₁S₂₈), ‘Kyokkou (S₇S₂₅), ‘Lobo’ (S₁S₇), ’Mahe 7’ (S₂S₇), ‘Mellow’ (S₂S₃), ‘Takahara’ (S₃S₉) and ‘Warabi’ (S₉S₂₈) were confirmed by pollination results. These cultivars seemed not to have originated from the expected seed or pollen parents or, in the case of ‘Lobo’, might have been mislabelled. Finally, we identified the S-allele genotypes of ‘Prima’ (S₂S₁₀), ‘Querina’ (S₃S₉) and ‘Yoko’ × ‘Prima’ (S₃S₁₀), which are resistant to scab.     

Elimination half-lives of sulphadimethoxine and its N4-acetyl metabolite in tissues of laying hens

The depletion rates of sulphadimethoxine (SDM) and its metabolite N4-acetylsulphadimethoxine (N4-AcSDM) were estimated in blood and various tissues of laying hens. The tissue contents (ppm) of SDM and N4-AcSDM after the withdrawal of SDM, which was fed to hens at 400 ppm diet for 5 successive days, were determined by HPLC. The elimination half-life (t1/2) of N4-AcSDM in the liver, ovary and muscle was estimated to be 4.3 h with a 95% confidence interval from 3.6 to 5.3 h. No significant difference between t1/2 of N4-AcSDM in the tissues and that of SDM (4.4 h) in the blood, kidney, muscle, ovary and adipose tissue was observed. On the other hand, the t1/2 of N4-AcSDM in the kidney (8.1 h) was significantly longer than that in the above 3 tissues.     

Transferring and distributing profiles of p,p'-(DDT) in egg-forming tissues and eggs of laying hens following a single oral administration

Laying hens were administered orally with a single dose of p,p'-(DDT) (1 mg/kg bodyweight). The concentrations (microg/g) of DDT or its metabolites, p,p'-(DDE) and p,p'-(DDD), in the main tissues involved in egg formation (blood, liver, ovary, and oviducts) and egg yolk, collected 1 day after DDT dosing, were determined by normal-phase high-performance liquid chromatography. The limits of detection were 0.04 microg/g for DDT, 0.07 microg/g for DDE and 0.06 microg/g for DDD. In extractable fats from the above tissues and egg yolk, DDT and DDE were transferred/distributed throughout the tissues and egg yolk. DDD was detected only in the liver. The findings indicate that DDT is metabolized instantaneously to DDE/ DDD in the hen's body and they are transferred rapidly into the egg-forming tissues and egg yolk. Among the four tissues and yolk fats examined, the DDT levels were high in the ovary, oviduct and egg yolk; the DDE levels were high in the liver, ovary and oviduct and lowest in the yolk (P < 0.01).     

Genomic organization of the chicken T-cell receptor beta chain D-J-C region

We examined overlapping genomic clones containing the chicken T cell receptor (TCR) Dbeta-Jbeta-Cbeta complex, which contains a single diversity segment, four joining segments and four exons that encode the constant region. This sequence comprised 18.3 kb. All four Jbeta sequences possessed typical recombination signal sequences (RSS) with intervening 12-bp spacers at their 5'-ends and splice sites at their 3'-ends. No Jbeta-pseudogenes were identified. TGTG sequences in the RSS heptamer sequences were well conserved, as is the case in mammals. A chicken repeat 1-like sequence was found in the intron region between Jbeta-1336 and Cbeta, and several small repeat sequences were identified in intron regions throughout this cloned genome. As germline sequences revealed complete Jbeta sequences, the CDR3 (complementarity-determining region) sequences of TCRbeta from non-immunized splenocytes were analyzed. Non-coding (N) and palindromic (P) nucleotides were frequently observed at the Dbeta-Jbeta recombination sites. There were differences in length of deletion at the 5'-end of each Jbeta. Deletion of the 5'-end of Jbeta-1280 was particularly short when compared with that of Jbeta-1336, but there were no changes in the length of the CDR3 using any of the four Jbeta sequences.     

Construction of chicken-mouse chimeric antibody and immunogenicity in mice

Chicken monoclonal antibodies are potentially useful for diagnostic research and have clinical applications, as chicken show higher potential for antibody production with mammalian-conserved biological molecules. However, the applications of chicken antibodies are limited because of their immunogenicity in mammals. To overcome this problem, we have constructed a chicken-mouse chimeric antibody containing the chicken variable region and the mouse constant region. This chimeric antibody retained similar binding affinities as the parental chicken antibody. The chimeric antibody was also producible as an ascitic antibody in BALB/c mice. Furthermore, when the chimeric antibody was administered to mice, it did not provoke the mouse anti-chicken antibody response. These results indicate that the chimeric antibody is suitable for application to preclinical mouse studies.     

In vitro system for immunoglobulin class switching using BHK cells transfected with murine recombinant CD40 ligand.

To develop an in vitro system for mouse immunoglobulin (Ig) class switching, the expression vector of murine CD40 ligand (CD40L) which is expressed on T cells was transfected to BHK cells. By using culture plates coated with the BHK cells expressing the recombinant CD40L, Ig class switching of splenic B cells was examined. The CD40L mRNA was cloned from splenic T cells of BALB/mice activated with anti-CD3 antibody in vitro. As the No.593 base in the open reading frame sequence of the CD40L from BALB/c spleen differed from T to G, when compared with the known sequence from C57BL/6, one of the BALB/c-derived clones was reconstructed to the known CD40L by site-directed mutagenesis. Splenic B cells from BALB/c were induced secretion of Ig isotypes, IgM, IgG1 and IgE when cultured on two types of BHK cells, the transfected BHK cells with a CD40L clone from BALB/c and the transfected BHK cells with the reconstructed CD40L clone, in the presence of IL-4. However, when splenic B cells from C57BL/6 were cultured on the same systems, the B cells produced Ig isotypes, IgM, IgG1, IgG2a, IgG2b, IgG3 and IgE. In the similar experiments using the transfected BHK cells with a empty vector and the normal BHK cells, none of B cells produced any Ig isotypes other than IgM. These results indicate that Ig class switching of murine B cells can be induced by using these two types of CD40L-expressing BHK cells in vitro.