Oxidative metabolism of tetrachlorocyclohexenes,pentachlorocyclohexenes, and hexachlorocyclohexenes with microsomes from rat liver and house fly abdomen

Microsomal mixed-function oxidase systems from rat liver and house fly abdomen effectively metabolized isomers of 3,4,5,6-tetrachlorocyclohexene, 1,3,4,5,6-pentachlorocyclohexene, and 1,2,3,4,5,6-hexachlorocyclohexene to tetrachlorocyclohexenol isomers, 2,4,5,-trichlorophenol, and 2,3,4,6-tetrachlorophenol, respectively. The $\text{(}\text{346}\text{5}\text{)-isomer}$ of pentachlorocyclohexene gave also an abundant amount of pentachlorocyclohexenol isomers. As the metabolites of ($\text{36}\text{45}$)-, ($\text{35}\text{46}$)-, and $\text{(}\text{34}\text{56}\text{)-hexachlorocyclohexene}$, some compounds such as 1,2,4-trichlorobenzene, 1,2,3,4-tetrachlorobenzene, and pentachlorobenzene were more abundantly formed, respectively, than 2,3,4,6-tetrachlorophenol. These oxidative metabolic reactions were shown to mainly proceed via “ene-like” hydroxylation accompanied by double bond migration. Inhibition by CO, piperonyl butoxide, and SKF 525-A suggested that the “ene-like” hydroxylating enzyme was cytochrome P-450 dependent. The formation of an isomer of pentachlorocyclohexenol from $\text{(}\text{36}\text{45}\text{)-hexachlorocyclohexene}$ was also observed, and this reaction was activated by SKF 525-A.     

Comparative insecticidal activities and metabolic rates of lindane and its hexadeuterio-analog

Hexadeuterio-lindane(γ-BHC-d₆) was several times as toxic as lindane against the mosquito (Culex pipiens pallens), house fly (Musca domestica), German cockroach (Blattella germanica) and American cockroach (Periplaneta americana). The neuroexcitatory activity of these two compounds did not differ. Lindane was considerably synergized by piperonyl butoxide, but lindane-d₆ was not. A large isotope effect was observed in the in vivo breakdown of lindane-d₆. Thus, the intrinsic toxicities of both compounds are equivalent. The difference in insecticidal activity seems to be due to the different rates of detoxifying biodegradation caused by the kinetic deuterium isotope effect.     

Identification of QTLs controlling harvest time and fruit skin color in Japanese pear (Pyrus pyrifolia Nakai)

Using an F₁ population from a cross between Japanese pear (Pyrus pyrifolia Nakai) cultivars ‘Akiakari’ and ‘Taihaku’, we performed quantitative trait locus (QTL) analysis of seven fruit traits (harvest time, fruit skin color, flesh firmness, fruit weight, acid content, total soluble solids content, and preharvest fruit drop). The constructed simple sequence repeat-based genetic linkage map of ‘Akiakari’ consisted of 208 loci and spanned 799 cM; that of ‘Taihaku’ consisted of 275 loci and spanned 1039 cM. Out of significant QTLs, two QTLs for harvest time, one for fruit skin color, and one for flesh firmness were stably detected in two successive years. The QTLs for harvest time were located at the bottom of linkage group (LG) Tai3 (nearest marker: BGA35) and at the top of LG Tai15 (nearest markers: PPACS2 and MEST050), in good accordance with results of genome-wide association study. The PPACS2 gene, a member of the ACC synthase gene family, may control harvest time, preharvest fruit drop, and fruit storage potential. One major QTL associated with fruit skin color was identified at the top of LG 8. QTLs identified in this study would be useful for marker-assisted selection in Japanese pear breeding programs.     

Distribution of new methylene blue injected into the dorsolumbar epidural space in cows

Objective To describe the effect of injection volume and anatomy on the spread of new methylene blue (NMB) injected into the epidural space between the first and second lumbar vertebrae in cows. Study Design Prospective experimental study. Sample Population Thirteen nonpregnant cows. Methods Cows were randomly assigned to two groups. Group 1 received 5 mL and Group 2 received 10 mL of 0.12% NMB in 0.9% saline. The injection was made into the first interlumbar epidural space using a dorsal approach. The extent of cranial and caudal migration of the dye, as manifested by the staining of the epidural fat and dura mater, was measured. Results Mean ± SEM number (range) of stained vertebrae was significantly greater in the 10‐mL group than in the 5‐mL group, 4.4 ± 0.6 (T11 to L5) and 3.0 ± 0.2 (T12 to L3), respectively (p < 0.05). Linear regression analysis showed that the volume was significantly correlated with the number of stained vertebrae (R² = 0.42, p = 0.016). In the dorsal and lateral aspect of the spinal cord, there were two types of distribution of NMB along the surface of the epidural fat: between the periosteum and epidural fat; and between the epidural fat and dura mater. Migration under the spinal cord occurred along the two longitudinal epidural veins. Conclusions and Clinical Relevance The larger the volume of solution injected into the first interlumbar epidural space, the greater the spread. Intrinsic anatomic factors, such as characteristics of epidural fat and veins, influence the epidural spread of injected solution and, consequently, epidural analgesia.     

Development of an SSR marker set for efficient selection for resistance to black spot disease in pear breeding

Black spot disease, which is caused by Alternaria alternata (Fries) Keissler Japanese pear pathotype, is one of the most harmful diseases in Japanese pear cultivation. Because of the potential harm of fungicides to consumers and the environment, resistant cultivars are desired. In this study, to enable efficient marker-assisted selection in pear breeding, we conducted comprehensive inoculation tests and genotyping with 207 pear cultivars. We identified a marker set (Mdo.chr11.27 and Mdo.chr11.34) suitable for selection for black spot resistance. In most susceptible cultivars, Mdo.chr11.27 amplified a 220-bp band and Mdo.chr11.34 amplified a 259-bp band. The genotype of Mdo.chr11.34 corresponds perfectly to the estimated genotype of Japanese pears susceptible to black spot disease. Using linkage analysis, we identified the positions of the gene for susceptibility to black spot disease in Chinese pear. Mdo.chr11.27 and Mdo.chr11.34 were tightly linked to susceptibility in Chinese pear, and the susceptibility gene was mapped at the top of linkage group 11, similar to that in Japanese pear. This marker set and the accumulation of phenotypic data will enable efficient marker-assisted breeding for black spot resistance in pear breeding.     

Direct genotyping of single pollen grains of a self-compatible mutant of Japanese pear (Pyrus pyrifolia) revealed inheritance of a duplicated chromosomal segment containing a second S-haplotype

Radiation mutant 415-1, which is the first known diploid pollen-part self-compatible mutant of pears (Pyrus spp.), has a decreased ability to produce pollen. To determine whether the self-compatibility trait is associated with this defect, we directly analyzed the genotypes of individual pollen grains by using polymerase chain reaction amplification of DNA from single pollen grains. We isolated single pollen grains from 415-1 and succeeded in genotyping the S-RNase gene and three simple sequence repeat (SSR) markers in linkage group 17. Out of 173 individual pollen grains, 28 (16 %) were S-heteroallelic. These pollen grains had two alleles each of the S-RNase gene and of two linked SSR loci, all on a duplicated chromosomal segment, but only one allele of a non-duplicated locus farther away on the same chromosome. The segregation ratio of each marker in the pollen from 415-1 was approximately the same as that observed in outcross progeny. This suggests that the decrease in frequency of pollen with the duplicated S-haplotype occurred during meiosis or pollen formation, but that the probability of fertilization by S-heteroallelic pollen is equal to that of single-allelic pollen. However, the partial sterility in 415-1 can also be attributed to one or more unidentified lethal mutations unlinked to the duplicated segment encompassing the S-haplotype. Single-pollen genotyping can be used in a variety of applications in genetic research because in cases where all pollen genotypes are proportionately represented in the progeny, segregation ratios can be obtained without producing the next generation.     

Reovirus isolation from healthy dogs in Japan

Twenty-two strains of viruses isolated from apparently healthy dogs were identified as reoviruses on the basis of their biological, physico-chemical and morphological properties. The cross hemagglutination-inhibition test revealed that all of them were of type 1, except one which was of type 3. The significance of reovirus isolation from healthy animals is discussed.     

Phylogenetic relationships between the lettuce root rot pathogen Fusarium oxysporum f. sp. lactucae races 1, 2, and 3 based on the sequence of the intergenic spacer region of its ribosomal DNA

The genetic relationship between the vegetative compatibility groups (VCGs) and between physiological races of Fusarium oxysporum f. sp. lactucae (FOL), the causal pathogen of lettuce root rot, was determined by analyzing the intergenic spacer (IGS) region of its ribosomal DNA. A total of 29 isolates containing a type strain were tested: 24 Japanese isolates, 2 Californian isolates, and 3 Italian isolates. Three races (races 1, 2, and 3) were found in Japan, and race 1 was also distributed in California and Italy. Races 1, 2, and 3 each belonged to a distinct VCG: VCG-1, VCG-2, and VCG-3 (VCG-3-1, VCG-3-3), respectively. Phylogenetic (neighbor-joining) analysis of the IGS sequences revealed that races 1, 2, and 3 coincided with three phylogenetic groups (PG): PG-1, PG-2, and PG-3, respectively. These results indicate that the three races are genetically quite different and have a strong correlation with VCGs and phylogenetic groupings. The nucleotide sequence data reported are available in the DDBJ/EMBL/GenBank databases under the accession no. AB195218     

Consecutive changes in serum alkaline phosphatase isoenzyme 3 activities in Holstein heifers during the first 18 months of life

This study investigated consecutive fluctuations in serum activities of bone-specific alkaline phosphatase (ALP) isoenzyme 3 (ALP3) in 11 clinically healthy Holstein heifers during the first 18 months of life. ALP3 activities at the first sampling time point after weaning (3 months) were significantly lower than those at multiple time points during the pre-weaning period. Those activities increased from a minimum at 3 months to a peak at 6 months during the post-weaning period. In the anthropometric data, daily body weight and wither height gains appeared to be below the public data at 4 months and 4–5 months, respectively. The data suggested that serum ALP3 activity can be used to monitor skeletal growth of heifers at weaning.