Development of a simple IgE‐independent anaphylactic model using buckwheat antigen and B‐type CpG oligodeoxynucleotide from Streptococcus thermophilus

We developed a severe anaphylactic model in mice using buckwheat antigen and B‐type CpG‐oligodeoxynucleotides (CpG‐ODNs) from Streptococcus thermophilus genome. In typical systemic anaphylaxis models, animals are challenged with large quantity of antigens via an intravenous (i.v.) route. Here, we showed a simple anaphylactic shock after challenge via intraperitoneal (i.p.) route. The i.p. method is simpler than i.v. administration and has a lower risk for failure. To generate this anaphylactic model, 5‐week‐old female BALB/c mice were first i.p. sensitized with buckwheat antigen mixed with B‐type CpG‐ODN. After 2 weeks, mice were challenged with antigen to induce anaphylactic shock, which was evaluated by scoring the severity symptoms and measuring serum levels of various proteins and splenic cell producing cytokines. Immunoglobulin (Ig)G_2a production and interferon‐γ positive cells were markedly increased in mice immunized with antigen mixed with B‐type CpG‐ODN, whereas serum IgE levels were decreased by B‐type CpG‐ODN. We also examined the effects of various ODNs (A, B and C‐type CpG‐ODNs) and antigens (buckwheat, α‐casein, β‐lactoglobulin and ovalbumin) on anaphylactic severity, and found that the combination of buckwheat and B‐type CpG‐ODN induced the most intense anaphylactic shock. This model is expected to contribute to the study of the prevention of anaphylactic shock.     

Imaging diagnosis--polioencephalomalacia in a calf.

A 2-month-old mix-breed calf developed acute blindness and ataxia. Serum thiamine concentration was deficient. In antemortem magnetic resonance imaging there were laminar T2-hyperintense regions extending along the cerebral cortex that primarily affected the gray matter. The lesions were relatively symmetric between the left and right hemispheres but no abnormalities were present at the frontal lobes. At necropsy, laminar autofluorescence of the cerebral cortex was observed under ultraviolet exposure at 365 nm, consistent with a diagnosis of polioencephalomalacia. Polioencephalomalacia in the bovine species is compared with that in other species, namely humans, dogs, and cats.     

Gene expression profiling in embryonic chicken ovary during asymmetric development

The reproductive system in female birds arises as bilateral asymmetrical anlagen, excluding the birds of prey. Earlier, histological and messenger RNA (mRNA) expression profile studies of several genes related to gonadal sex differentiation in chicken embryos tried to elucidate the query of this asymmetry in a scattered manner. To understand the matter precisely, we have focused on mRNA expression of a cohort of genes (FSHR, CYP19A1, caspase 3, caspase 8) in second half of the embryonic days (E10–E18). The established role of leptin in development of the embryo and its expression in the embryonic ovary also drove us to check leptin receptor (LEPR) expression in the ovary. Increased expression of FSHR and CYP19A1 in the left ovary compared with that in the right ovary was identified (P < 0.05), promoting preferential left ovarian development and functionality. Significant high expression (P < 0.05) of the apoptotic genes in the right ovary were also involved here. Leptin probably has no direct influence on ovarian asymmetry as no significant variation in gonadal mRNA expression of LEPR was observed within the same experimental days. We propose that asymmetric expression of this cohort of genes (FSHR, CYP19A1, caspase 3, caspase 8) leads to the development of dimorphic gonads during embryogenesis.     

Pool‐based genome‐wide association study identified novel candidate regions on BTA9 and 14 for oleic acid percentage in Japanese Black cattle

Fatty acid composition is an important indicator of beef quality. The objective of this study was to search the potential candidate region for fatty acid composition. We performed pool‐based genome‐wide association studies (GWAS) for oleic acid percentage (C18:1) in a Japanese Black cattle population from the Hyogo prefecture. GWAS analysis revealed two novel candidate regions on BTA9 and BTA14. The most significant single nucleotide polymorphisms (SNPs) in each region were genotyped in a population (n = 899) to verify their effect on C18:1. Statistical analysis revealed that both SNPs were significantly associated with C18:1 (p = .0080 and .0003), validating the quantitative trait loci (QTLs) detected in GWAS. We subsequently selected VNN1 and LYPLA1 genes as candidate genes from each region on BTA9 and BTA14, respectively. We sequenced full‐length coding sequence (CDS) of these genes in eight individuals and identified a nonsynonymous SNP T66M on VNN1 gene as a putative candidate polymorphism. The polymorphism was also significantly associated with C18:1, but the p value (p = .0162) was higher than the most significant SNP on BTA9, suggesting that it would not be responsible for the QTL. Although further investigation will be needed to determine the responsible gene and polymorphism, our findings would contribute to development of selective markers for fatty acid composition in the Japanese Black cattle of Hyogo.     

Immune synergistic oligodeoxynucleotide from Lactobacillus rhamnosus GG enhances the immune response upon co‐stimulation by bacterial and fungal cell wall components

Bacterial genomic DNA has recently been shown to elicit a highly evolved immune defense. This response can be selectively triggered for a wide range of therapeutic applications, including use as a vaccine adjuvant to immunotherapies for allergy, cancer, and infectious diseases. Previously, we identified a low‐concentration immune synergistic oligodeoxynucleotide (iSN‐ODN, named iSN34) from Lactobacillus rhamnosus GG that has immunosynergistic activity upon costimulation of target cells with ligands of Toll‐like receptor 9 (TLR9). Here, we extend that observation by demonstrating the synergistic induction (in mouse splenocytes) of IL‐6 by the combination of iSN34 with cell wall components of bacteria and fungi. We observed that splenocytes pretreated with iSN34 and then costimulated with agonists for TLR1/2 (Pam₃CSK₄), TLR4 (lipopolysaccharide), or TLR2/6 (Zymosan) exhibited enhanced accumulation of IL‐6. These results suggested that the combination of iSN34 with TLR1/2, TLR4, or TLR2/6 agonists may permit the induction of a potent immune response.     

Transfer of the Rsv3 locus from ‘Harosoy’ for resistance to soybean mosaic virus strains C and D in Japan

Resistance to soybean mosaic virus (SMV) is imperative for soybean (Glycine max (L.) Merr.) production in the Tohoku region. Molecular markers for SMV resistance were previously reported for U.S. SMV strains, but they cannot be applied because of the differences in strain classification between Japan and the U.S. A U.S. variety ‘Harosoy’ has been used mainly as a donor of resistance to SMV strains C and D in a Japanese breeding program, resulting in resistant varieties such as ‘Fukuibuki.’ Because ‘Harosoy’ harbors the Rsv3 gene conferring resistance to the virulent SMV strain groups, G5 through G7, it appears that the Rsv3 gene confers resistance to strains C and D. In this study, we introduced resistance to the two strains from ‘Fukuibuki’ into a leading variety ‘Ohsuzu’ by recurrent backcrossing with marker-assisted selection. All lines selected with markers near Rsv3 showed resistance to the strains, suggesting that the Rsv3 locus is responsible for the resistance. Three years of trials showed that one of the breeding lines, ‘Tohoku 169,’ was equivalent to ‘Ohsuzu’ with respect to agricultural characteristics such as seed size, maturity date, and seed yield, except for the SMV resistance.     

Predicting chronosequential changes in carbon stocks of pachymorph bamboo communities in slash-and-burn agricultural fallow,northern Lao People’s Democratic Republic

In northern Lao People’s Democratic Republic, rising human population has drastically reduced the fallow period of slash-and-burn agriculture which has led to a considerable decrease in the carbon stock in these communities. We estimated chronosequential changes in the communities' carbon stocks, and established the relationship between the fallow period and fallow-period-average carbon stocks in three carbon pools of bamboo-dominated communities in hilly areas of the Luang Prabang Province, northern Lao People’s Democratic Republic. Based on measurements by destructive sampling, we devised a model and root-to-shoot ratios for estimating bamboo biomass. In six secondary plant communities established after slash-and-burn cropping, we estimated community biomass using the above model and others, and measured deadwood and litter stocks. The communities’ biomass and deadwood significantly increased with time after the last cropping and the former reached about 100 Mg ha⁻¹ after 15 years, whereas litter stocks did not show significant trends over time. Extending the fallow period from 2 to 5 years would increase fallow-period-average carbon stock from 14.2 to 25.1 Mg C ha⁻¹. The overstory height was significantly correlated with biomass, deadwood, and litter carbon stocks of these communities. Based on our findings, changes in a community’s carbon stocks can be estimated using the changes in overstory height, which should be taken into account in future studies to reduce uncertainty in estimating carbon stocks in tropical ecosystems.     

Biomass of a man-made forest of timber tree species in the humid tropics of West Java, Indonesia

Biomass of a mature man-made forest in West Java, Indonesia, was estimated to evaluate the carbon sequestration potential of plantation forest in the humid tropics. Twenty plots, each 0.25 ha in area and containing one to six planted species over 40 years of age and with closed canopies, were selected. Trunk dry mass was estimated from trunk diameter, tree height, and bulk density. Maximum trunk diameter (122 cm) was observed in a 46-year-old Khaya grandifoliola C. DC. tree, and the tallest tree (51 m) was a 46-year-old Shorea selanica (DC.) Blume. The largest trunk biomass (911 Mg ha⁻¹) was achieved in the plot composed of two Khaya spp. Among the plots composed of indigeneous Dipterocarpaceae species, the largest trunk biomass was 635 Mg ha⁻¹. These trunk biomasses were larger than those reported from primary rainforests in Southeast Asia (e.g., 403 Mg ha⁻¹ in East Kalimantan, 522 and 368 Mg ha⁻¹ in Peninsular Malaysia). The large biomass in this forest suggests that, given favorable conditions, man-made forests can accumulate the quantities of atmospheric carbon that were lost by the logging of primary forests in the humid tropics.     

Individual variation in low boiling point monoterpenes emitted from hinoki (Chamaecyparis obtusa) needles

Variations in the compositions of low-boilingpoint (LBP) monoterpenes in needle samples of 50 hinoki (Chamaecyparis obtusa) trees were investigated using the headspace technique. Considerable compositional variations were revealed, especially in sabinene composition. The sabinene composition varied from 4.9% to 78.0% of the total LBP monoterpenes.-Pinene, myrcene, and limonene also showed considerable variations (9.0%–32.7%, 5.5%–22.6%, 3.6%–29.0% respectively). Analysis of the monoterpene composition allowed definition of four chemotypes based on the contingency table test. No correlation was observed between tree size and LBP monoterpene composition, indicating that the compositional variation in LBP monoterpene exists genetically in this population of hinoki.Part of this paper was presented at the 11th annual meeting of the Chugoku Shikoku branch of the Japan Wood Research Society, Matsue, September 1999