Role of adherent spleen cells in the induction of cytotoxic activity by Toxoplasma lysate antigen.

In order to identify mechanisms responsible for the anti-tumor effects of Toxoplasma lysate antigen (TLA), we used an in vitro 51Cr release assay to study the functional properties of plastic-adherent cells during induction of splenic cytotoxic activity by TLA. Cytotoxic activity of non-adherent cells was measured in all experiments after a 6 days incubation. Induction of cytotoxic non-adherent cells by TLA required the presence of plastic-adherent spleen cells. In contrast, rhIL-2 alone was able to induce transformation of cytotoxic non-adherent cells from non-adherent spleen cells. Contact between adherent and non-adherent spleen cells was necessary for successful induction of cytotoxic non-adherent cells by TLA. Treatment of spleen cells with anti-macrophage serum prevented induction of cytotoxic activity by TLA. Biologically active IL-2 was not detected in culture supernatants of spleen cells exposed to TLA. These findings suggest that contact between TLA-sensitized non-adherent cells and macrophages is necessary for induction of cytotoxic cells in the presence of TLA. This contact, however, is not necessary for generation of IL-2-induced killer cells.     

Identification of the feline CD63 homologue using retrovirus-mediated expression cloning

Recently, we combined a retrovirus-mediated expression cloning with a simple screening method using non-adherent cells and panning [Anal. Biochem. 315 (2003) 138]. In this study, we applied this method to identify the antigen recognized by an uncharacterized monoclonal antibody raised against a feline cell line, and identified it as the feline homologue of CD63. This simple method is useful for characterizing unknown antibodies that recognize cell surface molecules. Furthermore, the monoclonal antibody identified as an anti-feline CD63 antibody will be useful for studying feline molecular function(s).     

The unstable fate of seedlings of the small-seeded pioneer tree species, Betula maximowicziana

The dynamics of the seedling population of the pioneer tree, Betula maximowicziana, were studied for 6 years under various site conditions, with special reference to mortality and mortality factors, in order to better understand their seedling bank formation process. We found that current year seedlings suffered high annual mortality of more than 80%, but that mortality fell with age. Canopy covers and cohorts maintained strong influences on seedling mortality during the first 3 years after emergence. The open site with disturbed soil surface was best suited for 3-year seedling survival. Since the seedlings of B. maximowicziana emerged regardless of canopy covers, high seedling mortality was thought to be the main factor that interrupted their regeneration under a canopy. The occurrence of various hazards together caused high seedling mortality. The major mortality factors, regardless of site conditions, were drought, cold hazard and debilitation, though occurrence of the three factors varied, dependent upon the climate of that particular year. Annual mortality and the occurrence of each mortality factor at a fine scale of 0.2 m² quadrate each showed certain spatio-temporal variations within the same site condition. The extremely high mortality of B. maximowicziana seedlings should be attributed to their inherently small, initial size. The poor root system of the small seedlings may be responsible for their low resistance to various hazards, such as drought and frost heaving.     

Changes in above- and belowground biomass in early successional tropical secondary forests after shifting cultivation in Sarawak,Malaysia

Uncertainties in the rate of biomass variation with forest ageing in tropical secondary forests, particularly in belowground components, limit the accuracy of carbon pool estimates in tropical regions. We monitored changes in above- and belowground biomass, leaf area index (LAI), and biomass allocation to the leaf component to determine the variation in carbon accumulation rate with forest age after shifting cultivation in Sarawak, Malaysia. Nine plots in a 4-year-old forest and fourteen plots in a 10-year-old forest were monitored for 5 and 7 years, respectively. Forest and plant part biomass were calculated from an allometric equation obtained from the same forest stands. Both above- and belowground biomass increased rapidly during the initial decade after abandonment. In contrast, a much slower rate of biomass accumulation was observed after the initial decade. LAI also increased by approximately double from the 4-year-old to 10-year-old forest, and then gently increased to the 17-year-old forest. We also found that allocation variation in leaf biomass and nitrogen was closely related to the rate of biomass accumulation as a forest aged. During the first decade after abandonment, a high biomass and nitrogen allocation to the leaf component may have allowed for a high rate of biomass accumulation. However, reduction in those allocations to leaf component after the initial decade may have helped to suppress the biomass accumulation rate in older secondary forests. Roots accounted for 14.0–16.1% of total biomass in the 4–17-year-old abandoned secondary forests. We also verified the model predicted values for belowground biomass by Cairns et al. (1997) and Mokany et al. (2006), although both models overestimated the values throughout our data sets by 45–50% in the 10-year-old forest. The low root:shoot ratio in the secondary forests may have caused this overestimation. Therefore, our results suggest that we should modify the models to estimate belowground biomass considering the low root:shoot ratio in tropical secondary forests.     

A surge-like increase in luteinizing hormone preceding musth in a captive bull African elephant (Loxodonta africana)

This study was conducted to determine the correlation between reproductive hormones and musth in a male African elephant. Changes in circulating luteinizing hormone (LH), follicle stimulating hormone (FSH), testosterone and immunoreactive (ir-) inhibin and the degree of musth were evaluated for 4 years. LH increased 4 weeks before musth began. The highest concentrations of testosterone and ir-inhibin were observed from April to October. There were positive correlations among testosterone, ir-inhibin and musth behavior. These findings suggested that the surge-like LH in the pre-musth period might stimulate secretion of testosterone and ir-inhibin and thus initiate the musth behavior. This study also suggested that the high LH level before musth might be a useful biomarker for the beginning of the musth season.     

Development of genotyping method for functionally relevant variants of cytochromes P450 in cynomolgus macaques

In cynomolgus macaques (Macaca fascicularis), widely used in drug metabolism studies, CYP2C9, CYP2C76, CYP2D6, CYP3A4, and CYP3A5, important drug‐metabolizing enzymes, are abundantly expressed in liver and metabolize cytochrome P450 substrates. CYP2C9 (c.334A>C), CYP2C76 (c.449TG>A), CYP2D6 (c.891A>G), CYP3A4 (IVS3 + 1G>del), and CYP3A5 (c.625A>T) substantially influence metabolic activity of enzymes, and thus are important variants in drug metabolism studies. In this study, a real‐time PCR method was developed for genotyping these variants. The validity of the methods was verified by genotyping two wild type, two heterozygous, and two homozygous DNAs and was used to genotype 41 cynomolgus macaques (from Cambodia, Indonesia, the Philippines, or Vietnam) for the five variants, along with another important variant CYP2C19 (c.308C>T). The CYP2C9 and CYP2C19 variants were found only in Cambodian and Vietnamese animals, while the CYP2C76 and CYP2D6 variants were found only in Indonesian and Philippine animals. The CYP3A4 and CYP3A5 variants were not found in any of the animals analyzed. Mauritian animals, genotyped using next‐generation sequencing data for comparison, possessed the CYP2C19 and CYP2D6 variants, but not the other variants. These results indicated differences in prevalence of these important variants among animal groups. Therefore, the genotyping tool developed is useful for drug metabolism studies using cynomolgus macaques.     

Effects of Shifting Cultivation on Soil Ecosystems in Sarawak, Malaysia. III. Results of Burning Practice and Changes in Soil Organic Matter at Niah and Bakam Experimental Sites

The effects of burning on the levels of soil organic matter, soil nitrogen, and soil microbial biomass were studied by carrying out experimental shifting cultivation at two sites, Niah and Bakam in Sarawak, Malaysia. Vegetation biomass was burned in plots (10 × 10 m²) at the rates of 0 (control), 100, 200, and 300 Mg ha⁻¹ at the Niah site and 0, 20, and 100 Mg ha⁻¹ at the Bakam site. At the Niah site, the levels of total C and N of the soils did not change throughout the experiment in spite of enhanced soil respiration until 2 months after burning. Although burning induced an increase in the amount of NH₄-N of the soils, the readily available pool of N (the sum of the NH₄-N, NO₃-N, microbial biomass N, and extractable organic N pools) in the burned plots was depleted appreciably at the end of rice cultivation. The effects of burning on these properties tended to be substantial with increasing amounts of the vegetation biomass burned. On the other hand, the levels of total C and N and the readily available N pool at the Bakam site were low before burning compared with those at the Niah site, and the burning treatments did not affect them appreciably. While the rice yield at the Niah site reached the average value obtained in traditional shifting cultivation in Sarawak, that at the Bakam site was much lower. It was suggested that the flush of NH₄-N induced by burning was one of the major factors for rice growth.