Assessment of rabbit spleen size using ultrasonography

Assessment of spleen size using the ultrasonography has become a standard practice in human. However, the assessment is not established method in experimental animals. To establish the index to assess the spleen size using ultrasonography, we measured the cross-section image of rabbit spleen during endotoxin shock. The image of the cross-section was appeared as triangle, and the height of the triangular image was defined as the spleen index. This spleen index showed strong correlation with the spleen weight. In conclusion, this method is suitable for observation of changes in rabbit spleen size and may reduce the number of rabbit in the longitudinal studies.     

Transformation of trifoliate orange with rice chitinase gene and the use of the transformed plant as a rootstock

RCC2, cDNA clone encoding rice class-I chitinase, was introduced into trifoliate orange (Poncirus trifoliata Raf.). Chitinase activity of the transformed plants was higher than that of the non-transformed plants. The introduced gene was not detected in the scions grafted onto the transformed plants. The effect of the introduction of foreign gene into rootstocks on the scions was discussed.     

Comparative Evaluation on Nitrogen Saturation of Forest Catchments in Japan and Northeastern United States

To analyze the differences in the status and processes of nitrogen saturation in Japan and northeastern United States, we examined the hydrobiogeochemistry of nitrogen of forested watersheds in these regions. Two distinct differences were found between watersheds in Japan compared with those in US. 1) In Japanese watersheds, marked decreases of NO₃ ^? concentration in surface waters during the summer growing season were not found and NO₃ ^? concentrations sometimes increased especially in the summer at nitrogen saturated sites. This contrast with watersheds in US where decreases in NO₃ ^? concentration during the summer are commonly observed except those watersheds in advanced stages of nitrogen saturation. These differences in NO₃ ^? concentration relationships can be attributed to climatic differences, with Japan having high precipitation and high discharge during the summer, while in many regions of North America lowest discharges are found in the summer. The climatic regime in Japan leads to high rates of mineralization and the rapid transport of NO₃ ^? to streams in summer. 2) Japanese watersheds, even those with high NO₃ ^? concentrations in surface waters, show little evidence of acidification. This is in contrast to sites in US where increased NO₃ ^? concentrations, especially during episodic events, result in surface water acidification.     

Novel preparation method of template DNAs from wine for PCR to differentiate grape (Vitis vinifera L.) cultivar

Template DNAs were extracted from wine and purified for use as samples for PCR to differentiate grape cultivars. It has been pointed out that the authentication of grape material by PCR using wine as a material is very difficult. The problems are (1) decomposition of DNAs during fermentation; (2) contamination of DNAs from microorganisms such as yeast; (3) interference of DNA extraction by polysaccharides and polypeptides in the beverages; and (4) coexistence of PCR inhibitors, such as polyphenols. For this study was developed a novel preparation method of template DNA from wine to differentiate grape cultivars using PCR by (1) lyophilizing and pulverizing the fermented beverage to concentrate the DNAs; (2) decomposition of polysaccharides and proteins so as not to inhibit DNA extraction using heat-resistant amylase and proteinase K without DNA damage by endogenous DNase; and (3) separation of the template DNAs for PCR from PCR inhibitors, such as polyphenols, by purification using 70% EtOH extraction and isopropyl alcohol precipitation. To prevent the amplification of microorganisms' DNAs during PCR, suitable PCR primers closely related to the specific plant DNAs, such as chloroplast DNA and mitochondrial DNA, were selected. The sequences of the amplified DNAs by PCR were ascertained to be the same as those of grape materials.     

Biomass allocation and nitrogen limitation in a Cryptomeria japonica plantation chronosequence

We investigated soil net nitrogen mineralization rate, above- and belowground biomass allocation, and nitrogen use in a Cryptomeria japonica plantation chronosequence. Total biomass accumulation showed an asymptotic accretion pattern, and the peak total biomass accumulation rate occurred approximately 30 years after afforestation. Soil net nitrogen mineralization rate was lowest 30 years after afforestation. Between years 30 and 88, net nitrogen mineralization increased again. These results indicate that an imbalance in soil nitrogen supply and plant nitrogen demand occurred approximately 30 years after afforestation. Furthermore, leaf nitrogen concentration, which was used as an index of plant nitrogen status, was lower in mature forest than in young forest, suggesting that mature stands did not take up nitrogen as successfully. If soil resources such as nitrogen limit plant growth, plants may increase biomass allocation to fine root structure; however, fine root biomass was not higher in 30- and 88-year-old stands than in younger stands, suggesting that changes in biomass allocation may not be effective against nitrogen deficiency in a C. japonica plantation chronosequence.     

Vascular smooth muscle lipofuscinosis occurring predominantly in veins of a cynomolgus monkey (Macaca fascicularis)

A 6-year-old male cynomolgus monkey showed chronic wasting. No gross abnormalities were observed in necropsy except for changes secondary to wasting. Microscopic examination revealed pigment granules deposition in systemic smooth muscles. They were observed as brown or basophilic in hematoxylin and eosin stain, and were positive for periodic acid-Schiff, Schmorl and Ziehl-Neelsen. Ultrastructurally, they consisted of residual bodies surrounded with varying amounts of solitary ribosomes. Thus, these granules were considered as lipofuscin. Unlike brown bowel syndrome in humans, the pigment granules were distributed systemically not only in the digestive tract but also in the blood vessels predominantly in the veins. To our knowledge, this is the first report on vascular smooth muscle lipofuscinosis occurring predominantly in the veins of primates.     

Isolation and tyrosinase inhibitory effects of polyphenols from the leaves of persimmon, Diospyros kaki

The main polyphenols were isolated from the leaves of six selected persimmon cultivars. Seven compounds were obtained by reverse-phase HPLC, and their structures were elucidated by multiple NMR measurements. These compounds are hyperoside, isoquercitrin, trifolin, astragalin, chrysontemin, quercetin-3-O-(2'-O-galloyl-β-D-glucopyranoside) (QOG), and kaempferol-3-O-(2'-O-galloyl-β-D-glucopyranoside) (KOG). Their inhibitory activity was tested against tyrosinase for the oxidation of L-DOPA, and only chrysontemin showed inhibitory activity. To investigate the differences of their inhibitory effects, the tyrosinase inhibitory activities of their aglycons, cyanidin, quercetin, and kaempferol, were also tested. As a result, it was confirmed that the most influential moiety for tyrosinase inhibition was the 3',4'-dihydroxy groups of the catechol moiety. Moreover, the tyrosinase inhibitory activity of chrysontemin, which was identified in persimmon leaves for the first time, is supported by a simulated model of chrysontemin docking into mushroom tyrosinase.     

Methane flux characteristics in forest soils under an East Asian monsoon climate

Methane (CH₄) uptake by soil can possibly be suppressed more in regions with heavy summer precipitation, such as those under the East Asian monsoon climate, as compared to that in regions with a dry summer. In order to determine how precipitation patterns affect seasonal and spatial variations in CH₄ fluxes in temperate forest soils, such fluxes and selected environmental variables were measured on different parts of a hill slope in a cypress forest in central Japan. On the upper and middle parts of the slope, CH₄ uptake was observed throughout the year, and the uptake rates increased slightly with soil temperature and decreased with soil water content. The CH₄ flux predicted using data for the middle and upper parts of the slope ranged from −1.12 to −0.83 kg-CH₄ ha⁻¹ y⁻¹ (i.e. CH₄ uptake by soil) and from −2.30 to −2.04 kg-CH₄ ha⁻¹ y⁻¹, respectively. In contrast, in the relatively wet lower part of the slope near an in-stream wetland, large CH₄ emissions (>2 mg-CH₄ m⁻¹ d⁻¹) were observed during the rainy summer. In this wetter plot, the soil functioned as a net annual CH₄ source in a rainy year. Hence the variation in CH₄ flux with a change in soil water conditions and soil temperature on the lower part of the slope contrasted to that on the upper and middle parts of the slope. The predicted CH₄ flux for this lower plot ranged from −0.45 kg-CH₄ ha⁻¹ y⁻¹ in a dry year to 1.80 kg-CH₄ ha⁻¹ y⁻¹ in a rainy year. Our results suggest that consideration of the soil water conditions across a watershed is important for estimating the CH₄ budgets for entire forest watershed, particularly in regions subject to a wet summer.     

Importance of basic cultivation techniques to increase irrigated rice yields in Tanzania

The productivity of irrigated rice is low in Tanzania. We hypothesized that this is caused by the absence of a packaged application of basic cultivation techniques. A baseline survey of 31 rice irrigation schemes across the country revealed that a large proportion of fields were cultivated without a technical package. Thus, a package was introduced to each of the 31 schemes through a farmer-to-farmer (FTF) extension approach. First, selected key farmers (KFs) were trained with the basic cultivation techniques at agricultural training institutes. Second, the KFs transferred their knowledge to intermediate farmers (IFs) by working together at a demo-field established in each scheme. Third, the KFs and IFs exhibited the rice performance to other farmers (OFs). The paddy yield across the 31 schemes greatly increased from the pre-training level of 2.4 t ha^?1 to 3.6 t ha^?1after the FTF extension. However, the farmer interviews in the four selected schemes suggested that the technical package was not adopted by all farmers owing to the time-consuming nature of the FTF extension. It was inferred from our study that the low productivity of irrigated rice is caused by the absence of basic cultivation techniques in Tanzania. However, the post-training yield remained relatively low compared with high-yields (4.3–8.4 t ha^?1) recorded in cultivar selection trials and high-performing schemes in the county. This “yield gap” could be partly ascribed to the insufficient technical diffusion and the technique-dependent adoption among OFs.