Genetic characterization of the endangered Kiso horse using 31 microsatellite DNAs

In order to contribute to conservation of the endangered Kiso horse, we clarified their genetic information using 31 microsatellite DNAs, and genotyped 125 horses, 83% of the existing breed. First, we clarified the current status of the horses. The horses were confirmed to have experienced rapid loss of population causing a bottleneck, and their effective population size was much smaller than their census size. Moreover, the number of alleles (6.3), observed heterozygosity (0.674), and expected heterozygosity (0.662) were in the same range as other endangered horses all over the world. Therefore, although their inbreeding level was not so severe (F(is): -0.017), the Kiso horse is surely one of the endangered. Second, we obtained genetic information of individuals. This information allowed us to understand the genetic distance of individuals, and might help in development of a reproductive strategy concerning the genetic distance between the mating pairs. Moreover, there appeared to be 4 subpopulations of Kiso horse, and this result was in good agreement with their historical background. Third, we confirmed that the parentage test for identification using the 31 microsatellite DNAs was highly reliable (probability of exclusion: 0.999999993). This identification increases the reliability of stud certification, and is also helpful for effective management. Understanding the genetic diversity within the population and the relationships among individuals is important to ensuring effective management for maintenance of genetic variation, and this study may help in conservation of the endangered Kiso horse.     

Anti-babesial activity of a potent peptide fragment derived from longicin of Haemaphysalis longicornis

Babesiosis is one of the most important tick-borne diseases affecting livestock that can cause major economic losses worldwide particularly in the tropics. Control relies on controlling both the protozoan parasite and the tick vector. Antiprotozoal drugs are most commonly used for treatment, but problems on emergence of resistant strains and food residues are encountered. Longicin, a defensin-like peptide identified from the hard tick, Haemapysalis longicornis, as well as one of its synthetic partial analogs (P4), were previously reported to exert antimicrobial, fungicidal, and parasiticidal activity. Both longicin and P4 showed babesiacidal activity, in vitro and in vivo. Here, peptide fragments of P4 were studied for in vitro activity against bovine Babesia parasites. One of the peptide fragments, antimicrobial peptide 1 (AMP1), reduced the parasitemia of Babesia bigemina. No peptide had significant effect on Babesia bovis. The sequence of AMP1 corresponded to the longicin sequence which is associated with antiparasitic activity. Although AMP1 caused reduction in parasitemia of B. bigemina, the difference in morphology of the parasite compared with the control group was not statistically significant. However, the percentage occurrence of piroplasms decreased, whereas the abnormal pycnotic form increased. The results demonstrated that this shorter peptide retained the anti-babesial activity of the parent peptide, exerting an antiparasitic effect against a bovine Babesia species. Therefore, this short peptide can be considered for chemical synthesis as an alternative therapeutic agent for babesiosis.     

Production of GM-CSF Mediated by Cysteine Protease of Der f in Canine Keratinocytes

House dust mite (HDM) allergens are the most common allergens for induction of IgE-mediated hypersensitivity. Recently, epicutaneous sensitization with HDM allergens has been emphasized in the development of atopic dermatitis (AD) by producing various soluble factors in keratinocytes. Among the soluble factors, GM-CSF is a key molecule that activates Langerhans cells, antigen-presenting cells in the epidermis. In the present study, we investigated the effects of Dermatophagoides farinae (Der f) on GM-CSF production in a canine keratinocyte cell line, CPEK. CPEKs were found to produce GM-CSF upon stimulation by Der f. The GM-CSF production was suppressed by addition of a cysteine protease inhibitor. The present results suggest that cysteine protease-derived Der f may be an initiator of allergic inflammation by inducing the production of GM-CSF in keratinocytes.     

A novel blast resistance locus in a rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.) cultivar,Chumroo, of Bhutan

The rice cultivar ‘Chumroo’ is commonly cultivated in the mid- and high-altitude areas of Bhutan. This cultivar has shown durable blast resistance in that area, without evidence of breakdown, for over 20 years. Chumroo was inoculated with 22 blast isolates selected from the race differential standard set of Japan. The cultivar showed resistance to all the isolates. To identify the resistance gene(s), Chumroo was crossed with a susceptible rice cultivar, Koshihikari. The F₁ plants of the cross showed resistance. Segregation analyses of 300 F₃ family lines fitted the segregation ratio of 1:2:1 and indicated that a single dominant gene controls the resistance to a blast isolate Ao 92-06-2 (race 337.1). The Chumroo resistance locus (termed Pi46(t)) was mapped between two SSR markers, RM6748 and RM5473, on the terminal region of the long arm of chromosome 4, using linkage analysis with SSR markers. The nearest marker, RM5473, was linked to the putative resistance locus at a map distance of 3.2 cM. At the chromosomal region, no true resistance genes were identified, whereas two field resistance genes were present. Therefore, we designated Pi46(t) as a novel blast resistance locus.     

Dataset of CarboEastAsia and uncertainties in the CO2 budget evaluation caused by different data processing

The datasets of net ecosystem CO₂ exchange (NEE) were acquired from 21 forests, 3 grasslands, and 3 croplands in the eastern part of Asia based on the eddy covariance measurements of the international joint program, CarboEastAsia. The program was conducted by three networks in Asia, ChinaFLUX, JapanFlux, and KoFlux, to quantify, synthesize, and understand the carbon budget of the eastern part of Asia. An intercomparison was conducted for NEE estimated by three gap-filling procedures adopted by ChinaFLUX, JapanFlux, and KoFlux to test the range of uncertainty in the estimation of NEE. The overall comparison indicated good agreement among the procedures in the seasonal patterns of NEE, although a bias was observed in dormant seasons depending on the different criteria of data screening. Based on the gap-filled datasets, the magnitude and seasonality of the carbon budget were compared among various biome types, phenology, and stress conditions throughout Asia. The annual values of gross primary production and ecosystem respiration were almost proportional to the annual air temperature. Forest management, including clear-cutting, plantation, and artificial drainage, was significant and obviously affected the annual carbon uptake within the forests. Agricultural management resulted in notable seasonal patterns in the crop sites. The dataset obtained from a variety of biome types would be an essential source of knowledge for ecosystem science as well as a valuable validation dataset for modeling and remote sensing to upscale the carbon budget estimations in Asia.     

Effects of Acid Deposition on Urushi Lacquer in East Asia

Urushi (Japanese lacquer) plates were exposed to indoor air in 7 cities in East Asia from summer 1995 through winter 1997. The plates, collected every 3 months, were optically observed by using a gloss meter, a digital microscope and a microscopic infrared spectrophotometer for evaluation of a seasonal impact by acidic air pollutants. The gloss losses were high on the urushi surface in autumn, when fog frequently appeared. Numerous fine spots were observed in 0.2–0.3 mm in diameter on the surfaces of the plates exposed at Chongqing, China, Taejon, Korea and Nara, Japan, where dense fog occurred. The spots were dark and opaque at Chongqing, where heavy air pollution was observed. A dark and opaque core was observed in each semitransparent spot at Taejon and Nara, while no spots were observed at Kyoto and Ishikawa, where fog often appeared but air pollution was at a low level. Carbonyl group, identified by microscopic infrared spectrometry, was found in the spots on the urushi surfaces. The carbonyl group may be formed by oxidation of a side-chain in urushiol (a major component of urushi sap, alkyl phenol). Urushi lacquer may be damaged by high concentration of sulfate anion, included in acid fog.     

Slow earthquakes linked along dip in the Nankai subduction zone

We identified a strong temporal correlation between three distinct types of slow earthquakes distributed over 100 kilometers along the dip of the subducting oceanic plate at the western margin of the Nankai megathrust rupture zone, southwest Japan. In 2003 and 2010, shallow very-low-frequency earthquakes near the Nankai trough as well as nonvolcanic tremor at depths of 30 to 40 kilometers were triggered by the acceleration of a long-term slow slip event in between. This correlation suggests that the slow slip might extend along-dip between the source areas of deeper and shallower slow earthquakes and thus could modulate the stress buildup on the adjacent megathrust rupture zone.     

Study of the role of the carbohydrate and protein moieties of soy soluble polysaccharides in their emulsifying properties

The objective of this work was to investigate the role played by the protein fraction of soy soluble polysaccharide (SSPS) during its adsorption at oil/water interfaces. SSPS was separated in a high (HMF; 310 kDa) and low (LMF; 20 kDa) molecular weight fraction by gel filtration. SSPS/HMF consisted of 91.6% carbohydrate and 2.2% protein and showed better emulsifying properties than those of the whole SSPS, whereas SSPS/LMF seemed to affect negatively the adsorption behavior of SSPS. SDS-PAGE of the protein fraction obtained from SSPS/HMF showed a molecular mass of 50 kDa, was composed predominantly of proline (23.1%) and glutamic acid (15.2%), and still contained 8.8% of neutral sugar and 5.3% of uronic acid. Results indicated that not all of the protein material present in SSPS contributes to SSPS functionality but that only the material associated with HMF aids in the adsorption of SSPS onto oil/water interfaces.     

Structural characterization and biological activities of an exopolysaccharide kefiran produced by Lactobacillus kefiranofaciens WT-2B(T)

Lactobacillus kefiranofaciens, isolated from kefir grains, produces an extracellular polysaccharide when cultured, not only in PYG10 medium but also in a liquid medium containing a rice hydrolysate that had been previously degraded by treatment with a glucoamylase. The maximum yield of the polysaccharide, using the rice hydrolysate as the medium, was 2.5 g/L after a 7-day culture period at pH 5.0 and 33 degrees C. Compositional analysis, methylation analysis, specific rotation, and (1)H and (13)C NMR spectroscopy revealed that the structures of polysaccharides obtained from these two different culture media are essentially identical. The polysaccharide is composed of a hexasaccharide repeating unit and, thus, is known as kefiran. The weight-average molecular weight and the z-average radius of gyration of a sample, purified from the rice hydrolysate medium, were determined to be 7.6 x 10(5) g/mol and 39.9 nm, respectively, by gel permeation chromatography equipped with a multiangle laser-light-scattering photometer. Changes in blood pressure and serum components were examined in SHRSP/Hos rats, using doses of 100 and 300 mg of kefiran/kg of rat. A suppression in the increase in blood pressure was observed in these rats after 30 days. This activity is discussed in terms of the concentration of serum components of the rat, with emphasis on lipid components such as cholesterols, triglycerides, and free fatty acids.