Isolation and characterization of macrophages from a mixed primary culture of bovine liver cells

Previously, we developed a simple and efficient method to isolate liver macrophages from a mixed primary culture of adult rat liver cells. To extend the applicability of this method, we isolated macrophages from mixed primary cultures of bovine liver cells. Macrophage cells proliferated on the cell sheet of mixed bovine liver cells after 8-16d of culture. These cells were detached by shaking of the culture flasks. Subsequent transfer and brief incubation in plastic dishes resulted in selective adhesion of macrophages. After rinses with PBS, attached macrophages were harvested. More than 10(6) cells could be harvested from the culture flask at intervals of 2-3d for more than three weeks. The isolated cells were strongly positive for bovine macrophage markers, such as CD68, CD172a and Iba-1. These cells exhibited functional properties of macrophages, including active phagocytosis of polystyrene microbeads, proliferative response to recombinant bovine granulocyte-macrophage colony-stimulating factor, upregulation of specific inflammatory cytokine genes upon stimulation with lipopolysaccharide, and formation of multinucleated giant cells. The shaking and attachment method provides a simple and efficient alternative to obtain bovine liver macrophages without requiring complex equipment or specialized technical skills.     

Root-shoot interaction as a limiting factor of biomass productivity in new tropical rice lines

The objective of the present study was to identify the limiting factors in biomass productivity of new tropical rice lines (panicle weight type) by analyzing the relationship between root and shoot activity. Five field experiments using three new lines, IR65598-112-2, IR65600-42-5-2, and IR68544-29-2-1-3-1-2, and one of the highest-yielding indica rice varieties (panicle number type), IR72, were conducted at the International Rice Research Institute from 1997 to 2002. Specific absorption rate of N (SARN , mg N g^-1 root dry weight d^-1), specific stem sap exudation rate (sap exudation rate, g exudate g^-1 root dry weight 12 h^-1) and specific root respiratory rate (root respiratory rate, μmol O₂ g^-1 root dry weight h^-1) were determined as indices of root activity. Relative growth rate (RGR, mg g^-1 d^-1) was used as an index of shoot activity. Compared with IR72, the new lines showed lower RGR and SAR_N values during the early growth stages. In contrast, during the late growth stages, these activities were higher in the new lines than in IR72. The SAR_B and sap exudation rate showed a linear correlation with RGR at successive growth stages, regardless of the genotypes and growth conditions. These findings indicated that active root-shoot interaction was maintained throughout the growth period in high-yielding tropical rice, including the new lines and IR72. Therefore, it was considered that SAR_N and the sap exudation rate are useful indices of root activity regulating RGR in the new lines. However, the root respiratory rate could not be used for selection, because the relationship between RGR and the root respiratory rate changed with the growth conditions. Our findings support the hypothesis that root and shoot activities during maturation are important factors affecting the productivity of the new lines, which have not yet been able to attain the targeted yield.     

Varietal difference in radiocesium uptake and transfer from radiocesium deposited soils in the genus Amaranthus

Abstract

Within Amaranthaceae, 33 different varieties, including local varieties from Japan, were grown in 2012 in a field in the town of Iino in the Fukushima prefecture, which is located approximately 51 km north of Tokyo Electric Power Company, Fukushima Daiichi Nuclear Power Plant (FDNPP). The contamination level of the soil was 2770 ± 140 Bq kg^?1 dry weight (¹³⁴Cesium (Cs) + ¹³⁷Cs, average ± SE), and the field was also cultivated in 2011. There was a significant varietal difference in the dry weight production, radiocesium accumulation and transfer factor (TF) of radiocesium from the soil to the plant. The ratio of the lowest TF to the highest TF was approximately 3. Because the ratio of ¹³⁷Cs to ¹³³Cs was significantly positive, radiocesium seems to be absorbed in a manner similar to that of ¹³³Cs. It is suggested that the varietal difference in the behavior of radiocesium uptake mainly depends on its genetic background rather than on environmental factors.     

Mechanisms of Aluminum Tolerance in Phosphoenolpyruvate Carboxylase Transgenic Rice

Transgenic rice, in which C4-phosphoenolpyruvate carboxylase (PEPC) is overexpressed, was grown in hydroponic culture to evaluate its tolerance for high aluminum (Al) content in relation to organic acid synthesis and exudation into the rhizosphere. With regard to root growth in short-term experiments, PEPC-transgenic rice was more tolerant of Al than the wild type. At increased Al levels in the root medium, root tips of the transgenic rice accumulated less Al than those of the wild type. Aluminum-induced oxalate exudation from roots occurred at increased rates in the transgenic line. Overexpression of C4-PEPC drastically increased PEPC activity in leaves of the transgenic rice, causing higher organic acid concentrations in leaves and roots. The increased organic acid concentrations seemed to cause higher exudation rates of oxalate from roots, leading to a detoxification of Al in the root apoplast and rhizosphere.     

Water penetration into rice grains during soaking observed by gradient echo magnetic resonance imaging

The water penetration into a rice grain during soaking was monitored by three-dimensional gradient echo magnetic resonance imaging, which has the advantage of a short measurement time and a sufficient sensitivity for low-moisture samples. The water penetration route was compared between the milled and brown rice grains of two japonica cultivars. In normal milled rice grains of Koshihikari, water first penetrated the ventral side surface and the embryo attachment site of the endosperm, then migrated along the central line and transverse cracks, and finally diffused to all parts of the endosperm. In milled rice grains of Yamadanishiki with a white core, water quickly infiltrated into the cracks or chalky parts on the dorsiventral line and then diffused to the lateral side. In brown rice grains of both cultivars water penetrated extremely slowly and did not infiltrate directly into transverse cracks or the white core due to inhibition by the pericarp and seed coat. The central part of every grain allowed more water to penetrate. The route, pattern and speed of water penetration were determined by the morphological structure, crack formation and hardness distribution associated with the packing of the starch granules in the grain.     

Effect of boiled barley-rice-feeding in hypercholesterolemic and normolipemic subjects

Barley contains approximately 10% dietary fiber and is easily cooked with rice, the dominant cereal in Japan, to increase the intake of dietary fiber. This research involved three experiments to examine the influence of barley on blood lipids in human subjects. All subjects received a boiled barley-rice (50/50 w/w mix) supplement two times per day in place of rice for 2 or 4 weeks. In the normolipemic subjects, serum lipids were unaffected by the ingestion of barley for 4 weeks. In twenty hypercholesterolemic men aged 41 ± 5 years, the ingestion of barley was associated with a significant fall in serum total cholesterol, LDL-cholesterol, phospholipids and LDL and VLDL-lipoproteins. In seven mildly hypercholesterolemic women aged 56±7 years, a significant improvement of serum lipid profiles was observed. The present study suggests the possibility that the ingestion of barley-rice could lower serum lipids in hypercholesterolemic subjects.     

Texture measurement of cabbages using an acoustical vibration method

Textures of six cabbage cultivars were quantified using an acoustical vibration technique. A sample of four outer leaves of a cabbage was penetrated using a probe. The acoustical vibration signals were measured during penetration using a piezoelectric sensor. A new texture index (TI), the “energy density”, was introduced, which was determined by the integration of squared amplitudes of texture signals multiplied by a factor of a frequency band. This TI enabled evaluation of acoustical signals in the high-frequency region (>1000 Hz) more sensitively than the previously used index (“amplitude density”), which was determined by the integration of texture signal amplitudes. Significant differences in TI among the cultivars were obtained by using ANOVA, especially between a spring and a winter cabbage. We also examined cabbages that had been stored under 4 °C for 10 or 19 d. Most TI readings increased after the storage. These results provide useful information related to the shelf-life of cabbages.     

Molecular and biological characterization of <Emphasis Type="Italic">Chrysanthemum stem necrosis virus</Emphasis> isolates from distinct regions in Japan

Three isolates of Chrysanthemum stem necrosis virus (CSNV) were obtained from chrysanthemum plants in distinct regions of Japan in 2006 and 2007. All the original host plants showed severe necrotic symptoms on the leaves and stems. Amino acid sequence data of the nucleocapsid protein genes of the three isolates (CbCh07A, TcCh07A, and GnCh07S) showed high identities with those of two other CSNV isolates, HiCh06A L1 from Japan and Chry1 from Brazil. Furthermore, for the first time the complete nucleotide sequence of the S RNA was determined for CSNV (isolate HiCh06A). In phylogenetic analysis based on the non-structural protein genes from the genus Tospovirus, HiCh06A L1 was placed in the same genetic group as Tomato spotted wilt virus (TSWV) and Impatiens necrotic spot virus. Host range examination for isolates HiCh06A L1 and CbCh07A showed that green pepper (cv. ‘Kyoyutaka’, ‘Saitamawase’, ‘Tosakatsura’, ‘L3 sarara’ and ‘L3 miogi’) and tomato (cv. ‘Sekaiichitomato’) were systemically susceptible hosts, whereas TSWV-resistant Solanaceae species, Capsicum chinense, Lycopersicon peruvianum and a TSWV-resistant cultivar of green pepper (cv. TSR miogi), were resistant.     

Molecular diversity of binucleate <Emphasis Type="Italic">Rhizoctonia</Emphasis> AG-A in China

AG-A belongs to the binucleate Rhizoctonia (BNR) anastomosis group (AG) of the Ceratobasidium teleomorph, which parasitizes the roots of many plant species. Ninety nine isolate species of AG-A were obtained from Tibet, Sichuan, and Yunnan Province in China. All isolates were divided into three types based on their cultural characteristics. Type I: abundant aerial mycelia, dense hyphae, loose sclerotia; Type II: abundant aerial mycelia, no sclerotia. Type III: sparse aerial mycelium and no sclerotia. All of the isolates infected the seedlings of Chinese mustard and Chinese cabbage, causing the formation of lesions on the stem and a brown discoloration of the roots. Sequence analysis of the 5.8S rDNA-ITS showed a similarity of 98–100% among the isolates. Inter Simple Sequence Repeat (ISSR) was used to detect genetic variation in binucleate Rhizoctonia spp. Forty two AG-A isolates were amplified using 15 random primers. From a total of 164 bands, 144 bands (87.8%) were polymorphic in the 42 tested isolates. A dendrogram showing genetic relationships between the isolates was constructed using unweighted pair-group averages based on genetic distances. According to the dendrogram, the 42 tested isolates could be aligned into three clusters with a genetic similarity coefficient of 0.29, the first clusters including 27 isolates with III of culture characteristics on PDA; the second clusters included eight isolates with I of cultural characteristics on PDA; the third cluster included seven isolates with II of cultural characteristics on PDA. The results of ISSR analysis showed an association between the hosts of these isolates. Our results showed that ISSR analysis can reveal more molecular variation among isolates of AG-A than sequence analysis using the 5.8S rDNA-ITS.