Effect of a Large Dose of Di (2-ethylhexyl) phthalate (DEHP) on Hepatic Peroxisome in Cynomolgus Monkeys (Macaca Fascicularis)

To elucidate the effect of a large dose of di (2-ethylhexyl) phthalate (DEHP), a plasticizer and peroxisome proliferator-activated receptor-α (PPARα) agonist, on hepatic peroxisomes, we orally administered 1,000 mg/kg/day, once daily, to 3 male and 4 female cynomolgus monkeys for 28 days consecutively. Light-microscopic and electron microscopic examinations of the liver were carried out in conjunction with measurement of the hepatic fatty acid β-oxidation system (FAOS), carnitine acetyltransferase (CAT) and carnitine palmitoyltransferase (CPT) activities, which are peroxisomal and/or mitochondrial enzyme activities. Electron microscopically, enlargement of the mitochondria was observed with lamellar orientation of the cristae along the major axis. Although the number of peroxisomes showed a tendency to increase when compared with those in a biopsied specimen before treatment, no abnormality in morphology was observed. A slight increase in CPT activity was noted at termination. No changes were noted in hepatic FAOS or CAT activity. In conclusion, although repeated oral treatment of cynomolgus monkeys with a large dose of DEHP induced a subtle increase in the numbers of peroxisomes with slight enlargements of the mitochondria, this low-sensitivity response to peroxisome proliferators in cynomolgus monkeys was considered to be closer to the response in humans than that in rodents.     

Epidemiological survey of Border disease virus among sheep from northern districts of Japan

The first epidemiological survey of Border disease virus (BDV) was undertaken in small ruminants in Japan. Ovine sera, collected from the northern prefectures of Hokkaido, Aomori and Iwate, were examined for the presence of antibodies against BDV using the neutralization peroxidase-linked antibody test. Twenty-nine (17.6%) of one hundred and sixty-five samples were seropositive for BDV. Results were specific, excluding cross-reactions with bovine viral diarrhea virus (BVDV). Only one sample (0.6%) was positive for BVDV, and was negative for BDV. Despite serological evidence of virus circulation, there have been no clinical cases of border disease in sheep in Japan. Although no diagnostic measures were performed, the infection did not appear to be associated with a reduction in ewe fertility nor with lamb mortality.     

Glomerular lipidosis accompanied by renal tubular oxalosis in wild and laboratory-reared Japanese rock ptarmigans (Lagopus mutus japonicus)

Glomerular lipidosis is a disease characterized by lipid accumulation in mesangial cells but that has not been fully investigated in avian species. We examined four wild and two laboratory-reared Japanese rock ptarmigans (Lagopus mutus japonicus)--an endangered avian species--presenting vacuolar deposits in the glomeruli. All cases had vacuolar deposits in the glomeruli. In the wild cases, fewer than 30% of all glomeruli were affected, compared with more than 90% in the laboratory-reared cases. In the wild cases, most deposits were mild and restricted to the mesangial areas of glomeruli. In the laboratory-reared cases, nearly all of the deposits covered entire glomeruli. Electron microscopy of mild deposits revealed vacuoles in the cytoplasm of mesangial cells. These vacuoles were positive for Sudan III, Sudan black B, oil red O, Nile blue, periodic acid-Schiff, Schultz test, and digitonin stain and were negative for performaric acid-Schiff stains. Based on these results, we diagnosed the glomerular lesion as glomerular lipidosis caused by uptake of low-density lipoprotein in mesangial cells. Except for one wild case, all cases exhibited renal tubular oxalosis. The severity of tubular oxalosis tended to be related to the severity of glomerular lipidosis: In cases of mild glomerular lipidosis, tubular oxalosis was also mild or absent. We therefore diagnosed the primary lesion as glomerular lipidosis accompanied by tubular oxalosis. The four wild cases came from different zones and therefore had no opportunities to interbreed and no common relatives. We believe these data support the hypothesis that glomerular lipidosis is a disease of the general population ofJapanese rock ptarmigans. This is the first report of glomerular lipidosis accompanied by renal tubular oxalosis in an avian species.     

Classification of the genus Bacillus based on MALDI-TOF MS analysis of ribosomal proteins coded in S10 and spc operons

A rapid bacterial identification method by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) using ribosomal proteins coded in S10 and spc operons as biomarkers, named the S10-GERMS (the S10-spc-alpha operon gene encoded ribosomal protein mass spectrum) method, was applied for the genus Bacillus a Gram-positive bacterium. The S10-GERMS method could successfully distinguish the difference between B. subtilis subsp. subtilis NBRC 13719(T) and B. subtilis subsp. spizizenii NBRC 101239(T) because of the mass difference of 2 ribosomal subunit proteins, despite the difference of only 2 bases in the 16S rRNA gene between them. The 8 selected reliable and reproducible ribosomal subunit proteins without disturbance of S/N level on MALDI-TOF MS analysis, S10, S14, S19, L18, L22, L24, L29, and L30, coded in S10 and spc operons were significantly useful biomarkers for rapid bacterial classification at species and strain levels by the S10-GERMS method of genus Bacillus strains without purification of ribosomal proteins.     

How different or similar are nematode communities between a paddy and an upland rice fields across a flooding-drainage cycle?

The paddy field is being recognised as a biodiversity hotspot fostering a variety of organisms. However, there are few studies on the ecology of paddy field nematodes. We characterised nematode communities in rice paddy fields by comparing them with upland fields of rice or soybean. We examined nematode communities of the top (0-15 mm) and second (15-50 mm) soil layers before flooding (March or April), during flooding (June or July) and during the draining period (October) 2007-2009. We found that the nematode community in the paddy was different than that in the upland fields during all periods. Rhabdolaimus, Tobrilus, Mesodorylaims and Monhysteridae characterised the top of the paddy and Hirschmanniella characterised the second layer of the paddy. Total nematode density was generally lower in the paddy than in the upland field. However, the density in the paddy top layer increased with time from the flooding period to the draining period, during which time it was about the same as (or even greater than) the peak density in the upland fields. The density in the second layer of the paddy remained lower than that in the top layer of the paddy throughout the time course. Community diversity values were generally greater in the paddy top layer than in the paddy second layer across the six sampling periods, but periodic differences between the paddy and upland fields or between soil layers were not significant. During the flooding period, the F/(F + B) (13-37) and Enrichment Index (17-38) values were lower in the paddy than in the upland fields (32-47, 37-74, respectively) to reflect that bacteria dominate over fungi with slow decomposition due to anaerobic conditions in the flooded paddy field. In addition, particularly in the top layer, the Maturity (2.0-2.4) and Structure Index (23-72) values were greater in the paddy than in the upland fields (1.7-2.1, 9-15, respectively), indicating a well-developed ecosystem under water. These unique nematode communities persisted during the draining period, but there was a rapid increase in opportunistic bacterivores, which increased the EI values. We suggest that bactivorous nematodes in the families Cephalobidae and Chronogasteridae, herbivores in the genus Hirschmanniella, and fungivores in the genus Filenchus may be specific to paddy field soil rather than to pond and lake sediments.     

A chlorite-smectite intergrade in soil derived from pyroxene andesite altered hydrothermally in Mt. Kunimidake,Fukui, Central Japan

The interlayered hydroxy-aluminum of a chlorite-smectite intergrade in a dark-red soil derived from a pyroxene andesite altered by hydrothermal action was extracted easily by boiling treatments using both sodium-citrate and ammonium-fluoride solutions. Moreover, this interlayered aluminum was released partially by using a dithionite-citrate-bicarbonate system and an acid ammonium-oxalate solution which were prepared originally for removing the coexistent sesquioxides from soils.     

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.     

Cloning of the phloem-specific small heat-shock protein from leaves of rice plants

The N-terminal amino-acid sequence of a major rice phloem-sap protein, designated as RPP23, was determined. The complete amino-acid sequence of RPP23 was deduced from the corresponding rice EST- clone and contained an extra 46 amino acids at the N-terminus, that was apparently cleaved off to form mature RPP23 in sieve tubes. RPP23 shared a similarity to plant small heat-shock proteins (smHSPs), though the N-terminal region of RPP23 was distinct from that of known smHSPs. Immunocytological analyses using leaf sections showed that RPP23 was located only in the phloem regions of leaves, and was present in non-stressed plants. In mature leaves, stronger immunocytological signals were detected in sieve elements than in companion cells.