Development of a Medium-term Animal Model Using gpt Delta Rats to Evaluate Chemical Carcinogenicity and Genotoxicity

In this study, the potential for development of an animal model (GPG46) capable of rapidly detecting chemical carcinogenicity and the underlying mechanisms of action were examined in gpt delta rats using a reporter gene assay to detect mutations and a medium-term rat liver bioassay to detect tumor promotion. The tentative protocol for the GPG46 model was developed based on the results of dose-response exposure to diethylnitrosamine (DEN) and treatment with phenobarbital over time following DEN administration. Briefly, gpt delta rats were exposed to various chemicals for 4 weeks, followed by a partial hepatectomy (PH) to collect samples for an in vivo mutation assay. The mutant frequencies (MFs) of the reporter genes were examined as an indication of tumor initiation. A single intraperitoneal (ip) injection of 10 mg/kg DEN was administered to rats 18 h after the PH to initiate hepatocytes. Tumor-promoting activity was evaluated based on the development of glutathione S-transferase placental form (GST-P)-positive foci at week 10. The genotoxic carcinogens 2-acetylaminofluorene (2-AAF), 2-amino-3-methylimidazo [4,5-f] quinolone (IQ) and safrole (SF), the non-genotoxic carcinogens piperonyl butoxide (PBO) and phenytoin (PHE), the non-carcinogen acetaminophen (APAP) and the genotoxic non-hepatocarcinogen aristolochic acid (AA) were tested to validate the GPG46 model. The validation results indicate that the GPG46 model could be a powerful tool in understanding chemical carcinogenesis and provide valuable information regarding human risk hazards.     

Blockade of bovine PD-1 increases T cell function and inhibits bovine leukemia virus expression in B cells in vitro

Programmed death-1 (PD-1) is a known immunoinhibitory receptor that contributes to immune evasion of various tumor cells and pathogens causing chronic infection, such as bovine leukemia virus (BLV) infection. First, in this study, to establish a method for the expression and functional analysis of bovine PD-1, hybridomas producing monoclonal antibodies (mAb) specific for bovine PD-1 were established. Treatment with these anti-PD-1 mAb enhanced interferon-gamma (IFN-γ) production of bovine peripheral blood mononuclear cells (PBMC). Next, to examine whether PD-1 blockade by anti-PD-1 mAb could upregulate the immune reaction during chronic infection, the expression and functional analysis of PD-1 in PBMC isolated from BLV-infected cattle with or without lymphoma were performed using anti-PD-1 mAb. The frequencies of both PD-1⁺ CD4⁺ T cells in blood and lymph node and PD-1⁺ CD8⁺ T cells in lymph node were higher in BLV-infected cattle with lymphoma than those without lymphoma or control uninfected cattle. PD-1 blockade enhanced IFN-γ production and proliferation and reduced BLV-gp51 expression and B-cell activation in PBMC from BLV-infected cattle in response to BLV-gp51 peptide mixture. These data show that anti-bovine PD-1 mAb could provide a new therapy to control BLV infection via upregulation of immune response.     

Effect of wood species on thermal flow behavior and physical properties of thermoplastic moldings

The thermoplastic flow behavior of cedar flour steamed at different temperatures in the range of 160–220°C was measured using a rheometer and compared with that of beech flour. The temperature at which the cedar flour starts to flow was approximately 70°C higher than that of beech flour, and the cedar flour exhibited low flowability. Furthermore, thermoplastic moldings were prepared from cedar and beech flours, and their physical properties were examined. Similar to the case of beech, a resin-like molding with a density of approximately 1.45?g/cm³ was obtained from the cedar flour steamed at 180°C or higher, and it was revealed by SEM observation that in these moldings wood flour particles adhere to each other. The specific bending strength was maximum for the moldings obtained from 180°C-steamed flour for both types of wood.     

A refined hydro-environmental watershed model with field-plot-scale resolution

A distributed hydro-environmental model is developed that achieves detailed analysis of the movement of water at a field-plot-scale resolution in a mesoscale watershed including lowland areas where, especially for agricultures, it is an essential need to get rid of redundant groundwater by drainage facilities such as rivers, canals and/or underdrains. For this, the problem geometry is meshed with unstructured cells of triangular shape. Profile of a column cell is zoned into two: surface zone and groundwater zone in which water movement is represented by combined tank and soil moisture sub-models, and well-defined two-dimensional unconfined shallow groundwater flow sub-model, respectively. The top-two sub-models serve to evaluate evapotranspiration, infiltration, soil water content, lateral surface water flow, and vertical percolation. The vertical percolation so evaluated is given as longitudinal recharge to the bottom sub-model for computing groundwater flow. Surface water–groundwater interactions through beds and stream-banks of perennial and ephemeral canals are considered by treating the canal courses as internal boundaries in the groundwater flow model. The finite volume method (FVM) that allows of unstructured mesh and produces conservative solutions is employed for groundwater flow computation. The model developed is applied to an actual watershed which includes a low-lying paddy area to quantify the hydrological impact of land-use management practices over a period of 29 years in which the farmland consolidation project was implemented and part of the paddy fields were converted to upland crop fields and housing lands. From the results obtained, it is concluded that the model presently developed lends itself to water—as well as land-use management practices.     

A hydro-environmental watershed model improved in canal-aquifer water exchange process

Long-term simulation using the distributed hydro-environmental watershed model is efficacious for assessing irrigation impacts on hydrological cycle in detail and for implementing watershed management successfully. In this article, the previously developed hydro-environmental watershed model (HEWM-1) is improved in the water exchange process caused by surface water-groundwater interaction via drainage canals and/or underdrains. The time-varying stream flow in canals is described by the complete one-dimensional shallow water equations in a newly introduced submodel, the open channel flow submodel. This submodel coordinates with the other submodels: the tank, soil moisture and groundwater flow submodels which are interlinked in a cascade manner. The improved model (HEWM-2) is applied to an agricultural watershed covering an area from an alluvial fan onto a nearly level alluvial plain, to be validated. The simulation by HEWM-2 is informative for identifying whether any drainage canal is gaining or losing water in relation to groundwater level. It could thus provide useful information for conserving a complex network of drainage canals which also functions as a passage for aquatic animals like fishes.     

Spontaneous Erythroid Leukemia in a 7-Week-Old Crl:CD (SD) Rat

A young male Crl:CD (SD) rat with erythroid leukemia that presented with emaciation, abdominal distension and a pale visible mucosal membrane was euthanized at 7 weeks of age. At necropsy, enlargement of liver, spleen and pancreatic lymph node was noted. Analysis of blood smear samples revealed many mono- or binucleated erythroblasts that had PAS-positive vacuoles in the cytoplasm. Histopathologically, neoplastic proliferation of atypical cells was observed in the hepatic sinusoids, splenic red pulp, bone marrow, pancreatic lymph node, kidney and lung. Neoplastic cells showed a round to spindle shape, and some neoplastic cells had deeply stained small nuclei and small cytoplasms and resembled erythroblasts. Immunohistochemically, many neoplastic cells were positive for hemoglobin. To our knowledge, this is the first report of erythroid leukemia in a rat of this age. The observed features were similar to those of pure erythroid leukemia in humans.     

Novel Wx alleles induced by chemical mutagens in rice,Oryza sativa L.

From mutant pools of two Taiwanese elite japonica cultivars, Tainung 67 and Taikeng 8, we identified 13 mutant lines possessing opaque endosperm with relatively low amylose contents (AC) ranging from 1.5% to 7.1%. Because of different AC, paste viscosities of these 13 mutant lines differed, as revealed by palatability and physicochemical properties. The mutated gene conferring opaque endosperm was isolated from the F₂ population of one mutant line, WY1× indica cv. ‘Taichung Sen 17’, by positional cloning, revealing a G3018→A3018 substitution at exon 9 of Waxy leading to a non‐synonymous mutation from alanine to valine. Two additional alleles were identified from the other 12 mutant lines, for which single‐nucleotide substitutions G2708 → A2708 and G3029 → A3029 occurred in exons 8 and 9, leading to non‐synonymous mutations from arginine to histidine and glutamic acid to lysine, respectively. The three novel wx alleles had different effects on grain quality, specifically on eating and cooking quality, and could be applied in rice breeding programmes to develop new low AC varieties by marker‐assisted selection.     

Structural identification of anthocyanins and analysis of concentrations during growth and flowering in buckwheat (Fagopyrum esculentum Moench) petals

The anthocyanin profiles and variety/breeding-line differences of anthocyanin concentrations in petals of common buckwheat flowers have been studied. Four anthocyanins, cyanidin 3-O-glucoside, cyanidin 3-O-rutinoside, cyanidin 3-O-rhamnoside, and cyanidin 3-O-galactosyl-rhamnoside were isolated from the petals of common buckwheat (Fagopyrum esculentum Moench), separated using high performance liquid chromatography and identified using reversed-phase liquid chromatography-electrospray ionization-tandem mass spectrometry techniques. In every variety/breeding line tested, cyanidin 3-O-rutinoside was detected as the major anthocyanin and the next is cyanidin 3-O-glucoside whereas cyanidin 3-O-rhamnoside and cyanidin 3-O-galactosyl-rhamnoside were trace or not detectable in white and pink flowered buckwheat. Of all the varieties/breeding lines tested, Gan-Chao, a Chinese variety, contained the highest amount of anthocyanins. The largest part of cyanidin moiety was presented as a proanthocyanidin form (PAs-Cy). Anthocyanins and PAs-Cy in petals were increased along with increase of flower development stages. Therefore, fully developed petals of red flowered buckwheat, especially Gan-Chao, are promising as a new anthocyanin-rich material for food processing.     

Contribution of different proton sources to pedogenetic soil acidification in forested ecosystems in Japan

The contribution of different proton sources to pedogenetic soil acidification was evaluated for three Japanese forest soils, i.e. ando soil, podzolic soil and brown forest soil in relation to the respective soil forming processes. Soil acidification rate and net proton generation were quantified based on the theory of proton budget for the respective soil horizon compartments (mainly the O, A and B horizons) by measuring fluxes of solutes entering and leaving the soil horizon compartment and vegetation uptake. Protons were produced by the dissociation of organic acids and nitrification in the O horizon and then consumed by adsorption and decomposition of organic acids and nitrate uptake by vegetation in deeper soil horizons at all plots. Excess uptake of cation over anion by vegetation was highest among proton sources in the whole soil compartment at all plots. Pedogenetic soil acidification was considered to include cation leaching from surface soil horizons due to proton generation by the dissociation of organic acids and nitrification and subsequent cation excess accumulation in wood in the growth stage of forests. In ando soil, andosolization resulted from the low contribution of net proton generation by the dissociation of organic acids as well as a lower soil acidification rate and complete acid neutralization. Dissolved organic carbon (DOC) fluxes in ando soil were lower than those in podzolic soil and brown forest soil due to high adsorption capacity of amorphous materials. In podzolic soil, podzolization resulted from intensive acidification in the O horizon, which derived from net proton generation by the dissociation of organic acids and nitrification as well as cation excess uptake by vegetation due to concentrated fine root biomass in the O horizon, and subsequent high proton efflux to subsoil. The high fluxes of DOC and Al leached from surface soil horizons were considered to contribute to eluviation of Al from surface soil and illuviation in subsoil in podzolic soil. In brown forest soil, brunification resulted from a lower DOC flux from the O horizon due to high decomposition and adsorption by oxides, where podzolization was weakened by high acid neutralization. Thus, the three representative processes involved in the pedogenesis of Japanese forest soils were well characterized by quantification of the respective proton-generating and consuming processes in each soil horizon.     

Sequencing of canine 5-hydroxytriptamine receptor (5-HTR) 1B, 2A, 2C genes and identification of polymorphisms in the 5-HTR1B gene

Polymorphisms of human genes encoding 5-hydroxytriptamine (serotonin) receptors (5-HTRs) are thought to be associated with psychiatric disorders and behavioral traits. In the present study, we searched for corresponding polymorphisms in the dog and compared allelic frequencies for the canine 5-HTR1B, 5-HTR2A, and 5-HTR2C genes among five canine breeds. The canine genes consisted of the following: 5-HTR1B, 1170 bp; 5-HTR2A, 1413 bp; and 5-HTR2C, 1377 bp. All of these genes were highly homologous with the human genes. We found six single nucleotide polymorphisms (SNPs) in the 5-HTR1B gene (G57A, A157C, G246A, C660G, T955C, and G1146C). Genotyping of the respective SNPs revealed that there were inter-breed variations in the genotypes and allelic frequencies for four out of the six identified SNPs, suggesting that further analyses of the polymorphisms of the 5-HTR1B gene would be useful in order to gain an understanding of the genetic background underlying the diversified behavioral traits among canine species.