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.     

Structural reappraisal of corticatic acids,biologically active linear polyacetylenes,from a marine sponge of the genus <Emphasis Type="Italic">Petrosia</Emphasis>

Structural reinvestigation of corticatic acid A from a marine sponge, Petrosia sp., resulted in the revision of its structure, which conformed to the structural features of the biosynthetically related linear acetylenes isolated from Petrosia sp. In the process of isolating an authentic corticatic acid A, we isolated three new congeners and determined their structures. We also suggested that the structures of other corticatic acids need revision.     

Cryopreservation of male and female gonial cells by vitrification in the critically endangered cyprinid honmoroko <Emphasis Type="Italic">Gnathopogon caerulescens</Emphasis>

We investigated the feasibility of cryopreservation of spermatogonia and oogonia in the critically endangered cyprinid honmoroko Gnathopogon caerulescens using slow-cooling (freezing) and rapid-cooling (vitrification) methods. Initially, we examined the testicular cell toxicities and glass-forming properties of the five cryoprotectants: ethylene glycol (EG), glycerol (GC), dimethyl sulfoxide (DMSO), propylene glycol (PG), and 1,3-butylene glycol (BG), and we determined cryoprotectant concentrations that are suitable for freezing and vitrification solutions, respectively. Subsequently, we prepared the freezing solutions of EG, GC, DMSO, PG, and BG at 3, 2, 3, 2, and 2 M and vitrification solutions at 7, 6, 5, 5, and 4 M, respectively. Following the cryopreservation of the testicular cells mainly containing early-stage spermatogenic cells (e.g., spermatogonia and primary spermatocytes), cells were cultured for 7 days and immunochemically stained against germ cell marker protein Vasa. Areas occupied by Vasa-positive cells indicated that vitrification led to better survival of germ cells than the freezing method, and the best result was obtained with 5 M PG, about 50% recovery of germ cells following vitrification. In the case of ovarian cells containing oogonia and stage I, II, and IIIa oocytes, vitrification with 5 M DMSO resulted the best survival of oogonia, with equivalent cell numbers to those cultured without vitrification. The present data suggest that male and female gonial cells of the endangered species G. caerulescens can be efficiently cryopreserved using suitable cryoprotectants for spermatogonia and oogonia, respectively.     

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.     

Antigenic and genetic analyses of H5 influenza viruses isolated from ducks in Asia.

The hemagglutinin (HA) of six H5 influenza virus strains isolated from ducks in Japan and China in 1976 to 1996 were analyzed antigenically and genetically. Antigenic analysis using a panel of monoclonal antibodies revealed that the HA of H5 influenza viruses isolated from ducks are antigenically closely related to each other. Phylogenetic analysis indicates that the isolates from ducks in Hokkaido were derived from an ancestor common with the highly pathogenic isolates from chickens and humans in Hong Kong in 1997.     

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.     

High incidence of the gullet worm, Gongylonema pulchrum, in a squirrel monkey colony in a zoological garden in Japan

Histological examination revealed the gullet worm (Gongylonema pulchrum) embedded in the lingual mucosa of two of four dead Bolivian squirrel monkeys (Saimiri boliviensis) from a zoological garden in Kyushu, Japan. The zoo had kept the monkeys as a colony of approximately 80-100 individuals in a moated, open ground since 1981. To assess the incidence of G. pulchrum infection in this colony, lingual scraping with disposable plastic sticks was conducted in February during 2 consecutive years (2003 and 2004). The oval, thick-shelled egg containing a larva was found in 15 of 27 arbitrarily-chosen adult monkeys (55.5%) in 2003, and 27 of 106 monkeys (25.5%) in 2004. Infection of other zoo-kept mammals with G. pulchrum was not assessed. Since the gullet worm infects a variety of mammals including primates as natural definitive hosts, and dung beetles and cockroaches as intermediate hosts, the zoological garden may provide an ideal environment for the parasite. Zoo veterinarians should be aware of this disease in kept mammals, and should consider in the case of primates, monthly or bimonthly prophylactic anthelmintic treatment.     

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.     

Identification of an cysteine-to-arginine substitution caused by a single nucleotide polymorphism in the canine monoamine oxidase B gene

Monoamine oxidase B catalytically oxidizes biogenic amines such as phenylethylamine and dopamine, and its activity is presumed to be related to particular behavioral traits. In this study, we first identified a single nucleotide polymorphism (T199C) located on the putative third exon of the canine monoamine oxidase B gene, which causes an amino acid substitution from cysteine to arginine. We then examined the allelic frequencies in five dog breeds (Golden Retriever, Labrador Retriever, Maltese, Miniature Schnauzer, and Shiba) and found significant variation among them. The present results suggest that analysis of the monoamine oxidase B polymorphism could be a useful means of elucidating the genetic background of breed-specific behavioral characteristics in dogs.