Induction of dendritic cell-mediated immune responses against canine malignant melanoma cells

To establish the basis for the use of dendritic cells (DC) in the treatment of canine melanoma, dogs were vaccinated using autologous DC pulsed with canine melanoma CMM2 cell lysate in the presence of keyhole limpet haemocyanin (KLH) in vitro (CMM2-KLH-DC), and the induction of immune responses against CMM2 cells in vivo was examined using the delayed-type hypersensitivity (DTH) skin test. The DTH responses against CMM2 cells and KLH were observed in dogs vaccinated with CMM2-KLH-DC, while the responses against KLH but not CMM2 cells were detected with DC pulsed with KLH alone (KLH-DC). Recruitment of CD8 and CD4 T cells was detected in the positively responding sites, suggested that vaccination with CMM2-KLH-DC efficiently elicits T cell-mediated immunity against CMM-2 cells in vivo. These findings demonstrate the potential utility of DC-based tumour vaccination in the treatment of canine malignant melanoma.     

Establishment of a BRAF V595E-mutant canine prostate cancer cell line and the antitumor effects of MEK inhibitors against canine prostate cancer

Canine prostate cancer (cPCa) is a malignant neoplasm with no effective therapy. The BRAF V595E mutation, corresponding to the human BRAF V600E mutation, is found frequently in cPCa. Activating BRAF mutations are recognized as oncogenic drivers, and blockade of MAPK/ERK phosphorylation may be an effective therapeutic target against BRAF-mutated tumours. The aim of this study was to establish a novel cPCa cell line and to clarify the antitumor effects of MEK inhibitors on cPCa in vitro and in vivo. We established the novel CHP-2 cPCa cell line that was derived from the prostatic tissue of a cPCa patient. Sequencing of the canine BRAF gene in two cPCa cell lines revealed the presence of the BRAF V595E mutation. MEK inhibitors (trametinib, cobimetinib and mirdametinib) strongly suppressed cell proliferation in vitro, and trametinib showed the highest efficacy against cPCa cells with minimal cytotoxicity to non-cancer COPK cells. Furthermore, we orally administered 0.3 or 1.0 mg/kg trametinib to CHP-2 xenografted mice and examined its antitumor effects in vivo. Trametinib reduced tumour volume, decreased phosphorylated ERK levels, and lowered Ki-67 expression in xenografts in a dose-dependent manner. Although no clear adverse events were observed with administration, trametinib-treated xenografts showed osteogenesis that was independent of dosage. Our results indicate that trametinib induces cell cycle arrest by inhibiting ERK activation, resulting in cPCa tumour regression in a dose-dependent manner. MEK inhibitors, in addition to BRAF inhibitors, may be a targeted agent option for cPCa with the BRAF V595E mutation.     

Molecular characterization of a novel bovine group A rotavirus

In this study, by partial sequence analysis of the genome segments encoding VP5* and VP7, we characterized a novel bovine group A rotavirus, namely, Tak2, that was detected from adult cattle diarrhea in Tochigi Prefecture, Japan. The nucleotide (nt) and deduced amino acid (aa) sequences of the genome segments encoding VP5* and half of the amino terminal portion of VP7 of Tak2 revealed a low identity with those of group A rotaviruses carrying previously published P and G type specificities (VP5*: nt identity, 61.6%-67.6% and aa identity, 58.0%-71.4%; half of the amino terminal portion of VP7: nt identity, 57.8%-73.5% and aa identity, 61.2%-70.9%). Additionally, phylogenetic analysis of the nt sequences of the genome segments encoding VP5* and half of the amino terminal portion of VP7 revealed that Tak2 formed a branch separate from the established P and G types. These results suggested that Tak2 could possess novel P and G types yet not reported among group A rotaviruses.     

Fishery characteristics of barfin flounder Verasper moseri in southern Tohoku,the major spawning ground,after the start of large-scale stock enhancement in Hokkaido,Japan

Commercial landings of a rare pleuronectid flatfish, barfin flounder Verasper moseri, recovered drastically after large-scale stock enhancement in Hokkaido conducted since 2006. This study investigated commercial landings, fishing grounds, size distributions, and sex and age compositions of barfin flounder during 2007–2011 in southern Tohoku, their major spawning ground, which is over 700 km south of Hokkaido. Landings, mostly comprising stocked fish, increased drastically in southern Tohoku: from 2.0 tons in 2007 to 20.8 tons in 2010. Over 98 % of them were landed during January–April, with the peak period during February–March. Fishing logbook data of offshore bottom-trawl vessels during 2007–2010 revealed the upper continental slope off southernmost Tohoku as the main fishing ground (35°40′–36°50′N, annual weighted mean depth 267–299 m). Two size modes in the landings consisted of males (ca. 40 cm) and females (ca. 60 cm). The main age at fishery recruitment in southern Tohoku was younger for males (age 2+) than for females (ages 3+ and 4+), the result of which was that the female percentage was lowest at 1.5 % in 2009 and highest at 7.9 % in 2011. Our results showed clearly how the effectiveness of large-scale stock enhancement in Hokkaido extended to southern Tohoku in association with spawning migration of stocked barfin flounder.     

Estimating water use of a black locust plantation by the thermal dissipation probe method in the semiarid region of Loess Plateau, China

Black locust (Robinia pseudoacacia) is a major reforestation species in the semiarid region in the Loess Plateau of China. There has been increasing concern about the sustainability of the plantations because of their possible high water-use. This study was, accordingly, undertaken to quantify the stand-scale water use of a middle-aged black locust plantation in the region. The thermal dissipation probe method was applied to 27 trees to measure sap flux densities in an experimental plot during the growing season of 2008. The monoculture stand has a basal area of 23.3 m² ha^?1 and a maximum plant area index (PAI) of 2.89. Sapwood areas were estimated by use of a regressive relationship with the diameter at breast height (DBH) for scaling up of stand transpiration. The results showed that DBH could be a good predictor of sapwood area of individual trees. The diurnal cycles of average sap flux densities differed among DBH classes. Daily transpiration can be predicted from mean daily daytime vapor pressure deficit (VPD_m) using a fitted exponential saturation model. Model variables were different among seasons, probably owing to different soil water conditions and leaf phenology. By using the derived model for each month, stand canopy transpiration over the growing season was estimated to be 73.8 mm, with an average daily value of 0.41 mm day^?1 and a maximum of 0.89 mm day^?1. The relatively small estimates of stand transpiration might be attributed to low PAI and sap wood area of the middle-aged stand.     

Relationship between annual rainfall and interception ratio for forests across Japan

Several previous studies in Japan have examined differences in rainfall interception amounts induced by differences in forest properties by comparing the annual rainfall interception ratios (annual rainfall interception divided by annual rainfall) from various sites without considering variations in meteorological conditions between sites. Rainfall interception actually depends on meteorological conditions as well as forest properties. This study examined variations in the annual interception ratio relating to the variation in annual rainfall, which would be the primary factor relating to the interception ratio, across Japan with the use of a rainfall interception model assuming the same forest properties (i.e., the canopy storage capacity, canopy closure, leaf area index (LAI), and the bulk coefficient for sensible heat transfer). The ratio ranged between 0.12 and 0.24 across Japan and was highly correlated to annual rainfall. This indicates that considering the variation in annual rainfall is critical for assessing the difference in rainfall interception amounts induced by forest properties. We reconsidered the results of previous studies in Japan that compared annual interception ratios between sites with different forest properties: (i) there is no clear difference in interception amounts between broadleaf and coniferous forests and (ii) there is a positive correlation in stem density and interception amounts for coniferous forests. These results still held when considering differences in annual rainfall between sites.     

Evaluation of leaf display of evergreen broadleaved tree species and deciduous tree species in warm temperate conifer plantations

We investigated the sapling leaf display in the shade among trees of various leaf lifespans co-occurring under the canopy of a warm-temperate conifer plantation. We measured leaf-area ratio (aLAR) and morphological traits of saplings of evergreen broadleaved tree species and a deciduous tree species. Although we found large interspecific and intraspecific differences in aLAR even among saplings of similar size in the homogeneous light environment, we did not find a consistent trend in aLAR with leaf lifespan among the species. While deciduous trees annually produced a large leaf area, some evergreen broadleaved trees retained their leaves across years and had aLAR values as high as those of deciduous trees. Among leaf-level, shoot-level, and individual-level morphological traits, aLAR was positively correlated with current-year shoots mass per aboveground biomass in deciduous trees, and with the area of old leaves per aboveground mass in evergreen broadleaved trees. Thus, tree-to-tree variation in the degrees of annual shoot production and the accumulation of old leaves were responsible for the interspecific and intraspecific variations in aLAR.     

Effect of forest structure on the spatial variation in soil respiration in a Bornean tropical rainforest

This study was undertaken to identify critical and practical factors explaining spatial variations in soil respiration and to estimate stand-scale soil respiration in an aseasonal tropical rainforest on Borneo Island. To this aim, we conducted soil respiration measurements at 25 points in a 40 m × 40 m subplot of a 4 ha study plot between 2002 and 2006, and examined the spatial variation in soil respiration averaged over the 4 years in relation to soil, root, and forest structural factors. In addition, we examined the spatial representativeness of soil respiration measured in the subplot using a specific scaling procedure. Consequently, we found significant positive correlation between the soil respiration and forest structural parameters such as the mean diameter at breast height (DBH), total basal area, and maximum DBH within 6 m of the measurement points. The most important factor was the mean DBH within 6 m of the measurement points, which had a significant linear relationship with soil respiration. Using the derived linear regression and an inventory dataset, we estimated the 4 ha plot-scale soil respiration. The 4 ha plot-scale estimation (6.0 μmol m⁻² s⁻¹) was nearly identical to the subplot-scale measurements (5.7 μmol m⁻² s⁻¹), which were roughly comparable to the nocturnal CO₂ fluxes calculated using the eddy covariance technique. In addition, we discuss characteristics of the stand-scale soil respiration at this site by comparing with those of other forests reported in previous literature in terms of the soil C balance. Soil respiration at our site was noticeably greater, relative to the incident litterfall amount, than soil respiration in other tropical and temperate forests probably owing to the larger total belowground C allocation by emergent trees. Overall, this study suggests the arrangement of emergent trees with larger DBH and their belowground C allocation could be primary factors controlling spatial variations in soil respiration in the tropical rainforest.     

An interpretation of the conductometric titration curve of humic acid

Methods of fractionating the functional groups of humic acid by conductometric titration were examined. Replacement titration, in which a sample solution is titrated with a standard acid solution after adding an excess amount of a standard base, seems to be a suitable method. By this method, the functional groups of humic acid were fractionated into three clusters of different dissociation constants, designated as VW (very weak), W (weak), and S (strong) clusters. The VW cluster was presumed to include phenolic hydroxyl residues, carboxyl residues and nitrogenous bases of very high pK's. The W cluster was presumed to include various aliphatic and aromatic carboxyl residues and nitrogenous bases of medium pK's, whereas the S cluster was presumed to include carboxyl residues of low pK₁ of salicylate or phthalate. A humic acid sample obtained from the A horizon of Shitara “kuroboku” soil had 0.78 me/g of the VW cluster, 3.24 me/g of the W cluster and 1.90 me/g of the S cluster.     

Horizontal movement and emigration of juvenile spotted halibut <Emphasis Type="Italic">Verasper variegatus</Emphasis> released in a shallow brackish lagoon: Matsukawa-ura,northeastern Japan,revealed by acoustic telemetry

To elucidate the post-release movement and emigration of juvenile spotted halibut Verasper variegatus, ten hatchery-reared age-0 individuals (10 months old, 17.0–19.7 cm total length) surgically implanted with acoustic transmitters were released near the center of Matsukawa-ura, a shallow brackish lagoon, on 25 November 2009 (water temperature 13.8 °C). They were monitored by ten acoustic receivers for 5 months. Of ten individuals, eight left the release site during December–March. Three of these emigrated to the outer ocean on 17 and 29 January and 30 March 2010, respectively. Juveniles probably started wintering migration in December and January when water temperatures decreased considerably in the lagoon (mean 9.5 and 6.0 °C, respectively). They stopped their migration in the coldest month, February (mean 5.7 °C), and restarted it in March when water temperatures frequently exceeded a plausible threshold for movement (≥6 °C). Statistical analyses revealed that the fish started migration significantly more frequently at nighttime. The migration track of an individual recorded from 11 to 30 March showed gradual nocturnal movements and a slow migration speed (estimated maximum speed 2.2 m/min). Our results revealed that water temperature primarily governed the seasonal timing of nocturnal migratory movements of juvenile spotted halibut.