Expression of alpha-hemoglobin stabilizing protein and cellular prion protein in a subclone of murine erythroleukemia cell line MEL

Alpha-Hemoglobin stabilizing protein (AHSP) functions as the erythroid-specific molecular chaperon for alpha-globin. AHSP gene expression has been reported to be downregulated in hematopoietic tissues of animals suffering from prion diseases though the mechanism remains to be clarified. Herein, we demonstrate that MELhipod8 cells, a subclone of murine erythroleukemia (MEL) cells, have prion protein (PrPc) on the cell surface and have highly inducible expression of the AHSP and alpha- and beta-globin genes, resembling the expression pattern of the PrP and AHSP genes in bipotential erythroid- and megakaryocyte-lineage cells followed by erythroid differentiation in normal erythropoiesis. Moreover, MELhipod8 cells exhibit greater effective erythroid differentiation with a population of hemoglobinized normoblast-like cells than that observed for the parental MEL cells. These findings suggest that MELhipod8 cells could provide a mechanism for downregulation of the AHSP gene in prion diseases.     

FCV-VBS isolated from cats with typical symptoms caused VSD in experimental cats

Commercially available vaccines have been used widely to prevent feline calicivirus infection (FCI). However, with their widespread use, field strains, which are weakly cross-reactive with the live-virus vaccine strain F9, have posed the problem of vaccine breakdown. Recently the existence of FCV—associated virulent systemic disease (VSD) has been published. But their molecular diversity, antigenic mutations and physicochemical property have not been sufficiently clarified. Thus, we experimentally gave the vaccine breakdown strain (VBS) H10 to cats that had been inoculated with an F9 live vaccine. After the administration of strain H10, vaccinated cats (1 through 4) had no respiratory symptoms, whereas the non-vaccinated cat 5 showed clinical symptoms such as a fever of over 40°C, loss of vitality, decreased appetite, diarrhea, and nasal discharge after receiving strain H10, and died. Lethal FCV is rare, and may be a virulent systemic disease (VSD)—inducing strain. This is the initial report on VSD in Japan. It has been reported that symptoms of VSD were similar in vaccinated and nonvaccinated cats on experimental infection. However, no VSD-like symptoms developed, and the incidence of the disease varied depending on the presence or absence of vaccination, suggesting that there are two mechanisms of vaccine breakdown: one is associated with the vaccine immunity level, and the other is not. The characteristics of the VBS revealed were: (1) the duration of virus excretion was short when the originally carried antibody titer before virus challenge was high, (2) the excreted viral molecular species varied daily, not being limited to a specific species with time, and (3) the acquired physicochemical properties did not persist, and altered daily. FCV-VBS alters the molecular species and physicochemical properties daily due to the reduction of host immunity, which may lead to VSD.     

Pharmacokinetics and toxicity of zonisamide in cats

With the eventual goal of making zonisamide (ZNS), a relatively new antiepileptic drug, available for the treatment of epilepsy in cats, the pharmacokinetics after a single oral administration at 10mg/kg and the toxicity after 9-week daily administration of 20mg/kg/day of ZNS were studied in healthy cats. Pharmacokinetic parameters obtained with a single administration of ZNS at 10mg/day were as follows: Cmax=13.1microg/ml; Tmax=4.0h; T(1/2)=33.0h; areas under the curves (AUCs)=720.3microg/mlh (values represent the medians). The study with daily administrations revealed that the toxicity of ZNS was comparatively low in cats, suggesting that it may be an available drug for cats. However, half of the cats that were administered 20mg/kg/day daily showed adverse reactions such as anorexia, diarrhoea, vomiting, somnolence and locomotor ataxia.     

The effect of approach direction and pace on flight distance of beef breeding cows

The aim of this study is to determine the effect of approach direction and pace on flight distance of beef breeding cows. Five Japanese Black breeding cows were used as experimental cattle. Three approach directions were tested; that were the approach toward the head of the test cow from the front direction (FRONT), the approach toward to the foreleg of the test cow from the right or left (SIDE) and the approach toward to hind‐leg of the test cow diagonally from the right or left rear (BACK). Three levels of approach pace were also tested; that were the approach at a pace of 7 km/h, the approach at a pace of 4 km/h, and the approach at a pace of 1 km/h. The shortest distance between observer and cow's at the moment of the cow's withdrawal was defined as the flight distance. The flight distance in FRONT was significantly shorter than that in SIDE. The effect of the approach pace was significant. The faster an observer approached to cow, the greater the flight distance shown by the cow. The manner of approach to cattle markedly affects the flight distance. Therefore, the manner of approach to cattle must be standardized to measure flight distance for sharing results obtained by several researchers under various conditions.     

Lumenal localization in the endoplasmic reticulum of the C-terminal tail of an AE1 mutant responsible for hereditary spherocytosis in cattle

An R664X nonsense mutant AE1 is responsible for dominant hereditary spherocytosis in cattle and is degraded by the proteasomal endoplasmic reticulum-associated degradation. The present study demonstrated that R664X AE1 translated in vitro had the trypsin-sensitve site identical to that of the wild-type AE1. The P661S/R664X mutant containing a possible N-glycosylation site at Asn660 showed an increase in size by 3 kDa both in the cell-free translation system and in transfected HEK293 cells. Moreover, steady state levels of R664X and P661S/R664X in HEK293 cells were markedly increased in the presence of a proteasome inhibitior. These findings indicate that the truncated C-terminal region of R664X AE1 has lumenal localization in the endoplasmic reticulum and is not accessible to proteasomal machineries in the cytosol.     

Codend selectivity for jack mackerel and whitefin jack and unequal split parameter estimates observed in trouser trawl experiments

Codend selectivity for the jack mackerel Trachurus japonicus and the whitefin jack Kaiwarinus equula were evaluated based on data from trouser trawl experiments carried out in the East China Sea, using a test codend of 60 mm diamond mesh and a control codend made of minnow net with a square mesh of 9 mm bar length. Between-haul variations in parameters and the mean selection curves were tested with the catch data in the SELECT approach, and then the model of between-haul variation in the split parameter with the mean selection curve was chosen as the best fit using Akaike’s information criterion model selection. The 50% retention lengths and the selection ranges were 11.4 and 3.36 cm for jack mackerel and 8.83 and 0.93 cm for whitefin jack, respectively. The selection curve for whitefin jack was sharp, whereas that for jack mackerel was relatively wide. As the estimated split parameters indicated, about 80% of the whitefin jack entered the control codend, but 85 and 90% of the jack mackerel entered the control codend in the second and third hauls, respectively. The inequality in the split parameter is discussed from the viewpoint of the animal’s swimming behavior and water movement based on underwater video observations.     

Metagenomic analysis using 16S ribosomal RNA genes of a bacterial community in an urban stream,the Tama River,Tokyo

In an effort to determine genus- or species-level taxonomic profiles and diversity of bacterial consortia in the Tama River around urban Tokyo, next-generation sequencing technology targeting a 16S ribosomal RNA (rRNA) gene amplicon was employed. Metagenomic analysis performed by an Ion Personal Genome Machine after sequentially filtering samples through 5-, 0.8- and 0.2-μm filters yielded 1.48 Gb of 16S sequences (average 2.38 M reads/sample). The results indicated that half of the bacterial sequences belonged to Proteobacteria, followed by Bacteroidetes, Actinobacteria and Cyanobacteria. Flavobacterium (Bacteroidetes), possibly including a potential fish pathogen, was the most numerous genera in the Tama River metagenome, and accounted for?~?16% of assigned 16S reads, followed by Mycobacterium. Other dominant bacterial genera including Zoogloea, Sediminibacterium, Hyphomicrobium, Sphingopyxis, Thiothrix and Lysobacter, were thought to be associated with waste water and sludge. MiSeq metagenomic analysis revealed that environmental factors, particularly water temperature, influenced the bacterial composition throughout the year, with a strong negative correlation observed for Proteobacteria and a positive correlation for Bacteroidetes. In terms of bacterial genera, Flavobacterium was positively correlated with temperature, while Polaromonas, Pseudomonas and Bradyrhizobium were negatively correlated with this, suggesting dynamic change in the free-living bacterial population throughout the year and versatile adaptation strategies in relation to environmental factors.     

Quantifying growth in maricultured corals using photogrammetry

In coral mariculture, growth is one of the most common ways to track success. However, numerous methods of monitoring coral growth make comparative studies challenging. A literature review of 39 studies from 1982 to 2017 indicated that the most predominant non‐invasive methods used were linear and areal measurement and these were evaluated for their accuracy using nursery‐reared corals. The monthly change in linear and areal growth rates of six coral species (n = 215), Pocillopora acuta, Hydnophora rigida, Merulina ampliata, Podabacia crustacea, Echinopora lamellosa and Platygyra sinensis were measured via photogrammetry. We tested whether the planar area of coral colonies can be estimated using three geometric formulas of linear measurements. Based on the literature review, the six types of measurement methods were namely linear, volume, weight, area, count of polyps/branches and calcification, in decreasing order of application. Our results showed that the change in area calculated by geometric mean diameter (GMD) formulas provided the most accurate estimation among the three formulas and was strongly correlated with planar area (R² ≥ .60; p < .05) for all coral species, except E. lamellosa and Pl. sinensis. However, our findings suggest that it is not ideal to use geometric formulas to estimate the change in area. Instead, we posit that areal photogrammetry represents the simplest yet accurate non‐invasive method for rapid monitoring of extensive areas of corals in situ. Lastly, we discuss the recommendations and limitations for areal photogrammetry.     

Current classification of Ralstonia solanacearum and genetic diversity of the strains in Japan

Ralstonia solanacearum is the causal organism of bacterial wilt of more than 200 species representing 50 families of plants in tropical, subtropical, and warm temperate regions in the world. Traditionally classified into five races based on differences in host range, R. solanacearum has also been grouped into six biovars on the basis of biochemical properties. With recent developments in molecular biology, various DNA-based analyses have been introduced and used to confirm that this binary system does not completely represent the diversity within R. solanacearum strains. Therefore, a new hierarchical classification scheme has been suggested, which defines R. solanacearum as a species complex and reorganized the concept of the species as a monophyletic cluster according to a phylogenetic analysis based on genomic sequence data. Here we discuss the current bacterial wilt situation and genetic relationships based on the recent classification system of Japanese R. solanacearum strains as well as worldwide strains. We also review the genetic, biochemical, and pathological characteristics of R. solanacearum strains, in particular, those affecting potato and Zingiberaceae plants as distinctly important pathogens in relation to continuously problematic and recent emergent diseases in Japan.     

Allometric equations for accurate estimation of above-ground biomass in logged-over tropical rainforests in Sarawak, Malaysia

Although allometric equations can be used to accurately estimate biomass and/or carbon stock in forest ecosystems, few have been developed for logged-over tropical rainforests in Southeast Asia. We developed allometric relationships between tree size variables (stem diameter at breast height (dbh) and tree height) and leaf, branch, stem and total above-ground biomass in two logged-over tropical rainforests with different soil conditions in Sarawak, Malaysia. The study sites were originally classified as mainly lowland dipterocarp forest and have been selectively logged in the past 20 years. In total, 30 individuals from 27 species were harvested to measure above-ground parts. The correlation coefficients for the allometric relationships obtained for total above-ground biomass as a function of dbh had high values (0.99), although the relationships for leaf biomass had a relatively low coefficient (0.83). We also found relatively high coefficients for allometric relationships between tree height and plant-part biomass, ranging from 0.82 to 0.97. Moreover, there were no differences for allometric equations of total above-ground biomass between study sites. A comparison of equations of above-ground biomass in various previously reported tropical rainforests and pan-tropic general equations imply that our allometric equations differ largely from the equations for tropical primary forests, early successional secondary forest, and even for the general models. Therefore, choosing the biomass estimation models for above-ground biomass in the logged-over forests of Southeast Asia requires careful consideration of their suitability.