Competitive enzyme-linked immunosorbent assay for the detection of the antibodies specific to akabane virus.

A competitive enzyme-linked immunosorbent assay (C-ELISA) using neutralizing monoclonal antibodies (MAbs) against Akabane virus (AKAV) was developed to detect antibodies to AKAV in cattle sera. The performance of the test using 7 different competitor MAbs was evaluated in sequential serum samples and sera from cattle infected with various bovine arboviruses. The dynamics of the antibody response expressed by percentage of inhibition (PI) in C-ELISA coincided with those of neutralizing antibody titers in sequential serum samples from 2 cattle experimentally infected with AKAV. The value of PI in C-ELISA for convalescent sera from cattle infected with arboviruses correlated with the neutralizing antibody titer to AKAV but was unaffected by the antibodies to other arboviruses. In the validation experiment of C-ELISA using 286 bovine sera previously examined for the AKAV antibody by serum neutralization (SN) test, the relative specificity of C-ELISA was more than 98%, whereas the relative sensitivities of individual MAbs ranged from 49% to 82.2%. Overall agreement between C-ELISA and the SN test varied from 72% to 90% depending on the MAb. These results suggest that the C-ELISA is acceptable as a rapid and specific method for detecting antibodies to AKAV and is a potential alternative to the SN test.     

Isolation and chemiluminescent properties of ferret (Mustela putorius furo) polymorphonuclear cells

Ferret polymorphonuclear cells (PMNs) and peripheral blood mononuclear cells (PBMCs) were separated from whole blood by density gradient centrifugation. Using a 50% Percoll solution (density=1.066), PMNs and PBMCs were successfully isolated after centrifugation; the purities of the PMNs and PBMCs were 94.2% and 95.6%, respectively. To evaluate the function of isolated ferret PMNs, we measured the superoxide generation with a MCLA-dependent chemiluminescence assay. The isolated ferret PMNs responded to phorbol 12-myristate 13-acetate (PMA) with kinetics similar to that of human PMNs. The ferret PMNs did not respond to N-formyl-Met-Leu-Phe (fMLF), unlike human PMNs, which rapidly responded. Thus, authors established a method for the rapid separation of highly purified populations of functional PMNs from the whole blood of ferrets.     

Phylogenetic characterization of rabies virus isolates from Carnivora in Brazil

The incidence of canine rabies has been widely reported in Brazil, and new rabies virus (RV) variants, genetically similar to canine RV, have recently been isolated from foxes. In order to derive the epidemiological characteristics of Brazilian Carnivora RV, Brazilian RVs isolated from dogs, cats, and foxes were genetically analyzed. Brazilian Carnivora RV isolates were divided into 2 main lineages. The predominant lineage was found in dogs and cats, which included the Argentinean and Bolivian Carnivora RV isolates, and was extensively distributed throughout Brazil and surrounding countries. The other lineage consisted of three sublineages containing Brazilian dog and fox RV isolates, with the dog sublineages located on an internal branch of 2 fox sublineages, suggesting that RV transmission events might have occurred between foxes and dogs in the past. These results suggest that contact between dogs and wildlife has the potential to generate new rabies variants and that it is important to control RV infection cycles in both dogs and wildlife to prevent spread of rabies infection.     

Malignant pinealoma observed in the deep cerebral parenchyma of a male Wistar rat

This report describes a case of spontaneous malignant pinealoma in a 90-week-old male Wistar rat. The tumor mass occurred in the deep cerebral parenchyma and no intact pineal gland was observed in the area between the posterior-dorsal median line of the cerebrum and the cerebellum. The tumor was characterized by a large nodular proliferation occupying the central area of the brain, extending from the dorsal surface to the base of the brain, corresponding to the thalamus. The tumor cells had round to irregular oblong nuclei approximately 5–17 μm in diameter and showed faintly or moderately eosinophilic cytoplasm and indistinct cell boundaries. Immunohistochemically, the tumor cells were positive for synaptophysin and partially positive for neuron-specific enolase (NSE). The tumor showed malignant features including cellular pleomorphism, high mitotic index, necrotic foci, and invasive and extensive growth and was, therefore, diagnosed as an extremely rare malignant pinealoma in the deep cerebral parenchyma.     

Fruit antioxidant activity, ascorbic acid, total phenol, quercetin, and carotene of Irwin mango fruits stored at low temperature after high electric field pretreatment

Greenhouse-grown tree ripe (TR) and mature green (MG) mangoes (cv. Irwin) were exposed to high electric field treatment before 20 and 30 days of storage at 5 degrees C. MG fruits were allowed to ripen at room temperature after low-temperature storage. Fruit physical quality attributes, ascorbic acid, carotene, quercetin, total phenols, and antioxidant capacity were estimated before and after the storage period. Antioxidant capacity of fruit juice was estimated using the ferric reducing antioxidant power (FRAP) assay. Fruit firmness decreased significantly during storage. Titratable acidity decreased 20 days after storage. Total soluble solids did not change during storage. Antioxidant capacity of fruits remained unchanged up to 20 days of storage period and decreased thereafter. Total phenol and carotenes increased during storage. Antioxidant capacity of fruits was significantly correlated only to ascorbic acids. Peel color and carotenes were higher in TR fruits, whereas titratable acidity and firmness were higher in MG fruits. There was no significant difference in other parameters between the stages of picking. Electric field pretreatment affected the respiration and antioxidant capacity of TR fruits and did not have any significant affect on other parameters. TR mangoes of cv. Irwin are more suitable for low-temperature storage and can be successfully stored for up to 20 days at 5 degrees C without any significant losses in functional properties and quality attributes.     

Maternal Liver Damage Delays Meiotic Resumption in Bovine Oocytes through Impairment of Signalling Cascades Originated from Low p38MAPK Activity in Cumulus Cells

The main objective of the present study is to investigate the molecular mechanism underlying the delay in progression of nuclear maturation in oocytes derived from cows with damaged livers (DL cows), which was previously reported. In present study, delayed progression of nuclear maturation of oocytes derived from DL cows relative to oocytes derived from cows with healthy livers (HL cows) was accompanied by low maturation promoting factor (MPF) activity (0.43 fold, p < 0.05). When cumulus cells were removed from cumulus‐oocyte complexes and the denuded oocytes were cultured, there was no difference in the progression of nuclear maturation between the two liver conditions. In addition, gap junctional communication (GJC) between the oocyte and cumulus cells was higher in DL cows than in HL cows at 3 and 7 h of in vitro maturation (IVM) (p < 0.05). Supplementation of IVM medium with epidermal growth factor (EGF) increased the ratio of germinal vesicle breakdown (GVBD) of oocytes derived from DL cows to the level seen in oocytes derived from HL cows. Additionally, the level of p38MAPK phosphorylation at 0 h of IVM was significantly lower in cumulus cells derived from DL cows than in cumulus cells derived from HL cows (HL cows, 53.5%; DL cows, 28.9%; p < 0.05). Thus, a low level of p38MAPK phosphorylation in cumulus cells induced slow GJC closure between oocyte and cumulus cells, which resulted in slow meiotic maturation of oocytes derived from DL cows.     

Peripartum back fat thickness of multiparous Holstein-Friesian cows with displacement of the abomasum or ketosis

To establish a method to predict postpartum diseases using prepartum back fat thickness (BFT), the peripartum BFTs of 54 healthy multiparous cows before calving, which were diagnosed with postpartum displacement of the abomasum (DA), clinical ketosis or subclinical ketosis were compared with those of healthy cows from 8 weeks before the expected calving date to 8 weeks after calving. The peripartum BFTs of the cows with DA or clinical ketosis were significantly higher than those of healthy cows. The peripartum BFTs of the cows with subclinical ketosis were not significantly higher than those of the healthy cows.     

An index for evaluating the flood-prevention function of paddies

Urbanization is expanding in and around small to medium-sized cities as well as in large cities, and the expansion includes low-lying areas where the flood risk is high. Flood risk in small watersheds has been evaluated using precise mathematical models. However, these models cannot be applied to larger basins due to the complexity of data collection and analysis. As a result, quantitative evaluations of flood risk at macro scales and the flood-prevention function of paddies around urban areas have not been made. This paper proposes a method and an index that can be used to evaluate the flood-prevention function of paddies on a regional scale. In addition, the use of this system for basin-wide management is illustrated by an example for the Kinu and Kokai rivers, which are tributaries of the Tone River of Japan. The applicability of the approach is evaluated using data from a major flood that occurred in the study area.     

Genetic and phylogenetic analysis of glycoprotein of rabies virus isolated from several species in Brazil

Genetic and phylogenetic analyses of the region containing the glycoprotein (G) gene, which is related to pathogenicity and antigenicity, and the G-L intergenic region were carried out in 14 Brazilian rabies virus isolates. The isolates were classified as dog-related rabies virus (DRRV) or vampire bat-related rabies virus (VRRV), by nucleoprotein (N) analysis. The nucleotide and amino acid (AA) homologies of the area containing the G protein gene and G-L intergenic region were generally lower than those of the ectodomain. In both regions, nucleotide and deduced AA homologies were lower among VRRVs than among DRRVs. There were AA differences between DRRV and VRRV at 3 antigenic sites and epitopes (IIa, WB+ and III), suggesting that DRRV and VRRV can be distinguished by differences of antigenicity. In a comparison of phylogenetic trees between the ectodomain and the area containing the G protein gene and G-L intergenic region, the branching patterns of the chiropteran and carnivoran rabies virus groups differed, whereas there were clear similarities in patterns within the DRRV and VRRV groups. Additionally, the VRRV isolates were more closely related to chiropteran strains isolated from Latin America than to Brazilian DRRV. These results indicate that Brazilian rabies virus isolates can be classified as DRRV or VRRV by analysis of the G gene and the G-L intergenic region, as well as by N gene analysis.     

Perceptual learning incepted by decoded fMRI neurofeedback without stimulus presentation

It is controversial whether the adult primate early visual cortex is sufficiently plastic to cause visual perceptual learning (VPL). The controversy occurs partially because most VPL studies have examined correlations between behavioral and neural activity changes rather than cause-and-effect relationships. With an online-feedback method that uses decoded functional magnetic resonance imaging (fMRI) signals, we induced activity patterns only in early visual cortex corresponding to an orientation without stimulus presentation or participants' awareness of what was to be learned. The induced activation caused VPL specific to the orientation. These results suggest that early visual areas are so plastic that mere inductions of activity patterns are sufficient to cause VPL. This technique can induce plasticity in a highly selective manner, potentially leading to powerful training and rehabilitative protocols.