Evaluation of a recombinant measles virus expressing hepatitis C virus envelope proteins by infection of human PBL-NOD/Scid/Jak3null mouse

In this study, we infected NOD/Scid/Jak3null mice engrafted human peripheral blood leukocytes (hu-PBL-NOJ) with measles virus Edmonston B strain (MV-Edm) expressing hepatitis C virus (HCV) envelope proteins (rMV-E1E2) to evaluate the immunogenicity as a vaccine candidate. Although human leukocytes could be isolated from the spleen of mock-infected mice during the 2-weeks experiment, the proportion of engrafted human leukocytes in mice infected with MV (10³–10⁵ pfu) or rMV-E1E2 (10⁴ pfu) was decreased. Viral infection of the splenocytes was confirmed by the development of cytopathic effects (CPEs) in co-cultures of splenocytes and B95a cells and verified using RT-PCR. Finally, human antibodies against MV were more frequently observed than E2-specific antibodies in serum from mice infected with a low dose of virus (MV, 10⁰–10¹ pfu, and rMV-E1E2, 10¹–10² pfu). These results showed the possibility of hu-PBL-NOJ mice for the evaluation of the immunogenicity of viral proteins.     

Nitrocapsanthin and nitrofucoxanthin, respective products of capsanthin and fucoxanthin reaction with peroxynitrite

The in vitro reactivity of capsanthin (1) and fucoxanthin (2) with peroxynitrite was investigated, and the reaction products produced by scavenging with peroxynitrite were analyzed. (14'Z)-Nitrocapsanthin (3) and 12-nitrocapsanthin (4) were isolated from the products of the reaction of capsanthin with peroxynitrite. Similarly, (14Z)-15-nitrofucoxanthin (5), (11Z)-11-nitrofucoxanthin (6), and (14Z,9'Z)-15-nitrofucoxanthin (7) were obtained from the reaction of peroxynitrite reaction with fucoxanthin. Capsanthin and fucoxanthin inhibited the nitration of tyrosine by peroxynitrite. Furthermore, nitrocapsanthins (3 and 4) and nitrofucoxanthins (5 and 6) exhibited an inhibitory effect on Epstein-Barr virus early antigen activation in Raji cells and an antiproliferative effect on human pancreatic carcinoma. Moreover, nitrocapsanthins (3 and 4) inhibited carcinogensis of mouse skin tumors initiated by 7,12-dimethylbenz[a]anthracene (DMBN).     

Deoxyribonucleic Acid Homology Among Strains of Soybean-Nodulating Bacteria

Abstract

Characteristics of Brown Forest soils developed under different bio-climatic conditions from low to high eleyations in the Kinki District were studied with special reference to their pedogenetic processes. The Brown Forest soils at high elevations were characterized by a lower bulk density, higher capacities to adsorb organic matter, phosphate, and moisture, which were correlated with the ratio of the amorphous content to the clay content (the value of the ratio of (Feo + Alo)/clay), as compared to those at low elevations. Considering the fact that the value of the (Feo + Alo)/clay ratio was not correlated with the volcanic glass index, the formation of an amorphous fraction at high elevations was considered to proceed according to the following mechanism.

Low temperature at high elevations (above 700 m) may retard the crystallization of oxide minerals. The amorphous oxides with variable positive charge thus formed may adsorb organic matter, confering a darker color and high moisture and high phosphate retention capacities to the subsoil. Adsorbed organic matter stabilizes these amorphous oxides, thus enhancing the amorphous properties and inhibiting crystallization. A1 translocation due to the weak podzolization may contribute to the increase of the content of amorphous materials.

Characterization of the B horizons in forest soils in Japan, in terms of the values of the ratios of (Feo + Alo)/clay, (Fed-Feo)/Fet, and Alo/Ald, (CEC -ECEQ/CEC and STPT-ZPC, suggested that forest soils might be classified into four groups.     

Resinous stem canker development during the growing season of Chamaecyparis obtusa (Hinoki cypress) inoculated with pathogenic fungus Cistella japonica

Seasonal development of resinous stem canker was determined in Chamaecyparis obtusa trees wound‐inoculated with Cistella japonica in January 1996. Samples for anatomical analysis were harvested on 20 May, 9 July and 6 August in the second year of inoculation, and the sections were observed under light microscope. Resin exudation was most abundant in the second year, decreasing in the third. All new resin exudation in the second year was recognized at the May observation. New traumatic resin canal formation was observed in the August samples, and an expansion of necrotic lesions was observed mainly in summer samples in the inoculated trees and in naturally affected trees. Cistella japonica was re‐isolated from all bark lesions or inoculated wounds examined. These results suggest that the activity of Ci. japonica in the tree affects the seasonal development of resinous stem canker during the growing season of the trees.     

Radical scavenging and singlet oxygen quenching activity of marine carotenoid fucoxanthin and its metabolites

Antioxidant activity of carotenoids is suggested to be one of the factors for their disease preventing effects. Marine carotenoids fucoxanthin and its two metabolites, fucoxanthinol and halocynthiaxanthin, have been shown to exhibit several biological effects. The antioxidant activities of these three carotenoids were assessed in vitro with respect to radical scavenging and singlet oxygen quenching abilities. The 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity of fucoxanthin and fucoxanthinol was higher than that of halocynthiaxanthin, with the effective concentration for 50% scavenging (EC 50) being 164.60, 153.78, and 826.39 microM, respectively. 2,2'-Azinobis-3-ethylbenzo thizoline-6-sulphonate radical scavenging activity of fucoxanthinol (EC 50, 2.49 microM) was stronger than that of fucoxanthin (EC 50, 8.94 microM). Hydroxyl radical scavenging activity as measured by the chemiluminescence technique showed that the scavenging activity of fucoxanthin was 7.9 times higher than that by fucoxanthinol, 16.3 times higher than that by halocynthiaxanthin, and 13.5 times higher than that by alpha-tocopherol. A similar trend was observed when the hydroxyl radical scavenging was assessed by the electron spin resonance (ESR) technique. ESR analysis of the superoxide radical scavenging activity also showed the superiority of fucoxanthin over the other two carotenoids tested. Singlet oxygen quenching ability of the three carotenoids was lower than that of beta-carotene, with quenching rate constants ( k Q, x10 (10) M (-1) s (-1)) being 1.19, 1.81, 0.80, and 12.78 for fucoxanthin, fucoxanthinol, halocynthiaxanthin, and beta-carotene, respectively. The higher radical scavenging activity of fucoxanthin and fucoxanthinol compared with halocynthiaxanthin is assumed to be due to presence of the allenic bond.     

Characterization of monoclonal antibodies directed against the canine distemper virus nucleocapsid protein

We have established four monoclonal antibodies (MAbs) against the nucleocapsid protein (NP) of canine distemper virus (CDV). A competitive binding assay has revealed that the MAbs are directed against two antigenic domains. An immunofluorescence assay using a series of deletion clones of the NP and an immunoprecipitation assay using the NP have revealed that two of the MAbs recognize the C-terminal region of the NP while the other two recognize the tertiary structure of the N-terminal domain. These MAbs reacted with all eight strains of CDV used in this study, but showed different reactivities against measles virus and rinderpest virus.     

Cytotoxic T-lymphocyte activity specific for hemagglutinin (H) protein of canine distemper virus in dogs

Cytotoxic T-lymphocyte (CTL) responses to hemagglutinin (H) protein of canine distemper virus (CDV) were evaluated in dogs using the replication-deficient adenovirus protein expression system. Skin fibroblasts were isolated from two dogs and were infected with recombinant adenovirus bearing the CDV-H gene (Ade-CDVH). CTL assay was performed using fibroblasts expressing CDV-H protein as target cells and peripheral blood lymphocytes (PBL) collected from the same dogs one week after immunization of CDV as effector cells. Specific cytotoxic activity was observed against autologous but not heterologous fibroblasts expressing CDV-H protein. These results indicate that the CTL epitope(s) were localized in the H protein.     

Ganglioglioma in the thalamus of a puppy

A solitary brain mass of a 4-month-old miniature dachshund showing seizure-like neurological signs was examined histopathologically. At necropsy a white tumor mass, replacing the thalamus, approximately 1.5 cm in diameter, was found. There was cystic space filled with yellowish pale fluid in the central area of the tumor mass. Histopathological examination revealed that the mass consisted of irregularly arranged well-differentiated neuronal and glial cells, and multifocal mineral deposits. The neuronal cells had a large clear nucleus and various amount of Nissl substances in the cytoplasm. Some neural cells were bi-nucleated. Neither mitotic figures nor proliferating cell nuclear antigen (PCNA)-positive nuclei was found in the neuronal cells. Immunostaining for glial fibrillary acidic protein (GFAP) revealed diffuse proliferation of GFAP-positive glial cells and their processes, while these glial cells did not show apparent cellular atypism, mitotic activity, or PCNA-immunoreactivity. Accordingly, the present tumor was diagnosed as ganglioglioma, and hamartomatous histogenesis might be possible.     

Bovine leukemia virus genotype surveillance in cattle at a slaughterhouse in Aichi Prefecture,Japan, in 2019 using polymerase chain reaction combined with restriction fragment length polymorphism

Polymerase chain reaction (PCR) combined with restriction fragment length polymorphism (RFLP) is commonly used for genotyping bovine leukemia virus (BLV) in slaughterhouses. However, unclassified BLV genotypes have been sporadically reported. To assess the current status of BLV genetic characterization in cattle, PCR-RFLP was performed on blood samples of 170 cattle (84 Japanese Black, 60 Japanese Black x Holstein, and 26 Holstein) from 17 farms (5 prefectures) at a slaughterhouse in Aichi Prefecture in 2019. A total of 65 samples (38.2%) were BLV positive, and genotype 1 was the most predominant (56/65 samples), followed by genotypes 3 (6 samples) and 5 (1 sample), and two unclassified samples. No relationship between the genotypes and breeds was observed. Sequence and phylogenetic analyses demonstrated that unclassified BLV genotypes clustered with genotype 1 sequences were, therefore, not new genotypes.     

Density and sound-speed contrasts,and target strength of Japanese sandeel <Emphasis Type="Italic">Ammodytes personatus</Emphasis>

Sound-speed and density contrasts (h and g, respectively), important acoustic material properties, of Japanese sandeel Ammodytes personatus were measured to estimate theoretical target strength (TS). The measured sound-speed contrast of adult fish varied between 1.016 and 1.023 (mean 1.020), and showed temperature dependence. The measured density contrast differed significantly between juvenile and adult. The density contrast of juvenile varied between 1.017 and 1.024 (1.021), and that of adult varied between 1.026 and 1.038 (1.032). Using these results, TS at 38 and 120 kHz in the fishing season were estimated by an empirical sound scattering model. TS of an individual fish varied significantly with change of tilt angle. TS of near-dorsal aspect (TS_max) and tilt-averaged TS (TS_ave) differed by up to 7 dB. At both frequencies, two different TS_ave−length relationships (TS_ave = a log L + b) were obtained for adult and juvenile. The coefficients of log L of adult were close to 20, suggesting that backscattering strength was proportional to square of body length. These values were larger in juvenile (34.0 at 120 kHz, 56.5 at 38 kHz), suggesting that backscattering strength varied drastically with the cube or fifth power of body length.