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.     

First case report of Sarcocystis neurona-induced equine protozoal myeloencephalitis in Japan

Equine protozoal myeloencephalitis developed in a three-year-old male Thoroughbred racehorse imported from the United States. The animal showed astasia five days after the onset of ataxia. Histopathologically, focal nonpurulent myelitis accompanied by hemorrhage and perivascular infiltration was observed in the fourth and fifth cervical spinal cord. Immunohistochemically, shizonts were occasionally observed and were positive for anti-Sarcocystis neurona (S. neurona) antiserum. S. neurona-specific antibodies were detected in the serum and cerebrospinal fluid by Western blot. This is the first equine protozoal myeloencephalitis case in Japan.     

Occurrence of tetrodotoxin in three Nassarius gastropod species in Khanh Hoa Province,Vietnam

Fisheries Science - Neurotoxic poisonings with fatal symptoms caused by consumption of Nassarius gastropods have been reported in Vietnam but the causative toxins have not been confirmed. In the...     

Effects of lectin in the scleractinian coral Ctenactis echinata on symbiotic zooxanthellae

We report herein the presence of a lectin in the scleractinian coral Ctenactis (Fungia) echinata. The lectin bound preferentially to lactose, melibiose, and d-galactose. The purified lectin CecL was composed of several isolectins, and it was found to have a molecular mass of 67.4 kDa via gel filtration. Glycopeptidase F-treated CecL showed a single band at 32.5 kDa. The mass/charge ratios of the reduced CecL peaks were equivalent to half those of the native peaks. These results suggest that CecL is composed of two glycosylated polypeptides linked by interchain disulfide bonds. In a biological activity test using a zooxanthellal culture (Dinoflagellate Symbiodinium) clonally isolated from Fungia cf. fungites, CecL transformed the flagellated motile form of Symbiodinium into the nonmotile coccoid form, a form equivalent to the symbiotic stage. The activity of CecL on Symbiodinium cells was concentration dependent, and 100 μg/ml CecL arrested Symbiodinium cells in the coccoid form for 5 days. CecL also suppressed the growth of Symbiodinium cells, unlike the octocoral lectin derived from Sinularia lochmodes, which arrests Symbiodinium cells in the coccoid form but does not affect the growth of the coccoid. This result provides further evidence that coral lectins play a role in symbiont engagement and maintenance in zooxanthellae–coral symbiosis.     

Characterization of gelatinolytic enzymes in the skeletal muscle of red sea bream Pagrus major

Gelatinolytic enzymes were partially purified from the skeletal muscle of red sea bream Pagrus major and characterized to obtain information on post mortem tenderization of fish muscle. Four gelatinolytic activities, G1 (90 kDa), G2 (65 kDa), G3 (60 kDa), and G4 (100 kDa), were detected in the Q Sepharose column. G1, the major gelatinolytic enzyme, and G4 were identified as serine proteinases from results of inhibitor spectrum and substrate specificity. By contrast, G2 and G3 were found to be metalloproteinases since these were inhibited by ethylenediamine tetraacetic acid and o-phenanthroline, and activated by 4-aminophenylmercuric acetate. The optimum pH and temperature of these enzymes were in the ranges of 7–9 and 20–40°C, respectively.     

Evaluation and improvement of MODIS gross primary productivity in typical forest ecosystems of East Asia based on eddy covariance measurements

Gross primary productivity (GPP) is a major component of carbon exchange between the atmosphere and terrestrial ecosystems and a key component of the terrestrial carbon cycle. Because of the large spatial heterogeneity and temporal dynamics of ecosystems, it is a challenge to estimate GPP accurately at global or regional scales. The 8-day MODerate resolution Imaging Spectroradiometer (MODIS) GPP product provides a near real time estimate of global GPP. However, previous studies indicated that MODIS GPP has large uncertainties, partly caused by biases in parameterization and forcing data. In this study, MODIS GPP was validated using GPP derived from the eddy covariance flux measurements at five typical forest sites in East Asia. The validation indicated that MODIS GPP was seriously underestimated in these forest ecosystems of East Asia, especially at northern sites. With observed meteorological data, fraction of photosynthetically active radiation absorbed by the plant canopy (fPAR) calculated using smoothed MODIS leaf area index, and optimized maximum light use efficiency (ε _max) to force the MOD17 algorithm, the agreement between predicted GPP and tower-based GPP was significantly improved. The errors of MODIS GPP in these forest ecosystems of East Asia were mainly caused by uncertainties in ε _max, followed by those in fPAR and meteorological data. The separation of canopy into sunlit and shaded leaves, for which GPP is individually calculated, can improve GPP simulation significantly.     

Mechanical stretch-induced activation of skeletal muscle satellite cells is dependent on nitric oxide production in vitro

Mechanical stretch induces activation of cultured quiescent satellite cells and the activation response is owing to rapid release of hepatocyte growth factor (HGF) from its extracellular association with satellite cells and its subsequent presentation to the c-met receptor. We provide new evidence that the stretch activation is dependent on nitric oxide (NO) production. Stretch activation could be abolished by the addition of N ^G-nitro- L -arginine methyl ester (L-NAME), a competitive inhibitor of NO synthesis, but not by N ^G-nitro- D -arginine methyl ester hydrochloride, a less active enantiomer of L-NAME. Adding HGF to the L-NAME culture restored the activation response, indicating that L-NAME does not directly inhibit satellite cell activation, but acts upstream from the HGF release. In addition, immunoblots of satellite cell lysate revealed the presence of nitric oxide synthase. These experiments suggest that NO is involved in linking mechanical perturbation of satellite cells to chemical signaling responsible for HGF release from its sequestration in vitro .     

Preparation of calibration standards of N1-H paralytic shellfish toxin analogues by large-scale culture of cyanobacterium Anabaena circinalis (TA04)

Mouse bioassay is the official testing method to quantify paralytic shellfish toxins (PSTs) in bivalves. A number of alternative analytical methods have been reported. Some methods have been evaluated by a single laboratory validation. Among the different types of methods, chemical analyses are capable of identifying and quantifying the toxins, however a shortage of the necessary calibration standards hampers implementation of the chemical analyses in routine monitoring of PSTs in bivalves. In our present study, we studied preparation of major PST analogues as calibrants by large-scale cultivation of toxic freshwater cyanobacteria Anabaena circinalis TA04. The cells were steadily grown in 10 L bottle for 28 days. The primary N1-H toxins, C1/C2, were produced at a concentration of 1.3 ± 0.1 μmol/L. The intracellular and extracellular toxins occupied 80% and 20%, respectively. Over 220 μmol of the toxins was obtained from approximately 200 L of the culture over six months, demonstrating that it is sufficient to prepare saxitoxin analogues. The toxins were chemically converted to six N1-H analogues. Preparation of the analogues was carried out at relatively high yields (50-90%). The results indicate that our preparation method is useful to produce N1-H toxins. In our present study, detailed conditions for preparation of one of the rare N1-H analogues, gonyautoxin-5, were investigated.