Preliminary immunohistochemical study of natriuretic peptide receptor localization in canine and feline heart

We examined the immunohistochemical distributions of natriuretic peptide receptor (NPR)-A, -B and -C that bind with natriuretic peptide hormones A, B and C in four healthy crossbreed young canine and feline cardiac tissues using specific antibodies against human antigens. Cross-immunoreactivities between antigens and antibodies were confirmed using western blot analysis. NPR-A and -C were expressed more strongly in dogs than cats. In both species, these expressions were stronger in the atria than the ventricles, with stronger expression in the left ventricles than the right. NPR-B was largely very weekly or undetected. In canine and feline cardiac tissues, the expressional distribution of NPR-A, -B, and -C closely matched with that of atrial natriuretic peptide, brain natriuretic peptide, and C-type natriuretic peptide as the ligands for corresponding receptors.     

Relation between stockperson behavior toward cows in a holding area and milk yields

To assess the effects of negative and positive behaviors on milk yields, this study examined the attitudes and behaviors of stockpersons toward their cows and milk yields at a farm where 15 workers milked 67 cows as rotating shift work. At a holding area when moving cows to the parlor, stockperson behaviors were observed at 34 milkings. Behaviors were classified as negative (NEG) or positive (POS): NEG were attempts to move cows using touching (NT), vocalization (NV), and gestures (NG); POS were communications without attempting to move cows, consisting of touch (PT) or vocalization (PV). Numbers of NV and NEG were negatively correlated with milk yields of first parity (p < .05). Average milk yields of first parity and all cows were higher when PT were observed (p < .001, p < .0001) and POS were observed (p < .001 and p < .0001). Numbers of PT were positively correlated with milk yields of first parity (p < .05). Average milk yields of second parity were higher when PV was observed (p < .01). Positive behaviors, especially touching, are related to higher herd milk yields. Negative behaviors such as negative vocalizations are related to lower yields of younger cows.     

Growth variability of Pacific saury Cololabis saira larvae under contrasting environments across the Kuroshio axis: survival potential of minority versus majority

Growth variability was examined for Pacific saury Cololabis saira larvae under contrasting environments across the Kuroshio axis, based on samples collected during the winter spawning season in 2013 and 2014. The growth rate index (residual of the otolith marginal 3‐day mean increment width from the linear regression on knob length) of larvae was compared among three areas: the inshore side of the Kuroshio axis, the Kuroshio axis, and the offshore side of the Kuroshio axis in relation to sea surface temperature (SST), salinity (SSS) and chlorophyll‐a (CHL) concentration. The larvae were more densely distributed in the Kuroshio axis and offshore areas of higher temperature and salinity and lower chlorophyll‐a concentration than in the inshore areas of lower temperature and salinity and higher chlorophyll‐a concentration. No marked differences in the growth rate index were found among the three areas, even though the larvae in the inshore areas showed slightly higher growth rates in 2013. Despite the broad ranges of environmental factors, no clear relationship between the growth rate index and any environmental factor was detected. The survival potential of Pacific saury larvae was considered to be at least comparable under contrasting environments across the Kuroshio axis. Such a geographical homogeneity is concluded to be attributable to compensable effects of physical and biological factors. We hypothesize that the minority under physically‐unfavorable but biologically‐favorable conditions on the inshore side of the Kuroshio axis could survive equally well as the majority under physically‐favorable but biologically‐unfavorable conditions around the Kuroshio axis and on the offshore side of the Kuroshio axis.     

Babesia gibsoni rhoptry-associated protein 1 and its potential use as a diagnostic antigen

A cDNA encoding the rhoptry-associated protein 1 (RAP-1) homologue was obtained by immunoscreening an expression library prepared from Babesia gibsoni merozoite mRNA. The complete nucleotide sequence of the gene was 1740bp. Computer analysis suggested that the sequence contains an open reading frame of 1425bp encoding an expected protein with a molecular weight of 52kDa. Based on the sequence similarity, this putative protein was designated as the B. gibsoni RAP-1 (BgRAP-1). The BgRAP-1 gene was expressed in the Escherichia coli BL21 strain, and the recombinant BgRAP-1 was used as the antigen in the enzyme-linked immunosorbent assay (ELISA). The results can differentiate between the B. gibsoni-infected dog sera and the Babesia canis infected dog sera or the normal dog sera. Furthermore, the antibody response against the recombinant protein was maintained during the chronic stage of infection, indicating that the recombinant BgRAP-1 protein might be a useful diagnostic antigen for the detection of antibodies to B. gibsoni infection in dogs.     

A novel culture method of canine peripheral blood lymphocytes with concanavalin a and recombinant human interleukin-2 for adoptive immunotherapy

This study was designed to develop a novel culture method for the efficient proliferation of canine peripheral blood lymphocytes (cPBL) for adoptive immunotherapy. When cPBL were cultured in the presence of concanavalin A (Con A), proliferation of cPBL was induced and expression of interleukin-2 receptor (IL-2R) which enables to respond to exogenously added IL-2 was upregulated. And then, when cPBL were cultured with recombinant human interleukin-2 (rhIL-2) in addition to Con A, proliferation was accelerated and increased to about 10-fold after 1 week. The phenotypic analysis showed that the main population of the cultured cPBL was consisted of CD8+ positive lymphocytes. Among them, CD4+CD8+ double positive (DP) lymphocytes had significantly increased, and the ratio of CD4+ single positive (SP) lymphocytes to CD8+ SP lymphocytes (CD4+SP/CD8+SP) was decreased as compared to before culturing. To evaluate the cytotoxic activity of cPBL cultured with Con A and rhIL-2, furthermore, cytotoxic assay was carried out against xenogeneic melanoma cell line (MeWo), which resulted in MHC-unrestricted cytokilling. These results suggest that the culture method of cPBL by the use of Con A and rhIL-2 may be useful for generating lymphokine activated killer cells, and also this may be beneficial for adoptive immunotherapy of tumor-bearing dogs.     

Vulnerability of feline T-lymphocytes to charged particles

An analysis of ionizing radiation-induced damage in peripheral lymphocytes has been employed to predict the prognosis of radiotherapy in terms of toxicity in normal tissues. Therefore, understanding the sensitivity of lymphocytes to high linear energy transfer (LET)-charged particles would be indispensable for utilizing charged particle therapy in veterinary medicine. However, the availability of such information is very limited. This study aimed to compare the radiosensitivity of feline T lymphocytes to gamma-rays (0.2 keV/microm) and 4 different types of charged particles with LET values ranging from 2.8 to 114 keV/microm. It was observed that the relative biological effectiveness, inactivation cross-section, and isodose-induced apoptosis increased in an LET-dependent manner. On the other hand, no difference in apoptosis frequency was observed in the cells exposed to an isosurvival dose of all the radiation types tested. This is the first study that demonstrates the LET dependence of cell killing and apoptosis induction in feline T lymphocytes. Our results suggest that lymphocytes can be effectively used to predict the prognosis of charged-particle therapy in cat patients.     

Seedborne fungi detected on stored solanaceous berry seeds and their biological activities

We isolated 629 fungi from 1296 berry seeds of solanaceous plants, including tomato (Lycopersicon esculentum), eggplant (Solanum melongena), bell pepper (Capsicum annuum), and red pepper (Capsicum annuum var. annuum) preserved for long and short terms. The isolates were classified into 22 genera excluding unidentified fungi, and the fungal floras were divided into two types: the tomato–eggplant and pepper groups. The results of cluster analysis with unweighted pair-group method with arithmetic average also supported these groups. Most tomato seeds infested with Geotrichum candidum germinated and grew the same as uninfested seeds. Cladosporium sphaerospermum and Arthrinium sp. isolated from eggplant seeds strongly suppressed germination, and Penicillium variabile suppressed seminal root elongation on eggplant. Alternaria alternata, Botrytis cinerea, and Myrothecium verrucaria detected from red pepper or bell pepper seeds were pathogenic to the fruits and the seedlings after artificial inoculation.     

Phytogeny and Cross-infectivity of Powdery Mildew Isolates (Podosphaera fuliginea s. lat.) on Cosmos and Cucumber

To clarify the relationship between the phylogeny and infectivity of isolates of Podosphaera fuliginea s. lat. (= Sphaerotheca fuliginea s. lat.) from cosmos and cucumber, more than 50 powdery mildew isolates from these two plants were subjected to nucleotide sequencing or PCR-RFLP analysis of the rDNA internal transcribed spacer (ITS) region and cross-inoculation tests. The isolates from both cosmos and cucumber are genetically monotypic, and there are six nucleotide substitutions in the rDNA ITS region between isolates from cosmos and cucumber. Cross-inoculation tests of these isolates revealed that isolates from cosmos are not pathogenic on cucumber. Although isolates from cucumber produced conidia on leaves of cosmos in the laboratory, the conidial density was much lower than that from isolates from cosmos. This result, as well as the fact that the cucumber strain was not isolated from cosmos in fields, suggests that isolates from cucumber do not infect cosmos in the field. Therefore, powdery mildews on cosmos and cucumber can be regarded to have become specialized for their hosts both genetically and pathogenically. The present study reconfirms the close relationship between phylogeny and infectivity of powdery mildew fungi. Host specialization may be a trigger that causes genetic divergence of powdery mildew fungi. Received 28 June 2000/ Accepted in revised form 4 September 2000     

Effects of Shengmai San on oxidative damage in mentally stressed mice

AIM: To investigate the preventive effects of Shengmai San (SMS) on oxidative damage in mentally stressed mice.METHODS: An oxidative stress mouse model was established by moustache-removed. Protein carbonyl and thiobarbituric acid reactive substance (TBARS) formation were determined as the oxidative stress markers.RESULTS: (1)Moustache-cut was founded to significantly enhance the behavioral movements of mice, especially large movements (movement 2 and rearing). SMS pre-administration inhibited the accelerated movements. (2) Protein carbonyl was increased in brain, heart, liver and kidney. TBARS in liver and heart increased in the moustache-cut mice, but SMS pretreatment inhibited the increased protein carbonyl and TBARS.CONCLUSION: SMS has the preventive effects on oxidative damage induced by emotional stress.