Detection of indoleamine 2,3-dioxygenase 1-expressing cells in canine normal and tumor tissues

Cancer immunotherapy is a novel cancer treatment for canine tumors. Indoleamine 2,3-dioxygenase 1 (IDO1) is overexpressed in some human tumors and inhibits antitumor immunity. In this study, we comprehensively evaluated expression pattern of IDO1 and the nature of IDO1-expressing cells in canine normal and tumor tissues. In normal tissue samples, IDO1 expression was detected only in the lymph nodes, spleen, tonsil tissues, and colon tissues. In contrast, IDO1-positive tumor cells were observed in several tumor tissue types. This is the first study to evaluate IDO1 expression in canine normal and tumor tissues, and the results suggest that IDO1 is a promising target for novel cancer immunotherapy in dogs with tumors.     

Pathogenicity, Mating Ability and DNA Restriction Fragment Length Polymorphisms of Pyricularia Populations Isolated from Gramineae, Bambusideae and Zingiberaceae Plants

Eighty-five Pyricularia isolates were collected from 29 host species of Gramineae, Bambusideae and Zingiberaceae plants sampled in Brazil, Uganda, Ivory Coast, India, Nepal, China, Indonesia and Japan. These isolates were compared on the basis of pathogenicity, mating ability and restriction fragment length polymorphisms with single-copy DNA probes. Based on the pathogenicity to eight differential gramineous plants, these isolates were classified into seven pathotypes: finger millet type, foxtail millet type, common millet type, rice type, crabgrass type, Italian ryegrass/ weeping lovegrass type, and non-cereal/grass type. Genetic variation among these isolates was assessed by RFLP analysis with two restriction enzymes and nine single-copy DNA probes isolated from a finger millet strain. An UPGMA dendrogram based on the RFLPs revealed that the 85 isolates could be classified into seven major groups. Isolates from cereal crops (finger millet, foxtail millet, common millet, wheat and rice) and a grass, Brachiaria plantaginea, were clustered into a single group. They were further divided into six subgroups corresponding to the pathotypes. Among cereal crop isolates only an isolate from pearl millet was located into a different group. The remaining isolates were clustered into five groups designated as the crabgrass group, the buffelgrass and jungle rice group, the rice cutgrass, knotroot bristlegrass and Setaria tomentosa group, the bamboo and bamboo grass group and the Zingiber mioga group. The isolates from cereal crops were generally capable of mating with finger millet strains and constituted a closed mating compatibility group. These results suggested that the isolates from cereal crops form a single group with a common ancestor although they are pathogenic to taxonomically diverse plants. A combined analysis of the pathogenicity and genetic similarity suggested that the transmission of M. grisea isolates occurs in natural agroecosystems between finger millet and Eleusine africana, goosegrass or Bambusa arundinacea, between foxtail millet and green bristlegrass, and between rice and tall fescue, Italian ryegrass, sweet vernalgrass, reed canarygrass or Oryza longistaminata. Received 9 August 1999/ Accepted in revised form 12 November 1999     

Serological survey of antibodies to four pathogens in wild boars in Nagano Prefecture,Japan

Wild boars (Sus scrofa) captured or found dead in Nagano Prefecture were surveyed for antibodies to Aujeszky’s disease virus (ADV), Erysipelothrix rhusiopathiae, porcine reproductive and respiratory syndrome virus (PRRSV), and Toxoplasma gondii. While all 168 samples tested were negative for anti-ADV antibodies and all 140 samples tested were negative for anti-PRRSV antibodies, all 190 samples tested were positive for anti-E. rhusiopathiae antibodies and 12 of 180 samples were positive for anti-toxoplasma antibodies. These results suggest that since E. rhusiopathiae and T. gondii cause zoonotic diseases, in addition to wild boars being a potential source of infection for domestic pigs, caution should be taken when handling wild boars or eating wild boar meat because of the possibility of human infection.     

The effect of frequent milk feeding on abomasal curd formation of Holstein calves

In order to understand the effects of the automatic milk replacer feeding system on calf health, we examined the effect of frequent milk feeding on curd formation in the abomasum using ultrasonographic imaging. Eight male Holstein calves were divided into a milk-replacer group and a fresh milk group. Calves were fed twice a day to 12 days after birth (Period A). From 13 days, calves were fed six times a day (Period B). The abomasal fluid was taken by paracentesis. In both periods A and B, the fresh milk group formed bigger curds faster than the milk replacer group. The curd score of the milk replacer group in period B at 2 h was significant lower than those of both groups in period A. The pH in the abomasum was lower in period B than in period A. We could not identify the location of the abomasum in the milk replacer group during period B at almost time points. Our experiment suggests the possibility that frequent feeding of milk replacer causes incomplete hydrolysis of κ-casein as well as curd formation, thus reducing the digestibility compared to the feeding of the milk replacer twice per day or frequent feeding of fresh milk.     

Effect of the d‐glucose analog,d‐allose,on the growth of Arabidopsis roots

Although d ‐glucose increased the root growth of Arabidopsis seedlings, d ‐allose (a d ‐glucose epimer at the third carbon atom) inhibited the root growth at concentrations >0.1 mmol L^?1 and the inhibition increased with increasing d ‐allose concentrations. Allitol (a reduction product of d ‐allose) did not show any significant effect on the growth. The addition of d ‐glucose into the growth medium of Arabidopsis reversed the d ‐allose‐induced growth inhibition, which suggests that the inhibition is not caused by the toxicity of the accumulation of d ‐allose and/or its metabolites in the seedlings. d ‐Allose is phosphorylated by hexokinase, using ATP and phosphate, to allose‐6‐phosphate, with no known capacity for further metabolism. The addition of phosphate into the growth medium did not affect the d ‐allose‐induced growth inhibition and d ‐allose did not reduce the ATP level in the roots. These results suggest that the inhibition is not due to phosphate starvation and ATP depletion. d ‐Mannoheptulose, a specific competitive inhibitor of hexokinase, defeated the d ‐allose‐induced growth inhibition. Hexokinase is known to have a sugar‐sensing function and possibly triggers a signal cascade, resulting in the change of several gene expressions. Therefore, the phosphorylation of d ‐allose by hexokinase might trigger a signal cascade, resulting in the inhibition of Arabidopsis root growth. This is probably a useful model system for studies of the hexokinase‐mediated sugar‐sensing function and for developing new types of weed‐control agents.     

Detection of ruminant meat and bone meal in feeds by sandwich ELISA with monoclonal antibodies

A sensitive and reproducible enzyme-linked immunosorbent assay (ELISA) using two monoclonal antibodies directed against a synthetic peptide with an amino-acid sequence related to the C-terminus of bovine myoglobin and the whole molecule of sodium dodecyl sulphate (SDS)-denatured bovine myoglobin was adapted for detecting bovine myoglobin in contaminated feeds. The ELISA employed bovine meat extract of a known myoglobin concentration as a calibration standard and had an limit of detection (LOD) of 3.54 ng/ml and an limit of quantification (LOQ) of 11.0 ng/ml corresponding to 0.022% and 0.067% (wt/wt) bovine meat-and-bone-meal (MBM) mixed in 20-fold-diluted feed extracts, respectively. A cut-off threshold of 20.6 ng/ml bovine myoglobin was set to simplify ELISA and facilitate quick assessment of test results without a tedious calibration process. The ELISA was able to detect bovine MBM in artificially prepared model feeds, mixed botanical feeds, mixed botanical feeds with skimmed milk, fish meal, pork meal and pork/chicken meal at 0.1% (wt/wt). It was also able to detect sheep MBM in test feeds, but showed no reactivity to swine MBM, chicken MBM, skimmed milk or gelatine of bovine origin. The advantages of this method are the quick and easy extraction protocol of proteins from test feeds, using 100 mM sodium sulphide and 0.6% sodium dodecyl sulphate in the extraction solution and the effective detection of bovine and sheep MBM at 0.1% (wt/wt).     

Expression of podoplanin in various types of feline tumor tissues

Podoplanin is expressed in various human tumors where it promotes tumor progression, epithelial-mesenchymal transition, and distant metastasis. Podoplanin is also expressed in cancer-associated fibroblasts and induces tumor malignancy. The objective of this study was to evaluate podoplanin expression in various types of feline tumor tissues. Immunohistochemical analysis revealed that podoplanin was expressed in cells of 13/15 (87%) squamous cell carcinomas and 5/19 (26%) fibrosarcomas. Moreover, cancer-associated fibroblasts expressed podoplanin in most tumor types, including 18/21 (86%) mammary adenocarcinoma tissues. Our findings demonstrate that various types of feline tumor tissues expressed podoplanin, indicating the importance of the comparative aspects of podoplanin expression, which may be used as a novel research model for podoplanin biology.