Increased expression of fractalkine and its receptor CX(3)CR1 in canine inflammatory bowel disease and their possible role in recruitment of intraepithelial lymphocytes

The interaction between fractalkine/CX(3)CL1 and its receptor CX(3)CR1 has been reported to play an important role in various human inflammatory diseases, including inflammatory bowel disease (IBD) mediated by lymphocyte chemoattraction. The objective of this study was to investigate the role of fractalkine and CX(3)CR1 in lymphocyte migration in canine IBD. IBD was diagnosed in 34 dogs, and 19 healthy beagles were used as normal controls. We quantified intestinal mRNA and protein expression of fractalkine and CX(3)CR1 by real-time RT-PCR and ELISA, respectively, and examined the localization of fractalkine in canine intestine by immunohistochemistry. The expression of CX(3)CR1 and surface antigens on peripheral blood mononuclear cells (PBMCs) and intraepithelial lymphocytes (IELs) was analyzed by flow cytometry. Intestinal fractalkine and CX(3)CR1 mRNA was significantly up-regulated in IBD dogs compared with the healthy control dogs. In addition, fractalkine expression on intestinal epithelial cells was significantly increased in the intestinal mucosa of IBD dogs compared with the healthy dogs. CX(3)CR1(+) PBMCs were significantly elevated in IBD dogs and positively correlated with the histopathological severity of IELs infiltration. These CX(3)CR1(+) PBMCs predominantly expressed markers for cytotoxic T cells. Almost all IELs expressed CD3, and the majority of cells expressed CD8 rather than CD4, which was analogous to the CX(3)CR1(+) PBMCs. These results suggest that the fractalkine-CX(3)CR1 interaction may contribute to the pathogenesis of canine IBD through migration of IELs.     

Pneumocystis carinii pneumonia in a Cavalier King Charles Spaniel

Pneumocystis carinii pneumonia was diagnosed by postmortem examination of a one-year-old Cavalier King Charles Spaniel with four-week history of dyspnea. Cytologic and histologic examination of lung tissues revealed numerous P. carinii trophozoites and cysts, and P. carinii specific DNA was detected by polymerase chain reaction. The dog showed hypogammagloblinemia and extremely low levels of serum IgG. It was considered that P. carinii pneumonia in this case was associated with an immunodeficient condition which has already been reported in Miniature Dachshunds.     

Molecular cloning of canine thymus and activation-regulated chemokine (TARC) gene and its expression in various tissues

Thymus and activation-regulated chemokine (TARC) is known as a functional ligand for CC chemokine receptor 4 (CCR4), which is selectively expressed on Th2 lymphocytes and induces selective migration of the cells to allergic lesions. In this study, we cloned canine TARC cDNA from canine thymus by RT-PCR with rapid amplification of cDNA ends (RACE) method. The canine TARC clone contained a full-length open reading frame encoding 99 amino acids and included four cysteine residues characteristic to CC chemokine family. The canine TARC cDNA showed 77.5%, 67.4%, and 68.5% amino acid sequence similarity with human, mouse and rat homologues, respectively. Expression of TARC mRNA was detected not only in thymus but also in spleen, lymph node, lung and heart of the various normal dog tissues examined. TARC cDNA clone obtained in this study will be useful for further investigation on allergic diseases in dogs.     

Biological effect of tyrosine kinase inhibitors on three canine mast cell tumor cell lines with various KIT statuses

Tyrosine kinase inhibitors (TKIs) can be important in the treatment of canine mast cell tumor (cMCT). Meanwhile, some TKIs have been identified as substrates for ABCB1. The inhibitory effect of four TKIs (axitinib, imatinib, masitinib, and vatalanib) for proliferation and phosphorylation of c-Kit receptor as well as the expression and function of ABCB1 were investigated in three cMCT cell lines (HRMC, VIMC1, and CMMC1). The IC(50) values of the TKIs in HRMC, the only cell line with wild-type KIT, were clearly higher than those in CMMC1 and VIMC1. In HRMC and CMMC1, both the growth and phosphorylation of c-Kit receptor were suppressed proportionally by the TKIs. VIMC1 required higher concentrations for the inhibition of c-Kit receptor phosphorylation than those in cell growth. The treatment with cyclosporine increased the effects of the TKIs on VIMC1 since ABCB1 was expressed in VIMC1. The results indicated that cMCT cell lines harboring wild-type KIT had lower sensitivity to TKIs. The growth of VIMC1 was seemingly reduced by TKIs through the inhibition of other tyrosine kinases than c-Kit receptor. There was little influence of ABCB1 on TKI effects to the proliferation of VIMC1. These results will be helpful to understand the different sensitivity to TKIs in cMCT patients.     

A case of canine multiple inflammatory colorectal polyps treated by endoscopic polypectomy and argon plasma coagulation

Endoscopic polypectomy and argon plasma coagulation (APC) were performed in a refractory case of inflammatory colorectal polyps in a 7-year-old male Miniature Dachshund. Colonoscopic examination revealed a large sessile polyp and multiple diffuse small polyps, localized to the descending colon and rectum. The case showed a poor therapeutic response to prednisolone and cyclosporine. Under anesthesia, piecemeal resections were performed by polypectomy. APC was carried out to cauterize the polyp remnants. After treatment, reduction of the lesions and the improvement in clinical signs were observed, without recurrence of lesions for at least 10 months. Endoscopic treatment by polypectomy and APC is suggested to be a therapeutic option for refractory cases of inflammatory colorectal polyps in dogs.     

Harpinpsta from Pseudomonas syringae pv. tabaci Is Defective and Deficient in Its Expression and HR-inducing Activity

To elucidate the role of harpins produced by Pseudomonas syringae, the corresponding hrpZ gene was isolated from P. s. pv. tabaci. The sequence information revealed that this gene carries a serious mutation with 326 bp lacking in the central region and potentially encodes only 140 N-terminal amino acids because of a frame shift. The investigation of biological properties using recombinant harpin indicated harpin_psta was incapable of inducing HR in both host and nonhost plants. Based on an immunoblot analysis to detect harpin from P. s. pathovars in hrp-inducing medium, the truncated harpin_psta was neither expressed nor secreted into the culture medium. These results suggest that harpin is not the sole determinant of the host-parasite specificity in P. s. pv. tabaci. Received 10 August 2000/ Accepted in revised form 21 December 2000     

Morphological changes and habitat shifts with growth of endangered floodplain fish: Possible adaptations to fluctuating environments

Adaptations that maximise food intake are dependent on food and/or habitat types. However, there have been few studies on the ability of floodplain primary freshwater fish to rapidly change their morphology, especially the internal one, to maximise utilisation efficiency of food resources in response to environmental fluctuations during their life history. We investigated morphological changes of an Acheilognathinae species (Acheilognathus longipinnis) that inhabits creeks in Japan to determine whether body shape variation correlated with environmental characteristics, including water depth, current velocity, vegetation cover and food availability. When shifting from floodwater-level to low-water-level season, the current velocity of the study area increased as the water level decreased, causing a significant increase in periphyton and a significant decline in zooplankton and phytoplankton. Concurrently, during the short period when A. longipinnis grow from the juvenile to the premature stage, changes in internal morphology, that is intestinal elongation, and changes in external morphology such as an increase of body depth were observed. In contrast, intestinal length significantly decreased in the adult stage. Our findings suggest that morphological changes between floodwater-level and low-water-level seasons associated with juvenile to premature development are an adaptation to changes in food availability due to environmental fluctuations. Additionally, the shortening of the intestine from the premature to the adult stage may be an adaptive strategy for reproduction.     

Differences in bird species diversities between a natural mixed forest and a coniferous plantation

Bird species diversity and bird species richness were surveyed in a natural mixed forest (mature forest) and in a coniferous plantation (30–40 years old) monthly from 1990 to 1994. These forests were 20 km apart and had similar weather conditions and soil types. There were larch and cedar plantations, and hardwoods along streams in the coniferous plantation. In the natural mixed forest, 55 bird species were found, whereas 40 species were recorded in the coniferous plantation. The species diversity of bird communities was significantly higher in the natural mixed forest than in the coniferous plantation mixed with hardwood trees. This result indicates that the coniferous plantation mixed with hardwood trees, presumed to be a more simplified environment, is likely to be less inhabitable for many bird species than the natural mixed forest, suggesting a relationship between the diversity of the forest environment and the diversity of the bird community. A higher number of bird species was recorded in the small coniferous forest mixed with hardwood trees compared with those in the coniferous plantation. The mixing of broad-leaved trees in a coniferous plantation was suggested to be effective in increasing number of bird species. These tendencies were recorded all through the year during our study. In snow-covering periods, both the diversity and the bird species richness fell in all study sites every year. Snow cover would have greater effect on bird species diversity in the coniferous plantation than in other forest types probably due to diet shortages caused by the snow cover over the forest floor.     

Recovery of canopy trees and root collar sprout growth in response to changes in the condition of the parent tree after a fire in a cool-temperate forest

The relationship between the recovery of canopy trees after fire and root collar sprout dynamics was investigated during 1998–2000 in a secondary cool-temperate broad-leaved forest consisting of Quercus mongolica var. grosseserrata and Betula platyphylla var. japonica trees, in northern Hokkaido, Japan, which burned in April 1998. All of the Betula trees that were severely damaged, two-thirds of those slightly damaged, and half of those intact in 1998, died within three growing seasons after the fire. By contrast, half of the Quercus trees that were slightly damaged and half of those severely damaged recovered their foliage, and no slightly damaged or intact trees died during the three growing seasons after the fire. Many Betula trees developed several fruiting bodies of wood-destroying fungi on their stems, irrespective of damage severity. Fungi also infected some of the surviving Quercus, although the crowns tended to recover. Although many sprouting Betula were observed in 1998, the number of sprouts declined rapidly over the study period. Multiple regression analyses showed that the survival and growth of Betula sprouts were positively influenced by the number of sprouts in 1998, damage severity in 1998, and the degree of recover or decline during the study period, and were negatively influenced by parent tree size. On the other hand, a few sprouts of Quercus remained alive. Quercus remained dominant and the dominance of Betula was rapidly reduced after the fire. However, many Betula sprouts remained alive. Stand structure will change drastically for the time being.