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application/pdf, Veterinary and Comparative Oncology. 2022, 20 (1), P.134-141

A 10-month-old, intact male Toy Poodle was referred for a postural abnormality. Blood biochemical tests revealed a marked increase in plasma creatine phosphokinase (CPK) concentration. The isoenzyme test showed that 99% of serum CPK consisted of CPK-MM. Histopathological evaluation of muscle biopsy samples confirmed scattered degeneration and necrosis of myofibers. Immunohistochemistry for dystrophin showed an absence of staining in muscle cells. Based on these findings, the dog was diagnosed with dystrophin-deficient muscular dystrophy. Whole genome sequencing using genomic DNA extracted from blood revealed a single base pair insertion in exon 45 of the Duchenne muscular dystrophy (DMD) gene. This is the first report on muscular dystrophy in Toy Poodles and identified a novel mutation in the DMD gene.

Human patients with inflammatory bowel disease may have poor prognosis with hypozincemia. However, there are limited data on zinc concentrations in the blood of dogs with lymphocytic-plasmacytic enteritis (LPE). The purpose of this study was to investigate the serum zinc concentration in dogs with LPE and its influence on disease severity and prognosis. Thirty-five dogs with LPE were recruited. Serum zinc concentration was measured using atomic absorption spectrometry. Hypozincemia was observed in 18/35 (51%) dogs with LPE. Serum zinc concentration was inversely correlated with histological and clinical severities. Overall survivals were significantly shorter in dogs with hypozincemia than in those without it. These findings suggest that serum zinc concentration is a useful biomarker for LPE severity and prognosis in dogs.

Leiomyoma is the most common mesenchymal tumor in the gastrointestinal (GI) tract. Leiomyomas usually have a single or multinodular mass of various sizes, and affected animals can develop alimentary symptoms depending on the location and size. A 3-year old female miniature dachshund died after a history of refractory rectal prolapse, esophagectasis and aspiration pneumonia. At necropsy, the GI wall at the gastroesophageal and anorectal junctions was circumferentially thickened. Histologically, both GI lesions were composed of bundles of well-differentiated smooth muscles without mass formation or invasive growth. The neoplastic cells had little cellular atypia and low proliferative activity, and were positive for α-smooth muscle actin. The lesions were diagnosed as diffuse leiomyomatosis with circumferential thickening of the GI wall and has not been described in the veterinary literature.

application/pdf, Journal of Comparative Pathology. 2019, 168, P.8-12

Canine tonsillar squamous cell carcinoma (TSCC) shows a higher metastasis rate than non-tonsillar oral SCC (NTSCC). The mechanisms of metastasis for TSCC have been less studied, because both TSCC and NTSCC cell lines are few. In this study, 6 cloned TSCC (TSCCLN#1-#6), which were from a metastatic lymph node, and 2 cloned NTSCC (oSCC-1 and -4) cell lines, which were from the primary lesion, were established, and their characteristics were evaluated in vitro and in vivo. Results showed that increased expression level of Vimentin in TSCC cell lines and increased expression levels of mesenchymal markers including Vimentin, Snail, and Slug in NTSCC cell lines corelated with the malignant phenotypes such as the cell growth and colony formation abilities in vitro. However, expression levels of mesenchymal markers and in vitro characteristics were unrelated to tumorigenic ability in nude mice. Additionally, the expression levels of E-cadherin and Vimentin were also evaluated by immunohistochemistry using the formalin-fixed paraffin embedded canine oral SCC tissues, and the results show that the expression level of Vimentin in TSCC was higher than in NTSCC. In conclusion, the cell lines established in this study might contribute to elucidating the mechanisms involved in TSCC metastasis.

Ebstein anomaly is a rare congenital heart disease that has been described in domestic dogs, a meerkat, a pygmy goat, and a lion. An 11-mo-old Tsushima leopard cat presented to Osaka Prefecture University Veterinary Hospital for diagnosis and treatment of right-sided congestive heart failure. Echocardiography showed a dilated right atrium and ventricle with an enlarged tricuspid valve annulus and apical displacement of the tricuspid valve leaflets. The cat was diagnosed with Ebstein anomaly. To the best of our knowledge, this is the first report of this type of congenital heart disease in a Tsushima leopard cat.

An 11-year-old female golden retriever dog had a mass at the right corner of the upper lip, which gradually increased in size and protruded into the oral cavity. The mass was removed surgically. The cut surface of the mass was smooth, whitish and solid, and covered by the oral mucosal membrane. Histopathologically, the mass consisted mainly of reticular pattern of short spindle cells that stained positively for cytokeratin AE1/AE3, α-smooth muscle actin and p63, suggestive of a myoepithelial cell phenotype. Between the neoplastic cords, there was myxoid or edematous connective tissue. Additionally, neoplastic cells with luminal epithelial and basal cell phenotypes were arranged in ducts and small islands, respectively. Based on the diverse histological and immunohistochemical features, the tumor was diagnosed as pleomorphic adenoma of the labial gland. To our knowledge, the reticular proliferation pattern of myoepithelial cells has not been described in salivary gland tumors of domestic animals.

We evaluated changes in peripheral blood lymphocyte (PBL) count in dogs following adoptive immunotherapy using lymphokine-activated T killer cells (T-LAK) in combination with surgery. Fifteen tumor-bearing dogs treated with T-LAK therapy combined with palliative resection of tumors were enrolled in the present study. T-LAK were generated from autologous peripheral blood mononuclear cells (PBMC) by culture with recombinant human interleukin -2 (rhIL-2) and solid phase anti-canine cluster of differentiation (CD)3 antibody. T-LAK were administrated intravenously at 2-4-week intervals. After the first administration of T-LAK, counts of PBL and T lymphocyte subsets (CD3(+), CD4(+) and CD8(+) cells) increased and the CD4/CD8 ratio decreased, with significant increases in CD8(+) cells (P

The emergence and global spread of extended-spectrum or AmpC β-lactamase (ESBL/AmpC)-producing Enterobacteriaceae in companion animals have led to the hypothesis that companion animals might be reservoirs for cross-species transmission because of their close contact with humans. However, current knowledge in this field is limited; therefore, the role of companion animals in cross-species transmission remains to be elucidated. Herein, we studied ESBL/AmpC-producing Enterobacteriaceae,Escherichia coliin particular, isolated from extraintestinal sites and feces of companion dogs. Whole-genome sequencing analysis revealed that (i) extraintestinalE.coliisolates were most closely related to those isolated from feces from the same dog, (ii) chromosomal sequences in the ST131/C1-M27 clade isolated from companion dogs were highly similar to those in the ST131/C1-M27 clade of human origin, (iii) certain plasmids, such as IncFII/pMLST F1:A2:B20/blaCTX-M-27, IncI1/pMLST16/blaCTX-M-15, or IncI1/blaCMY-2from dog-derivedE.coliisolates, shared high homology with those from several human-derived Enterobacteriaceae, (iv) chromosomalblaCTX-M-14was identified in the ST38 isolate from a companion dog, and (v) eight out of 14 tested ESBL/AmpC-producingE.coliisolates (i.e., ST131, ST68, ST405, and ST998) belonged to the human extraintestinal pathogenicE.coli(ExPEC) group. All of thebla-coding plasmids that were sequenced genome-wide were capable of horizontal transfer. These results suggest that companion dogs can spread ESBL/AmpC-producing ExPEC via their feces. Furthermore, at least some ESBL/AmpC-producing ExPECs andbla-coding plasmids can be transmitted between humans and companion dogs. Thus, companion dogs can act as an important reservoir for ESBL/AmpC-producingE.coliin the community.

Canine nasal carcinomas are often treated with radiotherapy. Presence of lysis of the cribriform plate by tumor invasion (stage 4 by modified Adams's staging system) is a well-known prognostic factor. In this study, dogs with stage 4 disease were divided into two subgroups based on the presence or absence of midline shift of the olfactory or frontal lobes of the brain (Stage 4a: without presence of midline shift. Stage 4b: with midline shift). The median survival time of dogs with midline shift was significantly shorter than that of dogs without midline shift (64 vs. 208 days). Our results indicate that the finding of a midline shift might have a prognostic significance in dogs with nasal carcinoma treated with radiotherapy.

The influence of transfusion of lymphokine-activated T killer cells (T-LAK) on inflammatory responses was examined in dogs after laparotomy. Plasma C-reactive protein (CRP) level, cell numbers of peripheral blood lymphocytes (PBLs) and T lymphocyte subsets (CD3(+), CD4(+) and CD8(+)) and mRNA expression levels of cytokines including interleukin (IL)-2, IL-12, IL-4, IL-10 and transforming growth factor (TGF)-β in peripheral blood mononuclear cells (PBMCs) were measured in dogs with (T-LAK group) or without (control group) a single T-LAK administration immediately after laparotomy. The plasma CRP level initially increased and then decreased to the normal range at 7 days after laparotomy in the T-LAK group, which was earlier than in the control group. The expression level of IL-10 mRNA showed a marked postoperative increase and was significantly higher than the preoperative level on day 7 (P

Prohibitin is an antiproliferative protein that is a product of a putative tumor suppressor gene. However, there is little information on prohibitins in companion animals. In this study, we cloned canine prohibitin mRNA using RT-PCR and 3′-RACE (Rapid Amplification of cDNA Ends). The sequence was well conserved compared with those of other mammals, including human. The deduced amino acid sequence translated from the open reading frame completely corresponded to the human sequence. Canine prohibitin mRNA was expressed in all normal mammary and tumor samples examined. These results suggest that this protein plays a vital role in cell growth mechanisms and may be related to the occurrence of canine mammary tumors.

The reactivity of different lectins with crude chromogranin A (CgA) obtained from different animals, namely, cow, horse, dog, pig, and dolphin, was examined to identify lectin(s) that would be useful as coating reagent(s) in a sandwich enzyme-linked immunosorbent assay (ELISA). Of the different lectins studied, the Amaranthus caudatus lectin (ACA), which is specific for the Thomsen–Friedenreich (T)-antigen (Galβ1-3GalNAc), was found to react with the CgA from different animals by western blotting. Purified rabbit anti-bovine CgA antibody was also found to cross-react with the crude CgA preparations. On the basis of these findings, a sandwich ELISA was developed with ACA as the coating reagent and anti-bovine CgA antibody as the probing antibody. Using this method, concentration-dependent curves ranging from 0.003 μg/mL to 25 μg/mL and from 0.02 μg/mL to 25 μg/mL were obtained for bovine CgA and canine CgA, respectively. Similarly, concentration-dependent curves were obtained for the equine, swine, and dolphin crude CgA extracts. Thus, ACA is concluded to be a valuable reagent for CgA detection in crude extracts from different animal species, and for CgA isolation/purification.

Human BRCA1 is familial breast cancer susceptibility gene. Recently, decreased BRCA1 mRNA and protein expression has been identified in sporadic breast tumors. In the reported human BRCA1 splicing variants, delta11b lacks the majority of exon11 and is suspected to have a distinct function in normal tissues. The splicing variants display a variety of expression pattern in breast cancer samples. Although mammary gland tumor is important disease in dog, there are few reports for BRCA1 in the canine tumors. In this study, we examined the relative amounts of BRCA1 splicing variants mRNA in canine normal and mammary tumor samples by RT-PCR to investigate whether there is the altered expression of variant mRNAs in the canine tumor as reported in human. The exon11b-defecting RT-PCR products were observed in all the normal tissues examined and the nucleotide sequence was quite similar to that of human BRCA1 delta11b. In some tumor samples, we did not detect the products targeted for exon10-13 and exon14-15, while these products were observed in all the normal samples examined. Especially, the relative amounts of the exon11-defecting products were remarkably decreased in most of the tumors (11/16).

application/pdf, Most anesthetics have an immuno-suppressive effect on cellular and neurohumoral immunity, and research shows that total intravenous anesthesia (TIVA) with propofol has a greater immuno-protective effect than inhalational anesthesia in human medicine. However, in veterinary clinics, these effects remain ambiguous. To clarify the details, we focused on propofol and isoflurane, investigating clinical blood hematology and immunological profiles drawn from healthy dogs under and after two anesthesia techniques. Twelve healthy adult beagles were included in this study, randomly assigned to the propofol anesthesia group (group P: n=6) or the isoflurane anesthesia group (group I: n=6). In both groups, the number of lymphocytes in peripheral blood decreased after 2 hr of anesthesia (2 hr), but group P showed significantly less decrease than group I. For T-lymphocyte subsets examined by flowcytometry, the ratio of CD3+, CD4+ and CD8+ lymphocytes in the peripheral blood mononuclear cell (PBMC) of group P at 2 hr also exhibited a high level compared to group I. Moreover, for mRNA expression of cytokines measured by real-time PCR, the IL2 (pro-inflammatory cytokine) of group P showed no decrease like group I. The IL10 (anti-inflammatory cytokine) of group P also showed no increase like group I, while both cytokines maintained nearly the same level until 2 hr. These results suggest that, compared to propofol, isoflurane had more strongly immuno-suppression caused by anesthesia, and propofol itself might have some immuno-protective effects. Thus, TIVA with propofol might benefit immunological support in the perioperative period of dogs.

Objective—To determine whether cross-reactivity exists between canine chromogranin A (CgA) and anti-human CgA antibody and investigate the usefulness of plasma CgA concentration measurements as an index of acute stress responses in dogs. Animals—12 healthy Beagles. Procedure—Canine CgA was extracted and purified from canine adrenal glands of cadaver dogs for studying cross-reactivity with anti-human CgA antibody. Western blotting with anti-human CgA antibody was performed. Blood samples were collected from dogs at 0, 10, 20, 30, 40, 60, 120, and 180 minutes after IV administration of saline (0.9% NaCl) solution or insulin. Canine plasma CgA concentrations were determined by use of a CgA ELISA kit with rabbit antiserum against the carboxy-terminal fragment of human CgA. Plasma cortisol and catecholamine (ie, norepinephrine and epinephrine) concentrations were measured by use of an ELISA and a high-performance liquid chromatography method, respectively. Results—Purified canine CgA was specifically detected by use of western blot analysis and an ELISA with anti-human CgA antibody. An increase in plasma CgA concentrations was observed in insulin-induced hypoglycemic dogs. Changes in plasma CgA concentration were correlated with changes in plasma cortisol or catecholamine concentrations of hypoglycemic dogs. Conclusions and Clinical Relevance—Use of the CgA ELISA kit for determination of human plasma CgA concentrations is applicable to the measurement of canine plasma CgA concentrations. Canine plasma CgA concentrations, along with measurements of plasma cortisol and catecholamine concentrations, correctly reflect insulin-induced hypoglycemic stressed conditions in dogs. Measurement of canine plasma CgA concentrations may provide a useful index for evaluation of an acute stress response. (Am J Vet Res 2005;66:1830–1835)

Renal effects of the selective alpha(2)-adrenoceptor agonist, medetomidine, were investigated in anesthetized dogs. Animals were administered medetomidine 20 and 40 microg/kg intravenously (IV) and 80 mug/kg intramuscularly (IM) or 1 ml of saline IV. Urine and blood samples were collected before and at 30, 60, 90 and 120 min following medetomidine injection. Mean arterial blood pressure (MABP), renal blood flow (RBF), glomerular filtration rate (GFR), urine volume (U(v)), urine osmolality (U(osm)), free water clearance (C(H2O)), fractional clearance of sodium (F(Na)), plasma osmolality (P(osm)), plasma glucose levels and plasma antidiuretic hormone (ADH) concentrations were measured. The results showed that IV administration of medetomidine initially increased MABP 5-15 min followed by long-lasting decrease. The initial hypertension was not observed after IM administration, which was accompanied by a more profound hypotensive effects. RBF, GFR, U(v), C(H2O) increased after IV injection and decreased after IM. Medetomidine increased FNa and Posm and decreased U(osm). Plasma glucose levels initially increased and subsequently decreased. Plasma ADH concentration was decreased by IV injection but increased by IM administration. Our data imply that: 1) IV administration of medetomidine at dose rates of 20 and 40 microg/kg results in profound diuresis up to 2 hr; 2) Suppression of ADH release from the CNS is one of the mechanisms of medetomidine-induced diuresis although it may not be the principal one.

ErbB subfamily genes, known as proto-oncogenes, encode receptor tyrosine kinases, and are expressed in relation to tumorigenesis of the mammary gland in humans. In this study, we examined the expression of erbB subfamily mRNAs in two canine normal mammary glands and 12 mammary tumor samples by reverse transcriptase-coupled polymerase chain reaction (RT-PCR). Each primer set was designed from the nucleotide sequence of the region conserved in erbB subfamily cDNA among other species. No erbB subfamily mRNAs were expressed in the normal mammary gland. In contrast, all of the subfamily mRNAs were expressed in a benign mammary tumor, and more than one type of the subfamily mRNA were observed in 11 malignant mammary tumors. The length of RT-PCR products were 380 bp for erbB1, 500 bp for erbB2, 644 bp for erbB3, and 416 bp for erbB4. These sequences were highly homologous to the cDNA sequences of other species. Therefore, these results suggest that the expression of erbB subfamily mRNAs in canine mammary tumors plays an important role in tumorigenesis of the mammary gland.

Based on the recent findings that show how recombinant human tumor necrosis factor (rh-TNF)-alpha has potent antitumor activity on human cancer patients when it locally administrated, we have tested the cytotoxicity of rh-TNF-alpha on 3 canine cultured cells: (1) canine kidney carcinoma (CKCa-1), (2) mastocytoma and (3) Mardin Darby canine kidney cells (MDCK). The cell surface expression of TNF-alpha receptors on these canine cells was also determined with anti-human TNF RI and RII polyclonal antibodies. Our data shows that on CKCa-1 which has TNF RI receptors rh-TNF-alpha induced cytotoxicity. By contrast, it exhibited no toxicity on canine mastocytoma which has mainly RII receptors. The data also suggest actinomycin D (ACT-D), an anticancer antibiotic, enhanced the cytotoxicity of rh-TNF-alpha. Combined with ACT-D, rh-TNF-alpha showed the cytotoxicity on MDCK which possessed both TNF RI and RII receptors. The results indicate that the cytotoxicity of rh-TNF-alpha depends on the presence of TNF RI receptors on canine tumor cells.

The mammary tumor is one of the popular neoplastic diseases in female dogs. In the present study, the expression of canine c-kit proto-oncogene in mammary tumor specimens was investigated to evaluate its potential usefulness as a tumor marker. By comparing the homology among the nucleotide sequences reported for human mouse, rat and feline c-kit c-DNA, a pair of primers was synthesized for the reverse transcriptase-polymerase chain reaction (RT-PCR) method. The RT-PCR product of canine spleen total RNA was shown to have 756 bp in size and to be highly homologous to the corresponding sequences reported for the mammalian species. The expression of c-kit transcript was detected in 11 mammary tumors of different histopathology including adenocarcinomas, benign and malignant mixed tumors. The level of the transcription in adenocarcinomas was significantly higher than those in malignant mixed tumors. Fifteen canine tumor specimens originated from various tissues were also tested for their c-kit transcript. In all of the mastocytoma samples examined, high expression of the mRNA was detected. Of other 12 tumors, only low level of RT-PCR products were detected in 5 samples, whereas no apparent amplification was observed in 7 tumors. These results indicate that the high expression of c-kit transcript is helpful for the diagnosis of canine mammary tumors.

β2-Microglobulin (β 2M ) forms the invariant chain of major histocompatibility complex (MHC) class I molecules and is essential for their structural stability and optimal functioning (Pedersen and others 1994). Loss or downregulation of MHC class I expression has been reported in various types of cancer (Chang and others 2003, Aptsiauri and others 2007). In human beings, mutations in β 2M are a significant mechanism for the total loss of MHC class I, and β 2M mutations have been described in colon carcinomas, melanomas and lymphomas. The mutational hot spot in β 2M mutations is exon 1 (Garrido and others 1997, Perez and others 1999), although no such mutations have been identified in breast cancers (Chen and others 1996). Several point mutations have been identified in β 2M in healthy domestic dogs, and the data have been deposited in the Broad Institute Dog Genome Project database (CanFam2.0—2,544,508 SNPs; http://www.broadinstitute.org/mammals/dog). However, almost all known canine point mutations remain uncharacterised (Chang and others 2007). To the best of our knowledge, no other group has studied β 2M mutations in canine mammary gland tumours. Therefore, the aim of the present study was to determine β 2M mutations in canine mammary gland simple and complex carcinomas. Approximately 1 g tissue samples were collected from the mammary gland tissues of seven healthy beagles, from the tumours of 11 dogs with mammary gland tumours (one Shih Tzu, two Labrador retrievers, two mixed breeds, one Welsh corgi, two toy poodles, one great Pyrenees, and two papillons), and from normal …

Erythroleukemia was observed in two unrelated cats infected with feline leukemia virus (FeLV) from the same household. Case 1, a 1-year-old neutered male cat developed erythroleukemia (M6) after a diagnosis of myelodysplastic syndrome (MDS-Er) on the criteria of FAB classification of acute leukemias. Case 2, a 1-year-old neutered female cat, which had close contact with Case 1, also developed erythroleukemia (M6Er). In both cases, marked proliferation of erythroid progenitor cells with disproportionally large numbers of immature forms was observed in the bone marrow. In Case 1, neoplastic proliferation of myeloid cells in the bone marrow was also noted at the terminal stage. Combination chemotherapy with daunomycin was partially effective for treatment of these erythroid neoplasias, but did not induce complete remission. Southern blot analysis using exogenous FeLV-specific probes indicated the clonal origin of these hematopoietic tumor cells. Furthermore, the erythroid and myeloid tumor cells in Case 1 were shown to be derived from independent transformed clones. A variant FeLV was shown to be integrated into the tumor cells in Case 1, while a full-length FeLV was found in both cases. Because these erythroid neoplastic diseases occurred in two unrelated cats kept in the same household and these diseases are rare, they may both have been associated with the same FeLV strain.

Changes of splenic lymphocyte subpopulation after Babesia microti and Babesia rodhaini inoculation in mice were examined by flow cytometric analysis. The B. microti inoculated mice showed a longer period of time from inoculation to the onset of increase or decrease parasitaemia (%), packed cell volume, total spleen cell numbers and surface immunoglobulin positive splenic cell numbers than respective periods in B. rodhaini inoculated mice. The Thy-1 positive cell numbers in B. microti inoculated mice and B. rodhaini inoculated mice pre-immunized with homologous parasites were significantly higher than that of B. rodhaini inoculated mice. The ratio of L3T4 positive cell/Lyt-2 positive cell after inoculation with B. microti was quite similar to that in B. rodhaini mice pre-immunized. However, the ratio in B. rodhaini inoculated mice revealed a lack of an increasing phase. These results suggested that the T-cell dependent early immune response, especially suppressor activity, was closely related to the difference in the course of infection between the non-lethal B. microti and the lethal B. rodhaini infection in mice.

A 9-month-old male Shih-Tzu dog had a right mandibular tumor composed of strands, or nest-like proliferation of epithelial cells with abundant fibrous stroma characterized by spheroid to large nodular deposition of amyloid with Congo-red stain. Globule calcification was also seen throughout the tumor tissue and the spheroid depositions often had a concentrically laminated structure (Liesegang rings). The case was diagnosed as amyloid-producing odontogenic tumor in a dog.

Dolphins in a captive environment are exposed to various kinds of stresses. Handling and transportation are stressful events for terrestrial mammals, and such stress may affect immune system function and increase susceptibility to infectious diseases. The same phenomenon could occur in dolphins, however, few studies have reported this in dolphins. The objective of this study was to evaluate the relationship between stress and polymorphonuclear (PMN) cell function of dolphins during transportation. Four bottlenose dolphins (Tursiops truncatus) were transported for 6 hr by truck. Serum cortisol levels, leukograms, phagocytosis, and superoxide production of PMN cells were evaluated during handling and transportation compared to resting values. The mean serum cortisol level was significantly increased during handling and transportation (p

A 19.5-year-old male mongrel dog developed a progressive lameness and swelling around the right carpus. A tumor (6 × 3 × 3 cm) was found in the caudal of distal antebrachium of the right forelimb, including tendons of the superficial digital flexor muscle and deep digital flexor muscle. No joint destruction was observed. The tumor consisted of round and spindle cells arranged in a compact sheet. There were occasional slit-like spaces or lumina, and areas rich in collagen fibers giving an appearance of tendon tissues. Neoplastic cells gave a positive immunoreaction to vimentin, but negative reactions to antibodies against S-100 protein, cytokeratin and myoglobin. Based on these findings, this tumor was diagnosed as a synovial sarcoma generating from the tendon and tendon sheath, which is very uncommon in dogs.

Bovine and canine chromogranin A were extracted and purified from each specie's adrenal glands. Isolated bovine 70 kDa protein showed 100% identity to bovine CgA reported previously, whereas isolated canine 68 kDa protein showed 83.3% identity to bovine CgA by the NH(2)-terminal amino acid sequence analysis. Rabbit antibody to purified bovine protein (CgA) was found to immunologically cross-reacted with purified canine protein (CgA). In sandwich ELISA with anti-bovine CgA, concentration-dependent curves were obtained ranging from 0.3 to 20 mug/ml for canine CgA. From these findings, sandwich ELISA with anti-bovine CgA is found to be useful to determine the concentration of canine CgA.

To clarify the role of nitric oxide (NO) in hemodialysis (HD)-related hypotension, the relationship between plasma NO metabolites (NOx) and blood pressure changes, and the effect of N(G)-monomethyl-L-arginine (L-NMMA), a NO synthase inhibitor, on changes in blood pressure were evaluated in an experimental renal dysfunctional dog model. In order to create a renal dysfunction model, gentamicin was administered to male beagles in which 7 of 8 renal artery branches had been ligated. Normal renal functional and dysfunctional dogs underwent 3 hr of HD per day for 3 days. HD induced a transient decrease in mean blood pressure in the normal renal functional dogs. In renal dysfunctional dogs, a continuous hypotension occurred with a gradual increase in the plasma NOx concentration during HD. Although L-NMMA prevented the fall in blood pressure, it did not significantly change the plasma NOx concentration during HD. These results suggest that NO contributes to HD-related hypotension in renal dysfunctional dogs but the plasma NOx concentration does not reflect the change in blood pressure.

Glucose-6-phosphate dehydrogenase (G6PD), malate dehydrogenase (MDH), and lactate dehydrogenase (LDH) activities and their isoenzyme patterns in B. rodhaini (BR) and B. microti (BM), the two major causative species of murine babesiosis, were examined. G6PD and LDH activities were higher in BR than those in BM, whereas MDH activity was lower in BR than that in BM. No differences were observed between BR and BM in the mobility of isoenzyme bands of G6PD and MDH. On LDH isoenzyme pattern, at least 5 bands were detected in BR, while only one band in BM. Since each subunit of LDH is known to be coded by different gene, these results suggests that BR and BM are able to be differentiated genetically.

A 12-year-old male cat with depression and dyspnoea was presented for investigation. Radiography and computed tomography revealed hydrothorax and solid masses involving the sternum, ribs and thoracic vertebrae. The cat died two days after first presentation, and postmortem examination revealed lung masses and proliferative bony lesions. Histologically, a neoplastic proliferation of epithelial cells was seen in the lungs, with a large amount of collagen and deposits of cholesterin. The bone lesions were also composed of neoplastic epithelial cells and abundant calcified osteoid, without atypia. A diagnosis of pulmonary adenocarcinoma with osteoblastic bone metastases was made. This is the first reported case of osteoblastic metastases in the cat.

An 11-year-old, mixed-breed dog with dyschezia, tenesmus and haematochezia was presented. A rectal stricture caused by an adenocarcinoma was diagnosed. Following the failure of a rectal 'pull-through' procedure, which dehisced seven days later, an incontinent end-on colostomy was performed, allowing amputation of the colorectum with the anus and perineal skin. A two-piece device consisting of a flange and a drainable pouch was used for postoperative faecal evacuation and collection. Mild peristomal dermatitis was the only complication. Patient management was easily carried out by the owner at home, and the dog survived for four months with a satisfactory quality of life. Incontinent end-on colostomy may prove to be a useful treatment for canine annular colorectal tumours.

The functions of polymorphonuclear cells (PMN) are the important non-specific defense mechanisms in the immune system. Especially marine mammals are protected by these mechanisms from the aquatic environment with a large variety of microorganisms. Therefore, we examined the PMN functions of bottlenose dolphins in order to obtain the normal ranges and to standardize the techniques. PMNs were isolated by using lymphocyte isolate solution whose density was 1.077; superoxide production was assessed by nitroblue tetrazolium reduction test (NBT) and phagocytosis was tested by using polystyrene latex beads. We showed that the optimal incubation time was 30 min in NBT assay and 12 hr in phagocytosis assay for dolphin PMNs.

Arthritis was produced in beagles by the immunological reaction to bovine serum albumin (BSA). Dogs immunized with BSA showed the development of delayed type-hypersensitivity response to BSA and the significant increase in the titer of serum anti-BSA antibodies. The development of arthritis and the increase in a number of nucleated cells in synovial fluid were observed by the injection with BSA into the knee joints of immunized dogs. The synovial membrane of BSA-injected joints revealed a remarkable villous hyperplasia of membrane, and an infiltration of lymphocytes and plasma cells around vessels, resulting in a lymphoid nodule-like formation. The depositions of IgG and C3 on the surface of the synovial membrane were also observed in BSA-injected joints. Histopathological and immunopathological findings indicated that the immune response to BSA in the knee joints could induce a rheumatoid arthritis like chronic synovitis in dogs.

To elucidate the role of T cell subpopulations in the protective cell-mediated immune response at the initial phase of infection with Babesia microti (BM) and B.rodhaini (BR), the changes in the course of infection and anti-parasite delayed type hypersensitivity (DTH) response after BM or BR inoculation were investigated in Lyt-2+ T cell- or L3T4+ T cell-depleted mice. Depletion of Lyt-2+ T cells strongly enhanced the resistance to BM infection, whereas it increased the susceptibility to BR infection. In contrast, depletion of L3T4+ T cells increased susceptibility to BM infection, while it enhanced resistance to BR infection. The anti-parasite DTH response in BM-infected mice was significantly enhanced by depletion of Lyt-2+ T cells, while significantly reduced by depletion of L3T4+ T cells. No effects of depletion of either Lyt-2+ or L3T4+ T cells on DTH response was observed in BR-infected mice. From these results, it was suggested that the roles of Lyt-2+ and L3T4+ T cells in the protective cell-mediated immune response at the initial phase of infection were different between BM- and BR-infected mice, resulting in the difference in their course of infection.

In vitro cultivation of Babesia rodhaini (BR) and Babesia microti (BM) was attempted. When RPMI1640 was supplemented with 30 or 40% of non-treated fetal bovine serum (FBS), the gas mixture of 3% CO2-8% O2 best supported the growth of both parasites. Under this optimized condition, the percent parasitized erythrocytes peaked to approximately 4- and 2-times initial values for Br and BM, respectively. The cultivated parasites retained the infectivity to the host mice. BM showed the characteristic feature of division during cultivation. However, the lots of FBS will have to be taken into consideration, since the FBS lots were shown to give large varieties to the results. Selection of the appropriate FBS lot may yield the better growth of these protozoa.

A comparative study was carried out on the glucose metabolism in Babesia microti (BM) and Babesia rodhaini (BR) by analyzing the enzyme activities. The lactate dehydrogenase (LDH) activity in BM showed significantly lower values than that in BR, whereas citrate synthase (CS) and malate dehydrogenase (MDH) activities were remarkably higher in BM. In addition, pyruvate dehydrogenase (PDH), isocitrate dehydrogenase (ICDH), alpha-ketoglutarate dehydrogenase (KGDH), and succinate dehydrogenase (SDH) activities also tended to be higher in BM. Then, the change of enzyme activities related to the proliferation of parasites was examined. In BM infected mice, the parasitemia increased from day 15 to day 19 after inoculation (a.i.). While BM showed decrease of G6PD and LDH activities at day 19 a.i., it showed remarkably increased activities in CS and MDH (368 and 8,842 nmol/min.mg protein, respectively). In addition, PDH, ICDH, KGDH, and SDH activities also tended to increase from day 15 to 19 a.i. In BR infected mice, parasitemia increased from day 9 to day 12 a.i. LDH activity showed a considerable increase at day 12 a.i. (12,920 IU/mg.protein). Although CS and MDH activities also showed a slight increase at day 12 a.i., the activities of PDH, ICDH, KGDH and SDH didn't change from day 9 to 12 a.i. Since these changes observed in the enzyme activities of BM and BR seemed to be correlated with their proliferation, it was suggested that BM and BR depended on aerobic and anaerobic pathways, respectively, for their glucose metabolism.

A 7-year-old male Yorkshire terrier was examined for multiple plaques and nodules on the skin. The clinical, cytological and histopathological features indicated a malignant histiocytosis. Cytoreductive chemotherapy produced moderate clinical improvement, but died at day 90 after the first admission. Pathological examination revealed the neoplastic histiocytes in the skin as well as in the myocardium and skeletal muscles.

Plasma iothalamate (IOT) disappearance rates were measured after a single-injection of IOT (113.8 mg/kg, IV) in cats with experimentally induced renal failure. The disappearance rates especially fitted into the one compartment model. The mean value of plasma disappearance rates of IOT in these cats with induced renal failure (2.16 +/- 0.240 x 10(-3) micrograms/ml/min) was markedly lower than that of clinically healthy cats (4.10 +/- 1.00 x 10(-3) micrograms/ml/min). These results demonstrate that IOT clearance is available for evaluation of renal function in cats.

Plasma methylguanidine (MG) and creatinine (CRN) concentrations were measured in 11 cats with experimentally induced acute renal failure by a two-stage surgical procedure. According to the progression of renal failure, both plasma MG and CRN levels increased. A significant positive correlation (y = 0.187X - 0.379, lambda = 0.9176, P0.001) was found between plasma MG and CRN levels. These results suggested that the increase in plasma MG level was an available indicator for uremic status in cats.