Glomus tumor of the liver in a cow.

An 11-year-old Holstein-Friesian cow exhibited anorexia and jaundice. A large mass was found in the liver during necropsy. Macroscopically, the mass was composed of dark red multilobular tissue and a centrally located abscess, which was connected to the hepatic duct. Histologically, the mass consisted of proliferation of small neoplastic cells and was demarcated from the hepatic parenchyma by a thick region of granulation tissue. The neoplastic cells were predominantly arranged in solid sheets, but they also formed blood-filled cancellous structures, and proliferating foci were seen around blood vessels. Periodic acid-Schiff reaction demonstrated that a fine basement membrane-like structure surrounded the neoplastic cells. Immunohistochemically, the neoplastic cells were positive for vimentin and alpha smooth muscle actin and negative for cytokeratin, factor VIII-related antigen, chromogranin and desmin. Based on its histopathological features, the hepatic neoplasm was diagnosed as a primary glomus tumor. This is the first report about a primary glomus tumor of the liver in a cow.

Humeral chondrosarcoma in a Hokkaido brown bear (Ursus arctos yesoensis).

Humeral chondrosarcoma was found in an 18-year-old male Hokkaido brown bear (Ursus arctos yesoensis). Necropsy revealed a large firm mass under the left superficial pectoral muscle at the axillary region. The mass involved the left shoulder joint and peripheral muscles, and connected to the head of the humerus with osteolysis. Histopathologically, the mass was composed of irregularly shaped myxomatous to cartilaginous tumor lobules. The tumor cell showed moderate nuclear atypia with a relatively high mitotic index, especially in the edges of the myxomatous lobules. The tumor cells were positively immunostained with vimentin and S-100 protein. Based on these findings, the tumors were diagnosed as chondrosarcoma. Metastases were found in the left axillary lymph node, lungs, liver and kidney.

Atypical AA amyloid deposits in bovine AA amyloidosis.

In bovine amyloid protein A (AA) amyloidosis, amyloid deposits are typically observed in the kidney and spleen at necropsy. To determine the distribution of amyloid deposits in cows affected with AA amyloidosis, we examined organs known to be sites of amyloid deposits that are also processed for human consumption in 14 cows: 11 with typical clinical symptoms (typical amyloidosis) and three with no typical clinical symptoms (atypical amyloidosis). We found unusually high amounts of amyloid deposits in the tongue and other organs in all 14 cows regardless of the presence or absence of clinical amyloidosis symptoms. Cows with typical amyloidosis had heavier amyloid deposits in the spleen and renal glomeruli than cows with atypical amyloidosis. From clinical symptoms and histological examinations, we found that cows with typical and atypical amyloidosis can be classified into two groups, class I and class II, according to the presence or absence of heavy amyloid deposits in the spleen and renal glomeruli. However, no significant differences were observed between the amyloid fibrils of class I and class II amyloidosis by electron microscopy and Western blot analysis.

Pathogenesis of experimental amyloid protein A amyloidosis in sore hocks-affected rabbits.

Although the experimental transmission of amyloid protein A (AA) amyloidosis with amyloid-enhancing factor has been studied intensively, its pathogenesis remains obscure. We previously found that rabbits affected with 'sore hocks' (SH) uniquely developed AA amyloidosis in response to primary inflammatory stimulation followed by the administration of bovine AA fibrils. However, it is unknown why only the rabbits with preexisting SH developed experimental AA amyloidosis. There may be hidden factors in the SH status that stimulate the mechanism of cross-species transmission of AA amyloidosis. To examine the essential factors in the development of experimental AA amyloidosis in SH-affected rabbits, we studied the etiology of SH in rabbits pathologically and bacteriologically. In addition, we developed artificial SH symptoms in normal rabbits by use of an adjuvant prepared from Staphylococcus aureus (StA) isolated from a spontaneous SH-affected rabbit, and we evaluated the incidence of AA amyloidosis in rabbits with or without experimental SH symptoms. We found that StA administration was extremely efficient at stimulating the induction of experimental AA amyloidosis, and the influence of SH was required. We found that the persistent S. aureus infection in SH facilitates the development of experimental AA amyloidosis in rabbits and that the inflammatory stimulation provided by SH acts as an additional accelerator in experimental AA amyloidosis.

New findings concerning eosinophilic substance deposition in mouse nasal septum: sex difference and no increase in seniles.

An eosinophilic substance (ES) is usually observed in the mouse nasal septum. In contrast to textbooks and one report describing ES as amyloid, a previous study by the authors revealed that ES is not amyloid but consists of collagen and an amorphous material. Furthermore, it was suggested that the amorphous material was produced by clear HE-stained nasal gland epithelial cells present at the dorsal portion directly above the vomeronasal organ. In this histological examination, ES deposition showed sex difference (more intense in males than in females). ES increased with age but not in seniles, suggesting that the increase has a limit. In the detailed examination using subserial HE-stained nasal sections, it was revealed that the clear HE-stained nasal glands continued to the vomeronasal glands, which communicated with the lumen of the vomeronasal organ, and the vomeronasal gland epithelial cells contained strongly periodic acid-Schiff (PAS) positive granules, similar to the clear HE-stained nasal gland epithelial cells. ES also deposited in the interstitium of the vomeronasal glands. The results suggested a possibility that ES deposition may be related to vomeronasal organ.

Prevention of Escherichia coli O157:H7 infection in gnotobiotic mice associated with Bifidobacterium strains.

Previous reports have shown that Escherichia coli O157:H7 infection is strongly modified by intestinal microbes. In this paper, we examined whether bifidobacteria protect against E. coli O157:H7 infections using gnotobiotic mice di-associated with Bifidobacterium strains (6 species, 9 strains) and E. coli O157:H7. Seven days after oral administration of each Bifidobacterium strain, the mice were orally infected with E. coli O157:H7 and their mortality was examined. Bifidobacterium longum subsp. infantis 157F-4-1 (B. infantis 157F) and B. longum subsp. longum NCC2705 (B. longum NS) protected against the lethal infection, while mice associated with all other Bifidobacterium strains, including type strains of B. longum subsp. infantis and B. longum subsp. longum, died. There were no significant differences in the numbers of E. coli O157:H7 in the faeces among the Bifidobacterium-associated mouse groups. However, the Shiga toxin concentrations in the cecal contents and sera of the GB mice associated with B. infantis 157F and B. longum NS were significantly lower than those of the other groups. However, there were no significant differences in the volatile fatty acid concentrations and histopathological lesions between these two groups. These data suggest that some strains of B. longum subsp. longum/infantis can protect against the lethal infections of E. coli O157:H7 by preventing Shiga toxin production in the cecum and/or Shiga toxin transfer from the intestinal lumen to the bloodstream.

Isolation and characterization of monoclonal antibodies against bovine serum amyloid A1 protein.

Bovine AA amyloidosis is the most frequently encountered amyloid type in cattle, and it is characterized by an extracellular deposition of pathological amyloid A (AA) protein. Because of the lack of a specific monoclonal antibody (mAbs) against bovine amyloid A (bAA) protein and its precursor, bovine serum amyloid A1 (bSAA1), at present anti-bAA rabbit antiserum and anti-human AA or SAA mAbs are widely used for diagnosis and analysis of bovine AA amyloidosis. In this study, three specific mAbs against bSAA1 were isolated by immunization using synthetic peptides of bSAA1, and these mAbs showed higher detection ability and specificity to bAA and bSAA1 than rabbit antiserum and anti-human AA or SAA mAbs in Western blot analysis and immunohistochemistry. These novel mAbs will be valuable in the development of a more precise immunochemical diagnostic tool for bovine AA amyloidosis, as well as for studying the pathophysiological mechanisms involved in this disease.

Detection of serum amyloid A isoforms in cattle.

Two-dimensional electrophoresis and immunoblotting were used to characterize 7 serum amyloid A (SAA) isoforms in cows with amyloidosis or chronic inflammation. Five SAA isoforms (isoelectric point [pI] 5.6, 6.1, 6.4, 6.8, and 7.7) were detected in all 10 amyloidosis serum samples, while 2 isoforms, pI 5.2 and 8.6, were detected in 4 and 9 of the samples, respectively. The same 7 isoforms were also detected in the serum of cows with chronic inflammation, but SAA pI 5.2 and 8.6 were detected in only 1 and 2 of 10 samples, respectively. It was concluded that although an amyloid-specific SAA isoform was not detected, examination of SAA isoform detection patterns may help identify bovine amyloidosis.

Deposition process of eosinophilic substance in mouse nasal septum.

An eosinophilic substance is usually observed in the mouse nasal septum, and its volume increases with age. In contrast to descriptions in textbooks defining the eosinophilic substance as amyloid, our previous report revealed that the observed eosinophilic substance is not amyloid, but consisted of collagen and an amorphous material. Furthermore, it was suggested that the amorphous material was produced by the clear hematoxylin and eosin (HE)-stained nasal gland epithelial cells. In this study, we investigated the deposition process of the amorphous material produced by nasal gland epithelial cells in the interstitium morphologically. In most cases, the amorphous materials in the clear HE-stained nasal gland epithelial cells accumulated at the basal portion. Collagen fibers surrounding the nasal glands partially disappeared, whereas the amorphous material in contact with the rough endoplasmic reticulum of the nasal gland epithelial cells continued to the amorphous material in the interstitium. These findings suggested that the amorphous material produced by the clear HE-stained nasal gland epithelial cells migrated to the interstitium through the partial opening of the basement membrane.

Nephrotic syndrome induced by dibasic sodium phosphate injections for twenty-eight days in rats.

Sprague-Dawley rats received once daily tail-vein injections of 360 mM dibasic sodium phosphate solution at 8 mL/kg for fourteen or twenty-eight days. Clinical examination revealed persistent proteinuria from three days after the first dosing and thereafter severe proteinuria from eight days or later in the phosphate-treated groups. Proteinuria developed without remission even after fourteen-day withdrawal in the fourteen-day dosed group. Phosphate-treated animals developed lipemia, hypercholesterolemia, anemia, higher serum fibrinogen levels, and lower serum albumin/globulin ratios on day 29. Renal weight increased significantly compared with control animals, and the kidneys appeared pale and enlarged with a rough surface. Histopathologically, glomerular changes consisted of mineralization in whole glomeruli, glomerular capillary dilatation, partial adhesion of glomerular tufts to Bowman's capsule, and mesangiolysis. Ultrastructural lesions such as an increased number of microvilli, effacement of foot processes, and thickening of the glomerular basement membrane, and immunocytochemical changes in podocytes, mainly decreased podoplanin-positive cells and increased desmin expression, were also conspicuous in the phosphate-treated rats for twenty-eight days. Marked tubulointerstitial lesions were tubular regeneration and dilatation, protein casts, mineralization in the basement membrane, focal interstitial inflammation, and fibrosis in the cortex. These clinical and morphological changes were similar to features of human nephrotic syndrome.

Brain abscess in a Japanese black calf: utility of computed tomography (CT).

Computed tomography (CT) was used for diagnosis of brain abscess in a 6-month-old, Japanese black calf presented with neurological dysfunction, compulsive circling and vision disturbance. CT images showed asymmetric lateral ventricles, and presence of intra-cranial multiple low absorption lesions surrounded by capsule suggestive of abscess in the right cerebral hemisphere. Postmortem examination revealed marked swelling of right cerebral hemisphere and olfactory bulb. Multilocular large abscess containing creamy pus was found to occupy most area of periventricular and lateral ventricle. Fusobacterium necrophrum was isolated from the abscess contents as the causative agent. These results demonstrate that CT is useful tool for tentative diagnosis of bovine brain abscess.

Experimental induction of amyloidosis by bovine amyloid fibrils in Sore Hock rabbits.

We report the experimental amyloidosis associated with administration of bovine amyloid fibrils in rabbits afflicted by Sore Hock (SH), which is ulcerative pododermatitis. Two groups of SH-afflicted rabbits were subjected to five inflammatory stimulations at intervals of 4 days by intraepithelial injection of a mixture consisting of Freund's complete adjuvant and lipopolysaccharide. One group of rabbits was administered amyloid in conjunction with the last inflammatory stimulation and the other group was not. For additional control, two groups were designed. A third group consisted of rabbits without SH, which were subjected to five stimulations and were administered amyloid. A fourth group consisted of SH-afflicted rabbits, subjected to 0-4 stimulations and administered amyloid. Amyloid depositions were observed in SH-afflicted rabbits, which had been stimulated five times and given amyloid (18/18). In the 4th group, only one rabbit, which had been subjected to four stimulations, showed amyloid depositions. No amyloid depositions were observed in the other rabbits. These results suggest that bovine AA amyloid fibrils have an amyloid-enhancing factor-like effect on SH-afflicted rabbits.

Pancytopenia with bleeding tendency associated with bone marrow aplasia in a Holstein calf.

An 11-day-old Holstein calf presented with a high rectal temperature and tachypnea. Treatment with antibiotics and non-steroidal anti-inflammatory drugs did not improve the clinical signs. Bleeding tendency, with several hemorrhage spots on the body surface, appeared five days after admission. Severe pancytopenia was observed in the blood examination. The calf died on the 11th day after admission with severe bleeding from an injection site. Necropsy findings revealed that the pancytopenia had resulted from severe bone marrow aplasia. A congenital disorder was suspected to be the cause of pancytopenia associated with bone marrow aplasia.

Immunohistochemical distribution of amyloid deposits in 25 cows diagnosed with systemic AA amyloidosis.

The distribution of amyloid deposits was histopathologically and immunohistochemically examined in 25 cows aged 5 to 10 years that had been diagnosed with systemic AA amyloidosis. This examination revealed that amyloid deposits were also present in the hypophysis, ovary, uterus, mammary gland and skeletal muscle, in addition to the liver, kidney, spleen, pancreas, thyroid gland, adrenal gland, gastrointestinal mucosa, heart, lung and lymph nodes. The examined cows tended to have chronic inflammations, including chronic mastitis (six cases) or chronic pneumonia (four cases), which is thought of as a causative agent of AA amyloidosis. In contrast, five cases did not exhibit any chronic inflammation.

Unexpectedly high incidence of visceral AA-amyloidosis in slaughtered cattle in Japan.

Experimental mouse AA amyloidosis can be transmissible by dietary ingestion of amyloid fibrils and it is well known that AA amyloidosis occasionally develops in aged cattle. Bovine liver and intestine have conventionally been used in Oriental foods, and the incidence of visceral AA amyloidosis in slaughtered cattle was evaluated. Renal tissues from 302 aged cattle older than 4 years were obtained from a local abattoir. Amyloid deposition was microscopically examined and amyloid protein was immunochemically determined. Renal amyloid deposition was seen in 15 out of 302 cattle with no previous history of diseas, an incidence of 5.0%. Amyloid protein in these cattle was AA and they had pathological findings in their visceral organs on gross examination. The incidence of visceral AA amyloidosis in slaughtered cattle in this study was disturbingly high compared with those (0.4-2.7%) previously reported from Japan and other foreign countries. AA amyloidosis is a life-threatening complication in patients with chronic inflammatory diseases and these patients at risk should avoid ingesting food that may possibly contain AA amyloid fibrils. More detailed information on cattle amyloidosis is required to guarantee the safety of our food.

Possible enhancing effects of atrazine on growth of 7,12-dimethylbenz(a) anthracene-induced mammary tumors in ovariectomized Sprague-Dawley rats.

Atrazine, one of the most commonly used herbicides in the world, has been reported to have endocrine disrupting effects in vivo. In the present experiment, influence of dietary atrazine on the late promotion/progression stage of mammary carcinogenesis in ovariectomized female Sprague-Dawley rats was examined after a single intragastric administration of 7,12-dimethylbenz(a)anthracene (DMBA). When the incidence of palpable mammary tumors reached about 50%, the animals were subjected to ovariectomy and divided into tumor bearing [DMBA-Tumor(+)] and non-tumor bearing [DMBA-Tumor(-)] groups, with subgroups of each fed a soybean-free diet containing 0, 5, 50, or 500 p.p.m. atrazine for 34 weeks. At the completion of the study, the tumor volume in the 50 and 500 p.p.m. treatment Tumor(+) subgroups was greater than in the 0 p.p.m. control case. In the DMBA-Tumor(-) group, higher incidences and volumes of the mammary tumors, with or without statistical significance (P <0.05), were observed in the 50 and 500 p.p.m. subgroups. Atrazine treatment tended to increase proportion of estrogen receptor alpha-positive tumors and stimulated cell proliferation in the DMBA-Tumor(+) group, but with no clear effects on serum hormone levels. The present study indicates that atrazine has a potential for enhancing the growth of mammary tumors, partly through increasing cell proliferation in the promotion/progression stage in female rats under ovarian hormone-free conditions.

Neuropathological observation of rabbits (Oryctolagus cuniculus) affected with raccoon roundworm (Baylisascaris procyonis) larva migrans in Japan.

Larvae of the raccoon roundworm, Baylisascaris procyonis (B. procyonis) are a known cause of cerebrospinal larva migrans in animals and humans. The present paper described details of the central nervous lesion in the rabbits (Oryctolagus cuniculus) affected with B. procyonis larva migrans in Japan. Clinically affected animals showed neurological signs including circling, torticollis, tremor of head, or ataxic gait. The most characteristic pathological alterations were large malacic lesions associated with an activated astroglial proliferation which was seen at the corpus medullare in the cerebellum including the cerebellar peduncle. Moreover, focal malacic lesions with perivascular cuffing and infiltration by lymphocytes and heterophiles were scattered everywhere throughout the brain. In these lesions or normal-appearing areas away from obvious lesions, ascarid larvae, about a maximum 65-75 micro m in diameter, were recognized. Other prominent features were minute lesions (we call them migration tract-like lesions) composed of lymphocytes, hemosiderin-laden macrophages and reactive astrocytes scattering throughout the cerebrum. In this study, we demonstrated ascarid larvae in only eight out of 23 animals diagnosed as B. procyonis larva migrans. Since it is not always possible to detect the larvae, the possibility of B. procyonis larva migrans must be given serious consideration to the characteristic lesions described above.

Lack of significant effects of genistein on the progression of 7,12-dimethylbenz(a)anthracene-induced mammary tumors in ovariectomized Sprague-Dawley rats.

Genistein is thought to be one of the possible factors for decreasing the incidence of breast cancer in Asian peoples who take soy-rich diets. However, some experimental data suggest that genistein can stimulate breast cancer development via its estrogenic activities. To clarify the influence of genistein on the promotion/progression stage of mammary carcinogenesis, female Sprague-Dawley rats received a single intragastric administration of 7,12-dimethylbenz(a)anthracene (DMBA). When the incidence of palpable mammary tumors reached about 50%, all animals were then subjected to ovariectomy and divided into tumor-bearing [DMBA-tumor (+)] and no-tumor-bearing [DMBA-tumor (-)] groups, with subgroups of each treated with genistein at concentrations of 0, 25, or 250 ppm in soybean-free diet for 36 wk. At terminal sacrifice, the 25-ppm subgroup of DMBA-tumor(+) had a higher tumor incidence and volume, whereas the 250-ppm subgroup showed lower incidence, number, and volume than the 0-ppm subgroup, although differences were not statistically significant. In the DMBA-tumor(-) groups, eventual tumor volumes in the genistein-treated groups were dose dependently smaller than in the 0-ppm subgroup, although again without statistical significance. The present study indicates that genistein does not exert clear inhibitory effects on mammary carcinogenesis in the promotion/progression stage in female rats under ovarian hormone-free conditions.

Enhancing effects of beta-estradiol 3-benzoate but not methoxychlor on the promotion/progression stage of chemically-induced mammary carcinogenesis in ovariectomized rats.

Modifying effects of beta-estradiol 3-benzoate (EB) and methoxychlor (MXC), a pesticide which possesses weak estrogenic activity, on 7,12-dimethylbenz(a)anthracene (DMBA)-induced mammary carcinogenesis were investigated in ovariectomized or intact female Sprague-Dawley rats. Twenty-eight weeks after a single DMBA (100 mg / kg body weight) initiation, when the incidence of mammary tumor-bearing rats had reached 75%, a number of the animals were subjected to ovariectomy in order to obtain 3 groups: i) tumor-bearing, ovariectomized group; ii) tumor-bearing, intact group; iii) no-tumor, ovariectomized group. Subsequently animals of each group were subjected to subcutaneous implantation of 0.5 mg EB or given diet containing 1000 ppm MXC for 13 weeks. Although the incidences, multiplicities and volumes of the palpable tumors gradually decreased after ovariectomy, EB treatment stimulated tumor growth in the tumor-bearing, ovariectomized group thereafter. A similar effect of EB treatment was also observed in the no-tumor, ovariectomized group. However, MXC did not show any effect in the tumor-bearing, or no-tumor ovariectomized groups, except that the multiplicity of tumors was significantly decreased by MXC treatment in the tumor-bearing, intact group. The results of our study suggest that MXC has no promotion / progression effect, but rather possesses a weak inhibitory effect, whereas the strongly estrogenic substance EB clearly enhanced DMBA-induced mammary tumorigenesis.