Ptaquiloside and Pterosin B Levels in Mature Green Fronds and Sprouts of Pteridium arachnoideum.

Pteridium arachnoideum, a fern of the Pteridium aquilinum species complex found in South America, is responsible for several different syndromes of poisoning. Cases of bovine enzootic hematuria and upper alimentary squamous cell carcinoma are both frequent occurrences in Brazil, whereas only bovine enzootic hematuria is noted with any frequency around the world. The reason for the high frequency of upper alimentary squamous cell carcinoma in Brazil is not currently known. One possible explanation may be the higher levels of ptaquiloside and pterosin B in Brazilian Pteridium than those present in the plant in other countries. However, these levels have not yet been determined in P. arachnoideum. Thus, the present study aimed to measure and compare ptaquiloside and pterosin B levels in mature green fronds and sprouts of P. arachnoideum collected from different locations in Brazil. Samples of P. arachnoideum were collected from the states of Minas Gerais and Rio Grande do Sul. A total of 28 mature leaf samples and 23 sprout samples were used. The mean concentrations of ptaquiloside and pterosin B present in the mature green fronds of P. arachnoideum ranged from 2.49 to 2.75 mg/g and 0.68 to 0.88 mg/g, respectively; in P. arachnoideum sprouts, mean concentrations of ptaquiloside and pterosin B ranged from 12.47 to 18.81 mg/g, and 4.03 to 10.42 mg/g for ptaquiloside and pterosin B, respectively. Thus, ptaquiloside and pterosin B levels in P. arachnoideum samples collected in Brazil were higher in sprouts than in mature green fronds, as observed in other countries. However, there was no variation in ptaquiloside levels among plants collected from different cities in Brazil. The high frequency of upper alimentary squamous cell carcinoma in Brazilian cattle may not be attributed to greater levels of ptaquiloside and pterosin B in P. arachnoideum than in other Pteridium species in other countries.

Bovine upper alimentary squamous cell carcinoma associated with bracken fern poisoning: Clinical-pathological aspects and etiopathogenesis of 100 cases.

Upper digestive tract (UDT) cancer is rare in cattle, however in Southern Brazil, the UDT squamous cell carcinomas (SCCs) are relatively common and have been associated with bracken fern consumption and the presence of papillomas. Although a theory of pathogenesis considers bovine papillomavirus type 4 (BPV-4) as a cofactor in the development of these SCCs, some aspects of the etiopathogenesis of this disease need to be more investigated. In fact, detection of BPV-4 in UDT papillomas is scarce in other regions of the world and has not been performed in Brazil. Therefore, this study had two aims: 1) to analyze the epidemiological, clinical and pathological aspects of 100 natural cases of SCCs in the UDT of cattle grazing on bracken fern (Pteridium arachnoideum) highly contaminated areas, investigating the associations between these parameters; and 2) to investigate the presence of papillomavirus DNA by polymerase chain reaction (PCR) in the UDT papillomas (n = 47) from 30 cattle that also had UDT SCCs. There were statistically significant associations between clinical signs and tumor localization in the UDT; between histological grade of differentiation and tumor localization; and a trend towards significant association between histological grade of differentiation and presence of metastases. The average age of cattle with oropharyngeal SCCs was 7.39 years, with statistically significant difference comparing to cattle with esophageal SCCs (8.6 years). No statistical association was observed among other clinical-pathological parameters (growth pattern and primary site of the tumor) analyzed. No BPV DNA was detected in papillomas by PCR. Therefore, these results suggest the possibility that papillomas of the UDT are not necessarily associated with BPV infection.

Cyclooxygenase-2 immunoexpression in intestinal epithelium and lamina propria of cats with inflammatory bowel disease and low grade alimentary lymphoma.

BACKGROUND: Cyclooxygenase 2 (COX-2) is an inducible isoform by cellular activation, proinflammatory cytokines and growth factors. The aims of the current study were to evaluate COX-2 immunoexpression in epithelial and lamina propria (LP) of cats with inflammatory bowel disease (IBD) and low grade alimentary lymphoma (LGAL), as well as to correlate them with clinical signs and histopathological scoring. Cats diagnosed with IBD and LGAL (2007-2013) were included in the current study. Feline chronic enteropathy activity index (FCEAI) was calculated for all cases. Control group was composed by 3 healthy indoor cats and 5 sick cats died or were euthanized (non-gastrointestinal illness). Diagnosis and classification of IBD and LGAL was established according to the WSAVA gastrointestinal standardization group template and the National Cancer Institute formulation, respectively. Furthermore, a modified WSAVA template was applied for LGAL evaluation. Immunolabelling for COX-2 (polyclonal rabbit anti-murine antibody) was performed on biopsy samples. Epithelial and LP (inflammatory or neoplastic cells) COX-2 immunolabelling was calculated according to the grade and intensity. The most representative segment scored by the WSAVA and the modified WSAVA were used for statistical analysis. RESULTS: Significant difference was found regarding COX-2 intensity overexpression in the epithelial cells of IBD and LGAL groups when compared to control cats, but not between the groups of sick cats, whereas no differences were found regarding the grade of immunoreactivity between groups. No difference was found for COX-2 immunoexpression at the LP between all groups. However, 3 cats from LGAL group showed COX-2 expression in neoplastic cells at the LP. There were no correlations between epithelial or LP COX-2 expression and FCEAI and histological alterations. CONCLUSIONS: Increased COX-2 intensity at the epithelial cells observed in cats with IBD and LGAL may be secondary to the inflammatory response or a protective function in the intestinal reparation. COX-2 expression at the LP was presented in 33% of LGAL. This result provides a reason for further investigation concerning the role of COX-2 expression in feline alimentary lymphoma.

Leiomiossarcoma gástrico associado a hérnia de hiato em cão da raça Shar-pei: relato de caso / Gastric leiomyosarcoma associated with hiatal hernia in a Shar-pei: a case report

Os tumores do sistema digestório não são comuns na medicina veterinária e as neoplasias de estômago representam menos de 1% de todos os tumores malignos. O leiomiossarcoma é o segundo tumor que mais acomete o trato gastrointestinal de cães, com predisposição para as regiões de jejuno e ceco, mas qualquer porção pode ser afetada, desde o esôfago até o reto. O presente trabalho relata um caso em um canino da raça Shar-pei, macho, com 13 anos de idade e histórico de episódios eméticos pós- prandiais e emagrecimento progressivo havia oito meses. Após avaliação radiográfica e endoscópica, foi indicada intervenção cirúrgica e observada tumoração gástrica associada a hérnia de hiato. O diagnóstico de leiomiossarcoma gástrico foi estabelecido por meio de análise histopatológica. O paciente apresentou pouco tempo de sobrevida, entretanto destaca-se a raridade desse tipo de neoplasia gástrica e da associação desse tipo tumoral com hérnias de hiato, assim como a importância da análise histopatológica para o correto diagnóstico desse tipo de patologia.(AU)

Gastrointestinal system tumors are not common in Veterinary Medicine and stomach neoplasia represents less than 1% of all malignant tumors. Among digestive neoplasias, the leiomyosarcoma is the second most common with predisposition to the regions of the jejunum and cecum, but any portion from the esophagus to rectum can be potentially affected. This is a case report of a 13-year-old male Shar-pei dog with medical history of post prandial emetic episodes and progressive weight loss lasting eight months. Surgical intervention was indicated after radiographs and endoscopic evaluation, where a gastric mass and hiatal hernia were found. The diagnosis of gastric leiomyosarcoma was established only after histopathological analysis. Despite the patient having little survival time, this clinical presentation of a gastric tumor associated with hiatal hernia is rare, and exalts the importance of histopathological studies as a tool for the correct diagnosis in these unusual diseases.(AU)

Is Serum Total LDH Evaluation Able to Differentiate between Alimentary Lymphoma and Inflammatory Bowel Disease in a Real World Clinical Setting?

CONTEXT: An increase in enzyme lactate dehydrogenase (LDH) in serum is a negative prognostic factor for survival in cats affected by lymphoma. Measuring LDH at the time of diagnosis has been studied for differentiating neoplastic disease from non-neoplastic disease in dogs. Inflammatory bowel disease (IBD) and alimentary lymphoma are common diseases in cats. OBJECTIVE: The aim of this study was to determine whether elevation of total LDH occurred in cats with alimentary lymphoma and non-neoplastic gastrointestinal disease, such as IBD, and to evaluate whether this enzyme is useful in supporting the differential diagnosis of these specific diseases. MATERIALS AND METHODS: A prospective non-randomized controlled study was carried-out in a real world setting of three Italian private veterinary clinics. Seventy-one client-owned cats with a history of chronic gastrointestinal symptoms were enrolled; 33 cats were histologically diagnosed as having alimentary lymphoma and 38 cats as having IBD. Serum samples of total LDH analysis were measured. RESULTS: Gender (P = 0.016) and age (P = 0.046) were found to be significant factors influencing the differentiation of serum total LDH between cats with alimentary lymphoma and those with IBD. Despite low diagnostic accuracy in the overall population (63%), a cut-off value of serum total LDH ranging from 0.85- to 1.04-times the upper reference limit showed good capability (accuracy >80%) of differentiating these two conditions in neutered males and cats younger than 8 years of age (AUC: 0.805, 0.833; sensitivities: 76.9%, 83.3%; specificities: 80.0%, 76.5%; PPV: 76.9%, 55.6%; NPV: 80.0%, 92.9%; respectively). CONCLUSIONS: Although our study showed that gender and age are significant factors in differentiating serum total LDH between cats with alimentary lymphoma and those with IBD, this test had poor diagnostic accuracy in differentiating between these two conditions in the overall population.

Small intestinal ganglioneuromatosis in a dog.

A 9-year-old female neutered collie-cross dog was presented with a 2-month history of persistent diarrhoea, weight loss and intermittent vomiting. Abdominal ultrasonography revealed one loop of jejunum with a markedly thickened and multifocally hyperechoic wall, without loss of wall layering. Laparotomies were performed for biopsy and resection of affected intestine. Histopathological examination revealed small intestinal ganglioneuromatosis (GN). The dog recovered well from surgery and the diarrhoea resolved. Eleven months later the dog has gained weight and remains asymptomatic. This is the first report of small intestinal GN affecting a mature dog, in which pathology was localized to the mucosal lamina propria and surgical treatment resulted in a successful outcome.

Malignant lymphoma of T-cell origin in a Humboldt penguin (Spheniscus humboldti) and a pink-backed pelican (Pelecanus rufescens).

Multicentric T-cell lymphomas were diagnosed in two birds from separate zoological collections: one in a 27-year-old female Humboldt penguin (Spheniscus humboldti) and the second in an adult pink-backed pelican (Pelecanus rufescens). The main clinical sign in the penguin was dysphagia caused by lymphoma formation in the esophagus. Besides the esophageal lymphoma, neoplastic lymphoid cells were observed in the adrenal glands, liver, kidneys, lung, proventriculus, and gizzard. The pelican was found dead without a clinical history. Neoplastic lymphoid cells were observed in the kidneys, liver, pancreas, spleen, ventriculus, and small intestine. Neoplastic cells of the penguin as well as of the pelican were immunoreactive to CD3 antigen, suggesting the lymphomas were of T-cell origin. In both cases, test results were negative for Marek's disease virus, avian leukosis virus, and reticuloendotheliosis virus. In the pelican, a skin melanoma was diagnosed on the left throat pouch in addition to the multicentric T-cell lymphoma.

Extravasation injury associated with parenteral nutrition in a cat with presumptive gastrinomas.

OBJECTIVE: To describe the pathologic consequences of parenteral nutrition (PN) extravasation into the mediastinum of a cat. CASE SUMMARY: An 8-year-old domestic short hair cat with persistent vomiting and anorexia was initiated on PN for nutritional support. PN was being administered at a rate of 12.9 mL/h when inadvertent jugular catheter migration resulted in thrombophlebitis and cellulitis and 40-80 mL of PN extravasated into the SC and mediastinal tissues. The cat was euthanized 36 hours after the extravasation of PN due to poor prognosis related to the gastric complications associated with a presumed primary gastrinoma. Grossly there was excessive mediastinal lymphatic drainage and pronounced edema in the cervical SC and mediastinal tissue. Histopathologic examination of the PN-extravasated area revealed a severe mixed inflammatory reaction, represented by a severe neutrophilic and mild histiocytic infiltrate with lymphoplasmacytic perivascular cuffing. No bacterial agents were observed or cultured from this area. UNIQUE INFORMATION PROVIDED: This is the first case report of a foreign body-type reaction due to extravasation of PN (extravasation injury) in a cat. Extravasation of PN is not without pathologic consequence, and can result in a severe inflammatory reaction in affected tissues.

Adenomatous polyposis coli-deficient zebrafish are susceptible to digestive tract neoplasia.

Truncation of the tumour suppressor adenomatous polyposis coli (APC) constitutively activates the Wnt/beta-catenin signalling pathway. This event constitutes the primary transforming event in sporadic colorectal cancer in humans. Moreover, humans or mice carrying germline truncating mutations in APC develop large numbers of intestinal adenomas. Here, we report that zebrafish that are heterozygous for a truncating APC mutation spontaneously develop intestinal, hepatic and pancreatic neoplasias that are highly proliferative, accumulate beta-catenin and express Wnt target genes. Treatment with the chemical carcinogen 7,12-dimethylbenz[a]anthracene accelerates the induction of these lesions. These observations establish apc-mutant zebrafish as a bona fide model for the study of digestive tract cancer.

Immunohistochemical diagnosis of alimentary lymphomas and severe intestinal inflammation in cats.

Intestinal tissue samples were examined from 32 cats in which a histopathological diagnosis of alimentary lymphoma or multicentric lymphoma affecting the gastrointestinal tract had been made. These samples were re-evaluated histopathologically and serial sections were examined immunohistochemically with antisera specific for the lymphoid markers CD3, CD79a and BLA-36 and for class II molecules of the major histocompatability complex. The cats ranged in age from 4-16 years (median 10.5 years). The main presenting clinical signs were vomiting, diarrhoea and weight loss. The majority of alimentary lymphomas were of the B-cell type (n=15), whereas cases of T-cell lymphoma were fewer in number (n=8). Four cats had lymphoma of a mixed T-and B-cell phenotype. In five of the cats, immunohistochemistry suggested an inflammatory process, in contradiction to the original histopathological diagnosis of lymphoma. Immunolabelling would appear to be a useful adjunct to histopathology in classifying cases of feline alimentary lymphoma, and may help in distinguishing lymphoma from severe intestinal inflammation.

Relation between phenotype of tumor cells and clinicopathology in bovine leukosis.

Thirty-three cases of enzootic bovine leukosis (EBL) and 14 cases of sporadic bovine leukosis (SBL) were examined by immunohistochemistry using 6 monoclonal antibodies against leukocyte differentiation molecules of bovine leukocytes. There were 17 cases of B-1a cell type, 10 cases of B-1b cell type and 6 cases of B-2 cell type in EBL, and 5 cases originating from B cells (B-2 cell type) and 9 cases originating from immature T cells in SBL. The average age for the EBL cases of B-1a cell type was 8.6 years, B-1b cell type was 6.5 years, and of B-2 cell type was 4.5 years. In cases of SBL, immature T cell type patients were younger than B-2 cell type ones. The lymphoma originating from B cells differed from that originating from T cells in morphology. In T cell tumors, the nucleus of tumor cells was round, the edge of the cytoplasm obvious, and tumor cells were sporadically present and proliferated. When compared with T cells, the region among B cells was obscure. But, there was no relation between phenotype and the histologic classification of tumor cells. In EBL, beyond the lymph node, tumors of B-1a and B-1b types had developed in the heart and abomasum, and those of the B-2 type tended to occur in liver. In SBL, B-2 type and T type cells formed tumors in the liver, kidney, thymus, and one case of T-cell type tumor formed on the skin. We would like to propose a new classification of bovine leukosis as EBL, calf type B-cell lymphoma, juvenile T-cell lymphoma and skin type T-cell lymphoma.

Prognostic value of histopathological grading in canine extramedullary plasmacytomas.

The canine extramedullary plasmacytoma (cEMP) has recently been the subject of numerous investigations, indicating that the histomorphologic diagnosis is often difficult because of the variety of morphologic features. Therefore, the objective of this study was to establish a subclassification scheme for cEMPs and to evaluate correlations between the types and malignancy. Retrospectively, 117 cEMPs, all immunohistochemically characterized by a monoclonal immunoglobulin light-chain expression, were collected and assigned to morphologic types. These types were compared using data from a follow-up study on metastasis and tumor recurrence, then compared by proliferation rate, determined by immunohistochemical detection of the antigen Ki-67. Histopathologic typing revealed five different types of cEMPs, ranging from the mature type with typical plasma cells to the polymorphous-blastic type. Between these two forms, three additional types were established: hyaline, cleaved, and asynchronous. Most of the cEMPs were of the cleaved and asynchronous types. In all cEMPs, mononuclear and multinuclear giant cells were present to varying degrees. Although the results of cell proliferation and the follow-up study indicated less benign behavior by the polymorphous-blastic type, the proliferation rate revealed no statistically significant differences among the cEMP types. The clinical data therefore confirmed previous findings that the risk of tumor recurrence and metastasis in general can be classed as low. The established cEMP typing system is probably a very helpful diagnostic tool, although the types cannot be used for a tumor grading system.

Results of chemotherapy for cats with alimentary malignant lymphoma: 21 cases (1993-1997)

OBJECTIVE: To determine clinical and pathologic findings in cats with alimentary malignant lymphoma and results of treatment with a combination of prednisone, L-asparaginase, vincristine, cyclophosphamide, doxorubicin, and methotrexate. DESIGN: Retrospective study. ANIMALS: 21 cats with alimentary malignant lymphoma. PROCEDURE: Medical records were reviewed, and information on signalment, clinical history and signs, previous treatments, and results of laboratory tests, thoracic radiography, and abdominal ultrasonography were obtained. RESULTS: Test results in all cats were negative for FeLV; 3 of 19 were positive for feline immunodeficiency virus. Thirteen tumors were stage III, 7 were stage IV, and 1 was stage V. Diagnosis was confirmed on the basis of microscopic examination of histologic (n = 13) or cytologic (8) specimens. Immunophenotyping was performed on 13 tumors; 10 were T-cell and 3 were B-cell lymphomas. Overall median duration of first remission was 20 weeks. Overall median survival time was 40 weeks. The only factor significantly associated with duration of first remission was whether cats had a complete response following induction chemotherapy; duration of first remission was significantly associated with survival time. Cats tolerated treatment well; only 1 cat had a delay in the treatment schedule because of neutropenia. CLINICAL IMPLICATIONS: Use of a multidrug chemotherapeutic protocol that includes L-asparaginase and doxorubicin results in minimal adverse effects and prolonged survival times in cats with alimentary malignant lymphoma.

Retrospective study of 60 cases of feline lymphosarcoma.

OBJECTIVE: To characterise epidemiological and clinical findings, and diagnostic procedures undertaken, in cats with lymphosarcoma at a veterinary teaching hospital. DESIGN: Retrospective case study. PROCEDURE: Hospital records were reviewed for 7159 cats, sick or healthy, examined during a 10-year period (1984 to 1994). Sixty cats with lymphosarcoma were identified and classified by anatomical location of the tumor. Data on breed, age, sex, clinical signs and diagnostic procedures were collated. RESULTS: The prevalence of feline lymphosarcoma in the hospital population was 0.84%. Siamese cats appeared predisposed to lymphosarcoma but other purebreds were not. Males were somewhat overrepresented amongst affected cats. Similar numbers of cases (12 to 18) were seen in each of the four anatomic categories (multicentric, mediastinal, alimentary and extranodal). Cats with mediastinal lymphosarcoma were mostly young and Siamese. Clinical signs in affected cats were varied, usually multiple and often nonspecific. Two of 22 cases tested positive for feline leukaemia virus antigen in blood and 6 of 13 were positive for feline immunodeficiency virus antibody. CONCLUSIONS: Extranodal lymphosarcoma seemed more prevalent in this study than reported elsewhere. Siamese cats in the study population may have had a genetic predisposition to lymphosarcoma. Limited evidence suggested feline leukaemia virus may be less important, and feline immunodeficiency virus more important, in the local population than indicated in overseas reports. Additional studies are needed to investigate breed predisposition and feline leukaemia virus and feline immunodeficiency virus status in Australian cats with lymphosarcoma.

Experimental reproduction of the papilloma-carcinoma complex of the alimentary canal in cattle.

Bovine papillomavirus type 4 (BPV-4) is the aetiological agent of epithelial papillomas of the upper alimentary canal in cattle. These benign tumours can become a focus for transformation to squamous cell carcinomas in animals feeding on bracken fern. Strong epidemiological evidence suggests that the progression to malignancy is due to the interplay between BPV-4 and mutagenic and immunosuppressing chemicals present in the fern. The carcinomas of the upper alimentary canal are often accompanied by adenomas and adenocarcinomas of the lower intestine and bracken-grazing animals are also heavily immunosuppressed. To elucidate the individual roles and the concerted action of the viral and chemical factors involved in tumorigenesis and malignant conversion, we attempted to reproduce experimentally the cancer syndrome observed in the field. Florid persistent papillomatosis of the upper alimentary canal was reproduced in animals infected with BPV-4 and immunosuppressed either by a diet of bracken or by treatment with azathioprine; cancer of the upper alimentary tract or of the lower intestine developed only in animals infected with virus and fed on bracken fern. As in field cases, BPV-4 DNA was detected in papillomas but not in cancers. We conclude that immunosuppression is necessary for persistence and spread of viral papillomas, that the fern mutagens are responsible for neoplastic conversion of papilloma cells, and that continuous expression of viral functions is not required for the maintenance of the malignant state.

Establishment and characterization of a feline large granular lymphoma cell line expressing interleukin 2 receptor alpha-chain.

A feline large granular lymphocyte (LGL) cell line was established from a cat with an alimentary-form lymphoma. This cell line, designated as FGL, had many large azurophilic granules in the cytoplasm, which were typical to LGL cells. Proviral genome of feline leukemia virus was detected in the chromosomal DNA of FGL cells, and reverse transcriptase activity was also demonstrated in the culture supernatant. Furthermore, we found expression of interleukin 2 (IL-2) receptor alpha-chain on the cell surface of FGL and its natural killer activity against human erythroblastic leukemia cell line, K562.

Spontaneous neoplasms in aged Sprague-Dawley rats.

Incidence of neoplastic lesions in untreated Sprague-Dawley rats (1340 males and 1329 females) used as controls in 17 carcinogenicity studies are tabulated and evaluated. In male rats, the most common neoplasms were benign pheochromocytomas and keratoacanthomas (4.0% in each case) followed by pancreatic islet cell adenomas (3.7%), thyroid parafollicular cell adenomas (3.6%), fibromas and squamous cell papillomas of the skin and hepatocellular adenomas (2.0% in each), malignant lymphoma lymphocytic (1.9%), histiocytic sarcomas (1.4%), and adrenal cortical adenomas (1.2%). In female rats, the most common neoplasms were of mammary gland origin (31.3%: fibroadenoma 19.0%, adenocarcinomas 8.8%, and adenomas 3.5%) followed by thyroid parafollicular cell adenomas (2.9%), uterine endometrial stromal polyps (2.6%), adrenal cortical adenomas (1.9%), malignant lymphoma lymphocytic (1.6%), fibromas in the skin (1.3%), and pancreatic islet cell adenoma (1.1%). Metastases were observed from pheochromocytomas, hepatocellular carcinomas, nephroblastomas, renal pelvis transitional cell carcinoma, interstitial cell tumor and seminoma of the testes, Zymbal's gland adenocarcinomas, and mammary adenocarcinomas.

Bracken as a risk factor in digestive tract tumors: state of the art.

The present knowledge of the risks of exposure to bracken fern (Pteridium aquilinum) is presented. The history of research on bracken toxicity is reviewed, covering the animal observations and chemical analysis, and focusing on the carcinogenic effects in various animal species and on the nature and biological properties of the known carcinogens. The implications of human exposure to bracken are inferred, the possible ways of contamination are discussed, and the existing epidemiological studies are commented. Bracken contribution to digestive tract tumors is deduced, and the need of further research on the plant toxicity and detailed epidemiological investigations with the exposed population is stressed.