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1.
J Small Anim Pract ; 53(12): 709-13, 2012 Dec.
Article in English | MEDLINE | ID: mdl-23020172

ABSTRACT

An 18-month-old male basset hound was presented with vomiting, diarrhoea and depression. Abdominal ultrasonography revealed a mass in the left kidney. An ultrasound-guided core-biopsy indicated aggregates of spindle cells, but did not allow a definitive diagnosis. Nephrectomy was performed after a period of six months, when ultrasound examination revealed a slight increase in mass dimensions. Histologically the mass was composed of neoplastic spindle cells forming interlacing fascicles, bundles and whorls, within a loose myxoid to dense collagenous stroma. Immunohistochemically neoplastic cells were positive for vimentin and smooth muscle actin. Based on these findings the tumour was diagnosed as a congenital mesoblastic nephroma, classical variant. After a two-and-a-half-year follow-up the dog was clinically healthy, indicating a benign behaviour. To the authors' knowledge, this report describes the first case of canine congenital mesoblastic nephroma successfully treated surgically, with a reasonable postsurgical follow-up.


Subject(s)
Dog Diseases/congenital , Kidney Neoplasms/veterinary , Nephroma, Mesoblastic/veterinary , Animals , Dog Diseases/diagnosis , Dog Diseases/surgery , Dogs , Immunohistochemistry/veterinary , Kidney Neoplasms/congenital , Kidney Neoplasms/diagnosis , Kidney Neoplasms/surgery , Male , Nephroma, Mesoblastic/diagnosis , Nephroma, Mesoblastic/surgery
2.
Cell Prolif ; 39(6): 611-22, 2006 Dec.
Article in English | MEDLINE | ID: mdl-17109643

ABSTRACT

A new murine cell line, named GFPneu, was established from a mammary adenocarcinoma arising in double transgenic MMTVneu x CMV-GFP mice. Breast tumours develop in 100% of females after 2 months latency, as a result of the over-expression of the activated rat neu oncogene in the mammary glands. All tissues, and in particular the breast tumours, express the GFP protein. This cell line was tumorigenic when inoculated into nude mice and the derived tumours showed the same histological features as the primaries from which they were isolated. Their histopathology reproduces many characteristics of human breast adenocarcinomas, in particular their ability to metastasize. The GFP marker allows us to visualize the presence of lung metastases in fresh tissues immediately, to confirm the histopathology. From a lung metastatic fluorescent nodule, we derived a further cell line, named MTP-GFP, which we also characterized. These two cell lines could be useful to study the role played by the neu oncogene in the maintenance of the transformed phenotype, in the metastatic process, to test novel therapeutic strategies to inhibit primary tumour growth and to observe the generation of distant metastases.


Subject(s)
Adenocarcinoma/genetics , Cell Line, Tumor , Genes, erbB-2/genetics , Green Fluorescent Proteins/genetics , Mammary Neoplasms, Animal/genetics , Adenocarcinoma/secondary , Animals , Female , Gene Expression Regulation, Neoplastic , Lung Neoplasms/genetics , Lung Neoplasms/secondary , Mammary Neoplasms, Animal/pathology , Mice , Mice, Nude , Mice, Transgenic , Telomere
3.
J Vet Intern Med ; 20(4): 822-7, 2006.
Article in English | MEDLINE | ID: mdl-16955803

ABSTRACT

Infection with Helicobacter spp. is increasingly linked with hepatobiliary inflammation and neoplasia in people and in a variety of animals. We sought to determine if Helicobacter species infection is associated with cholangiohepatitis in cats. Deoxyribonucleic acid was extracted from tissue blocks from cats with cholangiohepatitis (32), noninflammatory liver disease (13), and cats with normal liver histology (4). Deoxyribonucleic acid was polymerase chain reaction-amplified with 2 sets of Helicobacter genus-specific primers, gel purified, and sequenced. Polymerase chain reaction-positive hepatic tissue was further examined with Steiner's stain, immunocytochemistry for Helicobacter species, and eubacterial fluorescent in situ hybridization. Gastric tissues of cats with known Helicobacter infection status served as controls for deoxyribonucleic acid extraction and sequence comparison. Helicobacter species were detected in 2/32 cats with cholangiohepatitis, and 1/17 controls. Sequences had 100% identity with Helicobacter species liver, Helicobacter pylori, and Helicobacter fenelliae/cinaedii in a cat with suppurative cholangitis, Helicobacter species liver, Helicobacter pylori, and Helicobacter nemistrineae in a cat with mild lymphocytic portal hepatitis, and Helicobacter bilis in a cat with portosystemic vascular anomaly. In contrast, sequences from gastric biopsies showed highest homology (99-100%) to "Helicobacter heilmannii," Helicobacter bizzozeronii, Helicobacter felis, and Helicobacter salomonis. Fluorescent in situ hybridization revealed a semicurved bacterium, with Helicobacter-like morphology, in an intrahepatic bile duct of the cat with suppurative cholangitis. This study has identified Helicobacter deoxyribonucleic acid in 2/32 cats with cholangiohepatitis and 1/13 cats with noninflammatory liver disease. Deoxyribonucleic acid sequences of hepatic Helicobacter species were distinct from those found in the stomach and are broadly consistent with those identified in cat intestine and bile, and hepatobiliary disease in people and rodents.


Subject(s)
Biliary Tract Diseases/veterinary , Cat Diseases/microbiology , Helicobacter/physiology , Animals , Biliary Tract Diseases/microbiology , Cats , Female , Helicobacter/isolation & purification , Male
4.
J Vet Med A Physiol Pathol Clin Med ; 51(6): 280-3, 2004 Aug.
Article in English | MEDLINE | ID: mdl-15485563

ABSTRACT

Neosporosis, caused by the protozoan parasite Neospora caninum, is a serious cause of bovine abortion, stillbirth and perinatal death. This paper reports a clinical neosporosis in a 3-week-old fallow deer (Dama dama). The fawn was full term and appeared normal at birth. Histological lesions consisted of a multifocal necrotizing and granulomatous meningoencephalomyelitis, with intralesional protozoal cysts. Positive immunohistochemical staining and characteristic PCR products confirmed the diagnosis of N. caninum infection.


Subject(s)
Coccidiosis/veterinary , Deer , Meningoencephalitis/veterinary , Neospora/isolation & purification , Animals , Animals, Newborn , Animals, Wild , Coccidiosis/diagnosis , Coccidiosis/pathology , DNA Primers , DNA, Protozoan/analysis , Diagnosis, Differential , Female , Immunohistochemistry/veterinary , Meningoencephalitis/diagnosis , Meningoencephalitis/pathology , Neospora/genetics , Polymerase Chain Reaction/veterinary
5.
Gene Ther ; 10(22): 1903-9, 2003 Oct.
Article in English | MEDLINE | ID: mdl-14502219

ABSTRACT

Treatments available to women with locally advanced breast cancer are unsatisfactory, since most patients succumb to metastatic spread. Therefore, there is a need to devise novel therapeutic combinations that effectively inhibit metastatization and to test them in animal models of breast cancer showing strong similarities with their human counterpart, including the ability to give rise to metastases. With these considerations in mind, tamoxifen (TAM), 4-hydrotamoxifen (4-HT) or liposome-complexed DNA constructs coding for antiangiogenic/anti-invasion proteins (angiostatin, TIMP-2, IFN-alpha(1), sFLT-1) were individually administered to MMTVneu transgenic mice. Significant inhibition of primary tumor growth was obtained with TAM (40% inhibition, P=0.049), angiostatin (85% inhibition, P=0.001) and TIMP-2 (60% inhibition, P=0.015). No lung metastasis was observed in any of these treated mice at 5 months, compared with a rate of 70% in control groups. These observations were the basis for designing a combined treatment with all these compounds. The association of angiostatin, TIMP-2 and TAM was greatly effective at the primary tumor level (90% inhibition, P=0.01). Moreover, all the mice treated with this association were metastasis free at a time point (6 months) in which seven out of nine control mice were either dead from disseminated cancer or showed lung metastasis. This combined therapy could become an important component of anticancer therapy in humans.


Subject(s)
Breast Neoplasms/therapy , Estrogen Receptor Modulators/therapeutic use , Genetic Therapy/methods , Lung Neoplasms/prevention & control , Lung Neoplasms/secondary , Tamoxifen/therapeutic use , Angiogenesis Inhibitors/genetics , Angiostatins , Animals , Combined Modality Therapy , Extracellular Matrix Proteins/genetics , Female , Gene Expression , Genes, erbB-2 , Genetic Vectors/administration & dosage , Interferon-alpha/genetics , Mice , Mice, Transgenic , Models, Animal , Myosin Heavy Chains , Nonmuscle Myosin Type IIB , Peptide Fragments/genetics , Plasminogen/genetics , Tissue Inhibitor of Metalloproteinase-2/genetics
6.
Gene Ther ; 9(19): 1338-41, 2002 Oct.
Article in English | MEDLINE | ID: mdl-12224018

ABSTRACT

Breast tumor growth and metastasization are both hormone-sensitive and angiogenesis-dependent. Recent work carried out in our laboratory on a transgenic model of breast cancer displaying many similarities to its human counterpart, has shown that liposome-mediated angiostatin cDNA delivery partially inhibits both local and metastatic growth. However, it is now recognized that anti-angiogenesis strategy alone cannot completely arrest tumor growth and spread, and this led to the suggestion that approaches based on different molecular mechanisms could usefully be combined. In the present work, we investigated whether tamoxifen, a classical antiestrogen agent widely used in human therapy, could improve the results obtained with angiostatin alone. Further reduction of local growth was achieved with the combined regimen with respect to angiostatin or tamoxifen alone, while, as expected, no metastatic growth was detected in either group. We therefore conclude that a combination of angiogenesis inhibitors with antiestrogen drugs might be useful in humans and that other associations between conventional and gene transfer-mediated therapy are worth investigating and will soon become important components of anticancer therapy.


Subject(s)
Genetic Therapy/methods , Mammary Neoplasms, Experimental/therapy , Neovascularization, Pathologic/prevention & control , Peptide Fragments/genetics , Plasminogen/genetics , Tamoxifen/therapeutic use , Angiogenesis Inhibitors/genetics , Angiogenesis Inhibitors/metabolism , Angiostatins , Animals , Antineoplastic Agents, Hormonal/therapeutic use , Combined Modality Therapy , DNA, Complementary/genetics , Female , Lung Neoplasms/prevention & control , Lung Neoplasms/secondary , Mammary Neoplasms, Experimental/blood supply , Mammary Neoplasms, Experimental/drug therapy , Mice , Mice, Transgenic , Peptide Fragments/metabolism , Plasminogen/metabolism
7.
Cancer Gene Ther ; 8(7): 491-6, 2001 Jul.
Article in English | MEDLINE | ID: mdl-11498770

ABSTRACT

The effect of local and systemic delivery of the angiostatin gene on human melanoma growth was studied in nude mice. Liposome-coated plasmids carrying the cDNA coding for murine and human angiostatin (CMVang and BSHang) were injected weekly, locally or systemically, in mice transplanted with melanoma cells. The treatment reduced melanoma growth by 50% to 90% compared to that occurring in control animals treated with liposome-coated plasmid carrying the lacZ gene or in untreated controls. The growth of both locally injected and controlateral uninjected tumors in mice bearing two melanoma grafts was significantly suppressed after intratumoral treatment. Tumor growth inhibition was also observed in mice treated by intraperitoneal delivery, suggesting that angiostatin gene therapy acts through a systemic effect. Both melanoma growth suppression and delay in the onset of tumor growth were observed in treated mice. PCR performed on tumors and normal tissues showed that the lipofected DNA was present in tissues from treated mice, and angiostatin expression was demonstrated by RT-PCR. Histopathological analysis of melanoma nodules revealed an increase in apoptotic cells and a reduction in vessel density in tumors from treated mice. Our results suggest that systemic, liposome-mediated administration of genes coding for antiangiogenic factors represents a promising strategy for melanoma treatment in humans.


Subject(s)
Genetic Therapy/methods , Melanoma/therapy , Peptide Fragments/genetics , Plasminogen/genetics , Angiostatins , Animals , Cell Division , DNA/metabolism , DNA, Complementary/metabolism , Humans , Lac Operon/genetics , Liposomes/metabolism , Mice , Mice, Nude , Neoplasm Transplantation , Neovascularization, Pathologic , Peptide Fragments/biosynthesis , Plasminogen/biosynthesis , Reverse Transcriptase Polymerase Chain Reaction , Time Factors , Tumor Cells, Cultured
8.
Gene Ther ; 8(1): 67-70, 2001 Jan.
Article in English | MEDLINE | ID: mdl-11402303

ABSTRACT

Tumor growth and metastasis are angiogenesis-dependent. The possibility of inhibiting tumor growth by interfering with the formation of new vessels has recently raised considerable interest. We previously reported that it is possible to inhibit primary tumor growth and metastasis in a transgenic model of spontaneous breast tumor, which shows many similarities to its human counterpart (including ability to metastasize) by intratumoral administration of a DNA construct carrying the murine angiostatin cDNA driven by liposomes. Here we report that it is also possible to achieve this goal by a systemic (intraperitoneal) delivery of therapeutic DNA constructs carrying genes coding for mouse and human anti-angiogenic factors which include angiostatin, endostatin and TIMP-2. These findings may be relevant to the design of therapeutic interventions in humans.


Subject(s)
Adenocarcinoma/therapy , Angiogenesis Inhibitors/genetics , Genetic Therapy/methods , Mammary Neoplasms, Experimental/therapy , Neovascularization, Pathologic/prevention & control , Adenocarcinoma/blood supply , Adenocarcinoma/pathology , Adenocarcinoma/secondary , Animals , Female , Humans , Liposomes , Lung Neoplasms/prevention & control , Lung Neoplasms/secondary , Mammary Neoplasms, Experimental/blood supply , Mammary Neoplasms, Experimental/pathology , Mice , Mice, Transgenic
9.
J Vet Diagn Invest ; 13(1): 3-12, 2001 Jan.
Article in English | MEDLINE | ID: mdl-11243359

ABSTRACT

Detailed histopathological evaluation of the gastric mucosa of Helicobacter-infected cats is complicated by the difficulty of recognizing Helicobacter organisms on hematoxylin and eosin (HE)-stained sections and the ability of multiple Helicobacter species to infect cats. In this study, the presence and localization of different species of Helicobacter in the stomachs of cats was investigated using silver staining and immunohistochemistry. Five groups containing 5 cats each were established (group 1: urease negative and Helicobacter free; groups 2, 3, 4, and 5: urease positive and infected with Helicobacter heilmannii, unclassified Helicobacter spp., Helicobacter felis, and Helicobacter pylori, respectively). Gastric samples were evaluated by HE and silver staining and by immunohistochemistry with 3 different anti-Helicobacter primary antibodies. Helicobacter were detected by Steiner stain in all infected cats at the mucosal surface, in the lumen of gastric glands, and in the cytoplasm of parietal cells. In silver-stained sections, H. pylori was easily differentiated from H. felis, H. heilmannii, and unclassified Helicobacter spp., which were larger and more tightly coiled. No organisms were seen in uninfected cats. Helicobacter antigen paralleled the distribution of organisms observed in Steiner-stained sections for 2 of the 3 primary antibodies tested. The antisera were not able to discriminate between the different Helicobacter species examined. A small amount of Helicobacter antigen was present in the lamina propria of 3 H. pylori-, 3 H. felis-, and 1 H. heilmannii-infected cat. Minimal mononuclear inflammation was present in uninfected cats and in those infected with unclassified Helicobacter spp. and H. heilmannii cats. In H. felis-infected cats, lymphoid follicular hyperplasia with mild pangastric mononuclear inflammation and eosinophilic infiltrates were present. The H. pylori-infected cats had severe lymphoid follicular hyperplasia and mild to moderate mononuclear inflammation accompanied by the presence of neutrophils and eosinophils. These findings indicate that Steiner staining and immunohistochemistry are useful for detecting Helicobacter infections, particularly when different Helicobacter species can be present. Monoclonal antibodies specific for the different Helicobacter species could be important diagnostic aids. There appear to be differences in the severity of gastritis in cats infected with different Helicobacter species.


Subject(s)
Cat Diseases/microbiology , Gastritis/veterinary , Helicobacter Infections/veterinary , Helicobacter/isolation & purification , Animals , Antibodies, Monoclonal , Cat Diseases/genetics , Cat Diseases/pathology , Cats , DNA, Bacterial/analysis , Diagnosis, Differential , Female , Gastritis/microbiology , Gastritis/pathology , Helicobacter/immunology , Helicobacter Infections/immunology , Helicobacter Infections/pathology , Immunohistochemistry , Intestinal Mucosa/pathology , Male , Polymerase Chain Reaction
10.
J Biomater Sci Polym Ed ; 10(2): 247-57, 1999.
Article in English | MEDLINE | ID: mdl-10091934

ABSTRACT

In this study, thirty-nine patients were examined. All of them suffered from hip joint prostheses loosening and underwent revision surgery. Bioptic samples were collected at the interface between bone and implant either at the stem or cotyle level. Immunohistochemistry was performed to detect IL-1alpha, IL-1beta, IL-6 and TNF, cytokines that directly cause bone resorption and indirectly induce synthesis of other bone resorbing cytokines. Quantitative analysis of the positive cells and correlation with clinical data was performed. It resulted that there is a great variability in positive cells for cytokines according to the harvest site; anyway, cytokines tend to be higher in patients carrying a joint prosthesis with polyethylene acetabular component and it is associated with plastic wear particles, even though there is no direct correlation between wear amount and cytokine levels. There is a statistically significant negative correlation between metal wear and a cytokine (IL-6); cytokines levels do not depend on the implant time to failure and do not correlate with pain score. As expected, cytokines levels tend to be lower in subjects being treated with non-steroidal antiinflammatory drugs. It can be concluded that plastic wear is the factor inducing the highest cytokine levels in the tissues around the prosthesis at the interface; cytokines that are an indicator of osteolysis risk.


Subject(s)
Arthroplasty, Replacement, Hip/adverse effects , Cytokines/biosynthesis , Adult , Aged , Aged, 80 and over , Bone Cements/metabolism , Female , Hip Joint/cytology , Hip Joint/drug effects , Humans , Immunohistochemistry , Interleukin-1/biosynthesis , Interleukin-1/metabolism , Interleukin-6/biosynthesis , Interleukin-6/metabolism , Male , Middle Aged , Tissue Distribution , Tumor Necrosis Factor-alpha/biosynthesis , Tumor Necrosis Factor-alpha/immunology , Tumor Necrosis Factor-alpha/metabolism
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