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Biomedical subjects

W F Robinson

Publications and source records attributed to W F Robinson.

At least 19 recordsLinked to original sources
PubMed2004

Characterisation of three novel canine osteosarcoma cell lines producing high levels of matrix metalloproteinases.

Three canine osteosarcoma cell lines were established from spontaneous pelvic and radial osteosarcomas. The cell populations cultured exhibited characteristics of malignancy and consisted of adherent, pleomorphic, mostly large spindle-shaped or polyhedral cells, characterised by the presence of numerous cytoplasmic granules and vacuoles. The main ultrastructural features included the presence of abundant rough endoplasmic reticulum and numerous cytoplasmic vesicles, deposit vacuoles and small cytoplasmic protrusions. Zymography showed that the cell lines produce high levels of MMP-2 and MMP-9, enzymes directly involved in crucial aspects of the metastatic process. Consistent with their osteoblastic lineage and malignant phenotype, all cell lines were immunoreactive to vimentin, osteopontin, PCNA, p53, MMP-2 and MMP-9, while they were negative for cytokeratin, desmin, SMA, Factor VIII, NSE, GFAP, Rb and p21 protein. No retroviral particles or RNA were detected ultrastructurally or with RT-PCR, although the possibility of viral involvement in osteosarcoma cannot be excluded. The new cell lines provide excellent in vitro models that may allow further studies on the pathobiology of canine osteosarcoma to be undertaken.

Alkaline Phosphatase↗
PubMed2003

Clinical and pathologic relevance of p53 index in canine osseous tumors.

The clinicopathologic value of the immunohistochemical (IHC) expression of p53 protein was evaluated in 167 canine osseous tumors. p53 staining frequency and intensity in tumor cells was expressed as a p53 index. p53 index was significantly higher in osteosarcomas than in other sarcomas, chondrosarcoma, multilobular tumor of bone, and tumors initially misdiagnosed as osteosarcomas as well as in appendicular versus axial and in distal versus proximal osteosarcomas. A strong correlation is demonstrated between the p53 index and a range of clinicopathologic parameters in osteosarcoma, including the tumor site, histologic grade and score, mitotic index, degree of tumor necrosis, and pleomorphism. Chondroblastic osteosarcomas had significantly higher and telangiectatic osteosarcomas significantly lower p53 index than did osteosarcomas belonging to other histopathologic subtypes, a fact that tends to reinforce the perception of these osteosarcomas as distinct clinicopathologic entities. Entire males had higher p53 index than did neutered males. p53 index was higher in Rottweilers than in Great Danes and Terriers, confirming breed susceptibilities to osteosarcoma. p53 index showed no association with age, primary or secondary site status, or the presence of metastases or other tumor types. Biopsy samples had a higher p53 index than did postmortem samples, either because of differences in sample processing or the possibility that p53 overexpression is more evident at the earlier stages of osteosarcoma pathogenesis, presumably represented by the biopsy material. IHC examination for p53 and the derived index has the potential to be used as an additional diagnostic tool and prognostic indicator for osseous tumors.

Animals↗
PubMed2003

Matrix metalloproteinase-2 and -9 involvement in canine tumors.

Matrix metalloproteinases (MMPs) are a family of enzymes implicated in the degradation and remodeling of extracellular matrix and in vascularization. They are also involved in pathologic processes such as tumor invasion and metastasis in experimental cancer models and in human malignancies. We used gelatin zymography and immunohistochemistry to determine whether MMP-2 and MMP-9 are present in canine tumors and normal tissues and whether MMP production correlates with clinicopathologic parameters of prognostic importance. High levels of pro-MMP-9, pro-MMP-2, and active MMP-2 were detected in most canine tumors. Significantly higher MMP levels were measured in canine tumors than in nontumors, malignancies had higher MMP levels than benign tumors, and sarcomas had higher active MMP-2 than carcinomas. Cartilaginous tumors produced higher MMP levels than did nonsarcomatous malignancies, benign tumors, and normal tissues, and significantly greater MMP-2 than osteosarcomas and fibrosarcomas. Pro-MMP-9 production correlated with the histologic grade of osteosarcomas. The 62-kd form of active MMP-2 was detected only in high-grade, p53-positive, metastatic malignancies. Zymography proved to be a sensitive and quantitative technique for the assessment of MMP presence but has the limitation of requiring fresh tissue; immunohistochemistry is qualitative and comparatively insensitive but could be of value in archival studies. MMP presence was shown in a range of canine tumors, and their link to tumor type and grade was demonstrated for the first time. This study will allow a substantially improved evaluation of veterinary cancer patients and provides baseline information necessary for the design of clinical trials targeting MMPs.

Animals↗
PubMed2002

Productive infection of the bone marrow cells in feline immunodeficiency virus infected cats.

Bone marrow stromal cells (BMSC) from cats experimentally infected with feline immunodeficiency virus (FIV) were shown to contain FIV provirus using polymerase chain reaction and viral products were detected in culture supernatant using reverse transcriptase and enzyme linked immunosorbent assay techniques. Peripheral blood mononuclear cells from FIV-free cats co-cultured with infected bone marrow cells became productively infected with FIV. Such evidence supports the hypothesis that BMSC are a reservoir for FIV. Furthermore, BMSC produced virions capable of infecting susceptible cells and may represent an important source of infectious virus to cells of the macrophage lineage and/or hemopoietic progenitor cells, both of which ultimately become widely disseminated throughout the body.

Animals↗
PubMed2001

Characterisation of lymphosarcomas in Australian cats using polymerase chain reaction and immunohistochemical examination.

OBJECTIVE: To examine tumour tissue of cats with lymphosarcoma for the presence of feline leukaemia virus and feline immunodeficiency virus and analyse the immunophenotype of the tumours. DESIGN: A retrospective study of feline lymphosarcoma cases. METHODS: Formalin-fixed, paraffin-embedded tumour tissue of 14 feline lymphosarcomas was examined for the presence of feline leukaemia virus and feline immunodeficiency virus by polymerase chain reaction and immunohistochemistry. Using polyclonal and monoclonal antibodies against T and B lymphocytes, the phenotypic expression of the tumours was characterised. RESULTS: No feline leukaemia virus antigen or proviral sequences were detected. Feline immunodeficiency virus proviral sequences were detected in two cases by polymerase chain reaction. Immunophenotyping of all 14 cases resulted in seven cases being classified as B-cell phenotype, four as T-cell phenotype, and the remaining three undetermined. CONCLUSIONS: In contrast to previous reports overseas, our results suggest that feline leukaemia virus infection appears to be an infrequent cause of lymphosarcoma in the cats that were necropsied. Feline immunodeficiency virus may have a role in lymphomagenesis. The potential role of feline immunodeficiency virus needs to be explored in more depth. Compared with most previous reports, B-cell tumours were more common than T-cell tumours in this series of cats.

Animals↗
PubMed2000

Systemic mycosis due to Aspergillus deflectus in a dog.

A 4-year-old, entire female, German Shepherd Dog was referred with a 3-month history of right foreleg lameness that partially responded to nonsteroidal anti-inflammatory and antimicrobial therapy. The bitch lost weight, was polydipsic and had reduced exercise tolerance. On referral, the animal was in poor condition, pyrexic and exhibited moderate pain on full extension of the right shoulder. Blood, urine and joint fluid were obtained and radiographs were taken of the right shoulder and chest. The bitch was lymphopaenic, hyperfibrinogenaemic, hyperglobulinaemic, mildly azotaemic, mildly proteinuric and isosthenuric. Branching fungal hyphae were present in the urine. On radiography, the thorax contained a large ventral mediastinal mass and the humeral head had extensive areas of radiolucency. An aspirate from the right humeroscapular joint exhibited branched fungal hyphae and numerous neutrophils and macrophages. A diagnosis of disseminated mycosis was made and euthanasia was performed. At necropsy, numerous caseating granulomas were present, especially in the kidneys, adrenal glands, heart and lymph nodes. Extensive osteomyelitis involved the head of the right humerus, the sternebrae and the fifth intervertebral disc. Fungal hyphae were detected in sections of granulomas in all affected organs and a diagnosis of disseminated fungal granulomatosis was made. Aspergillus deflectus was readily isolated from affected lymph nodes, but confirming its identity as A deflectus using standard procedures proved difficult. The identity of the fungus was finally confirmed by sequencing part of the 185 rRNA of the isolate. This is the first report in Australia of a disseminated mycosis caused by A deflectus. Previously, the involvement of A deflectus as a cause of disseminated mycosis was limited to 5 cases from the West Coast of the USA, four of which occurred in German Shepherd Dogs.

Animals↗
PubMed2000

The nucleotide sequence of koala (Phascolarctos cinereus) retrovirus: a novel type C endogenous virus related to Gibbon ape leukemia virus.

A novel retrovirus, morphologically consistent with mammalian C-type retroviruses, was detected by electron microscopy in mitogen-stimulated peripheral blood mononuclear cell cultures from 163 koalas and in lymphoma tissue from 3 koalas. PCR amplified provirus from the blood and tissues of 17 wild and captive koalas, and reverse transcriptase-PCR demonstrated viral mRNA, viral genomic RNA, and reverse transcriptase activity in koala serum and cell culture supernatants. Comparison of viral sequences derived from genomic DNA and mRNA showed identity indicative of a single retroviral species-here designated koala retrovirus (KoRV). Southern blot analysis of koala tissue genomic DNA using labelled KoRV probes demonstrated banding consistent with an endogenous retrovirus. Complete and apparently truncated proviruses were detected in DNA of both clinically normal koalas and those with hematopoietic disease. KoRV-related viruses were not detected in other marsupials, and phylogenetic analysis showed that KoRV paradoxically clusters with gibbon ape leukemia virus (GALV). The strong similarity between GALV and KoRV suggests that these viruses are closely related and that recent cross-host transmission has occurred. The complete proviral DNA sequence of KoRV is reported.

Animals↗
PubMed1999

Feline peripheral blood mononuclear cells express message for both CXC and CC type chemokine receptors.

The feline immunodeficiency virus (FIV) causes a disease in cats similar in clinical presentation and disease progression to that of HIV and AIDS. It is now known that, for HIV infection, as well as primary binding of virus to the CD4 receptor, entry and infection of cells requires coreceptors which are members of the chemokine group of G-protein coupled receptors. Because of the similarity of HIV and FIV, we hypothesised that coreceptors are required for the entry and infection of cells by FIV. Using a feline cDNA library derived from a feline IL-2 sensitive lymphocyte cell line, we identified the presence of message for both CC and CXC chemokine receptors. The feline CXCR4 has been shown to facilitate fusion by FIV [44] and we suggest that the feline CCR5 receptor mediates infection of feline cells by M-tropic strains of FIV.

Amino Acid Sequence↗
PubMed1999

T-cell-rich B-cell lymphoma in the cat.

The clinical and pathological features of eight cases of feline T-cell-rich B-cell lymphoma are described. The disease occurred in older cats (mean age 11.4 +/- 3.9 years), which on initial examination generally showed enlargement of a single submandibular or cervical lymph node. After excision, there was no recurrence of the lesions at 6 months in three cats. In one further case, however, the lesion had recurred 6 months later; it was again excised but recurred after an additional 6 months. Microscopically, there was effacement of normal lymph node architecture by a nodular (n = 4) or diffuse (n = 4) proliferation of small to blastic lymphocytes, accompanied by a characteristic population of bizarre giant, or multinucleate, cells. The mitotic rate was low and mitoses were restricted to the atypical population. Immunophenotyping revealed the smaller lymphocytes to be a mixture of CD3+ MHC Class II+ T lymphocytes and BLA36+CD79variable MHC Class IIvariable B lymphocytes. The atypical cells were of the B-cell lineage (BLA36+MHC Class IIvariable). Polymerase chain reaction analysis revealed no proviral DNA products of feline leukaemia virus or feline immunodeficiency virus in tissue from any tumour, confirming that these neoplasms were not associated with either virus. The clinical, histological and immunophenotypic findings in these cats were identical with those of "nodular lymphocyte predominance (lymphocytic and histiocytic/L&H) Hodgkin's disease" in man.

Age Factors↗
PubMed1999

Polycystic kidney disease in bull terriers: an autosomal dominant inherited disorder.

The prevalence, mode of inheritance and urinalysis findings in Bull Terriers with polycystic kidney disease were assessed by screening 150 clinically normal dogs. The disorder was diagnosed in 39 dogs on the basis of renal ultrasound results and family history of the disease. In equivocal cases confirmation required gross and histopathological renal examination. Necropsy was performed on nine affected dogs and the kidneys from another five affected animals were also examined. Renal cysts were usually bilateral, occurred in cortex and medulla and varied from less than 1 mm to over 2.5 cm in diameter. Cysts were lined by epithelial cells of nephron origin. Abnormal urine sediment and proteinuria were common in affected dogs. The disease appears to be inherited in a highly penetrant autosomal dominant manner.

Animals↗
PubMed1997

Antibody reactivity to the transmembrane protein of the caprine arthritis encephalitis virus correlates with severity of arthritis: no evidence for the involvement of epitope mimicry.

Serum and synovial antibody reactivities of caprine arthritis encephalitis virus (CAEV) infected goats were assessed by Western blotting against purified CAEV antigen and the greatest intensity of reactivity in the serum of arthritic goats was to the gp45 transmembrane protein (TM). The extracytoplasmic domain of the TM gene was cloned into a pGEX vector and expressed in Escherichia coli as a glutathione S transferase fusion protein (GST-TM). This clone was found to be 90.5 and 89.2% homologous to published sequences of CAEV TM gene. Serum of 16 goats naturally infected with CAEV were examined by Western blotting for reactivity to the fusion protein. Antibody reactivity to the GST-TM correlated with clinically detectable arthritis (R = 0.642, P < or = 0.007). The hypothesis that the immune response to the envelope proteins of the CAEV contributes to the severity of arthritis in goats naturally infected with CAEV via epitope mimicry was tested. Antibodies from 5 CAEV infected goats were affinity purified against the GST-TM fusion protein and tested for cross-reactivity with a series of goat synovial extracts and proteogylcans. No serum antibody response or cross-reactivity of affinity purified antibodies could be detected. Peptides of the CAEV SU that were predicted to be linear epitopes and a similar heat shock protein 83 (HSP) peptide identified by database searching, were synthesized and tested for reactivity in CAEV goats using ELISA, in vitro lymphocyte proliferation and delayed type hypersensitivity (DTH) assays. Peripheral blood lymphocytes from 10 of 17 goats with long term natural CAEV infections proliferated in vitro in response to CAEV and in vivo 3 of 7 CAEV infected goats had a DTH reaction to CAEV antigen. However, none of the peptides elicited significant cell mediated immune responses from CAEV infected goats. No antibody reactivity to the SU peptides or HSP peptide was found. We observed that the antibody reactivity to the CAEV TM protein associated with severity of arthritis however epitope mimicry by the envelope proteins of CAEV is unlikely to be involved.

Animals↗
PubMed1997

Transmission of human and feline immunodeficiency viruses via reused suture material.

Several documented cases of human immunodeficiency virus (HIV) infection have involved unconventional or unknown modes of transmission of the virus. Some such cases have occurred within a surgical setting. We investigated the potential for transmission of HIV on suture material that had been reused following passage through an HIV-infected patient. Initial experiments were conducted in vitro using HIV. To provide stronger evidence that HIV could be transmitted via this route, further experiments were undertaken in vivo using a feline immunodeficiency virus (FIV)/cat model. Both methods indicated the possibility of transmission of virus if suture materials were reused.

Animals↗
PubMed1996

Quasispecies and naturally occurring superinfection in feline immunodeficiency virus infection.

Analysis of individual clones containing the V1 and V2 domains of the segment of the FIV env gene present in a naturally infected cat (T) was carried out. The polymerase chain reaction (PCR) was used to amplify proviral FIV DNA extracted from peripheral blood mononuclear cells (PBMCs) obtained in October 1994 from this cat. The PCR products were cloned and the DNA sequences determined for 11 clones. Sequences obtained were aligned with sequences corresponding to FIV isolates (T90, T91, T92) previously obtained from the same cat in 1990, 1991 and 1992. Phylogenetic analysis was performed which included consensus sequences of another Australian isolate, N91, as well as UK, US, Swiss and Japanese isolates of FIV. All clones varied from each other, and none of these clones was identical to the consensus sequences of the isolates obtained previously from the same cat (the T-series). However, most of these clones appeared to have originated from the ancestor of the most recent isolate (T92). In addition, 2 of the clones (7&11) are closely related to another Australian isolate N91, obtained from a different cat (N) in 1991. Because these two cats (T and N) were housed together for at least 3 years (1990-1993) it is suggested that the first cat (T) has become superinfected with an isolate from a second cat (N) under natural conditions. The identification of clones of differing sequences, which were not identical to each other nor to their ancestors, emphasises the rapid mutation of lentiviruses within the env region, and the difficulty of developing an effective FIV vaccine. More importantly, the possibility of natural superinfection with FIV in cats has implications for the development of a successful lentiviral vaccine.

Amino Acid Sequence↗
PubMed1995

Quantification of lymphadenopathy in experimentally induced feline immunodeficiency virus infection in domestic cats.

Nine cats experimentally infected with feline immunodeficiency virus (FIV) and six FIV-negative cats were necropsied to assess the effect of FIV infection on lymph nodes. The FIV infected cats were inoculated with 10(5) TCID50 21-22 weeks previously. The combined weights of all lymph nodes and the combined lymph node to organ weight ratios were significantly greater in FIV-infected cats when compared to uninfected cats. Additionally, by examining all nodes in the body, a regionally severe lymphadenopathy in FIV-infected cats was evident involving the lymph nodes of the hindlimb, forelimb, and head, in decreasing order of severity, with little evidence of enlargement in lymph nodes of the alimentary tract. Use of 99% confidence intervals showed that 9/9 FIV infected cats had enlarged lymph nodes of the hindlimb and forelimb region. In contrast, 7/9 and 3/9 FIV-infected cats exhibited enlargement of the nodes of the head region and alimentary tract, respectively. Similarly the combined weights of both left and right popliteal lymph nodes were enlarged in 9/9 FIV-infected cats whereas 0/6 in uninfected cats were not. The enlargement of the popliteal lymph nodes observed at necropsy was reflected microscopically by an increase in the size and number of germinal centres and an increase in the number of plasma cells, especially in the medullary cords. Because of the regional variation in lymph node size and numbers, it is suggested that the popliteal lymph node is a good indicator node for the assessment of lymph node status in FIV infection.

Animals↗
PubMed1995

Evaluation of an autologous red cell agglutination test, VetRED FIV, for the presence of FIV antibody in cats.

A commercially available whole blood agglutination test, VetRED FIV, used for the detection of antibodies to feline immunodeficiency virus (FIV), was evaluated. The test is based on the use of a synthetic peptide conjugated to a non-agglutinating anti-feline red blood cell monoclonal antibody. The amino acid sequence of the synthetic peptide was derived from the predicted sequence of the transmembrane protein of FIV. The sensitivity and specificity of VetRED FIV was 100% when 34 known FIV-positive and 15 known FIV-negative cats were tested. These cats were part of studies on experimentally induced FIV infection, with their FIV status confirmed by virus isolation. Further, VetRED FIV was compared with another commercially available test for FIV antibody, PetChek in a field trial on 548 feline blood samples received by a diagnostic laboratory. Of the test results 94.2% (516/548) were in agreement: 112 were positive by VetRED FIV and PetChek; 404 were negative by both tests and 32 were discordant. These 5.8% discordant samples producing VetRED FIV-positive/PetChek-negative or VetRED FIV-negative/PetChek-positive were further assessed by Western blot assay. In the field trial, the sensitivity and specificity of VetRED FIV was 97% and 97%, respectively, comparable to the 98% sensitivity and 99% specificity for PetChek. The results from the trial also confirm the relatively high overall prevalence of FIV in Australian cats predominantly among mature male cats in the 9-12 year age group. Given the simplicity of the VetRED FIV procedure, it is concluded that VetRED FIV is a useful addition to the available commercial tests for FIV infection.

Animals↗
PubMed1995

Feline immunodeficiency virus replicates in salivary gland ductular epithelium during the initial phase of infection.

Feline immunodeficiency virus (FIV) antigen was detected by immunochemistry in salivary glands of cats experimentally inoculated with West Australian isolate T91. Six cats were inoculated subcutaneously with 1.0 ml of tissue culture supernatant fluid from a feline T-lymphoblastoid cell line (MYA-1) infected with T91. FIV antigens were detected in the interlobular ducts of the salivary gland of cats infected with FIV 2, 4 and 6 weeks previously. FIV antigen was not detected in the salivary glands of three FIV negative cats and one naturally infected cat. Further, FIV antigen was located only in interlobular duct epithelial cells. The distribution of FIV in the interlobular ducts confirms the important role of salivary glands as a major reservoir of FIV in the early phase of infection and strengthens suggestions that the salivary route is an important mode of transmission of FIV.

Animals↗
PubMed1995

Bull terrier hereditary nephritis: a model for autosomal dominant Alport syndrome.

Bull terrier hereditary nephritis is inherited as an autosomal dominant disease and causes renal failure at variable ages in affected dogs. The aims of this study were to compare the clinical, ultrastructural and immunohistochemical features of bull terrier hereditary nephritis with the characteristics of the human forms of Alport syndrome. Many animals with bull terrier hereditary nephritis have hematuria, and some have anterior lenticonus. However, deafness is not associated with the renal disease, and affected dogs do not have the large platelets that are occasionally seen in patients with autosomal Alport syndrome. The glomerular capillary basement membrane (GCBM) in affected bull terriers has an identical ultrastructural appearance to that seen in X-linked Alport syndrome, with lamellations and intramembranous electron-dense deposits. However, both the Goodpasture and the Alport antigens, which represent parts of the alpha 3(IV) and alpha 5(IV) collagen chains, respectively, are present in the GCBM of affected dogs. Bull terrier hereditary nephritis represents an animal model for autosomal dominant Alport syndrome, and can be used to further examine how genetic mutations affect a basement membrane protein and the corresponding membrane structure.

Animals↗