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Biomedical subjects

D Fondevila

Publications and source records attributed to D Fondevila.

At least 19 recordsLinked to original sources

Evaluation of the expression of P-selectin, ICAM-1, and TNF-alpha in bacteria-free lesional skin of atopic dogs with low-to-mild inflammation.

Canine atopic dermatitis (AD) is a pruritic skin condition that shares many clinical and pathophysiological features with its human counterpart. A major therapeutic challenge of AD is the control of the skin inflammatory process. A detailed knowledge of the pro-inflammatory molecules involved in cell recruitment in AD would allow for a better control of the disease. We thus have studied the protein expression of P-selectin, ICAM-1 and TNF-alpha in the lesional and non-lesional skin of atopic dogs that had been treated for bacterial infections. Despite a low-to-mild inflammatory process, P-selectin protein was clearly upregulated in the lesional skin areas when compared with non-lesional skin (four-fold average increase). This P-selectin upregulation was accompanied by signs of functional changes such as increased cell margination, and membrane-associated protein expression. Although the expression of ICAM-1 and TNF-alpha was not enhanced in the lesional versus the non-lesional skin, there was a trend towards a correlated upregulation of both molecules. Further studies will help elucidate the significance of the substantial overexpression of P-selectin in canine AD, in particular in a scenario where bacterial antigens are not contributing as pro-inflammatory stimuli.

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PCR technique detection of Leishmania spp. but not Mycobacterium spp. in canine cutaneous 'sterile' pyogranuloma/granuloma syndrome.

Cutaneous 'sterile' pyogranuloma/granuloma syndrome (SPGS) is an uncommon canine skin disorder of unknown aetiopathogenesis. Histopathological findings and failure to demonstrate an aetiologic agent are suggestive of this syndrome. Nevertheless, it has been hypothesized that SPGS may be related to an immune response against persistent endogenous or exogenous antigens. The presence of Leishmania and Mycobacterium organisms was investigated by polymerase chain reaction (PCR) techniques in 46 canine skin samples histopathologically diagnosed as SPGS. Concomitantly, an immunohistochemical technique for Leishmania detection was applied on the same samples and the results were compared with those from PCR. The PCR technique yielded positive results for Leishmania spp. in 21 out of 46 skin samples. The results of immunohistochemical techniques were identical to those obtained by PCR. The PCR technique gave negative results for Mycobacterium spp. in all the samples examined. These results suggest the importance of looking for Leishmania spp. in skin biopsies with histopathological findings consistent with the diagnosis of SPGS.

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Histological and immunohistochemical study of clinically normal skin of Leishmania infantum-infected dogs.

Skin lesions are the most usual manifestation of canine leishmaniosis. The aim of this study was to investigate the histological pattern and parasite load in clinically normal skin of Leishmania-infected dogs. Two groups of Leishmania-infected dogs were studied. Group A consisted of 15 symptomless animals which, although seronegative or only mildly seropositive, gave a positive polymerase chain reaction (PCR) for Leishmania in the skin. Group B consisted of 20 clinically affected dogs which were highly seropositive and PCR-positive. Biopsies of normal skin from all dogs were processed for routine histology and Leishmania immunohistochemistry. The study demonstrated microscopical lesions and the presence of parasites in the skin from dogs of group B, but not group A. The results cast doubt on the relevance of infected but symptomless dogs in the epidemiology of canine leishmaniosis. In contrast, however, the clinically normal skin of sick dogs harbours the parasite and probably plays a role in the transmission of leishmaniosis.

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Piecemeal degranulation (PMD) morphology in feline circulating eosinophils.

Buffy coat preparation from six cats with 600-4560 circulating eosinophils/microL was collected by either blood centrifugation or sedimentation, fixed in 2.5% glutaraldehyde, post-fixed in either 1% osmium or in 1.5% potassium ferrocyanide-reduced osmium, ultra-sectioned and examined by transmission electron microscopy. Ultrastructural changes of piecemeal degranulation (PMD), which is a mechanism of eosinophil granule contents release indicative of eosinophil activation, were observed in specific granules from all the samples examined. The spectrum of PMD included coarsening of the granule matrix, budding vesicles, fragmented granule cores and lucent granules. The number of presumably activated eosinophils with ultrastructural evidence of PMD did not correlate with the level of eosinophilia. The lack of correlation suggested that, analogously with humans, blood eosinophil count might not represent the best criterion to evaluate the contribution of eosinophils to tissue damage in certain feline eosinophil-associated diseases.

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Histopathological study of feline eosinophilic dermatoses.

A retrospective study was conducted on skin specimens from 24 cats with eosinophilic granuloma complex. The specimens were stained with haematoxylin and eosin and Gallego's trichrome stain. In all specimens, flame figures and/or large foci of so-called "collagen degeneration" were detected and histopathological features were not predictive of the clinical picture. Use of the term eosinophilic dermatosis was advocated in diagnostic dermatopathology. On trichrome-stained sections, normally stained collagen fibres were identified in the middle of both flame figures and large foci of "collagen degeneration" and the debris surrounding collagen bundles showed the same tinctorial properties as eosinophil granules. Eosinophil degranulation around collagen bundles seemed to represent the major pathogenetic event in these lesions, analogous with human flame figures. The term flame figures might therefore be more accurately used to designate those foci of eosinophilic to partly basophilic debris commonly referred to as "collagen degeneration".

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Assessment of proliferative activity of canine dermal mast cells by bromodeoxyuridine and proliferating cell nuclear antigen labelling.

Cutaneous mast cells from skin biopsies of three healthy dogs and three dogs with atopic dermatitis were assessed for their proliferative potential using bromodeoxyuridine and proliferating cell nuclear antigen labelling. Mast cells isolated from the skin of two healthy dogs were also studied using bromodeoxyuridine labelling. Mast cells in skin biopsy specimens and mast cells isolated from the skin of healthy dogs did not incorporate bromodeoxyuridine. Two mast cells expressing proliferating cell nuclear antigen were seen around two superficial vessels in the dermis of one atopic dog. Epidermal cells, glandular epithelial cells, fibroblasts and endothelial cells incorporated bromodeoxyuridine and showed positive staining for proliferating cell nuclear antigen. These results suggest that canine mature mast cells do not proliferate in the dermis.

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A case of feline phaeohyphomycosis due to Fonsecaea pedrosoi.

The first report of a case of feline phaeohyphomycosis due to Fonsecaea pedrosoi is presented. Fonsecaea pedrosoi is an aetiologic agent of both human phaeohyphomycosis and chromoblastomycosis. In our cat, the lesion was confined to the skin and appeared as a firm swelling on the bridge of the nose. Diagnosis was based on histological examination of a cutaneous biopsy and fungal culture of a tissue sample on Sabouraud's dextrose agar. Further diagnostic tests failed to reveal an underlying immunosuppression. Two treatment cycles with itraconazole, at the oral dose of 5 mg kg-1 given twice daily, induced complete clinical remission, but relapses occurred.

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Neuronal vacuolation in young Rottweiler dogs.

Neuronal vacuolation, involving the cerebellar roof nuclei, Purkinje cells, selected nuclei of the brain stem, thalamus, Clarke's column, anterior and posterior horns of the spinal cord, visceral autonomic ganglia and myenteric plexus, as well as axonal degeneration of the white matter of the brain stem, cerebellar pedunculi, dorsolateral columns of the spinal cord and ventral roots of the spinal cord, were observed in two young Rottweiler dogs which were clinically afflicted with hind limb weakness progressing to paraparesia, ataxia, intention tremor, and difficulty in swallowing and barking. The absence of modifications in Bcl-2 and Bax immunoreactivity, a lack of strong c-Jun/AP-1 (N) immunoreactivity in vacuolated cells, and the absence of DNA breaks, as seen with the method of in situ end-labeling of nuclear DNA fragmentation, all suggest that there is no involvement of the apoptotic pathway in vacuolated cells in this new neurodegenerative disorder.

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Comparative immunohistochemical study of stellate cells in normal canine and equine adenohypophyses and in pituitary tumours.

The presence and distribution of S100 protein (alpha and beta subunits), cytokeratin polypeptides, glial fibrillary acidic protein, neurofilaments, vimentin, neuron specific enolase, synaptophysin, HLA class II DR antigen, and pituitary hormones (prolactin, adrenocorticotropic hormone and human chorionic gonadotrophin) in stellate cells were studied immunohistochemically in four normal canine pituitary glands, five canine pituitary adenomas, two canine pituitary carcinomas and two equine pituitary adenomas (with surrounding normal glandular tissue). Stellate cells of the pars distalis and pars intermedia of canine and equine adenohypophyses showed a strong reaction with antibodies against S100 protein subunits alpha and beta. They also reacted with antibody against high and low molecular weight cytokeratins, but not with those against other intermediate filament proteins, neuroendocrine markers, the HLA-class II DR antigen or the pituitary hormones. Other populations of cells expressing both subunits of the S100 protein were polygonal cells of the pars distalis of the adenohypophysis (horse) and marginal epithelial cells of the pars intermedia of the adenohypophysis (dog and horse). Some pituitary tumours had S100-immunoreactive cells with a distribution of alpha and beta subunits that differed between the two species. Some canine tumours (one adenoma and one carcinoma) expressed only the alpha subunit, but both of the equine adenomas expressed alpha and beta protein subunits. Some of the S100-immunoreactive tumour cells reacted with RCK-102 (cytokeratins 5+8) antibody in the dog but not in the horse. The results suggested that canine and equine stellate cells of the adenohypophysis are more closely related to epithelial than to glial cells, as is the case in cattle, sheep and goats but not human beings or mice. No subpopulation of cells of bone marrow origin could be identified among canine stellate cells, as they lack MHC class II antigen. The results also suggested that the presence of S100-immunoreactive cells is more striking in canine and equine tumours than in human tumours.

Adenoma↗

Primary central nervous system T-cell lymphoma in a cat.

A 5-year-old intact male Persian cat was presented with progressive neurologic deficits, hyperesthesia, hyperreflexia, ataxia, and intention tremors, which resulted in death. Serologic tests for feline leukemia virus and feline immunodeficiency virus infection were negative. Neurohistologic examination revealed the presence of pleomorphic cellular infiltrates in cerebral leptomeninges and around parenchymal vessels. The majority of infiltrating cells were uniformly immunostained using an antiserum directed against T cells. Immunohistochemical examination of paraffin-embedded brain sections for the antigens of canine distemper virus, herpesvirus, Borna virus, and Toxoplasma gondii were negative. Histologic and immunohistochemical studies revealed a primary central nervous system T-cell lymphoma affecting the brain.

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Epidermal immunocompetence in canine leishmaniasis.

The antigen-presenting cell function of the epidermis was investigated immunocytochemically in 16 dogs with different types of skin lesions induced by Leishmania infection. The degree of epidermal immunocompetence was evaluated according to the presence of Langerhans cells (LC) and keratinocytes expressing class II major histocompatibility complex (MHC II) molecules on the one hand, and the relative numbers of macrophages, T cells, and parasites in the dermis on the other, as described for human cutaneous leishmaniasis. In alopecic dermatitis, appropriate numbers of LC and MHC II-positive keratinocytes were shown to be associated with a mild T cell infiltration without significant numbers of parasites. By contrast, when the epidermis lacked antigen-presenting cells, as occurred in nodular lesions, macrophages and parasites massively infiltrated the dermis. Ulcerative lesions showed intermediate patterns of inflammation. These results suggest that dogs with alopecic dermatitis develop an effective control of the infection, whereas those with a generalized nodular disease mount an impaired immune response against the parasite. Skin lesions in dogs infected by Leishmania might not only have a prognostic value, but also represent a suitable model to study the natural course of human cutaneous leishmaniasis.

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Immunocytochemical study of the pathogenesis of Pacheco's parrot disease in budgerigars.

Thirty-five budgerigars were infected with the Psittacid herpesvirus 1 (RSL-1 strain, ATCC) to study the pathogenesis of Pacheco's disease. Intramuscular (i.m.) and oral (p.o.) infection routes were used in 21 and 14 animals respectively. Animals were euthanized on days 1, 2, 3, 4, 6 and 8 post-inoculation (p.i.) and complete postmortem examinations and histological studies were performed. The presence of viral antigen in tissues was detected by immunohistochemical techniques using a rabbit polyclonal antibody. In the i.m.-infected birds, lesions were first detected on the third day p.i. and included necrotizing hepatitis and splenitis, both associated to the presence of viral antigen. Necrotic and inflammatory lesions as well as viral antigen were detected in many organs after the fourth day p.i. (oesophagus, crop, pancreas, kidney, adrenal gland, thyroid and parathyroid glands, thymus, ovary and feathers) proving generalization of the disease. Chronology of the infection was similar in the p.o.-infected birds. However, two main differences were observed between the groups: In the p.o. group, viral antigen was first detected in cloacal mucosa (3rd day p.i.), liver and spleen; and viral entry into target cells and dissemination to the rest of the tissues was slower in this group. In addition, detection of viral antigen in feather follicular epithelial cells and in granulosa cells of organ follicles are findings that could be of relevance to the transmission of the virus.

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Immunohistochemical detection of CD31 antigen in normal and neoplastic canine endothelial cells.

This report describes the immunohistochemical detection of von Willebrand's factor (vWf) and CD31 antigen in paraffin wax-embedded, formalin-fixed tissue sections of canine normal organs and vascular neoplasms. CD31 antigen and vWf were detected in endothelial cells of all organs examined, except for the endothelia of the renal glomeruli, which were negative for vWf. All haemangiomas examined (15) were positive for both markers. Eleven of 15 haemangiosarcomas were positive for vWf and all 15 expressed the CD31 antigen. All other neoplasms investigated (fibrosarcomas, schwannomas, haemangiopericytomas) were negative for both markers. It is concluded that immunohistochemical detection of CD31 antigen is of value for studying vascular disorders of the dog in routinely processed tissue.

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