Tissue and cellular characterisation of nucleolin in a murine model of corneal angiogenesis.

PURPOSE: Corneal neovascularisation (CNV), with consequent loss of transparency, is due to an imbalance of proangiogenic factors. Cell-surface nucleolin (NCL) has been associated with neo-angiogenesis. There are studies identifying NCL translocation from nucleus to the cell surface, which is essential for endothelial cell proliferation. To find the possible role of NCL in the generation of corneal neovessels, the aim of this study is to characterise the NCL presence and cell-localisation in non-injured corneas, as well as to describe the changes in NCL cell and tissue localisation in CNV, and to analyse the effect of bevacizumab on NCL cellular and tissular distribution. METHODS: Suture-induced CNV was performed in mice. The corneal tissues were obtained and the histological and co-immunofluorescence assays were performed using different proteins, such as CD31, cadherin and isolectin B4. To determine the possible role of VEGF in NCL presence and localisation in our CNV model, bevacizumab was concomitantly used. RESULTS: Nucleolin was principally observed in the nucleus of the basal epithelial cells of normal corneas. Interestingly, angiogenesis-induced changes were observed in the localisation of NCL, not only in tissue but also at the cellular level where NCL was extranuclear in epithelial cells, stromal cells and neovessels. In contrast, these changes were reverted when bevacizumab was used. Besides, NCL was able to stain only aberrant corneal neovessels in comparison with retinal vessels. CONCLUSIONS: NCL mobilisation outside the nucleus during angiogenesis could have a possible role as a proangiogenic molecule in the corneal tissue.

Comparative analysis of inflammatory response in the BALB/c and C57BL/6 mouse strains in an endotoxin-induced uveitis model.

Uveitis is an inflammatory disease associated with diverse systemic and autoimmune diseases, defined as the inflammation of any given segment of the uveal tract, uveitis contributes with 5-20% cases of blindness in the USA/Europe and >25% of cases in third-world countries. To understand its pathogenic mechanisms, BALB/c and C57BL/6 mice were induced to develop the condition by a single intraperitoneal injection of E. coli lipopolysaccharide, the aim of the present work is to determine leukocyte infiltration in an endotoxin-induced uveitis (EIU) in two non-related mouse strains. Histopathological findings and clinical analysis were conducted 24 and 48 h postinjection. Both strains presented conventional clinical signs of uveitis 24 h post LPS injection and the highest inflammatory leukocyte infiltration in the uveal tract was found in the BALB/c mouse strain. This article will give an insight on the difference of the inflammatory response in the EIU model in two different mouse strains and the relationship between the pathologic response.

Presentación sistémica de Aspergillus spp con semiología neurológica en un Pastor Alemán: informe de un caso clínico / Systemic presentation of Aspergillus spp with neurological involvement in a German Shepherd: clinical case report

Infections of the central nervous system are uncommon in dogs. Pathogenic fungi such as Aspergillus, Cryptococcus spp, Blastomyces dermatitidis and Coccidioides immitis have been reported. An Aspergillus infection is mainly a respiratory event and very seldom it may become disseminated, the German shepherd seems to be the more prone to this type of aspergillosis specially if an immunological deficiency is present. A 1.5 year old female German Shepherd from Cancun, Quintana Roo, State, in the southeast of Mexico was presented to the small animal hospital, she showed tetraparesis, hiporeflex and deep retarded sensitivity in all four members, loss of sensitivity in the left side of the face, absence of pupil reflex and menace reflex as well as absence of muscular tone of the vulva and anal sphincter. The third day after presentation, she died. At necropsy polyencephalomalace, mielitis and diffuse leucomielitis piogranomalotosus were found. In the cytology of lymph node and other tissues aspergilli like hiphae were observed, this was confirmed by using the highly specific double diffusion test. Then, a PCR test was carried out with tissue fixed in 10% formaldehyde, but it failed probably due to destruction of the DNA.

La infección del sistema nervioso en el perro es poco común; posibles causas son Aspergillus spp, Cryptococcus neoformans, Blastomyces dermatidis y Coccidioides immitis. La infección por Aspergillus spp afecta al aparato respiratorio y en pocas ocasiones se presenta de manera diseminada. Sin embargo, en el Pastor Alemán hay mayor incidencia de éste, pues dicha infección se asocia con una deficiencia inmunológica. Se describe el caso de una perra de raza Pastor Alemán, de 1.5 años de edad, referida de Cancún, Quintana Roo, México, con tetraparesis, hiporreflexia y sensibilidad profunda retardada en los cuatro miembros, disminución de sensibilidad en el lado derecho de la cara y ausencia de reflejo pupilar y de amenaza en ojo derecho; además de ausencia de tono vulvar y de esfínter anal. A los tres días después del ingreso la perra falleció. A la necropsia se encontró polioencefalomalacia, mielitis y leucomielitis piogranulomatosa difusa. En la citología de linfonodo y en la histopatología de diferentes tejidos se identificaron hifas compatibles con Aspergillus spp. Con el fin de dar certeza al diagnóstico, se realizó prueba de doble inmunodifusión, que es altamente específica, la cual confirmó el diagnóstico de Aspergillus spp. Posteriormente se corrió PCR a partir de una muestra de tejido en formalina al 10%, para identificar la especie, en donde no se lograron resultados, probablemente debido a la destrucción del ADN por el formaldehído.