196,123 research outputs found
Production and characterization of a polyclonal antibody against the bovine prion-like doppel protein
No full textPRODUCTION AND CHARACTERIZATION OF A POLYCLONAL ANTIBODY AGAINST THE BOVINE PRION-LIKE DOPPEL PROTEIN
Marco Rondena, Fabrizio Ceciliani, Chiara Bazzocchi, Saverio Paltrinieri
Dipartimento di Patologia Animale, Igiene e Sanità Pubblica Veterinaria – Milano
AIM OF THE STUDY: Doppel (Dpl) is a prion-like protein recently identified in mammalian. The sequence is notably homologous to that of PrP. Dpl is associated to neurodegeneration when ectopically expressed in transgenic mouse. The aim of the study is to raise and characterize a polyclonal antibody (pAb) against bovine Dpl to study the tissue distribution of this protein. MATERIAL AND METHODS. A synthetic peptide spanning from residues 67 to 81 of bovine Dpl sequence was synthetized. Using this peptide, a pAb (Dpl 67-81), was raised in rabbit and purified by means of Protein A affinity chromatography. In order to test the specificity of Dpl 67-81, bovine Dpl was cloned and overexpressed in E.coli in the expression vector PQE30 and purified following methal chelate affinity and bovine PrP was purified by means of cation-exchange chromatography and cobalt-chelate affinity chromatography. Homogenate of tissues were tested for the presence of Dpl after SDS-PAGE and Western Blotting. Blots were visualized using ECL. Immunohistochemistry on cryostatic sections using the avidin-biotin complex methods was performed. RESULTS: Dpl 67-81 strongly react against rDpl (titre =1:20000) and not against bovine PrP. Dpl was detected in testis, spleen, limphnodes by both immunoblotting and immunohistochemistry. In particular endothelial and stromal cells showed a granular cytoplasmic positivity. CONCLUSIONS: Dpl 67-81 can detect the expression of Dpl in tissues using both Western Blotting and immunohistochemistry. Moreover, it does not crossreact with PrP, and therefore it could be also used in experiments of functional blocking of Dpl in order to study its function and its relationship with PrP
Eugenio Corti e Primo Levi a confronto: la parola come ribellione al sistema totalitario
No full textIl filosofo Aristotele, nella Politica, afferma: «la parola è fatta per esprimere ciò che è giovevole e ciò che è nocivo e, di conseguenza, il giusto e l’ingiusto». Di fronte alle nefandezze del «secolo breve», così come Hobsbawm definisce il Novecento, il potere della parola acquista un valore fondamentale per poter giudicare quanto è accaduto. In modo particolare la lezione di Primo Levi, deportato ad Auschwitz sotto il nazifacismo e quella di Eugenio Corti, sottotenente del 35° corpo d’armata che partecipò alla ritirata di Russia, in modo simile e al contempo diverso, mostrano, con le loro opere letterarie, quanto la parola sia la risposta al potere totalitario: un contropotere all’ideologia nazista e comunista. Si vogliono quindi confrontare queste due esperienze mettendo in luce quanto la letteratura riesca a far crollare poteri egemonici basati sulla distruzione della persona e sul convincimento di un mondo svincolato dalle libertà di pensiero
Immunohistochemical characterization of the leucocytic infiltrate associated with canine seminomas
No full textBoth canine and human seminomas are typically associated with leucocytic infiltration, the role of which remains poorly understood. In this study, leucocytes infiltrating 10 canine seminomas were characterized. Monoclonal antibodies directed against CD18, CD11a, CD11b, CD11c, CD21, CD3, CD4, CD8 and Major Histocompatibility Complex class I and II (MHC I and MHC II) were also employed. Infiltrating leucocytes were located around vessels, adjacent to the thin fibrous septa between neoplastic lobules, and were also scattered singly amongst neoplastic cells. The more profuse infiltrates often had the appearance of follicular aggregates. Immunohistochemically, all the samples showed generally similar results. Most of the infiltrating cells were positive for CD18 and CD11a. Infiltrating cells were mainly T lymphocytes (CD3þ), particularly of the CD8þ subset. B lymphocytes (CD21þ) were detectable in almost all infiltrates; in the follicular aggregates they were centrally located, whereas T lymphocytes (CD3þ) lined the periphery. CD11cþ cells were always more numerous than CD11bþ cells, demonstrating that if macrophages and antigen-presenting cells (APCs) were well represented, monocytes and granulocytes were practically absent. Almost all of the infiltrating cells were positive for both MHC I and MHC II antigens and, in nine samples, a large number of neoplastic cells expressing MHC I were detected. A few neoplastic cells expressing MHC II were observed in seven cases. The presence of CD8þ lymphocytes, together with the large number of both infiltrating and neoplastic cells expressing MHC I, suggests that the lymphocytes mediate a cytotoxic reaction against the neoplastic cells. This hypothesis may underlie the favourable prognosis frequently associated with canine seminomas
Classical and spermatocytic seminoma in the dog : histochemical and immunohistochemical findings
No full textIn the light of earlier human studies, 43 canine tumours diagnosed as seminoma were examined histologically with haematoxylin and eosin and periodic acid-Schiff (PAS) stains, and immunohistochemically with a monoclonal antibody against human placental alkaline phosphatase (PLAP). Twenty tumours were positive for both PAS and PLAP and were therefore diagnosed as classical seminoma (SE). The other 23 tumours were negative for both PAS and PLAP and were therefore diagnosed as spermatocytic seminoma (SS). Tubules with carcinoma in situ (CIS) were present in the testicular parenchyma surrounding 15 SEs and nine SSs
A case of multiple corneal epithelial inclusion cysts in a dog
No full textA case of two corneal epithelial inclusion cysts affecting the left eye, in a 3 years old
female
Labrador cross was investigated. The etiopathogenesis of the cysts formation was
suspected to be
traumatic. The two masses were removed successfully by superficial keratectomy followed
by a
third eyelid flap. Histologically, both lesions were represented by cystic formations lined by
multilayered squamous epithelium, consistent with stromal inclusion of surface corneal
epithelium.
Recovery was complete and there was no recurrence at three months follow up
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