Hyperplastic cholangitis in a naturally Toxoplasma gondii-infected cat.

Toxoplasma gondii can cause in cats a multisystemic disease involving the liver, lungs, central nervous system and other organs. The liver generally shows multifocal necrotizing hepatitis with possible panlobular extension, with histological evidence of free tachyzoites and/or cysts containing bradyzoites within necrotic foci. Very rarely, toxoplasmosis is expressed by cholangitis, the latter being much more frequently caused in cats by bacteria of intestinal origin. We report here a case of cholangitis/cholangiohepatitis in a young cat, where cytology of the liver showed multiple maturational stages of protozoa in the cytoplasm of cells of the bile ducts. On the basis of the cytological, histological, ultrastructural and molecular details, the microorganisms were identified as belonging to the species Toxoplasma gondii

Toxoplasmosi biliare in un gatto

Toxoplasma gondii pu\uf2 provocare nel gatto una malattia multisistemica coinvolgente fegato, polmoni, sistema nervoso centrale e altri organi. Nel fegato si osserva generalmente una epatite necrotizzante multifocale a possibile estensione panlobulare, con evidenza istologica di tachizoiti liberi e/o di cisti contenenti bradizoiti nel contesto dei focolai necrotici. Molto pi\uf9 raramente la toxoplasmosi si esprime con una colangite, che nel gatto \ue8 solitamente causata da germi di provenienza intestinale. Si presenta un caso di colangite/colangioepatite in un giovane gatto con ittero, vomito e diarrea, in cui l\u2019esame citologico del fegato ha evidenziato molteplici stadi maturativi protozoari nel citoplasma delle cellule delle vie biliari. Sulla base delle caratteristiche istologiche e citologiche e di approfondimenti ultrastrutturali e molecolari, i microrganismi sono stati identificati come appartenenti alla specie Toxoplasma gondii

Reduced diversity of immunoglobulin and T-cell receptor gene rearrangements in chronic inflammatory gastrointestinal diseases in dogs.

Abstract Inflammatory bowel disease has a multifactorial etiology in dogs as it does in humans. Evidence has been accumulated showing an abnormal response of the immune system, mostly represented by lymphocyte infiltration in the lamina propria of the gastrointestinal tract and in the epithelium, likely driven by chronic antigenic stimulation against luminal microorganisms. A relevant role is also ascribed to the genetic predisposition typical of some canine breeds. The role of chronic antigenic stimulation is still under debate. It may be responsible for selective pressure on the lymphoid population, favouring the emergence of some lymphocyte clones. This cross-sectional study is aimed at investigating the immunoglobulin and T-cell receptor gene rearrangements in a group of dogs affected by inflammatory bowel disease. The database of a referral Veterinary Laboratory was investigated. Based upon the histological evaluation of the bioptic samples collected during endoscopy, 54 canine cases met the WSAVA criteria for diagnosing IBD and were included in the study. The histological slides were retrieved and the gDNA was purified using protocols for formalin-fixed tissue. The gDNA was PCR amplified using fluorescent-labelled primers specific for canine immunoglobulin and T-cell receptor gene rearrangements; the PCR products were analysed with fragment analysis by means of capillary electrophoresis on an automatic sequencer (GeneScanning). In 47/54 (87.3%) cases, it was possible to amplify the gDNA. Twenty-one patients out of 47 (44.7%) showed polyclonal patterns in both the immunoglobulin and the T-cell receptors, 18/47 (38.3%) showed at least one oligoclonal pattern without monoclonal ones while 8/47 (17.0%) cases showed an Ig (7/47; 14.9%) or TCR (1/47: 2.1%) monoclonal pattern. These findings indicate that reduced diversity of the immunoglobulin and T-cell receptor repertoire occurs in canine inflammatory bowel disease. The reduced diversity correlated significantly with the severity of the histological lesions and carried a significantly increased risk of death. Beside its possible role as a reliable ancillary assay, immunoglobulin and T-cell receptor GeneScanning analysis points to the possible role of aberrant chronic antigenic stimulation, leading to clonal expansion of certain lymphocyte subsets in the pathogenesis of canine IBD

Recruited leukocytes and local synthesis account for increased matrix metalloproteinase-9 activity in cerebrospinal fluid of dogs with central nervous system neoplasm.

Matrix metalloproteinases (MMP) 2 and 9 are enzymes known to degrade several protein components of the extracellular matrix. In humans, increased concentrations of these enzymes have been demonstrated in the cerebrospinal fluid (CSF) of subjects affected by many neurological conditions including brain tumours; nevertheless comparative data in dogs are completely lacking. Aim of this study was to investigate these molecules in CSF of dogs diagnosed with CNS neurological diseases. Higher activity of MMP 2 and 9 was revealed in dogs with space occupying lesions of likely neoplastic origin in comparison to dogs with idiopathic epilepsy. Statistical modelling reveals that increased MMP 9 activity is mainly due to leukocytes recruitment and local synthesis

Gelatinases, endonuclease and Vascular Endothelial Growth Factor during development and regression of swine luteal tissue.

Background: The development and regression of corpus luteum (CL) is characterized by an intense angiogenesis and angioregression accompanied by luteal tissue and extracellular matrix (ECM) remodelling. Vascular Endothelial Growth Factor (VEGF) is the main regulator of angiogenesis, promoting endothelial cell mitosis and differentiation. After the formation of neovascular tubes, the remodelling of ECM is essential for the correct development of CL, particularly by the action of specific class of proteolytic enzymes known as matrix metalloproteinases (MMPs). During luteal regression, characterized by an apoptotic process and successively by an intense ECM and luteal degradation, the activation of Ca++/Mg++-dependent endonucleases and MMPs activity are required. The levels of expression and activity of VEGF, MMP-2 and -9, and Ca++/Mg++-dependent endonucleases throughout the oestrous cycle and at pregnancy were analyzed. Results: Different patterns of VEGF, MMPs and Ca++/Mg++-dependent endonuclease were observed in swine CL during different luteal phases and at pregnancy. Immediately after ovulation, the highest levels of VEGF mRNA/protein and MMP-9 activity were detected. On days 5\u201314 after ovulation, VEGF expression and MMP-2 and -9 activities are at basal levels, while Ca++/Mg++-dependent endonuclease levels increased significantly in relation to day 1. Only at luteolysis (day 17), Ca++/Mg++-dependent endonuclease and MMP-2 spontaneous activity increased significantly. At pregnancy, high levels of MMP-9 and VEGF were observed. Conclusion: Our findings, obtained from a precisely controlled in vivo model of CL development and regression, allow us to determine relationships among VEGF, MMPs and endonucleases during angiogenesis and angioregression. Thus, CL provides a very interesting model for studying factors involved in vascular remodelling

Evaluation of matrix metalloproteinases 2 and 9 activity in cerebrospinal fluid of dogs with non-inflammatory diseases of the central nervous system.

Matrix metalloproteinases (MMP) 2 and 9 are enzymes involved in extracellular matrix (ECM) degradation with specific proteolytic activity for type IV collagen and gelatin.MMPs participate in several physiological processes characterised by ECM remodelling and are implicated in various inflammatory, degenerative and neoplastic pathological conditions. The invasive and metastatic potential of several neoplastic lesions in the central nervous system (CNS) has been correlated with MMP expression; therefore, MMP assessment in cerebrospinal fluid (CSF) has gained an important diagnostic and prognostic role in human medicine (Friedberg et al., 1998).MMP2has been evidenced in the CSF of clinically normal dogs; nevertheless, MMP expression has not been evaluated in the CSF of dogs affected by pathological conditions of the CNS (Bergman et al., 2002). The aim of the present work was to retrospectively evaluate, using gelatin-zymography, the MMP2 and 9 activities in CSF samples obtained from dogs with non-inflammatory diseases of the CNS, with particular attention to space-occupying lesions (SOL). This last activity was evaluated in comparison with two other different pathological conditions: idiopathic epilepsy (IE) and disk herniation (DH)

Relationships between innovative and traditional parameters to investigate semen quality in pigs.

Semen quality assessment represents a fundamental step for obtaining successful artificial insemination (AI). In commercial settings, the semen employed for AI should be of high quality but traditional semen quality estimates are not sufficiently sensitive to discriminate between differences among samples in terms of fertilising ability. Therefore, more discriminative sperm characteristics need to be identified in order to better predict fertility outcome. In the present study, a series of molecular aspects of semen, represented by heat shock proteins, oxidative stress status, antioxidant potential and tumour necrosis factor (TNF)alpha were evaluated and analysed. Several relationships between traditional and investigated molecular semen quality estimates were found by using a multivariate analysis approach. Tumour necrosis factor (TNF)-alpha was identified in boar seminal plasma resulting in positive correlations with several sperm quality aspects and particularly with motility. The protective roles of antioxidant molecules and heat shock proteins have been demonstrated confirming the data previously published in the literature

KIT receptor dyregulations in feline mast cell tumours and systemic mastocytosis

none6Introduction: Feline mast cell tumours (FeMCTs), overall Accounting for 1e9% of feline neoplasms, are characterized by a highly variable biological behaviour. Frequent post- surgical recurrence, de-novo development of multiple tumours and concurrent visceral and cutaneous involvement justify uncertainty in differentiating benign from malignant forms, with tendency to systemic spread. Materials and Methods: A series of FeMCTs with variable clinical presentation were examined by histology (cell morphology, differentiation, growth pattern, mitotic activity), CD117 immunohistochemistry and c-Kit mutation analysis (exons 8, 9 and 11). Data were correlated with clinical records (clinical signs, TNM stage, haematological abnormalities and 2-year follow-up) to assess their prognostic significance. Results: Twenty cats with 10 solitary cutaneous, five multiple cutaneous and five systemic MCTs were included; nine cats were still alive at the end of the follow-up period. Overall, 29 tumour samples were examined. There were 21 well-differentiated, three pleomorphic and five atypical FeMCTs; mean mitotic activity was 9/10 high power fields. Low to high CD117 expression was observed in 18 cases. Further c-Kit mutations, beside those previously described in FeMCT, were found. Conclusions: This study investigated the effects of c-Kit dysregulation on FeMCT biological behaviour and also potentially allowed the identification of those patients that may benefit from molecular targeted therapies.mixedSabattini S; Guadagni-Frizzon M; Turba ME; Gentilini F; Capitani O; Bettini GSabattini S; Guadagni-Frizzon M; Turba ME; Gentilini F; Capitani O; Bettini

KIT receptor dyregulations in feline mast cell tumours and systemic mastocytosis.

Introduction: Feline mast cell tumours (FeMCTs), overall Accounting for 1e9% of feline neoplasms, are characterized by a highly variable biological behaviour. Frequent post- surgical recurrence, de-novo development of multiple tumours and concurrent visceral and cutaneous involvement justify uncertainty in differentiating benign from malignant forms, with tendency to systemic spread. Materials and Methods: A series of FeMCTs with variable clinical presentation were examined by histology (cell morphology, differentiation, growth pattern, mitotic activity), CD117 immunohistochemistry and c-Kit mutation analysis (exons 8, 9 and 11). Data were correlated with clinical records (clinical signs, TNM stage, haematological abnormalities and 2-year follow-up) to assess their prognostic significance. Results: Twenty cats with 10 solitary cutaneous, five multiple cutaneous and five systemic MCTs were included; nine cats were still alive at the end of the follow-up period. Overall, 29 tumour samples were examined. There were 21 well-differentiated, three pleomorphic and five atypical FeMCTs; mean mitotic activity was 9/10 high power fields. Low to high CD117 expression was observed in 18 cases. Further c-Kit mutations, beside those previously described in FeMCT, were found. Conclusions: This study investigated the effects of c-Kit dysregulation on FeMCT biological behaviour and also potentially allowed the identification of those patients that may benefit from molecular targeted therapies