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Mammary tumors are a very common neoplasm in the dog and show histological features and biological behaviour similar to human mammary carcinomas. Recently, a pathway named Wnt-1, involving beta-catenin and APC protein, has emerged as an important player in many human tumor types, including mammary neoplasms.Thirty-five samples of canine mammary tumors (10 benign and 25 malignant) were studied in order to evaluate the co-expression of beta-catenin, APC protein and E cadherin with confocal laser microscopical observation, by western blot analysis and by correlating data obtained with the histological grade of tumours.A progressive decrease of E-cadherin together with disruption of beta-catenin expression was observed in less differentiated malignant tumors. In addition, a loss of beta-catenin membranous distribution and a cytoplasmic accumulation was often coexpressed with disrupted expression of APC protein. Western blot analysis showed a progressive increase of beta-catenin in malignant tumors, which could be the expression of disrupted /-catenin catabolism leading to cytoplasmic accumulation. In some less differentiated malignant tumors, a marked beta-catenin decrease was also observed. This feature could be linked to mutations in beta-catenin gene coding for a truncated and lighter protein.These results may indicate the multifunctional role played by beta-catenin in canine mammary oncogenesis.

The Red Palm Weevil (RPW) is one of the major pests of palms, frequently leading to the plants death. Action plans and the development of bio/physical strategies to contrast RPW diffusions are strongly recommended due to the serious concerns related to environmental pollution and insects’ resistance to chemicals. In the present study, we investigated morphological alterations of the ovaries and testes in adult RPW exposed to 2.45 GHz for 5, 15, and 30 s. During these treatments, the relative increase in temperature and the days of survival after irradiation were monitored. Then, RPWs were processed for macroscopical and microscopical analysis. Histological lesions of the ovaries and testes were characterized by the degeneration and necrosis of germinal cells, which increased with the increase in the time of irradiation and the temperature. By the same token, an increase in the temperature of irradiated insects was associated with a decrease in their survival time. These observations lead us to conclude that MWs could represent a useful tool for reducing or eliminating the reproductive capacity of this dreaded insect.

Recent reports of Eucoleus garfiai in wild boars in southern Italy have highlighted the need for collecting epidemiological data on the presence of this parasite and understanding the role of possible interactions between wild boars, E. garfiai, and the environment. This study analyses, using histopathological and biomolecular techniques, the presence of E. garfiai in tongue samples of wild boars hunted in four provinces of the Campania and Latium regions (Italy), in areas located above and below 900 m above sea level (asl). Histopathological examinations revealed the presence of adults and eggs of nematodes, which were subsequently identified as E. garfiai by biomolecular analysis, in the tongue epithelium. The detection of the parasite was more frequent in samples collected from hunting areas located above 900 m asl than in those collected from areas located below 900 m asl (66.67% vs. 38.09%; p < 0.01). Some species of earthworms are intermediate hosts of E. garfiai and it is well known that earthworms are more present in high quality soils. Therefore, we can suggest that the higher prevalence of E. garfiai at higher altitudes is probably linked to a greater presence of earthworms in the soil, due to its higher quality in these areas.

Simple Summary Equine sarcoids, caused by bovine papillomaviruses, are equine skin tumors of fibroblastic origin. It is well known that bovine papillomaviruses are able to interfere with the survival and proliferation of cells by regulating autophagy, a mechanism implicated in the breakdown and reuse of old and damaged cellular material. The present study focused on the evaluation in equine sarcoids and normal skins of the expression level of some of the main proteins involved in the autophagic pathway, such as Beclin 1, LC3 and P62, by immunohistochemical and biochemical techniques. Results obtained in equine sarcoids suggested an alteration of the autophagic process which could lead to a predominance of a particular population of fibroblast. Those fibroblasts could survive longer in a hypoxic microenvironment and produce more and/or altered collagen, giving an origin to the equine sarcoid. Abstract Background: It is well known that δ-bovine papillomaviruses (BPV-1, BPV-2 and BPV-13) are one of the major causative agents of equine sarcoids, the most common equine skin tumors. Different viruses, including papillomaviruses, evolved ingenious strategies to modulate autophagy, a complex process involved in degradation and recycling of old and damaged material. Methods: The aim of this study was to evaluate, by immunohistochemistry (IHC) and Western blot (WB) analysis, the expression of the main related autophagy proteins (Beclin 1, protein light chain 3 (LC3) and P62), in 35 BPV1/2 positive equine sarcoids and 5 BPV negative normal skin samples. Results: Sarcoid samples showed from strong-to-moderate cytoplasmic immunostaining, respectively, for Beclin 1 and P62 in >60% of neoplastic fibroblasts, while LC3 immunostaining was weak to moderate in ≤60% of neoplastic fibroblasts. Western blot analysis confirmed the specificity of the antibodies and revealed no activation of autophagic flux despite Beclin 1 overexpression in sarcoid samples. Conclusion: Results could suggest the activation of the initial phase of autophagy in equine sarcoids, and its impairment during the following steps. The impairment of autophagy could lead to a selection of a quiescent population of fibroblasts, which survive longer in a hypoxic microenvironment and produced more and/or altered collagen.

The Oriental hornet (Vespa orientalis) is spreading across the Italian territory threatening the health and wellbeing of honeybees by feeding on adult individuals and larvae and by plundering hive resources. Considering the capacity of other hornets in harboring honeybee viruses, the aim of this study was to identify the possible role of the Oriental hornet as a vector for honeybee viruses. Adult hornets were subjected to macroscopical examination to identify the presence of lesions, and to biomolecular investigation to detect the presence of six honeybee viruses: Acute Bee Paralysis Virus (ABPV), Black Queen Cell Virus (BQCV), Chronic Bee Paralysis Virus (CBPV), Deformed Wing Virus (DWV), Kashmir Bee Virus (KBV), Sac Brood Virus (SBV). No macroscopical alterations were found while biomolecular results showed that DWV was the most detected virus (25/30), followed by ABPV (19/30), BQCV (13/30), KBV (1/30) and SBV (1/30). No sample was found positive for CBPV. In 20/30 samples several co-infections were identified. The most frequent (17/30) was the association between DWV and ABPV, often associated to BQCV (9/17). One sample (1/30) showed the presence of four different viruses namely DWV, ABPV, BQCV and KBV. The detected viruses are the most widespread in apiaries across the Italian territory suggesting the possible passage from honeybees to V. orientalis, by predation of infected adult honeybees and larvae, and cannibalization of their carcasses. However, to date, it is still not clear if these viruses are replicative but we can suggest a role as mechanical vector of V. orientalis in spreading these viruses.

Papillomavirus (PV) infection is associated with development of epithelial cancer in different species, including domestic cat (Felis catus). Felis catus PV type-2 (FcaPV-2) is considered the causative agent of a proportion of feline cutaneous squamous cell carcinoma (SCC), through the transforming properties of its E6 and E7 oncogenes. However, the possible role of FcaPVs in the aetiology of feline oral SCC (FOSCC) is still unclear. The aim of this study was to assess the presence and gene expression of FcaPV-2 in FOSCC samples. We detected FcaPV-2 DNA in 10/32 (31%) of the analysed FOSCC by the use of PCR methods. Importantly, viral mRNA was detected by RT-PCR in 7/10 (70%) of DNA positive samples. In particular, FcaPV-2 L1, E2 and E6E7 genes were found to be expressed in 5/10 (50%), 3/10 (33%) and 5/10 (50%) samples, respectively. Viral DNA was also detected in non neoplastic oral ulcerative lesions (ULs) (4/11, 36%); qPCR suggested a difference in viral load between ULs and FOSCCs, particularly in those expressing E6E7, although it was not statistically significant. These data suggest, but do not definively prove, a possible role of FcaPV-2 in the development of a proportion of FOSCC. Moreover, L1 and E2 gene expression results indicate that FcaPV-2 infection associated with these tumours may possibly be productive.

Background: equine sarcoids are the most frequent skin tumors in equidae worldwide. It is well known that delta bovine papillomaviruses are their causative agents. We have recently shown the presence in equine sarcoids of abnormal vessel structures, which could cause a hypoxic condition. The aim of this study was to analyze the expression of hypoxia-inducible factor-1 alpha (HIF-1&alpha, ) in a subset of BPV positive equine sarcoids and explore the relationship with vascular endothelial growth factor (VEGF) expression. Results: 80% of equine sarcoids showed strong cytoplasmic staining in &gt, 60% of neoplastic fibroblasts, while 20% of samples showed a moderate cytoplasmic staining in 40&ndash, 60% of neoplastic fibroblasts for HIF-1&alpha, Results of Western blotting (WB) were consistent with immunohistochemistry (IHC). Moreover, a positive correlation between HIF-1&alpha, and VEGF expression (r = 0.60, p &lt, 0.01) was observed. Conclusion: we have shown that HIF-1&alpha, was strongly expressed in equine sarcoid. The upregulation of HIF-1&alpha, has been described in numerous tumors and can be modulated by many proteins encoded by transforming viruses. Thus, it is also possible that BPV could have a relevant role in HIF-1&alpha, pathway regulation, contributing to the development of equine sarcoids by promoting HIF-1&alpha, /VEGF mediated tumor angiogenesis.

Telomerase activity contributes to cell immortalization by avoiding telomere shortening at each cell division; indeed, its catalytic subunit telomerase reverse transcriptase (TERT) is overexpressed in many tumors, including human oral squamous cell carcinoma (hOSCC). In these tumors, matrix metalloproteinases (MMPs), a group of zinc-dependent endopeptidases involved in cell migration, contribute to invasive potential of cancer cells. A proportion of hOSCC is associated with infection by high-risk human papillomavirus (HR-HPVs), whose E6 oncogene enhances TERT and MMPs expression, thus promoting cancer progression. Feline oral squamous cell carcinoma (FOSCC) is a malignant tumor with highly invasive phenotype; however, studies on telomerase activity, TERT, and MMPs expression are scarce. In this study, we demonstrate telomerase activity, expression of TERT, and its transcriptional activator cMyc along with expression of MMP-1, -2, and -9 in FOSCC-derived cell lines SCCF2 and SCCF3, suggesting a contribution by these pathways in cell immortalization and invasion in these tumors. Recent studies suggest that a sub-group of FOSCC as well as SCCF2 and SCCF3 are associated with Felis catus PV type-2 (FcaPV-2) infection. However, in this work, FcaPV-2 E6 gene knock-down caused no shift in either TERT, cMyc, or MMPs levels, suggesting that, unlike its human counterpart, the viral oncogene plays no role in their regulation.

Canine mammary tumours are frequent neoplasms mostly affecting intact female dogs, for which no 100% efficient therapy is available. Platelet derived growth factor β receptor (PDGFβR) is a tyrosine kinase receptor (TKR) with a potential role in human breast cancer and a series of canine tumours. In this study we demonstrated, for the first time, expression of PDGFβR and its downstream transduction molecules, mitogen-activated protein/extracellular signal-regulated kinase kinase (MEK) and extracellular signal-regulated kinase (ERK), as well as their activated forms in canine mammary tumours by both biochemical analysis and immunohistochemistry. PDGFβR was expressed and hyperphosphorylated in the majority of tumour samples and tumour derived cell lines. Additionally, both MEK and ERK were expressed and activated in cell lines as well as biopsies. TKR inhibitors (TKRi) are currently under investigation as possible therapy in human breast and several canine tumours, thus our in vivo and in vitro findings pave the way for future studies aimed at establishing a potential therapeutic employment of TKRi for the treatment of canine mammary cancer.

Due to high rates of proliferation and DNA synthesis, neoplastic cells have higher requirements of iron than normal cells. For that reason, neoplastic cells have remodelled iron metabolism pathways, over‐expressing genes encoding for iron uptake proteins, among which Transferrin Receptor‐1 (TFR‐1). Accumulating evidence has proven that overexpression of TFR‐1 and high Iron concentration, are both widespread condition of cancer cells, both essential to tumour onset and progression. We studied TFR‐1 and PCNA immunohistochemical expression in fifteen (15) Canine osteoblastic osteosarcomas (COS). After immunohistochemical staining, counting of TFR‐1 positive cells by two independent observers showed that 85%–95% of neoplastic cells were strongly labelled at cytoplasmic level by anti‐TFR‐1 antibody in all examined COS. Furthermore, 70%–80% of neoplastic cells were positively labelled at the nuclear level by PCNA. Surprisingly, about 100% of intratumour vascular endothelial cells were also positive, whereas extratumour vascular endothelial cells were negative. The latter is an interesting finding, as TFR‐1 is usually not expressed in normal vasculature, with the exception of normal brain vascular endothelium, where it allows transport of transferrin, and thus iron, into tissues, suggesting a similar function here to support cancer growth. The early results presented highlight the relevance of TFR‐1 expression in canine OS, suggesting therapies involving both TFR‐1 and Iron metabolisms in dogs with osteosarcoma should be developed., Neoplastic cells have higher requirements of iron than normal cells. For that reason, neoplastic cells over‐express genes encoding for iron uptake proteins, including Transferrin Receptor‐1 (TFR‐1). We studied TFR‐1 immunohistochemical expression in ten Canine osteoblastic osteosarcomas (OS). In all examined OS, 85–95% of neoplastic cells showed a strong cytoplasmic immunostaining with anti‐TFR‐1 antibody. Surprisingly, about 100% of intratumor vascular endothelial cells were also positive, whilst extra‐tumor vascular endothelial cells were negative. The early results presented highlight the relevance of TFR‐1 expression in canine OS, suggesting therapies involving both TFR‐1 and Iron metabolisms in dogs with osteosarcoma should be developed.

La sicurezza alimentare della filiera produttiva del miele è tutelata da procedure di controllo stabilite a livello comunitario, che verificano la conformità dell’alimento rispetto ai requisiti previsti in tema di tutela della salute pubblica e utilizzano metodi e tecniche di monitoraggio, sorveglianza, verifica, audit ispezione, campionamento e analisi del prodotto. Inoltre, la consultazione del Piano Nazionale Integrato quadriennale approvato in Italia dalla Conferenza Stato-Regioni,fornisce informazioni sul sistema dei controlli ufficiali attuati sul prodotto. Il Piano Nazionale Residui (PNR) del Ministero della Salute è attuato in via permanente per rilevare nel miele residui di sostanze farmacologicamente attive e contaminanti.Inoltre dal 2005 l’UE ha attuato nei Paesi membri un “Piano di Controllo coordinato sulle pratiche fraudolente nella commercializzazione di prodotti ittici e del miele”. Nel caso del prodotto biologico, oltre alle norme generali di produzione agricola alla produzione biologica animale, si applicano norme di produzione animale riguardo all’origine delle api e alle pratiche apistiche. Infine il Reg. UE n. 1169/11, relativo alle informazioni da riportare in etichetta per il consumatore,prescrive che sul prodotto confezionato, oltre alle indicazioni obbligatorie, il produttore possa inserire informazioni facoltative, come la data di produzione o le indicazioni per la conservazione.