GRP78 expression in canine mammary tumors: association with malignancy

78-kDa glucose-regulated protein (GRP78) is over-expressed in human breast carcinomas. GRP78 expression was studied in 40 spontaneous canine mammary tumors and evaluated in relation to tumor histological type, mode of growth, grade, lymph node metastases and distant metastases. All tumors exhibited GRP78 immunostaining. In the normal canine mammary gland, GRP78 was also expressed although not in all cases. In carcinomas GRP78 was detected in the cytoplasm in more than 50% of tumor cells in the vast majority of cases (87.5%). There was a significant association between the absence of squamous differentiation (P = 0.02) and GRP78 over-expression, but no association with other clinico-pathological features. GRP78 was often co-expressed with galectin-3 in canine mammary tumors (CMT).

Expression of E-cadherin, P-cadherin and beta-catenin in canine malignant mammary tumours in relation to clinicopathological parameters, proliferation and survival

Cadherin-catenin complexes play a critical role in intercellular adhesion, and their altered expression has been implicated in tumour progression. In this study, the expression of E-cadherin, P-cadherin and beta-catenin was analysed in 65 canine malignant mammary tumours and correlated with clinicopathological parameters, proliferation and survival. Reduction in E-cadherin expression was significantly associated with increased tumour size, high histological and invasion grades, lymph node metastasis and high mitotic index. Reduced beta-catenin expression was associated with high histological and invasion grades. Anomalous expression of P-cadherin was only associated with invasion. In 39 cases for which follow-up data were available, reduced E-cadherin and beta-catenin expression was significantly associated with shorter overall survival and disease free survival. Abnormal expression of adhesion molecules is a common phenomenon in canine mammary malignant tumours and may play a central role in tumour progression.This work was supported by the Centro de Ciência Animal e Veterinária (CECAV) – University of Trás os Montes e Alto Douro (UTAD), Vila Real, Portugal and by Portuguese Science and Technology Foundation, Project POCTI/CVT/57795/2004

Presence and significance of Helicobacter spp. in the gastric mucosa of Portuguese dogs

Background: Non-Helicobacter pylori Helicobacters (NHPH) are also able to cause disease in humans. Dogs are a natural reservoir for many of these species. Close and intense human contact with animals has been identified as a risk factor and therefore, an important zoonotic significance has been attributed to NHPH. Methods: To determine the prevalence of Helicobacter species and the gastric histopathological changes associated, gastric mucosa samples of 69 dogs were evaluated. Results: Only one dog presented a normal histopathological mucosa with absence of spiral-shaped organisms. A normal gastric mucosa and the presence of spiral-shaped bacteria was observed in two dogs. All remaining animals presented histopathological changes representative of gastritis. Helicobacter species were detected in 60 dogs (87.0%) by at least one detection method. Histological, histochemical and immunohistochemical evaluations revealed that Helicobacter spp. were present in 45 (65.2%), 52 (75.4%) and 57 (82.6%) dogs, respectively. Spiral-shaped bacteria were detected by qPCR analysis in 33 (47.8%) dogs. H. heilmannii-like organisms were identified in 22 animals (66.7%) and predominantly in the antral gastric region. H. salomonis was the second most prevalent species (51.5%) although it was mainly found in association with other Helicobacter spp. and in the body gastric region. H. bizzozeronii and H. felis were less frequently detected. Conclusions: It was concluded that, despite the high incidence and worldwide distribution of gastric NHPH in dogs, the presence of specific Helicobacter species may vary between geographic regions. NHPH infections were significantly accompanied by mild to moderate intraepithelial lymphocyte infiltration and mild to moderate gastric epithelial injury, but a clear relationship between gastritis and Helicobacter infection could not be established

Trace metals and over-expression of metallothioneins in bladder tumoral lesions: a case-control study

<p>Abstract</p> <p>Background</p> <p>Previous studies have provided some evidence of a possible association between cancer and metallothioneins. Whether this relates to an exposure to carcinogenic metals remains unclear.</p> <p>Methods</p> <p>In order to examine the association between the expression of metallothioneins and bladder tumors, and to compare the levels of arsenic, cadmium, chromium, lead and nickel in animals with bladder tumors and animals without bladder tumors, 37 cases of bovine bladder tumors and 17 controls were collected. The detection and quantification of metallothioneins in bladder tissue of both cases and controls was performed by immunohistochemistry. And the quantification of metals in tissue and hair was assessed by inductively coupled plasma – mass spectrometry.</p> <p>Results</p> <p>Increased expression of metallothioneins was associated with bladder tumors when compared with non-tumoral bladder tissue (OR = 9.3, 95% CI: 1.0 – 480). The concentrations of cadmium, chromium, lead and nickel in hair of cases were significantly higher than those of controls. However, as for the concentration of metals in bladder tissue, the differences were not significant.</p> <p>Conclusion</p> <p>Though the sample size was small, the present study shows an association between bladder tumors and metallothioneins. Moreover, it shows that concentrations of metals such as cadmium, chromium, lead and nickel in hair may be used as a biomarker of exposure.</p

CD117 immunoexpression in canine mast cell tumours: correlations with pathological variables and proliferation markers

<p>Abstract</p> <p>Background</p> <p>Cutaneous mast cell tumours are one of the most common neoplasms in dogs and show a highly variable biologic behaviour. Several prognosis tools have been proposed for canine mast cell tumours, including histological grading and cell proliferation markers. CD117 is a receptor tyrosine kinase thought to play a key role in human and canine mast cell neoplasms. Normal (membrane-associated) and aberrant (cytoplasmic, focal or diffuse) CD117 immunoexpression patterns have been identified in canine mast cell tumours. Cytoplasmic CD117 expression has been found to correlate with higher histological grade and with a worsened post-surgical prognosis. This study addresses the role of CD117 in canine mast cell tumours by studying the correlations between CD117 immunoexpression patterns, two proliferation markers (Ki67 and AgNORs) histological grade, and several other pathological variables.</p> <p>Results</p> <p>Highly significant (p < 0,001) correlations were found between CD117 immunostaining patterns and histological grade, cell proliferation markers (Ki67, AgNORs) and tumoral necrosis. Highly significant (p < 0,001) correlations were also established between the two cellular proliferation markers and histological grade, tumour necrosis and epidermal ulceration. A significant correlation (p = 0.035) was observed between CD117 expression patterns and epidermal ulceration. No differences were observed between focal and diffuse cytoplasmic CD117 staining patterns concerning any of the variables studied.</p> <p>Conclusion</p> <p>These findings highlight the key role of CD117 in the biopathology of canine MCTs and confirm the relationship between aberrant CD117 expression and increased cell proliferation and higher histological grade. Further studies are needed to unravel the cellular mechanisms underlying focal and diffuse cytoplasmic CD117 staining patterns, and their respective biopathologic relevance.</p

Infection with carcinogenic helminth parasites and its production of metabolites induces the formation of DNA-adducts

Background: Infections classified as group 1 biological carcinogens include the helminthiases caused by Schistosoma haematobium and Opisthorchis viverrini. The molecular mediators underlying the infection with these parasites and cancer remain unclear. Although carcinogenesis is a multistep process, we have postulated that these parasites release metabolites including oxysterols and estrogen-like metabolites that interact with host cell DNA. How and why the parasite produce/excrete these metabolites remain unclear. A gene encoding a CYP enzyme was identified in schistosomes and opisthorchiids. Therefore, it is reasonable hypothesized that CYP 450 might play a role in generation of pro-inflammatory and potentially carcinogenic compounds produced by helminth parasites such as oxysterols and catechol estrogens. Here, we performed enzymatic assays using several isoforms of CYP 450 as CYP1A1, 2E1 and 3A4 which are involved in the metabolism of chemical carcinogens that have been associated with several cancer. The main aim was the analysis of the role of these enzymes in production of helminth-associated metabolites and DNA-adducts. Method: The effect of cytochrome P450 enzymes CYP 1A1, 2E1 and 3A4 during the interaction between DNA, glycocholic acid and taurochenodeoxycholate sodium on the formation of DNA-adducts and metabolites associated with urogenital schistosomiasis (UGS) and opisthorchiasis was investigated in vitro. Liquid chromatography/mass spectrometry was used to detect and identify metabolites. Main findings: Through the enzymatic assays we provide a deeper understanding of how metabolites derived from helminths are formed and the influence of CYP 450. The assays using compounds similar to those previously observed in helminths as glycocholic acid and taurochenodeoxycholate sodium, allowed the detection of metabolites in their oxidized form and their with DNA. Remarkably, these metabolites were previously associated with schistosomiaisis and opisthorchiasis. Thus, in the future, it may be possible to synthesize this type of metabolites through this methodology and use them in cell lines to clarify the carcinogenesis process associated with these diseases. Principal conclusions: Metabolites similar to those detected in helminths are able to interact with DNA in vitro leading to the formation of DNA adducts. These evidences supported the previous postulate that imply helminth-like metabolites as initiators of helminthiases-associated carcinogenesis. Nonetheless, studies including these kinds of metabolites and cell lines in order to evaluate its potential carcinogenic are required.This work was financed by FEDER - Fundo Europeu de Desenvolvimento Regional funds through the COMPETE 2020 - Operacional Programme for Competitiveness and Internationalisation (POCI), Portugal 2020, and by Portuguese funds through FCT - Fundação para a Ciência e a Tecnologia, in the framework of the project, Institute for Research and Innovation in Health Sciences” (POCI-01-0145-FEDER-007274). The FCT and FEDER (European Union) also supported these studies through project number IF/00092/2014/CP1255/CT0004info:eu-repo/semantics/publishedVersio

TWIST1 Gene: First Insights in Felis catus

TWIST1 is thought to be a novel oncogene. Understanding the molecular mechanisms regulating the TWIST1 gene expression profiles in tumor cells may give new insights regarding prognostic factors and novel therapeutic targets in veterinary oncology. In the present study we partially isolated the TWIST1 gene in Felis catus and performed comparative studies. Several primer combinations were used based on the alignments of homologous DNA sequences. After PCR amplification, three bands were obtained, purified and sequenced. Several bioinformatic tools were utilized to carry out the comparative studies. Higher similarity was found between the isolated TWIST1 gene in Felis catus and Homo sapiens (86%) than between Homo sapiens and Rattus norvegicus or Mus musculus (75%). Partial amino acid sequence showed no change in the four species analyzed. This confirmed that coding sequences presented high similarity (~96%) between man and cat. These results give the first insights regarding the TWIST1 gene in cat but further studies are required in order to establish, or not, its role in tumor formation and progression in veterinary oncology

In vivo biocompatibility and biodegradability of dextrin-based hydrogels

The in vivo biocompatibility of dextrin hydrogels obtained by polymerization of dextrin-hydroxyethylmethacrylate (dextrin-HEMA) and dextrin-vinyl acrylate (dextrin-VA) are reported in this work. The histological analysis of subcutaneous implants of these hydrogels, featuring inflammatory and reabsorption events, were carried out over a 16-week period in mice. The dextrin-HEMA hydrogel was quickly and completely degraded and reabsorbed, whereas the dextrin-VA degradation occurred slowly and a thin fibrous capsule surrounded the nondegradable hydrogel. The dextrin-HEMA was degraded after 16 weeks with only mild inflammation and a few detectable foamy macrophages around the implant. These events were followed by complete resorption and no sign of capsule formation or fibrosis associated to the implants. The results indicate that the dextrin hydrogels are biocompatible because no toxicity on the tissues surrounding the implants was found. It may be speculated that a controlled degradation rate of the hydrogels may be obtained by grafting dextrin to HEMA and VA in different proportions.Funding from FCT through POCTI program is acknowledged. The authors Susana Moreira and Rui M. Gil da Costa are recipients of a PhD fellowship from Fundacao para a Ciencia e a Tecnologia (FCT, Portugal)

Sialyl Lewis x expression in canine malignant mammary tumours: correlation with clinicopathological features and E-Cadherin expression

<p>Abstract</p> <p>Background</p> <p>Sialyl Lewis x (sLe^x) antigen is a carbohydrate antigen that is considered not only a marker for cancer but also implicated functionally in the malignant behaviour of cancer cells. Overexpression of sLe^xis associated with enhanced progression and metastases of many types of cancer including those of the mammary gland. Canine mammary tumours can invade and give rise to metastases via either lymphatic or blood vessels.</p> <p>E-Cadherin is specifically involved in epithelial cell-to-cell adhesion. In cancer, E-Cadherin underexpression is one of the alterations that characterizes the invasive phenotype and is considered an invasion/tumour suppressor gene. Partial or complete loss of E-Cadherin expression correlates with poor prognosis in canine malignant mammary cancer.</p> <p>The aim of this study was to analyse the sLe^xexpression in canine malignant mammary tumours and to evaluate if the presence of sLe^xcorrelates with the expression of E-Cadherin and with clinicopathological features.</p> <p>Methods</p> <p>Fifty-three cases of canine mammary carcinomas were analysed immunohistochemically using monoclonal antibodies against sLe^x(IgM) and E-Cadherin (IgG). The clinicopathological data were then assessed to determine whether there was a correlation with sLe^xtumour expression. Double labelled immunofluorescence staining was performed to analyse the combined expression of sLe^xand E-Cadherin.</p> <p>Results</p> <p>sLe^xexpression was consistently demonstrated in all cases of canine mammary carcinomas with different levels of expression. We found a significant relationship between the levels of sLe^xexpression and the presence of lymph node metastases. We also demonstrated that when E-Cadherin expression was increased sLe^xwas reduced and vice-versa. The combined analysis of both adhesion molecules revealed an inverse relationship.</p> <p>Conclusion</p> <p>In the present study we demonstrate the importance of sLe^xin the malignant phenotype of canine malignant mammary tumours. Our results support the use of sLe^xas a prognostic tumour marker in canine mammary carcinomas. Furthermore, we showed that sLe^xand E-Cadherin expression were inversely correlated. Future studies are warranted to clarify the molecular mechanism underlying the relation between sLe^xand E-Cadherin in canine mammary carcinoma cells which represents an important comparative model to woman breast cancer.</p