Several immunohistochemical markers have been used to define the differentiation pattern of urothelial cell tumors of the urinary bladder. We investigated the expression of the recently characterized uroplakin (UP) IIIb, an urothelium-specific and differentiation-dependent protein, in 39 urothelial tumors of the urinary bladder in cows that had suffered from chronic enzootic hematuria for several years. Bovine papillomavirus type 2 DNA was amplified and UP IIIb protein was detected in all these tumors. In papillomas and papillary carcinomas, UP IIIb expression was mostly seen as superficial staining; luminal and peripheral patterns were also observed. In nonpapillary carcinomas, UP IIIb appeared to define clearly the cell membrane lining intercellular and intracellular lumina as well as the cell borders in deeper cell layers. In benign and malignant lesions, an intracytoplasmic immunoreactivity was also detected. Coarse intracytoplasmic UP IIIb-positive material close to the nucleus occurred in some malignant cells. Focally strong membraneous immunostaining that marked single cells with complete ringlike peripheral pattern was seen. Although UP IIIb expression does not seem to correlate with the biological behavior of urothelial tumors, it appears to be a highly sensitive marker for bovine urothelial tumors.
"e l s e v i e r . c o m / l o c a t e / f o o d c h e m lesions of animals grazing on bracken fern have been reported (Carvalho, Pinto, & Peleteiro, 2006; Roperto et al., 2005, 2010; Sardon et al., 2005). Because of the increased risk for oesophageal and gastric cancer in humans living in areas rich in bracken fern and exposed to foods contaminated by Pta, the need to avoid the distribution of Pta into the food chain has become a crucial topic in the food safety investigations (Alonso-Amelot & Avendano, 2002; Shahin, Smith, & Prakash, 1999). "
[Show abstract][Hide abstract] ABSTRACT: We describe a new method to detect trace levels of ptaquiloside (Pta), a major carcinogen of bracken fern in biological samples such as milk from farm animals. The method involves the absorption of analyte on carbograph followed by elution with solvents mixtures. The unstable analyte is then converted into Br-Pt (II), which is specific for Pta, as it is not a natural decay product of the glycoside in aqueous media. An internal standard, the Br-pterosine-d2, prepared in our laboratories has been used. Detection and quantification are possible with gas chromatography/mass spectrometry (GC/MS) in single ion monitoring mode (SIM). The detectable amount is in the range of ppb. The method allowed us to detect Pta not only in the milk from bracken fern-poisoned cattle but also, for the first time, in the milk from healthy farm animals such as sheep, goat, horse, and donkey mares.
"Recently, it has been shown that the autophagic pathway is involved in trafficking events that activate innate and adaptive immune responses to viruses, bacteria and parasites (Levine and Deretic, 2007; Schmid and Mu¨nz, 2007; Xu et al., 2008). Cattle with urinary bladder tumours may prove useful as animal models for the study of biochemical markers in oncogenesis (Roperto et al., 2005, 2007). "
[Show abstract][Hide abstract] ABSTRACT: The expression of sigma-2 receptors was investigated in nine urothelial tumours of the urinary bladder of cattle. Each tumour was associated with the presence of DNA of bovine papillomavirus type-2 (BPV-2) and expression of the E5 viral oncoprotein. Five tumours were classified as low-grade carcinoma on the basis of morphological criteria and calculation of mean nuclear area (MNA) and mean nuclear perimeter (MNP). Four tumours were classified as high-grade carcinoma. Sigma-2 receptors were overexpressed in both types of carcinoma. In control normal bovine bladder tissue the density of receptors (expressed as the B(max)) was 0.37 pmol/mg of protein. Low-grade carcinomas had a mean B(max) of 1.37+/-0.32 pmol/mg of protein (range 1.03-1.86) and in high-grade carcinomas the mean B(max) was 10.9+/-2.8 pmol/mg of protein (range 8.2-14). The difference in B(max) between low- and high-grade carcinomas was statistically significant (P=0.0001).
"Ten samples of neoplastic urothelium were collected at public slaughterhouses from cows aged 4e24 years that had suffered from chronic enzootic haematuria. All animals were raised as mountain cattle in the south of Italy (Roperto et al., 2005) and were known to have grazed pastures rich in bracken fern. Normal (control) bladder mucosa was obtained from five healthy cows aged 4e14 years that had grazed lowland pastures without bracken. "
[Show abstract][Hide abstract] ABSTRACT: The up-regulation of ferritin heavy chain (FHC) is reported in six papillary and in four invasive urothelial tumours of the urinary bladder of cattle grazing on mountain pastures rich in bracken fern. All tumours contained sequence of bovine papillomavirus type-2 (BPV-2) as determined by polymerase chain reaction (PCR) analyses and validated by direct sequencing of the amplified products. The oncoprotein E5 was also detected in these tumours by immunoprecipitation and by immunofluorescence and laser scanning confocal microscopy. Expression of FHC was evaluated by western blot analysis, reverse transcriptase (RT) PCR, real-time RT-PCR and immunohistochemistry. The oligonucleotide sequence of the bovine ferritin amplicons was identical to that of human ferritin. Nuclear overexpression of p65, an important component of nuclear factor kappaB (NF-kappaB) transcription factors, was also observed. These findings suggest that FHC up-regulation may be mediated by activation of NF-kappaB and that in turn this may be related to the resistance of bovine papillomavirus type-2 (BPV-2) infected urothelial cells to apoptosis.
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