Epithelial Cells Of Urinary Bladder Research Articles

Coinfection with canine distemper virus and canine circovirus in a dog in Brazil.

Canine distemper virus (CDV) and canine circovirus (CanineCV) have been described worldwide in multi-systemic disease in dogs. Both agents may be occasionally associated with other viral pathogens, but reports of coinfection by CDV and CanineCV associated with disease are rare. In this article, we report a coinfection between CDV and CanineCV detected during an investigation of viral agents involved in multisystemic disease in dogs in Southern Brazil. Molecular testing by PCR in lungs and intestines of 77 dogs necropsied in pathology services (2015-2020) revealed several single and mixed viral infections, including a CDV/CanineCV coinfection. In the case reported here, gross and histological findings were compatible with CDV pathology (bronchointerstitial pneumonia and viral intracytoplasmatic inclusions in pneumocytes and transitional epithelial cells of urinary bladder). CanineCV DNA and CDV antigens were detected in lung and intestine fragments by PCR and immunohistochemistry, respectively. CanineCV PCR amplicons subjected to nucleotide sequencing showed > 98.6% nucleotide identity with CanineCV sequences from GenBank. Although the role of CanineCV in the pathogenesis of the reported case could not be determined, our results show that CanineCV may be associated with other viral infections in cases of multisystemic disease in dogs. These results reinforce the circulation of CanineCV in dogs in Brazil and highlight the importance of including this virus in the list of differential diagnoses of respiratory and gastroenteric infectious diseases in dogs.

CHARACTERISTICS OF PILI HEMAGLUTININ PROTEIN AND ITS ROLE IN THE PATHOGENESIS OF URINARY TRACT INFECTION WITH UROPATHOGENIC ESCHERICHIA COLI

Urinary tract infection (UTI) is one of the most common infectious diseasesencountered in the community. The bacteria most frequently implicated as the causes ofUTI are the Gram-negative bacteria, especially Escherechia coli. Early phase of thepathogenesis of the infection constitutes adhesion of bacteria onto the epithelial cells ofurinary tract. SDS-PAGE examination was carried out to investigate molecule ofhemagglutinin protein and hemagglutination (HA) test continued by adhesion andinhibited adhesion tests.The observation of molecular weight of pili E. coli protein molecule byexplorative examination using SDS-PAGE showed on E. coli strip that the mostprominent molecular weights (MW) of the proteins were 61 kDa, 37 kDa, 30 kDa, and 20kDa. Purification by electro-elusion was done to proteins of the MW of 20 kDa, 37 kDaand 61 kDa. The HA test results indicated the 61 kDa, 37 kDa, and 20 kDa werehemagglutinin proteins and one which subsequently applied as sub unit proteins ofUropathogenic E. coli (UPEC) was the 37 kDa. Adhesion test of UPEC bacteria carriedout on urinary bladder epithelial cells of rabbit showed a significant correlation betweenadhesion index and various doses of subunit proteins of UPEC 37 kDa as coat of urinarybladder epithelial cells. Spearman rank correlation test and regression/non regressionanalysis confirmed an exponential decrease of UPEC adhesion to urinary tract epithelialcells as a response to the increasing doses of coating protein. This finding points out thatpili hemagglutinin protein subunit of UPEC 37 kDa was an adhesive molecule. Pilihemagglutinin protein subunit of UPEC 37 kDa is an adhesive molecule, which plays arole in adherence of UPEC to epithelial cells of urinary bladder at the early pathogenesisof urinary tract infection.

Extracellular Ca2+ regulates the stimulus-elicited ATP release from urothelium

Accumulating evidence shows that the epithelial cells in urinary bladder (urothelium) serve as a sensory organ in micturition and/or in nociception pathway by releasing ATP in response to mechanical and/or chemical stimuli. Here, we compared the effects of capsaicin, acetylcholine, and prostaglandin E2 receptor EP1 agonist (ONO-DI-004) on the urothelial ATP release in primary cultured mouse urothelial cells in low Ca2+ medium. All of these chemicals induced a gradual ATP release from urothelium, implying that the downstream Ca2+ release from endoplasmic reticulum could trigger the ATP release. Consistent with this suggestion, blockade of inositol 1,4,5-triphosphate receptor reduced the distention-induced ATP release from urothelial tissues. The distention-induced ATP release was not affected by tetrodotoxin. However, an increase in extracellular Ca2+ diminished both chemical- and distention-induced ATP release from urothelium. Thus raising the extracellular Ca2+ concentration was found to inhibit stimulation-evoked ATP urothelial release.

Effect of osmotic stress on expression of a putative facilitative urea transporter in the kidney and urinary bladder of the marine toad,Bufo marinus

Anuran amphibians accumulate a large amount of urea in their extracellular fluids to avoid a severe dehydration under dry and hyper-saline environments. To clarify the mechanisms of urea retention, we examined structure and distribution of the urea transporter (UT) in the kidney of the marine toad (Bufo marinus), and its expression in the kidney and urinary bladder following exposure to dry and hyper-saline conditions by means of cDNA cloning, semi-quantitative RT-PCR, immunoblot analysis and immunohistochemistry. The Bufo UT cDNA cloned from the kidney encodes a 390-amino-acid residue protein, which is 80% identical to Rana esculenta UT with the functional characteristics of a urea transporter. The Bufo UT mRNA was abundantly expressed in the kidney and urinary bladder, but not in the skin. In immunoblot analysis using a specific antibody raised against the Bufo UT, a 52 kDa protein similar to the glycosylated forms of mammalian UT-A2 ( approximately 55 kDa) was detected in extracts from plasma membrane fractions of the kidney and urinary bladder. When toads were acclimated to dry and hyper-saline environments for 7 days, UT mRNA expression was upregulated in the kidney and urinary bladder and there was an elevated plasma urea concentration and osmolality. Immunohistochemistry showed that the UT was specifically localized on the apical membrane of the early distal tubule, known to be the diluting segment, in the kidney and the epithelial cells of urinary bladder. Immunoreactive cells were not detected along the late distal tubule, the connecting tubule or the collecting duct in the kidney. The present findings suggest that the Bufo UT probably contributes to urea transport in the kidney and urinary bladder in response to hyperosmotic stresses such as body fluid hypertonicity and dehydration.

Viruses and interstitial cystitis: adenovirus genomes cannot be demonstrated in urinary bladder biopsies.

Microbes may be involved in the pathogenesis of interstitial cystitis (IC). Adenoviruses and BK virus (BKV) can infect epithelial cells in urinary bladder and they are causative agents for hemorrhagic cystitis. We therefore studied the presence of adenovirus and BKV genomes in urinary bladder tissue specimens of patients with IC using polymerase chain reaction (PCR) and in situ hybridization (ISH). Controls were specimens from cases with transitional cell carcinoma of the bladder. Nucleic acids were extracted from paraffin sections of the bladder tissue for PCR. Primers detecting all adenovirus types were used. In situ hybridization was carried out for the paraffin sections using digoxigenin-labeled DNA probes for adenovirus and BKV. The adenovirus DNA PCR was able to detect one to two infected cells/specimen. All the seven IC cases studied and six controls were negative for adenovirus DNA by PCR and ISH. The ISH test for BKV genomes was also considered negative in IC cases and controls. The specimens which were negative in PCR tests yielded a signal with beta-globin primers, thus being amplifiable. We conclude that adenovirus and BKV do not play a major pathogenetic role in interstitial cystitis.

The Role of Cells Versus Matrix in Bone Induction

Earlier observations indicated that epithelial cells of urinary bladder, and transformed epithelial cells from human amnion (FL), epidermal carcinoma (HeLa), etc. can induce ectopic endochondral bone formation when implanted into the skeletal muscle of immunosuppressed or autologous animals. Such epithelial cells are associated with little matrix. Bone-inducing activity was also demonstrated in cultured osteosarcoma cells of murine and human origin or extracts thereof, and it is notable that these bone-inducing osteosarcoma cells grow in vitro with little matrix production. Finally, electron microscopy of in vitro cartilage induction showed that decalcified rodent bone that had been extensively extracted to remove cells still contained devitalized cells and cell fragments some of which made contact with inducible, cartilage-forming mesenchymal cells that had migrated in from cocultured muscle. Suggestions: These observations suggest that: 1) the bone-inducing agent(s) of both epithelial and mesenchymal cells may reside mostly in cells rather than matrix. Thus it may be premature to assume that bone "matrix" is the major source of bone-inducing agent in decalcified bone until the osteoinductive activity of residual bone cells has been assessed; and 2) that the osteoinductive agent, whether residing in epithelial cells or bone cells, may be the same or a similar factor operating through the same mechanism.