OBJECTIVE: The methodology of cell blocks (CBs) has long been an integrated part of cytology. However, there are very few data on CBs derived from urine. Their main disadvantage is a lack of cellularity, which limits their broader clinical applicability. Factors affecting cellular adequacy in urine remain unclear. We assessed the impact of basic clinical and cytopathological factors on the adequacy of cellularity in urinary CBs. METHODS: Freshly voided urine was collected from 401 consecutive individuals. Of these, 167 patients were diagnosed with urothelial carcinoma. The remaining 234 patients had various benign urological conditions. Papanicolaou classes were determined and CBs produced. Cellular adequacy was assigned to each CB (acellular, hypocellular, moderate cellularity, high cellularity), and moderately and highly cellular CBs were considered as adequate. Several factors were analysed to find any correlation with the adequacy of the cellularity. RESULTS: In univariate analysis, seven factors significantly correlated with the adequacy of the CBs. In the multivariate model, positive sediment (OR = 3.7), female sex (OR = 2.7), positive urinary cytology (OR = 2.6) and positive leucocyturia (OR = 2.1) were independent predictors of adequate cellularity. Positive predictive value and negative predictive value of the model were 65.0% and 77.7%, respectively. CONCLUSIONS: We determined four clinical and cytopathological factors which independently predict adequate cellularity in urinary CBs. Based on these results, several clinical situations have been proposed, in which the highest probability of adequate cellularity in urinary CBs can be achieved.

• Klíčová slova
• adequate cellularity, immunocytochemistry, urinary bladder, urinary cell block, urinary cytology, urothelial carcinoma,
• MeSH
• cytodiagnostika metody   MeSH
• karcinom z přechodných buněk diagnóza   patologie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• moč cytologie   MeSH
• močový měchýř patologie   MeSH
• nádory močového měchýře diagnóza   patologie   MeSH
• následné studie MeSH
• senioři MeSH
• urologické nádory diagnóza   patologie   MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH

OBJECTIVE: Immunocytochemistry has attained a marginal role in urology so far. Combining the morphological and immunophenotypical changes of the urothelial cells retrieved from urine is a logical approach. The study aimed to analyse the diagnostic potential of immunocytological staining in the detection of high-grade and low-grade urothelial carcinoma. METHODS: Freshly voided urine was collected from 152 consecutive individuals, cytology classes were determined and cell blocks produced. A total of 77 patients were diagnosed with urothelial carcinoma and 75 patients had various benign urological conditions. Immunocytochemistry was performed using four antibodies: p53, MCM2, MCM5 and Ki-67. A diagnostic power to detect low grade and high-grade urothelial carcinoma was analysed for each antibody and their combinations with cytology. RESULTS: There were no significant differences between patients with low-grade tumours and control group. Antibodies p53 and Ki-67 slightly improved the sensitivity of urinary cytology while maintaining its specificity. The best negative predictive value was demonstrated in combinations of cytology and MCM5 (88.9%) and cytology, p53 and MCM5 (90.6%). In the diagnosis of high-grade tumours, all antibodies apart from MCM2 yielded better sensitivity and specificity than cytology alone (receiver operating characteristic curves: p53 = 0.853, MCM5 = 0.931, and Ki-67 = 0.895). Combined with cytology, the sensitivities went even higher for the cost of lower specificity. The best diagnostic performance was observed in the combination of MCM5 and Ki-67 (sensitivity = 96.2%; specificity = 80%). CONCLUSIONS: Immunocytochemistry with p53, MCM5 and Ki-67 antibodies can improve the diagnostic power of urinary cytology in the detection and follow-up of urinary bladder urothelial carcinoma.

• Klíčová slova
• diagnosis, follow-up, immunocytochemistry, urinary bladder, urinary cell block, urinary cytology, urothelial carcinoma,
• MeSH
• antigen Ki-67 imunologie   MeSH
• cytodiagnostika * MeSH
• imunohistochemie MeSH
• lidé MeSH
• MCM komplex, komponenta 2 imunologie   MeSH
• moč cytologie   MeSH
• nádorové biomarkery metabolismus   MeSH
• nádorový supresorový protein p53 imunologie   MeSH
• nádory močového měchýře diagnóza   patologie   MeSH
• proteiny buněčného cyklu imunologie   MeSH
• protilátky nádorové imunologie   MeSH
• ROC křivka MeSH
• senioři MeSH
• senzitivita a specificita MeSH
• stupeň nádoru MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• antigen Ki-67 MeSH
• MCM komplex, komponenta 2 MeSH
• MCM2 protein, human MeSH Prohlížeč
• MCM5 protein, human MeSH Prohlížeč
• nádorové biomarkery MeSH
• nádorový supresorový protein p53 MeSH
• proteiny buněčného cyklu MeSH
• protilátky nádorové MeSH

Currently, we have assessed the neuronal control of the urinary bladder in radiation cystitis and whether interstitial cells contribute to the condition. Fourteen days after bladder irradiation (20 Gy), rats were sedated and the urinary bladder was cut into two sagittal halves where electrical field stimulation (EFS; 5-20 Hz) was applied on the pelvic nerve afferents or stretch (80 mN) on one-half of the bladder, while contractions were registered on the contralateral half of the bladder in the absence and presence of increasing doses of imatinib (1-10 mg/kg; inhibitor of c-kit-positive interstitial cells), atropine (1 mg/kg; to block muscarinic M3 receptors), or pyridoxalphosphate-6-azophenyl-2',4'-disulfonic acid (2 mg/kg; P2X1 purinoceptor antagonist). Urinary bladders were also excised for organ bath experiments, Western blot, quantitative polymerase chain reaction, and immunohistochemistry. In vivo, EFS contractions were enhanced after irradiation, and imatinib (1-10 mg/mg) inhibited contractions by EFS and stretched dose-dependently in controls but not in irradiated bladders. In the irradiated bladder in vitro, atropine resistance was increased and imatinib (100 µM) inhibited contractions by EFS and agonists (ATP, methacholine) in irradiated bladders and controls. The urinary bladder expressions of P2X1 purinoceptors, muscarinic M3 receptor, choline acetyltransferase, c-kit, and the agonist of c-kit, stem cell factor, were not changed by irradiation. In conclusion, bladder irradiation affects several levels of neuronal control of the urinary bladder. Interstitial cells may contribute to some of the symptoms associated with radiation cystitis.

• MeSH
• cystitida patologie   patofyziologie   MeSH
• krysa rodu Rattus MeSH
• močový měchýř patologie   patofyziologie   MeSH
• modely nemocí na zvířatech MeSH
• neurony patologie   MeSH
• svalová kontrakce MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Peritoneal exudate cells (PEC) from conventional donors displayed in vitro the cytotocix effects on the rat sarcoma cell lines LWI3K2, LW13K3, LW13K4 and RSL but they were not cytotoxic to fibroblasts derived from normal rats of the same strains. PEC were also cytotoxic to human urinary bladder carcinoma cells (T-24) grown in vitro. Cytotoxicity was detectable already at the effector to target cell ratio of 1 : 1, and the effector cells from syngeneic, allogeneic and xenogeneic donors were equally effective. The cytotoxic effect was apparently exerted by macrophages because the addition of lymphocytes from PEC donors did not change the target cells. The addition of serum from PEC donors to a mixture of target and effector cells protected the tumour cells from the destruction by macrophages. Cytotoxicity was blocked by serum from all PEC donors tested (syngeneic, allogeneic and xenogeneic with regard to the target cells) but not by foetal calf serum.

• MeSH
• buněčné linie MeSH
• cytotoxické testy imunologické * MeSH
• exsudáty a transsudáty imunologie   MeSH
• fibroblasty imunologie   MeSH
• inbrední kmeny myší imunologie   MeSH
• makrofágy imunologie   MeSH
• myši MeSH
• nádory močového měchýře imunologie   MeSH
• ptačí sarkom imunologie   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Detergents (Triton X-100, dodecyl sulphate, saponin) added in concentrations of 0.1-10 mg/ml to solution at the outer frog skin surface reversibly reduced potential difference (PD) and short-circuit current (Isc); when added at the inner surface. Triton X-100 initially induced a short increase in the PD and Isc followed by a pronounced decrease similarly as after the application of any of the detergents used. When added to either serosal or mucosal surface of frog urinary bladder, the detergents reversibly abolished the reactivity to vasopressin. Triton X-100 blocked foskolin and cAMP-induced effects on membrane water permeability. The results suggest that hydrophobic elements of membrane play a crucial role in the regulation of membrane permeability for ions and water and of the responsiveness of the cells to vasopressin. Differences in sensitivity to detergents could be observed between the apical and the basolateral cell membrane.

• MeSH
• biologický transport MeSH
• detergenty farmakologie   MeSH
• dodecylsíran sodný farmakologie   MeSH
• draslík farmakokinetika   MeSH
• draslíkové kanály účinky léků   MeSH
• kůže metabolismus   MeSH
• močový měchýř účinky léků   metabolismus   MeSH
• oktoxynol MeSH
• osmóza účinky léků   MeSH
• permeabilita buněčné membrány MeSH
• polyethylenglykoly farmakologie   MeSH
• povrchově aktivní látky farmakologie   MeSH
• Rana temporaria MeSH
• saponiny farmakologie   MeSH
• sodík farmakokinetika   MeSH
• techniky in vitro MeSH
• voda metabolismus   MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• detergenty MeSH
• dodecyl sulfate MeSH Prohlížeč
• dodecylsíran sodný MeSH
• draslík MeSH
• draslíkové kanály MeSH
• oktoxynol MeSH
• polyethylenglykoly MeSH
• povrchově aktivní látky MeSH
• saponiny MeSH
• sodík MeSH
• voda MeSH

Several lines of evidence suggest that in mice the activation of SMAD2/3 signaling by oocyte secreted factors, together with epidermal growth factor receptor (EGFR) activation, is essential to induce cumulus expansion. Here we show that inhibition of EGFR kinase in follicle stimulating hormone (FSH)-stimulated porcine oocyte-cumulus cell complex (OCCs) strongly decreases hyaluronan (HA) synthesis and its retention in the matrix, as well as progesterone synthesis. Although porcine cumulus cells undergo expansion independently of oocytes, we use biochemical and gene expression analyses to show that they do require activation of SMAD2/3 for optimal stimulation of HA synthesis and proteins involved in the organization of this polymer in the expanded matrix. Furthermore, FSH-induced progesterone synthesis by porcine cumulus cells was increased by blocking SMAD2/3 activation. In conclusion, these results support the hypothesis that an FSH-EGF autocrine loop is active in porcine OCCs, and provide the first evidence that the SMAD2/3 signaling pathway is induced by paracrine/autocrine factors in porcine cumulus cells and is involved in the control of both cumulus expansion and steroidogenesis.

• MeSH
• benzamidy farmakologie   MeSH
• C-reaktivní protein metabolismus   MeSH
• chinazoliny farmakologie   MeSH
• dioxoly farmakologie   MeSH
• epidermální růstový faktor metabolismus   MeSH
• erbB receptory antagonisté a inhibitory   metabolismus   MeSH
• folikuly stimulující hormon metabolismus   MeSH
• glukuronosyltransferasa antagonisté a inhibitory   metabolismus   MeSH
• isochinoliny farmakologie   MeSH
• kumulární buňky metabolismus   MeSH
• kyselina hyaluronová biosyntéza   MeSH
• meióza účinky léků   MeSH
• molekuly buněčné adheze antagonisté a inhibitory   metabolismus   MeSH
• myši MeSH
• oocyty enzymologie   fyziologie   MeSH
• prasata MeSH
• progesteron biosyntéza   MeSH
• protein Smad2 antagonisté a inhibitory   metabolismus   MeSH
• protein Smad3 antagonisté a inhibitory   metabolismus   MeSH
• pyridiny farmakologie   MeSH
• pyrroly farmakologie   MeSH
• sérový amyloidový protein metabolismus   MeSH
• signální transdukce účinky léků   MeSH
• tyrphostiny farmakologie   MeSH
• vývojová regulace genové exprese účinky léků   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• 4-(5-benzo(1,3)dioxol-5-yl-4-pyridin-2-yl-1H-imidazol-2-yl)benzamide MeSH Prohlížeč
• 6,7-dimethyl-2-(2E)-3-(1-methyl-2-phenyl-1H-pyrrolo(2,3-b)pyridin-3-yl-prop-2-enoyl)-1,2,3,4-tetrahydroisoquinoline hydrochloride MeSH Prohlížeč
• benzamidy MeSH
• C-reaktivní protein MeSH
• chinazoliny MeSH
• dioxoly MeSH
• epidermální růstový faktor MeSH
• erbB receptory MeSH
• folikuly stimulující hormon MeSH
• glukuronosyltransferasa MeSH
• isochinoliny MeSH
• kyselina hyaluronová MeSH
• molekuly buněčné adheze MeSH
• progesteron MeSH
• protein Smad2 MeSH
• protein Smad3 MeSH
• PTX3 protein MeSH Prohlížeč
• pyridiny MeSH
• pyrroly MeSH
• RTKI cpd MeSH Prohlížeč
• sérový amyloidový protein MeSH
• tyrphostiny MeSH

How the proliferation of the urothelium is regulated is known to a little degree. E. coli lipopolysaccharide (LPS) activates the innate immune response of the urinary bladder via the Toll-like receptor 4 (TLR4) on the urothelium but induces also urothelial proliferation. We wanted to assess whether muscarinic receptors are involved in the regulation of urothelial proliferation triggered by LPS stimulation. Female Fischer 344 rats were instilled with LPS or saline (control) in the urinary bladder in the absence or presence of muscarinic receptor blockade with atropine and regeneration of the urothelium was assessed 4h and 24h later. In the Fischer 344 bladder, urothelial thinning and urothelial caspase 3 up-regulation occurred at 4h after LPS urinary bladder instillation, which were totally blocked in rats pre-treated with atropine. TLR4 was only expressed in blood vessels in the Fischer 344 bladder, while it was also expressed in umbrella cells in the Sprague-Dawley bladder. Proliferation (Ki67 incorporation) of the human urothelial cell line UROtsa was reduced in the presence of the muscarinic receptor antagonists methoctramine (M2/M4-selective) and pirenzepine (M1/M4-selective), while proliferation instead was enhanced in the presence of atropine. In UROtsa cells exposed to LPS for 24h, 4-DAMP (M3/M1/M5-selective) inhibited instead proliferation. In conclusion, muscarinic receptors regulate urothelial proliferation and LPS may induce urothelial apoptosis via muscarinic receptor-dependent pathways. Our findings also suggest that species differences exist in the expressional pattern of TLR4 in the urothelium.

• MeSH
• apoptóza MeSH
• atropin farmakologie   MeSH
• buněčné linie MeSH
• cholinergní látky farmakologie   MeSH
• diaminy farmakologie   MeSH
• Escherichia coli imunologie   MeSH
• kaspasa 3 metabolismus   MeSH
• krysa rodu Rattus MeSH
• lidé MeSH
• lipopolysacharidy imunologie   MeSH
• pirenzepin farmakologie   MeSH
• potkani inbrední F344 MeSH
• potkani Sprague-Dawley MeSH
• přirozená imunita MeSH
• proliferace buněk MeSH
• receptory muskarinové metabolismus   MeSH
• toll-like receptor 4 metabolismus   MeSH
• urotel patologie   fyziologie   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• lidé MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• atropin MeSH
• cholinergní látky MeSH
• diaminy MeSH
• kaspasa 3 MeSH
• lipopolysacharidy MeSH
• methoctramine MeSH Prohlížeč
• pirenzepin MeSH
• receptory muskarinové MeSH
• toll-like receptor 4 MeSH

Polychlorinated biphenyls (PCBs), a structurally diverse group of environmental pollutants, are effective promoters in two-stage cancer models, which implies that epigenetic mechanisms are involved. Inhibition of gap junctional intercellular communication (GJIC) belongs among critical epigenetic events of tumor promotion. We determined the relative potencies of a series of environmentally relevant PCB congeners to inhibit GJIC in vitro in a rat liver epithelial cell line with pluripotent oval cell characteristics. The nonplanar PCBs were potent inhibitors of GJIC, whereas the coplanar PCBs did not inhibit GJIC. We then compared the effects of the coplanar PCB 126 (3,3',4,4',5-pentachlorobiphenyl) and the noncoplanar PCB 153 (2,2',4,4',5,5'-hexachlorobiphenyl) with effects of two model GJIC inhibitors, a tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA) and epidermal growth factor (EGF). In contrast to TPA or EGF, PCB 153 elicited a long-term downregulation of GJIC (up to 48 h). Using Western blot analysis with phospho-specific antibodies, it was found that PCB 153, and not PCB 126, activated mitogen-activated protein kinases ERK1/2; however in contrast to TPA and EGF, this activation was observed at the time points subsequent to GJIC inhibition. Moreover, blocking of ERK1/2 activation did not prevent the GJIC inhibition induced by PCB 153. Therefore, additional intracellular signaling pathways potentially involved in the downregulation of GJIC by PCBs were screened by using specific chemical probes inhibiting serine/threonine kinases, tyrosine kinases, and phospholipases. The inhibition of diacylglycerol lipase partially blocked and the selective inhibition of Src kinases and phosphatidylcholine-specific phospholipase C (PC-PLC) completely blocked the inhibitory effects of the noncoplanar PCB on GJIC, indicating that PC-PLC or sphingomyelinase and Src might be upstream regulators of noncoplanar PCB-induced inhibition of GJIC.

• MeSH
• buněčné linie MeSH
• epidermální růstový faktor toxicita   MeSH
• epitelové buňky účinky léků   enzymologie   MeSH
• játra cytologie   MeSH
• krysa rodu Rattus MeSH
• mezerový spoj účinky léků   enzymologie   MeSH
• mitogenem aktivované proteinkinasy metabolismus   MeSH
• polychlorované bifenyly toxicita   MeSH
• sfingomyelinfosfodiesterasa metabolismus   MeSH
• signální transdukce účinky léků   MeSH
• skupina kinas odvozených od src-genu metabolismus   MeSH
• tetradekanoylforbolacetát toxicita   MeSH
• western blotting MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, U.S. Gov't, P.H.S. MeSH
• Názvy látek
• 2,4,5,2',4',5'-hexachlorobiphenyl MeSH Prohlížeč
• 3,4,5,3',4'-pentachlorobiphenyl MeSH Prohlížeč
• epidermální růstový faktor MeSH
• mitogenem aktivované proteinkinasy MeSH
• polychlorované bifenyly MeSH
• sfingomyelinfosfodiesterasa MeSH
• skupina kinas odvozených od src-genu MeSH
• tetradekanoylforbolacetát MeSH

AIMS: Pathological evaluation of lymphadenectomy specimens plays a pivotal role in accurate lymph node (LN) staging. Guidelines standardising the gross handling and reporting of pelvic LN dissection (PLND) in prostate (PCa) and bladder (BCa) cancer are currently lacking. This study aimed to establish current practice patterns of PLND evaluation among pathologists. METHODS AND RESULTS: A web-based survey was circulated to all members of the European Network of Uropathology (ENUP), comprising 29 questions focusing on the macroscopic handling, LN enumeration and reporting of PLND in PCa and BCa. Two hundred and eighty responses were received from pathologists throughout 23 countries. Only LNs palpable at grossing were submitted by 58%, while 39% routinely embedded the entire specimen. Average LN yield from PLND was ≥10 LNs in 56% and <10 LNs in 44%. Serial section(s) and immunohistochemistry were routinely performed on LN blocks by 42% and <1% of respondents, respectively. To designate a LN microscopically, 91% required a capsule/subcapsular sinus. In pN+ cases, 72% reported the size of the largest metastatic deposit and 94% reported extranodal extension. Isolated tumour cells were interpreted as pN1 by 77%. Deposits identified in fat without associated lymphoid tissue were reported as tumour deposits (pN0) by 36% and replaced LNs (pN+) by 27%. LNs identified in periprostatic fat were included in the PLND LN count by 69%. CONCLUSION: This study highlights variations in practice with respect to the gross sampling and microscopic evaluation of PLND in urological malignancies. A consensus protocol may provide a framework for more consistent and standardised reporting of PLND specimens.

• Klíčová slova
• bladder cancer, lymph node, lymphadenectomy, prostate cancer,
• MeSH
• chirurgická patologie metody   normy   MeSH
• internet MeSH
• lidé MeSH
• lymfadenektomie * MeSH
• lymfatické metastázy * diagnóza   patologie   MeSH
• nádory močového měchýře * patologie   MeSH
• nádory prostaty * patologie   MeSH
• odběr biologického vzorku metody   normy   MeSH
• průzkumy a dotazníky MeSH
• staging nádorů metody   normy   MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Evropa MeSH

The polymicrobial biofilm (PMBF) is formed when microbes from multiple species co-aggregate into an envelope made of extra polymeric substances (EPS) that keep the microbes safe from external stresses. The formation of PMBF has been linked to a variety of human infections, including cystic fibrosis, dental caries, urinary tract infections, etc. Multiple microbial species co-aggregation during an infection results in a recalcitrant biofilm formation, which is a seriously threatening phenomenon. It is challenging to treat polymicrobial biofilms since they contain multiple microbes which show drug resistance to various antibiotics/antifungals. The present study discusses various approaches by which an antibiofilm compound works. Depending on their mode of action, antibiofilm compounds can block the adhesion of cells to one another, modify membranes/walls, or disrupt quorum-sensing systems.

• Klíčová slova
• Antibiofilm compounds, Antimicrobial resistance, Cell adhesion, Polymicrobial biofilm,
• MeSH
• antibakteriální látky farmakologie   MeSH
• antifungální látky MeSH
• biofilmy MeSH
• lidé MeSH
• quorum sensing MeSH
• zubní kaz * MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH
• Názvy látek
• antibakteriální látky MeSH
• antifungální látky MeSH