Urinary Bladder Specimens Research Articles

Malignant lymphoma of the lower urinary tract: A single institutional experience

Lymphoma of the urinary tract is relatively rare and comprises of < 5% of all primary extra nodal lymphoma. Diagnoses of these lesions at anearly stage is important as they can disseminate or transform into high grade lesion if there is a delay in the diagnoses. There are only few case series and case reports on the malignant lymphoma of the urinary tract. The aim of this study was to characterize lymphoma involving the urinary bladder and prostate. We retrospectively reviewed the clinical data and histologic findings of the malignant lymphoma involving urinary bladder and prostate at our institution. Lymphoma involving the lower urinary tract clinically presented with lower urinary tract symptoms and usually with concurrent associated urinary bladder cancer or prostatic cancer in our series. Lymphoma should be included in the differential diagnoses especially in patients with prior history of lymphoid disorders. There should be a high index of suspicion when there is any atypical lymphoid infiltrate in routine urinary bladder and prostate surgical specimens.

Cytoprotective Antioxidant, Anti-Inflammatory, and Antifibrotic Impact of Celery Seed Oil and Manuka Honey Against Cyclophosphamide-Induced Cystitis in Rabbits.

Patients treated with cyclophosphamide (CP) usually suffer from severe hemorrhagic cystitis (HC). Our previous study exhibited that mesna + celery cotherapy partially ameliorated HC. Therefore, there is a substantial need to seek alternative regimens to get complete protection against CP-induced HC. The current study investigated the effects of mesna + celery seed oil (MCSO) or mesna + manuka honey (MMH) cotherapy against CP-induced HC in adult male rabbits. The forty rabbits were divided into four equal groups and treated for three weeks. The control group (G1) received distilled water and the second group (G2) received CP (50 mg/kg/week). The third group (G3) received CP + MCSO (CPMCSO regimen), and the fourth group (G4) received CP + MMH (CPMMH regimen). The urinary bladder (UB) specimens were processed to evaluate UB changes through histopathological, immunohistochemical, ultrastructural, and biochemical investigations. In G2, CP provoked HC features (urothelial necrosis, ulceration, and sloughing), UB fibrosis, and TNF-α immunoexpression. Besides, CP reduced the activity of antioxidant enzymes (GPx1, SOD3, and CAT) and elevated the serum levels of NF-κB, TNF-α, IL-1B, and IL-6 cytokines in G2 rabbits. In contrast, the CPMMH regimen caused significant increments of UB protection against HC in G4 rabbits compared to the partial protection by the CPMCSO regimen in G3. Therefore, our study indicated for the first time that the novel CPMMH regimen resulted in complete UB protection against CP-induced HC via combined antioxidant, anti-inflammatory, and antifibrotic properties.

Effect of Tadalafil on Apoptosis and Proliferation in Urinary Bladder Mucosa after Chemically Induced Haemorrhagic Cystitis; A Histological and Immunohistochemical Study

Introduction: Haemorrhagic cystitis (HC) can be chemically induced by cyclophosphamide (CYP) injection; it is mainly used as an immunosuppressive and antineoplastic drug. Tadalafil (TAD) a phosphodiesterase type-5 inhibitor has vasodilator, anti-inflammatory and anti-apoptotic effects. It is mainly used for the treatment of erectile dysfunction and pulmonary arterial hypertension.Aim of the work: This work aimed to verify the effect of tadalafil on apoptosis and proliferation on the injured bladder mucosa after chemically induced HC. Material and Methods: Thirty-six adult male albino rats were randomly divided into four groups; group I was the control group, group II was TAD-treated group, group III was CYP-treated group and group IV was TAD-CYP treated group. Groups III and IV were subdivided into two subgroups (a and b), subgroups IIIa and IVa were euthanized after one day while subgroups IIIb and IVb were euthanized after 10 days. Urinary bladder specimens were processed for light and electron microscopic examination and immunohistochemical analysis of caspase-3 and Ki-67. Results: Subgroup IIIa revealed haemorrhage and decrease of urothelium thickness with presence of denuded areas. Subgroup IIIb showed restoration of urothelium with focal denuded areas. Both subgroups showed a significant increase in apoptotic index while subgroup IIIb showed a significant increase in proliferative index. Subgroup IVa revealed a significant decrease in apoptosis associated with a significant increase in proliferation, thereby limiting cellular loss. Subgroup IVb; showed a significant decrease in both apoptosis and proliferation indicated prevention of the developed hyperplasia of the urothelium. By transmission electron microscopy, the terminal differentiation of superficial cells of group IV was expressed by presence of an angular contour.Conclusion: TAD-CYP co-treatment provides a positive impact on the apoptosis, proliferation, and the histological changes of HC on urinary bladder mucosa. Therefore, further studies are required to elucidate the protective effect of TAD on CYP-induced HC.

Identification of six novel alternative transcripts of the human kallikrein-related peptidase 15 (KLK15), using 3'RACE and high-throughput sequencing.

The kallikrein-related peptidase 15 (KLK15) gene is a member of the largest cluster of serine proteases in the human genome. Exhibiting trypsin-like activity, KLK15 is most likely involved in the activation of prostate-specific antigen (PSA; also known as KLK3), an established biomarker for the diagnosis and screening of prostate cancer. High mRNA expression levels of KLK15 have already been reported in ovarian and prostate cancer, in contrast with breast cancer, where KLK15 has been proposed as a biomarker of favorable prognosis. In this study, we exploited the next-generation sequencing (NGS) technology along with 3' rapid amplification of cDNA ends (3' RACE) to discover alternative KLK15 splice variants. Extensive computational analysis of the obtained NGS data revealed the existence of novel splice junctions, thus supporting the existence of novel KLK15 transcripts. Six novel KLK15 splice variants were identified and verified by Sanger sequencing. Two of them (KLK15 v.11 and v.12) contain an open reading frame and are hence predicted to encode two novel KLK15 protein isoforms. Expression analysis of each KLK15 splice variant in sixteen cDNA pools from malignant cell lines and in normal cell lines (HEK293, HaCaT, and BJ cells) revealed very different expression profiles of particular KLK15 transcripts. Moreover, the new KLK15 splice variants were shown to be expressed in breast, ovarian, prostate, urinary bladder, colon, and renal tissue specimens. Due to the prominent clinical value of KLK15 mRNA expression, the novel KLK15 transcripts appear as candidate cancer biomarkers for diagnostic and/or prognostic purposes and, therefore, merit further investigation.

Intimate Attachment of Escherichia coli O157:H7 to Urinary Bladder Epithelium in the Gnotobiotic Piglet Model.

Enterohemorrhagic Escherichia coli (EHEC), a pathogenic subset of Shiga toxin-producing E. coli (STEC), is an important cause of hemorrhagic colitis and hemolytic–uremic syndrome (HUS), and a rare cause of urinary tract infections (UTIs) with associated HUS. EHEC strains attach intimately to intestinal epithelium with formation of actin pedestals (attaching-effacing (A/E) lesions); however, the mechanism of EHEC attachment to the uroepithelium is unknown. We conducted a retrospective study on archived urinary bladder specimens from gnotobiotic piglets that naturally developed cystitis associated with EHEC O157:H7 infection following oral inoculation and fecal shedding. Paraffin-embedded bladder tissues from three piglets with cystitis and immunohistochemical evidence of EHEC O157:H7 adherence to the uroepithelium were processed for and examined by transmission electron microscopy. EHEC O157:H7 bacteria were found in one of three piglets, intimately attached to pedestals on the apical surfaces of the superficial urothelium (umbrella cells). Cystitis was significantly associated with the length of survival of the piglets post-inoculation (p = 0.0339; estimated odds ratio = 2.6652). This is the first report of E. coli causing A/E-like lesions in the uroepithelium, and also evidence of the utility of the gnotobiotic piglet as a model for studies of the pathogenesis of EHEC UTIs.

The applicability and utility of immunohistochemical biomarkers in bladder pathology

Urinary bladder specimens are frequently encountered in the daily practice of surgical pathologists. The spectrum of pathologic entities encountered in bladder specimens is extraordinarily broad, and in some instances, immunohistochemical stains are used to help characterize challenging bladder lesions. Cost-effective biomarker selection tailored to the differential diagnosis facilitates an accurate diagnosis. This comprehensive review is prepared as a reference guide for the use of immunohistochemistry to categorize primary and secondary bladder neoplasms and to evaluate metastatic cancers for possible bladder origin.

Preparation and Utilization of Freshly Isolated Human Detrusor Smooth Muscle Cells for Characterization of 9-Phenanthrol-Sensitive Cation Currents.

Detrusor smooth muscle (DSM) cells present within the urinary bladder wall ultimately facilitate urine storage and voiding. Preparation of the viable, fresh, and isolated DSM cells presents an important technical challenge whose achievement provides optimal cells for subsequent functional and molecular studies. The method developed and elaborated herein, successfully used by our group for over a decade, describes dissection of human urinary bladder specimens obtained from open bladder surgeries followed by an enzymatic two-step treatment of DSM pieces and mechanical trituration to obtain freshly isolated DSM cells. The initial step involves dissection to separate the DSM layer (also known as muscularis propria) from mucosa (urothelium, lamina propria, and muscularis mucosa) and the adjacent connective, vascular, and adipose tissues present. The DSM is then cut into pieces (2–3 mm x 4–6 mm) in nominal Ca2+-containing dissection/digestion solution (DS). DSM pieces are next transferred to and sequentially treated separately with DS containing papain and collagenase at ~37 °C for 30–45 min per step. Following washes with DS containing enzyme-free bovine serum and trituration with a fire-polished pipette, the pieces release single DSM cells. Freshly isolated DSM cells are ideally suited for patch-clamp electrophysiological and pharmacological characterizations of ion channels. Specifically, we show that the TRPM4 channel blocker 9-phenanthrol reduces voltage-step evoked cation currents recorded with the amphotericin-B perforated patch-clamp approach. DSM cells can also be studied by other techniques such as single cell RT-PCR, microarray analysis, immunocytochemistry, in situ proximity ligation assay, and Ca2+ imaging. The main advantage of utilizing single DSM cells is that the observations made relate directly to single cell characteristics revealed. Studies of freshly isolated human DSM cells have provided important insights characterizing the properties of various ion channels including cation-permeable in the urinary bladder and will continue as a gold standard in elucidating DSM cellular properties and regulatory mechanisms.

Analysis of Intestinal Metaplasia Without Dysplasia in the Urinary Bladder Reveal Only Rare Mutations Associated With Colorectal Adenocarcinoma.

Intestinal metaplasia (IM) is a rare finding in urinary bladder specimens. It is unclear whether IM without dysplasia is a precursor of malignancy in the urinary system. We retrospectively selected 9 cases of IM of bladder (1 case harboring high-grade dysplasia), and performed mutation analysis for genes frequently mutated in colon cancer including BRAF, APC, KRAS, MET, NRAS, PIK3CA, CTNNB1, FBXW7, and TP53 using validated clinical tests. Control groups included 7 colonic tubular adenomas, 10 high-grade papillary urothelial carcinomas. One IM case revealed an APC mutation and another showed an NRAS mutation. Among the tubular adenomas cases, 6 of 7 (85.7%) harbored KRAS mutations and 3 of 7 (42%) APC mutations. Among urothelial carcinomas cases, 1 revealed a KRAS mutation, 2 had PIK3CA mutations, and all cases were negative for APC mutations. Clinical follow-up for the IM patients was available with a median follow-up of 70 months. One patient-without any mutation in the genes investigated-developed invasive bladder adenocarcinoma with intestinal differentiation with metastasis to the liver and lung. Neither of the 2 patients harboring mutations developed any malignancy. In conclusion, a minority of cases with IM without dysplasia bear mutations in the genes commonly associated with colonic adenocarcinoma, suggesting a premalignant potential for such lesions possibly following the classic multistep chromosomal instability pathway of carcinogenesis. A larger cohort of patients with longer follow-up is needed to better establish whether close follow-up is warranted for mutation-harboring IM of the bladder.

A histological analysis of urinary bladder specimens with elaboration of various neoplastic lesions

A histological analysis of urinary bladder specimens with elaboration of various neoplastic lesions - IJPO- Print ISSN No: - 2394-6784 Online ISSN No:- 2394-6792 Article DOI No:- 10.18231/j.ijpo.2019.130, Indian Journal of Pathology and Oncology-Indian J Pathol Oncol

Plasmacytoid/diffuse urothelial carcinoma: a single-institution immunohistochemical and molecular study of 69 patients

Accurate diagnosis of plasmacytoid urothelial carcinoma (PUC) is important given its poor prognosis and frequent presentation at high stage. We aim to assess the clinicopathological features, molecular aberrations, and follow-up data in a series of PUC cases from a single tertiary cancer center. Seventy-two urinary bladder, ureteral, and renal pelvic specimens with urothelial carcinoma with plasmacytoid differentiation were identified. Immunohistochemical stains were performed on 48 cases. Among urinary bladder origin markers, GATA3 was most sensitive (96%). Breast carcinoma markers (estrogen receptor, mammaglobin) were usually negative, but progesterone receptor stained 1 case (4%). Neuroendocrine markers CD56 and TTF-1 were each positive in 1 case (4% and 4%, respectively). Gastrointestinal adenocarcinoma marker CDX2 was positive in 4 cases (15%), but nuclear β-catenin was negative in all cases. CD138 was positive in 83% and E-cadherin expression was lost in 57% of cases. Fluorescence in situ hybridization using the UroVysion Bladder Cancer Kit and FGFR3 mutational analysis using polymerase chain reaction were performed on 15 cases; deletion of chromosome 9p21 was common (60%), and FGFR3 mutations were detected in 60% of cases (5 cases had both deletion 9p21 and FGFR3 mutations). Cases were divided into 3 morphologic groups: classic (29%), desmoplastic (35%), and pleomorphic (36%). The 3 morphologic subtypes had distinct survival outcomes (P = .083), with median survival for all patients 18 being months versus 10 months for the desmoplastic group.

Utility of Additional Tissue Sections in Surgical Pathology.

To avoid diagnostic errors such as missed diagnosis and errors in staging tumors due to inadequate tissue sampling, pathologists submit additional sections (AS). This study assessed frequency, diagnostic yield, distribution, and cost of AS. Among 1542 AS cases, we calculated mean AS per case; fraction of AS that altered diagnosis or stage; AS variation by tissue, malignant versus benign lesions, presence or absence of neoadjuvant therapy, mass, margin, lymph nodes, or other source, resident versus pathologist assistant (PA) dissector; and AS cost per case. Overall 9.2 ± 8.8 AS were collected per case. In only 3.8% (58/1542) of cases AS altered diagnosis or stage. Urinary bladder cases provoked the most AS: 19.5 ± 15.1 per case. Significantly more AS came from malignant versus benign lesions (10.8 ± 9.7 vs 7.6 ± 7.5, P = <.0001) and from specimens treated with neoadjuvant therapy versus malignant lesions not so treated (12.3 ± 9.4 vs 10.3 ± 9.8, P = .02). Lymph nodes were sampled more heavily compared with mass, margin, and other sites combined (11.8 ± 11.4 vs 8.9 ± 8.4, P = .003), but in 78.4% (1209/1542) of cases, AS were from mass. Of diagnosis or stage altering AS cases, two thirds (38/58) were from masses, one fifth (11/58) from lymph nodes, a 10th (6/58) from margins, and a 20th (3/58) from other specimen sites. Resident versus pathologist assistant dissection caused no significant AS difference. AS contributed 40% cost per case. AS per case ranged widely; their diagnostic yield was low; they were highest in urinary bladder specimens, in malignant and particularly neoadjuvant-treated lesions. Although lymph nodes were most heavily sampled, most AS were from masses. Resident dissection did not increase AS and cost of AS was high.

CK20 and p53 Immunohistochemical Staining Patterns in Urinary Bladder Specimens With Equivocal Atypia.

- Urinary bladder flat carcinoma in situ (CIS) is a worrisome lesion, requiring aggressive surveillance and treatment. Cytokeratin 20 (CK20) and p53 are common immunohistochemical antibodies used to supplement CIS diagnosis in biopsy samples. However, existing data come primarily from unequivocally benign and malignant specimens. - To correlate these markers in specimens with borderline histology with outcomes. - CK20 and p53 immunohistochemistry was analyzed for staining pattern, classified as CIS pattern (both stains yielding strong labeling of the area of concern), discordant (only 1 stain yielding CIS pattern), indeterminate (1 or both stains yielding partial or equivocal labeling), or benign (both stains yielding a benign pattern). - Specimens with equivocal atypia (n = 69) from 65 patients were studied. There were 9 specimens (13%) that had a CIS staining pattern, 18 (26%) were discordant, 31 (45%) were indeterminate, and 11 (16%) were benign. Of the discordant specimens, 13 labeled for CK20 but not p53, whereas 5 showed the opposite. Most specimens (n = 47; 68%) were obtained from patients with a known history of bladder cancer, of which recurrence developed in 27, with an average interval of 37 months (range, 2-216 months). A subset (n = 22; 34%) had no prior history of bladder cancer, from which only 1 patient with CK20-positive/p53-equivocal staining later developed diagnostic carcinoma. - In our cohort of specimens with equivocal urothelial atypia, very few patients without a prior diagnosis of bladder cancer progressed to diagnostic cancer (1 of 22), suggesting that staining results should be interpreted with caution in de novo atypia. Patients with a known history of bladder cancer had a substantial rate of recurrence, independent of staining pattern.

Experimental model for acute kidney injury caused by uropathogenic Escherichia coli.

Acute kidney injury (AKI) is the rapid deterioration of renal function, diagnosed on the basis of an increase in serum creatinine and abnormal urinary parameters. AKI is associated with increased risk of mortality or chronic kidney disease (CKD). The aim of the study was to develop an experimental model for AKI resulting from Escherichia coli-induced pyelonephritis. E. coli was isolated from a patient with clinical symptoms of urinary tract infection (UTI). The study included three groups of female Wistar rats (groups 1, 2 and 3), in which pyelonephritis was induced by transurethral inoculation with highly virulent E. coli (105, 107 and 109 cfu/ml, respectively). Urine and blood samples for analysis were obtained prior to the inoculation (day 0), as well as 7, 14 and 21 days thereafter. Aside from a microbiological examination of urine samples, daily urine output, serum creatinine (CreaS), creatinine clearance (CrCl), interleukin 6 (IL-6), fractional excretion of sodium (FENa) and fractional excretion of urea (FEUrea) were determined. A histopathological examination of kidney and urinary bladder specimens was conducted as well. While UTI-related pyelonephritis developed irrespective of E. coli inoculum size, AKI was observed only following transurethral administration of E. coli at the intermediate and high dose, i.e. 107 and 109 cfu/ml, respectively (group 2 and 3). An increase in CreaS and abnormal diuresis were accompanied by changes in parameters specific for various forms of AKI, i.e. FENa and FEUrea. Based on these changes, administration of E. coli at 107 cfu/ml was demonstrated to induce renal AKI, whereas inoculation with 109 cfu/ml seemed to cause not only ascending pyelonephritis, but perhaps also bacteremia and urosepsis (prerenal component of AKI).

The diagnostic role of vimentin in differentiation between muscularis propria and muscularis mucosa in urinary bladder carcinoma

Objective The aim of this study was to evaluate the diagnostic value of vimentin expression for differentiation between muscularis mucosa (MM) and muscularis propria (MP) in urinary bladder carcinoma cases. Background The single most important prognostic factor in urothelial carcinoma is the pathological stage, which includes the anatomic depth of invasion on which major therapeutic decisions are made. Correct assessment of invasion necessitates discrimination between MM and MP. Vimentin is the most widely distributed intermediate filament and is expressed in virtually all mesenchymal tissues. Materials and methods This study was carried out on 70 urinary bladder specimens, including 59 cases of primary urinary bladder carcinoma and 11 cystitis specimens. The specimens were submitted to vimentin immunostaining under the streptavidin–biotin amplified system. Results MM was identified histologically in 40 cases, whereas MP was morphologically designated in all cases (70 cases). Vimentin was negative in MP and showed positive expression in 32/40 (80%) cases of MM with a statistically significant difference (P = 0.000), 80% of sensitivity, and 100% of specificity. Conclusion Negative vimentin expression in MP could be a useful tool for identification of MP with 80% of sensitivity and 100% of specificity in urinary bladder carcinoma cases.

Expression of 5-Hydroxytryptamine Receptors in Human Urinary Bladders with Benign Prostatic Hyperplasia

This study investigated the mRNA expression pattern and distribution of 5-hydroxytryptamine (5-HT) receptors 5-HT2A, 5-HT2B, 5-HT3A, 5-HT4, and 5-HT7 within the urothelium and detrusor of normal bladder tissue and in the urothelium of bladders from patients with benign prostatic hyperplasia (BPH). Normal urinary bladder specimens were obtained from 13 patients undergoing radical cystectomy due to bladder cancer (normal group) and BPH specimens were obtained from 27 benign prostatic obstruction patients receiving transurethral prostatectomy or retropubic prostatectomy. Receptor subtype mRNA expression was determined by real-time reverse transcription polymerase chain reaction on urothelium, detrusor, and whole mucosal preparations. Receptor distribution was determined by immunohistochemistry. In normal tissues, expressions of 5-HT2B and 5-HT7 receptor mRNAs in the urothelium, detrusor, and whole mucosa were greater than the average expression for all receptor subtype mRNAs. 5-HT2B receptor protein was distributed in the apical urothelium and among the detrusor smooth muscle layers. In contrast, the 5-HT7 receptors were within the urothelium middle cell layers and detrusor smooth muscle cells. The expression pattern of each 5-HT receptor subtype mRNA within the BPH urothelium was similar to that in the normal urothelium. The expression level of 5-HT2A receptor mRNA in the BPH group was significantly lower than the normal group; however, the expressions of both 5-HT3A and 5-HT7 mRNAs were significantly higher. The expressions of both 5-HT2B and 5-HT4 mRNAs were not significantly different between the normal and BPH groups. In normal urinary bladders, the expressions of both 5-HT2B and 5-HT7 mRNAs were higher compared to the 5-HT2A, 5-HT3A, and 5-HT4 mRNAs. The distributions of 5-HT2B and 5-HT7 receptors were different in the urothelium and detrusor layers. The 5-HT3A and 5-HT7 receptor mRNAs in the BPH group were significantly higher compared to the normal urothelium, while the 5-HT2A mRNA was significantly lower. Asahi Kasei Pharma Corporation.

Role of dexamethasone alone and in combination with quercetin in the urinary bladder after administration of ifosfamide

Background Ifosfamide (IFO) is a broad-spectrum antineoplastic agent. Dexamethasone inhibits the activity of a variety of cytokines. Quercetin is a flavonoid with an antioxidant effect. Aim This work aimed to study the possible protective role of dexamethasone alone and in combination with quercetin on the IFO-treated urinary bladder. Materials and methods Sixty adult albino rats were used in this study and were divided into six groups: group I was the control group; group II was the dexamethasone-treated group; group III was the dexamethasone and quercetin-treated group; group IV was the IFO-treated group in which each rat was injected with IFO; group V was the IFO and dexamethasone-treated group, in which each rat was injected with IFO and three doses of dexamethasone; and group VI was the IFO, dexamethasone and quercetin-treated group, in which each rat was injected with IFO and three doses of dexamethasone and quercetin. Urinary bladder specimens were processed for histological and immunohistochemical staining for inducible nitric oxide synthase and nuclear factor-κB (NF-κB), followed by different morphometric examinations. Results Group IV revealed areas of epithelial denudation and vacuolated cells. Group V revealed partially restored thickness of the urothelium with vacuolated cells. Group VI showed restored epithelial integrity with reduced cytoplasmic vacuoles. Electron microscopic examination of the urothelium revealed indented nuclei and dilated intercellular spaces in group IV. After administration of dexamethasone, indented nuclei and partially dilated intercellular space were seen. In group IV, the urothelium revealed apparently normal nuclei with almost intact intercellular junctions. Highly significant increase in mast cell count in group IV but nonsignificant increase after administration of both dexamethasone and quercetin were noticed. Optical density of inducible nitric oxide synthase and NF-κB immunoreaction was highly significantly increased in group IV. Significant increase was detected in group V for NF-κB but nonsignificant increase in group VI for both immunohistochemical stains was observed. Conclusion It could be concluded that combined dexamethasone and quercetin exerted better protective roles against the toxic effects induced by IFO rather than dexamethasone alone.

Bladder cancer: Analysis of the 2004 WHO classification in conjunction with pathological and geographic variables

Bladder cancer (BCA) is a worldwide disease and shows a wide range of geographical variation. The aim of this study is to analyze the prevalence of schistosomal and non-schistosomal associated BCA as well as compare our findings with the 2004 WHO consensus classification of urothelial neoplasms and with other publications. The archival materials of 180 urinary bladder specimens were collected from Department of Pathology, King Abdul-Aziz University Hospital (KAUH), Jeddah, Western region, Saudi Arabia. The regional prevalence of this cancer was identified and studied, and a comparison between schistosomal and non-schistosomal associated BCA was made. Additionally, the study revised and classified these neoplasms according to the most recent 2004 WHO classification of urothelial neoplasms. The type of mural invasion either muscularis mucosae (MM) or muscularis propria (MP), other associated lesions as carcinoma in situ (CIS) as well as, metaplasia and schistosomal infestation were assessed. Urothelial cell neoplasms (UCN) comprised 161 cases (89.4%), squamous cell carcinoma (SCC) represented 12 cases (6.7%), adenocarcinoma 5 cases (2.8%), and sarcomatoid carcinoma detected in 2 cases (1.1%). Among all these cases schistosomal associated BCA represented 13 cases (7.2%), while the remaining 167 cases (92.8%) were non-schistosomal associated BCA. In the former category, 11 cases (6.1%) were squamous cell carcinoma and 2 (1.1%) urothelial cell carcinoma (UCC) whereas, non-schistosomal associated cancer that included UCN, SCC, adenocarcinoma, and sarcomatoid carcinoma found in 159, one case, 5, and 2 cases. Invasion of muscularis propria was detected in 30 cases (16.7%) and muscularis mucosae invasion in 45 cases (25%). According to WHO classification of urothelial neoplasms accurate categorization of BCA is very important for both diagnostic and therapeutic values.

Expression of Oct4A splicing variant in human bladder premalignant lesions predicts its invasiveness

Stem cells are immature, unspecialized cells that are able to proliferate and population self-renewal. In tumor mass, the cells with properties similar to stem cells are present. These cells, so-called cancer stem cells (CSC), are small subpopulation of cells, which can initiate the tumor and cause the recurrence of the tumor. According to published data, CSC have different level of sensitivity to chemo- and radiotherapy, dependent, i. e. on its differentiation state. It seems that complete recovery of the patients is possible only when all CSC will be eliminated. There are many of stem cell markers. In our research we analyzed the expression of OCT4A splicing variant of OCT4 (POU5F1) gene in preinvasive and malignant lesions of the bladder. OCT4A is a marker of embrional stem cells and, as transcription factor, is responsible for sustaining the cells in undifferentiated state, characteristic of the stem cell and potential to self-renewal. Our previous study suggested that its expression is observed in premalignant and malignant lesions and are consistent with results of other authors. We examine archival, buffered formalin-fixed, paraffin-embedded tissue samples of urinary bladder specimens obtained from 113 patients. Expression of OCT4A was detected using immunohistochemistry. The intensity of OCT4A immunostaining was scored as 0 (negative), 1 (faint) or 2 (distinct) and OCT4A-positive cells were analyzed as the numbers of cells per 1000 tumor cells. Based on our results, we found the most intensive and the most extensive (measured by means of percentage of OCT4A-positive cells) expression of this marker in urothelial cells at in situ stage. In papillary tumors (pTa) and in situ cancers the percentage of OCT4A-positive cells was higher when compared to less advanced invasive cancers (pT1 and pT2), whereas the percentage of OCT4A-positive cells in pTa tumors was similar to that observed in invasive tumors at pT4 stage. The observed differences were well-defined and statistically significant. To summarize, the highest number and the most intensive staining of OCT4A-positive cells in non-invasive lesions pTis and pTa could suggest the important role of OCT4A in initiation and promotion of invasive process. It cannot be excluded that, analysis and evaluation of the expression of this transcription factor can have predictive value in the prediction of progression of pre-invasive tumor to invasive form.

Infiltration of Epstein‐Barr virus‐harboring lymphocytes occurs in a large subset of bladder cancers

Epstein-Barr virus (EBV) has been implicated in the genesis of a variety of human cancers. We aimed to confirm the presence and define the role of EBV in bladder cancer. A total of 39 bladder cancer specimens were analyzed. Ten urinary bladder tissues obtained at autopsy were used as a normal control. EBV-encoded RNA (EBER) was evaluated by in situ hybridization (ISH). Frozen material available from 18 EBER-positive cases was analyzed by using reverse transcription-polymerase chain reaction for BZLF1, an early lytic gene product. The expression of CD20, CD3, ZEBRA (BZLF1 product) and transforming growth factor beta1 (TGFbeta-1) was assessed using an immunohistochemical technique. Infiltration of EBER-expressing lymphocytes was detected in 26 of 39 bladder cancer cases (66.7%). A small fraction of the tumor cells as well as the infiltrating lymphocytes were positive in two cases. All normal urinary bladder specimens showed negative results. The incidence of EBV-positive lymphocyte infiltration was significantly higher for advanced stage cancers than those in earlier stages (Ta-152% vs T2-4 93%, P = 0.013). The presence of BZLF1 mRNA was demonstrated in seven out of the 18 EBER-positive cases. Infiltration of EBV-harboring lymphocytes occurs in a large subset of bladder cancers cases. It is more frequently associated with advanced stages. EBV infection in tumor cells is very limited. Our findings suggest that EBV-positive lymphocytes might play a role in bladder cancer progression.

Telomere Shortening and Chromosomal Abnormalities in Intestinal Metaplasia of the Urinary Bladder

Although intestinal metaplasia is often found in association with adenocarcinoma of the urinary bladder, it is unclear whether intestinal metaplasia of the bladder is a premalignant lesion. Telomere shortening has recently been implicated in epithelial carcinogenesis. We used quantitative fluorescent in situ hybridization (FISH) to measure telomere length and UroVysion FISH to detect cytogenetic abnormalities in urinary bladder specimens with intestinal metaplasia. Paraffin-embedded tissue blocks from 34 patients with intestinal metaplasia of the urinary bladder were evaluated. Twelve of the 34 patients had coexistent cystitis glandularis, and telomere length was measured in these lesions for comparison. Tissue sections were prepared and hybridized with a telomere-specific peptide nucleic acid probe. Quantitative FISH on interphase nuclei was used to assess telomere signal intensity. Additional sections were hybridized with centromeric probes for chromosomes 3, 7, and 17 and a locus-specific probe 9p21. Multicolor FISH was used to analyze for cytogenic abnormalities in the interphase nuclei of intestinal metaplasia. In all 34 cases, reduced average telomere signal intensity was observed in the nuclei of intestinal metaplasia cells compared with adjacent control nuclei to produce a mean relative intensity of 48.5% (P < 0.0001). When cystitis glandularis was present, significant differences in the telomere-specific signal intensity existed between cystitis glandularis and normal cells (P = 0.0005) and between cystitis glandularis and intestinal metaplasia cells (P = 0.0015). Three of the 34 cases showed chromosomal gains in the UroVysion FISH assay. Our findings indicate that intestinal metaplasia in the urinary bladder is associated with significant telomere shortening relative to telomere length in adjacent normal urothelial cells. These lesions also occasionally showed cytogenetic abnormalities associated with telomere shortening. Our findings support the hypothesis that intestinal metaplasia of the urinary bladder is a precursor lesion to and could be a marker in the development of adenocarcinoma of the urinary bladder.