Silver-stained Nucleolar Organizer Regions Research Articles

A Cytotaxonomic Survey of the Genus Melanophryniscus Gallardo, 1961 (Anura: Bufonidae)

Abstract We present a cytogenetic survey of the basal bufonid genus Melanophryniscus that covered 14 of the 25 species currently recognized, representing the three phenetic species groups: M. moreirae, M. stelzneri, and M. tumifrons. All species presented a diploid chromosome complement constituted by 11 bi-armed chromosome pairs (2n = 2x = 22; FN = 44). Some remarkable differences were observed between species groups: chromosome pair 4 was metacentric in species of the M. tumifrons group (also with a distinctive C-positive block) but submetacentric in the M. stelzneri group and M. sanmartini (M. moreirae group); pair 5 was submetacentric in M. sanmartini and metacentric in the rest. Chromosome secondary constrictions and silver-stained nucleolar organizer regions were located either in pair 5, 7, or 8 in the M. tumifrons group, M. sanmartini and M. krauczuki (M. stelzneri group), and M. stelzneri group, respectively; and pair 7 was relatively larger in M. sanmartini and M. krauczuki. Studied cytoge...

Parathyroid carcinoma presenting as normocalcemic hyperparathyroidism

Parathyroid carcinoma (PC) is a rare malignancy, with an indolent but progressive course. Long-term survival is largely dependent on the extent of the primary surgical resection. Hence, pre- or intraoperative suspicion of malignancy is of great importance. We describe the case of a 62-year-old woman with a 2-year history of asthenia and mental depression. Her past medical history was significant for osteoporosis. A diagnosis of primary normocalcemic hyperparathyroidism was established and the patient underwent surgery. PC was suspected intraoperatively because of the size and appearance of the parathyroid mass (a grayish, lobulated 3.5 cm mass). Thus, aggressive surgery (en bloc resection) was performed, along with bilateral neck exploration. Pathological examination of the specimens confirmed the suspicion of PC, demonstrating vascular invasion and extracapsular infiltration into adjacent soft tissue. Immunohistochemical staining revealed an elevated Ki-67 score (8.43%; cut-off value 5%). The mean area of silver-stained nucleolar organizer regions (AgNOR) was high (4.972 μm(2)), indicating an elevated proliferation rate. Serum calcium and parathyroid hormone levels normalized postoperatively, and the patient's 5-year outcome was good. The present case provides evidence that parathyroid malignancy cannot be excluded a priori based on normocalcemic hyperparathyroidism, emphasizing the variability in clinical presentation. Moreover, Ki-67 expression and AgNOR analysis confirmed their additional value in complementing the histological evaluation of a parathyroid malignant mass.

Quantification of AgNOR expression in exfoliated oral mucosal cells of tobacco chewers with and without lesion

Nucleolar organizer regions (NOR) are associated with proliferative activity and represent a diagnostic and prognostic marker. Smears were taken from smokers, tobacco chewers, oral squamous cell carcinoma patients, and normal subjects and evaluated by 2 silver-staining nucleolar organizer region (AgNOR) counting methods: (1) mean number of AgNORs per nucleus (mAgNOR); and (2) percentage of nuclei with >3 and >5 AgNORs (pAgNOR). A statistically significant difference was observed between normal subjects, smokers, tobacco chewers, and oral cancer patients and between tobacco chewers with and without lesion. No significant difference was observed between tobacco chewers and smokers except in the percentage of >5 criteria. AgNOR enumeration using noninvasive methods, such as the cytobrush appears to be useful technique in distinguishing between normal mucosa, mucosa with and without lesions exposed to carcinogens, such as tobacco and frank oral carcinoma.

Role of proliferative markers in breast lesions

Carcinoma of breast is a leading cause of death in women throughout the world. Difficulty in diagnosis occurs in routine Hematoxylin and Eosin (H&E) stained sections to differentiate premalignant lesions like epitheliosis and malignant lesions like carcinoma in situ which may affect prognosis. Our study was done to assess the role of proliferative markers in this field. In this study, total 66 cases of different breast lesions were studied by H&E stained sections as well as proliferative markers like silver staining nucleolar organizer region (AgNOR) and proliferating cell nuclear antigen (PCNA). Statistically significant differences found in expression of proliferative markers between most of them. Study of proliferative markers help in the proper diagnosis of different breast lesions which lies in the grey zone on routine histopathology.

Occurrence of natural triploidy in Rhamdia quelen (Siluriformes, Heptapteridae)

Five specimens of Rhamdia quelen collected from the Lindóia Stream, PR, Brazil, were cytogenetically analyzed. The diploid chromosome number found was 58, including 30 metacentric, 16 submetacentric, 10 subtelocentric, and 2 acrocentric chromosomes. Supernumerary or B chromosomes, frequently observed in this fish group, were not detected. One of the individuals was triploid, with 3n = 87. A silver-stained nucleolar organizer region was found on a pair of submetacentric chromosomes of the diploid specimens, and on three chromosomes of the triploid individual, confirming triploidy. Treatment with fluorochrome chromomycin A(3) revealed fluorescent bands coincident with those of the silver-stained nucleolar organizer region, in both diploid and triploid individuals, showing that this is a GC-rich region. Heterochromatin distribution was visualized by the C-banding technique, mainly in the terminal chromosome regions of the individuals and was also observed in the pericentromeric regions of some chromosomes and at both telomeres.

Nucleolin on the Cell Surface as a New Molecular Target for Gastric Cancer Treatment

Nucleolin is an abundant non-ribosomal protein found in nucleolus and a major component of silver-stained nucleolar organizer region (AgNOR), a histopathological marker of cancer which is highly elevated in cancer cells. We recently reported that nucleolin on the cell surface of mouse gastric cancer cells acts as a receptor for tumor necrosis factor-alpha-inducing protein (Tipalpha), a new carcinogenic factor of Helicobacter pylori. In this study, we first examined the localization of nucleolin on cell surface of five gastric cancer cell lines by cell fractionation and flow cytometry: We found that large amounts of nucleolin were present on surface of MKN-45, KATOIII, MKN-74, and AGS cells, with smaller amounts on surface of MKN-1 cells. The membrane fraction of normal epithelial cells of mouse glandular stomach did not contain much nucleolin, suggesting that translocation of nucleolin to the cell surface occurs during carcinogenesis, making for easier binding with Tipalpha. AS1411, a nucleolin targeted DNA aptamer, inhibited growth of gastric cancer cell lines in this order of potency: MKN-45>KATOIII>AGS>MKN-74=MKN-1, associated with induction of S-phase cell cycle arrest. Fluorescein isothiocyanate (FITC)-AS1411 was more rapidly incorporated into MKN-45 and AGS than into MKN-1 cells, based on varying amounts of cell surface nucleolin. We think that AS1411 first binds to nucleolin on the cell surface and that the binding complex is then incorporated into the cells. All results indicate that nucleolin on the cell surface is a new and promising therapeutic target for treatment of gastric cancer.

Comparative chromosome mapping of repetitive sequences. Implications for genomic evolution in the fish, Hoplias malabaricus

BackgroundSeven karyomorphs of the fish, Hoplias malabaricus (A-G) were previously included in two major groups, Group I (A, B, C, D) and Group II (E, F, G), based on their similar karyotype structure. In this paper, karyomorphs from Group I were analyzed by means of distinct chromosomal markers, including silver-stained nucleolar organizer regions (Ag-NORs) and chromosomal location of repetitive sequences (18S and 5S rDNA, and satellite 5SHindIII-DNA), through fluorescence in situ hybridization (FISH), in order to evaluate the evolutionary relationships among them.ResultsThe results showed that several chromosomal markers had conserved location in the four karyomorphs. In addition, some other markers were only conserved in corresponding chromosomes of karyomorphs A-B and C-D. These data therefore reinforced and confirmed the proposed grouping of karyomorphs A-D in Group I and highlight a closer relationship between karyomorphs A-B and C-D. Moreover, the mapping pattern of some markers on some autosomes and on the chromosomes of the XY and X1X2Y systems provided new evidence concerning the possible origin of the sex chromosomes.ConclusionThe in situ investigation of repetitive DNA sequences adds new informative characters useful in comparative genomics at chromosomal level and provides insights into the evolutionary relationships among Hoplias malabaricus karyomorphs.

The impact of sodium chloride on root growth, cell division, and interphase silver-stained nucleolar organizer regions (AgNORs) in root tip cells of Allium cepa L.

To evaluate genetic damage, parameters including root growth, chromosomal aberrations, and interphase silver-stained nucleolar organizer regions (AgNORs) in root tip cells of Allium cepa L. were investigated after treatment with NaCl (40–160 mM). The results showed that NaCl caused a decrease in root growth. All concentrations of NaCl showed an inhibitory effect on dividing cells in root tips of A. cepa L. and caused a reduction in mitotic index values. Upon exposure to NaCl, roots exhibited various mitotic abnormalities, including c-mitosis, anaphase bridge, and chromosome stickiness. In addition, interphase cells with micronuclei, budding nuclei, and unequal-sized nuclei were observed. Moreover, total cell aberration increased with increasing NaCl concentration. For AgNOR parameters, the average number of AgNORs per nucleus decreased in roots treated at all NaCl concentrations. The singular AgNOR area and whole AgNOR area in the nucleus containing 1–3 AgNORs were inversely proportional to NaCl concentrations.

Digital Image Analysis of Mean AgNOR Area Objectively Discriminates between Reactive Plasmacytosis and Multiple Myeloma in Cytologic Smears from Bone Marrow Aspirates

Objective: To differentiate between reactive plasmacytosis (RP) and multiple myeloma (MM) in bone marrow aspirates by assessment of silver-stainable nucleolar organizer regions (AgNORs). Methods: Archival cytologic smears (n = 69) from bone marrow aspirates were retrospectively investigated. Twenty cases had RP and 49 cases MM (24 low-grade, 22 intermediate-grade and 3 high-grade MM). Diagnoses were confirmed by evaluation of the corresponding bone marrow trephine biopsies. Cytologic smears were subjected to AgNOR staining using a standardized silver-staining protocol and assessed by two independent clinical histopathologists using digital image analysis. AgNOR count and area were analyzed in 100 plasma cell nuclei per sample. Results: The mean AgNOR area was found to be significantly (p < 0.001) higher in low-grade MM (mean: 23.4 µm²; range: 19.2–26.7 µm²) than in RP cases (mean: 6.2 µm²; range: 3.8–8.4 µm²). In contrast, mean AgNOR count failed to disclose significant differences between low-grade MM (mean: 5.36; range: 1–7) and RP (mean: 3.68; range: 1–6). Conclusion: Digital image analysis of the mean AgNOR area might be used as a diagnostically useful adjunct to aid objectively in the challenging differentiation between RP and MM in bone marrow aspirates.

Sequential demonstration of nucleolar organizer regions and Ki67 immunolabelling in non-Hodgkin's lymphomas

In order to investigate the relationship between the numbers of interphase silver-stained nucleolar organizer regions (AgNORs) and cell proliferation, we have studied prospectively a small series of non-Hodgkin's lymphomas (NHL). We applied the recently described double staining technique for sequential immunostaining and the AgNOR reaction to the same sections of tumour from each case. The immunostaining was performed with the antibody Ki67, known to demonstrate the nuclei of proliferating cells. A significant difference was found between the AgNOR scores in Ki67+ and Ki67- cells, the counts being much higher in the former than the latter. This was the case for both high- and low-grade NHL and is further evidence that interphase AgNOR counts reflect cell proliferation level, at least in lymphomas.

Cytogenetic analysis of the Asian Plethodontid salamander, Karsenia koreana: Evidence for karyotypic conservation, chromosome repatterning, and genome size evolution

A cytogenetic analysis, including the karyotype, C-bands, silver-stained nucleolus organizer regions and genome size, was performed on the recently discovered species, Karsenia koreana, the first plethodontid salamander from Asia. The karyotype consists of 14 pairs of bi-armed chromosomes, with no evidence of heteromorphic sex chromosomes. C-banding reveals a concentration of heterochromatin at the centromeres as well as at interstitial locations. The smallest chromosome (pair number 14) has symmetrical interstitial C-bands in each arm, resembling chromosome no. 14 of North American species of its sister group taxon, supergenus Hydromantes. Acomparative analysis of C-band heterochromatin and silver-stained nucleolus organizer regions of Karsenia and other plethodontid genera reveals that chromosomal evolution may have featured chromosome 'repatterning' within the context of conserved chromosome number and shape in this clade. Genome size is correlated with geographic distribution in plethodontids and appears to have important phenotypic correlates as well. The genome size of Karsenia is relatively large, and resembles that of the geographically closest plethodontids from western North America, especially species of the genus Hydromantes. The biological significance of these cytogenetic characteristics of plethodontid salamanders is discussed within an evolutionary context.

Evaluation of AgNOR scores in aspiration cytology smears of breast tumors

<b>Aim: </b> The aim of this study is to assess the difference in the silver-stained nucleolar organizer regions (AgNOR) scores on aspiration smears from benign and malignant breast tumors, and to determine the feasibility of AgNOR staining as part of the cytological investigation of breast tumors. <br><b> Settings: </b> AgNOR scores were evaluated on aspiration smears and tissue sections from 42 breast tumor cases.<br><b> Materials and Methods:</b> Smears from 27 breast tumor cases and tissue sections from 15 surgically resected specimens were examined. Counting was performed on a small number of tumor cells (50 cells). Both the mean (mAgNOR) score and the AgNOR proliferative index (pAgNOR index) were assessed for each specimen.<br><b> Statistical Analysis:</b> Statistical evaluation was carried out using the Students' 't'-test. The result was considered significant when <i>P</i> < 0.05. <br><b> Results and Conclusions:</b> The mAgNOR score and pAgNOR index were significantly higher in smears from malignant tumors than in those from benign tumors (<i>P</i> = 0.01). These results were similar to those obtained for tissue sections and were comparable to established data. The determination of AgNOR scores on cell smears in a limited cell population has practical advantages.

The diagnostic and prognostic implications of silver-binding nucleolar organizer regions in periodontal lesions

The periodontal lesions with cellular proliferation can be assessed by various methods. One of the most recent methods to determine the proliferative activity is silver-staining nucleolar organizer region (AgNOR) staining. The purpose of the present study was to evaluate, if AgNOR count can act as a proliferative marker and can aid in the diagnosis and prognosis of periodontal lesions. For this study, subjects with healthy gingival status, non-neoplastic lesions, neoplastic lesions, and plaque-induced gingivitis were included. Following the provisional diagnosis of clinical entity, biopsies were taken from the respective selected sites for histopathological diagnosis. In plaque-induced gingivitis cases, a second biopsy was taken from the selected sites 3 weeks following scaling. After histological confirmation, one more section was prepared, which was subjected to AgNOR staining, and AgNOR numbers were counted by individual and cluster counts and statistically analyzed. Results showed the highest AgNOR count in neoplastic lesions. Non-neoplastic lesions showed a higher AgNOR count as compared to clinically healthy gingiva. Plaque-induced gingivitis showed a considerable reduction in AgNOR count after treatment. Results of this study confirmed that AgNOR count reflects the cellular proliferation and has a limited diagnostic value. However, the prognostic value of AgNOR for periodontal lesions is dependable.

Silver-stained nucleolar organizer regions and immunoglobulins in cutaneous squamocellular tumors

This study was conducted to elucidate the biological activity of epidermal cells in cutaneous squamocellular tumors by counting the number of silver-stained nucleolar organizer regions (AgNORs), to estimate the quantity of Ig-producing cells and the inflammatory cellular infiltrate (ICI), and to achieve a comparative evaluation. Twenty cases of actinic keratosis (AK), 20 in situ squamous cell carcinomas (ISC), and 20 invasive squamous cell carcinomas (SCCs) were assessed using a silver colloid technique. Ig-producing or binding cells and ICI were also investigated immunohistochemically. In all samples, AgNORs, Ig-producing cells, and ICI increased in proportion to the degree of malignancy. With regard to AgNORs values, a statistically significant difference was confirmed between AK and ISC ( p<0.01), AK and SCC ( p<0.001), and ISC and SCC ( p<0.05). IgG-producing cells predominated in each case. Furthermore, a linear correlation was detected between ICI and AgNORs in AK and ISC. The significant difference in the number of AgNORs among the 3 stages of involution of SCC reinforces the value of AgNORs as a marker for malignant potential. Despite the absence of a correlation between AgNORs and the proportion of Ig-producing cells, the association between ICI and AgNORs in AK and ISC was obvious.

Silver-Stained Nucleolar Organizer Regions in Normal and Dysplastic Cervical Lesions: Correlation with DNA Ploidy and S-Phase Fraction by Flow Cytometry

Objectives: To investigate the prognostic significance of DNA ploidy and silver-stained nucleolar organizer regions (AgNOR), their relation to the cytological grades of cervical dysplasia and correlation of cells in the S-phase of the cell cycle and DNA ploidy with AgNOR count. Methods: Multiparametric DNA flow-cytometric analysis and AgNOR count were performed on cervical smear samples, cytologically diagnosed as normal (n = 196), atypical squamous cells of unknown significance (ASCUS, n = 98) and various grades of cervical dysplasia (n = 127). Results: Among the cytological grade, aneuploid cases from low-grade squamous intraepithelial lesion (mild) and high-grade squamous intraepithelial lesion (moderate and severe) groups were showing high AgNOR counts (ranges 4.33–5.86 and 5.91–7.42, respectively). Aneuploid cases from the ASCUS group also presented high AgNOR (range 3.72–4.93). The high percentage of cells in the S-phase (>12%) correlated with these 2 parameters in the ASCUS and dysplasia groups. After grouping the cases into those with high (>4.0) and low (<4.0) AgNOR counts, significant correlations in the group with high AgNOR count with DNA ploidy, S-phase and cytological grade were recorded (p < 0.001). Results obtained from follow-up study revealed that cases with aneuploidy, high AgNOR count and S-phase fraction presented with a progressive trend of disease. Conclusions: Thus, the study shows that high AgNOR count, S-phase fraction and aneuploid DNA have prognostic significance in the early detection of cervical cancer.

Computer-assisted analysis of cell proliferation markers in oral lesions

Abnormalities in any component of the cell cycle regulatory machine may result in oral cancer, and markers of cell proliferation have been used to determine the prognosis of tumor progression. The aim of this study was to determine whether silver-stained nucleolar organizer region (AgNOR) and Ki-67 measurements could improve the assessment of growth rates in oral lesions. Eighty-three oral biopsies were studied, 20 of which were classified as fibrous inflammatory hyperplasia (FIH), 40 as leukoplakia (LKP) and 23 as oral squamous cell carcinoma (OSCC). Within the LKP group, 22 out of 29 biopsies were diagnosed as non-dysplastic leukoplakia (LK) and 18 as dysplastic leukoplakia (DLK), presenting discrete, moderate and severe dysplasia. Ki-67 immunolabeling of the lesions increased steadily in the following order: FIH, DLK, LK and OSCC, indicating that Ki-67 is a good marker for predicting the proliferative fraction among benign, premalignant and malignant oral lesions. The median values of AgNOR parameters indicate that the morphometric index gives better results regarding the proliferative rate than the numerical one. A series of linear regressions between AgNOR parameters and Ki-67 showed positive associations. We conclude that a combination of Ki-67 and morphometric AgNOR analyses could be used as an aid in the determination of the proliferative status of oral epithelial cells in oral cancer.

Silver-stained organizer regions and immunoglobulins in cutaneous keratoacanthomas and squamous cell carcinomas

The aim of this study was to investigate the biologic activity of epidermal cells in keratoacanthomas (KAs) and squamous cell carcinomas (SCCs) by counting the number of silver-stained nucleolar organizer regions (AgNORs), to estimate the quantity of Ig-producing cells and the inflammatory cellular infiltrate (ICI), and to make a comparative evaluation. Thirty KAs (10 at growth stage, 10 at mature stage, and 10 at involution stage) and 28 SCCs (nine well differentiated-Grade 1 (G1), seven moderately differentiated-Grade 2 (G2), five poorly differentiated-Grade 3 (G3), and seven pseudoadenoid) were investigated. The KAs examined had a mean number of 1.727 AgNORs (S.D. 0.232), and IgG predominated in most cases. IgG and IgE increase at the involution, IgA remains at almost the same level, and IgM decreases during the maturity stage. The SCCs examined had a mean number of 2.105 AgNORs (S.D. 0.446). IgG predominated and gradually increased in proportion to the degree of malignancy. There is a significant difference in the number of AgNORs and the proportion of Ig subclasses in contrast to the cellular infiltrate among the three stages of KA. In SCCs, the number of AgNORs and the percentage of Igs and ICI increased gradually in proportion to the degree of malignancy.

Aurora-B Regulates RNA Methyltransferase NSUN2

Disassembly of the nucleolus during mitosis is driven by phosphorylation of nucleolar proteins. RNA processing stops until completion of nucleolar reformation in G(1) phase. Here, we describe the RNA methyltransferase NSUN2, a novel substrate of Aurora-B that contains an NOL1/NOP2/sun domain. NSUN2 was concentrated in the nucleolus during interphase and was distributed in the perichromosome and cytoplasm during mitosis. Aurora-B phosphorylated NSUN2 at Ser139. Nucleolar proteins NPM1/nucleophosmin/B23 and nucleolin/C23 were associated with NSUN2 during interphase. In mitotic cells, association between NPM1 and NSUN2 was inhibited, but NSUN2-S139A was constitutively associated with NPM1. The Aurora inhibitor Hesperadin induced association of NSUN2 with NPM1 even in mitosis, despite the silver staining nucleolar organizer region disassembly. In vitro methylation experiments revealed that the Aurora-B-phosphorylation and the phosphorylation-mimic mutation (S139E) suppressed methyltransferase activities of NSUN2. These results indicate that Aurora-B participates to regulate the assembly of nucleolar RNA-processing machinery and the RNA methyltransferase activity of NSUN2 via phosphorylation at Ser139 during mitosis.

Divergent location of ribosomal genes in chromosomes of fish thorny-headed worms, Pomphorhynchus laevis and Pomphorhynchus tereticollis (Acanthocephala)

We studied distribution of ribosomal DNA (rDNA) sequences along with chromosomal location of the nucleolar organizer regions (NORs) in males of two fish parasites, Pomphorhynchus laevis and Pomphorhynchus tereticollis (Acanthocephala). Fluorescence in situ hybridization with 18S rDNA probe identified two clusters of rDNA in each species, but revealed a remarkable difference in their location on chromosomes. In P. laevis, the rDNA-FISH signals were found in long arms of the first chromosome pair and in short arms of the second pair. Whereas in P. tereticollis, rDNA clusters were located in long arms of both the first and second chromosome pairs. The divergent location of rDNA clusters in the chromosome No. 2 supports current classification of P. tereticollis, previously considered a synonym of P. laevis, as a separate species. A possible scenario of the second chromosome rearrangement during karyotype evolution of the two species involves two successive pericentric inversions. In both species, one or two prominent nucleoli were apparent within interphase nuclei stained with either silver nitrate or a fluorescent dye YOYO-1. However, a single large nucleolus was observed in early stages of mitosis and meiosis I regardless the number of rDNA clusters. Nevertheless, two bivalents with silver-stained NORs in diakinesis and two silver-stained sites in early prophase II nuclei indicated that all NORs are active. This means that each Pomphorhynchus NOR generates a nucleolus, but the resulting nucleoli have a strong tendency to associate in a large body.

Chemoprevention of mouse urinary bladder carcinogenesis by fermented brown rice and rice bran

Fermented brown rice by Aspergillus oryzae (FBRA) has been shown to be a potent anti-carcinogenic compound. Here, we investigated the modifying effects of dietary feeding with a naturally occurring anti-oxidant FBRA on N-butyl-N-(4-hydroxybutyl)-nitrosamine (OH-BBN)-induced urinary bladder carcinogenesis in male ICR mice. Five-week-old male ICR mice were divided into 7 groups, and groups 1-5 were given OH-BBN (500 ppm) in drinking water for 6 weeks starting at 7 weeks of age. Groups 2 and 3 were fed the diet containing 5% and 10% FBRA during the initiation phase, respectively, whereas groups 4 and 5 were fed these diets during the post-initiation phase. Group 6 was given the diet containing 10% FBRA throughout the experiment, and group 7 was kept on the basal diet alone and served as an untreated control. At the end of the study (week 32), the incidences of simple hyperplasia, dysplasia and carcinoma in the bladders of group 1 (OH-BBN alone) were 92%, 49% and 38%, respectively. Those of group 5 (64%, 23% and 10%) and the incidence of carcinoma of group 4 (17%) was significantly less than that of group 1. Furthermore, the multiplicity of simple hyperplasia and carcinoma of group 5 was significantly less than that of group 1. Post-initiation exposure of 10% FBRA significantly decreased the number/nucleus of silver-stained nucleolar organizer region proteins (AgNORs), an index of cell proliferation, in the non-lesional transitional epithelium when compared to that of the control. Our results indicate that FBRA exerts chemopreventive effects against chemically induced urinary bladder carcinogenesis through anti-proliferative mechanisms. FBRA could be a promising chemopreventive agent for human urinary bladder cancer.