TRAP1 controls cell cycle G2-M transition through the regulation of CDK1 and MAD2 expression/ubiquitination.

Regulation of tumour cell proliferation by molecular chaperones is still a complex issue. Here, the role of the HSP90 molecular chaperone TRAP1 in cell cycle regulation was investigated in a wide range of human breast, colorectal, and lung carcinoma cell lines, and tumour specimens. TRAP1 modulates the expression and/or the ubiquitination of key cell cycle regulators through a dual mechanism: (i) transcriptional regulation of CDK1, CYCLIN B1, and MAD2, as suggested by gene expression profiling of TRAP1-silenced breast carcinoma cells; and (ii) post-transcriptional quality control of CDK1 and MAD2, being the ubiquitination of these two proteins enhanced upon TRAP1 down-regulation. Mechanistically, TRAP1 quality control on CDK1 is crucial for its regulation of mitotic entry, since TRAP1 interacts with CDK1 and prevents CDK1 ubiquitination in cooperation with the proteasome regulatory particle TBP7, this representing the limiting factor in TRAP1 regulation of the G2-M transition. Indeed, TRAP1 silencing results in enhanced CDK1 ubiquitination, lack of nuclear translocation of CDK1/cyclin B1 complex, and increased MAD2 degradation, whereas CDK1 forced up-regulation partially rescues low cyclin B1 and MAD2 levels and G2-M transit in a TRAP1-poor background. Consistently, the CDK1 inhibitor RO-3306 is less active in a TRAP1-high background. Finally, a significant correlation was observed between TRAP1 and Ki67, CDK1 and/or MAD2 expression in breast, colorectal, and lung human tumour specimens. This study represents the first evidence that TRAP1 is relevant in the control of the complex machinery that governs cell cycle progression and mitotic entry and provides a strong rationale to regard TRAP1 as a biomarker to select tumours with deregulated cell cycle progression and thus likely poorly responsive to novel cell cycle inhibitors. Copyright © 2017 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.

Bladder Metastases from Breast Cancer: Managing the Unexpected. A Systematic Review.

Breast cancer (BrC) has the highest incidence among females world over and it is one of the most common causes of death from cancer overall. Its high mortality is mostly due to its propensity to rapidly spread to other organs through lymphatic and blood vessels in spite of proper treatment. Bladder metastases from BrC are rare, with 50 cases having been reported in the last 60 years. This review aims to discuss some critical points regarding this uncommon condition. First, we performed a systematic review of the literature in order to draw a clinical and pathological profile of this entity. On this basis, its features in terms of diagnostic issues, imaging techniques, and survival are critically examined. Most bladder metastases from BrC are secondary lobular carcinoma, which mimic very closely the rare variant of urothelial cancer with lobular carcinoma-like features (uniform cells with an uncohesive single-cell, diffusely invasive growth pattern); thus, immunohistochemistry is mandatory to arrive at a correct diagnosis. This article summarizes the current knowledge regarding the incidence, clinical presentation, diagnosis, prognosis, and treatment of bladder metastases in patients with BrC.

Typing the atypical: Diagnostic issues and predictive markers in suspicious prostate lesions.

As much as 5% of prostate biopsies yield findings equivocal for malignancy even for skilled uropathologist; such "grey zone" lesions have been addressed in many ways, although the acronym ASAP (atypical small acinar proliferation) is the most widely used when referring to an atypical focus suspicious, but not diagnostic, for malignancy. Since the introduction of this diagnostic category more than 20 years ago, debate has ensued over its histological characterization and clinical significance. Pathology reporting of ASAP, commonly based on strict morphological criteria and traditional immunohistochemical markers such as basal cell antibodies, has been improved by recent availability of novel immunohistochemical markers such as AMACR and ERG. Further pathological issues, such as the role of pre-analytical variables, number of tissue levels, interobserver variability, and association with prostatic intraepithelial neoplasia also play a role in the optimal assessment of ASAP. Apart from diagnostic issues, a major issue is ASAP predictive value for prostate cancer on repeat biopsy. Therefore, attempts have been made to identify clinical and biological parameters that could predict subsequent diagnosis of malignancy as well as define time and modality of repeat biopsy. Finally, pathological features of cancers detected after a previous ASAP diagnosis are compared with those diagnosed at first prostate biopsy.

Bladder Metastases from Lung Cancer: Clinical and Pathological Implications: A Systematic Review.

Lung cancer is the tumor with the highest incidence in males worldwide and the most common cause of death from cancer overall; its high mortality is mostly due to its propensity to spread to other organs through lymphatic and blood vessels in spite of proper treatment. Bladder metastases from lung cancer are rare, with only 11 cases having been reported, all in recent years. This review aims to discuss some critical points regarding this uncommon condition, namely: (a) lung and bladder tumors share similar etiologic features; (b) almost all bladder metastases from lung cancer arise from lung adenocarcinomas; (c) cytology and superficial bladder biopsy may be falsely negative, since the neoplastic cells coming through the hematogenous route are typically located in the lamina propria and/or muscularis propria of the bladder wall; and (d) the differential diagnosis with primary bladder adenocarcinoma as well as primary and secondary small-cell carcinomas may be challenging. Though no definite conclusions can be drawn regarding treatment, we herein propose a practical algorithm to manage such patients based on available data.

Classification of stillbirths is an ongoing dilemma.

AIM: To compare different classification systems in a cohort of stillbirths undergoing a comprehensive workup; to establish whether a particular classification system is most suitable and useful in determining cause of death, purporting the lowest percentage of unexplained death. METHODS: Cases of stillbirth at gestational age 22-41 weeks occurring at the Department of Gynecology and Obstetrics of Foggia University during a 4 year period were collected. The World Health Organization (WHO) diagnosis of stillbirth was used. All the data collection was based on the recommendations of an Italian diagnostic workup for stillbirth. Two expert obstetricians reviewed all cases and classified causes according to five classification systems. RESULTS: Relevant Condition at Death (ReCoDe) and Causes Of Death and Associated Conditions (CODAC) classification systems performed best in retaining information. The ReCoDe system provided the lowest rate of unexplained stillbirth (14%) compared to de Galan-Roosen (16%), CODAC (16%), Tulip (18%), Wigglesworth (62%). CONCLUSION: Classification of stillbirth is influenced by the multiplicity of possible causes and factors related to fetal death. Fetal autopsy, placental histology and cytogenetic analysis are strongly recommended to have a complete diagnostic evaluation. Commonly employed classification systems performed differently in our experience, the most satisfactory being the ReCoDe. Given the rate of "unexplained" cases, none can be considered optimal and further efforts are necessary to work out a clinically useful system.

Molecular markers in bladder cancer: Novel research frontiers.

Bladder cancer (BC) is a heterogeneous disease encompassing distinct biologic features that lead to extremely different clinical behaviors. In the last 20 years, great efforts have been made to predict disease outcome and response to treatment by developing risk assessment calculators based on multiple standard clinical-pathological factors, as well as by testing several molecular markers. Unfortunately, risk assessment calculators alone fail to accurately assess a single patient's prognosis and response to different treatment options. Several molecular markers easily assessable by routine immunohistochemical techniques hold promise for becoming widely available and cost-effective tools for a more reliable risk assessment, but none have yet entered routine clinical practice. Current research is therefore moving towards (i) identifying novel molecular markers; (ii) testing old and new markers in homogeneous patients' populations receiving homogeneous treatments; (iii) generating a multimarker panel that could be easily, and thus routinely, used in clinical practice; (iv) developing novel risk assessment tools, possibly combining standard clinical-pathological factors with molecular markers. This review analyses the emerging body of literature concerning novel biomarkers, ranging from genetic changes to altered expression of a huge variety of molecules, potentially involved in BC outcome and response to treatment. Findings suggest that some of these indicators, such as serum circulating tumor cells and tissue mitochondrial DNA, seem to be easily assessable and provide reliable information. Other markers, such as the phosphoinositide-3-kinase (PI3K)/AKT (serine-threonine kinase)/mTOR (mammalian target of rapamycin) pathway and epigenetic changes in DNA methylation seem to not only have prognostic/predictive value but also, most importantly, represent valuable therapeutic targets. Finally, there is increasing evidence that the development of novel risk assessment tools combining standard clinical-pathological factors with molecular markers represents a major quest in managing this poorly predictable disease.

Predictive markers in bladder cancer: do we have molecular markers ready for clinical use?

Bladder cancer (BC) is a heterogeneous disease. Approximately 75% of patients present with non-muscle-invasive BC (NMIBC), which has a high recurrence rate and a low but unpredictable progression rate. Conversely, patients with muscle-invasive BC (MIBC) are at high risk for progression and cancer-specific mortality, but, again, disease behavior is unpredictable. To date, risk assessment for tumor recurrence and progression is based on clinico-pathological factors only. A risk assessment calculator that is based on several such parameters is available for NMIBC, but it has been reported to have potential flaws. In the last two decades, great effort has been made to evaluate the prognostic and predictive role of several molecular markers in MIBC and, even more so, in NMIBC, where the need for more precise risk stratification is urgently needed. This review addresses current evidence for the role of several molecular markers easily assessable by immunohistochemical techniques in prognosticating/predicting the outcome of NMIBC and MIBC. To date, because of divergent results among the many studies, no molecular marker has yet entered routine clinical practice; however, some of them (e.g., p53, pRb, p21, and survivin) have proved their predictive value in studies that included a homogeneous patient population on standardized treatment, and, therefore, are probably ready for clinical validation on a larger scale. Even more interesting is the possibility of constructing multimarker panels that could be used in routine clinical practice, as all these markers can easily be evaluated by immunohistochemistry on routine surgical pathology specimens. The molecular markers described herein hold promise for becoming widely available and cost-effective tools for reliable risk assessment, which would represent a great advancement in counseling patients, in selecting them for neoadjuvant and adjuvant treatments, and in determining their eligibility for clinical trials.

Prostate cancer detection rates in different biopsy schemes. Which cores for which patients?

PURPOSE: To determine whether the addition of four paramedian peripheral and four lateral peripheral cores improves the cancer detection rate (CDR) of the extended 10-core biopsy scheme and which patients benefit most from such additional samples. METHODS: One thousand and ninety-one consecutive patients scheduled for first ultrasound-guided transrectal prostate biopsy prospectively underwent a 18-core biopsy scheme, including the traditional sextant (6-core), 4 lateral peripheral (10-core), 4 paramedian peripheral (14-core) and additional 4 lateral peripheral cores (18-core). RESULTS: The CDR of the 6-, 10-, 14- and 18-core schemes was 33.1, 39.2, 41.6 and 41.8 %, respectively; the difference between the 10- and 6-core scheme reached significance (p < 0.005), whereas that between the 18- or 14- and the 10-core scheme did not. The percentage of tumors diagnosed on the sole basis of the 14-core scheme was significantly greater in patients with low PSA (≤ 7.2 vs. >7.2 ng/ml: 12.1 vs. 1.8 %; p < 0.0001), large prostate volume (≥ 50 vs. <50 cc: 3.4 vs. 9.1 %; p = 0.011) and particularly low PSA density (<0.15 vs. ≥ 0.15: 15.9 vs. 1 %; p < 0.0001). The 18-core scheme did not provide diagnostic advantages in any patients' population. CONCLUSIONS: The addition of 4 lateral peripheral samples did not increase the CDR of the 10-core biopsy scheme. The addition of four paramedian peripheral samples was beneficial only in patients with PSA density <0.15, in whom the 10-core scheme would have miss almost 16 % of tumors. Since more than half of our patients had low (<0.15) PSA density, these findings seem to be of great clinical relevance.

Bladder metastasis from lung adenocarcinoma: a difficult differential diagnosis with primary bladder adenocarcinoma.

BACKGROUND: Bladder metastases from lung adenocarcinoma are extremely rare; in the seven previously reported cases, the finding of an intact epithelium overlying the bladder tumour was considered suggestive of a secondary lesion. We describe the first case of bladder metastasis from lung adenocarcinoma whereby endoscopic appearance was strongly consistent with primary bladder cancer, thus complicating the differential diagnosis with primary bladder adenocarcinoma. CASE REPORT: A 65-year-old woman with a 13-year history of clean intermittent catheterization was diagnosed with a right lung adenocarcinoma metastatic to mediastinal and right supraclavicular nodes, as well as to the left lung, and treated with six cycles of cisplatin/pemetrexed, followed by six cycles of pemetrexed only. The 18-month follow-up computed tomography revealed several solid lesions of the bladder wall and she was scheduled for transurethral resection of bladder tumours. Endoscopic appearance was strongly consistent with primary bladder cancer but a thorough pathologic evaluation allowed the diagnosis of bladder metastasis from lung adenocarcinoma. CONCLUSIONS: Differentiating primary bladder adenocarcinoma from metastatic adenocarcinoma lesions can be difficult. An endoscopic appearance consistent with primary bladder cancer further complicates the differential diagnosis, which heavily relies on pathologic evaluation and specific immunohistochemical staining.

BRAF mutation and RASSF1A expression in thyroid carcinoma of southern Italy.

Aim of this work is to provide a detailed comparison of clinical-pathologic features between well-differentiated and poorly differentiated tumors according to their BRAF and RASSF1A status. We analyzed RASSF1A methylation by MSP and BRAF mutation by LCRT-PCR with LightMix® kit BRAF V600E in neoplastic thyroid tissues. Immunohistochemical evaluation of RASSF1A expression was also performed by standard automated LSAB-HRP technique. An overall higher degree of RASSF1A over-expression than normal thyroid parenchyma surrounding tumors (P < 0.05) has been found in all malignant well-differentiated lesions. Moreover, statistically significant higher levels of RASSF1A expression were observed in differentiated cancers associated to an inflammatory autoimmune background (P = 0.01). Amplifiable DNA for LC PCR with LightMix® kit BRAF V600E was obtained in nine PTCs, four FVPTCs, five ATCs, and one control. The V600E mutation was found in 13 of 18 (72%) tumors. BRAF was mutated in 6 of 9 (66%) classical PTC, in 2 of 4 (50%) follicular variant PTC and in all ACs (100%). The overall frequency of RASSF1A promoter methylation observed was 20.5% (9 cases out 44). Hypermethylation of RASSF1A in primary tumors was variable according to histotypes ranging from100% (5/5) in ACs to only 12.5% (4/32) in PTCs. We show a correlation between RASSF1A methylation status and RASSF1A protein expression. Finally, we conclude that BRAF V600E mutation and RASSF1A methylation were pathogenetic event restricted to a subgroup of PTC/FVPTCs in early stage and to clinically aggressive ATCs.

Combined perianal-intrarectal (PI) lidocaine-prilocaine (LP) cream and lidocaine-ketorolac gel provide better pain relief than combined PI LP cream and periprostatic nerve block during transrectal prostate biopsy.

UNLABELLED: What's known on the subject? and What does the study add? Study Type - Harm Reduction RCT Level of Evidence 1b The combination of perianal-intrarectal lidocaine-prilocaine cream and periprostatic nerve block effectively counteracts probe and sampling related pain during transrectal prostate biopsy, but not pain due to periprostatic infiltration. The novel combination of lidocaine-prilocaine cream and lidocaine-ketorolac gel, both administered perianal-intrarectally, provides the same probe and sampling-related pain relief than combined perianal-intrarectal lidocaine-prilocaine cream and periprostatic nerve block and prevents the non-negligible pain due to periprostatic infiltration, thus leading to better overall patients' compliance to the procedure. OBJECTIVE: • To compare the efficacy and safety of combined perianal-intrarectal (PI) lidocaine-prilocaine (LP) cream and lidocaine-ketorolac (LK) gel with combined PI LP cream and periprostatic nerve block (PPNB) in relieving pain during transrectal ultrasonography guided prostate biopsy (TPB). PATIENTS AND METHODS: • In all, 200 patients were randomized to receive combined PI LP cream and LK gel (group 1) or combined PI LP cream and PPNB (group 2) before TPB. • The 0-10-point visual analogue scale (VAS) was used for assessing pain at probe insertion and movements (VAS-1), periprostatic infiltration (VAS-2) when applied, and prostate sampling (VAS-3), as well as maximal procedural pain (MPP). • Complications occurring ≤ 20 days after the TPB were recorded. RESULTS: • The groups were comparable for patients' age, serum PSA level, prostate volume, and cancer detection rate. • All patients tolerated the procedure well. The two anaesthetic regimens provided almost equal mean VAS-1 (0.33 vs 0.37; P= 0.701) and VAS-3 (0.52 vs 0.51; P= 0.954) scores, but patients in group 2 reported significantly greater MPP scores (0.68 vs 1.53; P < 0.001) as periprostatic infiltration was the most painful part of the procedure (mean VAS-2: 1.33). • Complications rate was similar in the two groups (1% vs 2%; P= 0.38). CONCLUSIONS: • The novel combination of PI LP cream and LK gel provided the same probe- and sampling- related pain relief as combined PI LP and PPNB; moreover, by preventing the non-negligible periprostatic infiltration pain, it provided significantly better overall patients' compliance to the procedure. • Being safe and easy to administer, this novel non-infiltrative regimen has the potential to replace infiltrative anaesthesia in relieving pain during TPB.

Pelvic mass-like florid cystic endosalpingiosis of the uterus: a case report and a review of literature.

INTRODUCTION: Endosalpingiosis is a disorder of Mullerian system characterized by benign glands lined by tubal type epithelium and involves the peritoneum, subperitoneal tissues, and retroperitoneal lymph nodes. Endosalpingiosis is almost an incidental finding on microscopic examination. Seldom it appears as a cyst and it can be confused clinically for an ovarian tumor. DESIGN: A case report and a systematic review about pelvic mass-like florid endosalpingiosis of the uterus from PUB MED database were performed. PATIENT(S): We describe a case report of a 50-year-old woman with a pedunculated uterine neoformation, for which the preoperative exams could be compatible with an adnexal mass. Twelve patients with similar clinical history were discussed in the review. TREATMENT: Laparoscopy with radical exeresis was performed. RESULTS: Microscopic exam revealed florid cystic endosalpingiosis of the uterus. CONCLUSIONS: Endosalpingiosis is a rare mullerian disorder and the main problem is that the symptomatology is not specific and it may be initially misinterpreted. In relation to the papillary aspect of the lesion and focal calcifications, the histological differential diagnosis could include serous adenocarcinoma, but the lack of cellular stratification in absence of mitotic activity, and the presence of slight nuclear atypia contradict the diagnosis of carcinoma. The differential diagnosis for endosalpingiosis also includes multiple peritoneal inclusion cysts (benign cystic mesothelioma). The aim of this case report has not been only to describe the rarity of this pathology, but it contributes to consider endosalpingiosis as a possible diagnostic hypothesis for which may be indicated a conservative surgical treatment.

Expression and clinical implication of cyclooxygenase-2 and E-cadherin in oral squamous cell carcinomas.

Epithelial-Mesenchymal Transition (EMT) and angiogenesis are crucial events for development of aggressive and often fatal Oral Squamous Cell Carcinomas (OSCCs). Both promote cancer progression and metastasis development, but while the former induces the loss of E-cadherin expression and, hence cadherin switching; the latter produces hematic blood vessel neo-formation and contribute to OSCC cell growth, tumor mass development, and dissemination. Cyclooxygenase-2 (COX-2) has an important role, not only in angiogenic mechanisms, but also in favoring cancer invasion. Indeed it decreases the expression of E-cadherin and leads to phenotypic changes in epithelial cells (EMT) enhancing their carcinogenic potential. Our aim is to evaluate the interplay between E-cadherin cytoplasmic delocalization, COX-2 up-regulation and COX-2 induced neo-angiogenesis in 120 cases of OSCC. We have analyzed the distribution and the number of neo-formed endothelial buds surrounding infiltrating cells that express COX-2, as well as the neo-formed vessels in chronic inflammatory infiltrate, which surround the tumor. A double immunostaining method was employed in order to verify co-localization of endothelial cell marker (CD34) and COX-2. IHC has also been used to assess E-cadherin expression. Our data demonstrate that the OSCC cells, which lose membranous E-cadherin staining, acquiring a cytoplasmic delocalization, overexpress COX-2. Moreover, we find a new CD34+ vessel formation (sprouting angiogenesis). Only basaloid type of OSCC showes low level of COX-2 expression together with very low level of neo-angiogenesis and consequent tumor necrosis. The well-known anti-metastatic effect of certain COX-2 inhibitors suggests that these molecules might have clinical utility in the management of advanced cancers.

Survivin gene-expression and splicing isoforms in oral squamous cell carcinoma.

PURPOSE: Survivin, an inhibitor of apoptosis protein and a cell cycle regulator, has been detected in the majority of human cancers. Five splice variants (survivin, survivin-2alpha, survivin-2B, survivin-3B, and survivin-DeltaEx3) have been identified; their expressions have been investigated here. METHODS: By means of RT real-time PCR and immunohistochemistry, we have evaluated survivin isoform expressions at both mRNA and protein levels in human normal oral tissue, precancerous lesions, and oral squamous cell carcinoma (OSCC). Their correlations with the pathological findings have also been analyzed. RESULTS: Expression levels of all survivin transcript variants were markedly elevated in OSCC when compared to normal tissues. One-way analysis of variance (ANOVA) revealed highly significant up-regulation of survivin (P = 0.001), survivin-DeltaEx3 (P = 0.001) and survivin-2B (P = 0.004), whereas survivin-3B showed a minor increase in OSCC compared to normal mucosa. CONCLUSIONS: Our findings suggest that survivin isoforms deregulation may have significant implications in tumor aggressiveness and prognosis.

Urethral corpus spongiosum amyloidosis presenting with urethrorrhagia during erection.

INTRODUCTION: Urethral amyloidosis is a rare, probably inflammatory condition usually presenting with hematuria and obstructive urinary symptoms, thus mimicking urethral malignancy. After histological confirmation of the diagnosis, treatment can be expectant or symptomatic. AIM. To report an unusual cause of urethrorrhagia occurring only during erection in an otherwise healthy man. METHODS. A 30-year-old man presented with a 5-month history of urethrorrhagia occurring only during erection, and with a painless palpable nodule in his penile urethra clearly visible on urethral US and magnetic resonance imaging, but not on urethroscopy. RESULTS. The patient underwent wide surgical excision of the urethral nodule and grafting of the urethral defect with a pedicled preputial flap. Histological examination revealed isolated amyloid of urethral corpus spongiosum. CONCLUSIONS. Isolated urethrorrhagia during erection and without urinary symptoms can be the presenting sign of urethral amyloidosis involving corpus spongiosum rather than the urethral lumen; in such cases, surgical exploration, wide urethral excision and grafting are mandatory.

A case of ovarian psammocarcinoma associated with endometrioid cysts: a morphological and immunohistochemical study.

BACKGROUND: Psammocarcinoma of the ovary is an uncommon neoplasm, currently classified among the low-grade epithelial serous tumors. Little is known about its pathogenesis and biological behavior. CASE: A 35-year-old woman underwent laparotomic myomectomy and surgical removal of the right gonad after the incidental discovery of an adnexal mass. The pathological findings were consistent with serous psammocarcinoma and multiple endometrioid cysts of the ovary. Tissue specimens from the neoplasm were tested by immunohistochemistry for the p53, HER-2/neu and bcl-2 cancer-related proteins; diffuse overexpression for bcl-2 was detected, while tumor cells were negative for p53 and HER-2/neu. CONCLUSION: Although the pathogenesis and clinical course of ovarian psammocarcinoma are still to be determined, the molecular profile of this case highlights the similarities with ovarian tumors of the serous borderline group.

Survivin expression in renal cell carcinoma.

Deregulated expression of inhibitors of apoptosis may contribute to cancer by aberrantly extending cell viability and facilitating the insurgence of resistance to chemo- and radiotherapy. In this study, we have investigated, by immunohistochemical technique, the expression and potential prognostic significance of survivin in a series of 49 clear cell type renal cell carcinoma (ccRCC). Survivin expression was significantly associated with poorly differentiated, advanced stages and more aggressive ccRCCs (p < 0.05). Patients with low survivin expression had statistically significant better survival rates than patients with high survivin expression (p < 0.05). This may be relevant for follow-up protocols design and/or alternative therapeutic approaches.

Altered expression of HER-2 and the mismatch repair genes MLH1 and MSH2 predicts the outcome of T1 high-grade bladder cancer.

PURPOSE: The identification of factors predicting the outcome of stage T1 high-grade bladder cancer (BC) is a major clinical issue. METHODS: We performed immunohistochemistry to assess the role of human epidermal growth factor receptor-2 (HER-2) and microsatellite instability (MSI) factors MutL homologue 1 (MLH1) and MutS homologue 2 (MSH2) in predicting recurrence and progression of T1 high-grade BCs having undergone transurethral resection of bladder tumor (TURBT) alone or TURBT + intravesical instillations of bacillus Calmette-Guerin (BCG). RESULTS: HER-2 overexpression was a significant predictor of disease-free survival (DFS) in the overall as well as in the two patients' population; as for progression-free survival (PFS), it was significant in the overall but not in the two patients' population. MLH1 was an independent predictor of PFS only in patients treated with BCG and MSH2 failed to predict DFS and PFS in all populations. Most importantly, the higher the number of altered markers the lowers the DFS and PFS. In multivariate Cox proportional-hazards regression analysis, the number of altered molecular markers and BCG treatment were significant predictors (p = 0.0004 and 0.0283, respectively) of DFS, whereas the number of altered molecular markers was the only significant predictor (p = 0.0054) of PFS. CONCLUSIONS: Altered expression of the proto-oncogene HER-2 and the two molecular markers of genetic instability MLH1 and MSH2 predicted T1 high-grade BC outcome with the higher the number of altered markers the lower the DFS and PFS. These findings provide grounds for further testing them in predicting the outcome of this challenging disease.

Urine profiling by SELDI-TOF/MS: monitoring of the critical steps in sample collection, handling and analysis.

The topic of this study is the impact of several pre-analytical and analytical variables on proteomic profiling of human urine by surface enhanced laser desorption/ionization time of flight-mass spectrometry (SELDI-TOF-MS) in healthy subjects. Urine storage at room temperature caused a progressive degradation of proteins, which was prevented by the addition of protease inhibitors only up to 2 h from the collection. The timing of collection over the day had only a minor impact on protein profile, although influencing the intensity of peaks. Repeated freeze/thaw cycles (up to five) did not affect either the number or the intensity of the peaks. A comparison of the protein profile from eight different healthy individuals showed fairly consistent inter-subject similarities, along with between-subject differences, which were markedly dependent on the sex and the type of ProteinChip array used. The addition of a variety of denaturing agents improved the quality of the spectra with all the chips tested (CM10, Q10 and H50), but not with the copper-coated IMAC-30 chip. Finally, SPA matrix allowed to achieve a better performance of SELDI-TOF/MS spectrum, as compared with CHCA, regardless of the ProteinChip array used and even in the low m/z range (2500-10,000). In conclusion, we suggest that a careful choice of a number of pre-analytical and analytical conditions is required to accomplish and define a unifying protocol for the analysis of human urine by SELDI-TOF/MS, in physiological and in pathological states.

Mitochondrial dysfunctions in bladder cancer: Exploring their role as disease markers and potential therapeutic targets.

Bladder cancer (BC) is a major cause of mortality worldwide as it currently lacks fully reliable markers of disease outcome and effective molecular targets for therapy. Mitochondria play a key role in cell metabolism but the role of mitochondrial dysfunctions in BC has been scarcely investigated. In this review, we explored current evidence for the potential role of mitochondrial DNA (mtDNA) alterations (point mutations and copy number) as disease markers in BC. Some germline mtDNA mutations detectable in blood could represent a non-invasive tool to predict the risk of developing BC. MtDNA copy number and tumor specific mtDNA mutations and RNAs showed encouraging results as novel molecular markers for early detection of BC in body fluids. Moreover, mitochondrial proteins Lon protease, Mitofusin-2, and TFAM may have prognostic/predictive value and may represent potential therapeutic targets. A deeper understanding of mitochondrial dysfunctions in BC could therefore provide novel opportunities for targeted therapeutic strategies.