Predictors and prognostic implications of clinical decisions in patients with primary high-risk non-muscle-invasive bladder cancer - results of a cross country retrospective study.

Adjuvant diagnostic and therapeutic procedures are available to reduce the risk of recurrence or progression in patients with high-risk non-muscle-invasive bladder cancer (NMIBC). However, their indications and efficacy remain a matter of debate. The aim of this study was to analyze therapeutic decisions in patients with primary high-risk NMIBC and to analyze the adherence to clinical guidelines in this field.545 consecutive patients, aged a median of 70.3 years, diagnosed with primary high-risk NMIBC in thirteen urological institutions, were enrolled into this retrospective study. Diagnostic and therapeutic decisions after transurethral resection (TUR) were recorded, and predictive factors were analyzed.Restaging TUR was offered to 260 patients (47.7%), up-front intravesical Bacillus Calmette-Guerin (BCG) therapy to 74 patients (13.6%), immediate radical cystectomy to 38 patients (7.0%), and intravesical chemotherapy with the maintenance therapy to 12 patients (2.2%). No additional procedure was performed in 161 patients (29.5%). The strongest predictive factor for restaging TUR was G3 or high-grade cancer (RR 1.68, p<0.01), for upfront BCG therapy it was carcinoma in situ (RR 3.20, p=0.01), for immediate cystectomy it was stage T1 tumor (RR 3.71, p<0.01), for no additional procedures it was G2 or low-grade cancer (RR 2.18, p<0.01).Clinical management of patients with high-risk NMIBC is suboptimal and not standardized. As this can directly influence patients' survival, urgent improvement of urological care in this field should be considered.

Stage of bladder cancer in Central Europe - Polish perspective.

Mortality rate from bladder cancer in Europe is the highest in its Central Region. This study is an attempt to find underlying factors by proper characterisation of large cohort of Polish patients with bladder cancer.This is a multicentre study enrolling 1360 consecutive patients diagnosed with primary urothelial carcinoma of the bladder in years 2012-2013 in Poland. All patients underwent transurethral resection of the bladder tumor. Data on staging and grading of all cancers were collected, as well as several demographic and clinical factors were tested for the association with muscle invasiveness of the cancer.Mean age of the cohort was 69.6 years, male to female ratio was 3:1. Bladder cancer stage Ta, T1 and muscle-invasive (MIBC) was diagnosed in 533 (39.2%), 516 (37.9%) and 296 (21.8%) patients, respectively. Patients with MIBC were older (73 vs. 68 years, p<0.05), had lower body mass index (25.4 vs. 26.5 kg/m2, p<0.05), lower haemoglobin concentration (12.2 vs. 13.4 mg/l, p<0.05), longer history of haematuria (86.2 vs. 74.4 days) and longer time interval from first symptom to diagnosis (118.0 vs. 88.2 days), compared to patients with Ta and T1 tumors.High mortality rate from bladder cancer in Central Europe can result from very high incidence of high-risk T1 tumors and high prevalence of prognostic factors of poor survival.

The ClpX heat-shock protein of Escherichia coli, the ATP-dependent substrate specificity component of the ClpP-ClpX protease, is a novel molecular chaperone.

All major classes of protein chaperones, including DnaK (the Hsp70 eukaryotic equivalent) and GroEL (the Hsp60 eukaryotic equivalent) have been found in Escherichia coli. Molecular chaperones enhance the yields of correctly folded polypeptides by preventing aggregation and even by disaggregating certain protein aggregates. Previously, we identified the ClpX heat-shock protein of E. coli because it enables the ClpP catalytic protease to degrade the bacteriophage lambda O replication protein. Here we report that ClpX alone possesses all the properties expected of a molecular chaperone protein. Specifically, it can protect the lambda O protein from heat-induced aggregation, disaggregate preformed lambda O aggregates, and even promote efficient binding of lambda O to its DNA recognition sequence. A lambda O-ClpX specific protein-protein interaction can be detected either by a modified ELISA assay or through the stimulation of ClpX's weak ATPase activity by lambda O. Unlike the behaviour of the major DnaK and GroEL chaperones, ClpX requires the presence of ATP or its non-hydrolysable analogue ATP-gamma-S for efficient interaction with other proteins including the protection of lambda O from aggregation. However, ClpX's ability to disaggregate lambda O aggregates requires hydrolysable ATP. We propose that the ClpX protein is a bona fide chaperone, whose biological role includes the maintenance of certain polypeptides in a form competent for proteolysis by the ClpP protease. Furthermore, our results suggest that the ClpX protein also performs typical chaperone protein functions independent of ClpP.

Isolation and characterization of ClpX, a new ATP-dependent specificity component of the Clp protease of Escherichia coli.

We have used 14C-labeled bacteriophage lambda O-DNA replication protein as a probe to identify and purify Escherichia coli proteases capable of its degradation. In this manner, five different proteases (termed Lop) have been identified capable of degrading lambda O protein to acid-soluble fragments in an ATP-dependent fashion. One of these activities was purified to homogeneity and shown to be composed of two different polypeptides. The 23,000-Da component (LopP) was identified as the previously characterized ClpP protein, known to complex with ClpA to form the ClpAP, an ATP-dependent protease, capable of degrading casein. The second 46,000-Da component was identified as ClpX (LopC), coded by a gene located in the same operon, but promoter distal to that coding for ClpP (Gottesman, S., Clark, W. P., de Crecy-Lagard, V., and Maurizi, M. R. (1993) J. Biol. Chem. 268, 22618-22626). This identification was based on the determination of the sequence of the first 24 amino acid residues of the purified ClpX protein and its identity with that predicted by the DNA sequence. The ClpXP protease is substrate specific, since it degrades casein (known to be degraded by ClpAP), lambda P, or DnaK proteins slowly or not at all. These results suggest that ClpX protein directs ClpP protease to specific substrates. It is estimated that 50% of all lambda O-specific protease activity present in crude E. coli extracts is due to the ClpXP protease. We propose that transient inhibition of lambda O degradation observed in vivo during the later stages of lambda-DNA replication in vivo is responsible for the switch from bidirectional to unidirectional replication. One round unidirectional replication will lead to strand separation resulting in a switch from early (theta) to late (sigma) mode of lambda-DNA replication.