The purpose of this study was to investigate several biological markers that may predict the response of muscle invasive transitional cell carcinoma TCC of the bladder to radical radiotherapy. The specific markers chosen were tumour angiogenesis (CD31 &CD34), tumour cell proliferation (Ki-67) and apoptosis (bcl-2), intratumour macrophage infiltration (CD68) and p53. Archival formalin fixed paraffin embedded pre treatment bladder biopsies from 101 patients with muscle invasive TCC were obtained. All patients subsequently received radical radiotherapy as their only treatment for the disease. 4µm sections were graded according to the WHO grading system and staged by the TNM classification. Angiogenesis (CD31&CD34) counts were obtained using a 25-point Chalkley eyepiece graticule. Bcl-2 scored as positive and negative, while p53, Ki-67 and CD68 were estimated using an eyepiece graticule. The medical records were examined to assess the response of the tumour at the 3-month post radiotherapy cystoscopy and the long-term outcome. Patients were classified into two groups in two sections: the first section includes (1) those free of disease (no tumour detected in the bladder at the 3-month cystoscopy), (2) those with resistant disease (tumour present at 3 months). The second section includes (1) those with persistent or recurrent cancer in the bladder (tumour recurred after an initial 3 months negative check cystoscopy together with patients with resistant disease at 3 months), (2) those free of disease at all subsequent cystoscopies. Detailed statistical analysis revealed that there were no association between any of the markers examined and the response to radiotherapy. MVD using CD34 was lower in higher stage tumours (p=0.050). Females, whilst representing only a small fraction (16) of the total of patients studied showed an inferior response to radiotherapy when compared to that of male patients (p=0.048). Higher median haemoglobin levels for the response group (p=0.031) was observed as well as a positive significant correlation between p53 (L1) expression and MIB-1 (LI) (r=0.332, p=0.001). The Kaplan-Meier survival analysis shows that the survival time is significantly better for those who were exposed longer to radiotherapy (> 33 days) (Log Rank, p=0.0246). There was a significantly higher survival time for patients who have CD68 higher than 42.4 (log rank, p=D.036). The study concluded that none of the selected markers could be used as prognostic value in determining patients most suitable for radiotherapy as primary treatment with curative intent for their bladder cancer. The finding of a poorer response in females is worthy of further study since, hormonal and anatomical influences may be important.

No description available.

No / The quinone based bioreductive drug apaziquone (EO9) failed to demonstrate efficacy in previous phase II studies following intravenous administration. We determined the dose of apaziquone that can be safely administered intravesically and explored its activity for superficial bladder transitional cell carcinoma. Six patients with multifocal, Ta/T1 and G1/G2 transitional cell carcinoma of the bladder received escalating doses of apaziquone formulated as EOquin¿ (0.5 mg/40 ml up to 16 mg/40 ml) weekly for 6 weeks. A further 6 patients received weekly apaziquone at the highest nontoxic dose established. Pharmacokinetic parameters were determined in urine and blood, and the pharmacodynamic markers NQO1 (reduced nicotinamide adenine dinucleotide phosphate:quinone oxidoreductase-1) and glucose transporter 1 were also characterized. Efficacy was determined against a marker lesion. Local toxicity (grades 2 and 3 dysuria, and hematuria) was observed at doses of 8 mg/40 ml and above but 4 mg/40 ml was well tolerated with no systemic or local side effects. Apaziquone in urine increased linearly with the dose but no apaziquone was detected in plasma. In 8 of 12 patients complete macroscopic and histological disappearance of the marker lesion occurred. A correlation between response and NQO1 and/or glucose transporter 1 expression could not be established. Intravesical administration of 4 mg/40 ml apaziquone was well tolerated and had ablative activity against superficial bladder cancer marker lesions.

No description available.

BACKGROUND: An important characteristic of transitional cell carcinoma (TCC) is the formation of tumors in multiple sites throughout the whole urinary tracts. Two theories explain the pathophysiologic mechanisms of multifocal tumors: (1) intraluminal seeding, it indicates the multiple tumors come from a single transformed malignant cell with secondary seeding or migration at different sites, and (2) the field cancerization, carcinogens affect the urothelium at multiple sites, leading to numerous mutation and independent growth of multifocal polyclonal tumors. Multifocal urothelial carcinomas could come from intraluminal seeding or from field cancerization. However, the data of clinical behaviors between the two different tumor types are lacking. METHODS: Bilateral synchronous and metachronous primary TCC of the upper urinary tracts were derived from field cancerization. Recurrent bladder cancers following upper-tract tumors mostly come from intraluminal seeding. The recurrence, progression, and prognosis of the two different tumors were analyzed. RESULTS: Bilateral upper-tract urothelial carcinomas derived from field cancerization were frequently associated with renal insufficiency, which were more invasive and had poor prognosis than bladder tumors derived from intraluminal seeding. CONCLUSION: The clinical behaviors of the multiple urothelial tumors derived from field cancerization and from intraluminal seeding should be different.

The purpose of this study is to investigate whether a prolonged delay in diagnosis of bladder cancer will result in worse outcomes for those patients, compared to those patients with a shorter diagnostic time interval. Data was collected on 247 patients newly diagnosed with transitional cell carcinoma of the bladder from January 1996 to December 2006 (10 years). The medical records of these patients were reviewed for demographics, pathological stage, date of consultation to the genitourinary (GU) service, and date of diagnosis by transurethral resection of bladder tumor (TURBT). The specialty delay was calculated as the time between the date of consultation to the GU service to the establishment of a diagnosis by TURBT. Univariate analyses were performed to test the association of specialty delay with clinical features and all-cause mortality. The median specialty delay in this study was 100 days. There was a trend towards a longer specialty delay for muscle-invasive disease (T2-T4) in comparison to superficial disease (Ta and T1). There was a significant correlation between all-cause mortality and increasing clinical stage (p=0.01). There was a paradoxical finding that patients with a specialty delay greater than 100 days had a significant reduction in all-cause death in comparison to patients with a specialty delay of 100 days or less (relative risk=0.59; 95% CI 0.36-0.90; p=0.01). In conclusion, this study did not confirm the hypothesis that a prolonged specialty delay in patients diagnosed with bladder cancer would result in a worse prognosis. In fact, there was a paradoxical finding that patients with a specialty delay greater than the median delay of 100 days had a better prognosis.

Made available in DSpace on 2014-06-11T19:33:25Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-02-16Bitstream added on 2014-06-13T20:24:56Z : No. of bitstreams: 1 silva_gn_dr_botfm.pdf: 930540 bytes, checksum: a962c4249ea11de03e0a230a9d385efb (MD5) / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Os agentes químicos utilizados para o tratamento do câncer ativam vias de sinalização que levam ao bloqueio ou retardo no ciclo celular, ativação ou desativação de mecanismos de reparo do DNA e apoptose. Entretanto, os mecanismos moleculares responsáveis por tais atividades ainda não foram completamente elucidados. Nesse contexto, os estudos de expressão gênica tornaram-se ferramentas importantes para a obtenção de informações que podem contribuir para o entendimento das respostas celulares frente à exposição a agentes neoplásicos e os mecanismos envolvidos na gênese e regressão de neoplasias. Em relação ao câncer de bexiga, a combinação de cisplatina/gencitabina é considerada como protocolo quimioterapêutico padrão. Entretanto, o mecanismo exato de ação dessas drogas ainda não foi completamente elucidado, principalmente no que diz respeito ao status do gene TP53, uma das principais alterações encontradas em câncer de bexiga. Com base nessas premissas, o objetivo deste estudo foi investigar os efeitos da cisplatina e da gencitabina sobre o índice apoptótico, ciclo celular e padrão de expressão gênica, em 3 linhagens de células de carcinoma transicional de bexiga (RT4, TP53 selvagem, grau 1; 5637, TP53 mutado, grau 2; T24, TP53 mutado, grau 3). Para avaliar tais respostas celulares foram utilizados os ensaios de citotoxicidade, sobrevivência celular e clonogência, análise do ciclo celular e apoptose por citometria de fluxo, e análise da expressão gênica usando microarranjos de cDNA. Os resultados mostraram: 1) que as células TP53 selvagem (RT4) foram mais sensíveis à apoptose do que células TP53 mutadas quando tratadas com cisplatina e gencitabina, ou com a combinação de ambas; 2) parada permanente do ciclo em G1, nas três linhagens celulares, após tratamento com a gencitabina e com a combinação cisplatina/gencitabina... / Background: currently, the combination cisplatin/gemcitabine is considered a (second) standard chemotherapeutical protocol for bladder cancer. However, the mechanism by which these drugs act on tumor cells is not completely understood. Therefore, the aim of the present was to investigate the effects of these two antineoplastic drugs on the apoptotic index and cell cycle distribution in urinary bladder transitional cell carcinoma (TCC) cell lines with different TP53 status (RT4, with wild type TP53; 5637 and T24, with mutated TP53). Methods: cytotoxicity, cell survival and clonogenic survival assays, and flow citometry analysis for cell cycle and apoptosis cells were done in the cell lines treated with different concentrations of cisplatin and gemcitabine. Results: wild type TP53 cells were more sensitive to apoptosis than mutated TP53 cells when treated with cisplatin or gemcitabine; permanent G1 cell-cycle arrest was observed in the three cell lines after treatment with gemcitabine or with the combination of both; significant cell death was detected in all the cell lines after treatments with cisplatin or gemcitabine. However, when drugs were given in combinations, lower percentage of survival occurred independent on TP53 status. Conclusions: it was demonstrated that the combination of cisplatin and gemcitabine can be effective despite of tumor-associated TP53 mutations. Thus, the synergism between low concentrations of cisplatin and gemcitabine may have clinical relevance, since the use of high concentrations of cisplatin is toxic to the whole organism.

Orientador: Daisy Maria Fávero Salvadori / Banca: Maria Inês de Moura Campos Pardini / Banca: Katia Ramos Moreira Leite / Banca: Daniel Araki Ribeiro / Banca: Paulo Peitl Júnior / Resumo: Os agentes químicos utilizados para o tratamento do câncer ativam vias de sinalização que levam ao bloqueio ou retardo no ciclo celular, ativação ou desativação de mecanismos de reparo do DNA e apoptose. Entretanto, os mecanismos moleculares responsáveis por tais atividades ainda não foram completamente elucidados. Nesse contexto, os estudos de expressão gênica tornaram-se ferramentas importantes para a obtenção de informações que podem contribuir para o entendimento das respostas celulares frente à exposição a agentes neoplásicos e os mecanismos envolvidos na gênese e regressão de neoplasias. Em relação ao câncer de bexiga, a combinação de cisplatina/gencitabina é considerada como protocolo quimioterapêutico padrão. Entretanto, o mecanismo exato de ação dessas drogas ainda não foi completamente elucidado, principalmente no que diz respeito ao status do gene TP53, uma das principais alterações encontradas em câncer de bexiga. Com base nessas premissas, o objetivo deste estudo foi investigar os efeitos da cisplatina e da gencitabina sobre o índice apoptótico, ciclo celular e padrão de expressão gênica, em 3 linhagens de células de carcinoma transicional de bexiga (RT4, TP53 selvagem, grau 1; 5637, TP53 mutado, grau 2; T24, TP53 mutado, grau 3). Para avaliar tais respostas celulares foram utilizados os ensaios de citotoxicidade, sobrevivência celular e clonogência, análise do ciclo celular e apoptose por citometria de fluxo, e análise da expressão gênica usando microarranjos de cDNA. Os resultados mostraram: 1) que as células TP53 selvagem (RT4) foram mais sensíveis à apoptose do que células TP53 mutadas quando tratadas com cisplatina e gencitabina, ou com a combinação de ambas; 2) parada permanente do ciclo em G1, nas três linhagens celulares, após tratamento com a gencitabina e com a combinação cisplatina/gencitabina... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Background: currently, the combination cisplatin/gemcitabine is considered a (second) standard chemotherapeutical protocol for bladder cancer. However, the mechanism by which these drugs act on tumor cells is not completely understood. Therefore, the aim of the present was to investigate the effects of these two antineoplastic drugs on the apoptotic index and cell cycle distribution in urinary bladder transitional cell carcinoma (TCC) cell lines with different TP53 status (RT4, with wild type TP53; 5637 and T24, with mutated TP53). Methods: cytotoxicity, cell survival and clonogenic survival assays, and flow citometry analysis for cell cycle and apoptosis cells were done in the cell lines treated with different concentrations of cisplatin and gemcitabine. Results: wild type TP53 cells were more sensitive to apoptosis than mutated TP53 cells when treated with cisplatin or gemcitabine; permanent G1 cell-cycle arrest was observed in the three cell lines after treatment with gemcitabine or with the combination of both; significant cell death was detected in all the cell lines after treatments with cisplatin or gemcitabine. However, when drugs were given in combinations, lower percentage of survival occurred independent on TP53 status. Conclusions: it was demonstrated that the combination of cisplatin and gemcitabine can be effective despite of tumor-associated TP53 mutations. Thus, the synergism between low concentrations of cisplatin and gemcitabine may have clinical relevance, since the use of high concentrations of cisplatin is toxic to the whole organism. / Doutor

No description available.

*1-Dimensional Advective-Diffusion Model in Porous Media Infusion of therapeutic agents into tissue is makes use of two mass transport modes: advective transport, and molecular diffusion. Bulk infusion into a 0.6% wt agarose phantom was modeled as an infinite, homogenous, and isotropic porous medium saturated with the same solvent used in the infused dye tracer. The source is assumed to be spherical and isotropic with constant flow rate and concentration. The Peclet numberdecreases with power function Pe = 15762t0.337 due to the decrease in mean dye-front pore velocity as V goes to Vfinal. Diffusive mass transport does not become significant during any relevent time period. *Arborizing Fiberoptic Microneedle Catheter We have developed an arborizing catheter that allows multiple slender fused-silica CED cannulae to be deployed within a target volume of the brain via a single needle tract, and tested it in a widely accepted tissue phantom. The arborizing catheter was constructed by bonding and encapsulating seven slender PEEK tubes in a radially symmetric bundle with a progressive helical angle along the length, then grinding a conicle tip where the helical angle is greatest. The catheter was tested by casting 0.6% wt agarose around the device with all needles deployed to a tip-to-tip distance of 4 mm. Phantom temperature was maintained at 26 ± 2°C. 5% wt Indigo Carmine dye was infused at a rate of 0.3 uL/min/needle for 4 hours. N=4 infusions showed a Vd/Vi of 139.774, with a standard deviation of 45.01. This is an order of magnitude greater than single-needle infusions under similar conditions [45]. The arborizer showed the additional benefit of arresting reflux propagating up the lengths of individual needles, which has historically been a weakness of single-needle CED catheter designs. *In Vivo Co-Delivery of Single Walled Carbon Nano-horns and Laser Light to Treat Human Transitional Cell Carcinoma of the Urinary Bladder in a Rodent Model Using a rodent model we explored a treatment method for Transitional Cell Carcinoma (TCC) in the urinary bladder in which Single Walled Carbon Nanohorn (SWNH) solution and 1064 nm laser light are delivered into tumorous tissue via a co-delivery Fiberoptic Microneedle Device (FMD). Preliminary treatment parameters were determined by injecting SWNH solutions with concentrations of 0 mg/mL, 0.17 mg/mL, or 0.255 mg/mL into ex vivo porcine skin and irradiating each for three minutes at laser powers of 500 mW, or 1000 mW. The combination with the greatest temperature increase without burning the tissue, 0.17 mg/mL at 1000 mW, was selected for the in vivo treatment. TCC tumors were induced in a rodent model by injecting a solution of 106 AY27 urothelial carcinoma cells into the lateral aspect of the left hind leg of young, female F344 rats. When tumors reached 5-10 mm3, rats were anesthitized and treated. SWNH solution was injected directly into the tumor and irradiated until the target temperature of 60degC was achieved. The rats were then recovered from anestesia and monitored for 7-14 days, at which point they were humanely sacrificed, and the tumors prepared for histological examination. Histological assessment of areas of FMD treatment correlated well with gross morphological appearance. Foci of tumor necrosis showed sharp (1-2 mm) delineation from areas of viable tumor (not treated) and normal tissue. We believe we have demonstrated the feasibility of using the FMD for treatment of urothelial carcinoma using an animal model of this disease, and are encouraged to continue development of this treatment and testing in larger animal models. / Master of Science