Diss. Stockholm : Karolinska institutet, 2006.

Objective: Rare co-existance of disease or pathology Background: The lipid-rich variant is a rare and aggressive type of urothelial carcinoma (UCa), with less than 40 cases reported in the literature. This variant usually presents as an advanced-stage primary tumor. Case Report: We report the case of a 61-year-old man with previous history of T1 high-grade conventional urothelial carcinoma treated with local therapy. The patient later presented with a new 6.5-cm exophytic bladder mass. Histopathological examination revealed a T2 urothelial carcinoma of the lipid-rich variant. Retrospective review of the previous biopsies confirmed conventional high-grade urothelial car0cinoma, but scattered rare individual or small clusters of cells that resemble the lipid-rich variant urothelial carcinoma were also noted. Conclusions: The findings in this case suggest that the differential sensitivity of conventional urothelial carcinoma to local therapy may have allowed the lipid-rich variant to predominate in the recurrence. Pathologists should be aware of the lipid-rich variant of urothelial carcinoma. The prognostic significance of rare lipoblast-like cells among predominantly conventional urothelial carcinoma may requires further study.

京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第21255号 / 医博第4373号 / 新制||医||1029(附属図書館) / 京都大学大学院医学研究科医学専攻 / (主査)教授 長船 健二, 教授 柳田 素子, 教授 渡邊 直樹 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

Yes / Background: Many urothelial carcinomas (UC) contain activating PIK3CA mutations. In telomerase-immortalized normal urothelial cells (TERT-NHUC), ectopic expression of mutant PIK3CA induces PI3K pathway activation, cell proliferation and cell migration. However, it is not clear whether advanced UC tumors are PIK3CA-dependent and whether PI3K pathway inhibition is a good therapeutic option in such cases. Methods: We used retrovirus-mediated delivery of shRNA to knock down mutant PIK3CA in UC cell lines and assessed effects on pathway activation, cell proliferation, migration and tumorigenicity. The effect of the class I PI3K inhibitor GDC-0941 was assessed in a panel of UC cell lines with a range of known molecular alterations in the PI3K pathway. Results: Specific knockdown of PIK3CA inhibited proliferation, migration, anchorage-independent growth and in vivo tumor growth of cells with PIK3CA mutations. Sensitivity to GDC-0941 was dependent on hotspot PIK3CA mutation status. Cells with rare PIK3CA mutations and co-occurring TSC1 or PTEN mutations were less sensitive. Furthermore, downstream PI3K pathway alterations in TSC1 or PTEN or co-occurring AKT1 and RAS gene mutations were associated with GDC-0941 resistance. Conclusions: Mutant PIK3CA is a potent oncogenic driver in many UC cell lines and may represent a valuable therapeutic target in advanced bladder cancer.

Diss. (sammanfattning) Stockholm : Karol. inst., 2004. / Härtill 4 uppsatser.

Lactobacillus is a genus of lactic acid bacteria frequently used as healthpromoting probiotics. Using probiotics to treat or prevent infections is a novel experimental approach with vast impact on future therapy. Lactobacillus rhamnosus GR-1 is a probiotic investigated for its ability to reduce urogenital disease including urinary tract infections caused by pathogenic Escherichia coli. L. rhamnosus GR-1 has been shown to modulate immunity, thought to influence its probiotic effect. In this thesis, the aim was to study immunomodulation by L. rhamnosus GR-1 and other lactobacilli, with emphasis on elicited immune responses such as nuclear factor-kappaB (NF-κB) activation and cytokine release from human urothelial cells. Viable, heat-killed, and isolated released products from L. rhamnosus GR-1 augmented NF-κB activation in E. coli-challenged urothelial cells. Blocking of lipopolysaccharide binding to toll-like receptor 4 completely quelled this augmentation. Size-fractionation, urothelial cell challenge, and two-dimensional gel electrophoresis of L. rhamnosus GR-1 released products presented several candidate proteins with NF-κB modulatory actions including chaperonin GroEL, elongation factur Tu, and a protein from the NLP/P60 protein family. While tumor necrosis factor was correspondingly augmented by L. rhamnosus GR-1, the release of two other cytokines, interleukin (IL)-6 and CXCL8, was reduced. Similar effects were observed in macrophage-like cells stimulated with L. rhamnosus GR-1. Many immunomodulatory effects of lactobacilli are believed to be species and strain dependent. Therefore, twelve Lactobacillus strains were used to screen for their effects on CXCL8 release from urothelial cells. A majority of these strains were able to influence CXCL8 release from the cells. Phylogenetic analysis revealed close evolutionary linkage between lactobacilli with similar actions on CXCL8. Increased knowledge on probiotic bacterial products and the mechanism(s) of action could lead to improved future treatments for infections.

Orientador: Edson Antunes / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-23T00:32:19Z (GMT). No. of bitstreams: 1 Leiria_LuizOsorio_D.pdf: 1798358 bytes, checksum: ac871cdef8ebf1af12a2f5235c8d784d (MD5) Previous issue date: 2013 / Resumo: Obesidade/síndrome metabólica são fatores de risco para o desenvolvimento dos sintomas do trato urinário inferior (LUTS), incluindo hiperatividade de bexiga. Em nosso estudo visamos investigar a relação entre resistência à insulina e hiperatividade de bexiga em modelo de obesidade induzida por dieta hiperlipídica (10 semanas) em camundongos C57BL6/J. Curvas concentração-resposta a diferentes agentes contráteis e à insulina foram realizadas em bexigas isoladas de camundongos e humanos. Estudo cistométrico foi conduzido em camundongos anestesiados. Expressão protéica de PKC, canais de Ca2+ do tipo L, eNOS (Ser1177) e AKT (Ser473) fosforiladas, bem como de marcadores da resposta a proteínas mal dobradas (UPR), TRIB3, CHOP e ATF4, foi determinada por western blot. Camundongos obesos exibiram aumento do peso corpóreo, gordura epididimal, glicemia de jejum e resistência à insulina. As respostas contráteis aos diferentes agentes foram maiores nos animais obesos, o que foi normalizado pela pré-incubação com o bloqueador dos canais de Ca2+ do tipo L, amlodipino. Animais do grupo obeso apresentaram hiperatividade de bexiga, como demonstrado por estudo cistométrico. As curvas concentração-resposta ao agonista muscarínico, carbacol, foram dependentes da ativação da PKC. A resposta contrátil ao ativador da PKC, PDBu, foi abolida pela incubação com amlodipino. O tratamento com metformina melhorou a sensibilidade à insulina assim como normalizou a hipercontratilidade in vitro, a hiperatividade de bexiga e os níveis de PKC e pAKT em camundongos obesos. A insulina (1-100 nM) produziu relaxamento concentração-dependente em bexigas de camundongos e humanos, o que foi reduzido pela remoção da camada urotelial ou inibição da via de sinalização da PI3K/AKT/eNOS. Em condições fisiológicas, o estímulo com insulina resultou em um aumento de aproximadamente 3 vezes nos níveis de GMPc; porém, o relaxamento e os níveis de GMPc foram menores em bexigas de animais obesos. . A produção de cGMP foi reduzida na presença de inibidores da via da PI3K/AKT/eNOS. A inibição da PI3K aboliu a fosforilação de ambas eNOS e AKT no urotélio de camundongos. A expressão de pAKT e peNOS em resposta à insulina foi reduzida no urotélio de animais obesos em comparação com a do grupo controle, enquanto os níveis de TRIB3, CHOP e ATF4 foram maiores no urotélio dos animais obesos. O tratamento via oral com o inibidor de estresse de retículo endoplasmático, PBA, normalizou todos os parâmetros funcionais e moleculares dos camundongos obesos. Nossos dados mostram que a obesidade/síndrome metabólica em camundongos resulta em hiperatividade de bexiga associada à resistência local à insulina. A modulação positiva da PKC em camundongos obesos medeia simultaneamente à resistência à insulina em detrusor e a hiperatividade de bexiga. Além disso, camundongos obesos e resistentes à insulina exibem deficiência da ação da insulina na mucosa da bexiga como conseqüência de estresse de retículo, que parece estar também diretamente relacionado ao quadro de hiperatividade vesical nestes animais / Abstract: Obesity/metabolic syndrome are common risk factors for lower urinary tract symptoms, including overactive bladder. This study aimed to investigate whether insulin resistance affects bladder function in a model of obesity induced by high-fat diet (10 weeks) in C57BL6/J mice. Concentration-response curves to contractile agents and insulin were performed in human and mouse bladders. Cystometric study was performed in terminally anaesthetized mice. Western blot was performed in bladders to detect PKC, Cav1.2, phosphorylated eNOS (Ser1177) and AKT (Ser473), as well as the unfolded protein response (UPR) markers TRIB3, CHOP and ATF4. Obese mice exhibited higher body weight, epididymal fat mass, fasting glucose and insulin resistance. All the contractile agents produced greater bladder contractions in obese mice, which were fully reversed by the Cav1.2 blocker amlodipine. Cystometry evidenced overactive bladder in obese group that were also prevented by amlodipine. Carbachol induced contractions was dependent on the PKC activation, and PKC expression was increased in obese mice. Metformin treatment improved the insulin sensitivity, normalized the in vitro bladder hypercontractility, cystometric dysfunction and restored PKC and pAKT expression in the obese bladders. Insulin (1-100 nM) produced concentration-dependent mouse and human bladder relaxations that were markedly reduced by mucosal removal or inhibition of the PI3K/AKT/eNOS pathway. In mouse bladders, insulin produced a 3.0-fold increase in cGMP levels (P<0.05), that was prevented by PI3K/AKT/eNOS pathway inhibition. PI3K inhibition abolished insulin-induced phosphorylation of AKT and eNOS in bladder mucosa. Obese mice showed greater voiding frequency and non-voiding contractions, indicating overactive bladder. Insulin failed to relax bladder or to increase cGMP in obese group. Insulin-stimulated AKT and eNOS phosphorylation in mucosa was also impaired in obese mice. The UPR markers TRIB3, CHOP and ATF4 were increased in the mucosa of obese mice. The UPR inhibitor PBA normalized all the functional and molecular parameters in obese mice. Our data show that obesity/metabolic syndrome in mice resulted in overactive bladder associated to a local insulin resistance. Up-regulation of PKC in obese mice appears to mediate simultaneously the bladder overactivity and the insulin resistance in the bladders from obese mice bladder. Moreover, obese mice exhibited impaired insulin action in the urothelium layer as a consequence of the endoplasmic reticulum stress, which was demonstrated to be directly involved in the bladder overactivity of obese animals / Doutorado / Farmacologia / Doutor em Farmacologia

京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第20242号 / 医博第4201号 / 新制||医||1020(附属図書館) / 京都大学大学院医学研究科医学専攻 / (主査)教授 渡邊 直樹, 教授 岩井 一宏, 教授 楠見 明弘 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

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