RESUMO
Bladder cancer, one of the most prevalent malignancies worldwide, remains hard to classify due to a staggering molecular complexity. Despite a plethora of diagnostic tools and therapies, it is hard to outline the key steps leading up to the transition from high-risk non-muscle-invasive bladder cancer (NMIBC) to muscle-invasive bladder cancer (MIBC). Carcinogen-induced murine models can recapitulate urothelial carcinogenesis and natural anti-tumor immunity. Herein, we have developed and profiled a novel model of progressive NMIBC based on 10 weeks of OH-BBN exposure in hepatocyte growth factor/cyclin dependent kinase 4 (R24C) (Hgf-Cdk4R24C) mice. The profiling of the model was performed by histology grading, single cell transcriptomic and proteomic analysis, while the derivation of a tumorigenic cell line was validated and used to assess in vivo anti-tumor effects in response to immunotherapy. Established NMIBC was present in females at 10 weeks post OH-BBN exposure while neoplasia was not as advanced in male mice, however all mice progressed to MIBC. Single cell RNA sequencing analysis revealed an intratumoral heterogeneity also described in the human disease trajectory. Moreover, although immune activation biomarkers were elevated in urine during carcinogen exposure, anti-programmed cell death protein 1 (anti-PD1) monotherapy did not prevent tumor progression. Furthermore, anti-PD1 immunotherapy did not control the growth of subcutaneous tumors formed by the newly derived urothelial cancer cell line. However, treatment with CpG-oligodeoxynucleotides (ODN) significantly decreased tumor volume, but only in females. In conclusion, the molecular map of this novel preclinical model of bladder cancer provides an opportunity to further investigate pharmacological therapies ahead with regards to both targeted drugs and immunotherapies to improve the strategies of how we should tackle the heterogeneous tumor microenvironment in urothelial bladder cancer to improve responses rates in the clinic.
Assuntos
Cálculos da Bexiga Urinária/metabolismo , Animais , Butilidroxibutilnitrosamina/farmacologia , Carcinógenos/farmacologia , Linhagem Celular Tumoral , Modelos Animais de Doenças , Feminino , Humanos , Estudos Longitudinais , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Reação em Cadeia da Polimerase , Proteinúria/urina , Proteômica/métodos , Análise de Sequência de RNA , Análise de Célula Única , Bexiga Urinária/efeitos dos fármacos , Bexiga Urinária/metabolismo , Bexiga Urinária/patologia , Cálculos da Bexiga Urinária/induzido quimicamente , Cálculos da Bexiga Urinária/urina , Urotélio/efeitos dos fármacos , Urotélio/metabolismo , Urotélio/patologiaRESUMO
AIM: To assess the high (18)F-fluorodeoxyglucose ((18)F-FDG) uptake in urinary calculi on positron-emission tomography/computed tomography (PET/CT). METHODS: In this study, (18)F-FDG PET/CT examinations were retrospectively reviewed from November 2013 to February 2016 in a single center, and patients with high (18)F-FDG uptake in urinary calculi were identified. The following data were collected from each patient, including age, sex, primary disease, method to verify the urinary calculus, and imaging characteristics of the calculus. RESULTS: A total of 2758 PET/CT studies (2567 patients) were reviewed, and 52 patients with urinary calculi were identified, in which 6 (11.5%, 6/52) patients (5 males, 1 female, age 34-73 years, median age 60.5 years) demonstrated high (18)F-FDG uptake in the urinary calculi. Among the 6 patients, 3 patients had bladder calculi, 2 patients had renal calculi, and 1 patient had both bladder and renal calculi. The size of the urinary calculi varied from sandy to 19mm on CT. The maximal Hounsfield units of the calculi ranged from 153 to 1078. The SUVmax of the calculi on the routine PET/CT scan ranged from 11.7 to 143.0. Delayed PET/CT scans were performed on 4 patients, which showed the calculi SUVmax increasing in 2 patients, while decreasing in the other 2 patients. One patient with bladder calculus underwent a follow-up PET/CT, which showed enlargement of the calculus as well as the increased SUVmax. CONCLUSION: This study shows an uncommon high (18)F-FDG uptake in urinary calculi. Recognition of this non-malignant accumulation in urinary calculi is essential for correct interpretation of PET/CT findings.
Assuntos
Fluordesoxiglucose F18/farmacocinética , Tomografia por Emissão de Pósitrons combinada à Tomografia Computadorizada/métodos , Compostos Radiofarmacêuticos/farmacocinética , Cálculos Urinários/metabolismo , Adulto , Idoso , Feminino , Seguimentos , Humanos , Aumento da Imagem/métodos , Cálculos Renais/diagnóstico por imagem , Cálculos Renais/metabolismo , Masculino , Pessoa de Meia-Idade , Intensificação de Imagem Radiográfica/métodos , Estudos Retrospectivos , Cálculos da Bexiga Urinária/diagnóstico por imagem , Cálculos da Bexiga Urinária/metabolismo , Cálculos Urinários/diagnóstico por imagemRESUMO
The catheter-associated uropathogenProteus mirabilisfrequently causes urinary stones, but little has been known about the initial stages of bladder colonization and stone formation. We found thatP. mirabilisrapidly invades the bladder urothelium, but generally fails to establish an intracellular niche. Instead, it forms extracellular clusters in the bladder lumen, which form foci of mineral deposition consistent with development of urinary stones. These clusters elicit a robust neutrophil response, and we present evidence of neutrophil extracellular trap generation during experimental urinary tract infection. We identified two virulence factors required for cluster development: urease, which is required for urolithiasis, and mannose-resistantProteus-like fimbriae. The extracellular cluster formation byP. mirabilisstands in direct contrast to uropathogenicEscherichia coli, which readily formed intracellular bacterial communities but not luminal clusters or urinary stones. We propose that extracellular clusters are a key mechanism ofP. mirabilissurvival and virulence in the bladder.
Assuntos
Proteínas de Bactérias , Fímbrias Bacterianas , Infecções por Proteus , Proteus mirabilis , Urease , Cálculos da Bexiga Urinária , Animais , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Modelos Animais de Doenças , Feminino , Fímbrias Bacterianas/genética , Fímbrias Bacterianas/metabolismo , Camundongos , Camundongos Endogâmicos CBA , Infecções por Proteus/genética , Infecções por Proteus/metabolismo , Infecções por Proteus/patologia , Proteus mirabilis/genética , Proteus mirabilis/metabolismo , Proteus mirabilis/patogenicidade , Urease/genética , Urease/metabolismo , Bexiga Urinária/microbiologia , Bexiga Urinária/patologia , Cálculos da Bexiga Urinária/genética , Cálculos da Bexiga Urinária/metabolismo , Cálculos da Bexiga Urinária/microbiologia , Cálculos da Bexiga Urinária/patologia , Escherichia coli Uropatogênica/genética , Escherichia coli Uropatogênica/metabolismo , Escherichia coli Uropatogênica/patogenicidadeRESUMO
We present here two special cases of urolithiasis. The first one shows a giant bladder lithiasis resulting in severe renal insufficiency in a 63-year-old patient, who had previously had nicturia (2-3 times), occasional episodes of urinary frequency and burning micturition, in the absence of renal colic, hematuria or interrupted urination. The second case referes to an 85-year-old man suffering from prostatic enlargement and bladder stones, hospitalized to undergo intervention of trans-vesical prostatic adenomectomy, during which two star-shaped stones were found without obvious symptoms.
Assuntos
Cálculos da Bexiga Urinária/patologia , Idoso de 80 Anos ou mais , Carbonatos/análise , Humanos , Masculino , Pessoa de Meia-Idade , Oxalatos/análise , Fosfatos/análise , Hiperplasia Prostática/complicações , Hiperplasia Prostática/cirurgia , Infecções por Pseudomonas/complicações , Ácido Úrico/análise , Cálculos da Bexiga Urinária/química , Cálculos da Bexiga Urinária/complicações , Cálculos da Bexiga Urinária/metabolismo , Cálculos da Bexiga Urinária/cirurgia , Infecções Urinárias/complicações , Infecções Urinárias/microbiologiaRESUMO
OBJECTIVE: To evaluate the mechanisms of bladder uric acid stone (BUAS) formation by analyzing BUAS stone matrix proteins, with mass spectrometry (MS). MATERIALS AND METHODS: Stone matrix proteins were extracted from 5 pure BUASs. The obtained proteins were analyzed with reverse phase liquid chromatography-tandem MS. The acquired data were investigated against a Swiss Prot human protein database, using Matrix Science Mascot. The identified proteins were submitted to UniProtKB website for gene ontology analysis to define their correlation. They were also submitted to Metacore platform and Kyoto Encyclopedia of Genes and Genomes website for pathway analysis. MS-determined protein expressions were validated by immunoblot. RESULTS: The liquid chromatography-tandem MS analysis identified 58-226 proteins in the 5 BUASs (450 proteins). Metacore software analysis suggests that inflammation might play an important role for BUAS formation. The analysis of endogenous metabolic pathways revealed that these proteins were categorized into glycerophospholipid or glycosphingolipid biosynthesis. Four of 5 identified proteins selected for validation, including uromodulin, S100P, Histone 4, and nucleophosmin, can be validated in the immunoblot data. CONCLUSION: Our results suggest that inflammatory process and lipid metabolism might play a role in the formation of BUAS. Whether these inflammatory responses are the etiology of stone formation or whether they result from local damage by stone irritation is uncertain.
Assuntos
Proteínas/análise , Cálculos da Bexiga Urinária/química , Cálculos da Bexiga Urinária/metabolismo , Vias Biossintéticas , Proteínas de Ligação ao Cálcio/análise , Cromatografia Líquida , Cistite/complicações , Cistite/metabolismo , Glicerofosfolipídeos/biossíntese , Glicoesfingolipídeos/biossíntese , Humanos , Metabolismo dos Lipídeos , Proteínas de Neoplasias/análise , Mapeamento de Peptídeos , Proteínas/metabolismo , Espectrometria de Massas em Tandem , Ácido Úrico , Cálculos da Bexiga Urinária/etiologia , Uromodulina/análiseRESUMO
Melamine can cause urinary stones related to nephropathy of the kidney and hyperplasia or carcinoma of the bladder, but the mechanism of stone formation is not well understood. In this study, male rats were administered melamine for thirteen weeks to establish melamine bladder stone models and the stones were analysed by Fourier transform infrared (FTIR) spectroscopy, powder X-ray diffraction (XRD), energy dispersive X-ray (EDX) spectroscopy, sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE), liquid chromatography/mass spectrometry/mass spectrometry (LC-MS/MS) and western blot, respectively, for the composition and proteome, and to explore the implication of proteins for stone formation. The results showed bladder stones were composed of predominant melamine and a few amount of proteins. The proteins had a wide range of molecular weights and 1051 proteins were identified. Gene Ontology (GO) classification of the identified proteins showed most proteins were from injured cells, involved in various metabolic processes and had binding functions. Of the identified proteins, there were a few inflammatory proteins and urinary proteins. Physicochemical characteristics of the identified proteins showed that 67.1% proteins' isoelectric points (pI) value was below 7.0, 91.1% proteins' grand average of hydropathicity (GRAVY) scores were below 0 and nearly half of the proteins were stable. Our data indicated proteins might play an important role in melamine bladder stone formation.
Assuntos
Modelos Animais de Doenças , Proteoma/efeitos dos fármacos , Resinas Sintéticas/toxicidade , Triazinas/toxicidade , Cálculos da Bexiga Urinária/induzido quimicamente , Animais , Western Blotting , Cromatografia Líquida de Alta Pressão , Eletroforese em Gel de Poliacrilamida , Masculino , Proteoma/química , Proteoma/metabolismo , Ratos , Ratos Sprague-Dawley , Resinas Sintéticas/química , Resinas Sintéticas/metabolismo , Espectrometria por Raios X , Espectroscopia de Infravermelho com Transformada de Fourier , Espectrometria de Massas em Tandem , Triazinas/química , Triazinas/metabolismo , Cálculos da Bexiga Urinária/química , Cálculos da Bexiga Urinária/metabolismo , Difração de Raios XRESUMO
Mitochondria accumulate within nerve terminals and support synaptic function, most notably through ATP production. They can also sequester Ca(2+) during nerve stimulation, but it is unknown whether this limits presynaptic Ca(2+) levels at physiological nerve firing rates. Similarly, it is unclear whether mitochondrial Ca(2+) sequestration differs between functionally different nerve terminals. We addressed these questions using a combination of synthetic and genetically encoded Ca(2+) indicators to examine cytosolic and mitochondrial Ca(2+) levels in presynaptic terminals of tonic (MN13-Ib) and phasic (MNSNb/d-Is) motor neurons in Drosophila, which, as we determined, fire during fictive locomotion at approximately 42 Hz and approximately 8 Hz, respectively. Mitochondrial Ca(2+) sequestration starts in both terminals at approximately 250 nM, exhibits a similar Ca(2+)-uptake affinity (approximately 410 nM), and does not require Ca(2+) release from the endoplasmic reticulum. Nonetheless, mitochondrial Ca(2+) uptake in type Is terminals is more responsive to low-frequency nerve stimulation and this is due to higher cytosolic Ca(2+) levels. Since type Ib terminals have a higher mitochondrial density than Is terminals, it seemed possible that greater mitochondrial Ca(2+) sequestration may be responsible for the lower cytosolic Ca(2+) levels in Ib terminals. However, genetic and pharmacological manipulations of mitochondrial Ca(2+) uptake did not significantly alter nerve-stimulated elevations in cytosolic Ca(2+) levels in either terminal type within physiologically relevant rates of stimulation. Our findings indicate that presynaptic mitochondria have a similar affinity for Ca(2+) in functionally different nerve terminals, but do not limit cytosolic Ca(2+) levels within the range of motor neuron firing rates in situ.
Assuntos
Cálcio/metabolismo , Mitocôndrias/metabolismo , Neurônios Motores/metabolismo , Terminações Pré-Sinápticas/metabolismo , Animais , Drosophila , Larva , Mitocôndrias/ultraestrutura , Neurônios Motores/ultraestrutura , Terminações Pré-Sinápticas/ultraestrutura , Cálculos da Bexiga Urinária/metabolismoRESUMO
BACKGROUND: The effects of bisphosphonates on prophylaxis of stone formation are unclear. We evaluated the outcome of two new-generation bisphosphonates in a lithogenic rat model. METHODS: 36 male rats were divided into three groups of 12 animals each. Both calcium and creatinine levels of plasma and urine were measured. Zinc discs of about 40 mg each were surgically placed into the bladder. The first group received no treatment and the second and third groups were treated with an intraperitoneal injection of weekly clodronate (20 mg/kg) and zoledronic acid (7.5 microg/kg), respectively. At the end of the 8th week, the weight increase in discs and biochemical changes were analyzed comparatively. RESULTS: The mean weight of discs in the control, clodronate and zoledronic acid groups was 109.65 +/- 80.97, 79.82 +/- 17.99 and 72.91 +/- 19.29 mg, respectively (p > 0.05). The percentage increase in weight of discs was 164% for control, 90% for clodronate and 71% for the zoledronic acid group. The increase of urinary calcium level in the zoledronic acid group was lower than the others (p < 0.05). CONCLUSIONS: When considering the percentage increase in weight of discs, the difference between control and bisphosphonate groups support the idea that these drugs may have a preventive role in stone formation.
Assuntos
Oxalato de Cálcio/metabolismo , Ácido Clodrônico/farmacologia , Difosfonatos/farmacologia , Imidazóis/farmacologia , Cálculos da Bexiga Urinária/prevenção & controle , Animais , Cálcio/sangue , Cálcio/urina , Ácido Clodrônico/administração & dosagem , Creatinina/sangue , Creatinina/urina , Difosfonatos/administração & dosagem , Modelos Animais de Doenças , Imidazóis/administração & dosagem , Injeções Intraperitoneais , Masculino , Ratos , Ratos Sprague-Dawley , Cálculos da Bexiga Urinária/induzido quimicamente , Cálculos da Bexiga Urinária/metabolismo , Zinco , Ácido ZoledrônicoRESUMO
Prolonged cell proliferation in response to irritation by bladder calculi can evoke malignant transformation of the urothelium. However, the molecular mechanisms responsible for calculi-associated bladder carcinogenesis are unknown. We compared the protein expression pattern of rat bladder transitional cell carcinomas (TCCs) induced by terephthalic acid with that of normal bladder tissues using 2-DE. Comparative analysis of the respective spot patterns on 2-DE showed 146 spots that were markedly changed in TCC samples. Subsequently, 56 of the variant protein spots were identified by MALDI-TOF MS. Among them, overexpression of annexin a1 (ANNA1) in rat TCCs was confirmed by Western blotting and real-time RT-PCR analysis. Immunohistochemical staining revealed that ANNA1, usually a cytoplasmic protein in normal urothelium, was translocated to the nucleus in rat bladder cancer cells. In contrast to the animal studies, examination of human clinical specimens showed that ANNA1 expression was reduced in TCC compared to normal urothelium. The expression of ANNA1 was inversely related to the level of differentiation of TCC. Our data suggest that overexpression of ANNA1 is involved in bladder carcinogenesis induced by bladder calculi and that translocation of the protein may be partly responsible for the effect. ANNA1 may serve as a new marker of differentiation for the histopathological grading of human TCC.
Assuntos
Anexina A1/genética , Carcinoma de Células de Transição/genética , Regulação Neoplásica da Expressão Gênica/genética , Neoplasias da Bexiga Urinária/genética , Adulto , Idoso , Idoso de 80 Anos ou mais , Animais , Anexina A1/biossíntese , Anexina A1/química , Western Blotting , Carcinoma de Células de Transição/induzido quimicamente , Carcinoma de Células de Transição/metabolismo , Diferenciação Celular/efeitos dos fármacos , Núcleo Celular/metabolismo , Modelos Animais de Doenças , Eletroforese em Gel Bidimensional/métodos , Feminino , Humanos , Imuno-Histoquímica , Masculino , Pessoa de Meia-Idade , Ácidos Ftálicos , Ratos , Ratos Wistar , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Cálculos da Bexiga Urinária/induzido quimicamente , Cálculos da Bexiga Urinária/genética , Cálculos da Bexiga Urinária/metabolismo , Neoplasias da Bexiga Urinária/induzido quimicamente , Neoplasias da Bexiga Urinária/metabolismoRESUMO
Nongenotoxic bladder carcinogens that form bladder calculi have been concluded to be of low carcinogenic risk to humans because bladder stones would be expelled or surgically removed before they had a chance to exert their carcinogenic effect. It is the aim of this report to examine the possible contribution of indomethacin to the carcinogenic risk posed by nongenotoxic bladder carcinogens that cause bladder stones. Indomethacin may act as a tumor promoter in the bladder by interfering with the synthesis of prostaglandins. Prostaglandins have a cytoprotective function in the gastric mucosa and possibly also in the urinary bladder. Diminished cytoprotection may be implicated in bladder carcinogenesis as beta-naphthylamine, a human bladder carcinogen, also inhibits prostaglandin synthesis in vitro. The presence of other tumor promoters in the bladder may further ensure that tumors would be formed even if bladder stones were expelled. People who are exposed to nongenotoxic bladder carcinogens that are present in the environment and that form bladder stones, therefore, may be at an increased risk for developing bladder cancer if they are also exposed to tumor promoters, such as indomethacin.
Assuntos
Carcinógenos/toxicidade , Transformação Celular Neoplásica/efeitos dos fármacos , Cocarcinogênese , Inibidores de Ciclo-Oxigenase/toxicidade , Indometacina/toxicidade , Cálculos da Bexiga Urinária/induzido quimicamente , Neoplasias da Bexiga Urinária/etiologia , Animais , Transformação Celular Neoplásica/metabolismo , Humanos , Prostaglandinas/metabolismo , Medição de Risco , Cálculos da Bexiga Urinária/complicações , Cálculos da Bexiga Urinária/metabolismo , Neoplasias da Bexiga Urinária/metabolismoRESUMO
To investigate the prostaglandin E2 (PGE2) biosynthetic mechanism in bladder carcinogenesis, we established Wistar rat model of bladder papilloma and transitional cell carcinoma (TCC) induced by 5% terephthalic acid (TPA) treatment. Then, the mRNA level of cytosolic phospholipase A2 (cPLA2), cyclooxygenases (COX)-1 and -2, membrane-bound PGE2 synthases (mPGES)-1 and -2 was detected using reverse transcription polymerase chain reaction (RT-PCR). Immunoblotting was applied to detect the expression of COX-2 protein. Proliferating cell nuclear antigen (PCNA) was determined by immunohistochemistry. In addition, the level of PGE2 was measured by radioimmunoassay (RIA). Bladder papilloma (100%, 8/8) was examined in rats after 24-week treatment, and bladder TCC (80%, 16/20) was found after 48-week treatment. Histopathological changes were not found in control group rats. The incidence of bladder papilloma and TCC in test group was significantly higher than that in control group (P<0.01). The mRNA levels of cPLA2, COX-2 and mPGES-1 in the bladder papilloma and TCC were significantly higher than those in normal bladder (P<0.01), while the mRNA levels of COX-1 and mPGES-2 in TCC were unchanged compared with normal bladder. Bladder TCC exhibited a substantial expression of COX-2 protein. On the contrary, normal bladder tissue barely expresses COX-2 protein. PCNA labeling index (LI) and the level of PGE2 in bladder papilloma are much higher than those in normal bladder (P<0.01), but lower than those in bladder TCC (P<0.05). In conclusion, increasing PGE2 level via cPLA2--COX-2--mPGES-1 pathway may play an important role in rat bladder carcinogenesis. PGE2 may be a biomarker for the development of bladder TCC. cPLA2 and mPGES-1 may be targets for development of novel chemoprevention strategies for bladder TCC.
Assuntos
Ciclo-Oxigenase 2/metabolismo , Dinoprostona/metabolismo , Oxirredutases Intramoleculares/metabolismo , Fosfolipases A/metabolismo , Ácidos Ftálicos/toxicidade , Neoplasias da Bexiga Urinária/metabolismo , Animais , Sequência de Bases , Western Blotting , Carcinoma de Células de Transição/induzido quimicamente , Carcinoma de Células de Transição/genética , Carcinoma de Células de Transição/metabolismo , Transformação Celular Neoplásica/efeitos dos fármacos , Transformação Celular Neoplásica/genética , Transformação Celular Neoplásica/metabolismo , Ciclo-Oxigenase 2/genética , Fosfolipases A2 do Grupo IV , Imuno-Histoquímica , Oxirredutases Intramoleculares/genética , Isoenzimas/genética , Isoenzimas/metabolismo , Masculino , Dados de Sequência Molecular , Papiloma/induzido quimicamente , Papiloma/genética , Papiloma/metabolismo , Fosfolipases A/genética , Fosfolipases A2 , Prostaglandina-E Sintases , Distribuição Aleatória , Ratos , Ratos Wistar , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Cálculos da Bexiga Urinária/induzido quimicamente , Cálculos da Bexiga Urinária/genética , Cálculos da Bexiga Urinária/metabolismo , Neoplasias da Bexiga Urinária/induzido quimicamente , Neoplasias da Bexiga Urinária/genéticaRESUMO
Long-term histopathologic changes after bladder augmentation (BA) in rats using living-related partial bladder transplantation (LPBTx) or conventional ileocystoplasty (ICP) were compared. In this study, BA (n = 37), LPBTx (n = 18), and ICP (n = 19) were performed in 16-wk-old Lewis rats. Five donors and seven nontransplanted normal Lewis rats (controls) were also studied. Rats that survived >10 mo after BA were killed after blood biochemistry and neobladder imaging. Harvested bladders were examined with hematoxylin and eosin and proliferating cell nuclear antigen (PCNA). When the rats were killed, there were 16 rats in the LPBTx group and 12 rats in the ICP group; ICP rats were significantly smaller than LPBTx rats (p < 0.05). Mean duration of follow-up for the LPBTX group was 17.3 mo, for the ICP group was 13.7 mo, for the donor group was 16.1 mo, and for the control group was 19.7 mo. Mean serum pH in the LPBTx group was 7.41 +/- 0.78 and in the ICP group was 7.25 +/- 0.38. Mean base excess in the ICP group was significantly lower than in the LPBTx group (p < 0.05). Incidence of bladder calculi in the LPBTx group (6.3%) was significantly lower than in the ICP group (33.3%; p < 0.05). There was no dysplasia/malignancy/increase in PCNA in the LPBTx group. PCNA increased in the ICP group, compared with controls (p < 0.05); two (16.7%) of 12 of ICP rats had dysplasia with mitosis. Bladder capacity increased in LPBTx and ICP compared with controls (both p < 0.05). We hope to show that BA using LPBTx may result in a neobladder with fewer complications than BA using ICP; LPBTx may also decrease the risk for malignancy.
Assuntos
Transplante Heterotópico/métodos , Bexiga Urinária/cirurgia , Procedimentos Cirúrgicos Urológicos/métodos , Anastomose Cirúrgica/métodos , Animais , Feminino , Modelos Animais , Antígeno Nuclear de Célula em Proliferação/metabolismo , Ratos , Ratos Endogâmicos Lew , Fatores de Tempo , Coleta de Tecidos e Órgãos/métodos , Transplante Isogênico , Resultado do Tratamento , Bexiga Urinária/patologia , Cálculos da Bexiga Urinária/metabolismoRESUMO
To obtain definitive information about the mechanisms of urinary calculus formation and the structural characteristics of the calculi induced by biphenyl administration in rats, with a focus on the sex dependency, the constituents of the urinary calculi were analyzed by HPLC, inductively coupled plasma spectroscopy (ICP), micro Fourier transform infrared spectroscopy (mFT-IR), and ion chromatography (IC), and structural analyses were carried out by microscopy, mFT-IR, and the electron probe microanalyzer (EPMA) method. We attempted to account for the appreciably higher incidence of calculi in males than in females. mFT-IR analysis revealed that the biphenyl-induced urinary calculi in male rats are composed mainly of potassium 4-hydroxybiphenyl-o-sulfate (4-HBPOSK), whereas the calculi in female rats are composed mainly of 4-hydroxybiphenyl (4-HBP) and KHSO(4) produced by the hydrolysis of 4-HBPOSK. Observations of photomicrographs and the results of mFT-IR analysis indicated that the calculi in males have a multilayer structure consisting of alternating layers of 4-HBPOSK and calcium phosphate, whereas the calculi in females have no multilayer structure, but open holes in which needle-shaped crystals are present in some places. In view of the results of these analyses, including the EPMA analysis, it appears that calculus formation in males may involve a series of successive and irreversible reactions, whereas calculus formation in females may result from a series of reversible reactions, including the hydrolysis of 4-HBPOSK. It was inferred that the series of irreversible reactions involved in calculus formation in males is relatively more stable than that in the case of females, and thus, a sex difference in the reaction features may be responsible for the observed difference in the incidence of calculus formation.
Assuntos
Compostos de Bifenilo/metabolismo , Cálculos da Bexiga Urinária/metabolismo , Animais , Compostos de Bifenilo/toxicidade , Cromatografia Líquida de Alta Pressão , Microanálise por Sonda Eletrônica , Feminino , Masculino , Ratos , Ratos Endogâmicos F344 , Fatores Sexuais , Espectroscopia de Infravermelho com Transformada de Fourier , Bexiga Urinária/efeitos dos fármacos , Bexiga Urinária/metabolismo , Cálculos da Bexiga Urinária/química , Cálculos da Bexiga Urinária/patologiaRESUMO
The relationship between infection stone and Corynebacterium species was investigated in vitro and in vivo. Urease activity of urease-splitting Corynebacterium species was evaluated by 2 methods; an increase in pH of human urine after inoculation of Corynebacterium species and direct measurement of urease activity of 10(7) CFU organisms from amounts of ammonia by indophenol method. Formation of infection bladder stone was induced in male Wistar rats by implanting a zinc disc and inoculating 10(6) CFU organisms surgically into the bladder. Urine was alkalinized by the inoculation of Corynebacterium renale, C. pilosum and group D2 Corynebacterium. C. renale and C. pilosum had strong urease activity, and group D2 Corynebacterium had moderate activity. C. pseudodiphtheriticum did not produce the elevation of urinary pH and had little urease activity. Infection stones were formed in 100% of rats by inoculation of C. renale and C. pilosum and 88% of rats by group D2. Urinary pH was elevated in all inoculated rats. In conclusion, C. renale, C. pilosum and group D2 Corynebacterium may play a role in formation of infection stones.
Assuntos
Infecções por Corynebacterium , Cálculos da Bexiga Urinária/etiologia , Cálculos da Bexiga Urinária/microbiologia , Animais , Contagem de Colônia Microbiana , Corynebacterium/enzimologia , Humanos , Concentração de Íons de Hidrogênio , Masculino , Ratos , Ratos Wistar , Urease/metabolismo , Cálculos da Bexiga Urinária/metabolismo , Urina/química , Urina/microbiologiaRESUMO
Bisphosphonates bind renal calculi and inhibit calcium oxalate crystal growth in vitro. We evaluated their ability to inhibit calcium oxalate urolithiasis in a lithogenic rat model. Male Sprague-Dawley rats (four groups, eight rats each) were fed 1.0% ethylene glycol in their drinking water for 6 weeks. All rats had implantation of a 50- to 60-mg zinc pellet in their urinary bladder at the beginning of the 6-week period. The control group received no treatment. The other three groups received six weekly intraperitoneal injections of one of three bisphosphonates: pamidronate (APD), clodronate (CLO), or methylene diphosphonate (MDP). At the end of 6 weeks, the zinc pellet was retrieved and weighed; the kidneys were sectioned and stained to evaluate inflammation, tubular dilation, and crystal deposition; and blood and urine samples were analyzed for calcium and creatinine. There were no detectable biochemical differences between the control and the treatment groups. Zinc pellets removed from control animals had a significantly greater increase in weight secondary to crystal deposition than those from the treatment groups (mean 28.4% for control v 18.9%, 15.3%, and 18.6%, respectively, for animals given APD, CLO, and MDP). The control animals also had significantly higher scores for inflammation, tubular dilation, and crystal deposition than animals treated with MDP and CLO. Older and newer-generation bisphosphonates have an inhibitory effect on calcium oxalate urolithiasis that is demonstrable at relatively infrequent dosing intervals in vivo. More frequent dosing or higher doses may allow greater inhibition of stone formation.
Assuntos
Oxalato de Cálcio/antagonistas & inibidores , Ácido Clodrônico/farmacologia , Difosfonatos/farmacologia , Modelos Animais de Doenças , Cálculos da Bexiga Urinária/tratamento farmacológico , Analgésicos não Narcóticos/administração & dosagem , Analgésicos não Narcóticos/farmacologia , Analgésicos não Narcóticos/uso terapêutico , Animais , Cálcio/sangue , Cálcio/urina , Ácido Clodrônico/administração & dosagem , Ácido Clodrônico/uso terapêutico , Creatinina/sangue , Creatinina/urina , Difosfonatos/administração & dosagem , Difosfonatos/uso terapêutico , Relação Dose-Resposta a Droga , Seguimentos , Injeções Intraperitoneais , Masculino , Pamidronato , Ratos , Ratos Sprague-Dawley , Cálculos da Bexiga Urinária/induzido quimicamente , Cálculos da Bexiga Urinária/metabolismo , ZincoRESUMO
Twenty children with endemic vesical stones showed normal plasma and urinary excretion of fluoride on a mean fluoride intake of 2.5 +/- 0.8 mg/24 h. The mean fluoride content of the stones obtained from these children was 315.6 +/- 264.9 micrograms/g in the nucleus and 229.9 +/- 212.8 micrograms/g in the periphery (this was not statistically significant). Calcium-containing stones had a higher fluoride content than stones containing uric acid and ammonium urate. It was concluded that children with endemic vesical stones have normal fluoride metabolism. Trace quantities of fluoride present equally in the nucleus and peripheral parts of the stones suggest that fluoride does not cause initiation or growth of the nucleus of vesical stones and is adventitiously deposited with calcium salts in these stones.
Assuntos
Fluoretos/metabolismo , Cálculos da Bexiga Urinária/química , Cálculos da Bexiga Urinária/metabolismo , Oxalato de Cálcio/análise , Criança , Pré-Escolar , Feminino , Humanos , Magnésio/análise , Masculino , Fósforo/análise , Compostos de Amônio Quaternário/análise , Ácido Úrico/urinaRESUMO
The normal urothelium is covered by a glycosaminoglycan (GAG) layer which acts as a barrier to the adhesion of crystals. Destruction of the GAG layer increases the number of adhered crystals, and it is therefore assumed that it promotes crystal growth and stone formation. Intravesical instillation of pentosanpolysulfate, an exogenous glycosaminoglycan, after destruction of this layer reduces the adhesion of crystals to the urothelium. Intramuscular administration of carbenoxolone sodium following the experimental destruction of the GAG layer increases the rate of healing of the layer and reduces the number of adhered crystals.
Assuntos
Oxalato de Cálcio/metabolismo , Carbenoxolona/farmacologia , Ácido Glicirretínico/análogos & derivados , Poliéster Sulfúrico de Pentosana/farmacologia , Polissacarídeos/farmacologia , Bexiga Urinária/efeitos dos fármacos , Adesividade , Administração Intravesical , Animais , Carbenoxolona/administração & dosagem , Cristalização , Epitélio/efeitos dos fármacos , Epitélio/metabolismo , Epitélio/patologia , Feminino , Glicosaminoglicanos/metabolismo , Ácido Clorídrico/efeitos adversos , Injeções Intramusculares , Poliéster Sulfúrico de Pentosana/administração & dosagem , Ratos , Ratos Endogâmicos , Bexiga Urinária/metabolismo , Bexiga Urinária/patologia , Cálculos da Bexiga Urinária/metabolismo , Cálculos da Bexiga Urinária/prevenção & controleRESUMO
We studied the mechanism of gallbladder sludge formation in guinea pigs (n = 30) treated with lincomycin (80 mg/kg/day) for 7 consecutive days. At sacrifice (day 8) gallbladders of treated animals contained turbid bile, sludge and in one animal a single gallstone. The precipitates were amorphous on X-ray diffraction. Infra-red spectroscopy revealed calcium phosphate as the major component. Compared to saline-treated controls (n = 15) concentrations of total protein, total phosphate and total bilirubin in gallbladder bile were significantly increased (P less than 0.05). The increase in total phosphate was due to the inorganic component, since phospholipid phosphorus was unchanged. The relative amounts of unconjugated bilirubin and of bilirubin mono- and diconjugates in gallbladder bile were unaffected by treatment as was beta-glucuronidase activity. However, sludge was enriched in unconjugated bilirubin compared to gallbladder bile. This was most probably caused by alkaline hydrolysis of bilirubin monoconjugates. To some extent, disproportionation of bilirubin monoconjugates in bile or sludge, either in vivo or during sample preparation, might also have led to increased unconjugated pigment.
Assuntos
Colelitíase/metabolismo , Lincomicina/farmacologia , Fosfatos/metabolismo , Cálculos da Bexiga Urinária/metabolismo , Animais , Bile/análise , Bile/metabolismo , Bilirrubina/análise , Bilirrubina/metabolismo , Colelitíase/induzido quimicamente , Feminino , Cobaias , Masculino , Fosfatos/análise , Cálculos da Bexiga Urinária/análiseRESUMO
The Constant Composition (CC) kinetics method has been used for studying the mineralization of calcium oxalate monohydrate (COM) at sustained supersaturations in the presence of pre-bladder urine and macromolecules isolated from normal urine and kidney and bladder stones. The method is especially sensitive for investigating the inhibitory activities of these urinary macromolecular components (UMMC) and matrix macromolecular components (MMMC) with a coefficient of variation in growth rate of approximately 2%. Significant COM mineral inhibition was observed in a wide molecular weight region of urine components. Urine removed directly from the kidney showed appreciable inhibitory activity towards COM crystallization. Normal urinary proteins and the dissolved precipitate resulting from urine centrifugation were fractionated by gel filtration. The resulting solutions were mostly COM mineralization inhibitors. Electrodialysis was utilized to isolate the MMMC (greater than 7000 d) of renal and bladder calculi. While these solutions inhibited COM crystallization, they were also found to be calcium binders as measured by the calcium electrode.
Assuntos
Oxalato de Cálcio/antagonistas & inibidores , Glicoproteínas/urina , Cálculos Renais/metabolismo , Cálculos da Bexiga Urinária/metabolismo , Oxalato de Cálcio/urina , Precipitação Química , Cromatografia em Gel , Cristalização , Humanos , Cálculos Renais/análise , Substâncias Macromoleculares/análise , Cálculos da Bexiga Urinária/análiseRESUMO
Of 132 children with bladder stones seen in 1 year, 94% were boys and 73% were aged between 1 and 5 years. Significant malnutrition and evidence of vitamin deficiency were absent. Wheat bread was the dietary staple, while the intake of milk and dairy products, eggs and meat was very low. Analysis of 29 stones showed them to consist mainly of calcium oxalate and uric acid, with small amounts of calcium phosphate and magnesium ammonium phosphate in some cases. The composition of the surface and central parts of the stones, examined in 20 cases, was usually different. The nucleus was formed almost entirely of calcium oxalate and uric acid in four and three cases respectively, whereas the surface layers were composed of other ingredients. In another case the nucleus was composed entirely of xanthine, which indicated a diagnosis of xanthinuria.