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Ureteral stones are a major clinical problem in urology that require effective and safe therapeutic options. Semirigid and flexible ureteroscopy ar two well-established procedures for treating these stones. The present review provides an outline of the advantages and disadvantages of these approaches. Semirigid ureteroscopy, which uses a rigid straight instrument, provides excellent stone visualisation and successful fragmentation. It is especially useful for proximal and bigger stones, frequently resulting in high stone-free rates and reduced procedure times. Nonetheless, its stiffness can make it difficult to navigate the delicate ureteral anatomy and increase the risk of mucosal injury. On the other hand, flexible ureteroscopy employs a more adjustable flexible scope, allowing access to complicated ureteral configurations while reducing the risk of ureteral trauma. It excels at treating distal and difficult stones but has a lower efficacy with larger stones and often requires longer procedure times. The choice between semirigid and flexible ureteroscopy is determined by patient-specific factors such as stone characteristics or anatomical considerations and the surgeon's skill. A customised approach that uses the capabilities of both treatments as needed can improve stone management outcomes while reducing potential problems. The continued advancement of technology and methodological modifications is predicted to improve the field of ureteroscopic stone management.
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Introduction: The aim of our study was to retrospectively evaluate the stone-free rate after the second intervention for every performed procedure (semirigid ureteroscopy, flexible ureteroscopy, semirigid plus flexible ureteroscopy and percutaneous nephrolithotomy). Materials and methods: A total of 149 patients, who came to the emergency room of "Saint John" Emergency Clinical Hospital, Bucharest, Romania, with acute renal pathology suggesting the presence of renal and/or ureteral lithiasis confirmed by imagining (x-ray, ultrasonography or computer tomography), were included in this retrospective study, which was conducted between September 2021 and September 2022. All selected patients had an indication of emergency surgical intervention that consisted of a double-J stent mounting. We analyzed the stone-free rate after the secondary intervention, which was one of the following procedures: semirigid ureteroscopy, flexible ureteroscopy, combined semirigid and flexible ureteroscopy (F-URS), and percutaneous nephrolithotomy (PCNL). Patients came back for the secondary intervention and were reevaluated using imaging techniques. Results:Endoscopic procedures were performed by 14 surgeons over a time period of two to six weeks after the initial stenting procedure. Encrusted stents were encountered in four cases and in five cases the patients were admitted with obstruction of previously inserted stents. From the total of 149 patients, 68 (45,6%) subjects underwent semirigid ureteroscopy, with a stone-free rate of 86% (59 cases), 32 (21,4%) F-URS, with a stone-free rate of 90,6% (30 cases), 41 (27,5%) combined flexible and semirigid ureteroscopy, with a stone-free rate of 90,24% (37 cases), and eight (5,3%) patients received PCNL, with a stone-free rate of 75% (six cases). Overall stone-free rate for all procedures was 90,06%. The mean operative time was 23 minutes. No major incidents or complications occurred during the procedures. Conclusion:The emergency pre-stenting before the definitive treatment of reno-ureteral lithiasis is a safe procedure. Flexible ureteroscopy was the most successful secondary intervention, with the highest rate of achieving stone-free status.
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OBJECTIVE: The objective of this meta-analysis is to evaluate the impact of genetic polymorphisms on platinum-based chemotherapy (PBC)-induced ototoxicity. DATA SOURCES: Systematic searches of PubMed, Embase, Cochrane, and Web of Science were conducted from the inception of the databases to May 31, 2022. Abstracts and presentations from conferences were also reviewed. REVIEW METHODS: Four investigators independently extracted data in adherence to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines. Differences in the prevalence of PBC-induced ototoxicity between reference and variant (i) genotypes and (ii) alleles were analyzed. The overall effect size was presented using the random-effects model as an odds ratio (OR) with a 95% confidence interval (CI). RESULTS: From 32 included articles, 59 single nucleotide polymorphisms on 28 genes were identified, with 4406 total unique participants. For allele frequency analysis, the A allele in ACYP2 rs1872328 was positively associated with ototoxicity (OR: 2.61; 95% CI: 1.06-6.43; n = 2518). Upon limiting to cisplatin use only, the T allele of COMT rs4646316 and COMT rs9332377 revealed significant results. For genotype frequency analysis, the CT/TT genotype in ERCC2 rs1799793 demonstrated an otoprotective effect (OR: 0.50; 95% CI: 0.27-0.94; n = 176). Excluding studies using carboplatin or concomitant radiotherapy revealed significant effects with COMT rs4646316, GSTP1 rs1965, and XPC rs2228001. Major sources of variations between studies include differences in patient demographics, ototoxicity grading systems, and treatment protocols. CONCLUSION: Our meta-analysis presents polymorphisms that exert ototoxic or otoprotective effects in patients undergoing PBC. Importantly, several of these alleles are observed at high frequencies globally, highlighting the potential for polygenic screening and cumulative risk evaluation for personalized care.
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Antineoplásicos , Ototoxicidade , Humanos , Antineoplásicos/uso terapêutico , Ototoxicidade/tratamento farmacológico , Platina , Cisplatino , Polimorfismo de Nucleotídeo Único , Proteína Grupo D do Xeroderma Pigmentoso/genética , Hidrolases Anidrido Ácido/genéticaRESUMO
Objective: Ureteroscopy (URS) is a commonly used procedure for the management of ureteral stones. While elective URS has been extensively studied, the literature on emergency URS remains limited. The aim of the present study is to evaluate the efficacy and safeness of URS performed in emergency settings and to determine the ideal candidates for this type of intervention. Methods:Patients who underwent URS for ureteral stones in a single healthcare unit, "Saint John" Emergency Clinical Hospital, Bucharest, Romania, were included in a retrospective investigation between April 2022 and March 2023. The study group was divided into two subgroups: group A (138 patients who underwent semirigid URS in an emergency setting) which was subdivided into group A1 (95 patients with stone on the distal ureter) and group A2 (43 patients with stone on the proximal ureter), and group B (151 patients who underwent a double J stent insertion). The success rate defined as stone fragmentation and resolution of renal obstruction, along with intraoperative and postoperative complications were assessed. A URS procedure was considered unsuccessful if either the instrument could not be passed to access the stone or it was deemed unsafe to perform the URS. In such cases, patients were managed by inserting a ureteric stent and scheduled for a subsequent procedure. Results:It could be observed that most complications occurred in emergency ureteroscopy on distal ureter (95 cases) and the most severe ones on proximal ureter (two cases - Clavien 4). Double J stenting provided a reduced number of complications (51 cases). It should be mentioned that patients with emergency semirigid ureteroscopy had more complications than those with double J stent for every group of BMI, while most of the complications were observed in the groups with the highest BMI. The success of the URS procedure was determined based on complete stone fragmentation and extraction, and it was of 91.3% for cases with emergency ureteroscopy. Conclusion:Patients who underwent URS for ureteral stones at a single facility, "Saint John" Emergency Clinical Hospital, Bucharest, Romania, were included in a retrospective investigation. The success rates and complication rates of emergency URS were comparable to those of elective URS, providing valuable insights for clinical decision-making.
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Introduction: The COVID-19 pandemic definitely changed the management of patients with benign prostatic hyperplasia (BPH). This study followed the modalities of treatments in patients with BPH associated with SARS-CoV-2 attending the Urology Clinic of "Sf. Ioan" Emergency Clinical Hospital, Bucharest, Romania. Material and methods:The present study included 81 patients (mean age 63.2 years, age range 55-87 years) with SARS-CoV-2 and BPH who were admitted to our Urology Department between January 2021 and January 2022. The diagnosis of SARS-CoV-2 was based on the PCR test and that of BPH by using the diagnostic triad consisting of digital rectal examination, PSA, free PSA and ultrasound examination. It should be noted that some of the hospitalized patients were following treatment with alpha blockers and/or 5-alpha-reductase inhibitors at the time of admission. Results:Out of the 81 hospitalized cases, 13 required emergency endoscopic intervention under spinal anaesthesia (TURP or TURisP) for haemostasis because those patients presented with persistent haematuria which did not respond to conservative treatment. A number of 17 cases showed acute urinary retention during hospitalization and a urethrovesical catheter was fitted and will be re-evaluated urologically after the COVID episode. Of the remaining 51 subjects with BPH, 17 already had chronic urinary retention on admission, with urethrovesical probe present, 13 cases began during hospitalization with alpha-blocker treatment associated with 5-alpha-reductase inhibitors; meanwhile, there were no urological interventions to modify the treatment regimen in the remaining 21 patients, who were strictly managed on the side of COVID-19 infection. Conclusion:There was no clear influence of the evolution of patients with BPH due to SARS-CoV-2 pathology, and the general management trend was to delay chronic cases until the time of viral infection remission.
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Tumor-associated macrophages (TAMs) usually express an M2 phenotype, which enables them to perform immunosuppressive and tumor-promoting functions. Reprogramming these TAMs toward an M1 phenotype could thwart their pro-cancer activities and unleash anti-tumor immunity, but efforts to accomplish this are nonspecific and elicit systemic inflammation. Here we describe a targeted nanocarrier that can deliver in vitro-transcribed mRNA encoding M1-polarizing transcription factors to reprogram TAMs without causing systemic toxicity. We demonstrate in models of ovarian cancer, melanoma, and glioblastoma that infusions of nanoparticles formulated with mRNAs encoding interferon regulatory factor 5 in combination with its activating kinase IKKß reverse the immunosuppressive, tumor-supporting state of TAMs and reprogram them to a phenotype that induces anti-tumor immunity and promotes tumor regression. We further establish that these nanoreagents are safe for repeated dosing. Implemented in the clinic, this immunotherapy could enable physicians to obviate suppressive tumors while avoiding systemic treatments that disrupt immune homeostasis.
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Macrófagos/imunologia , Nanopartículas , Neoplasias/imunologia , RNA Mensageiro/administração & dosagem , Animais , Linhagem Celular Tumoral , Reprogramação Celular , Feminino , Glioblastoma/imunologia , Glioblastoma/patologia , Glioblastoma/terapia , Humanos , Quinase I-kappa B/genética , Quinase I-kappa B/metabolismo , Terapia de Imunossupressão , Fatores Reguladores de Interferon/genética , Fatores Reguladores de Interferon/metabolismo , Ativação de Macrófagos , Melanoma Experimental/imunologia , Melanoma Experimental/patologia , Melanoma Experimental/terapia , Camundongos , Metástase Neoplásica , Neoplasias/genética , Neoplasias/patologia , Neoplasias Ovarianas/imunologia , Neoplasias Ovarianas/patologia , Neoplasias Ovarianas/terapia , RNA Mensageiro/genética , Linfócitos T/imunologia , Fatores de Transcrição/genética , TransfecçãoRESUMO
Glucosteroids (GS) are widely used drugs for various inflammatory pathologies (Nephrotic syndrome, Proliferative glomerulonephritis, Extramembrane glomerulonephritis, Nephropathy of the Nodous Poliarterita (PAN), Nephropathy from purple Henoch-Schonlein, lupus nephropathy (LN), Acute adrenal insufficiency Waterhouse-Friederichsen, Chronic adrenal insufficiency Addison, Systemic Lupus Erythematosus (SLE), Polymyositis and dermatomyositis, Chronic granulomatosis, Crohn's disease, Hemorrhagic rectocolitis, Hemolytic anemias, Acute leukemias and chronic lymphocytic leukemia, Hodgkin's lymphoma). Although they are prescribed for their anti-inflammatory and immunosuppressive properties, they also have many side effects, hyperglycemia being one of the most common and representative, which is why these drugs need careful monitoring when administered over the long term. This paper presents the case of a 39 year old patient diagnosed with systemic lupus erythematosus (SLE) with class IV lupus nephropathy (LN) who developed numerous complications due to the pathogenic side effects: diabetes, amenorrhea, recurrent infections, and depression.
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BACKGROUND: A retrospective study was aimed to establish the eventual impact of narrow band imaging (NBI) cystos copy over the short term residual tumorsâ rate determined during the first follow-up endoscopic control in newly determined non-muscle invasive bladder cancer (NMIBC) cases. METHODS: 68 patients were found with NMIBC by means of both white light cystoscopy (WLC) and NBI. A follow-up using both investigation modalities was performed at 3 months from the initial procedure in 67 cases (one was lost from follow-up). RESULTS concerning the short termer currences were retrospectively compared to those obtain ending a similar series of 67 patients previously diagnosed and treated by means of classical WLC and resection only. RESULTS: The short term residual tumorsâ rate established during the first follow-up cystoscopy was lower in the study group initially benefiting from the NBI mode by comparison to the WLC control series (4.5% versus 11.9%). When drawing a parallel with the tumor map location outlined during the initial procedure, the most important differences were emphasized concerning other site recurrences (3%versus 8.95%). CONCLUSIONS: NBI cystoscopy and resection displayed a substantially favorable influence over the short termoncologic outcome in newly diagnosed NMIBC cases when compared to the standard protocol. ABBREVIATIONS: NBI â" narrow band imaging; NMIBC â"non-muscle invasive bladder cancer; WLC â" white light cystoscopy; TURBT â" transurethral resection of bladdertumors; CIS â" carcinoma in situ.
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Carcinoma in Situ/cirurgia , Carcinoma de Células de Transição/cirurgia , Cistoscopia , Imagem de Banda Estreita , Recidiva Local de Neoplasia/cirurgia , Neoplasias da Bexiga Urinária/cirurgia , Carcinoma in Situ/diagnóstico , Carcinoma de Células de Transição/diagnóstico , Cistoscopia/métodos , Seguimentos , Humanos , Imagem de Banda Estreita/métodos , Recidiva Local de Neoplasia/diagnóstico , Estudos Retrospectivos , Resultado do Tratamento , Neoplasias da Bexiga Urinária/diagnósticoRESUMO
OBJECTIVES: A prospective study was performed aiming to evaluate the surgical efficacy, perioperative safety profile, diagnostic accuracy and medium term results of a multi-modal approach consisting in narrow band imaging (NBI) cystoscopy and bipolar plasma vaporization (BPV) when compared to the standard protocol represented by white light cystoscopy (WLC) and transurethral resection of bladder tumors (TURBT). MATERIALS & METHODS: A total of 260 patients with apparently at least one bladder tumor over 3 cm were included in the trial. In the first group, 130 patients underwent conventional and NBI cystoscopy followed by BPV, while in a similar number of cases of the second arm, classical WLC and TURBT were applied. In all non-muscle invasive bladder tumors' (NMIBT) pathologically confirmed cases, standard monopolar Re-TUR was performed at 4-6 weeks after the initial intervention, followed by one year' BCG immunotherapy. The follow-up protocol included abdominal ultrasound, urinary cytology and WLC, performed every 3 months for a period of 2 years. RESULTS: The obturator nerve stimulation, bladder wall perforation, mean hemoglobin level drop, postoperative bleeding, catheterization period and hospital stay were significantly reduced for the plasma vaporization technique by comparison to conventional resection. Concerning tumoral detection, the present data confirmed the NBI superiority when compared to standard WLC regardless of tumor stage (95.3% vs. 65.1% for CIS, 93.3% vs. 82.2% for pTa, 97.4% vs. 94% for pT1, 95% vs. 84.2% overall). During standard Re-TUR the overall (6.3% versus 17.4%) and primary site (3.6% versus 12.8%) residual tumors' rates were significantly lower for the NBI-BPV group. The 1 (7.2% versus 18.3%) and 2 (11.5% versus 25.8%) years' recurrence rates were substantially lower for the combined approach. CONCLUSIONS: NBI cystoscopy significantly improved diagnostic accuracy, while bipolar technology showed a higher surgical efficiency, lower morbidity and faster postoperative recovery. The combined technique offered a reduced rate of residual tumors at Re-TUR, both globally as well as for orthotopic tumors. Substantially lower recurrence rates were found at 1 and 2 years among the NBI-BPV cases.
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Cistoscopia/métodos , Endoscopia Gastrointestinal/métodos , Imagem de Banda Estreita/métodos , Neoplasias da Bexiga Urinária/diagnóstico , Neoplasias da Bexiga Urinária/cirurgia , Terapia Combinada , Humanos , Estudos Prospectivos , VolatilizaçãoRESUMO
OBJECTIVES: A single centre, retrospective trial was performed trying to assess the impact of NBI cystoscopy in cases of non-muscle invasive bladder tumors (NMIBT) by comparison to the standard approach. Our goal was to determine the superiority of the new method in terms of detection rates and subsequent postoperative treatment changes. MATERIALS AND METHODS: A total of 320 NMIBT suspected consecutive cases were enrolled in the study. The inclusion criteria were represented by hematuria, positive urinary cytology and/or ultrasound suspicion of bladder tumors. All patients underwent WLC and NBI cystoscopy. Standard transurethral resection of bladder tumors (TURBT) was performed for all lesions visible in WL and NBI guided resection for solely NBI observed tumors. RESULTS: The overall NMIBT and CIS patients' detection rates were significantly improved for the NBI evaluation ((94.9% versus 88.1% and 95.7% versus 65.2%). Also, on a lesions' related basis, NBI cystoscopy emphasized a significantly superior diagnostic accuracy concerning the CIS, pTa and overall NMIBT formations ((95.2% versus 60.3%, 92.8% versus 83.9% and 94.1% versus 82%). Additional tumors were diagnosed by NBI in a significantly higher proportion of CIS, pTa, pT1 and NMIBT patients (56.6% versus 8.7%, 28% versus 10.3%, 30.3% versus 10.6% and 31.6% versus 9.4%). As a result of these supplementary findings, the postoperative treatment was significantly improved in a substantial proportion of cases (15.4% versus 5.1%). CONCLUSIONS: NBI cystoscopy represents a valuable diagnostic alternative in NMIBT patients, with significant improvement of tumor visual accuracy as well as detection rates. This approach provided a substantial amelioration to the risk category stratification and subsequent bladder cancer therapeutic management.
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Cistoscopia/métodos , Imagem de Banda Estreita/métodos , Neoplasias da Bexiga Urinária/diagnóstico , Urologia/métodos , Feminino , Humanos , Masculino , Estudos Retrospectivos , Urologia/tendênciasRESUMO
INTRODUCTION: A long term, retrospective study was performed aiming to outline a critical comparison concerning the efficacy, safety and durability of the bipolar plasma vaporization (BPV), standard monopolar transurethral resection (TUR) and "cold-knife" "star" transurethral incision (TUI) in secondary bladder neck sclerosis (BNS) cases. MATERIALS & METHODS: Of the 126 patients included in the trial based on maximum flow rate (Qmax) below 10 mL/s and International Prostate Symptom Score (IPSS) over 19, classical resection was performed in 46 cases, "cold-knife" TUI in 37 cases and bipolar vaporization in 43 patients. The evaluation protocol comprised IPSS, QoL (quality of life) score, Qmax and PVR (post-voiding residual urinary volume) assessment performed at 1, 3, 6, 12, 18 and 24 months after the initial intervention. RESULTS: Significant intraoperative complications (capsular perforation - 8.7%; bleeding - 4.3%) occurred secondary to monopolar resection. "Star" TUI was the fastest technique, followed by plasma-button vaporization (7.2 and 11.4 versus 16.5 minutes). BPV and TUI patients benefitted from the shortest catheterization periods (0.75 and 1 versus 2.0 days) and hospital stays (1.0 and 1.25 versus 2.0 days). Immediate postoperative adverse events consisted of hematuria (6.5% of the TUR cases) and acute urinary retention (8.1% of the TUI group). Significantly higher long term BNS recurrence rates requiring re-treatment were established in the TUI (18.7%) and TUR (12.8%) series by comparison to BPV (5.4%). Among patients that completed the follow-up protocol, equivalent IPSS, QoL, Qmax and PVR features were determined in the 3 study arms. CONCLUSIONS: The plasma vaporization approach was confirmed as a successful match to conventional TUR and "cold-knife" TUI in terms of surgical safety profile, postoperative recovery, therapeutic durability and urodynamic and symptom score parameters.
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Cistotomia/métodos , Esclerose/cirurgia , Ressecção Transuretral da Próstata/efeitos adversos , Ureteroscopia/métodos , Doenças da Bexiga Urinária/cirurgia , Medicina Baseada em Evidências , Humanos , Qualidade de Vida , Estudos Retrospectivos , Esclerose/etiologia , Doenças da Bexiga Urinária/etiologia , VolatilizaçãoRESUMO
INTRODUCTION: Secondary bladder neck sclerosis (BNS) represents a common late complication of prostate surgery, however so far insufficiently assessed in the available literature. More over, the previously attempted and analyzed therapeutic modalities failed to achieve acknowledgement as standard treatment for this particular pathology. METHODS: The bipolar plasma vaporization (BPV) was introduced as a viable mean of removing the obstructing scar formation in a gradual fashion. Several "tips and tricks" were described as particularly useful in optimizing the plasma corona vaporization effect. The proper BPV technique is simple and safe while closely relying on certain surgical steps, the simultaneous vaporization and coagulation processes and a superior endoscopic vision. Recent technological advances created the premises for further improvements. RESULTS: The plasma-button vaporization is characterized by a satisfactory surgical speed, remote intraoperative bleeding risks, high-quality endoscopic visibility as well as the achievement of a complete sclerotic tissue removal. Within a short learning curve, a superior final aspect of the prostatic fossa and bladder neck is obtained at the end of surgery. The continuous plasma vaporization mode provides additional technical advantages. The previously described drawbacks of transurethral incision or resection seem to have been overcome by the practical features of the plasma vaporization process. CONCLUSIONS: The BPV technique outlines a promising modality of efficiently ablating the obstructing fibrous tissue in secondary BNS patients. The simplicity and safety of the bipolar vaporization approach, together with the thorough obstructing scar removal in a radical fashion, create the premises for a favorable long term BPV clinical outcome.
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Terapia a Laser/métodos , Gases em Plasma/uso terapêutico , Bexiga Urinária/cirurgia , Perda Sanguínea Cirúrgica/prevenção & controle , Cicatriz/patologia , Diafragma/cirurgia , Humanos , Masculino , Próstata/cirurgia , EscleroseRESUMO
Abnormalities in iron metabolism are frequent in the neoplastic disease. The relationship between hepcidin and iron homeostasis in cancerous pathology is incompletely known, although it has been studied during the last years. This paper aims to analyze the role of hepcidin in the neoplastic processes, its correlation with carcinogenesis and anemia, and with the disease activity. It must be mentioned that most of the aspects presented need to be verified in practice. Insufficient data are known for showing hepcidin involvement in carcinogenesis, metastasis or in appreciating the response to anemia treatment in neoplasia.
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Hepcidinas/metabolismo , Neoplasias/metabolismo , Biomarcadores Tumorais/metabolismo , Hepcidinas/biossíntese , Hepcidinas/genética , Humanos , Ferro/metabolismo , Neoplasias/patologiaRESUMO
INTRODUCTION: The standard non-muscle invasive bladder cancer (NMIBC) endoscopic diagnosis suffers from the frequently unsatisfactory white light evaluation accuracy leading to residual lesions being left behind. Monopolar transurethral resection of bladder tumors (TURBT) is marked by a substantial morbidity rate requiring further amelioration. METHODS: Small size tumors (under 1 cm) are feasible for "en bloc" resection. Bipolar TURBT was defined as being able to challenge the gold-standard status of monopolar resection due to the reduced complication rates. Plasma-button vaporization was introduced as a promising large bladder tumors' ablation modality. Narrow band imaging (NBI) cystoscopy became an increasingly popular method of enhancing tumor detection. RESULTS: The "en bloc" resection of small size or thin pedicle tumors provides the conditions for avoiding tumoral tissue scattering. Bipolar resection is characterized by decreased perioperative bleeding risks and faster patient recovery. Plasma-button vaporization gained confirmation as an innovative approach, able to dispose large tumor bulks under complete control while minimizing the associated morbidity. NBI cystoscopy is a useful tool in identifying CIS lesions, small papillary tumors or extended margins of large tumor formations. As a cost-free technique, it may be extensively used both during the NMIBC initial diagnostic as well as during follow-up evaluation protocol. CONCLUSIONS: Having in mind the various modalities of ameliorating the bladder cancer diagnostic and treatment, NMIBC management should be tailored in accordance with the particularities of each case.
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Cistoscopia/métodos , Eletrocirurgia/métodos , Neoplasias da Bexiga Urinária/cirurgia , HumanosRESUMO
Nanostructures such as nanospheres or nanowires may contain surfaces and interfaces of pronounced curvature. To investigate the impact of severe curvature on the kinetic of reactions, thin-film Al/Cu/Al and Cu/Al/Cu triple layers are deposited on tungsten tips of 25nm curvature radius. The thermal reaction of the layer structure is studied by atom probe tomography. Experiments demonstrate that the reaction rate depends significantly on the deposition sequence of metals. Interpretation of the observed reaction kinetics leads to the conclusion that under the influence of interfacial tension probably the two limiting cases of atomic transport, Darken and Nernst-Planck kinetics, are realized in dependence on the stacking sequence.