Classification of tissue biopsies by Raman spectroscopy guided by quantitative phase imaging and its application to bladder cancer.

We present a multimodal label-free optical measurement approach for analyzing sliced tissue biopsies by a unique combination of quantitative phase imaging and localized Raman spectroscopy. First, label-free quantitative phase imaging of the entire unstained tissue slice is performed using automated scanning. Then, pixel-wise segmentation of the tissue layers is performed by a kernelled structural support vector machine based on Haralick texture features, which are extracted from the quantitative phase profile, and used to find the best locations for performing the label-free localized Raman measurements. We use this multimodal label-free measurement approach for segmenting the urothelium in benign and malignant bladder cancer tissues by quantitative phase imaging, followed by location-guided Raman spectroscopy measurements. We then use sparse multinomial logistic regression (SMLR) on the Raman spectroscopy measurements to classify the tissue types, demonstrating that the prior segmentation of the urothelium done by label-free quantitative phase imaging improves the Raman spectra classification accuracy from 85.7% to 94.7%.

Segmentation of the urothelium in optical coherence tomography images with dynamic contrast.

SIGNIFICANCE: Speckle variation induced by intracellular motion (IM) in the urothelium was observed in optical coherence tomography (OCT) images. IM can be used as a dynamic contrast to segment the urothelium by comparing two sequential OCT images. This method opens the possibility of specifically tracking the distribution of urothelial cancerous cells for identifying the microinvasion of bladder tumors. APPROACH: OCT images were acquired ex vivo with fresh porcine bladder tissue. IM was analyzed by tracking speckle variation using autocorrelation function, then quantified with constrained regularization method for inverting data (CONTIN method) to identify the decorrelation time (DT) of the speckle variations. Variance analysis was also conducted to show IM amplitude and distribution in the urothelium. The segmentation of the urothelium was demonstrated with OCT images with a visible urothelial layer and OCT images with an invisible urothelial layer. RESULTS: Significant speckle variation induced by IM was observed in the urothelium. However, the distribution of the IM is heterogeneous. The DTs are mostly concentrated between 1 and 30 ms. With the IM as a dynamic contrast, the urothelium can be accurately and exclusively segmented, even the urothelial layer is invisible in normal OCT images. CONCLUSIONS: IM can be used as a dynamic contrast to exclusively track urothelial cell distribution. This contrast may provide a new mechanism for OCT to image the invasion depth and pattern of urothelial cancerous cells for accurately substaging of bladder cancer.

A Calibrated Multiexit Neural Network for Detecting Urothelial Cancer Cells.

Deep convolutional networks have become a powerful tool for medical imaging diagnostic. In pathology, most efforts have been focused in the subfield of histology, while cytopathology (which studies diagnostic tools at the cellular level) remains underexplored. In this paper, we propose a novel deep learning model for cancer detection from urinary cytopathology screening images. We leverage recent ideas from the field of multioutput neural networks to provide a model that can efficiently train even on small-scale datasets, such as those typically found in real-world scenarios. Additionally, we argue that calibration (i.e., providing confidence levels that are aligned with the ground truth probability of an event) has been a major shortcoming of prior works, and we experiment a number of techniques to provide a well-calibrated model. We evaluate the proposed algorithm on a novel dataset, and we show that the combination of focal loss, multiple outputs, and temperature scaling provides a model that is significantly more accurate and calibrated than a baseline deep convolutional network.

High-grade Urothelial Carcinoma with Malignant Melanocytic Differentiation.

Urothelial carcinoma usually shows divergent differentiation and variant histology with squamous and glandular morphology being most common. In this report, we present a case of divergent malignant melanocytic differentiation in a high-grade urothelial carcinoma. A 98-year-old East Asian woman with an anterior bladder wall mass underwent resection, which revealed a high-grade poorly differentiated tumor. A minor component of high-grade papillary urothelial carcinoma and carcinoma in situ is also present. The majority of the tumor cells are morphologically and immunohistochemically consistent with melanoma, a minority of cells are positive for urothelial markers, and rare cells coexpress both melanocytic and urothelial markers. Cells that express melanocytic markers or urothelial markers are intimately admixed together. Taken together, a diagnosis of high-grade urothelial carcinoma with malignant melanocytic differentiation was rendered. This is the first report in the literature of malignant melanocytic differentiation in a high-grade urothelial carcinoma, a finding that may have important diagnostic and therapeutic implications.

Preoperative predictive factors of carcinoma in situ in the normal-appearing mucosa in patients who underwent an initial transurethral resection for non-muscle-invasive bladder cancer under white light cystoscopy.

BACKGROUND: Although few studies evaluated the significance of random biopsies under white light cystoscopy (WLC) in patients with non-muscle-invasive bladder cancer (NMIBC), the findings are controversial. AIM: This aim of this study was to evaluate what kind of preoperative covariates were useful as predictive factors in detecting carcinoma in situ (CIS) from normal-appearing mucosa using random bladder biopsies under WLC. METHODS AND RESULTS: A total of 229 patients with NMIBC underwent initial TUR followed by random biopsies under WLC at Red Cross Takayama Hospital between 2007 and 2016. These patients underwent TUR with complete resection of intravesical visible tumors followed by random biopsies of normal-appearing mucosa. In this study, random bladder biopsies of normal-appearing urothelial mucosa, excluding abnormal mucosa, were carried out with a cold punch in the selected intravesical sites. The covariates included age, gender, the urine cytology result, presence of an abnormal mucosa, number of tumors, size of the largest tumors, configuration of the tumor, and tumor type. Abnormal mucosa was defined as reddish or mossy areas at the time of TUR under WLC. The primary endpoint was to determine what kind of preoperative covariates were useful as predictive factors in detecting CIS from normal-appearing mucosa using random bladder biopsies under WLC. Finally, 212 patients were evaluated, and 67 patients (31.6%) were diagnosed with CIS from normal-appearing mucosa. In univariate analysis, positive urine cytology, abnormal mucosa, and the number of tumors were significantly associated with concomitant CIS. On multivariate analysis, positive urine cytology and abnormal mucosa were significantly associated with CIS. CONCLUSION: The patients who were diagnosed with positive urine cytology or abnormal mucosa by WLC are ideal candidates for TUR followed by random biopsy of normal-appearing mucosa.

Update on Renal Neoplasms: Clinicopathologic-Radiologic Correlation With Case-Based Examples.

OBJECTIVE. This article provides a brief overview of the clinicopathologic and radiologic correlation of 12 renal neoplasms, encompassing the conventional subtypes of renal cell carcinoma and a few of the newly recognized subtypes from the 2016 World Health Organization classification of renal tumors. In addition, we touch upon infrequent neoplasms that may enter the differential diagnosis of a renal mass, with corresponding radiologic and gross images and histologic findings of case-based examples. CONCLUSION. Familiarity with the radiologic and pathologic characteristics of renal cell carcinoma and other renal neoplasms is important to correctly identify and treat these masses.

Comparison of urine cytology diagnostic reports before and after the implementation of the Paris System classification system in China.

OBJECTIVE: In 2013, The Paris System for Reporting Urinary Cytology (TPS) was developed as a uniform practical urine cytology system that could be applied worldwide. Here, we investigated the effectiveness of TPS diagnostic approach compared with that of the traditional urine cytological diagnosis method used in China. METHODS: Based on the diagnostic criteria of TPS, 412 urine samples from 143 patients with histological follow-up data were retrospectively analysed, and the diagnoses were compared with the original cytological diagnoses. RESULTS: In total, 110 patients were histologically diagnosed with high-grade urothelial carcinoma (HGUC), and 33 patients were diagnosed with low-grade urothelial neoplasia. Based on the traditional urine cytological analysis method, 50 patients (34.9%) were diagnosed as negative, 48 patients (33.6%) were diagnosed as having atypical urothelial cells, and 45 patients (31.5%) were diagnosed as positive. After reclassification using TPS, urine samples from 11 cases (7.7%) were categorised as unsatisfactory, 34 cases (23.8%) were negative, 21 cases (14.7%) were categorised as having atypical urothelial cells, 12 cases (8.4%) were diagnosed as suspicious for HGUC, 59 cases (41.2%) were diagnosed with HGUC, and six cases (4.2%) were reclassified as having low-grade urothelial neoplasia. Thus, after implementing TPS criteria, the sensitivity for positive malignancy diagnoses (HGUC alone) increased from 38.2% to 50.9%, while the specificity of the diagnosis was barely changed. CONCLUSIONS: The Paris System for Reporting Urinary Cytology greatly contributes to the standardisation of urine cytology reports and significantly improves the diagnostic sensitivity for HGUC.

Sonographically Diagnosed Urothelial Thickening in Kidney Allografts: A Noninvasive and Clinically Highly Relevant Marker for the Detection of Acute Rejection.

OBJECTIVE. The objective of our study was to investigate the significance of sonographic features in assessing for acute kidney allograft rejection in the modern era. MATERIALS AND METHODS. In this retrospective study, 107 adult patients with a kidney allograft biopsy performed between 2015 and 2018 and diagnostic ultrasound performed within 2 weeks of the biopsy were included. Acute rejection was diagnosed on the basis of biopsy tissue sample results using the Banff criteria. The following ultrasound features were assessed: perfusion, cortical echogenicity, corticomedullary differentiation, urothelial thickening, change in renal length, renal artery velocity, and intraparenchymal arterial resistive index. Subjective measures of perfusion, echogenicity, corticomedullary differentiation, and urothelial thickening were assessed independently and in consensus by three abdominal radiologists; multirater kappa values were calculated for interobserver variability. The Wilcoxon rank sum test and chi-square test were used to evaluate the differences between two groups (rejection vs no rejection) and the sonographic features. Sensitivity, specificity, positive predictive value, and negative predictive value (NPV) were calculated for sonographic features that are associated with acute rejection. RESULTS. Of the sonographic features, only the presence of urothelial thickening was significantly associated with acute rejection (p < 0.001) and had substantial agreement (κ = 0.61) among readers. Urothelial thickening was highly sensitive (96%; 95% CI, 79-100%) with a high NPV (98%; 95% CI, 86-100%). CONCLUSION. Urothelial thickening on ultrasound is a highly sensitive finding for acute kidney rejection with a high NPV and thus may play a role in sonographic prebiopsy screening. Other historically associated sonographic features seem to play little, if any, role in the screening and assessment for kidney allograft rejection in the modern era.

A Rare Case of Extensive Unilateral Ureteritis Cystica.

Ureteritis cystica is rare, benign entity that associates with chronic urothelial irritation such as recurrent urinary tract infection or nephrolithiasis. It is often diagnosed incidentally on routine imaging or ureteroscopy in asymptomatic individuals. In this case report, we present the retrograde pyelogram and ureteroscopy images of a rare case of extensive unilateral ureteritis cystica in a 78-year-old female presenting for elective stone surgery.

Renal cryoablation - does deep endophytic ablation affect the renal collecting system?

Objective: To investigate to what extent the urothelium of the renal collecting system is affected when performing deep endophytic cryoablation.Methods: The study was conducted as an in vivo animal model with a total of 15 female pigs. Each animal was subjected to bilateral endophytioc renal cryoablation and randomized to a postoperative follow-up period of either one, two or four weeks. At the end of follow-up all animals had a magnetic resonance imaging (MRI) examination and bilateral nephrectomy was performed. On MRI-imaging the extent of the cryolesions, as well as signs of urinomas or fistulas, were examined. Histopathologic examinations were performed to investigate the effect on the urothelium.Results: All animals tolerated the procedure well without any postoperative complications. MRI examinations found the renal collecting system to be involved in the cryolesions at all three stages of follow-up and revealed no signs of hematomas, urinomas or fistula formations. Epithelial edema was found at all three stages of follow-up while significant parenchymal fibrosis adjacent to the urothelium was most pronounced in the four weeks of follow-up group. The urothelium was significantly affected with luminal hemorrhage as well as hemorrhage in and underneath the urothelium and urothelial dissociation from the underlying renal parenchyma. Despite these impacts on the urothelium, this was found to be intact and vital at all three stages of follow-up, in sharp contrast to the renal parenchyma that underwent fibrotic changes.Conclusions: In this, in vivo non-tumor pig model CA effectively destroyed the renal parenchyma while the impacted renal urothelium remained intact and did not undergo fibrotic changes, nor was urinomas or fistulas observed.

Assessment of 213Bi-anti-EGFR MAb treatment efficacy in malignant cancer cells with [1-13C]pyruvate and [18F]FDG.

Evaluation of response to therapy is among the key objectives of oncology. A new method to evaluate this response includes magnetic resonance spectroscopy (MRS) with hyperpolarized 13C-labelled metabolites, which holds promise to provide new insights in terms of both therapeutic efficacy and tumor cell metabolism. Human EJ28Luc urothelial carcinoma and LN18 glioma cells were treated with lethal activity concentrations of a 213Bi-anti-EGFR immunoconjugate. Treatment efficacy was controlled via analysis of DNA double-strand breaks (immunofluorescence γH2AX staining) and clonogenic survival of cells. To investigate changes in metabolism of treated cells vs controls we analyzed conversion of hyperpolarized [1-13C]pyruvate to [1-13C]lactate via MRS as well as viability of cells, lactate formation and lactate dehydrogenase activity in the cellular supernatants and [18F]FDG uptake in treated cells vs controls, respectively. Treatment of malignant cancer cells with 213Bi-anti-EGFR-MAb induced intense DNA double-strand breaks, resulting in cell death as monitored via clonogenic survival. Moreover, treatment of EJ28Luc bladder cancer cells resulted in decreased cell viability, [18F]FDG-uptake and an increased lactate export. In both EJ28Luc and LN18 carcinoma cells treatment with 213Bi-anti-EGFR-MAb triggered a significant increase in lactate/pyruvate ratios, as measured with hyperpolarized [1-13C]pyruvate. Treatment with 213Bi-anti-EGFR-MAb resulted in an effective induction of cell death in EJ28Luc and LN18 cells. Lactate/pyruvate ratios of hyperpolarized [1-13C]pyruvate proved to detect early treatment response effects, holding promise for future clinical applications in early therapy monitoring.

Is hysterectomy beneficial in radical cystectomy for female patient with urothelial carcinoma of bladder? A retrospective analysis of consecutive 112 cases from a single institution.

BACKGROUND: There is no criterion for determining whether female patients operated with cystectomy would benefit from hysterectomy. This study compares the oncological outcomes between female patients receiving uterus preserving cystectomy (UPC) and uterus excision cystectomy (UEC). METHODS: Retrospective review of 121 female patients with urothelial carcinoma of bladder undergoing UPC (n = 63) or UEC (n = 49) at a single institute between January 2006 and April 2017. Individual postoperative follow-up plans were performed for patients through outpatient visits. Overall survival (OS) and progression-free survival (PFS) estimates were analyzed using Kaplan-Meier method and multivariable Cox regression. RESULTS: The median follow-up time was 36 months (interquartile range 16-69). Among patients, 5 (4.1%) had uterus invasion. OS probability (p = 0.939) and PFS probability (p = 0.565) were similar in two groups. In multivariable Cox regression analysis, hysterectomy was not found to be a predictor of OS (hazard ratio 0.908, 95%CI 0.428-1.924, p = 0.801) and PFS (hazard ratio 1.109, 95%CI 0.439-2.805, p = 0.826) after adjusting for age, preoperative clinical stage, pathological stage, pathological nodal stage, neoadjuvant/adjuvant chemotherapy, location of the tumor, and surgical margin. No significant difference of overall survival probability was observed in the patients with organ-confined bladder cancer (p = 0.675) and in patients with no organ-confined bladder cancer (p = 0.695). CONCLUSIONS: The results showed that the rate of uterus invasion was low in patients analyzed in this cohort. It was also found that hysterectomy was not an independent predictor of OS and PFS after radical cystectomy in patients with bladder cancer.

Relapsed papillary urothelial neoplasm of low malignant potential (PUNLMP) of the young age: a case report and a review of the literature.

BACKGROUND: Papillary Urothelial Neoplasm of Low Malignant Potential (PUNLMP) are exceptionally rare in the first decade of life (mostly if multifocal) and there is a lack of standardized recommendations for the pediatric age. CASE PRESENTATION: We describe the case of a 9-year-old boy with a diagnosis of PUNLMP, who underwent to cystoscopic lesion removal and later to endoscopic lesion removal and intra-bladder Mitomycin-c (MMC) instillations for relapsed disease. Follow-up investigations at five years showed disease negativity. CONCLUSIONS: Intra-bladder MMC instillation may allow obtaining the complete remission with bladder-sparing for paediatric patients with a high-risk relapsed PUNLMP.

Urothelial Carcinoma Detected by Ultrasonography in a Renal Transplant Recipient.

Urothelial cell carcinoma in renal transplant recipients has been rarely reported. Here, we report a case of high-grade papillary urothelial carcinoma in a renal transplant recipient who presented with hematuria. Sonographic evaluations (gray-scale ultrasonography, Doppler imaging, and strain sonoelastography) helped with diagnosis in our initial approach to the patient. Computed tomography was performed to confirm solid ureteral masses and staging of the disease. The patient, who underwent nephro-ureterectomy of the graft kidney, had that kidney for 6 years, 6 months. After surgery, the patient was symptom free and without evidence of recurrence or metastasis in 3 years of follow-up. Sonographic examinations have an important role in the evaluation of urothelial carcinoma of transplanted kidneys.

Incidence and survival outcomes in patients with upper urinary tract urothelial carcinoma diagnosed with variant histology and treated with nephroureterectomy.

OBJECTIVE: To evaluate the incidence and survival outcomes of histological variants of upper tract urothelial carcinoma (UTUC) treated with radical nephroureterectomy (RNU). MATERIALS AND METHODS: We retrospectively analysed data from 1610 patients treated with RNU for clinically non-metastatic UTUC between 1990 and 2016 in several centres participating in the UTUC Collaboration. Histological variants were classified as micropapillary, squamous, sarcomatoid and other, including other rare variants (<10 cases for each). Multivariable competing risk analyses were conducted to assess the effect of variant histology on overall recurrence and cancer-specific mortality (CSM). RESULTS: Overall, 1460 patients (91%) had pure urothelial carcinoma (PUC), whereas 150 (9%) were diagnosed with a variant histology, including 89 (5.0%), 41 (2.0%), 10 (1.0%) and 10 (1.0%) cases of micropapillary, squamous, sarcomatoid and other tumours, respectively. Variant histology was associated with the presence of adverse pathological features compared with PUC, including non-organ-confined disease (59% vs 38%; P < 0.001), lymph node invasion (28% vs 24%; P = 0.02), high-grade disease (88% vs 71%; P < 0.001), tumour necrosis (28% vs 16%; P = 0.001) and positive surgical margins (15% vs 8%; P = 0.01). In competing risk analysis, micropapillary variant was the only factor associated with worse recurrence (sub-hazard ratio [SHR] 2.27, 95% confidence interval [CI] 1.25-4.79; P = 0.02) whereas sarcomatoid variant was associated with worse CSM (SHR 16.8, 95% CI 6.86-41.17; P < 0.001). CONCLUSION: We found that one out of 10 patients with UTUC treated with RNU had variant histology. Only micropapillary and sarcomatoid variants were associated with poorer oncological outcomes after adjusting for available confounding factors.