Whole-mount Histology Research Articles

Quantification of fibrosis in infarcted swine hearts by ex vivo late gadolinium-enhancement and diffusion-weighted MRI methods

Many have speculated that MRI signal characteristics can be used to identify regions of heterogeneous infarct associated with an arrhythmogenic substrate; however, direct evidence of this relationship is limited. The aim of this study was to demonstrate the remodelling characteristics of fibrosis by means of histology and high-resolution MR imaging. For this purpose, we performed whole-mount histology in heart samples (n = 9) collected from five swine at six weeks post-infarction and compared the extent of fibrosis in the infarcted areas delineated in these histological images with that obtained ex vivo by MRI using late gadolinium-enhancement (LGE) and diffusion-weighted imaging (DWI) methods. All MR images were obtained at a submillimetre resolution (i.e., voxel size of 0.6×0.6×1.2 mm3). Specifically, in the histology images, we differentiated moderate fibrosis (consisting of a mixture of viable and non-viable myocytes, known as border zone, BZ) from severe fibrosis (i.e., the dense scar). Correspondingly, tissue heterogeneities in the MR images were categorized by a Gaussian mixture model into healthy, BZ and scar. Our results showed that (a) both MRI methods were capable of qualitatively distinguishing sharp edges between dense scar and healthy tissue from regions of heterogeneous BZ; (b) the BZ and dense scar areas had intermediate-to-high increased values of signal intensity in the LGE images and of apparent diffusion coefficient in the DWI, respectively. In addition, as demonstrated by the Picrosirius Red and immunohistochemistry stains, the viable bundles in the BZ were clearly separated by thin collagen strands and had reduced expression of Cx43, whereas the core scar was composed of dense fibrosis. A quantitative analysis demonstrated that the comparison between BZ/scar extent in LGE and DWI to the corresponding areas identified in histology yielded very good correlations (i.e., for the scar identified by LGE, R2 was 0.96 compared to R2 = 0.93 for the scar identified in ADC maps, whereas the BZ had R2 = 0.95 for the correlation between LGE and histology compared to R2 = 0.91 obtained for ADC). This novel study represents an intermediate step in translating such research to the in vivo stages, as well as in establishing the best and most accurate MR method to help identify arrhythmia substrate in patients with structural heart disease.

Abstract 5081: Development and characterization of a new antiangiogenic peptide targeting tumor vessels.

Abstract Vascular targeting is a promising approach for the treatment of angiogenesis-dependent diseases particularly for tumor treatment. Targeted drug-delivery to tumor vasculature preferentially localizes the drug at the tumor site, thus increasing efficacy and decreasing systemic side effects. In tumor neovasculature activated endothelial cells have been shown to express molecules characteristic of angiogenic vessels and virtually not expressed in normal vessels. Notably, tumor-associated endothelial cells express an isoform of aminopeptidase N (CD13) that binds to the cyclic peptide Cys-Asn-Gly-Arg-Cys (named NGR). Such “vascular addresses” allow the selective targeting of systemically administrated therapies to tumors, including chemotherapeutic agents and cytokines. In particular, the NGR-TNF conjugate has been shown to induce potent antitumor effects and is currently tested in phase II-III clinical trials. In this context, we developed and characterized a new tumor targeting peptide with a pathway of specificity comparable to that of NGR, but with a higher stability and affinity for the tumor vessels. In particular, we synthesized peptides containing the Cys-Arg-Gly-Asn-Cys cyclic motif made of D-aminoacids (i.e. crGnc, named rGn). The rGn peptide can be considered a partial retro-inverso of the NGR peptide, since a real retro-inverso petpide requires cyclization through the backbone, whereas in this case cyclization was obtained via the formation of the disulphide bond between the N and the C- terminal cysteins. The main advantage of retro-inverso peptides is their high metabolic stability, because the peptide bonds generated by D-aminoacids are stable to enzymatic cleavage. To characterize the biological activity and functions of the new peptide, we studied the rGn/CD13 interactions in vitro and in vivo on both tumour blood vessels and normal tissues, using quantum dots (Qd), whole-mount histology, and fluorescence/confocal microscopy. rGn-Qd showed the same recognition pattern of NGR-Qd on primary cell cultures of endothelial origin. Consistently, in vivo administered rGn-Qd and NGR-Qd bind to tumor blood vessels of several murine tumours. Of note, the binding of either rGn-Qd or NGR-Qd to CD13 is efficiently competed by an excess of either NGR or rGn peptide, respectively, both in in vitro and in vivo models. In particular, the rGn peptide showed a significantly higher affinity than NGR for its target cells in vitro. Moreover, silencing CD13 with specific shRNA prevented not only anti-CD13 MoAb binding, but also rGn-Qd binding. Finally, stressed experimental conditions that allowed deamidation of NGR to isoDGR, do not affect rGn structural integrity. In conclusion, the rGn peptide binds to CD13 expressed by angiogenic vessels with higher affinity and stability than NGR. Coupling rGn to cytotoxic/cytostatic molecules could lead to the generation of new potent antitumor compounds. Citation Format: Paola Di Matteo, Claudia Asperti, Simona Porcellini, Andrea Spitaleri, Claudio Bordignon, Giovanna Musco, Gian-Paolo Rizzardi, Catia Traversari. Development and characterization of a new antiangiogenic peptide targeting tumor vessels. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5081. doi:10.1158/1538-7445.AM2013-5081

Registration of prostate histology images to ex vivo MR images via strand‐shaped fiducials

To present and evaluate a method for registration of whole-mount prostate digital histology images to ex vivo magnetic resonance (MR) images. Nine radical prostatectomy specimens were marked with 10 strand-shaped fiducial markers per specimen, imaged with T1- and T2-weighted 3T MRI protocols, sliced at 4.4-mm intervals, processed for whole-mount histology, and the resulting histological sections (3-5 per specimen, 34 in total) were digitized. The correspondence between fiducial markers on histology and MR images yielded an initial registration, which was refined by a local optimization technique, yielding the least-squares best-fit affine transformation between corresponding fiducial points on histology and MR images. Accuracy was quantified as the postregistration 3D distance between landmarks (3-7 per section, 184 in total) on histology and MR images, and compared to a previous state-of-the-art registration method. The proposed method and previous method had mean (SD) target registration errors of 0.71 (0.38) mm and 1.21 (0.74) mm, respectively, requiring 3 and 11 hours of processing time, respectively. The proposed method registers digital histology to prostate MR images, yielding 70% reduced processing time and mean accuracy sufficient to achieve 85% overlap on histology and ex vivo MR images for a 0.2 cc spherical tumor.

Introducing Parametric Fusion PET/MRI of Primary Prostate Cancer

We assessed the performance of parametric fusion PET/MRI based on (11)C-choline PET/CT and apparent diffusion coefficient (ADC) maps derived from diffusion-weighted MRI for the identification of primary prostate cancer. (11)C-choline PET/CT and MRI were performed in 17 patients with untreated primary prostate cancer, followed by prostatectomy. Registration of in vivo imaging with histology was achieved using a mutual-information objective function and by performing ex vivo MRI of the prostatectomy specimen (obtained at 3 T) and whole-mount sectioning with block-face photography as intermediate steps. Data analysis included volumetrically registered whole-mount histology with Gleason scoring, (11)C-choline, and ADC data (obtained at 1.5 T). Volumes of interest were defined on the basis of histologically proven tumor tissue to calculate tumor-to-benign prostate background ratios (TBRs) for (11)C-choline, ADC, and a derived fusion PET/MRI parameter calculating the quotient of (11)C-choline over ADC (P(CHOL/ADC)). Fifty-one tumor nodules were identified at pathology. The TBRs for (11)C-choline (P < 0.05) and P(CHOL/ADC) (P < 0.005) were significantly higher in prostate cancers with a Gleason score of ≥3 + 4 than with a Gleason score of ≤3 + 3 disease and controls. For Gleason ≥ 3 + 4, the ADC TBRs were significantly lower than controls and Gleason ≤ 3 + 3 disease (P < 0.05). The absolute value of TBRs obtained from Gleason ≥ 3 + 4 cancers increased from ADC to (11)C-choline PET/CT and from (11)C-choline PET/CT to P(CHOL/ADC), with each step being statistically significant. Our data indicate that parametric PET/MRI using P(CHOL/ADC) improves lesion-to-background contrast (TBRs) of Gleason ≥ 3 + 4 disease, compared with (11)C-choline PET/CT or diffusion-weighted MRI, and thus hold promise that parametric imaging performed on hybrid PET/MRI may further improve identification and localization of significant primary prostate cancer.

Construction of 3D MR image-based computer models of pathologic hearts, augmented with histology and optical fluorescence imaging to characterize action potential propagation

Cardiac computer models can help us understand and predict the propagation of excitation waves (i.e., action potential, AP) in healthy and pathologic hearts. Our broad aim is to develop accurate 3D MR image-based computer models of electrophysiology in large hearts (translatable to clinical applications) and to validate them experimentally. The specific goals of this paper were to match models with maps of the propagation of optical AP on the epicardial surface using large porcine hearts with scars, estimating several parameters relevant to macroscopic reaction-diffusion electrophysiological models. We used voltage-sensitive dyes to image AP in large porcine hearts with scars (three specimens had chronic myocardial infarct, and three had radiofrequency RF acute scars). We first analyzed the main AP waves' characteristics: duration (APD) and propagation under controlled pacing locations and frequencies as recorded from 2D optical images. We further built 3D MR image-based computer models that have information derived from the optical measures, as well as morphologic MRI data (i.e., myocardial anatomy, fiber directions and scar definition). The scar morphology from MR images was validated against corresponding whole-mount histology. We also compared the measured 3D isochronal maps of depolarization to simulated isochrones (the latter replicating precisely the experimental conditions), performing model customization and 3D volumetric adjustments of the local conductivity. Our results demonstrated that mean APD in the border zone (BZ) of the infarct scars was reduced by ~13% (compared to ~318 ms measured in normal zone, NZ), but APD did not change significantly in the thin BZ of the ablation scars. A generic value for velocity ratio (1:2.7) in healthy myocardial tissue was derived from measured values of transverse and longitudinal conduction velocities relative to fibers direction (22 cm/s and 60 cm/s, respectively). The model customization and 3D volumetric adjustment reduced the differences between measurements and simulations; for example, from one pacing location, the adjustment reduced the absolute error in local depolarization times by a factor of 5 (i.e., from 58 ms to 11 ms) in the infarcted heart, and by a factor of 6 (i.e., from 60 ms to 9 ms) in the heart with the RF scar. Moreover, the sensitivity of adjusted conductivity maps to different pacing locations was tested, and the errors in activation times were found to be of approximately 10-12 ms independent of pacing location used to adjust model parameters, suggesting that any location can be used for model predictions.

Multimodal wavelet embedding representation for data combination (MaWERiC): integrating magnetic resonance imaging and spectroscopy for prostate cancer detection

Recently, both Magnetic Resonance (MR) Imaging (MRI) and Spectroscopy (MRS) have emerged as promising tools for detection of prostate cancer (CaP). However, due to the inherent dimensionality differences in MR imaging and spectral information, quantitative integration of T(2) weighted MRI (T(2)w MRI) and MRS for improved CaP detection has been a major challenge. In this paper, we present a novel computerized decision support system called multimodal wavelet embedding representation for data combination (MaWERiC) that employs, (i) wavelet theory to extract 171 Haar wavelet features from MRS and 54 Gabor features from T(2)w MRI, (ii) dimensionality reduction to individually project wavelet features from MRS and T(2)w MRI into a common reduced Eigen vector space, and (iii), a random forest classifier for automated prostate cancer detection on a per voxel basis from combined 1.5 T in vivo MRI and MRS. A total of 36 1.5 T endorectal in vivo T(2)w MRI and MRS patient studies were evaluated per voxel by MaWERiC using a three-fold cross validation approach over 25 iterations. Ground truth for evaluation of results was obtained by an expert radiologist annotations of prostate cancer on a per voxel basis who compared each MRI section with corresponding ex vivo wholemount histology sections with the disease extent mapped out on histology. Results suggest that MaWERiC based MRS T(2)w meta-classifier (mean AUC, μ = 0.89 ± 0.02) significantly outperformed (i) a T(2)w MRI (using wavelet texture features) classifier (μ = 0.55 ± 0.02), (ii) a MRS (using metabolite ratios) classifier (μ = 0.77 ± 0.03), (iii) a decision fusion classifier obtained by combining individual T(2)w MRI and MRS classifier outputs (μ = 0.85 ± 0.03), and (iv) a data combination method involving a combination of metabolic MRS and MR signal intensity features (μ = 0.66 ± 0.02).

High-resolution MRI of excised human prostate specimens acquired with 9.4T in detection and identification of cancers: Validation of a technique

To evaluate feasibility of high-resolution, high-field ex vivo prostate magnetic resonance imaging (MRI) as an aid to guide pathologists' examination and develop in vivo MRI methods. Unfixed excised prostatectomy specimens (n = 9) were obtained and imaged immediately after radical prostatectomy under an Institutional Review Board-approved protocol. High-resolution T2-weighted (T2W) MRI of specimens were acquired with a Bruker 9.4 T scanner to correlate with whole-mount histology. Additionally, T2 and apparent diffusion coefficient (ADC) maps were generated. By visual inspection of the nine prostate specimens imaged, high-resolution T2W MRI showed improved anatomical detail compared to published low-resolution images acquired at 4 T as published by other investigators. Benign prostatic hyperplasia, adenocarcinomas, curvilinear duct architecture distortion due to adenocarcinomas, and normal radial duct distribution were readily identified. T2 was ≈10 msec longer (P < 0.03) and the ADC was ≈1.4 times larger (P < 0.002) in the normal peripheral zone compared to the peripheral zone with prostate cancer. Differences in T2 and ADC between benign and malignant tissue are consistent with in vivo data. High-resolution, high-field MRI has the potential to improve the detection and identification of prostate structures. The protocols and techniques developed in this study could augment routine pathological analysis of surgical specimens and guide treatment of prostate cancer patients.

Elastic registration of multimodal prostate MRI and histology via multiattribute combined mutual information

By performing registration of preoperative multiprotocol in vivo magnetic resonance (MR) images of the prostate with corresponding whole-mount histology (WMH) sections from postoperative radical prostatectomy specimens, an accurate estimate of the spatial extent of prostate cancer (CaP) on in vivo MR imaging (MRI) can be retrospectively established. This could allow for definition of quantitative image-based disease signatures and lead to development of classifiers for disease detection on multiprotocol in vivo MRI. Automated registration of MR and WMH images of the prostate is complicated by dissimilar image intensities, acquisition artifacts, and nonlinear shape differences. The authors present a method for automated elastic registration of multiprotocol in vivo MRI and WMH sections of the prostate. The method, multiattribute combined mutual information (MACMI), leverages all available multiprotocol image data to drive image registration using a multivariate formulation of mutual information. Elastic registration using the multivariate MI formulation is demonstrated for 150 corresponding sets of prostate images from 25 patient studies with T2-weighted and dynamic-contrast enhanced MRI and 85 image sets from 15 studies with an additional functional apparent diffusion coefficient MRI series. Qualitative results of MACMI evaluation via visual inspection suggest that an accurate delineation of CaP extent on MRI is obtained. Results of quantitative evaluation on 150 clinical and 20 synthetic image sets indicate improved registration accuracy using MACMI compared to conventional pairwise mutual information-based approaches. The authors' approach to the registration of in vivo multiprotocol MRI and ex vivo WMH of the prostate using MACMI is unique, in that (1) information from all available image protocols is utilized to drive the registration with histology, (2) no additional, intermediate ex vivo radiology or gross histology images need be obtained in addition to the routinely acquired in vivo MRI series, and (3) no corresponding anatomical landmarks are required to be identified manually or automatically on the images.

Determining histology-MRI slice correspondences for defining MRI-based disease signatures of prostate cancer

Mapping the spatial disease extent in a certain anatomical organ/tissue from histology images to radiological images is important in defining the disease signature in the radiological images. One such scenario is in the context of men with prostate cancer who have had pre-operative magnetic resonance imaging (MRI) before radical prostatectomy. For these cases, the prostate cancer extent from ex vivo whole-mount histology is to be mapped to in vivo MRI. The need for determining radiology-image-based disease signatures is important for (a) training radiologist residents and (b) for constructing an MRI-based computer aided diagnosis (CAD) system for disease detection in vivo. However, a prerequisite for this data mapping is the determination of slice correspondences (i.e. indices of each pair of corresponding image slices) between histological and magnetic resonance images. The explicit determination of such slice correspondences is especially indispensable when an accurate 3D reconstruction of the histological volume cannot be achieved because of (a) the limited tissue slices with unknown inter-slice spacing, and (b) obvious histological image artifacts (tissue loss or distortion). In the clinic practice, the histology-MRI slice correspondences are often determined visually by experienced radiologists and pathologists working in unison, but this procedure is laborious and time-consuming. We present an iterative method to automatically determine slice correspondence between images from histology and MRI via a group-wise comparison scheme, followed by 2D and 3D registration. The image slice correspondences obtained using our method were compared with the ground truth correspondences determined via consensus of multiple experts over a total of 23 patient studies. In most instances, the results of our method were very close to the results obtained via visual inspection by these experts.

Focal Laser Ablation for Prostate Cancer Followed by Radical Prostatectomy: Validation of Focal Therapy and Imaging Accuracy

An increased incidence of low-risk prostate cancer (PCa) has led investigators to develop focal therapy as a management option for PCa. We evaluated the effects of focal laser ablation (FLA) on PCa tissue and the accuracy of magnetic resonance imaging (MRI) in determining ablated lesion volume by comparing the whole-mount histology and MRI in four patients that underwent FLA followed by radical prostatectomy. Ablated areas were characterized by homogeneous coagulation necrosis. The MRI-calculated ablated volume correlated well with histopathology. We found that FLA creates confluent ablation with no evidence of viable cells in treated regions. Postablation MRI is able to determine the ablation accurately.

Detection of Aggressive Primary Prostate Cancer with 11C-Choline PET/CT Using Multimodality Fusion Techniques

The aim of the study was to assess whether (11)C-choline PET/CT could identify high-risk primary adenocarcinoma of the prostate. (11)C-choline PET/CT and transpelvic MRI were performed in 14 patients with untreated localized primary adenocarcinoma of the prostate, followed by radical prostatectomy as a form of primary monotherapy within 14 d of in vivo imaging. To allow accurate coregistration of whole-mount histology with in vivo imaging, additional ex vivo MR images of the prostatectomy specimen were obtained. Nonlinear 3-dimensional image deformations were used for registrations of PET/CT, MRI, and histology. Volumes of interest from tumor and benign tissue were defined on the basis of histology and were transferred into coregistered (11)C-choline PET/CT volumes to calculate the mean (T((mean))/B) and maximum (T((max))/B) ratio of tumor to benign prostate background. On the basis of MIB-1/Ki-67 expression in tumor tissues represented on a tissue microarray, we assessed whether (11)C-choline uptake correlated with local Gleason score and tumor proliferation. Histology confirmed 42 tumor nodules with Gleason scores between 3 + 2 and 4 + 4, with volumes ranging from 0.03 to 12.6 cm(3). T((mean))/B (P < 0.01) and T((max))/B (P < 0.001) ratios were significantly increased in high-Gleason score (>or=4 + 3) lesions versus 3 + 4 and lower disease but failed to distinguish between 3 + 4 disease versus 3 + 3 and lower. T((mean))/B and T((max))/B ratios were significantly increased in tumors with an MIB-1/Ki-67 labeling index greater than or equal to 5% (P < 0.01). On the basis of our preliminary data using ratios of tumor to benign prostate background, (11)C-choline preferentially identified aggressive primary prostate cancer.

Prostate cancer detection with multi‐parametric MRI: Logistic regression analysis of quantitative T2, diffusion‐weighted imaging, and dynamic contrast‐enhanced MRI

To develop a multi-parametric model suitable for prospectively identifying prostate cancer in peripheral zone (PZ) using magnetic resonance imaging (MRI). Twenty-five radical prostatectomy patients (median age, 63 years; range, 44-72 years) had T2-weighted, diffusion-weighted imaging (DWI), T2-mapping, and dynamic contrast-enhanced (DCE) MRI at 1.5 Tesla (T) with endorectal coil to yield parameters apparent diffusion coefficient (ADC), T2, volume transfer constant (K(trans)) and extravascular extracellular volume fraction (v(e)). Whole-mount histology was generated from surgical specimens and PZ tumors delineated. Thirty-eight tumor outlines, one per tumor, and pathologically normal PZ regions were transferred to MR images. Receiver operating characteristic (ROC) curves were generated using all identified normal and tumor voxels. Step-wise logistic-regression modeling was performed, testing changes in deviance for significance. Areas under the ROC curves (A(z)) were used to evaluate and compare performance. The best-performing single-parameter was ADC (mean A(z) [95% confidence interval]: A(z,ADC): 0.689 [0.675, 0.702]; A(z,T2): 0.673 [0.659, 0.687]; A(z,Ktrans): 0.592 [0.578, 0.606]; A(z,ve): 0.543 [0.528, 0.557]). The optimal multi-parametric model, LR-3p, consisted of combining ADC, T2 and K(trans). Mean A(z,LR-3p) was 0.706 [0.692, 0.719], which was significantly higher than A(z,T2), A(z,Ktrans), and A(z,ve) (P < 0.002). A(z,LR-3p) tended to be greater than A(z,ADC), however, this result was not statistically significant (P = 0.090). Using logistic regression, an objective model capable of mapping PZ tumor with reasonable performance can be constructed.

Migration of Skin Dendritic Cells in Response to Ionizing Radiation Exposure

We describe an imaging assay that monitors the migration of two unique subsets of immune dendritic cells (DC), interstitial dendritic cells (iDC) and Langerhans cells (LC), found in the dermal and epidermal layers of skin, respectively. Using this assay, we study responses of these cells to ionizing radiation. Results obtained using whole-mount histology and fluorescence microscopy suggest that ionizing radiation triggered the migration of both major histocompatibility complex (MHC) class II(+) iDC and Langerin(+) LC in a dose- and time-dependent manner. Migration appeared to be limited by local administration of recombinant IL-12, a potent immunostimulatory cytokine known to induce DNA repair. Those findings were extended to an in vivo model by injecting fluorescently conjugated anti-MHC class II antibodies intradermally into the ears of live, anesthetized mice and visualizing the DC population in the same ear before and after radiation exposure using confocal microscopy.

A Novel Approach to Energy Ablative Therapy of Small Renal Tumours: Laparoscopic High-Intensity Focused Ultrasound

ObjectiveHigh-intensity focused ultrasound (HIFU) permits targeted homogeneous ablation of tissue. The objective of this phase 1 study was to evaluate the feasibility of HIFU ablation of small renal tumours under laparoscopic control. Patients and methodsTen kidneys with solitary renal tumours were treated with a newly developed 4.0MHz laparoscopic HIFU probe. In the first two patients with 9-cm tumours, a defined marker lesion was placed prior to laparoscopic radical nephrectomy. In eight patients with a mean tumour size of 22mm (range, 11–40), the tumour was completely ablated as in curative intent, followed by laparoscopic partial nephrectomy in seven tumours. One patient had post-HIFU biopsies and was followed radiologically. Specimens were studied by detailed and whole-mount histology, including NADH stains. ResultsMean HIFU insonication time was 19min (range, 8–42), with a mean targeted volume of 10.2cm3 (range, 9–23). At histological evaluation both marker lesions showed irreversible and homogeneous thermal damage within the targeted site. Of the seven tumours treated and removed after HIFU, four showed complete ablation of the entire tumour. Two had a 1- to 3-mm rim of viable tissue immediately adjacent to where the HIFU probe was approximated, and one tumour showed a central area with about 20% vital tissue. There were no intra- or postoperative complications related to HIFU. ConclusionThe morbidity of laparoscopic partial nephrectomy mainly comes from the need to incise highly vascularized parenchyma. Targeted laparoscopic HIFU ablation may render this unnecessary, but further studies to refine the technique are needed.

Combined T2-Weighted and Diffusion-Weighted MRI for Localization of Prostate Cancer

The objective of our study was to compare T2-weighted MRI alone and T2 combined with diffusion-weighted imaging (DWI) for the localization of prostate cancer. T2-weighted imaging and DWI (b value = 600 s/mm2) were performed in 49 patients before radical prostatectomy using an endorectal coil at 1.5 T in this prospective trial. The peripheral zone of the prostate was divided into sextants and the transition zone into left and right halves. T2 images alone and then T2 images combined with apparent diffusion coefficient (ADC) maps (T2 + DWI) were scored for the likelihood of tumor and were compared with whole-mount histology results. Fixed window and level settings were used to display the ADC maps. Only tumors with an area of more than 0.13 cm2 (> 4 mm diameter) and a Gleason score of > or = 6 were considered significant. The area under the receiver operating characteristic curve (A(z)) was used to assess accuracy. In the peripheral zone, the A(z) value was significantly higher (p = 0.004) for T2 plus DWI (A(z) = 0.89) than for T2 imaging alone (A(z) = 0.81). Performance was poorer in the transition zone for both T2 plus DWI (A(z) = 0.78) and T2 (A(z) = 0.79). For the whole prostate, sensitivity was significantly higher (p < 0.001) with T2 plus DWI (81% [120/149]) than with T2 imaging alone (54% [81/149]), with T2 plus DWI showing only a slight loss in specificity compared with T2 imaging alone (84% [204/243] vs 91% [222/243], respectively). Combined T2 and DWI MRI is better than T2 imaging alone in the detection of significant cancer (Gleason score > or = 6 and diameter > 4 mm) within the peripheral zone of the prostate.

Catheter-based intraluminal optical coherence tomography (OCT) of the ureter: ex-vivo correlation with histology in porcine specimens

Intraluminal optical coherence tomography (OCT) applies coherent light to provide cross-sectional images with a spatial resolution of 10-25 microm. We compared OCT and matching whole-mount histology microscopy sections of porcine upper ureters ex vivo for visualization and delineation of different tissue layers of the ureteral wall. Porcine ureters (six specimens, 24 quadrants) were flushed with normal saline solution prior to insertion of the OCT catheter (diameter, 0.014 inch, OCT wavelength, 1,300+/-20 nm). Cross-sectional OCT images were obtained in marked locations before specimens were fixed in 4% formalin, cut at marked locations, whole-mounted, and stained with hematoxilin and eosin. Visualization and delineation of different tissue layers of the ureteral wall by OCT was compared with matching histology by two independent observers (O1,O2). OCT distinguished tissue layers of the ureteral wall in all quadrants. In OCT images, O1/O2 delineated urothelium and lamina propria in 23/24 quadrants, lamina propria and muscle layer in 19/16 quadrants, inner and outer muscle layer in 13/0 quadrants, and urothelial cell layers in 13/2 quadrants, respectively. Intraluminal OCT provides histology-like images of the ureter in porcine specimens ex vivo and reliably distinguishes between urothelium and deeper tissue layers of the ureteral wall.

The Effect of Cryosurgery and Polymethylmethacrylate in Dogs with Experimental Bone Defects Comparable to Tumor Defects

The effects of liquid nitrogen (LN) and polymethylmethacrylate (PMMA) on normal bone, bone graft incorporation, and reossification were evaluated by simulating a tumor in dogs with experimental bone cavity. Ten skeletally mature mongrel dogs (20 femora) were divided into three groups: Group I, controls; Group II, LN (with and without bone graft); and Group III, PMMA (with and without LN). Roentgenograms, whole-mount histology, and tetracycline fluorescence studies were performed on the distal femur. Correlation of these studies showed that (1) marked trabecular and bone necrosis, extending 7-12 mm around the circumference of the cavity, developed by three and seven weeks after LN but no bony necrosis occurred after PMMA; (2) the pattern of reossification following cryosurgery was delayed and abnormal, demonstrating increased calcification and metaplastic bone formation; (3) cryosurgery decreased the rate of bone graft incorporation; (4) the cryonecrotic rim following cryosurgery correlated with an abortive attempt at peripheral reossification; and (5) cryosurgery had no effect on the articular cartilage. Cryosurgery is effective in causing bone necrosis, whereas PMMA is not, and the pattern of reossification is delayed and altered by freezing. This study suggests that microvascular thrombosis with subsequent ischemic infarction of bone is a major cause of bone necrosis following cryosurgery.