Whole-mount Sections Research Articles

Abstract 1859: Potential impact on the biology and biomarker utility of ERG-typing in the context of ethnic differences of prostate cancer

Abstract Introduction: In comparison to other ethnicities African American men have the highest incidence and mortality rates of prostate cancer (CaP). Recent reports on ethnic differences of ERG oncogenic activation, the most common CaP genomic alteration, provide new insights into the ERG based stratification of CaP in the context of the increasingly diverse ethnic patient populations in the United States. This rapidly advancing knowledge has potential to stratify CaP by ERG typing in a given patient population that may eventually lead to new targeted treatments. Methods: Whole-mounted prostate sections of 91 Caucasian American (CA) and 91 African American (AA) men matched for age, Gleason score and pathologic stage were analyzed for ERG oncoprotein expression. High grade tumors with a primary Gleason pattern of 4 or 5 or cumulative Gleason score of 8-10 from a total of 63 AA patients and 63 CA patients were also assessed for ERG. Hormone naïve AA CaP patients (n=306) with a minimum of two years of follow up after radical prostatectomy were evaluated for ERG frequencies in whole-mounted prostate specimens. Performance of PCA3, ERG and PSA biomarkers were evaluated in 306 post-DRE urine samples from CA and AA patients by the DTS® 400 system and by whole transcriptome analyses in selected CaP samples. The associations of biomarker scores with clinical parameters were assessed in matching biopsies and were stratified by ethnicity. Results: Remarkably higher frequencies of ERG expression was noted in the index tumors of CA (63.3%) compared to AA (28.6%) men. The observed difference was even more pronounced in higher grade disease AA (16%) and CA (49%), respectively. ERG positive tumors were more likely to be present in index tumors of younger AA men. Furthermore, ERG negative tumors were significantly associated with higher pathologic T stage. In urine assays the combination of ERG and PCA3 scores increased the AUC performance as compared to serum PSA alone (0.68 vs. 0.53, p = 0.0062) for predicting CaP in CA men. In contrast, the biomarker panel showed strikingly poor performance in AA men (AUC = 0.49). Conclusions: ERG-based stratification defines molecular subtypes and ethnic differences in the biology of CaP. Association of ERG negative status with high grade tumors and poor prognostic features in AA patients warrants the development of informative marker panels that includes ERG to aid in diagnostic, prognostic and targeted therapy settings. Citation Format: Albert Dobi, Yongmei Chen, Amina Ali, Denise Young, Philip Rosen, James Farrell, Michael Degon, Sudhir Srivastava, Jacob Kagan, Jocelyn Lee, Jennifer Cullen, Gyorgy Petrovics, David G. McLeod, Isabell A. Sesterhenn, Shiv Srivastava. Potential impact on the biology and biomarker utility of ERG-typing in the context of ethnic differences of prostate cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1859. doi:10.1158/1538-7445.AM2014-1859

Clinicopathological study of safe resectional margin in mid and low rectal cancer after neoadjuvant chemoradiotherapy

To investigate the regression pattern of mid and low rectal cancer treated with neoadjuvant chemoradiotherapy and then to provide the pathological proofs for reasonable resectional margin in rectal cancer surgery. Forty cases of mid and low rectal cancer patients received concurrent chemoradiotherapy and then underwent radical operation. The whole-mount serial sections of resected rectal cancer specimen were stained with cytokeratin antibody using immunohistochemical techniques to show the residual cancer cells under the mucosa. The microscopic measurement was performed to determine the reverse infiltration of cancer cells in the rectal wall and to describe the cancer cells scatter ways in the cancer mass. The Ki-67 immunohistochemical stain was also performed to show the proliferation activity of residual cancer cells after neoadjuvant chemoradiotherapy. The length of specimen was shrinking continuously during the pathologic section production and the shrink rate was 18%. There were remanent cancer cells which showed positive Ki-67 expression and the chemoradiotherapy decreased the Ki-67 expression significantly. The lower edge of remaining ulcers or scars could be used as the reference point from which the cancer infiltration could be measured. According to our measurement, the average reverse infiltration of cancer cells in the whole-mount section was (6.1±4.7) mm, the deepest one was 11.0 mm in the section which could be converted into fresh bowel length of 12.98 mm. The pathology showed that the residual cancer cells scattered in the fibrous tissue of ulcers, scars and manifested a regression of spatial distribution. The rectal cancers show regression in different degrees after neoadjuvant chemoradiotherapy. The residual cancer cells in the fiber tissues manifest proliferation activity. The distal end of resection should be at least 2 cm away from the lower edge of ulcers or scars of primary tumor in the rectal wall in patients after neoadjuvant chemoradiotherapy. The circumferential resection margin should include all the fibrous scar of the tumor area to ensure the remove of tumor cells completely.

Comparison of Real-Time Elastography and Multiparametric MRI for Prostate Cancer Detection: A Whole-Mount Step-Section Analysis

The purpose of this study was to compare prostate cancer detection rate of real-time elastography (RTE) with that of multiparametric MRI to evaluate the advantages and disadvantages of the two methods. Thirty-nine patients with biopsy-proven prostate cancer underwent both RTE and multiparametric MRI to localize prostate cancer before radical prostatectomy. RTE was performed to assess prostate tissue elasticity, and hard lesions were considered suspicious for prostate cancer. Multiparametric MRI included T2-weighted MRI, diffusion-weighted MRI (DWI), and contrast-enhanced MRI (CE-MRI) with an endorectal coil at 1.5 T. After radical prostatectomy, whole-mount step sections of the prostate were generated, and the prostate cancer detection rates with both modalities were analyzed for cancer lesions measuring 0.2 cm3 or larger. Histopathologic examination revealed 61 cancer lesions. RTE depicted 39 of 50 cancer lesions (78.0%) in the peripheral zone and 2 of 11 (18.2%) in the transitional zone. Multiparametric MRI depicted 45 of 50 cancer lesions (90.0%) in the peripheral zone and 8 of 11 (72.7%) in the transitional zone. Significant differences between the two modalities were found for the transitional zone and anterior part in prostates with volumes greater than 40 cm3 (p<0.05). Detection rates for high-risk prostate cancer (Gleason score≥4 and 3) and cancer lesions with volumes greater than 0.5 cm3 were high for both methods (93.8% and 80.5% for RTE, 87.5% and 92.7% for multiparametric MRI). Volumetric measurements of prostate cancer were more reliable with T2-weighted MRI than with RTE (Spearman rank correlation, 0.72 and 0.46). RTE and multiparametric MRI depicted high-risk prostate cancer with high sensitivity. However, multiparametric MRI seems to have advantages in tumor volume assessment and for the detection of prostate cancer in the transitional zone and anterior part within prostates larger than 40 cm3.

Accurate Determination of the Pathological Stage with Gross Dissection Protocol for Radical Cystectomy

The current protocol for reporting urinary bladder cancer in radical cystectomies may exhibit limitations in the diagnostic accuracy, such as a risk of understaging, especially in cases with prostatic involvement. Difficulty can arise in the verification of stage pT0, and the assessment of surgical margins is suboptimal. We have developed a daily gross dissection protocol practice where radical cystectomies are totally embedded and evaluated histologically in whole-mount sections. We report here on the first 138 consecutive specimens from 2008 to the first quarter of 2012 inclusive. The incidence of the cancer stages was compared with data on 15,586 radical cystectomies from the literature. The differences were analyzed with the one-sample z-test (p < 0.05). The following emerged from and our series and the literature data: pT0 8.7 % and 6.1 %; pTa 0.7 % and 2.9 %; pTis 2.9 % and 6 %; pT1 15.2 % and 15.5 %; pT2 21 % and 23.3 %; pT3 34.8 % and 34.3 %; and pT4 16.7 % and 11 %, respectively. Our findings closely reflected the means of the published statistical data based on a large number of cases. The differences were due to the more detailed processing: the case numbers in groups from pTis to pT2 were comparatively low, while those in groups pT3 and pT4 were higher. The difference in group pT4 was significant (p = 0.0494). With this method, only those samples were regarded as pT0 in which the granulomatous area and the hemosiderin deposition indicative of the earlier intervention were observable and the entire preparation was tumor-free.

Role of endorectal MR imaging and MR spectroscopic imaging in defining treatable intraprostatic tumor foci in prostate cancer: Quantitative analysis of imaging contour compared to whole-mount histopathology

PurposeTo investigate the role of endorectal MR imaging and MR spectroscopic imaging in defining the contour of treatable intraprostatic tumor foci in prostate cancer, since targeted therapy requires accurate target volume definition. Materials and methodsWe retrospectively identified 20 patients with prostate cancer who underwent endorectal MR imaging and MR spectroscopic imaging prior to radical prostatectomy and subsequent creation of detailed histopathological tumor maps from whole-mount step sections. Two experienced radiologists independently reviewed all MR images and electronically contoured all suspected treatable (⩾0.5cm3) tumor foci. Deformable co-registration in MATLAB was used to calculate the margin of error between imaging and histopathological contours at both capsular and non-capsular surfaces and the treatment margin required to ensure at least 95% tumor coverage. ResultsHistopathology showed 17 treatable tumor foci in 16 patients, of which 8 were correctly identified by both readers and an additional 2 were correctly identified by reader 2. For all correctly identified lesions, both readers accurately identified that tumor contacted the prostatic capsule, with no error in contour identification. On the non-capsular border, the median distance between the imaging and histopathological contour was 1.4mm (range, 0–12). Expanding the contour by 5mm at the non-capsular margin included 95% of tumor volume not initially covered within the MR contour. ConclusionsEndorectal MR imaging and MR spectroscopic imaging can be used to accurately contour treatable intraprostatic tumor foci; adequate tumor coverage is achieved by expanding the treatment contour at the non-capsular margin by 5mm.

Sialolipoma of Minor Salivary Gland in Uvula

Lipomatous lesions of the salivary glands are rare, accounting for less than 0.5% of all parotid gland tumours. Distinct microscopic variants of lipoma of the salivary glands, e.g. angiolipoma, fibrolipoma, pleomorphic lipoma and spindle-cell lipoma have been reported. A 45 year-old male patient with mass on uvula was presented. The specimen was capsulated, yellow coloured, soft tissue with 1.3 cm in greatest diameter. Whole-mount section showed tumor composed by mature adipocytes, salivary gland parenchymal tissue and lymphoid follicles surrounded by a fibrous capsule. Salivary gland component consist acinar and ductal elements. In some areas, glandular components were atrophic. Lymphoid follicles and focal fibrosis were seen. Oncocytic, sebaceous, and squamous metaplasia were not observed. Sialolipomas were composed predominantly of adipose tissue and showed expansive growth with fibrous capsule. Sialolipomas were reported at parotid gland, submandibular gland, hard and soft palate. To our knowledge, such a case in uvula localization was not previously presented.

Prostate Histoscanning in Clinically Localized Biopsy Proven Prostate Cancer: An Accuracy Study

To assess the accuracy of prostate histoscanning (PHS) for spatial detection and localization of prostate cancer (PCa). Prospective, single center study from January to September 2012 was conducted. Inclusion criterion was biopsy confirmed PCa in patients scheduled for radical prostatectomy. In total, 98 patients were included in the study. Results of PHS were compared against whole-mount step sectioning by the Stanford technique. A lower limit of 0.1 cm(3) was used for PHS. A dedicated 12-sector form was used for spatial correlation. The urologist and pathologist were blinded for each other's results. Sensitivity, specificity, and receiver operating characteristic curves were calculated with a logistic regression model for covariates. PHS performance for detection of PCa lesions ≥0.1 cm(3) had sensitivity of 60%, specificity of 66%, and area under the curve (AUC) of 0.63. Posterior and anterior sectors achieved sensitivity of 77%, specificity of 39%, and 28% and 84%, respectively. The model containing PHS positivity within a given sector reached sensitivity of 73.4%, specificity of 65.7%, and AUC of 0.75. In a ogistic regression model, the performance of PHS was affected by sector location, rectal distance, index, and total cancer volume (all P<0.0001) and bladder fullness (P=0.02). The best PHS accuracy was present in midposterior sectors. PHS has a potential for clinical practice, especially if PHS positivity within given sectors is taken into account. A trained operator is important. More studies are necessary to test different detection limits in various clinical settings, such as targeted biopsies and image guided focal therapy.

Tissue microarray analysis as a screening tool for neuroendocrine carcinoma of the breast

Neuroendocrine carcinoma of the breast (NCB) is a fairly recent diagnostic entity added by WHO in 2003. Since then, studies have indicated that NCB potentially displays a worse prognosis than invasive ductal carcinoma. However, due to a lack of standard use of immunohistochemical staining for neuroendocrine markers and the fact that NCB may only show slight neuroendocrine morphology that can easily be overlooked, NCB is often underdiagnosed. Consequently, there is a need for fast and reliable detection method for NCB. Here, we take a first step toward finding an easy way of identifying NCB by investigating the usefulness of tissue microarray (TMA) analysis as a screening tool. We present our findings with regard to sensitivity and specificity compared with whole-mount sections. The material consists of 240 cases of breast cancer divided into 20 TMA blocks that were all immunohistochemically stained for the neuroendocrine markers chromogranin A and synaptophysin. Cases positive in more than 50% of the tumor cells were accepted in accordance with WHO (2003) standards of NCB. Sensitivity and specificity for TMA sections vs whole-mount sections were found to be 100% and 97.8%, respectively, suggesting that TMA analysis is a reliable method for NCB detection.

Determination of an Optimal Standardized Uptake Value of Fluorodeoxyglucose for Positron Emission Tomography Imaging to Assess Pathological Volumes of Cervical Cancer: A Prospective Study

PurposeTo determine the optimal standardized uptake value (SUV) of 18F-fluorodeoxyglucose (18F-FDG) for positron emission tomography (PET) imaging, at which the PET-defined gross tumor volume (GTVPET) best matches with the pathological volume (GTVPATH) in the cervical cancer.Materials and MethodsTen patients with the cervical cancer who underwent surgery were enrolled in this study. The excised specimens were processed for whole-mount serial sections and H-E staining. The tumor borders were outlined in sections under a microscope, histopathological images were scanned and the GTVPATH calculated. The GTVPET was delineated automatically by using various percentages relative to the maximal SUV and absolute SUV. The optimal threshold SUV was further obtained as the value at which the GTVPET best matched with the GTVPATH.ResultsAn average of 85±10% shrinkage of tissue was observed after the formalin fixation. The GTVPATH was 13.38±2.80 cm3 on average. The optimal threshold on percentile SUV and absolute SUV were 40.50%±3.16% and 7.45±1.10, respectively. The correlation analysis showed that the optimal percentile SUV threshold was inversely correlated with GTVPATH (p<0.05) and tumor diameter (p<0.05). The absolute SUV was also positively correlated with SUVmax (p<0.05).ConclusionThe pathological volume could provide the more accurate tumor volume. The optimal SUV of FDG for PET imaging by use of GTVPATH as standard for cervical cancer target volume delineation was thus determined in this study, and more cases are being evaluated to substantiate this conclusion.

18F-MK-9470 PET imaging of the type 1 cannabinoid receptor in prostate carcinoma: a pilot study

BackgroundPreclinical and histological data show overexpression of the type 1 cannabinoid receptor (CB1R) in prostate carcinoma (PCa). In a prospective study, the feasibility of 18F-MK-9470 positron emission tomography (PET) imaging in patients with primary and metastatic PCa was evaluated.MethodsEight patients were included and underwent 18F-MK-9470 PET/CT imaging. For five patients with primary PCa, dynamic PET/CT imaging was performed over three acquisition intervals (0 to 30, 60 to 90 and 120 to 150 min post-injection). In malignant and benign prostate tissue regions, time activity curves of the mean standardized uptake value (SUVmean) were determined as well as the corresponding area under the curve to compare 18F-MK-9470 uptake over time. Muscle uptake of 18F-MK-9470 was used as reference for non-specific binding. Magnetic resonance imaging (MRI) was used as anatomical reference and for delineating intraprostatic tumours. Histological and immunohistochemical (IHC) examination was performed on the whole-mount histopathology sections of four patients who underwent radical prostatectomy to assess the MRI-based tumour versus benign tissue classification. For three patients with proven advanced metastatic disease, two static PET/CTs were performed 1 and 3 h post-injection. 18F-MK-9470 uptake was evaluated in bone lesions of metastatic PCa by comparing SUVmean values of metastases with these of the contralateral bone tissue.Results18F-MK-9470 uptake was significantly higher in benign and malignant prostate tissue compared to muscle, but it did not differ between both prostate tissue compartments. IHC findings of corresponding prostatic histopathological sections indicated weak CB1R expression in locally confined PCa, which was not visualized with 18F-MK-9470 PET. Metastases in the axial skeleton could not be detected while some metastases in the appendicular skeleton showed higher 18F-MK-9470 uptake as compared to the uptake in contralateral normal bone.Conclusions18F-MK-9470 PET could not detect local PCa or bone metastases in the axial skeleton but was able to visualize metastases in the appendicular skeleton. Based on these pilot observations, it seems unlikely that CB1R PET will play a significant role in the evaluation of PCa.

Comparison of imaging-based gross tumor volume and pathological volume determined by whole-mount serial sections in primary cervical cancer

ObjectiveTo investigate the accuracy of imaging-based gross tumor volume (GTV) compared with pathological volume in cervical cancer.MethodsTen patients with International Federation of Gynecology and Obstetrics stage I–II cervical cancer were eligible for investigation and underwent surgery in this study. Magnetic resonance imaging (MRI) and fluorine-18 fluorodeoxyglucose positron emission tomography (18F-FDG PET)/computed tomography (CT) scans were taken the day before surgery. The GTVs under MRI and 18F-FDG PET/CT (GTV-MRI, GTV-PET, GTV-CT) were calculated automatically by Eclipse treatment-planning systems. Specimens of excised uterine cervix and cervical cancer were consecutively sliced and divided into whole-mount serial sections. The tumor border of hematoxylin and eosin-stained sections was outlined under a microscope by an experienced pathologist. GTV through pathological image (GTV-path) was calculated with Adobe Photoshop.ResultsThe GTVs (average ± standard deviation) delineated and calculated under CT, MRI, PET, and histopathological sections were 19.41 ± 11.96 cm3, 12.66 ± 10.53 cm3, 11.07 ± 9.44 cm3, and 10.79 ± 8.71 cm3, respectively. The volume of GTV-CT or GTV-MR was bigger than GTV-path, and the difference was statistically significant (P < 0.05). No significant difference was observed between GTV-PET and GTV-path (P > 0.05). Spearman correlation analysis showed that GTV-CT, GTV-MRI, and GTV-PET were significantly correlated with GTV-path (P < 0.01). There was no significant difference in the lesion coverage factor among the three modalities.ConclusionThe present study showed that GTV defined under 40% of maximum standardized uptake value in PET images was very similar to the pathological volume of cervical cancer. This result should be replicated in a larger number of patients with cervical cancer in a future study of ours.

Does 11C-choline PET-CT contribute to multiparametric MRI for prostate cancer localisation?

The aim of this work was to determine whether 11C-choline positron emission tomography (PET)-computed tomography (CT) makes a positive contribution to multiparametric magnetic resonance imaging (MRI) for localisation of intraprostatic tumour nodules. A total of 73 patients with biopsy-proven intermediate- and high-risk prostate cancer were enrolled in a prospective imaging study consisting of T2-weighted (T2w), dynamic contrast-enhanced (DCE) and diffusion-weighted (DW) MRI and 11C-choline PET-CT before radical prostatectomy. Cancerous regions were delineated on the whole-mount prostatectomy sections and on the different MRI modalities and analysed in 24 segments per patient (3 sections, 8 segments each). To analyse PET-CT images, standardized uptake values (SUV) were calculated per segment. In total, 1,752 segments were analyzed of which 708 (40.4%) were found to be malignant. A high specificity (94.7, 93.6 and 92.2%) but relatively low sensitivity (31.2, 24.9 and 44.1%) for tumour localisation was obtained with T2w, DCE and DW MRI, respectively. Sensitivity values significantly increased when combining all MRI modalities (57.2%). For PET-CT, mean SUVmax of malignant octants was significantly higher than mean SUVmax of benign octants (3.68±1.30 vs. 3.12±1.02, p<0.0001). In terms of accuracy, the benefit of adding PET-CT to (multiparametric) MRI was less than 1%. The additional value of 11C-choline PET-CT to MRI in localising intraprostatic tumour nodules is limited, especially when multiparametric MRI is used.

Dendritic and lymphocytic cell infiltration in prostate carcinoma.

We examined the distribution of CD1a⁺ cells and CD8⁺ and CD4⁺ T lymphocytes in prostate cancer (PCa) and correlated these with clinicopathological parameters. We also investigated whether the distribution of these cells was related to the expression of the cell membrane protein B7-H3, a putative negative regulator of the immune response expressed on PCa cells. A cohort of 151 PCa patients treated with radical prostatectomy (RP) was followed prospectively from 1985 until 2006 with a median follow-up of 9 years. Whole-mount sections of PCa specimens were immunostained to identify immune cells. A low number of CD1a⁺ cells was significantly associated with a high Gleason score and high pathological stage of pT3. The number of CD1a⁺ cells correlated significantly with the number of intratumoral and stromal CD8⁺ and stromal CD4⁺ lymphocytes. Kaplan-Meier analysis showed a tendency toward impaired biochemical progression-free survival in patients with few CD1a⁺ cells within their RP specimens. The expression of B7-H3 correlated inversely with the number of CD1a⁺ cells and intratumoral CD4⁺ lymphocytes; there was a trend for a similar inverse relationship between B7-H3 expression and the number of CD8⁺ lymphocytes.

Why prostate tumour delineation based on apparent diffusion coefficient is challenging: An exploration of the tissue microanatomy

Background. Focal boosting of prostate tumours to improve outcome, requires accurate tumour delineation. For this, the apparent diffusion coefficient (ADC) derived from diffusion-weighted MR imaging (DWI) seems a useful tool. On voxel level, the relationship between ADC and histological presence of tumour is, however, ambiguous. Therefore, in this study the relationship between ADC and histological variables was investigated on voxel level to understand the strengths and limitations of DWI for prostate tumour delineation. Material and methods. Twelve radical prostatectomy patients underwent a pre-operative 3.0T DWI exam and the ADC was calculated. From whole-mount histological sections cell density and glandular area were retrieved for every voxel. The distribution of all variables was described for tumour, peripheral zone (PZ) and central gland (CG) on regional and voxel level. Correlations between variables and differences between regions were calculated. Results. Large heterogeneity of ADC on voxel level was observed within tumours, between tumours and between patients. This heterogeneity was reflected by the distribution of cell density and glandular area. On regional level, tumour was different from PZ having higher cell density (p = 0.007), less glandular area (p = 0.017) and lower ADCs (p = 0.017). ADC was correlated with glandular area (r = 0.402) and tumour volume (r = −0.608), but not with Gleason score. ADC tended to decrease with increasing cell density (r = −0.327, p = 0.073). On voxel level, correlations between ADC and histological variables varied among patients, for cell density ranging from r = −0.439 to r = 0.261 and for glandular area from r = 0.593 to r = 0.207. Conclusions. The variation in ADC can to a certain extent be explained by the variation in cell density and glandular area. The ADC is highly heterogeneous, which reflects the heterogeneity of malignant and benign prostate tissue. This heterogeneity might however obscure small tumours or parts of tumours. Therefore, DWI has to be used in the context of multiparametric MRI.

Expression of hypoxia-inducible factor-1α and -2α in whole-mount prostate histology: Relation with dynamic contrast-enhanced MRI and Gleason score

The aim of this study was to investigate the association between the immunohistochemical expression of hypoxia-inducible factor (HIF)-1α and HIF-2α and dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) parameters Ktrans and kep in prostate cancer. Therefore, 15 patients with biopsy-confirmed prostate cancer underwent a pre-operative 3T DCE-MRI scan. Immunohistochemical analysis of HIF-1α and HIF-2α, and of CD31 for microvessel density (MVD) was performed. Tumor areas were delineated on whole-mount histopathological sections. Nuclear HIF expression was correlated with the quantitative DCE-MRI parameters Ktrans and kep, MVD and Gleason score. HIF expression was highly heterogeneous within tumors and between patients. Pronounced expression of HIF-2α was present, while HIF-1α expression was more limited. Larger tumors showed higher HIF-2α expression (p=0.041). A correlation between HIF-2α and Ktrans p5th was found (r=0.30, p=0.02), but no differences in Ktrans, kep and MVD were observed for different levels of HIF expression. HIF expression was not associated with Gleason score. In conclusion, in this whole-mount prostate cancer study, larger prostate tumors showed frequently high HIF-2α expression, suggesting that larger tumors are clinically most relevant. However, HIF-1α and HIF-2α were not correlated with DCE-MRI parameters. Given the pronounced expression of HIF-2α and independence of Gleason score, HIF expression may function as a biomarker to guide boost dose prescription.

EWSR1 Genetic Rearrangements in Salivary Gland Tumors

The Ewing sarcoma breakpoint region 1 (EWSR1) is translocated in many sarcomas. Recently, its rearrangement has been described in salivary gland hyalinizing clear cell carcinomas (HCCCs) and in a subset of soft tissue myoepitheliomas. This study examines the presence of the EWSR1 rearrangement in a variety of salivary gland lesions including classic myoepitheliomas and HCCCs. Using a tissue microarray and whole-mount sections, fluorescence in situ hybridization (FISH) was performed on a variety of salivary gland lesions including HCCCs. The EWSR1 rearrangement was detected in 87% of HCCCs (13 of 15); all other salivary gland lesions including classic myoepitheliomas had intact EWSR1. Patients with HCCCs with rearranged EWSR1 included 1 man, 10 women, and 2 of unknown sex. Ages ranged from 35 to 83 years; the tumor size ranged from 0.8 to 5.5 cm, and the involved locations included: palate (2), base of the tongue (2), mandible (2), submandibular gland (2), lip (1), floor of the mouth (1), sublingual gland (1), inner cheek (1), and nasopharynx (1). All HCCCs were composed of sheets and nests of monotonous cells with clear cytoplasm within a hyalinized stroma. All tested cases were immunoreactive with antibodies to p63 and were nonreactive with antibodies to more conventional myoepithelial antigens (e.g., smooth muscle actin and S100 protein). These findings show that the EWSR1 rearrangement is almost a defining feature of HCCCs and also confirm that classic salivary gland myoepitheliomas are distinct from these tumors and do not share a pathogenetic relationship with their soft tissue counterparts.

Influence of imaging and histological factors on prostate cancer detection and localisation on multiparametric MRI: a prospective study

To assess factors influencing prostate cancer detection on multiparametric (T2-weighted, diffusion-weighted, and dynamic contrast-enhanced) MRI. One hundred and seventy-five patients who underwent radical prostatectomy were included. Pre-operative MRI performed at 1.5 T (n = 71) or 3 T (n = 104), with (n = 58) or without (n = 117) an endorectal coil were independently interpreted by two radiologists. A five-point subjective suspicion score (SSS) was assigned to all focal abnormalities (FAs). MR findings were then compared with whole-mount sections. Readers identified 192-214/362 cancers, with 130-155 false positives. Detection rates for tumours of <0.5 cc (cm(3)), 0.5-2 cc and >2 cc were 33-45/155 (21-29 %), 15-19/35 (43-54 %) and 8-9/12 (67-75 %) for Gleason ≤6, 17/27 (63 %), 42-45/51 (82-88 %) and 34/35 (97 %) for Gleason 7 and 4/5 (80 %), 13/14 (93 %) and 28/28 (100 %) for Gleason ≥8 cancers respectively. At multivariate analysis, detection rates were influenced by tumour Gleason score, histological volume, histological architecture and location (P < 0.0001), but neither by field strength nor coils used for imaging. The SSS was a significant predictor of both malignancy of FAs (P < 0.005) and aggressiveness of tumours (P < 0.00001). Detection rates were significantly influenced by tumour characteristics, but neither by field strength nor coils used for imaging. The SSS significantly stratified the risk of malignancy of FAs and aggressiveness of detected tumours. • Prostate cancer volume, Gleason score, architecture and location are MRI predictors of detection. • Field strength and coils used do not influence the tumour detection rate. • Multiparametric MRI is accurate for detecting aggressive tumours. • A subjective suspicion score can stratify the risk of malignancy and tumour aggressiveness.

Postnatal development of cerebellar zones revealed by neurofilament heavy chain protein expression

The cerebellum is organized into parasagittal zones that control sensory-motor behavior. Although the architecture of adult zones is well understood, very little is known about how zones emerge during development. Understanding the process of zone formation is an essential step toward unraveling how circuits are constructed to support specific behaviors. Therefore, we focused this study on postnatal development to determine the spatial and temporal changes that establish zonal patterns during circuit formation. We used a combination of wholemount and tissue section immunohistochemistry in mice to show that the cytoskeletal protein neurofilament heavy chain (NFH) is a robust marker for postnatal cerebellar zonal patterning. The patterned expression of NFH is initiated shortly after birth, and compared to the domains of several known zonal markers such as zebrin II, HSP25, neurogranin, and phospholipase Cβ4 (PLCβ4), NFH does not exhibit transient expression patterns that are typically remodeled between stages, and the adult zones do not emerge after a period of uniform expression in all lobules. Instead, we found that throughout postnatal development NFH gradually reveals distinct zones in each cerebellar lobule. The boundaries of individual NFH zones sharpen over time, as zones are refined during the second and third weeks after birth. Double labeling with neurogranin and PLCβ4 further revealed that although the postnatal expression of NFH is spatially and temporally unique, its pattern of zones respects a fundamental and well-known molecular topography in the cerebellum. The dynamics of NFH expression support the hypothesis that adult circuits are derived from an embryonic map that is refined into zones during the first 3-weeks of life.

Prostate Cancer: All Aspects

Prostate Cancer: All Aspects

Increased rate of positive biopsies using a combination of MR-Tomography, spectroscopy and diffusion-weighted magnetic resonance imaging prior to prostate biopsies in patients with persistent elevated prostate-specific antigen values: A retrospective analysis

Purpose:Persistently elevated prostate-specific antigen (PSA) values following negative biopsies result in a diagnostic dilemma. In order to improve detection rates in patients with former negative biopsies and persistently elevated PSA values, magnetic resonance tomography (MRT), magnetic resonance spectroscopy (MRS), and diffusion-weighted magnetic resonance imaging (DW-MRI) were performed prior to prostate rebiopsies.Materials and Methods:Over a 14-month period, 67 patients (mean age of 66 years) with a history of 1-5 negative biopsies underwent endorectal magnetic resonance imaging (MRI) using T2-weighted MRT MRS and DW-MRI before an additional prostate biopsy was performed. Subsequently, 5 contrast-enhanced transrectal ultrasound-guided biopsies were performed according to a 10-core systematic scheme. Out of the 67 men, 25 patients had positive biopsies and opted for radical prostatectomy. Histological evaluation of cancer localization, PSA, diameters of primary tumors, numbers and diameters of satellite tumors, prostate volume, and staging pathology was performed. These findings were compared with MRI and MRS results.Results:Serum PSA levels ranged from 3.1 to 19.5 g/ml (median level of 7.96 ng/ml). After the 25 patients underwent radical prostatectomy, analysis of 20 whole-mount sections of 25 radical retropubic prostatectomy (RPE) specimens presented results agreeing with the tumor location from MRI and MRS data.Conclusions:The aim of image-guided diagnostics should be to provide more critical information prior to biopsy. Furthermore, the acquisition of such data is important for better risk stratification in therapeutic decisions.