GATA-3 expression is not associated with complete pathological response in triple negative breast cancer patients treated with neoadjuvant chemotherapy.

OBJECTIVE: The prognosis for patients with triple negative breast cancer (TNBC) is poor, however, a subset will demonstrate complete pathological response (pCR) to chemotherapy. GATA-3 and AR may be a negative predictors for pCR although it is unclear if these results apply to TNBC. METHODS: Patients diagnosed with TNBC and treated with neoadjuvant chemotherapy were identified. Immunohistochemistry was performed for GATA-3 and AR. Both were scored using a composite of staining intensity and percentage cells stained. The primary outcome was pCR. RESULTS: Twenty-four patients were included and 7 achieved pCR. There was no difference in the pre-chemotherapy tumor size (44±28mm vs. 54±30mm; p=0.764) or lymph node status (86% vs. 71%; p=0.629) between patients with and without pCR. GATA-3 expression was present in 20 cases (83%) while AR was present in 6 cases (25%). No AR expression was seen in 15 cases (63%) with GATA-3 positivity. There was no difference in either GATA-3 (4.3±2.7 vs. 3.6±2.5; p=0.549) or AR (1.4±2.5 vs. 1.1±2.4; p=0.778) expression between patients with and without pCR. CONCLUSIONS: GATA-3 expression is frequent in TNBC even in the absence of AR. However, neither GATA-3 nor AR are associated with pCR after neoadjuvant chemotherapy.

Fluorescence in situ hybridization as an adjunct tool in the diagnosis of primary and metastatic renal cell carcinoma in fine needle aspiration specimens.

We investigated the role of fluorescence in situ hybridization (FISH) in the diagnosis of primary renal neoplasms and lesions suspicious for metastatic renal cell carcinoma. Consecutive fine-needle aspiration biopsies (FNAB) of 39 renal masses and 41 metastatic tumours suspicious for renal cell origin were assessed with an immunohistochemical panel for CK7, RCC antigen, CD10, AMACR, PAX8, vimentin, and CD117. In addition, FISH was performed using probes for chromosomes 1p, 3p, 7, 17, X, and Y. A total of 31 of 39 primary renal masses and 33 of 41 metastatic tumors suspicious for renal origin demonstrated typical cytological and immunohistochemical (IHC) features of subtypes of renal neoplasms (40 clear cell renal cell carcinoma (RCC), 20 papillary RCC, and 4 renal oncocytomas). FISH analysis of 15 randomly selected cases each of primary and metastatic lesions revealed chromosomal abnormalities consistent with the diagnosis in 73% of these cases. Of 8 primary renal masses demonstrating atypical microscopic features and noncontributory IHC profiles, FISH was helpful in subtyping 5 (62%) of these lesions (2 clear cell RCC, 1 solid variant of oncocytic papillary RCC, 1 mixed clear cell and papillary RCC, and 1 chromophobe RCC with papillary architecture). Of 8 metastatic tumors clinically suspicious for renal cell origin and supportive, but nondiagnostic IHC, FISH revealed supportive chromosomal changes in 6 (75%) cases. In conclusion FISH analysis on FNAB material, even with limited tissue, may be contributory to the diagnosis and subtyping of RCC in diagnostically challenging biopsies.

Nuclear morphometry in flat epithelial atypia of the breast as a predictor of malignancy: a digital image-based histopathologic analysis.

OBJECTIVE: To identify morphometric features unique to flat epithelial atypia associated with cancer using digital image analysis. STUDY DESIGN: Cases with diagnosis of flat epithelial atypia were retrieved and divided into 2 groups: flat epithelial atypia associated with invasive or in situ carcinoma (n = 31) and those without malignancy (n = 27). Slides were digitally scanned. Nuclear features were analyzed on representative images at 20x magnification using digital morphometric software. RESULTS: Parameters related to nuclear shape and size (diameter, perimeter) were similar in both groups. However, cases with malignancy had significantly higher densitometric green (p = 0.02), red (p = 0.03), and grey (p = 0.02) scale levels as compared to cases without cancer. A mean grey densitometric level > 119.45 had 71% sensitivity and 70.4% specificity in detecting cases with concomitant carcinoma. CONCLUSION: Morphometry of features related to nuclear staining appears to be useful in predicting risk of concurrent malignancy in patients with flat epithelial atypia, when added to a comprehensive histopathologic evaluation.

Primary carcinoma of renal calyx.

Renal calyx carcinoma (RCXC) may mimic collecting duct carcinoma (CDC) or urothelial carcinoma (UC) of the renal pelvis. RCXC is distinguished from CDC and UC of the renal pelvis as having the tumor epicenter in the renal calyx, with limited involvement of the surrounding renal pelvis surface urothelium. In this study, we summarize our experience with this entity. Ten cases of RCXC, including 9 cases with urothelial differentiation (RCXC-UC) and 1 case with salivary gland-type differentiation (RCXC-SC), were identified. Ten consecutive cases of UC were selected for comparison, with extensive renal pelvis involvement and with secondary renal parenchymal invasion. Two cases of collecting duct carcinoma (CDC) were also examined. Immunohistochemistry (IHC) was performed on representative tissue blocks for PAX8, PAX2, CK5, CK7, CK20, p63, GATA3, AMACR, RCC, CD10, vimentin, S100, and MSA. The 10 cases of RCXC (M:F=4:6, ages: 62-91 years, mean: 76) presented with renal masses of 3-6cm. Ureteroscopic studies and renal pelvic washings showed atypical/malignant cells in three cases. Seven patients were treated with nephrectomy followed by radiation±chemotherapy, and all cases developed metastases to lymph nodes or liver/lung/bone. In all 7 cases with nephrectomy, there was extensive renal parenchymal involvement with infiltrating borders and diffuse spread along collecting ducts. Six RCXC-UC contained focal squamous differentiation. The RCXC-SC displayed features of adenoid cystic and basaloid features. In situ UC, with or without papillary components, was identified in the calyces in all 7 nephrectomy cases with remaining renal pelvis harboring small tumor burden in 5 cases, and no tumor in another 2 cases. Of the three cases without nephrectomy, no tumor in the renal pelvis could be visualized with endoscopy, however one case was associated with UC of the urinary bladder. Of 10 control UC cases, tumor was limited to the tip of renal papilla in 7 cases, extensive in 3 cases, and with no extension into the collecting ducts. RCXC-UC were all positive for p63, CK5, CK7, and PAX2, with all negative for RCC. PAX8 (70% positive) and GATA3 (50% positive) reactivity was variable. The 10 UC cases shared IHC properties with RCXC-UC but frequent negativity for PAX8 and positivity for GATA3. RCXC is an aggressive neoplasm with high risk of metastases. Similar to CDC, it is located in the renal papilla and rarely with clinically visible renal pelvis tumor or ureteral urine positive for tumor cells. Unlike CDC and non-calyceal UC, RCXC shows predominantly urothelial and squamous differentiation and is associated with an in situ component of adjacent renal calyces. By IHC, RCXC exhibited features intermediate between UC and CDC with decreased or negative immunoreactivity for PAX8 and GATA.

Extended depth of focus for transmission x-ray microscope.

A fast discrete curvelet transform based focus-stacking algorithm for extending the depth of focus of a transmission x-ray microscope (TXM) is presented. By analyzing an image stack of a sample taken in a Z-scan, a fully in-focus image can be generated by the proposed scheme. With the extended depth of focus, it is possible to obtain 3D structural information over a large volume at nanometer resolution. The focus-stacking method has been demonstrated using a dataset taken with a laboratory x-ray source based TXM system. The possibility and limitations of generalizing this method to a synchrotron based TXM are also discussed. We expect the proposed method to be of important impact in 3D x-ray microscopy.

TXM-Wizard: a program for advanced data collection and evaluation in full-field transmission X-ray microscopy.

Transmission X-ray microscopy (TXM) has been well recognized as a powerful tool for non-destructive investigation of the three-dimensional inner structure of a sample with spatial resolution down to a few tens of nanometers, especially when combined with synchrotron radiation sources. Recent developments of this technique have presented a need for new tools for both system control and data analysis. Here a software package developed in MATLAB for script command generation and analysis of TXM data is presented. The first toolkit, the script generator, allows automating complex experimental tasks which involve up to several thousand motor movements. The second package was designed to accomplish computationally intense tasks such as data processing of mosaic and mosaic tomography datasets; dual-energy contrast imaging, where data are recorded above and below a specific X-ray absorption edge; and TXM X-ray absorption near-edge structure imaging datasets. Furthermore, analytical and iterative tomography reconstruction algorithms were implemented. The compiled software package is freely available.