Impact of BRAFV600E Mutation on Event-Free Survival in Patients with Papillary Thyroid Carcinoma: A Retrospective Study in a Romanian Population.

We aimed to evaluate the prognostic value of BRAFV600E mutation in a series of 127 papillary thyroid carcinoma (PTC) cases as a single factor, and in synergic interaction with other standard risk factors. BRAFV600E mutation was assessed by real-time PCR. Event-free survival (EFS) was calculated between the date of the first evaluation and the date of occurrence of an adverse event or the date of the last known status. The prevalence of BRAFV600E mutation was 57.2%. The Kaplan-Meier analysis showed a significant reduction of EFS among cases harboring BRAFV600E mutation compared to non-mutated cases (p = 0.010). In addition, BRAFV600E mutation was found to better predict adverse outcomes when associated with the following risk factors: age ≥ 55 years old (p < 0.001), male gender (p < 0.001), conventional (p = 0.005) and tall cell (p = 0.014) histology, tumor size > 40 mm (p = 0.001), extrathyroidal extension (p = 0.001), multifocality (p = 0.001) and lymph node metastasis (p < 0.001). In univariate analysis, a 3.74-fold increased risk for a reduced EFS (p = 0.018) was found for BRAFV600E-mutated cases, but no increased risk was further confirmed by multivariate analysis. Our results highlight that BRAFV600E mutation cannot be used alone as an independent predictive factor in PTC patients, but is prognostically valuable if integrated in the context of other clinicopathological risk factors.

Micro-Fragmented Adipose Tissue as a Natural Scaffold for Targeted Drug Delivery in Brain Cancer.

Major limitations in the effective treatment of neurological cancer include systemic cytotoxicity of chemotherapy, inaccessibility, and inoperability. The capability to successfully target a drug to the tumor site(s) without incurring serious side effects-especially in the case of aggressive tumors, such as glioblastoma and neuroblastoma-would represent a significant breakthrough in therapy. Orthotopic systems, capable of storing and releasing proteins over a prolonged period at the site of a tumor, that utilize nanoparticles, liposomes, and hydrogels have been proposed. One candidate for drug delivery is Micro-Fragmented Adipose Tissue (MFAT). Easily obtained from the patient by abdominal subcutaneous liposuction (autologous), and with a high content of Mesenchymal Stem Cells (MSCs), mechanically derived nanofat is a natural tissue graft with a structural scaffold organization. It has a well-preserved stromal vascular fraction and a prolonged capacity to secrete anti-tumorigenic concentrations of pre-absorbed chemotherapeutics within extracellular vesicles. This review discusses current evidence supporting the potential of drug-modified MFAT for the treatment of neurological cancer with respect to recent preclinical and in vitro studies. Possible limitations and future perspectives are considered.

Programmed death ligand-1 (PD-L1) immunohistochemical assessment using the QR1 clone in muscle-invasive urothelial carcinomas: a comparison with reference clones 22C3 and SP263.

Programmed death ligand-1 (PD-L1) immunohistochemical (IHC) status is used to predict which patients with metastatic urothelial carcinoma (UC) will respond to immunotherapy. We aimed to compare QR1(Quartett), 22C3 (Dako), and SP263 (Ventana) detection of PD-L1 expression in muscle-invasive UCs and determine the best scoring algorithm for assessment of PD-L1 expression when using the QR1 clone. Our study included 69 UCs. For SP263 and 22C3, PD-L1-positive tumor cell (TC) and/or immune cell (IC) percentages (TC%/IC%) and the Combined Positive Score (CPS) were assessed, respectively (positivity cut-offs of ≥ 25% and ≥ 10). For QR1, both interpretation systems were evaluated. The concordances between assays were calculated. PD-L1 IHC staining characteristics were comparable between QR1, 22C3, and SP263 in both conventional and variant histology UCs. We demonstrated strong or very strong correlations between clones; the strongest correlation for TCs was between QR1 and SP263 (r = 0.92; p = 0.001) and for ICs was between QR1 and 22C3 (r = 0.85; p = 0.001). Our comparative analysis of the scoring algorithms revealed very good concordances among the three assays (range 0.791-0.878); the highest concordance was between QR1 and SP263 when CPS was used as the scoring algorithm for QR1 (0.878; p < 0.001). Our study is the first to demonstrate that the QR1 clone can be used to evaluate PD-L1 status in UCs, with a very good agreement rate with the reference clones. QR1 appeared to be more similar to the SP263 clone. With regard to the scoring algorithm, when evaluating PD-L1 expression using QR1 clone, CPS performed better compared with the TC%/IC% score.

PD-L1 Expression in Muscle Invasive Urothelial Carcinomas as Assessed via Immunohistochemistry: Correlations with Specific Clinical and Pathological Features, with Emphasis on Prognosis after Radical Cystectomy.

In the present study, we analyzed Programmed Death Ligand-1 (PD-L1) expression in radical cystectomy (RC) specimens from patients with muscle-invasive urothelial carcinoma (UC), in order to assess any correlations with specific clinicopathological features and its potential prognostic value. A multi-institutional study was performed within the departments of urology and pathology at the Mureș County Hospital, Romania, and Centre Hospitalier Lyon Sud, France. Sixty-nine patients with MIBC were included, for whom tumor histology (conventional versus histological variant/differentiation), tumor extension (T), lymph node involvement (N), and distant metastases (M) were recorded. PD-L1 immunostaining was performed using the 22C3 clone and was interpreted using the combined positive score (CPS) as recommended (Dako Agilent, Santa Clara, CA, USA). Positive PD-L1 immunostaining was more prevalent among UCs with squamous differentiation compared to conventional UCs and trended towards an improved OS (p = 0.366). We found the T stage to be a risk factor for poor survival in PD-L1-positive patients (HR 2.9, p = 0.021), along with the N stage in PD-L1-negative patients (HR 1.98, p = 0.007). No other clinicopathological factor was found to be significantly associated with PD-L1 positivity. Thus, we confirm the need for PD-L1 immunostaining prior to initiating immune checkpoint inhibitor therapy for a more accurate assessment of the patients' chances of responding to treatment.

Risk stratification of papillary thyroid microcarcinomas via an easy-to-use system based on tumor size and location: clinical and pathological correlations.

INTRODUCTION: We aimed to determine whether two clinically accessible parameters, tumor size and location within the thyroid, correlate with clinicopathological features that are predictors of high risk in papillary thyroid microcarcinomas (PTMCs). MATERIALS AND METHODS: PTMC cases were obtained from the database of the Department of Pathology, Emergency County Hospital, Târgu Mures, Romania. Four tumor groups were created based on tumor size and location: Group I (≥5 mm, subcapsular), Group II (≥5 mm, nonsubcapsular), Group III (<5 mm, subcapsular), and Group IV (<5 mm, nonsubcapsular) PTMCs. Clinicopathological features and follow-up data were compared by univariate and multivariate analysis. RESULTS: Our study included 164 PTMCs (n=70/20/19/55 in Groups I∕II∕III∕IV, respectively). High-grade morphological features, such as plump pink cells (p=0.010), tumor desmoplasia (p=0.022) and sclerosis (p=0.001), infiltrative tumor borders (p=0.005), positive resection margins (p=0.005), invasion into the perithyroid adipose tissue (p=0.001), irregular nuclear membranes (p=0.004), and pseudoinclusions (p=0.001) were significantly more prevalent among Group I PTMCs. Group IV PTMCs were characterized by a paucity of the above-mentioned morphological features, while Group II and III PTMCs displayed intermediate morphological profiles. CONCLUSIONS: Group I PTMCs proved to be associated with more aggressive morphological features and might need a more careful clinical approach.

DNA isolation from achieved formalin-fixed paraffin-embedded tissues in a series of 212 thyroid carcinoma cases: the influence of preanalytical factors on DNA quantity and purity.

OBJECTIVE: The present study aimed to investigate the influence of several important preanalytical factors (storage period of the tumor block, maximal diameter of the tumor circled area, tumor volume and tumor fraction) on the isolated DNA from formalin-fixed paraffin-embedded (FFPE) tissues in a series of thyroid carcinomas. DESIGN: Our study included 212 FFPE blocks, archived in the Department of Pathology, Târgu-Mureș Emergency County Hospital for up to 10 years. DNA isolation was performed using a commercially available kit (MasterPure DNA purification kit, Epicentre). The DNA parameters (concentration and purity) were determined using a spectrophotometer and the Qubit 2.0 Fluorometer (Thermo Fisher Scientific) for an accurate and sensitive DNA quantification. RESULTS: The mean DNA concentration and purity for the study cases were 489.3±372.6 ng/µl and 1.667±0.1912, respectively. The DNA concentration was correlated with the maximal diameter of the tumor circled area (p<0.0001), the tumor volume (p<0.0001) and tumor fraction (p=0.0462). No statistically significant differences both in terms of DNA concentration (p=0.374) and purity (p=0.125) in relation with the storage period of the tumor blocks were observed. When using a fluorometric quantification method, the DNA concentration was lower (mean DNA concentration: 47.15±32.85 ng/µl), but similar correlations with the morphological factors were observed. Apart for three cases, the real-time PCR amplification of the BRAF gene was successfully assessed in all cases. CONCLUSION: The maximal diameter of the tumor circled area, tumor volume and tumor fraction are important morphological factors that correlate with the DNA concentration and should be carefully assessed in routine practice prior to performing DNA isolation from FFPE tissues.

A 15 year institutional experience of well-differentiated follicular cell-derived thyroid carcinomas; impact of the new 2017 TNM and WHO Classifications of Tumors of Endocrine Organs on the epidemiological trends and pathological characteristics.

PURPOSE: Our study aimed to describe the evolution of the rate of pathological subtypes of well-differentiated follicular-cell derived thyroid carcinomas (DTCs) in the Department of Pathology, Emergency County Hospital Targu-Mures, Romania over a 15 year period and to assess the impact the new 2017 WHO and TNM classifications of thyroid tumors had on our cases. METHODS: The pathological data were retrieved from the original pathological reports. After applying the exclusion criteria the remaining cases were reviewed on a double-headed microscope and reclassified according to the 2017 WHO and TNM staging system. The follow-up data were collected from the Institute of Oncology Cluj-Napoca, Romania. RESULTS: Our study included 396 cases of DTCs (375 papillary, 11 follicular, and 10 Hürthle cell carcinomas). PTCs revealed a significant increasing trend over the study period, whereas follicular and Hurthle cell carcinomas remain rare; 125/131 of noninvasive encapsulated follicular variant PTC (EFVPTC) were reclassified as noninvasive follicular thyroid neoplasms with papillary-like nuclear features (NIFTPs), resulting in a 33.3% reduction in the number of PTCs. According to 2017 TNM stage-grouping 31% of 271 patients with DTC were downstaged. Follow-up data were available for most of the patients (65.7%, mean period 58.1 months). All patients with noninvasive EFVPTC were disease free at the last clinical assessment. CONCLUSIONS: The increasing rate of PTC was maintained even after exclusion of NIFTP. By applying 2017 TNM criteria, a significant number of DTC cases were downstaged into a more favorable group. Follow-up data highlight the indolent behavior of noninvasive EFVPTCs reclassified as NIFTPs.

Evaluation of a DNA Extraction and Purification Protocol Using Archived Formalin-fixed Paraffin-embedded Tissues for BRAF Mutations Analysis in Papillary Thyroid Microcarcinomas.

The isolation of good quality genomic DNA from formalin-fixed, paraffin-embedded tissues is challenging, especially in cases of small tissue specimens. The aim of our study was to evaluate a DNA extraction protocol using formalin-fixed, paraffin-embedded tissues in our laboratory and apply this method to a series of papillary thyroid microcarcinomas (PTMCs). A total of 25 PTMCs and 3 papillary thyroid carcinoma control cases were included in the study. We assessed a DNA extraction protocol on the basis of a precipitation method (MasterPure DNA purification kit, Epicentre), according to the manufacturer's instructions. All PTMCs were subject to real-time polymerase chain reaction (PCR) amplification targeting the BRAF gene and a housekeeping gene (GAPDH). BRAF gene mutations were then assessed by high-resolution melting analysis and confirmed by sequencing of the PCR products. Using this extraction method, we produced good yields of DNA (mean concentration, 147.4±77.8 ng/µL), in addition to high levels of purity (mean A260/A280 ratio: 1.63±0.1). We successfully assessed the BRAF mutation status in 24 cases (16 BRAF-negative; 8 BRAF positive), although 1 case revealed an inconclusive pattern following high-resolution melting analysis and sequencing of the PCR products. We observed no differences in the tumor size (P=0.693), storage period of the tumor block (P=0.282), DNA concentration (P=0.243), DNA purity (P=0.458), CpGAPDH (P=0.173), or CpBRAF (P=0.217) values between the BRAF-mutated and nonmutated group of PTMCs. Our findings demonstrate the importance of a reliable, reproducible DNA extraction technique for efficient PCR amplification, uniformly applied to all cases in this study, regardless of the BRAF mutation status.

Histopathological and perioperative factors influencing the length of hospital stay and patients' outcome after radical cystectomy for urothelial carcinoma.

INTRODUCTION: Length of hospital stay (LOS) is considered as a key factor in estimating outcomes after radical cystectomy (RC) in urothelial carcinoma (UC) patients. We aimed to assess whether clinical perioperative (age, gender, type of urinary diversion technique) and histopathological factors [UC variant, primary tumor, node, metastasis (pTNM) staging] could be a determining factor for LOS, as well as its influence on overall survival (OS) in a single institution, retrospective cohort study. PATIENTS, MATERIALS AND METHODS: We included a total of 69 UC patients that had RC performed in our Department during November 2011 and October 2018. Regular LOS was considered arbitrarily up to 12 days. All factors were analyzed in relation to LOS, using the chi-square and the Mann-Whitney tests. Impact of LOS on survival was assessed using the Kaplan-Meier and the Cox regression methods. RESULTS: Age was associated to increased LOS (p=0.042), as well as the type of urinary diversion (p=0.003). Patients with complex diversion were found more frequently in the prolonged LOS group (ileal conduit p=0.006, Mainz pouch p=0.15, Camey neobladder p=0.517). Histopathologically, N stage had a significant association to LOS (p=0.044). Survival analysis showed decreased survival in the prolonged LOS group (p=0.653). Cox regression found no influence of LOS (p=0.653), advanced age (p=0.518) or type of urinary diversion on OS. CONCLUSIONS: Advanced age, the complexity of urinary diversion and lymph node involvement were found as associated factors for prolonged LOS in RC patients. The impact of LOS on survival is uncertain, requiring larger, in-depth studies.

Histopathological predictive factors for the overall survival rate in patients with urothelial carcinoma of the bladder treated by radical cystectomy: a Romanian cohort study.

INTRODUCTION: Urothelial carcinoma (UC) variants are considered as having a more aggressive behavior and a more advanced stage at presentation than conventional UC. However, the evidence supporting the role of UC variants on overall survival (OS) is conflicting. We aimed to assess the impact of demographic factors (age at surgery, gender) and tumor characteristics [conventional∕variant UC, associated carcinoma in situ (CIS), associated papillary component, Tumor, Node, Metastasis (TNM) staging, positive surgical margins] on OS in a series of patients treated for UC in our Department. PATIENTS, MATERIALS AND METHODS: We performed a retrospective, cohort study and included 69 UC patients treated by radical cystectomy (RC) in our Department over an eight-year period, with complete follow-up information. Associations of UC variants as well as demographic and morphological factors with OS were assessed using univariable and multivariable Cox analysis. RESULTS: Our data showed that UC variants were statistically significantly associated with the presence of distant metastases (p=0.036) and positive surgical margins (p=0.009), but had no influence on OS (p=0.504). Further on, we demonstrated that age at surgery (p=0.045), tumor stage (p=0.012), lymph node involvement (p=0.009), and presence of positive surgical margins (p=0.002) had a statistically significant influence on OS both by univariable and multivariable Cox analysis. CONCLUSIONS: Age, tumor stage and lymph node involvement, as well as positive surgical margins represent prognostic factors in RC patients. UC variants were more likely to be associated to metastases and positive surgical margins but had no influence on OS.

Morphological features predictive for BRAF(V600E) mutation in papillary thyroid microcarcinomas.

BACKGROUND: The B-Raf proto-oncogene serine∕threonine kinase (BRAF) V600E (BRAF(V600E)) mutation represents a very specific marker for papillary thyroid carcinoma (PTC), including microcarcinomas (PTMCs). However, assessment of the BRAF(V600E) mutational status is expensive and not available in all pathology laboratories. AIM: We aimed to evaluate if we can identify those morphological features that could predict the presence of the BRAF(V600E) mutation in a series of PTMCs. MATERIALS AND METHODS: Nineteen PTMCs with analysis of 25 tumor foci were included. The following histological features were evaluated: size of the tumor, multifocality, extrathyroidal extension, tumor's border, characteristic PTC nuclear features, tumor-associated stromal reaction and histological variant. All PTMCs foci were subject to real-time polymerase chain reaction (RT-PCR) amplification targeting the BRAF gene. BRAF(V600E) mutation was assessed by high resolution melting (HRM) analysis and confirmed by Sanger sequencing. Morphological features associated with BRAF(V600E) positive and BRAF(V600E) negative PTMCs were compared using the two-tailed Fisher's exact test, with α set at ≤0.05. RESULTS: Out of the 25 PTMC foci, 16 (64%) were BRAF(V600E) negative, whereas nine (36%) were BRAF(V600E) positive. Our data showed that subcapsular localization (p=0.013), conventional histological type (p=0.05) and tumor-associated stromal reaction (moderate∕extensive fibrosis) (p=0.032) were significantly associated with the mutation. CONCLUSIONS: We have demonstrated the value of several morphological features in predicting a BRAF(V600E) mutation profile in PTMCs. All these parameters should be documented in the histopathological report, as they seem to be associated with this mutation and could serve as a risk stratification tool in the selection of patients in need for adjuvant post-surgery therapy.

A rare case of signet-ring cell carcinoma associated with poorly differentiated adenocarcinoma of the non-ampullary duodenum.

Primary duodenal cancer is a rare entity accounting for only 0.3% of all gastrointestinal cancers. Histopathologically, most duodenal cancers are mucin-producing adenocarcinomas, 34% being poorly differentiated. Signet-ring cell (SRC) carcinoma is extremely uncommon in the duodenum. Herein, we report a rare case of SRC carcinoma associated with poorly differentiated adenocarcinoma of the non-ampullary duodenum in a 74-year-old woman. The patient was admitted to the hospital for persistent epigastric pain, significant weight loss and hypochromic microcytic anemia. Esophago-gastro-duodenoscopy revealed a protruded lesion, with ulceration in the second portion of the duodenum, above the papilla. The patient was referred to surgery and pancreatico-duodenectomy with lymph node dissection was performed. The tumor consisted predominately of SRCs, Periodic Acid Schiff (PAS)-Alcian blue positive. The tumor cells were CDX2, cytokeratin (CK) 7 and CK 18/8 positive, which suggested a primary upper gastrointestinal tract site of origin. Immunostaining for mucin (MUC) 2 and MUC5AC was also positive demonstrating the duodenal goblet cells differentiation with a mixed gastric-foveolar and intestinal phenotype. Based on the morphological features and the immunohistochemical profile, a diagnosis of SRC carcinoma associated with poorly differentiated adenocarcinoma of the non-ampullary duodenum was set.

Angioinvasion as a factor for predicting aggressive outcome in primary thyroid angiosarcoma: three case reports and literature review.

We report three primary thyroid angiosarcoma (PTA) cases revealing distinctive morphological features. A systematic literature review completed our analysis to evaluate the most important morphological factors for predicting prognosis in PTAs. Three rare PTA cases were analysed. In addition, we identified 46 previously reported PTAs with available follow-up data to compare morphological features related to prognosis between patients with a favourable versus aggressive outcome. The three PTAs displayed considerable architectural heterogeneity: case 1 presented a well circumscribed tumour, extensively necrotic, with only a few highly pleomorphic vascular proliferation; cases 2 and 3 both exhibited plump epithelioid cells forming rudimentary vascular spaces or solid sheets. Case 3 also presented angioinvasion. Cases 1 and 2 were alive and disease-free at 40 and 73 months following diagnosis, respectively, whereas case 3 died within 14 months. Other significant prognostic factors were highlighted by our review and literature data analysis: increased tumour size (p = 0.042), extrathyroidal extension (p = 0.009), and distant metastases at diagnosis (p = 0.001). Although regarded as highly aggressive, PTA can also be characterised by an unusual favourable outcome. For the first time we highlight the importance of reporting angioinvasion, in cases of PTA, as a possible adverse prognostic factor.

XRCC3 Thr241Met and XPD Lys751Gln gene polymorphisms and risk of clear cell renal cell carcinoma.

INTRODUCTION: In the last decade, an increasing number of polymorphisms in DNA repair genes have been identified and their involvement in carcinogenesis was studied. Despite the fact that XRCC3 and XPD DNA repair genes association with several types of cancer was widely studied, their role in the development of clear cell renal cell carcinoma (CCRCC) has not been established in the European population. OBJECTIVE: The objective of this study was to investigate the association of XRCC3 Thr241Met and XPD Lys751Gln gene polymorphisms with the risk of CCRCC and the association between these genotypes and CCRCC histopathological prognostic factors (pathologic stage, Fuhrman grade, tumor diameter). METHODS: This study included 73 patients with CCRCC and 100 healthy individuals without cancer. We used the PCR-RFLP method to determine XRCC3 and XPD genotypes. RESULTS: The XPD 751 variant genotype (Lys/Gln) was more frequent in CCRCC patients than in healthy individuals (OR = 2.92, 95%CI: 1.47-5.79, p= 0.001). Regarding the XRCC3 Thr241Met/XPD Lys751Gln combined genotypes a significant difference was found between patients and controls for Thr/Thr+Lys/Gln (OR = 5.44, 95%CI: 2.09-14.15, p= 0.0003) and for Thr/Met+Gln/Gln (OR = 11.2, 95%CI: 1.95-100.4, p= 0.01).No association was found between any of the studied genotypes and histopathological prognostic factors of CCRCC. CONCLUSIONS: Our findings indicate that XPD Lys751Gln polymorphism may be a risk factor for CCRCC. Regarding the XRCC3 Thr241Met polymorphism, an association with CCRCC was found only in XRCC3 Thr241Met/XPD Lys751Gln combined genotypes.

The storage period of the formalin-fixed paraffin-embedded tumor blocks does not influence the concentration and purity of the isolated DNA in a series of 83 renal and thyroid carcinomas.

Optimal recovery of nucleic acids from formalin-fixed paraffin-embedded (FFPE) tissues is highly dependent on a series of pre-extraction steps, mainly related (but not limited) to fixation. The aim of our study was to investigate if the storage period of the FFPE blocks had a significant effect on the isolated DNA. We examined the quantity and purity of the isolated DNA from 83 FFPE blocks, corresponding to malignant thyroid (n=28) and renal (n=55) carcinomas that had been stored in our department for up to eight years. The DNA extraction protocol was based on a precipitation method (MasterPure™ DNA Purification Kit, Epicentre), in accordance to the manufacturer instructions, optimized in our laboratory. A spectrophotometer was used to determine the yield (A260) and purity (A260/A280 ratio) of the isolated DNA. We successfully isolated good DNA quantity and purity from all our study cases (mean concentration: 223.4 ± 104.16 ng/µL; mean A260/A280 ratio: 1.68 ± 0.09). Moreover, no statistically significant differences were observed between tumor blocks stored for 2-3 years and 7-8 years, respectively, both in terms of DNA quantity (p=0.196) and purity (p=0.663). In conclusion, we successfully validated an efficient, reproducible DNA extraction technique that provided a good range of DNA concentrations and purity, regardless the type of tissue (thyroid or kidney). Moreover, we demonstrated that the storage period of the FFPE blocks does not have a significant influence on the DNA quantity and purity.

Thyroid tumors of uncertain malignant potential: morphologic and imunohistochemical analysis of 29 cases.

INTRODUCTION: Thyroid tumors of uncertain malignant potential (TT-UMP) include follicular and well-differentiated tumors of UMP (FT-UMP/WDT-UMP), as it refers to the presence of questionable capsular/vascular invasion or incompletely developed papillary thyroid carcinoma (PTC)-type nuclear changes. However, these tumors are difficult to diagnose in most cases. We aimed to investigate whether immunohistochemistry (HBME-1, cytokeratin-19, galectin-3, CD56 and p63) provides additional information concerning such lesions. MATERIALS AND METHODS: We performed an immunohistochemical analysis on 29 TT-UMP cases (22 WDTs-UMP and 7 FTs-UMP) selected from the Rhone Alpes thyroid cancer registry and Departement of Pathology, Tîrgu-Mures Emergency County Hospital database. The clinicopathological and follow-up data were obtained. RESULTS: In the WDT-UMP group, HBME-1 was positive in 9/22 (40.9%) cases. CD56, a marker whose expression is reduced or absent in thyroid carcinoma, showed a "malignant" profile (no expression) in 13/22 (59.1%) cases. 7/22 (31.9%) cases were both HBME-1+ and CD56-. One case showed the co-expression of HBME-1, CD56, galectin-3 and cytokeratin-19. In the FT-UMP group, two cases were positive for HBME-1, other two for both galectin-3 and CK19 and only one case revealed a "malignant" CD56 profile. The follow-up data showed no distant metastases or persistent disease. CONCLUSION: Our study demonstrated very heterogeneous immunohistochemical profiles for TTs-UMP, further supporting the borderline nature of these lesions. WDTs-UMP revealed a certain tendency toward a PTC profile, suggesting a possible pathogeneticlink between these two entities. However, immunohistochemistry is still to be regarded more as a supporting diagnostic tool, while morphological criteria should always prime in the diagnostic decision.

Micropapillary urothelial carcinoma: an aggressive variant of urothelial carcinoma.

Micropapillary urothelial carcinoma (MPC) is a rare variant of urothelial carcinoma (UC) with an aggressive clinical course, an advanced stage at first presentation and a high metastatic potential. The aim or our study is to present five illustrative cases of MPC, diagnosed among the 21 patients with UC treated by radical cystectomy in the Department of Urology, County Hospital of Tirgu Mures, Romania, between January 1, 2011 and December 31, 2013. The morphological and immunohistochemical features of this rare and aggressive variant of UC, as well as a brief review of the literature are all presented. All five cases were associated with lymph node metastases with micropapillary features, regardless of the microscopic aspect of the tumor on the surgical specimens [transurethral resection (TUR) or cystectomy]. Three of them had a micropapillary component in the TUR, on the cystectomy specimen, or in both, along with lymph nodes metastases. In two cases, the MPC features were present only in the lymph node metastasis, with a conventional UC on the TUR and on the cystectomy. Immunohistochemical staining demonstrated that both micropapillary and associated conventional UC were positive for CK7 and CK20. Ki67 was expressed in 40% of tumor cells and CD34 was positive in the endothelial cells and negative in the flattened spindled cells lining the retraction spaces around tumor cell nests. MPC is a highly aggressive variant of UC with specific morphological characteristics. Any amount of micropapillary component found in UC is significant, and should be reported because it encompasses an aggressive clinical behavior and a poor prognosis.

Diagnostic value of HBME-1, CD56, Galectin-3 and Cytokeratin-19 in papillary thyroid carcinomas and thyroid tumors of uncertain malignant potential.

AIM: We aimed to evaluate four immunohistochemical markers (HBME-1, Galectin-3, Cytokeratin-19 and CD56) used alone or in panels in a series of papillary thyroid carcinoma (PTC) and thyroid tumors of uncertain malignant potential (TT-UMP) cases. MATERIALS AND METHODS: We performed an immunohistochemical analysis on a tissue micro-array of 11 PTCs [six classic (CPTC), five follicular variant (FVPTC)] and 31 TTs-UMP. A control group of 11 benign thyroid lesions/tumors was also included. RESULTS: CD56, whose expression is reduced or absent in thyroid carcinomas, was the most sensitive marker (81.8%), showing a "malignant" profile in 5/6 CPTCs and 4/5 FVPTCs. It was followed by HBME-1 (63.6% sensitivity). Cytokeratin-19 and Galectin-3 were the least sensitive antibodies (45.6%), but the most specific ones (100%). Three panels consisting of CD56 and/or Cytokeratin-19/Galectin-3 and HBME-1 and/or CD56 reached the highest sensitivity (90.9%) and the highest negative predicting value (87.5 and 83.3, respectively). In TTs-UMP, Cytokeratin-19, Galectin-3, HBME-1 and CD56 stained negatively in most of the cases (90.3%, 83.9%, 87.1% and 61%, respectively) and no statistically significant differences compared to the benign thyroid lesions' immunoprofile could be observed. CONCLUSIONS: New panels of antibodies, consisting of CD56 and/or Cytokeratin-19/Galectin-3 and CD56 and/or HBME-1 that were found to be highly sensitive for PTC in our study, are reported. Applying these panels to TTs-UMP seems also useful. Our results showed that these tumors have an immunoprofile similar to the benign thyroid lesions, suggesting that they are most likely to have a benign rather than a malignant biological behavior.

Morphologic and molecular subtype status of individual tumor foci in multiple breast carcinoma. A study of 155 cases with analysis of 463 tumor foci.

The aim of this study is to evaluate biomarker immunophenotypic heterogeneity between separate tumor foci of multiple breast carcinoma, its correlation with morphologic features and tumor grade, and its influence on the treatment. One hundred fifty-five invasive multiple breast carcinomas were retrospectively analyzed over a 6-year period (2007-2012), and the expression of estrogen (ER) and progesterone (PR) receptors, Ki-67 proliferative index, human epidermal growth factor receptor 2 expression, morphologic subtype, and tumor grading were analyzed in each tumor focus. We found mismatches in immunohistochemical features in 71 (53.78%) of 132 patients with similar histology and in 13 (56.52%) of 23 cases with different histology. When analyzing mismatches in ER and PR statuses together, in 4 (23.52%) of 17 cases, one of the tumor foci was ER or PR positive, whereas the index tumor did not express either marker. The most numerous mismatches (45 cases; 29.03%) concerned the proliferative index; in 14 cases (9.03%), the additional focus had a higher index than did the main focus, and in 9 of these cases, the additional focus displayed a histologic grade of 3. Mismatches in human epidermal growth factor receptor 2 status appeared in 25 (16.12%) cases. The histologic type of the additional foci was different from the index tumor in 23 (14.83%) cases. Assessment of all tumor foci would have determined 19 (12.25%) cases to receive different adjuvant treatments compared with what would have been indicated if only the biological status of the largest primary tumor was assessed. We strongly recommend assessing and reporting each tumor focus independently.

Immunohistochemical markers in the diagnosis of papillary thyroid carcinomas: The promising role of combined immunostaining using HBME-1 and CD56.

We aimed to evaluate the expression and diagnostic value of five immunohistochemical markers (HBME-1, Galectin-3, CK19, CD56 and p63) in a very large series of unequivocal papillary thyroid carcinoma (PTC) cases, including both the classic (CPTC) and the follicular variant (FVPTC). We performed an immunohistochemical analysis on a tissue micro-array of 204 PTCs (98 CPTCs, 90 FVPTCs, and 16 other variants). HBME-1 was the most sensitive marker, staining 95.9% of CPTCs and 81.1% of FVPTCs. CD56, a marker whose expression is reduced or absent in thyroid carcinoma, revealed a negative, "malignant" profile in 93.9% of CPTCs and 73.3% of FVPTCs. Galectin-3, CK19 and p63 were positive in 64.7%, 45.6% and 6.9% of PTCs, respectively. The immunopanel consisting of HBME-1, CD56 and/or CK19 reached the highest sensitivity (95.6%). The co-expression of 2 or more proteins was observed in 88.2% of PTCs, with HBME-1 and CD56 being the most frequent positive association (79.4%). We report a new panel of antibodies consisting of HBME-1, CK19 and CD56 that was found to be highly sensitive for both CPTC and FVPTC. This panel could be recommended as a supplement to the morphological criteria in the diagnosis of difficult FVPTC cases.