Expression of E-Cadherin and Ber-EP4 in the Trophoblastic Tissues of Intrauterine and Ectopic Tubal Pregnancies.

Introduction: We investigated the role of E-cadherin and Ber-EP4 in tubal pregnancy by comparing their expressions in epithelial and trophoblastic cells both in ectopic tubal and intrauterine pregnancies. Methods: The Formalin-fixed paraffin embedded blocks of 17 intrauterine and 17 tubal pregnancies were immunohistochemically stained with E-cadherin and Ber-EP4. Results: E-cadherin was expressed in the epithelium, villous and extravillous trophoblast in tubal and intrauterine pregnancies but not in the syncytiotrophoblast. The staining intensity was lower in the extra-villous trophoblast in tubal ectopic pregnancies compared with intrauterine pregnancies. Ber-EP4 was expressed in the epithelium of tubal and intrauterine pregnancies and only in villous cytotrophoblast. The intensity of staining in tubal pregnancy was higher than in intrauterine pregnancy. Discussion: The loss of E-cadherin expression in extra-villous trophoblast and increased expression of Ber-EP4 in the villous cytotrophoblast may play a role in the formation of tubal pregnancy by allowing the blastocyst to attach to the tubal epithelium.

Ber-EP4 staining patterns on basal cell carcinomas.

OBJECTIVE: This article aimed to study two different parameters of basal cell carcinoma (BCC): First, to analyze the expression of antihuman epithelial antigen (Ber-EP4) on the primary and recurrent BCCs on the head, neck, and other body parts and second, to find Ber-EP4's staining pattern and staining intensities correlation between histological type, demographic data, tumor, and its prognostic parameters. METHODS: We evaluated the Ber-EP4 staining patterns of 201 patients diagnosed with BCC. We analyzed the possible correlation between the tumor's prognostic parameters and the Ber-EP4 staining intensity and its pattern (peripheral, superficial, or diffused). RESULTS: In 199 out of the 201 cases, staining was observed. Two cases were unstained. In 25.6% (n=51) of the cases with staining, the staining was weak, on the 25.6% (n=51), it was moderate, and on the 48.8% (n=97), it was severe. The staining pattern was 31.2% (n=62) peripheral, 4.0% (n=8) superficial, 54.7% (n=109) diffuse, and 10.1% (n=20) peripheral and superficial. CONCLUSION: Ber-EP4 is the only antibody commonly used for BCC diagnosis; the existence of different staining intensities and patterns in BCC tumor cells in routine dermatopathology practice limit the pathologists. The studies investigating Ber-EP4 staining in BCCs were conducted with very small numbers of cases. In these studies, even the presence of staining in the focal area was considered to be a positive acceptance criterion; the staining intensity and pattern were not evaluated. Therefore, our study is the first study with a high number of cases and the first to include an evaluation of Ber-EP4 staining's intensity and localization.

The utility of claudin-4 versus MOC-31 and Ber-EP4 in the diagnosis of metastatic carcinoma in cytology specimens.

BACKGROUND: Claudin-4 is a sensitive and specific marker for carcinoma in effusion cytology. The authors examined the diagnostic use of claudin-4 versus MOC-31 and Ber-EP4 by comparing their sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) in differentiating carcinoma from mesothelioma and benign/mesothelial hyperplasia in effusion specimens. METHODS: This retrospective study comprised a cohort of 229 cytology specimens, including 211 effusion fluid and 18 fine-needle aspiration specimens. Cytologic categories included 134 carcinoma, 28 mesothelioma, 46 indefinite (suspicious and atypical), and 21 benign. Cell block sections were stained for claudin-4 and compared with those previously stained for MOC-31 and Ber-EP4. Indefinite cases were further reclassified based on clinical and pathologic findings into benign (26 cases), mesothelioma (11 cases), and carcinoma (nine cases). RESULTS: None of the mesotheliomas (0/39) or benign effusions (0/47) were positive for claudin-4, whereas 134 of the 143 carcinoma specimens were positive. Compared to MOC-31 and Ber-EP4, claudin-4 had the highest specificity and PPV (100% for each), followed by Ber-EP4. Claudin-4 showed high sensitivity (93.7%), albeit lower than MOC-31. MOC-31 had the lowest specificity and PPV but the highest sensitivity and NPV. Ber-EP4 had the lowest sensitivity (91.6%). CONCLUSIONS: Claudin-4 can be used as a single marker for carcinoma with high sensitivity and superior specificity compared with MOC-31 and Ber-EP4. Mesothelial lineage can be ruled out when claudin-4 is positive. In equivocal cytology samples with few scattered cells of interest, a panel of claudin-4 and Ber-EP4 results in the highest combined sensitivity and specificity.

Differences in actin expression between primary and recurrent facial basal cell carcinomas as a prognostic factor of local recurrence.

INTRODUCTION: Due to a relatively high recurrence rate of facial basal cell carcinoma (BCC), its morbidity is very significant. AIM: To analyse the expression of α-SMA, E-cadherin, Ber-Ep4 and MOC-31 as predictors of local recurrence in a group of patients with primary and recurrent BCCs of the face in correlation with histological and clinical data. MATERIAL AND METHODS: The study cohort included 79 patients with facial BCC (52 with primary BCC and 27 with recurrent BCC) who were treated at the Department of Cranio-Maxillofacial Surgery of the Jagiellonian University, Krakow in 1997-2009. RESULTS: Significant risk factors for local recurrence included: recurrent tumour (p = 0.001), multifocal BCC (p = 0.01), incomplete tumour excision (p = 0.02) and the aggressive infiltrating histologic subtype of BCC (p = 0.05). In the group of primary BCCs, positive expression of stromal α-SMA (p = 0.03) correlated with a statistically significant higher recurrence rate and so did positive expression of α-SMA in tumour cells of recurrent BCC (p = 0.002). In the group of primary aggressive BCC subtypes, reduced expression of MOC-31 was also associated with a higher rate of relapse (p = 0.02). CONCLUSIONS: Our findings provide information about α-SMA and MOC-31 expression in primary and recurrent BCCs. These data may contribute to the formulation of a more targeted treatment plan and follow-up strategy for patients with facial basal cell carcinoma.

Ber-EP4 staining in effusion cytology: A potential source of false positives.

The distinction between reactive mesothelium and carcinoma in serous effusions can be very difficult. Immunocytochemistry (ICC) is the most widely used tool to improve the diagnostic accuracy of body fluid cytology, with several ICC markers being proposed. Ber-EP4 antibody has shown high sensitivity and specificity rates for diagnosing metastatic carcinoma. In our department, we have detected Ber-EP4 positivity in mesothelium in some cytological specimens. We reviewed all articles on Ber-EP4 staining in effusion cytology, summarized current findings and analyzed the staining pattern of all cases expressing Ber-EP4. Some cases showing Ber-EP4 positivity in mesothelium have been reported, most of which showed only weak Ber-EP4 staining or staining of less than 50% of mesothelial cells. However, some cases may show strong positivity both in cytological and histological specimens. Clinicians and pathologists should be aware of this source of misdiagnosis, and ICC results in mesothelium should be always interpreted cautiously and correlated with clinical tests, other ICC markers and patient's previous history.

Claudin-4 shows superior specificity for mesothelioma vs non-small-cell lung carcinoma compared with MOC-31 and Ber-EP4.

The distinction of malignant mesothelioma from non-small-cell lung carcinoma (NSCLC) usually requires immunohistochemistry, but some broad-spectrum carcinoma markers stain mesotheliomas, and it remains unclear which broad-spectrum markers are most valuable for distinguishing these malignancies. Here, we directly compared the sensitivity and specificity of three broad-spectrum carcinoma markers, claudin-4, Ber-EP4, and MOC-31, for distinguishing NSCLC from mesothelioma. Immunohistochemistry was performed on tissue microarrays containing 68 epithelioid mesotheliomas, 31 sarcomatoid mesotheliomas, and 147 non-small-cell lung cancers (53 adenocarcinomas, 60 squamous cell carcinomas, 13 large-cell carcinomas, and 21 sarcomatoid carcinomas). For adenocarcinoma, squamous cell carcinoma, and large-cell carcinoma, claudin-4 staining was present in 103 of 126 cases (82%), MOC-31 staining was present in 112 of 126 cases (89%), and Ber-EP4 staining was present in 113 of 126 cases (90%); these values were not statistically different. Claudin-4 stained 0 of 68 (0%), MOC-31 stained 22 of 68 (32%), and Ber-EP4 stained 24 of 68 (35%) epithelioid mesotheliomas; thus, the specificities for NSCLC versus epithelioid mesothelioma were 100%, 68%, and 65%, respectively. Claudin-4 staining was present in 7 of 21 (33%), MOC-31 staining was present in 8 of 21 (38%), and Ber-EP4 staining was present in 5 of 21 (24%) sarcomatoid carcinomas. All three markers were negative in 12 of 21 (57%) sarcomatoid carcinomas. Sarcomatoid mesotheliomas were not stained with any of these markers. We conclude that claudin-4 has considerably greater specificity and comparable sensitivity to MOC-31 and Ber-EP4 for distinguishing NSCLC from epithelioid malignant mesothelioma. The use of all three markers may be necessary for sarcomatoid neoplasms, given their limited sensitivity.

Reactivity with the EpCAM-specific antibodies MOC-31 and Ber-Ep4 in plasma cell neoplasms: a potential diagnostic pitfall in cytology samples.

OBJECTIVE: Epithelial cell adhesion molecule (EpCAM) is a protein expressed on surfaces of healthy epithelia, and is overexpressed in dysplasias and carcinomas. Immunohistochemistry (IHC) utilizing antibodies that react with EpCAM, such as MOC-31 and Ber-EP4, distinguish reactive mesothelial cells from carcinomas in serous effusions. IHC is crucial in effusions with singly dispersed atypical cells, a scenario with a broad differential, including hematopoietic malignancies. Plasma cell neoplasms (PCN) are the second most common hematopoietic malignancy, manifesting as multiple myeloma or plasmacytoma, with 6% of cases developing serous cavity involvement. Most PCNs are readily recognizable; however, variants that deviate from the classic cytomorphology risk erroneous diagnosis. This study demonstrates EpCAM expression in a subset of PCNs, highlighting a potential diagnostic pitfall in serous effusion cytology. METHODS: A 10-year retrospective search for cytology specimens with a diagnosis of PCN was performed. All cases demonstrating CD138/CD38 and monoclonal immunoglobulin expression, and adequately cellular cell block were included. IHC analysis for MOC-31 and Ber-EP4 was performed using Ventana Benchmark Ultra. Scoring was performed as follows: total IHC score equals the positive proportion (0 = no positive tumor cells; 1 = <1%; 2 = 1-10%; 3 =11-33%; 4 = 34-66%; 5 = 67-100%) plus staining intensity (0, no staining; 1, weak; 2, moderate; 3, strong). A score > 4 was considered positive. RESULTS: 2 of 28 (7%) PCNs demonstrated positivity for MOC-31 and Ber-Ep4. CONCLUSION: A subset of PCNs in cytology samples show positivity for MOC-31 and Ber-EP4 which could result in misinterpretation as carcinoma.

Expression of EpCAM in adenoid cystic carcinoma.

The mutational landscape of adenoid cystic carcinoma (ACC) is currently being revealed, but further studies are needed to identify biomarkers as therapeutic targets or prognostic factors of ACC. In this study, we investigated the expression of epithelial cell adhesion molecule (EpCAM) in ACCs. We retrospectively collected 83 cases of surgically resected ACCs. Using tissue microarray, we conducted immunohistochemical staining using the anti-EpCAM antibody. EpCAM expression was analysed by intensity score and the total immunostaining score. The positivity was 97.6% (81/83 cases), regardless of the intensity score. A higher histological grade (p = 0.006) and specific tumour location (non-salivary gland origin, p = 0.02) showed a correlation with higher EpCAM intensity. Higher EpCAM expression by total immunostaining score was associated with histological grade (p = 0.004), distant metastasis (p = 0.004) and poorer prognosis (overall survival p = 0.015 and progression-free survival p = 0.033). We suggest EpCAM as a candidate prognostic marker and a putative therapeutic target in ACC. Also, ACCs arising from salivary gland and non-salivary gland sites, respectively, might display different pathophysiologies in which EpCAM could play a role.

Granular cell basal cell carcinoma: A case report.

Granular cell basal cell carcinoma (BCC) is a rare histopathological variant of BCC. Our review of the literature revealed only 17 previously identified cases. We report the case of a 47-year-old man who presented with an ulceration on his right upper lip which was subsequently removed. Histopathologic examination revealed that the tumor was composed solely of granular cells with numerous cytoplasmic eosinophilic round inclusion bodies. Mitotic figures ranged from 8 to 15 per 10 high-power fields, with a Ki-67 proliferative index of ~5%. Immunohistochemically, the granular cells showed strong and diffuse positivity for Ber-EP4, pan-cytokeratin, AE1/AE3, CK5/6 and p63 and focal positivity for lysozyme, CD68 (clone KP1) and Bcl-2.

[The fluorescent immune cytochemical analysis of exudations and washouts from serous cavities under intra-operational cytological diagnostic.]

According publications' data, the range of diagnostic sensitivity of cytological analysis of exudates makes up to 64%-96%. In case of emergency intra-operational diagnostic, a necessity occurs to implement additional techniques. One of them is a fluorescent immune cytochemical analysis with monoclonal antibodies to BER-EP4, conjugated with fluorochrome FITC. The mentioned analysis demonstrates high diagnostic specificity and sensitivity under differential diagnostic of metastatic and reactive exudates.

Early Detection of Desiccation and Curettage Failure in the Treatment of Basal Cell Carcinoma.

BACKGROUND/AIMS: Basal cell carcinoma (BCC) is a malignant neoplasm of keratinocytes. Electrodessication and curettage (ED&C) published cure rates vary widely, and the authors of this study are unaware of any previous literature which has attempted to rapidly identify treatment failures. OBJECTIVE: To identify BCC ED&C failures by histologically analyzing the fragments produced by the third round of curettage. METHODS: The monitoring of routine therapy of 862 cases of BCC that were treated by ED&C followed by the submission of cautery fragments of the third round of curettage for histological and immunohistochemical testing. RESULTS: Of the 862 cases, 764 (89%) had no residual BCC seen in their curetting. Of these patients, zero recurrences (0%) were noted. Forty-eight of the 862 cases had residual BCC seen in their curetting and elected to receive no additional therapy. Eighteen (38%) had a recurrence detected. Fifty of the 862 cases had residual BCC seen in their curetting and elected for immediate re-excision. Thirty-five (70%) had histological evidence of residual BCC. LIMITATIONS: The study was performed at a single center with 2 years of follow-up. CONCLUSION: Pathological examination of curettage fragments in combination with immunohistochemistry testing appears to be beneficial in predicting which patients are likely to have recurrence of BCC after ED&C.

Immunocytochemical utility of claudin-4 versus those of Ber-EP4 and MOC-31 in effusion cytology.

BACKGROUND: Recently, claudin-4 (CL4) immunocytochemistry was reported to be useful for differential diagnosis in effusion cytology. We wondered whether CL4 might be useful for "single-shot" identification of metastatic carcinoma. The purpose of this study was to evaluate the usefulness of CL4 in effusion cytology. METHODS: In total, 266 cases (169 metastatic carcinomas, eight malignant mesotheliomas, and 89 reactive mesothelial cells) were selected. Immunocytochemical examinations of cell-block sections were performed for CL4, Ber-EP4, and MOC-31. We used an arbitrary 4-tiered scale based on both staining intensity and positive-cell percentage among all target cells, and calculated a staining index score (sum of the above two scores). RESULTS: In a ROC-curve analysis, higher area-under-curve values were found for CL4 than for Ber-EP4 or MOC-31 (0.982, 0.942, and 0.926, respectively). CONCLUSIONS: Since CL4 exhibited similar or superior usefulness to Ber-EP4 and MOC-31, it could become the first choice for the above differential diagnosis in effusion cytology. Diagn. Cytopathol. 2016;44:499-504. © 2016 Wiley Periodicals, Inc.

Basal cell carcinoma vs basaloid squamous cell carcinoma of the skin: an immunohistochemical reappraisal.

Typical cutaneous basal cell carcinoma (BCC) and squamous cell carcinoma (SCC) are morphologically dissimilar. It is well known, however, that poorly differentiated SCC may assume a basaloid phenotype, complicating the histologic distinction between these 2 neoplasms. Selected immunohistochemical stains have been used in the past to aid in that differential diagnosis. In the current study, additional markers were evaluated to determine whether they would be helpful in that regard. Twenty-nine cases of metatypical (squamoid) BCC (MBCC) and 25 examples of basaloid SCC (BSCC) were studied using the antibodies Ber-EP4 and MOC-31 as well as a plant lectin preparation from Ulex europaeus I (UEA-1). The resulting immunostains were interpreted independently by 3 pathologists, and the results showed that MBCCs demonstrated strong and diffuse staining for Ber-EP4 (25/29) and MOC-31 (29/29). In contrast, BSCCs tended to be only sporadically reactive for both markers (4/25 and 1/25 cases, respectively). Labeling for UEA-1 was observed in almost all BSCCs (24/25), but only 6 of 29 cases of MBCC showed limited, focal staining with that lectin. These data suggest that MOC-31 is a useful marker in the specified differential diagnosis, especially when used together with UEA-1.

Claudin-4 as a marker for distinguishing malignant mesothelioma from lung carcinoma and serous adenocarcinoma.

We compared claudin-4 with Ber-EP4 and carcinoembryonic antigen as markers to distinguish mesothelioma from lung adenocarcinoma, poorly differentiated lung squamous cell carcinoma, and serous adenocarcinoma of the uterus or ovary. All mesothelioma specimens were negative for claudin-4, but 3 of 18 specimens were focally positive for Ber-EP4. In contrast, lung adenocarcinoma including poorly differentiated adenocarcinoma was highly positive for claudin-4, but expression of Ber-EP4 and carcinoembryonic antigen varied widely. Claudin-4 in poorly differentiated squamous cell carcinoma had a lower positive expression rate than in adenocarcinoma. Granular claudin-4 immunoreactivity was conspicuous in poorly differentiated squamous cell carcinoma; this immunoreactive pattern was also observed in mesothelioma. Claudin-4 was thus considered very useful marker for distinguishing mesothelioma and adenocarcinoma, even if histological specimens are small, as in biopsies that contain limited numbers of tumor cells. However, it should be mentioned that claudin-4 has a limit in discrimination between squamous cell carcinoma from mesothelioma.

Metastatic basosquamous carcinoma detected by sentinel lymph node biopsy.

We describe a rare case of basosquamous carcinoma (BSC) in a 69-year-old man. He had noticed a pigmented nodule on his left thigh. Dermoscopic examination showed ulceration at the center and non-typical leaf-like areas at the periphery. Linear-irregular vessels were also seen. He underwent tumor excision in addition to sentinel lymph node biopsy (SLNB). Histopathological examination revealed both features of basal cell carcinoma (BCC) and squamous cell carcinoma. The sentinel lymph node was positive for tumor cells. Immunostaining for Ber-EP4 was focally positive for invasive tumor cells. Although complete inguinal lymphadenectomy had been performed, multiple in-transit metastases around the primary lesion and left iliac lymph node metastasis developed after the operation. It is considered that BSC has a worse prognosis than BCC with a potential for metastasis. Based on the findings of previously reported cases and our case, SLNB may be applied for certain high-risk cases including BSC larger than 3 cm in size and careful follow up is recommended for patients with BSC.

Utility of a limited panel of calretinin and Ber-EP4 immunocytochemistry on cytospin preparation of serous effusions: A cost-effective measure in resource-limited settings.

BACKGROUND: Differentiation between reactive, but morphologically atypical, mesothelial cells and adenocarcinoma in effusions can be problematic. Elaborate immunohistochemical panels have been devised. Techniques like DNA analysis, flow/image cytometry, and K-ras mutation analysis are research oriented and difficult to perform in routine, especially in resource-poor centers. We evaluated the efficacy of a limited two-antibody panel comprising calretinin and Ber-EP4 on cytospin and cell block preparations, in 100 effusion samples. MATERIALS AND METHODS: Fifty cases of reactive mesothelial hyperplasia and 50 cases of adenocarcinoma diagnosed by cytomorphology in ascitic/pleural fluid specimens over a 2-year period were assessed. The diagnoses were confirmed by clinical/histopathologic correlation. Cytospin smears were made in all. Cell blocks were prepared, wherever adequate fluid was available. Immunocytochemistry (ICC) for calretinin and Ber-EP4 was performed. RESULTS: Forty-five of the reactive effusion cases (90%) were calretinin reactive and Ber-EP4 negative. Among the adenocarcinoma cases, 49 (98%) were calretinin negative but Ber-EP4 positive. Thus, both calretinin and Ber-EP4 had a high sensitivity (90% and 98%, respectively), as well as a high specificity (100% and 86%, respectively). In the 21 reactive mesothelial cases, whose cell blocks were made, results were comparable to those on cytospin. However, of the 19 adenocarcinoma cases in which cell blocks were prepared, all were Ber-EP4 immunopositive except for three, which were positive on cytospin, implying false-negative results on cell blocks. CONCLUSIONS: A limited panel of two monoclonal antibodies, calretinin and Ber-EP4, may be useful in cytology, as a "primary antibody panel", for accurate diagnosis and patient management. Additionally, ICC can be performed easily on cytospin preparations, which gave results comparable to cell blocks in our study.

Immunohistochemical Stainning for Ber EP4and EMA in the Diagnosis Basal Cell Carcinoma and Squamous Cell Carcinoma of the Skin / 中华皮肤科杂志

Objective To investigate the significance of immunostainning for Ber EP4and EMA in the diagnosis of basal cell carcinoma and squamous cell carcinoma of the skin.Methods Immunohisto-chemical stainning for Ber EP4and EMA was performed on115cases of basal cell carcinoma,squamous cell carcinoma,Bowen' s disease,actinic keratosis,basosquamous cell carcinoma,seborrheic keratosis,and verruca vulgaris.Specimens were taken from neoplastic tissues as well as the surrounding skin and ap-pendages.Results Ber EP4was positively stained in all cases of basal cell carcinoma and basosquamous cell carcinoma,but negatively stained in squamous cell carcinoma,Bowen's disease,actinic keratosis,sebor-rheic keratosis and verruca vulgaris.Expression of EMA was found in most cases of squamous cell carcinoma and Bowen' s disease,and a few cases of actinic keratosis,and in none of basal cell carcinoma,basosqua-mous cell carcinoma,seborrheic keratosis and verruca vulgaris.Conclusions Routine immunohistochemical staining with both Ber EP4and EMA is helpful for distinction of skin basal cell carcinoma,squamous cell carcinoma,precancerosis and benign hyperplastic dermatoses.