A systematic review of the literature found fifteen articles on the effect of a botulinum toxin on neoplastic cell lines and eight articles on in vivo neoplasms. The reported in vitro effects rely on high doses or the mechanical disruption of cell membranes to introduce the botulinum neurotoxin into the cell cytoplasm. The potency of the botulinum neurotoxin to intoxicate non-neuronal cells (even cell lines expressing an appropriate protein receptor) is several orders of magnitude lower compared to that to intoxicate the primary neurons. The data suggest that the botulinum toxin disrupts the progression of cancer cells, with some studies reporting apoptotic effects. A majority of the data in the in vivo studies also showed similar results. No safety issues were disclosed in the in vivo studies. Limited studies have suggested similar anti-neoplastic potential for the clostridium difficile. New modes of delivery have been tested to enhance the in vivo delivery of the botulinum toxin to neoplastic cells. Careful controlled studies are necessary to demonstrate the efficacy and safety of this mode of anti-neoplastic treatment in humans.

CONTEXT.—:Low-grade fibromatosis-like metaplastic carcinoma (FLMC) is a very rare subtype of triple-negative metaplastic (spindle cell) breast carcinoma. It is characterized by the proliferation of spindle cells closely resembling fibromatosis, which represents a benign fibroblastic/myofibroblastic breast proliferation. Unlike most triple-negative and basal-like breast cancers, FLMC has a very low potential for metastases, but demonstrates frequent local recurrences.OBJECTIVE.—:To genetically characterize FLMC.DESIGN.—:To this end, we analyzed 7 cases by targeted next-generation sequencing for 315 cancer-related genes and performed comparative microarray copy number analysis in 5 of these cases.RESULTS.—:All cases shared TERT alterations (6 patients with recurrent c.-124C>T TERT promoter mutation and 1 patient with copy number gain encompassing the TERT locus), had oncogenic PIK3CA/PIK3R1 mutations (activation of the PI3K/AKT/mTOR pathway), and lacked mutations in TP53. TERT was overexpressed in all FLMCs. CDKN2A/B loss or mutation was observed in 4 of 7 cases (57%). Furthermore, tumors displayed chromosomal stability, with only few copy number variations and a low tumor mutational burden.CONCLUSIONS—:We conclude that FLMCs typically show the recurrent TERT promoter mutation c.-124C>T, activation of the PI3K/AKT/mTOR pathway, low genomic instability, and wild-type TP53. In conjunction with previous data of metaplastic (spindle cell) carcinoma with and without fibromatosis-like morphology, FLMC is most likely distinguished by TERT promoter mutation. Thus, our data support the notion of a distinct subgroup within low-grade metaplastic breast cancer with spindle cell morphology and associated TERT mutations.

Apoplectic leiomyomas-benign uterine leiomyomas with morphologic changes including hemorrhage, hypercellularity, mitotic activity, nuclear atypia, and even necrosis-can be difficult to distinguish from uterine leiomyosarcomas. Apoplectic leiomyomas have been associated with hormonal therapy; however, the relationship between apoplectic leiomyomas, hormones, and ethnicity has not received much attention in the literature. We evaluated the relationship of hormonal therapy and ethnicity in 869 women with uterine leiomyomas, 136 of which qualified as apoplectic leiomyomas.Apoplectic leiomyomas were observed in 23.3% (49/210) of women exposed to hormonal therapy compared to 13.2% (87/659) of women not exposed to hormonal therapy (p < 0.0001). Women taking ethinyl estradiol/norethindrone (Lo-Estrin), leuprolide, and medroxyprogesterone were significantly more likely to have apoplectic leiomyomas compared to women taking other hormonal therapies. Apoplectic leiomyomas were observed in 28.9% (44/152) of African-American women compared to 12.4% (79/639) of Caucasian women (p < 0.0001), and this difference remained statistically significant regardless of hormone use. Apoplectic leiomyomas were observed in 22.1% (77/349) of women ≤ 45 years of age compared to 11.3% (59/520) of women > 45 years of age (p < 0.0001), and this difference remained statistically significant regardless of hormone use.This is the largest study to date examining apoplectic leiomyomas in women on known hormonal therapy compared to women with uterine leiomyomas, but not on hormonal therapy. Information about hormonal therapy, ethnicity, and age can be helpful in the diagnostic interpretation of apoplectic leiomyoma.

In the USA alone, approximately 61,000 new diagnoses of ductal intraepithelial neoplasia 1c-3 (DIN) are made each year. Around 10–20 % of the patients develop a recurrence, about 50 % of which are invasive. Prior studies have shown that invasive breast carcinomas positive for p16 or p53 have a higher frequency of recurrence and a more aggressive course; however, the co-expression of these markers across the entire spectrum of DIN and its potential correlation with grade of the lesions has not been studied previously. Immunohistochemical staining for p16 and p53 was evaluated on 262 DIN lesions from 211 cases diagnosed between 1991 and 2008. The lesions ranged from DIN1b (atypical intraductal hyperplasia) to DIN3 (DCIS, grade 3) and included 45 cases with associated invasive carcinoma. Frequency of staining for both p16 and p53 increased with increasing grade of DIN. Strong co-expression was found exclusively in higher grade DIN lesions (DIN2 and DIN3) particularly those associated with periductal stromal fibrosis and lymphocytic infiltrate. Strong co-expression was seen in 8 of 12 DIN3 lesions (67 %) associated with invasive carcinoma. In conclusion, co-expression of p16 and p53 increases with advancing grade of DIN and is maximal in high grade DIN lesions associated with invasive carcinoma, indicating a more aggressive phenotype. A distinctive variant of DIN with periductal fibrosis and lymphocytic infiltrate invariably falls into the high-grade category, based on either morphology or marker expression. Co-expression of p16/p53 may be of help in distinguishing between high-grade and low-grade DIN lesions.

Pathologists should be aware of the existence of a rare CK7-negative variant of breast carcinoma in general, and of Paget's disease in particular. Cytokeratin 7-negative Paget's disease and CK7-negative ductal intraepithelial neoplasia (ductal carcinoma in situ) present a major diagnostic challenge for pathologists since there is limited awareness of their existence. When there is classic Paget's morphology on H&E sections, GATA3 positivity should resolve any doubts about the diagnosis in the setting of a CK7-negative neoplastic cell population.

Benign apocrine metaplasia (AM) of the adult breast is a very common, but enigmatic lesion. It has been speculated that AM might be a precursor of malignancy or an indicator of a susceptibility of the breast tissue to develop neoplasia, mainly based on comparing the frequency of AM in breast cancer and non-breast cancer patients [1]. Studies using comparative genomic hybridization have supported this by showing similar molecular alterations in benign and malignant apocrine lesions [2]. Few studies, however, have compared expression of biomarkers involved in tumor progression in AM and progressively more advanced atypical apocrine lesions. The expression of C-KIT, COX2, CD24, and CD44s was evaluated by immunohistochemistry in formalin-fixed, paraffin-embedded material of 9 AM, 20 apocrine ductal intraepithelial neoplasia (DIN1c-3) and 40 atypical apocrine lesions (not qualifying for DIN1c-3) and compared to expression of the same biomarkers in adjacent normal ductal epithelium. Of the 66 apocrine lesions, 62 (94 %) did not express C-KIT compared to 4/63 (6 %) of the normal glands (Fisher’s exact, p < 0.001). COX2 was expressed in a significantly higher proportion of apocrine lesions than of normal glands (49 vs. 14 %, p < 0.001), and the number of apocrine lesions positive for CD24 was found to be higher with increasing aggressiveness of the lesions (Spearman, p < 0.001). In conclusion, benign and non-invasive proliferative apocrine lesions of the breast display immuno-phenotypical characteristics previously ascribed mainly to malignant transformation. This could lend support to the theory that AM is an early step towards malignant transformation, albeit associated with slow progression to carcinoma.

Abstract INTRODUCTION: Pubmed database shows no study on pathological features of breast carcinoma in extremely old women (≥90 years). The data post 1980s is limited due to the definition of such age limit as "older than 75". This study redefines age extreme in breast pathology to better correlate with current patient demographics, and provides characteristics of breast cancer in this population. Little information exists about the breast cancer in extremely old women. This investigation elucidates the growing number of breast cancer diagnoses in extremely old women (≥90 years) in a single institution in 2000s. MATERIALS AND METHODS: The database of Yale University Department of Pathology was searched for the terms: "breast carcinoma," "age ≥90" for the past 15 years. Clinicopathologic features of the cases that fit the criteria were studied. RESULTS: A total of 135 patients (134 female; 1 male) aged ≥90 years were identified with a diagnosis of infiltrating carcinoma. A surge in breast cancer diagnoses among elderly patients was noted in 2000s compared to the earlier decade. Only 10 cases were diagnosed between 1990 and 1999 (one case/year) compared to 125 cases diagnosed between 2000 and 2015 (8.3 cases/year). Of these 135 patients, 117 had infiltrating ductal carcinoma (IDC); 16 had infiltrating lobular carcinoma (ILC),1 had pure squamous cell carcinoma. One patient had IDC and ILC ipsilaterally, while another had bilateral IDC. The median age was 92 (range: 90-107), median tumor size was 2.0 cm (range: 0.2-13.0), and the median modified Bloom & Richardson score was 6 (range: 3-9). Among the 117 IDCs, 11 had mucinous, 6 had apocrine, 1 had medullary, 1 had cribriform differentiation. Ductal intraepithelial neoplasia (DCIS) was present in 46% of the cases, while lobular intraepithelial neoplasia (LCIS) was identified in 8% of the cases. CONCLUSIONS: Our data shows an increasing number of breast carcinomas diagnosed among patients ≥90 years of age with a morphological distribution similar to other age groups. This increased frequency is particularly notable in the last 15 years compared to 1990s. While increased life expectancy is a factor, better delivery of screening to elderly patients and patient awareness are important additional contributors to this increase. Extremely old women particularly those in good health may potentially benefit from breast cancer screening. As the number of substantially older patients with breast cancer increases, we have to be prepared to manage the disease in this population. Further studies are warranted to elucidate the frequency of breast cancer in extremely old women and the optimal management that would probably have to be individualized to the patients' health status. Citation Format: Ozerdem U, Tavassoli FA. An impending avalanche-breast cancer among women ≥ 90 years of age. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P1-01-08.

Smooth muscle tumors of the uterus are a diagnostically challenging group of tumors. Molecular surrogate markers reliably distinguishing between benign and malignant tumors are not available. Therefore, the diagnosis is based on morphologic criteria. The aim was to investigate a well-characterized group of challenging uterine smooth muscle tumors consisting of 20 leiomyomas, 13 leiomyomas with bizarre nuclei, and 14 leiomyosarcomas for copy number alterations, MED12 mutations and FH deletions to search for potential diagnostically useful surrogate markers. MED12 mutations were detected in 47, 15, and 25% of leiomyomas, leiomyomas with bizarre nuclei and leiomyosarcomas, respectively. MED12 mutations in leiomyomas with bizarre nuclei were detected outside the hotspot region. FH-deletions were seen in 27, 30.8, and 25% of leiomyomas, leiomyomas with bizarre nuclei and leiomyosarcomas, respectively. By using copy number alteration profiling a clear separation of leiomyomas, leiomyomas with bizarre nuclei and leiomyosarcomas could not be observed. Copy number alterations revealed clear genetic similarities between leiomyomas with bizarre nuclei and leiomyosarcomas. Leiomyosarcomas showed a similar pattern of gains and losses as leiomyomas with bizarre nuclei, with additional copy number alterations and more homozygous losses and high-level amplifications compared to leiomyomas with bizarre nuclei. In conclusion, this study demonstrates that known FH-deletions, a recurrent molecular change in leiomyomas, occur in morphologically challenging variants of leiomyomas, leiomyomas with bizarre nuclei and leiomyosarcomas. Although MED12 mutations are common in leiomyomas, they infrequently occur in leiomyomas with bizarre nuclei and leiomyosarcomas. The genetic similarities between leiomyomas with bizarre nuclei and leiomyosarcomas raise the intriguing possibility that uterine leiomyomas with bizarre nuclei and leiomyosarcomas are closely related and challenge the traditional concept that leiomyoma with bizarre nuclei is a tumor with just marked 'degenerative' cellular changes. These findings support the hypothesis that tumor progression within uterine smooth muscle tumors might occur.

Usual ductal hyperplasia (UDH) of the breast is generally regarded as a nonneoplastic proliferation, albeit loss of heterozygosity has long been reported in a part of these lesions. To gain deeper insights into the molecular drivers of these lesions, an extended mutation profiling was performed. The coding regions of 409 cancer-related genes were investigated by next-generation sequencing in 16 cases of UDH, nine unassociated with neoplasia (classic) and seven arising within papillomas. Phosphatidylinositol 3-kinase/AKT/mammalian target of rapamycin (mTOR) activation was investigated by phosphorylated AKT, mTOR, and S6 immunohistochemistry. Of 16 lesions, 10 (63%) were mutated; 56% of classic lesions were unassociated with neoplasia, and 71% of lesions arose in papillomas. Fourteen missense mutations were detected: PIK3CA [6 (43%) of 14], AKT1 [2 (14%) of 14], as well as GNAS, MTOR, PIK3R1, LPHN3, LRP1B, and IGF2R [each 1 (7%) of 14]. Phosphorylated mTOR was seen in 83% and phosphorylated S6 in 86% of evaluable lesions (phospho-AKT staining was technically uninterpretable). In conclusion, UDH displays mutations of the phosphatidylinositol 3-kinase/AKT/mTOR axis at different levels, with PIK3R1, MTOR, and GNAS mutations not previously described. Specifically, oncogenic G-protein activation represents a yet unrecognized route to proliferation in UDH. On the basis of evidence of activating mutations, loss of heterozygosity, and a mass forming proliferation, we propose that UDH is most appropriately viewed as an early neoplastic intraductal proliferation.

Abstract INTRODUCTION: Ki67 labeling index has been proposed as an independent predictive and prognostic factor in patients with ductal intraepithelial neoplasia (DIN). Ki-67 labeling index of 14% has been suggested as a useful cut-off for stratifying DIN patients for adjuvant radiotherapy and hormonal therapy. No data is available regarding either the distribution pattern of Ki67 immunoreactivity within the ducts involved by DIN or potential correlation of these patterns with lesion grade. In this study, the pattern of distribution of the nuclei immunoreactive with Ki67 was examined in DIN1C (DCIS, grade 1), DIN2 (DCIS, grade2), and DIN3 (DCIS, grade 3) to determine if distinctive patterns could be identified and if these patterns would correlate with lesion grade. METHODS: 47 consecutive DIN cases were retrieved from our departmental files. Of these, 5 qualified as DIN1C, 28 as DIN2 and 14 as DIN3. H&E and Ki67 immunostains were evaluated on each case to elucidate the distribution of Ki67 positive proliferating epithelial cells within the ducts. The DIN cases were evaluated and the patterns of distribution recorded for each case as either basal or haphazard within the epithelial proliferation. Statistical analysis was performed using Chi-square test with Graphpad PRISM statistical analysis software. RESULTS: There was a statistically significant difference between the 3 groups in terms of basal vs haphazard Ki67 immunostaining (Chi –square test, P=0.0001). Basal staining pattern was dominant (100%) among the DIN1c cases, while haphazard staining pattern was the dominant (100%) distribution among the DIN3 cases. One half of the DIN2 cases showed basal staining pattern, while the other half showed a haphazard staining pattern. This feature could be useful in separating DIN lesions into low grade and high grade eliminating grade 2. We also quantified necrosis on a scale of 0 to 2; 0 indicating absence of necrosis and 2 reflecting comedo type necrosis. Necrosis was more common in the ducts with haphazard Ki67 distribution. The extent of necrosis varied significantly between DIN1c, DIN2 and DIN3 (Chi –square test, P<0.0001) CONCLUSIONS: Two distinct patterns of Ki67 immunoreactivity are seen in DIN lesions; the basal pattern is characteristic of DIN1 (low grade DIN), whereas a haphazard pattern is dominant in DIN3 (high grade DIN). These patterns could be used to divide grade 2 DIN into low grade and high grade. This approach is easier and potentially more reproducible than counting the percentage of Ki67 positive cells. The information could be useful from a prognostic standpoint and may well be predictive of potential response to radiation, hormonal and targeted therapies. Citation Format: Ozerdem U, Tavassoli FA. Distribution pattern of Ki67 immunoreactivity in ductal intraepithelial neoplasia (DIN): Correlation with lesion grade and potential utility. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P1-01-07.

Tumor-infiltrating lymphocyte (TIL) count in breast cancer carries prognostic information and represents a potential predictive marker for emerging immunotherapies. However, the distribution of the lymphocyte subpopulations is not well defined. The goals of this study were to examine intratumor heterogeneity in TIL subpopulation counts in different fields of view (FOV) within each section, in different sections from the same biopsy, and between biopsies from different regions of the same cancer using quantitative immunofluorescence (QIF).

Background: In this study, the pattern of distribution of the nuclei immunoreactive with Ki67 was examined in DIN1c (DCIS, grade 1/low grade), DIN2 (DCIS, grade 2/intermediate grade), and DIN3 (DCIS, grade 3/high grade). The lesions were evaluated to determine if distinctive patterns could be identified in correlation with lesion grade.Methods: Fifty seven (n = 57) consecutive DIN cases were investigated. Of these, 15 qualified as DIN1c, 28 as DIN2 and 14 as DIN3. The patterns of distribution were recorded for each case as either basal/peripheral or haphazard within the epithelial proliferation.Results: There was a statistically significant difference between the DIN1c, DIN2 and DIN3 in terms of basal/peripheral versus haphazard distribution of Ki67 immunostaining (Chi-square test, P < 0.0001). Basal/peripheral staining pattern was dominant among the DIN1c cases, while haphazard staining pattern was the dominant distribution among the DIN3 cases. One half of the DIN2 cases showed basal/peripheral staining pattern, while the other half showed a haphazard staining pattern.Conclusion: High grade DIN lesions show haphazard Ki67 staining while low grade DIN lesions show basal/peripheral Ki67 staining in the proliferating epithelial cells. This feature could be practical in separating DIN lesions into low grade (basal/peripheral-Ki67) and high grade (haphazard-Ki67) eliminating the grade 2/intermediate category. (C) 2016 Elsevier GmbH. All rights reserved.

The Breast JournalVolume 22, Issue 4 p. 463-464 Breast Images An Interesting Presentation of Ischemic Septal Panniculitis of the Breast Natalie R. Simmons MD, Corresponding Author Natalie R. Simmons MD Department of Radiology, Yale University, New Haven, ConnecticutAddress correspondence and reprint requests to: Natalie R. Simmons, Department of Radiology, Yale University, 333 Cedar Street, New Haven CT 06510, USA, or e-mails: natalie.simmons@aya.yale.edu and nsimmons@stanfordalumni.orgSearch for more papers by this authorUgur Ozerdem MD, Ugur Ozerdem MD Department of Radiology, Yale University, New Haven, ConnecticutSearch for more papers by this authorLiane E. Philpotts MD, Liane E. Philpotts MD Department of Radiology, Yale University, New Haven, ConnecticutSearch for more papers by this authorFattaneh Tavassoli MD, Fattaneh Tavassoli MD Department of Radiology, Yale University, New Haven, ConnecticutSearch for more papers by this author Natalie R. Simmons MD, Corresponding Author Natalie R. Simmons MD Department of Radiology, Yale University, New Haven, ConnecticutAddress correspondence and reprint requests to: Natalie R. Simmons, Department of Radiology, Yale University, 333 Cedar Street, New Haven CT 06510, USA, or e-mails: natalie.simmons@aya.yale.edu and nsimmons@stanfordalumni.orgSearch for more papers by this authorUgur Ozerdem MD, Ugur Ozerdem MD Department of Radiology, Yale University, New Haven, ConnecticutSearch for more papers by this authorLiane E. Philpotts MD, Liane E. Philpotts MD Department of Radiology, Yale University, New Haven, ConnecticutSearch for more papers by this authorFattaneh Tavassoli MD, Fattaneh Tavassoli MD Department of Radiology, Yale University, New Haven, ConnecticutSearch for more papers by this author First published: 18 April 2016 https://doi.org/10.1111/tbj.12607Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat No abstract is available for this article. Volume22, Issue4July/August 2016Pages 463-464 RelatedInformation