Invasive Malignant Lesions Research Articles

Prevalence, risk factors and genotype distribution of human papillomavirus infection among women with and without invasive cervical cancer: Findings from a hospital-based study in Bihar, India.

Background Human papillomavirus (HPV) infection is largely responsible for the development of invasive cervical cancer (ICC). Its prevalence, risk factors and genotype distribution among women residing in Bihar (third most populous Indian state) with and without ICC are not well known. Methods In this hospital-based study, we followed up 1439 participants with cytology and HPV report. HPV detection and genotyping were performed using the TaqMan-based real-time PCR method. Clinical and sociodemographic data were collected and analysed using statistical methods. Results The overall prevalence of HPV infection was 37.3% (537/1439) and 11 different types of HPV genotypes were observed. Higher HPV positivity was found in premalignant, intraepithelial and invasive malignant lesions of the cervix; 73.8% (93/126) of atypical squamous cells, cannot exclude high-grade squamous intraepithelial lesion (ASC-H) and high-grade squamous intraepithelial lesions (HSIL) and 93.4% (114/122) of invasive malignancies were infected with HPV in comparison to only 26.1% (245/938) of negative for intraepithelial lesion or malignancy (NILM) cytology. Moreover, HPV was found in 95.2% (236/248) of histologically confirmed cases of carcinoma cervix. HPV16 and HPV18 infections were reported in 78.2% (194/248) and 8.9% (22/248), respectively. The remaining patients had infection with other high-risk strains/co-infection with multiple strains or were HPV-negative. Various socio-demographic factors including women >50 years of age, >10 years of marriage and high parity were significantly associated with HPV infection. Conclusion Our data suggest that HPV16 infection may be the major cause for ICC among women residing in Bihar. Our findings may serve as a baseline for developing an appropriate screening and vaccination strategy for Bihar.

Oncocytic Follicular Cell-Derived Thyroid Tumors With Papillary Growth Pattern: A Clinicopathologic Study of 32 Cases.

Oncocytic thyroid tumors displaying a papillary growth pattern are rare and may cause diagnostic problems. To examine the clinicopathologic features of a series of 32 follicular cell-derived tumors composed of cells with oncocytic cytoplasm and displaying papillary architecture. Thirty-two cases were collected and studied to assess clinicopathologic features, including immunohistochemical and molecular testing for BRAF V600E. The patients were 26 women and 6 men, aged 17 to 77 years. The nodules ranged from 0.3 to 6.0 cm. Eighteen cases showed features of oncocytic hyperplastic nodules and were identified against a background of thyroid follicular nodular disease; 4 cases showed features of oncocytic follicular adenoma; and 10 cases corresponded to carcinomas with oncocytic and papillary features. Nuclear features of papillary thyroid carcinoma were absent or exceedingly rare. All cases were negative for HBME-1 and cytokeratin 19 (CK19) and wild type for BRAF V600E. Follow-up in 25 patients showed that all patients with hyperplastic nodules and oncocytic follicular adenomas were alive and well and free of disease from 7 to 20 years. One patient with oncocytic follicular carcinoma showed metastases and died of tumor at 16 months; 2 patients with carcinoma had metastases and recurrence at 6 and 7 years; and 5 patients with invasive tumors were free of disease from 5 to 10 years. Oncocytic thyroid tumors with papillary features can span a spectrum from benign hyperplastic, to encapsulated neoplastic, to invasive malignant lesions. Owing to their papillary features, it is important not to confuse them for other types of thyroid tumors, such as oncocytic papillary thyroid carcinoma.

Noninvasive diagnosis of pulmonary nodules using a circulating tsRNA-based nomogram.

Evaluating the accuracy of pulmonary nodule diagnosis avoids repeated low-dose computed tomography (LDCT)/CT scans or invasive examination, yet remains a main clinical challenge. Screening for new diagnostic tools is urgent. Herein, we established a nomogram based on the diagnostic signature of five circulating tsRNAs and CT information to predict malignant pulmonary nodules. In total, 249 blood samples of patients with pulmonary nodules were selected from three different lung cancer centers. Five tsRNAs were identified in the discovery and training cohorts and the diagnostic signature was established by the randomForest algorithm (tRF-Ser-TGA-003, tRF-Val-CAC-005, tRF-Ala-AGC-060, tRF-Val-CAC-024, and tiRNA-Gln-TTG-001). A nomogram was developed by combining tsRNA signature and CT information. The high level of accuracy was identified in an internal validation cohort (n = 83, area under the receiver operating characteristic curve [AUC] = 0.930, sensitivity 100.0%, specificity 73.8%) and external validation cohort (n = 66, AUC = 0.943, sensitivity 100.0%, specificity 86.8%). Furthermore, the diagnostic ability of our model discriminating invasive malignant ones from noninvasive lesions was assessed. A robust performance was achieved in the diagnosis of invasive malignant lesions in both training and validation cohorts (discovery cohort: AUC = 0.850, sensitivity 86.0%, specificity 81.4%; internal validation cohort: AUC = 0.784, sensitivity 78.8%, specificity 78.1%; and external validation cohort: AUC = 0.837, sensitivity 85.7%, specificity 84.0%). This novel circulating tsRNA-based diagnostic model has potential significance in predicting malignant pulmonary nodules. Application of the model could improve the accuracy of pulmonary nodule diagnosis and optimize surgical plans.

Abstract LB111: Epigenetic mechanism underlying pathogenesis of skin photo-damage and actinic keratoses

Abstract Chronic skin exposure to solar ultraviolet B (UVB) radiation causes cutaneous inflammation, sun-burns and pre-malignant lesions known as Actinic Keratosis (AK). About 20% of AKs may progress to highly invasive malignant lesions known as Squamous Cell Carcinoma (SCC). These lesions carry multiple mutations in oncogenes and/or tumor suppressor genes. However, the molecular mechanism underlying AK is not incompletely defined. Here, we demonstrate that cutaneous UVB exposure leads to activation of bromodomain-4 (BRD4), an epigenetic modifier. We observed enhanced expression of BRD4 and its hyper-acetylation marks, H3K18ac and H3K27ac in AK, accompanied by elevated transcription of BRD4-target genes: MMP13, CXCL9, IL33, Apol19b, IL19 and Oasl2. Using confocal microscopy, we observed enhanced nuclear accumulation of BRD4 and H3K27ac in the sun-exposed lesional skin, AK and in SCC tissues from patients. Ptch+/−/SKH1 mice irradiated with to chronic UVB manifest similar molecular alterations in highly photo-damaged skin. By manipulating BRD4 expression in human skin SCC A431 cells, we identified two important BRD4-regulated signaling pathways, which may be involved in the molecular pathogenesis of AKs and SCCs. BRD4-mediated inflammatory cytokine signaling is regulated by EZH2-SOCS3-NFkB/STAT3 axis while Myc-Survivin-CDK4-E2F1 axis drives tumor cell hyper-proliferation. Co-localization of BRD4/H3K27ac and Survivin-E2F1 was contiguous from sun-burn skin to AK and SCC. We observed enhanced copy number of Myc in human AK lesional skin and associated SCCs. We also show that Myc-Survivin-CDK4-E2F1 axis is critical in driving the progression of tumor keratinocytes to highly invasive and fast growing SCCs. This signaling acts by enhancing the expression epithelial-mesenchymal transition transcription factors and other proteins involved in UVB-induced SCC development. 9cUAB30, a RXR agonist developed by us blocked BRD4 activation and its downstream signaling in A431 and SCC13 cells. Oral administration of 9cUAB30 abrogates UVB-induced inflammation and keratinocyte hyper-proliferation in murine skin in BRD4 dependent manner. 9cUAB30 treatment attenuates growth of human SCC xenograft tumors in highly immune-suppressed mice, a response which was significantly more profound when the drug was administered with a survivin inhibitor, LQZ-7I. These translational studies demonstrate that UVB-induced cutaneous BRD4 drives AK pathogenesis and their progression to invasive SCCs. Citation Format: Mahendra P. Kashyap, Rajesh Sinha, Rubina Shafi, Venkatram R. Atigadda, Craig A. Elmets, Mohammad Athar. Epigenetic mechanism underlying pathogenesis of skin photo-damage and actinic keratoses [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 2 (Clinical Trials and Late-Breaking Research); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(8_Suppl):Abstract nr LB111.

Dental MRI of Oral Soft-Tissue Tumors-Optimized Use of Black Bone MRI Sequences and a 15-Channel Mandibular Coil.

Soft-tissue lesions in the oral cavity, one of the most common sites for tumors and tumor-like lesions, can be challenging to diagnose and treat due to the wide spectrum from benign indolent to invasive malignant lesions. We report an abnormally large, rapidly growing hyperplastic lesion originating from the buccal mucosa in a 28-year-old male patient. Clinical examination revealed a well-circumscribed, smooth-surfaced, pinkish nodular lesion measuring 2.3 × 2 cm, which suggested the differential diagnosis of irritation fibroma, pyogenic granuloma, oral lipoma, and other benign or malignant neoplasms such as hemangioma, non-Hodgkin’s lymphoma, or metastases to the oral cavity. Dental MRI using a 15-channel mandibular coil was performed to improve perioperative radiological and surgical management, avoiding adverse intraoperative events and misdiagnosis of vascular malformations, especially hemangiomas. Black bone MRI protocols such as STIR (short-tau inversion recovery) and DESS (double-echo steady-state) were used for high-resolution radiation-free imaging. Radiologic findings supported the suspected diagnosis of an irritation fibroma and ruled out any further head and neck lesions, therefore complete surgical resection was performed. Histology confirmed the tentative diagnosis. This article evaluates the use of this novel technique for MR diagnosis in the perioperative management of soft-tissue tumors in oral and maxillofacial surgery.

A90 WHEN DO TRAINEES ACHIEVE COMPETENCY IN PERFORMING ENDOSCOPIC SUBMUCOSAL DISSECTION: A SYSTEMATIC REVIEW

BackgroundEndoscopic submucosal dissection (ESD) is an established organ sparing curative endoscopic resection technique for the management of pre-malignant and superficially invasive malignant lesions of the gastrointestinal tract. However, little is understood of its learning curve with suggested competency measures including en bloc resection, R0 resection, adverse events and resection speed.AimsWe aimed to perform a systematic review on when competency is achieved in ESD.MethodsTwo authors independently searched MEDLINE and EMBASE (1946 to Aug 2021) for full-text original citations including grey literature assessing the ESD learning curve. A learning curve was defined as an assessment of competency as a function of increasing trainee experience. Thresholds for competency were defined as en bloc resection ≥ 80%, R0 resection ≥ 80%, perforation rate ≤ 5% and resection speed ≤ 6.67min/cm2.ResultsForty-three studies (1 esophageal, 11 gastric, 27 colorectum, 4 multiple sites) with 157 trainees and 8780 ESD procedures were included. Baseline experience in esophagogastroduodenoscopy, colonoscopy, endoscopic mucosal resection and ESD were 800–10,000, 100–10,000, 4–700 and 0–300 procedures, respectively. 16 studies used animal model training prior to assessing the ESD learning curve.En bloc resection, R0 resection, perforation rate and resection speed were used as markers of competency in 33, 29, 28 and 21 studies, respectively. When pooling evaluations where competency was achieved, it was reached in the esophagus, stomach, colorectum and for multiple sites between <10, <10–150, <10–301, and <20–300 procedures, respectively (Table 1). However, competency in R0 resection, perforation rate, and resection speed was not uniformly achieved within 12 studies.ConclusionsCompetency in ESD can be achieved in <10 - 399 procedures, with variability dependent on organ site and baseline level of training. With the widespread adoption of ESD, standardization of training and the assessment of competency are needed.Table 1. ESD competency outcomes by site and number of proceduresCompetency AchievedEn bloc ≥ 80%R0 ≥ 80 %Perforation Rate ≤ 5%Resection Speed ≤ 6.67min/cm2Esophagus<10<10<10-Stomach<10 - 38<12 - 100<10 - 150-Colorectum<12 - 180<16 - 301<10 - 25025 - 240Multiple Sites<20 - 20021 - 30025 - 36300Funding AgenciesNone

Efficacy of MRI-Directed Breast Ultrasound and Lesion Characteristics Affecting Visualization on Breast Ultrasound

ObjectiveTo assess the efficacy of magnetic resonance imaging (MRI)-directed ultrasound (US) in further characterizing lesions detected on MRI and to evaluate the correlation between lesion characteristics and their visibility on US. MethodsA retrospective review was performed of the records of patients with breast abnormalities detected on MRI between May 2018 and December 2020. All patients who were given a Breast Imaging-Reported and Data System Assessment Category 4 or 5 on breast MRI and sent to breast US for assessment of the MRI abnormality were included. Patients lost to follow-up or those who did not get their subsequent ultrasound at our institution were excluded. The following factors were recorded for all lesions described on the MRI reports: breast density, degree of background parenchymal enhancement, type of MRI finding (mass, areas of non-mass enhancement [NME], enhancing focus), largest dimension, T2 signal intensity, kinetics, shape, margins, enhancement pattern, and presence of ipsilateral breast malignancy. For the subset of the lesions that were identified on MRI-directed US, we recorded the following sonographic features: largest dimension, shape, and margin. The chi-square or Fisher exact tests of association were used to compare categorical characteristics. ResultsTwo hundred and thirty-three MRI-detected breast lesions were included in the study, which consisted of 171 masses, 7 enhancing foci, and 55 NME lesions. Of all lesions, 52% (89/171) of masses, 29% (16/55) of areas of NME, and 29% (2/7) of foci were seen on US. Moreover, 43 per 63 (68%) of malignant, 18 per 47 (38%) of high-risk and 46 per 123 (37%) of benign lesions were seen on MRI-directed US. Of the invasive malignant lesions, 77% (36/47) had a sonographic correlate. For masses, the average size on MRI was 1.1 cm, and MRI size had a statistically significant association with US visibility (P value 0.01). Similarly, spiculated margins on MRI for masses had a significant association with US visibility (P value 0.0006). Linear, segmental, and regional distributions of areas of NME were associated with decreased likelihood of sonographic visibility (P = 0.049). ConclusionMRI-directed US is a useful tool in the management of MRI-detected breast lesions. Our study indicates that masses, particularly those with spiculated margins on MRI, should be sent to US for further assessment. Additionally, masses are more likely to be seen under US as their size increases. MRI-directed US is less likely to visualize areas of NME and enhancing foci, which may be managed with MRI follow-up or MRI-guided biopsy depending on their level of suspicion.

Clinical Application of Computer-Aided Diagnosis for Breast Ultrasonography: Factors That Lead to Discordant Results in Radial and Antiradial Planes.

ObjectiveTo investigate factors that may lead to discordant results in radial and antiradial planes when applying a computer-aided diagnostic system (S-DetectTM) for breast ultrasound (US).MethodsFrom May 2019 to September 2019, a total of 288 breast lesions from 286 consecutive women were analyzed. Diagnostic performance and diagnostic agreement of the CAD system between the radial and antiradial planes were calculated. Based on the CAD results in the radial and antiradial planes, the lesions were classified into two groups: the agreement group and the discordant group. Ultrasound imaging and clinicopathologic factors in the two groups were compared.ResultsOf the 288 breast lesions, 187 (64.7%) were benign, and 101 (35.3%) were malignant. There was no difference in diagnostic performance of the CAD system between the radial and antiradial planes. The diagnostic agreement of the CAD system between these two orthogonal planes was good (ĸ = 0.645). Compared to category 3, 4C and 5 lesions, category 4A and 4B lesions were more likely to have discordant CAD results. In the subgroup of malignant tumors, lesions diagnosed as carcinoma in situ (P = 0.014), staged as T1 (P = 0.013) and with low Ki-67 status (P = 0.024) were significantly associated with discordant CAD results.ConclusionThe CAD system for breast ultrasound has a favorable diagnostic performance, and discordant CAD results between the radial and antiradial planes were more frequent for BI-RADS category 4A-4B lesions and less invasive malignant lesions.

BASIC (BLI Adenoma Serrated International Classification) classification for colorectal polyp characterization with blue light imaging.

Advanced endoscopic imaging has revolutionized the characterization of lesions during colonoscopy. The aim of this study was to create a new classification for differentiating subcentimetric hyperplastic and adenomatous polyps, and deeply invasive malignant lesions using blue-light imaging (BLI) with high definition, with and without optical magnification, as well as to assess its interobserver concordance. A video library consisting of 48 videos/still images (with/without optical magnification) from 24 histologically verified polyps/cancer with BLI was prospectively created. In the first step, seven endoscopists with experience in electronic chromoendoscopy reviewed 12 BLI videos/still images with/without magnification representative of the different histotypes, and individually identified possible descriptors. In the second step, these descriptors were categorized and summarized with a modified Delphi methodology. In the third step, the seven endoscopists independently reviewed the remaining 36 videos/still images with/without optical magnification, and the interobserver agreement for the new descriptors was assessed. The interobserver agreement between endoscopists was assessed using Gwet's AC1. By reviewing the initial 12 videos/still images, 43 descriptors were proposed. By a modified Delphi process, the endoscopists eventually agreed on summarizing 12 descriptors into three main domains. The main domains identified were: polyp surface (mucus, yes/no; regular/irregular; [pseudo]depressed, yes/no), pit appearance (featureless, yes/no; round/nonround with/without dark spots; homogeneous/heterogeneous distribution with/without focal loss), and vessels (present/absent, lacy, pericryptal, irregular). Interobserver agreement for the polyp surface domain appeared to be almost perfect for mucus (AC1 0.92 with and 0.88 without optical magnification), substantial for the regular/irregular surface (AC1 0.67 with and 0.66 without optical magnification). For the pit appearance domain, interobserver agreement was good for featureless (AC1 0.9 with and 0.8 without optical magnification), and round/nonround (AC1 0.77 with and 0.69 without optical magnification) descriptors, but less consistent for the homogeneity of distribution (AC1 with/without optical magnification 0.58). Agreement was almost perfect for the vessel domain (AC1 0.81 - 0.85). The new BASIC classification takes into account both morphological features of the polyp, as well as crypt and vessel characteristics. A high concordance among the observers was shown for most of the summarized descriptors. Optical magnification had a beneficial effect in terms of interobserver agreement for most of the descriptors.

Elastosis in premalignant and malignant lesions of breast-A histopathological and histochemical study

Background: Stromal changes in all the premalignant, insitu/ microinvasive and invasive malignant lesions of breast helps in better understanding of disease process at the level of the tumor and host stromal reaction. The pattern of elastosis even correlates better with prognosis than the predicted behaviour of tumour cells. Aims and Objectives: The present study is undertaken to emphasize the importance of stromal changes (elastosis) and to predict the prognosis and behaviour of tumor cells. Study design: The study was conducted on 90 patients of breast lesions (66 cases of invasive malignant breast lesions and 04 cases of insitu breast lesions and 20 cases of proliferative breast lesions). Material and Methods: The tissue for histopathological studies were collected in 10 % buffered formalin solution .The paraffin wax blocks were made and 4-5 µ thick sections were cut and stained by H & E and special histochemical stains. Result: Moderate degree of elastosis was found to be associated with maximum number of invasive malignant breast lesions Proliferative breast lesions and insitu malignant lesions showed mild to moderate degree of elastosis. Consistent results were obtained in Grade II malignant lesion where moderate to marked degree of elastosis was seen. Conclusion: In all cases of early invasive and invasive malignant lesions of breast, elastosis as a indicator of stromal change should be carefully studied and included in predicting the final outcome along with histological types to get the better results.

Breast lesions of uncertain malignant nature and limited metastatic potential: proposals to improve their recognition and clinical management.

Breast lesions comprise a family of heterogeneous entities with variable patterns of presentation, morphology and clinical behaviour. The majority of breast lesions are classified traditionally into benign and malignant conditions and their behaviour can, in the vast majority of cases, be predicted with a reasonable degree of accuracy. However, there remain lesions which show borderline features and lie in a grey zone between benign and malignant, as their behaviour cannot be predicted reliably. Defined pathological categorization of such lesions is challenging, and for some entities is recognized to be subjective and include a range of diagnoses, and forms of terminology, which may trigger over- or undertreatment. The rarity of these lesions makes the acquisition of clinical evidence problematic and limits the development of a sufficient evidence base to support informed decision-making by clinicians and patients. Emerging molecular evidence is providing a greater understanding of the biology of these lesions, but this may or may not be reflected in their clinical behaviour. Herein we discuss some breast lesions that are associated with uncertainty regarding classification and behaviour, and hence management. These include biologically invasive malignant lesions associated with uncertain metastatic potential, such as low-grade adenosquamous carcinoma, low-grade fibromatosis-like spindle cell carcinoma and encapsulated papillary carcinoma. Other lesions of uncertain malignant nature remain, such as mammary cylindroma, atypical microglandular adenosis, mammary pleomorphic adenoma and infiltrating epitheliosis. The concept of categories of (1) breast lesions of uncertain malignant nature and (2) breast lesions of limited metastatic potential are proposed with details of which histological entities could be included in each category, and their management implications are discussed.

Addition of parity to the risk of malignancy index score in evaluating adnexal masses

ObjectiveThe aim of our study was to evaluate the individual contribution of parity when incorporated as another parameter into the four risk of malignancy indices (RMI 1–4) to differentiate noninvasive benign lesions from invasive malignant ovarian lesions. Materials and methodsAfter calculating RMI 1–4 for each patient included in this study, the resulting RMI scores were further multiplied by the parity score (P) of each patient to calculate the RMI parity (RMIP) score. ResultsA cutoff value of 300 for RMIP 1 yielded 95.0% specificity, 97.4% negative predictive value (NPV), 88.5% sensitivity, and 79.3% positive predictive value (PPV) and performed better than RMI 1 in the preoperative diagnosis of invasive malignant lesions. RMIP 2 with a cutoff value of 400 yielded 95.0% specificity, 97.4% NPV, 88.5% sensitivity, and 79.3% PPV, and it also performed better than RMI 2. A cutoff value of 400 for RMIP 3 provided 97.5% specificity, 97.5% NPV, 88.5% sensitivity, and 88.5% PPV and performed better than RMI 3. However, a cutoff value of 400 for RMIP 4 provided 90.0% specificity, 97.3% NPV, 88.5% sensitivity, and 65.7% PPV but did not perform better than RMI 4 in the preoperative diagnosis of invasive malignant lesions. ConclusionRMIP 1–3 scales were more reliable tools for the preoperative diagnosis of invasive adnexal masses compared with the traditional RMI 1–3 scales.

Should Cut-Off Values of the Risk of Malignancy Index be Changed for Evaluation of Adnexal Masses in Asian and Pacific Populations?

The risk of malignancy index (RMI) for the evaluation of adnexal masses is a sensitive tool in certain populations. The best cut off value for RMI 1, 2 and 3 is 200. The cut off value of RMI-4 to differentiate benign from malignant lesions is 450. Our aim was to evaluate the efficiency of four different malignancy indexes (RMI1-4) in a homogeneous population. We evaluated a total of 153 non-pregnant women with adnexal masses who did not have a history of malignancy and who were above 18 years of age. A cut-off value of 250 for RMI-1 provided 95.9% inter-observer agreement, yielding 95.9% specificity, 93.5% negative predictive value, 75.0% sensitivity and 82.8% positive predictive value. A cut-off value of 250 for RMI-1 showed high performance in preoperative diagnosis of invasive malignant lesions than cut-off value of 200 in our population. A cut-off value of 350 for RMI-2 provided 94.5% inter-observed agreement, yielding 94.2% specificity, 93.4% negative predictive value, 75.0% sensitivity and 77.4% positive predictive value. RMI-2 showed the higher performance when the cut-off value was set at 350 in our population. A cut-off value of 250 provided 95.2% inter-observer agreement, yielding 95.0% specificity, 93.2% negative predictive value, 75.0% sensitivity, and 88.0% positive predictive value. RMI-3 showed the highest performance to diagnose malignant adnexal masses when the cut-off value was set at 250. In our study, RMI-4 showed similar statistical performance when the cut-off value was set at 400 [(Kappa: 0.684/p=0.000), yielding 93.8% inter-observer agreement, 93.4% specificity, 93.4% negative predictive value, 75.0% sensitivity, and 75.0% negative predictive value]. We showed successful utilization of RMIs in preoperative differentiation of benign from malignant masses. Many studies conducted in Asian and Pacific countries have reported different cut-off values as was the case in our study. We think that it is difficult to determine universally accepted cut-off values for RMIs for common use around the globe.

Abstract 2497: Upregulation of annexin A8 is a common molecular phenotype of aberrant breast acinar morphogenesis induced by different genetic factors.

Abstract It is estimated that up to 90% of women have benign breast lesions, some of which are pre-malignant. In situ lesions, particularly the ones that involve the ducts (ductal carcinoma in situ, DCIS), have the potential to progress into invasive malignant lesions and become life threatening. These breast premalignant states are identified because of specific morphological tissue abnormalities. Three-dimensional (3D) cultures of human mammary epithelial cells closely recapitulate breast acinar morphogenesis. While untransformed mammary epithelial cells form hollow acini, lined by polarized, non-proliferating cells, breast cancer cells form acini filled with non-polarized, actively proliferating cells, similar to DCIS. By using a proteomic approach in a panel of morphologically aberrant acini derived from human breast epithelial cells, we found that the disruption of acinar morphogenesis by different genetic factors generates a common proteomic signature. Among the proteins that we found consistently upregulated, we identified Annexin A8 (ANXA8), a member of the annexin family of calcium and membrane-binding proteins with a wide range of cellular functions, including cell division, apoptosis, and cell growth regulation. We tested ANXA8 expression in tissue samples from patients with atypical ductal hyperplasia (ADH), and tissue microarrays of DCIS, with or without, history of invasive breast cancer. Interestingly, this analysis detected ANXA8 increased expression in atypical proliferating ducts relative to non-proliferative ducts, making ANXA8 a potential marker of breast pre-malignancies. Acknowledgments: this study was funded by grants of the Susan Komen Foundation, the Breast Cancer Coalition of Rochester, the Friends for an Earlier Test of Breast Cancer Foundation. Citation Format: Stefano Rossetti, Francesca Corlazzoli, Nicoletta Sacchi. Upregulation of annexin A8 is a common molecular phenotype of aberrant breast acinar morphogenesis induced by different genetic factors. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2497. doi:10.1158/1538-7445.AM2013-2497 Note: This abstract was not presented at the AACR Annual Meeting 2013 because the presenter was unable to attend.

Cytoplasmic and stromal expression of laminin γ 2 chain correlates with infiltrative invasion in ovarian mucinous neoplasms of gastro-intestinal type

Ovarian mucinous neoplasms of gastro-intestinal type (GI-type) are known to be a heterogeneous tumor composed of benign, borderline and non-invasive and invasive malignant lesions. The presence of infiltrative invasion is also known to be an important prognostic factor of this neoplasm. Laminin γ 2 chain, known to stimulate tumor cell invasion and migration, has not been sufficiently investigated in ovarian mucinous neoplasms. The purpose of this study was thus to clarify the role of laminin γ 2 in ovarian mucinous neoplasms of GI-type. We selected each morphological phase of tumor development from 61 cases of mucinous neoplasms of the GI-type: 55 adenoma lesions, 60 borderline lesions, 20 microinvasive lesions, 17 intraepithelial carcinoma lesions, 38 expansile invasive carcinoma lesions, 19 infiltrative invasive carcinoma lesions and 5 mural nodules lesions; and evaluated the localization of laminin γ 2 in the lesions using immunohistochemical method. The staining pattern was classified into i) basement membranous (BM), ii) cytoplasmic (CYT) and iii) stromal (S) pattern. The BM pattern was characteristic in adenoma, borderline, and interaepithelial and expansile invasive carcinoma lesions. The CYT and S patterns were characteristic in infiltrative invasive lesions. The staining pattern of mural nodules was similar to that of infiltrative invasion. The infiltrative invasion of GI-type ovarian mucinous neoplasms may be promoted by cytoplasmic and/or stromal expression of laminin γ 2 chain.

Use of ultrasound pattern recognition by expert operators to identify borderline ovarian tumors: a study of diagnostic performance and interobserver agreement

To assess the accuracy and reproducibility of ultrasound 'pattern recognition' for the diagnosis of borderline ovarian tumors by asking experienced ultrasound operators to evaluate representative images of different types of adnexal tumor. Digitally stored static two-dimensional B-mode images of representative cases of benign, borderline and invasive malignant ovarian tumors were independently assessed by three expert sonologists who had not performed the original real-time ultrasound examination. The outcome measures included diagnostic accuracy and interobserver agreement in the diagnosis of benign, borderline or invasive malignant ovarian tumors. One hundred and sixty-six cases were included in the final data analysis. A correct classification was made by all three experts in 83% of the primary invasive cancers, 76% of the benign masses and in 44% of the borderline malignant tumors (P < 0.01). The experts showed a tendency to misclassify borderline tumors as benign rather than primary invasive (ratio of 8 : 1 for Expert A, 4 : 1 for B and 6 : 1 for C). The interobserver agreement between any two experts was very good when they were tested for their ability to discriminate between invasive and non-invasive (benign and borderline) ovarian tumors (Cohen's kappa 0.85-0.88), but poorer for the discrimination between malignant (invasive and borderline) and benign tumors (kappa 0.70-0.78). The accuracy of ultrasound diagnosis of borderline tumors is lower in comparison with benign and invasive malignant lesions. The diagnostic performance and interobserver agreement are better when the outcomes are dichotomized into non-invasive and invasive malignant lesions, as opposed to the traditional diagnosis of benign and malignant tumors.

Investigation of human papillomavirus by hybrid capture II in cervical carcinomas including 113 adenocarcinomas and related lesions

Hybrid capture is an easy and highly sensitive technique for screening population due to its capacity to detect malignant and premalignant lesions of the cervix. To evaluate its sensitivity, we investigated the frequency of high-risk human papillomavirus (HPV) infection and its correlation with glandular malignant lesions, analyzing a total of 113 cases of adenocarcinomas and related lesions. High-risk HPV was investigated using a hybrid capture II (HC2) assay. Samples were collected in two different ways: either brushed directly from surgical specimens before fixation or collected from the patients. We also investigated the frequency of HPV in squamous malignant lesions, 65 squamous cell carcinomas (SCC) and 66 in situ squamous cell carcinomas (ISSCC), to compare the occurrence of HPV for these lesions. The 113 glandular lesions comprised 62 invasive adenocarcinomas (IAC), 8 in situ adenocarcinomas (ISAC), 26 IAC plus SCC, and 17 adenosquamous cells carcinomas (ASCC). The HPV-positive reactions were as follows: 51 (82.2%) in IAC, 8 (100%) in ISAC, 25 (96.1%) in IAC plus SCC, and 14 (82.3%) in ASCC. HC2-positive results in the squamous malignant lesions were as follows: 58 of 63 (89.0%) for SCC and 94 of 103 (91.2%) for ISSCC. High-risk HPV infection was quite similar for glandular and pure squamous invasive malignant lesions, 82.2% and 89.0%, respectively, indicating that high-risk HPV is also highly prevalent in glandular lesions. Although hybrid capture proved to be an excellent adjunctive technique, we do not believe its results merit replacing the Pap smear as a screening tool.

The value of detection of normal ovarian tissue (the ‘ovarian crescent sign’) in the differential diagnosis of adnexal masses

The aim of the study was to evaluate whether the presence of normal ovarian tissue adjacent to an adnexal tumor (the 'ovarian crescent sign') could assist in the preoperative differential diagnosis of adnexal lesions. This was a prospective observational study including 100 women with a preoperative diagnosis of an adnexal mass. Demographic and biochemical data were collected and all women underwent a detailed transvaginal ultrasound scan. Tumor volume, morphological characteristics and Doppler features were recorded in each case. In addition, the tissue adjacent to the cyst was systematically examined for the presence of normal ovarian tissue. All the findings were compared to the final histological diagnosis. Sixty-seven (67%) of the cysts removed were benign, nine (9%) were borderline and 24 (24%) women had invasive malignant lesions. Normal ovarian tissue was seen in 58/76 (76%) women with non-invasive lesions, and in one woman (4%) with an invasive malignancy. In the absence of normal ovarian tissue, ovarian cancer was diagnosed with a sensitivity of 96% and specificity of 76%. The presence of normal ovarian tissue adjacent to an ovarian cyst is a useful morphological feature that may be used to help exclude an invasive ovarian malignancy in women with adnexal masses detected on ultrasound scan.

The utility of toluidine blue application as a diagnostic aid in patients previously treated for upper oropharyngeal carcinoma

Objectives. This study evaluated the utility (usefulness) of toluidine blue application as an aid to the recognition and diagnosis of clinically evident lesions in a series of patients previously treated for oral cancer and monitored in a cancer center. In addition to increased risk of recurrence of cancer or new second primary lesions, patients who have had previous treatment for oropharyngeal cancer may be more difficult to assess because of tissue changes that occur as a result of previous radiation therapy. Study design. Patients with a history of oral malignancy were assessed by clinical examination followed by application of toluidine blue. Biopsy sites were determined on the basis of unaided visual examination and by the findings on toluidine blue application. Biopsy specimens were reviewed by a pathologist blinded to the clinical findings. Results. Unaided clinical examination identified 78% of carcinoma in situ or invasive malignant lesions compared with toluidine blue application, which identified all (100%) carcinoma in situ or invasive malignant lesions ( p = 0.02) and produced no false-negative findings. No differences were found between clinical examination and toluidine application in the detection of dysplastic lesions. Conclusion. Toluidine blue retention was seen in all cases of carcinoma in situ and invasive carcinoma, and no false-negative findings were seen with toluidine blue. When used by a trained and experienced clinician in a cancer center, toluidine blue was a valuable visual aid to clinical examination of oral mucosal lesions.

Carcinogen-induced changes in tracheal epithelium cultured in serum-free, chemically defined medium.

Rat tracheal tissue was cultured for periods up to 2 months in medium containing benzo[a]pyrene, and the epithelium was studied for the histologic effects of deletion of serum from the medium. Nodular hyperplasia occurred in 3 days in the absence of serum, but was not seen until several weeks of culture in media containing the same concentrations of carcinogen with 10% calf serum. In serum-free culture, hyperplasia was induced in 2 weeks with one-tenth of the smallest concentration of benzo[a]pyrene that yielded this change in serum-containing medium. The cells of the hyperplastic epithelium in both serum-containing and serum-free culture exhibited ultrastructural changes described in carcinogenesis in vivo. In the absence of serum, squamous hyperplasia was uniformly seen, a feature that distinguished serum-free culture from culture in the presence of calf serum. No frank intraepithelial or invasive malignant lesions were produced in either medium. It was concluded that exposure of organ cultures to carcinogen in the absence of serum is the more promising method for bioassay because the response to carcinogen was more rapid, more sensitive, and more reproducible than that seen during exposure in media that contained serum.