Breast Microcalcifications Research Articles

Contrast-Enhanced Mammographic Features of In Situ and Invasive Ductal Carcinoma Manifesting Microcalcifications Only: Help to Predict Underestimation?

Simple SummaryMammography frequently detects the early breast cancers manifesting microcalcifications only. By means of mammographically guided vacuum-assisted core needle biopsy, noninvasive ductal carcinoma in situ (DCIS) is often diagnosed. Unfortunately, invasive components are occasionally embedded within the noninvasive cancer bed, which will alter the preoperative planning. Whether or not to perform single-step sentinel lymph node sampling is an area of controversy. In order to minimize the overtreatment or undertreatment, we retrospectively reviewed the enhanced features of DCIS and invasive ductal carcinomas (IDCs) on contrast-enhanced spectral mammograms (CESMs). The CESM is a modern technique enabling the provision of conventional mammograms and contrast-enhanced images. The results of our study revealed low DCIS upgrade of the unenhanced DCIS. Otherwise, DCIS tended to appear as nonmass and pure ground glass enhancements, and IDC tended to appear as mass and unpurified solid enhancement. The enhanced features allow distinguishing DCIS and IDC.Background: The contrast-enhanced mammographic features of ductal carcinoma in situ (DCIS) and invasive ductal carcinoma (IDC) manifesting microcalcifications only on mammograms were evaluated to determine whether they could predict IDC underestimation. Methods: We reviewed patients who underwent mammography-guided biopsy on suspicious breast microcalcifications only and received contrast-enhanced spectral mammography (CESM) within 2 weeks before the biopsy. Those patients who were proven to have cancers (DCIS or IDC) by biopsy and subsequently had surgical treatment in our hospital were included for analysis. The presence or absence, size, morphology and texture of enhancement on contrast-enhanced spectral mammography were reviewed by consensus of two radiologists. Results: A total of 49 patients were included for analysis. Forty patients (81.6%) showed enhancement, including 18 (45%) DCIS and 22 (55%) IDC patients. All nine unenhanced cancers were pure DCIS. Pure DCIS showed 72.22% nonmass enhancement and 83.33% pure ground glass enhancement. IDC showed more mass (72.2% vs. 27.8%) and solid enhancements (83.33% vs. 16.67%). The cancer and texture of enhancement were significantly different between pure DCIS and IDC, with moderate diagnostic performance for the former (p-value < 0.01, AUC = 0.66, sensitivity = 93%, specificity = 39%) and the latter (p-value < 0.01, AUC = 0.74, sensitivity = 65%, specificity = 83%). Otherwise, pure DCIS showed a significant difference in enhanced texture compared with upgraded IDC and IDC (p = 0.0226 and 0.0018, respectively). Conclusions: Nonmass and pure ground glass enhancements were closely related to pure DCIS, and cases showing mass and unpurified solid enhancements should be suspected as IDC. Unenhanced DCIS with microcalcifications only has a low DCIS upgrade rate. The CESM-enhanced features could feasibly predict IDC underestimation.

Diagnostic Performance of Artificial Intelligence-Based Computer-Aided Diagnosis for Breast Microcalcification on Mammography.

The present study evaluated the diagnostic performance of artificial intelligence-based computer-aided diagnosis (AI-CAD) compared to that of dedicated breast radiologists in characterizing suspicious microcalcification on mammography. We retrospectively analyzed 435 unilateral mammographies from 420 patients (286 benign; 149 malignant) undergoing biopsy for suspicious microcalcification from June 2003 to November 2019. Commercial AI-CAD was applied to the mammography images, and malignancy scores were calculated. Diagnostic performance was compared between radiologists and AI-CAD using the area under the receiving operator characteristics curve (AUC). The AUCs of radiologists and AI-CAD were not significantly different (0.722 vs. 0.745, p = 0.393). The AUCs of the adjusted category were 0.726, 0.744, and 0.756 with cutoffs of 2%, 10%, and 38.03% for AI-CAD, respectively, which were all significantly higher than those for radiologists alone (all p < 0.05). None of the 27 cases downgraded to category 3 with a cutoff of 2% were confirmed as malignant on pathological analysis, suggesting that unnecessary biopsies could be avoided. Our findings suggest that the diagnostic performance of AI-CAD in characterizing suspicious microcalcification on mammography was similar to that of the radiologists, indicating that it may aid in making clinical decisions regarding the treatment of breast microcalcification.

Role of micropure ultrasound in the assessment of grouped breast microcalcifications with or without a mass lesion

Background Grouped microcalcifications are the tiny grouping of the many calcifications seen on mammography. For pathological examination of mass-free clustered microcalcifications, a mammography is considered because its visualization by conventional ultrasound is limited. Therefore, MicroPure imaging improves its detection and visualization. Objective This study aimed to evaluate the accuracy of MicroPure ultrasound in the diagnosis and characterization of breast microcalcifications with or without a mass in women undergoing-guided biopsies by ultrasound. Patients and methods Thirty women who were referred for breast imaging and diagnosis from Surgery and Oncology Departments to the Radiodiagnosis Department at Tanta Cancer Center were included in this study. Results In terms of diagnosis, 26 (86.7%) women had a mass, in two women (6.7%), a mass was discovered during screening and in two women (6.7%), a mass was discovered during follow-up. There were 26 (86.7%) women with a clustered distribution and four (13.3%) with a segmental distribution. The mean value of microcalcification detected by mammography was 12.87±6.64. Eleven women (36.7%) had benign masses and 19 (63.3%) had malignant masses. In terms of histopathology, 11 (36.7%) women were ductal carcinoma in situ, eight (26.7%) were invasive ductal carcinoma, six (20%) with distal hyperplasia without atypia, two (6.7%) with atypical ductal hyperplasia, two (6.7%) with sclerosing adenosis and one (3.3%) with fibroadenosis. Regarding ultrasound B (US B) mode, 17 (56.7%) women were visible and 13 (43.4%) were not visible. Regarding MicroPure, 25 (83.3%) women were visible and five (16.7%) were not visible. The mean value of microcalcification detected by MicroPure and US B mode are 8.8±6.61 and 5.97±6.7, respectively. Conclusions MicroPure scanning is a hopeful ultrasound tool that might enhance the sensitivity for diagnosis clustered microcalcifications which are not combined with breast mass on mammography.

Predicting the pathological status of mammographic microcalcifications through a radiomics approach

Objective The study aimed to develop a machine learning (ML)-coupled interpretable radiomics signature to predict the pathological status of non-palpable suspicious breast microcalcifications (MCs).Methods We enrolled 463 digital mammographical view images from 260 consecutive patients detected with non-palpable MCs and BI-RADS scored at 4 (training cohort, n = 428; independent testing cohort, n = 35) in the First Affiliated Hospital of Nanjing Medical University between September 2010 and January 2019. Subsequently, 837 textures and 9 shape features were subsequently extracted from each view and finally selected by an XGBoost-embedded recursive feature elimination technique (RFE), followed by four machine learning-based classifiers to build the radiomics signature.Results Ten radiomic features constituted a malignancy-related signature for breast MCs as logistic regression (LR) and support vector machine (SVM) yielded better positive predictive value (PPV)/sensitivity (SE), 0.904 (95% CI, 0.865–0.949)/0.946 (95% CI, 0.929–0.977) and 0.891 (95% CI, 0.822–0.939)/0.939 (95% CI, 0.907–0.973) respectively, outperforming their negative predictive value (NPV)/specificity (SP) from 10-fold cross-validation (10FCV) of the training cohort. The optimal prognostic model was obtained by SVM with an area under the curve (AUC) of 0.906 (95% CI, 0.834–0.969) and accuracy (ACC) 0.787 (95% CI, 0.680–0.855) from 10FCV against AUC 0.810 (95% CI, 0.760–0.960) and ACC 0.800 from the testing cohort.Conclusion The proposed radiomics signature dependens on a set of ML-based advanced computational algorithms and is expected to identify pathologically cancerous cases from mammographically undecipherable MCs and thus offer prospective clinical diagnostic guidance.

Detection of Microcalcifications in Spiral Breast Computed Tomography with Photon-Counting Detector Is Feasible: A Specimen Study.

The primary objective of the study was to compare a spiral breast computed tomography system (SBCT) to digital breast tomosynthesis (DBT) for the detection of microcalcifications (MCs) in breast specimens. The secondary objective was to compare various reconstruction modes in SBCT. In total, 54 breast biopsy specimens were examined with mammography as a standard reference, with DBT, and with a dedicated SBCT containing a photon-counting detector. Three different reconstruction modes were applied for SBCT datasets (Recon1 = voxel size (0.15 mm)3, smooth kernel; Recon2 = voxel size (0.05 mm)3, smooth kernel; Recon3 = voxel size (0.05 mm)3, sharp kernel). Sensitivity and specificity of DBT and SBCT for the detection of suspicious MCs were analyzed, and the McNemar test was used for comparisons. Diagnostic confidence of the two readers (Likert Scale 1 = not confident; 5 = completely confident) was analyzed with ANOVA. Regarding detection of MCs, reader 1 had a higher sensitivity for DBT (94.3%) and Recon2 (94.9%) compared to Recon1 (88.5%; p < 0.05), while sensitivity for Recon3 was 92.4%. Respectively, reader 2 had a higher sensitivity for DBT (93.0%), Recon2 (92.4%), and Recon3 (93.0%) compared to Recon1 (86.0%; p < 0.05). Specificities ranged from 84.7–94.9% for both readers (p > 0.05). The diagnostic confidence of reader 1 was better with SBCT than with DBT (DBT 4.48 ± 0.88, Recon1 4.77 ± 0.66, Recon2 4.89 ± 0.44, and Recon3 4.75 ± 0.72; DBT vs. Recon1/2/3: p < 0.05), while reader 2 found no differences. Sensitivity and specificity for the detection of MCs in breast specimens is equal for DBT and SBCT when a small voxel size of (0.05 mm)3 is used with an equal or better diagnostic confidence for SBCT compared to DBT.

Computational FEM Model and Phantom Validation of Microwave Ablation for Segmental Microcalcifications in Breasts Using a Coaxial Double-Slot Antenna.

Introduction Cancer is the second leading cause of death worldwide. Breast cancer is the second most common cause of cancer-related mortality, accounting for 11.6% of the total number of deaths. The main treatments for this disease are surgical removal of the tumor, radiotherapy, and chemotherapy. Recently, different minimally invasive technologies have been applied (e.g., emission of electromagnetic waves, thermal and chemical means) to overcome the important side effects of these treatment modalities. The objective of this study was to develop and evaluate a predictive computational model of microwave ablation. Materials and Methods The predictive computational model of microwave ablation was constructed by means of a dual-slot coaxial antenna. The model was compared with an experiment performed using a breast phantom, which emulates the dielectric properties of breast tissue with segmental microcalcifications. The standing wave ratio (SWR) was obtained for both methods to make a comparison and determine the feasibility of applying electromagnetic ablation to premalignant lesions in breasts. Specifically, for the analysis of segmental microcalcifications, a breast phantom with segmental microcalcifications was developed and two computational models were performed under the same conditions (except for blood perfusion, which was excluded in one of the models). Results The SWR was obtained by triplicate experiments in the phantom, and the measurements had a difference of 0.191 between the minimum and maximum SWR values, implying a change of power reflection of 0.8%. The average of the three measurements was compared with the simulation that did not consider blood perfusion. The comparison yielded a change of 0.104, representing a 0.2% change in power reflection. Discussion. Both experimentation in phantom and simulations demonstrated that ablation therapy can be performed using this antenna. However, an additional optimization procedure is warranted to increase the efficiency of the antenna.

Suppression of artifacts in X-ray phase-contrast images retrieved by Fourier transform

Over the last two decades, the grating-based phase-contrast imaging has aroused the interest of a number of researchers. It could provide an access to three complementary signals simultaneously: the conventional absorption contrast, the differential phase contrast related to refraction of incident wave, and the dark-field contrast that relates to ultra small angle scattering in a sample. The grating-based phase-contrast signals have higher contrast sensitivity for some types of soft samples than the absorption signals. Dark-field signals have better diagnostic effects in the detection of lung tumors, pneumothorax and the identification of microcalcifications in breast. There are two main phase retrieval methods in grating-based X-ray phase-contrast imaging, i.e. phase stepping method and Fourier transform method. The phase signals retrieved by phase stepping is high precise and has low noise. But the sample suffers high dose due to at least three exposures. The phase signals retrieved by Fourier transform is low-dose due to the fact that only one image with sample is needed, but it is easily affected by artifacts when the size of the filtering window is too large. However, when the size of the filtering window is too small, the high-frequency information of the phase-contrast image will be lost, and the image will become blurred. A trade-off between definitions of the image and artifacts should be made. Since the phase-contrast signal and the dark-field signal of the sample are modulated by carrier fringes, the frequency spectrum of the detected image consists of many different harmonics. The artifacts in the retrieved signals originate from the spectrum aliasing between primary peak around zero spatial frequency and first-order harmonic peaks. Therefore, the subtraction between two images with phase difference can remove the primary peak, and the artifacts in the phase-contrast signals and dark-field signals will be suppressed. In order to further suppress the artifacts, we increase the frequency of carrier fringes, which results in a larger distance between first-order harmonic peaks in frequency domain. We finally attain artifact-free phase-contrast images and dark-field images while maintaining high definition of the images. The method proposed here is not only applicable to incoherent imaging system, but also to Talbot-Lau interferometer, and it would be useful in fast and low-dose X-ray phase-contrast and dark-field imaging.

Specimen Radiography for MRI-Guided Biopsies

Background: Specimen radiography is important for the biopsy of breast microcalcifications, and MRI is limited in the detection of microcalcifications. It is unknown whether or not the presence of microcalcifications on MRI-guided biopsies is significant. Purpose: To determine whether specimen radiography of MRI-guided biopsy samples provides any added benefits in tissue assessment. Materials and Methods: This is an IRB-approved, HIPPA-compliant retrospective review of MRI-guided biopsy reports whose tissue underwent specimen radiography from 2010 to 2017. Pathology reports were queried to compare samples with and without calcium and reviewed to determine if calcifications correlated with the lesion of interest. If there was a correlation, the original MRI was reviewed. Final pathology reports were also reviewed if excision was performed. Results: A total of 889 patients ages 22 - 85 were included with 140 (15.7%, 140/889) containing calcifications. Of 140 specimens, 119 (85.0%, 119/140) cases separated the calcifications. A total of 41 (34.5%, 41/119) were malignant or high-risk lesions/atypia of which 15 (36.6%, 15/41) showed a higher-grade lesion in the specimen containing calcium. Out of these 15, 4 (26.7%, 4/15) were pathologically associated with calcium; however, pathologic diagnosis was not dependent on the presence of calcifications. All 4 were high-risk lesions and none were malignancies. MRI in these cases showed three enhancing masses and one non-mass enhancement. None were upgraded at excision. Conclusion: The presence of microcalcifications on MRI-guided biopsies does not aid in tissue assessment and does not impact pathologic diagnosis. Specimen radiography provides no added benefits in the setting of MRI-guided biopsies.

Detection rate of breast malignancy of needle localization biopsy of breast microcalcification.

Objective:The current study aimed to retrospectively assess the cancer detection rate of needle localization biopsy of breast microcalcifications undetectable on sonography.Materials and Methods:Patients who underwent mammography-guided needle localization biopsy of breast microcalcifications undetectable on sonography from January 2005 to December 2017 were included in the study. Patients with incomplete medical records were excluded from the study. Patient mammograms were categorized using the Breast Imaging-Reporting and Data System (BI-RADS) assessment criteria. The percentages of benign and malignant lesions were determined by pathological examination of surgically recovered specimens. Correlation between preoperative imaging assessment and final diagnosis was investigated, and the complications associated with the procedures were recorded.Results:A total of 301 needle-localized biopsies were performed under mammographic guidance. The mean age of the patients was 58.18 ± 7.73 years. The overall ductal carcinoma in situ (DCIS) and cancer detection rate was 23.3%. The proportion of patients with BI-RADS 0 category and undergoing second mammography was higher in the DCIS and cancer group. A total of 227 patients did not undergo second mammography. Of these patients, 70 demonstrated BI-RADS 4 category, 34 were diagnosed with DCIS, and 5 were diagnosed with breast cancer during subsequent follow-up.Conclusion:Needle-localized excision of microcalcifications undetectable on sonography has high detection rate for early stage of breast cancer with low risk of associated complications. Regular mammography is a satisfactory follow-up tool for female patients with microcalcifications in the breasts. Additional studies should be performed to compare between needle-localized excision and vacuum-assisted breast biopsy.

EVALUATION OF DIAGNOSTIC ACCURACY OF ULTRASOUND-GUIDED SEMI-AUTOMATED CORE NEEDLE BIOPSY OF BREAST SUSPICIOUS MICROCALCIFICATIONS

Back ground: Breast microcalcifications are considered an early mammographic sign of breast cancer which are present with tiny bright spots of different morphology. In an ultrasound (US) image, the presence of microcalcifications within breast is an important indicator of malignancy.With an improved sonographic detection of microcalcification, ultrasound could be used for biopsy guidance for target sampling of tissue containing suspicious microcalcifications. If the biopsied lesions had a suspicious microcalcifications, specimen radiograph is mandatory to confirm the presence of microcalcifications in which a core containing any calcification on specimen radiography was defined as asuccessfully retrieved core. 14-G(Gauge) semi-automated core needle biopsy(CNB) device is less costly than other biopsy device. Methods:A prospective study was conducted on thirty female patients with thirty- two lesions their mean age was 48.53 years (age range, 21-75 years) radiologically classified as BIRADS (Breast Imaging Reporting and Data system) 4 (b, c) and5 from those referred to women imaging health unit in Radiology department for Sono- Mammography and those patients were referred from the inpatient wards and outpatient clinics of the surgery departments during the period from September 2018 till May 2019.All patients underwent conventional digital mammography and B-mode ultrasound examination and Ultrasound guided biopsy, specimen radiography then tissue samples were sent to histopathology department in which slide picture was done.Then (28) females done surgery and (2) patient still for follow up. Results: Radiological diagnosis of our lesions using BIRADS(Breast Imaging Reporting and Data system) categorical method were identified as two intermediate suspicious lesions (BIRADS 4B 6.3 %), twelve moderate suspicious lesions (BIRADS 4C 34.4 %), and 19 highly suggestive of malignancy lesions (BIRADS 5 59.4 %). Retrieval rate of calcifications according to specimen radiography were successfully retrieved in 30/32 lesions (93.8 %) and confirmed on specimen radiography .CNB( core needle biopsy) identified 28 lesions (87.5 %) malignant and four (12.5 %) benign lesions. Distribution of lesions according to pathological diagnosis in successful and fail retrieval group, the successful retrieval group comprised 26 malignant (22 IDC(Invasive ductal carcinoma), 2 DCIS(Ductal carcinoma Insitu), 2 ILC(Invasive Lobuler carcinoma) ) and four benign lesions.After CNB( core needle biopsy), 30 lesions underwent surgical excision and final pathology results were provided. All 24 IDC on 14-G semi-automated CNB (core needle biopsy) were confirmed to be IDC(Invasive ductal carcinoma) on final pathology. Of the two DCIS(Ductal carcinoma Insitu) on US(Ultrasound) guided CNB( core needle biopsy), one DCIS, were upgraded to IDC, the other one confirmed to be DCIS on final pathology,the two ILC were confirmed to be ILC on final pathology.Of the four benign lesions, the two ADH(Atypical Ductal hyperplasia) underwent surgical excision, finally diagnosed as DCIS,the other two benign lesions (fibroadenoma and fibroadenosis) their BIRADS assessments were category 4C and were recommended for follow-up.The overall accuracy of US-guided 14-G(Gauge) semi-automated CNB( core needle biopsy) was 90.0 % (27/32). Conclusion: The present study confirms the good retrieval rate (93.8%) of US-guided 14-G semi-automated CNB and diagnostic accuracy of (90.0%). Thus, this could be useful procedure for suspicious microcalcifications apparent on sonography.Specimen radiography proved to be a great help in deciding the accurate retrieval of microclcifications by core biopsy. It should be done in all cases of core biopsy for calcifications with marking of the core containing calcifications to be specially assessed by histopathology. We should try to do core biopsy before using VAB (Vaccum-Assisted Biobsy)for calcifications if well seen by ultrasound.

3D microcalcification detection using a color Doppler twinkling artifact with optimized transmit conditions: Preliminary results.

Mammography is the only method that has been proven to detect breast microcalcifications (MCs), but the sensitivity of mammography varies according to breast density. This paper proposes an ultrasound (US) color Doppler twinkling artifact (CDTA) method with optimized transmit conditions to identify breast MCs without ionizing radiation. The transmit conditions for US color Doppler imaging (CDI) were optimized to enhance the sensitivity of the twinkling artifact (TA) that arises from random scattering on rough surfaces of breast MCs. To validate the proposed breast MC detection method, a chicken breast phantom with MC particles (groups of particles <400 and <240 ) was fabricated and scanned by a digital mammography system and an US research platform by an L11-5v linear array probe with a three-dimensional (3D) motion tracking system. From the phantom experiment, the proposed 3D CDTA imaging method with optimized transmit conditions (i.e., a center frequency of 5.0MHz, an f-number of 1.3, and a peak negative pressure of 1.83MPa) successfully detected all 16 MC particles, comparable to detection with mammography. For a human breast surgical specimen in the ex vivo study, all 10 MC clusters, marked by a radiologist on the mammogram, were identified with the proposed 3D CDTA imaging method. In the phantom and ex vivo breast specimen studies, the proposed 3D CDTA imaging method successfully detected MCs, and the spatial localization was highly correlated with the mammogram results. These results indicate that the proposed 3D CDTA imaging method has great potential for the detection of MCs without ionizing radiation.

Classification of Mammographic Breast Microcalcifications Using a Deep Convolutional Neural Network: A BI-RADS-Based Approach.

Over 56,000 images of 268 mammograms from 94 patients were labeled to 3 classes according to the BI-RADS standard: "no microcalcifications" (BI-RADS 1), "probably benign microcalcifications" (BI-RADS 2/3), and "suspicious microcalcifications" (BI-RADS 4/5). Using the preprocessed images, a dCNN was trained and validated, generating 3 types of models: BI-RADS 4 cohort, BI-RADS 5 cohort, and BI-RADS 4 + 5 cohort. For the final validation of the trained dCNN models, a test data set consisting of 141 images of 51 mammograms from 26 patients labeled according to the corresponding BI-RADS classification from the radiological reports was applied. The performances of the dCNN models were evaluated, classifying each of the mammograms and computing the accuracy in comparison to the classification from the radiological reports. For visualization, probability maps of the classification were generated. The accuracy on the validation set after 130 epochs was 99.5% for the BI-RADS 4 cohort, 99.6% for the BI-RADS 5 cohort, and 98.1% for the BI-RADS 4 + 5 cohort. Confusion matrices of the "real-world" test data set for the 3 cohorts were generated where the radiological reports served as ground truth. The resulting accuracy was 39.0% for the BI-RADS 4 cohort, 80.9% for BI-RADS 5 cohort, and 76.6% for BI-RADS 4 + 5 cohort. The probability maps exhibited excellent image quality with correct classification of microcalcification distribution. The dCNNs can be trained to successfully classify microcalcifications on mammograms according to the BI-RADS classification system in order to act as a standardized quality control tool providing the expertise of a team of radiologists.

Digital breast tomosynthesis improves diagnostic accuracy of breast microcalcifications

AbstractDBT reconstructs high‐resolution tomographic images through multi‐angle scanning. We aimed at investigating the diagnostic value of DBT in breast microcalcifications of Asian women. The clinical characteristics and diagnostic accuracy of FFDM and DBT in 70 breast cancer patients were compared. 52 malignant lesions and 24 benign lesions were found in 76 breast microcalcifications. FFDM presented with high sensitivity and low specificity of 94.2% and 45.8%, while DBT presented with high sensitivity and medium specificity of 96.1% and 62.5%. The positive predictive value, negative predictive value, and diagnostic coincidence rate of DBT were higher than those of FFDM (84.7%, 88.2%, 85.5% vs. 79.0%, 78.5%, 78.9%, respectively). The AUC of DBT was significantly higher than that of FFDM (Z = 2.322, P = 0.020), at 0.700 ± 0.055 [0.584‐0.800] and 0.793 ± 0.052 [0.685‐0.878], respectively. Thus, we concluded that DBT improves the diagnostic accuracy and detection rate of microcalcifications in Asian women.

Discrimination of Benign and Malignant Lesions in Canine Mammary Tissue Samples Using Raman Spectroscopy: A Pilot Study.

Simple SummaryNeoplastic diseases are among the leading causes of death worldwide and constitute the main health problem in both human and veterinary medicine, particularly as the occurrence of the disease continues to increase. Comparative oncology is a quickly expanding field that examines both cancer risk and tumor development across species. Characterized by interdisciplinary collaboration, its goal is the improvement of both human and animal health. Canine neoplastic disease occurs spontaneously and has comparable clinical presentation and pathophysiology to corresponding human cancers. Since the nature of the disease is spontaneous, the complex interactions between tumor cells, tissues and the immune system can be better studied. Such relations are otherwise difficult to study in other experimental animal models. Raman spectroscopy has proved to be a suitable technique to detect and study breast microcalcifications. Raman spectroscopy is a specific and sensitive tool for identifying biomarkers of oncologic disease and also shows further potential in differentiating malignant and benign tumors, and these tumors from healthy tissue.Breast cancer is a health problem that affects individual life quality and the family system. It is the most frequent type of cancer in women, but men are also affected. As an integrative approach, comparative oncology offers an opportunity to learn more about natural cancers in different species. Methods based on Raman spectroscopy have shown significant potential in the study of the human breast through the fingerprinting of biological tissue, which provides valuable information that can be used to identify, characterize and discriminate structures in breast tissue, in both healthy and carcinogenic environments. One of the most important applications of Raman spectroscopy in medical diagnosis is the characterization of microcalcifications, which are highly important diagnostic indicators of breast tissue diseases. Raman spectroscopy has been used to analyze the chemical composition of microcalcifications. These occur in benign and malignant lesions in the human breast, and Raman helps to discriminate microcalcifications as type I and type II according to their composition. This paper demonstrates the recent progress in understanding how this vibrational technique can discriminate through the fingerprint regions of lesions in unstained histology sections from canine mammary glands.

Diagnosis of Breast Micro calcifications with Contrast Enhanced Digital Mammography and Histopathological Correlation

Abstract Backgroud: Microcalcificaitons constitute up to 31% of lesions detected at screening mammography. Ductal carcinoma in situ (DCIS) often presents with microcalcificaitons on mammography. Of all mammographically detected DCIS lesions, up to 79% manifest with microcalcifications only. However, not all microcalficications are associated with in situ or malignant disease. Aim of Study: To evaluate diagnostic value of contrast enhanced digital mammography in breast microcalcificaations. Methods: A retrospective study of 30 patients having mammographic suspicious breast calcifications under BIRADS 4, 5 and 6 categories. A high-energy and a low-energy digital mammograms were obtained and then images were subtracted, isolating the iodine signal in the region of angiogenesis. 1.5mg/kg of the contrast agent was injected by hand over a period of 1 minute at a rate of 3mL per second, followed by a saline solution flush. Breast compression was applied to reduce the thickness of the breast and images obtained exactly 2 min after contrast administration. Image acquisitions: Cranio-caudal and the medio-lateral oblique projections for the abnormal breast were obtained exactly 2 minutes after contrast injection followed by cranio-caudal and medio-lateral oblique projections of the normal breast. The lesions were analyzed by specialized radiologist for the presence, morphology, and pattern of enhancement. Finally, Correlation with histopatho-logical findings was done. Results: The sensitivity of CEDM in detecting malignant pathology with contrast uptake was 96%. Compared to sensi-tivity of 86.7% in FFDM. There were lesions of IDC in (69.2%), lesion of DCIS in (11.5%), lesion of IDCS & IDC in (11.5%) and lesions of ILC in (3.8%). Enhancement was also observed in 3/4 (75%) of the benign lesions. Conclusion: Dual-energy contrast-enhanced digital mam-mography is a useful technique in the diagnosis of underlying disease in mammographically detected breast microcalcifica-tions.

Abstract NG15: Investigation of relationships between breast cancer risk factors and bilateral mammographic breast density asymmetry among women undergoing diagnostic image-guided breast biopsies

Introduction: The etiology of breast cancer remains an area of ongoing investigation. Improving our understanding of factors associated with breast cancer development will strengthen the utility of risk prediction strategies and improve risk stratification. Mammographic breast density remains one of the most influential breast cancer risk factors, with 4 to 6 fold elevated risk observed among women with the highest compared to the lowest levels. These associations have consistently been observed irrespective of the method (visual or automated), used to quantify breast density, showing the robustness of the associations. An understudied breast cancer risk factor is the bilateral asymmetry of mammographic features, with prior studies suggesting that women with higher levels of breast asymmetry may be at elevated breast cancer risk. Increasingly, studies are recognizing the potential of bilateral breast asymmetry defined by mammographic breast density in determining breast cancer risk. Recently, Eriksson and colleagues highlighted a strong influence of breast asymmetry within a model they developed to predict short-term breast cancer risk among women attending breast screening. They observed that the contribution to the model of asymmetry in mammographic breast density, microcalcifications, and masses between the breasts was as substantial as the total number of microcalcifications and masses found within a mammogram, indicating that differences between the breasts may be an important risk marker. To help further understand associations between breast cancer risk factors and asymmetry of breast density, particularly among women at elevated risk for breast cancer, we evaluated risk factor relationships with the bilateral asymmetry of volumetric measures of global and local breast density across the spectrum of premalignant and invasive breast cancer diagnoses. Methods: This study included 882 women enrolled as part of the National Cancer Institute9s Breast Radiology Evaluation and Study of Tissues (BREAST)-Stamp Project (2007-2010). The BREAST-Stamp Project is a cross-sectional molecular epidemiologic study that aims to understand how novel breast density measures are related to breast cancer etiology. Women were enrolled if they were referred for diagnostic image-guided breast biopsy following an abnormal mammogram at the University of Vermont Medical Center, and had not previously been diagnosed or treated for cancer, undergone breast surgery within one year or received chemoprevention. Risk factor data were collected at study enrolment via interview and self-administered questionnaires. Breast density measures were estimated using Single X-ray Absorptiometry (SXA), a technique in which an SXA breast density phantom was affixed to the compression paddle of the mammography machine during routine mammography so that it was included in the X-ray field. It served as a reference standard to estimate volumetric breast density. Breast density was assessed using pre-biopsy craniocaudal full-field digital mammograms of both the ipsilateral (affected) and contralateral (unaffected) breast. Firstly, global density from each laterality was determined as percent fibroglandular volume (%FGV). Secondly, localized volumetric density measures were estimated following identification of the biopsy site on the ipsilateral pre-biopsy mammogram by the study radiologist and identifying the corresponding site on the contralateral mammogram. The SXA estimated %FGV in a perilesional volume twice the size of, but excluding, the biopsy target, centered at the biopsy site. Previous estimates of reproducibility for the SXA test phantoms demonstrated a repeatability SD of 2%, with a ±2% accuracy for the entire thickness and density ranges. Breast density asymmetry was defined as an absolute bilateral difference in measures when subtracting the breast density measures of the contralateral breast from the ipsilateral measures. Spearman9s correlations (rho) examined associations between breast density measures of the left and right breasts. To determine relationships between breast cancer risk factors (defined as categorical variables) and measures of bilateral breast density asymmetry, analysis of covariance (ANCOVA) models (PROC GLM) were used. Analyses were conducted at the per woman level using SAS. Probability values of Citation Format: Maeve Mullooly, Shaoqi Fan, Ruth M. Pfeiffer, Brian Sprague, Pamela M. Vacek, Donald L. Weaver, John A. Shepherd, Amir Pasha Mahmoudzadeh, Jeff Wang, Serghei Malkov, Jason M. Johnson, Sally D. Herschorn, Gretchen L. Gierach. Investigation of relationships between breast cancer risk factors and bilateral mammographic breast density asymmetry among women undergoing diagnostic image-guided breast biopsies [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr NG15.

Abstract 5276: Secretory pathway calcium ATPase 2 (SPCA2) promotes cell survival and chemoresistance in receptor positive breast cancer cells

Abstract Introduction: Dysregulation of the Ca2+ toolkit has profound consequences for tumor growth and metastasis, raising hopes for novel avenues of therapeutic intervention. The Secretory Pathway Ca2+-ATPase Isoform 2 (SPCA2) has a dual function in sequestering Ca2+ and Mn2+ into secretory stores as well as eliciting their entry into cells by activating plasma membrane ion channels. We have previously showed that SPCA2 is implicated in breast cancer progression, microcalcifications, epithelial-mesenchymal transition, and metastasis. Methods and Results: High SPCA2 expression was associated with poor overall survival in receptor positive breast cancer patients (n=548, P =8.7e-0.6; KM Plotter). Depletion of SPCA2 (SPCA2 KD) in MCF-7 cells significantly reduced cell proliferation (P &amp;lt; 0.05). Analysis of the cell cycle (PI staining, Flow cytometry) revealed a significant increase (P &amp;lt; 0.05) in the number of cells in the G0/G1 phase, and significant increase in apoptosis (via TUNEL assay) (P &amp;lt; 0.05) in SPCA2 KD cells compared to control. Microarray analysis showed the p53 pathway is the most significantly upregulated pathway (P &amp;lt; 0.05) in SPCA2 KD cells compared to control. P53 knockout abrogated SPCA2 mediated cell cycle arrest and apoptosis. Depletion of SPCA2 increased reactive oxygen species (ROS, measured by DCFDA dye and MitoSox Red using flow cytometry) leading to DNA damage (confocal microscopy), and activation of ATM/ATR pathway (Immunoblotting, P &amp;lt; 0.05). As a consequence, SPCA2 KD cells showed specific hypersensitivity to DNA damaging agents used in breast cancer therapy (carboplatin, radiation, and doxorubicin) (determined by MTT assay) (P &amp;lt; 0.05). Thus SPCA2 is a novel molecular target in breast cancer therapy. Curcumin is currently being evaluated in clinical trials of breast cancer. We show that curcumin causes dose-dependence inhibition of SPCA2 expression (P &amp;lt; 0.05) in multiple receptor-positive cell lines, and that forced overexpression of SPCA2 decreased curcumin cytotoxicity. Interestingly, we did not observe these results with SPCA1, the housekeeping SPCA isoform, revealing isoform-specific function of Golgi calcium pumps in breast cancer subtypes. Conclusion: These novel findings point to a causal link between SPCA2, breast cancer progression, and chemoresistance to DNA damaging agents. Decrease of SPCA2 expression by curcumin may have therapeutic potential in treating receptor positive breast cancer. Citation Format: Monish Ram Makena, Donna Dang, Allatah Mekile, Phillip Buckhaults, Rajini Rao. Secretory pathway calcium ATPase 2 (SPCA2) promotes cell survival and chemoresistance in receptor positive breast cancer cells [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 5276.

Omega-3 fatty acids in pathological calcification and bone health.

Omega-3 fatty acids (ω-3FAs) such as Docosahexaenoic acid (DHA) and Eicosapentanoic acid (EPA), are active ingredient of fish oil, which have larger health benefits against various diseases including cardiovascular, neurodegenerative, cancers and bone diseases. Substantial studies documented a preventive role of omega-3 fatty acids in pathological calcification like vascular calcification and microcalcification in cancer tissues. In parallel, these fatty acids improve bone quality probably by preventing bone decay and augmenting bone mineralization. This study also addresses that the functions of ω-3FAs not only depend on tissue types, but also work through different molecular mechanisms for preventing pathological calcification in various tissues and improving bone health. PRACTICAL APPLICATIONS: Practical applications of the current study are to improve the knowledge about the supplementation of omega-3 fatty acids. This study infers that supplementation of omega-3 fatty acids aids in bone preservation in elder females at the risk of osteoporosis and also, on the contrary, omega-3 fatty acids interfere with pathological calcification of vascular cells and cancer cells. Omega-3 supplementation should be given to the cardiac patients because of its cardio protective role. In line with this, omega-3 supplementation should be included with chemotherapy for cancer patients as it can prevent osteoblastic potential of breast cancer patients, responsible for pathological mineralization, and blocks off target toxicities. Administration of omega-3 fatty acid with chemotherapy will not only improve survival of cancer patients, but also improve the bone quality. Thus, this study allows a better understanding on omega-3 fatty acids in combating pathological complications such as osteoporosis, vascular calcification, and breast microcalcification.

BMP-2 Variants in Breast Epithelial to Mesenchymal Transition and Microcalcifications Origin.

This study aims to investigate the possible different roles of the BMP-2 variants, cytoplasmic and nuclear variant, in both epithelial to mesenchymal transition and in microcalcifications origin in human breast cancers. To this end, the in situ expression of cytoplasmic and nuclear BMP-2 was associated with the expression of the main epithelial to mesenchymal transition biomarkers (e-cadherin and vimentin) and molecules involved in bone metabolisms (RUNX2, RANKL, SDF-1) by immunohistochemistry. In addition, the expression of cytoplasmic and nuclear BMP-2 was associated with the presence of microcalcifications. Our data showed a significant association among the number of cytoplasmic BMP-2-positive cells and the number of both vimentin (positive association) and e-cadherin (negative association) positive breast cells. Conversely, no associations were found concerning the nuclear BMP-2-positive breast cells. Surprisingly, the opposite result was obtained by analyzing the variants of BMP-2 and both the expression of RANKL and SDF-1 and the presence of microcalcifications. Specifically, the presence of microcalcifications was related to the expression of nuclear BMP-2 variant rather than the cytoplasmic one, as well as a strong association between the number of nuclear BMP-2 and the expression of the main breast osteoblast-like cells (BOLCs) biomarkers. To further corroborate these data, an in vitro experiment for demonstrating the co-expression of nBMP-2 and RANKL or vimentin or SDF-1 in breast cancer cells that acquire the capability to produce microcalcifications was developed. These investigations confirmed the association between the nBMP-2 expression and both RANKL and SDF-1. The data supports the idea that whilst cytoplasmic BMP-2 can be involved in epithelial to mesenchymal transition phenomenon, the nuclear variant is related to the essential mechanisms for the formation of breast microcalcifications. In conclusion, from these experimental and translational perspectives, the complexity of BMP-2 signaling will require a detailed understanding of the involvement of specific BMP-2 variants in breast cancers.

P123: Preliminary evaluation of raman spectroscopy to detect calcifications in excised breast tissue: A prelude to the first in vivo whole breast study

Introduction: Mammographic screening through the NHS Breast Screening Programme (NHSBSP) has limitations, including false positivity (9-13%). Adjunct techniques would support clinical decision making and reduce unnecessary biopsy. Raman spectroscopy is an analytical method that is non-invasive, non-destructive and provides chemical information to distinguish between breast micro-calcifications: Type I (benign) and Type II (malignant). Baker et al (2010) demonstrated that the level of carbonate substitution in type II calcifications can help provide cancer grade.