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In this paper, we present the novel Deep-MEG approach in which image-based representations of magnetoencephalography (MEG) data are combined with ensemble classifiers based on deep convolutional neural networks. For the scope of predicting the early signs of Alzheimer’s disease (AD), functional connectivity (FC) measures between the brain bio-magnetic signals originated from spatially separated brain regions are used as MEG data representations for the analysis. After stacking the FC indicators relative to different frequency bands into multiple images, a deep transfer learning model is used to extract different sets of deep features and to derive improved classification ensembles. The proposed Deep-MEG architectures were tested on a set of resting-state MEG recordings and their corresponding magnetic resonance imaging scans, from a longitudinal study involving 87 subjects. Accuracy values of 89% and 87% were obtained, respectively, for the early prediction of AD conversion in a sample of 54 mild cognitive impairment subjects and in a sample of 87 subjects, including 33 healthy controls. These results indicate that the proposed Deep-MEG approach is a powerful tool for detecting early alterations in the spectral–temporal connectivity profiles and in their spatial relationships.

The incremented uptake provided by time-lapse microscopy in Organ-on-a-Chip (OoC) devices allowed increased attention to the dynamics of the co-cultured systems. However, the amount of information stored in long-time experiments may constitute a serious bottleneck of the experimental pipeline. Forward long-term prediction of cell trajectories may reduce the spatial–temporal burden of video sequences storage. Cell trajectory prediction becomes crucial especially to increase the trustworthiness in software tools designed to conduct a massive analysis of cell behavior under chemical stimuli. To address this task, we transpose here the exploitation of the presence of “social forces” from the human to the cellular level for motion prediction at microscale by adapting the potential of Social Generative Adversarial Network predictors to cell motility. To demonstrate the effectiveness of the approach, we consider here two case studies: one related to PC-3 prostate cancer cells cultured in 2D Petri dishes under control and treated conditions and one related to an OoC experiment of tumor-immune interaction in fibrosarcoma cells. The goodness of the proposed strategy has been verified by successfully comparing the distributions of common descriptors (kinematic descriptors and mean interaction time for the two scenarios respectively) from the trajectories obtained by video analysis and the predicted counterparts.

Organs On a Chip (OOCs) represent a sophisticated approach for exploring biological mechanisms and developing therapeutic agents. In conjunction with high-quality time-lapse microscopy (TLM), OOCs allow for the visualization of reconstituted complex biological processes, such as multi-cell-type migration and cell–cell interactions. In this context, increasing the frame rate is desirable to reconstruct accurately cell-interaction dynamics. However, a trade-off between high resolution and carried information content is required to reduce the overall data volume. Moreover, high frame rates increase photobleaching and phototoxicity. As a possible solution for these problems, we report a new hybrid-imaging paradigm based on the integration of OOC/TLMs with a Multi-scale Generative Adversarial Network (GAN) predicting interleaved video frames with the aim to provide high-throughput videos. We tested the performance of the predictive capability of GAN on synthetic videos, as well as on real OOC experiments dealing with tumor–immune cell interactions. The proposed approach offers the possibility to acquire a reduced number of high-quality TLM images without any major loss of information on the phenomena under investigation.

We describe a novel method to achieve a universal, massive, and fully automated analysis of cell motility behaviours, starting from time-lapse microscopy images. The approach was inspired by the recent successes in application of machine learning for style recognition in paintings and artistic style transfer. The originality of the method relies i) on the generation of atlas from the collection of single-cell trajectories in order to visually encode the multiple descriptors of cell motility, and ii) on the application of pre-trained Deep Learning Convolutional Neural Network architecture in order to extract relevant features to be used for classification tasks from this visual atlas. Validation tests were conducted on two different cell motility scenarios: 1) a 3D biomimetic gels of immune cells, co-cultured with breast cancer cells in organ-on-chip devices, upon treatment with an immunotherapy drug; 2) Petri dishes of clustered prostate cancer cells, upon treatment with a chemotherapy drug. For each scenario, single-cell trajectories are very accurately classified according to the presence or not of the drugs. This original approach demonstrates the existence of universal features in cell motility (a so called “motility style”) which are identified by the DL approach in the rationale of discovering the unknown message in cell trajectories.

Cell-cell interactions are an observable manifestation of underlying complex biological processes occurring in response to diversified biochemical stimuli. Recent experiments with microfluidic devices and live cell imaging show that it is possible to characterize cell kinematics via computerized algorithms and unravel the effects of targeted therapies. We study the influence of spatial and temporal resolutions of time-lapse videos on motility and interaction descriptors with computational models that mimic the interaction dynamics among cells. We show that the experimental set-up of time-lapse microscopy has a direct impact on the cell tracking algorithm and on the derived numerical descriptors. We also show that, when comparing kinematic descriptors in two diverse experimental conditions, too low resolutions may alter the descriptors’ discriminative power, and so the statistical significance of the difference between the two compared distributions. The conclusions derived from the computational models were experimentally confirmed by a series of video-microscopy acquisitions of co-cultures of unlabelled human cancer and immune cells embedded in 3D collagen gels within microfluidic devices. We argue that the experimental protocol of acquisition should be adapted to the specific kind of analysis involved and to the chosen descriptors in order to derive reliable conclusions and avoid biasing the interpretation of results.

The dynamic contrast-enhanced MR imaging plays a crucial role in evaluating the effectiveness of neoadjuvant chemotherapy (NAC) even since its early stage through the prediction of the final pathological complete response (pCR). In this study, we proposed a transfer learning approach to predict if a patient achieved pCR (pCR) or did not (non-pCR) by exploiting, separately or in combination, pre-treatment and early-treatment exams from I-SPY1 TRIAL public database. First, low-level features, i.e., related to local structure of the image, were automatically extracted by a pre-trained convolutional neural network (CNN) overcoming manual feature extraction. Next, an optimal set of most stable features was detected and then used to design an SVM classifier. A first subset of patients, called fine-tuning dataset (30 pCR; 78 non-pCR), was used to perform the optimal choice of features. A second subset not involved in the feature selection process was employed as an independent test (7 pCR; 19 non-pCR) to validate the model. By combining the optimal features extracted from both pre-treatment and early-treatment exams with some clinical features, i.e., ER, PgR, HER2 and molecular subtype, an accuracy of 91.4% and 92.3%, and an AUC value of 0.93 and 0.90, were returned on the fine-tuning dataset and the independent test, respectively. Overall, the low-level CNN features have an important role in the early evaluation of the NAC efficacy by predicting pCR. The proposed model represents a first effort towards the development of a clinical support tool for an early prediction of pCR to NAC.

Anemia; Microfluídica; Enfermedad de células falciformes Anèmia; Microfluídica; Malaltia de cèl·lules falciformes Anaemia; Microfluidics; Sickle cell disease Combining microfluidics technology with machine learning represents an innovative approach to conduct massive quantitative cell behavior study and implement smart decision-making systems in support of clinical diagnostics. The spleen plays a key-role in rare hereditary hemolytic anemia (RHHA), being the organ responsible for the premature removal of defective red blood cells (RBCs). The goal is to adapt the physiological spleen filtering strategy for in vitro study and monitoring of blood diseases through RBCs shape analysis. Then, a microfluidic device mimicking the slits of the spleen red pulp area and video data analysis are combined for the characterization of RBCs in RHHA. This microfluidic unit is designed to evaluate RBC deformability by maintaining them fixed in planar orientation, allowing the visual inspection of RBC’s capacity to restore their original shape after crossing microconstrictions. Then, two cooperative learning approaches are used for the analysis: the majority voting scheme, in which the most voted label for all the cell images is the class assigned to the entire video; and the maximum sum of scores to decide the maximally scored class to assign. The proposed platform shows the capability to discriminate healthy controls and patients with an average efficiency of 91%, but also to distinguish between RHHA subtypes, with an efficiency of 82%. This work was supported by Networking Biomedical Research Center (CIBER), Spain. CIBER is an initiative funded by the VI National R&D&i Plan 2008–2011, Iniciativa Ingenio 2010, Consolider Program, CIBER Actions, and the Instituto de Salud Carlos III (RD16/0006/0012), with the support of the European Regional Development Fund. This work was funded by the European Commission H2020-MSCA-ITN-2019, Grant Agreement N860436, “EVIDENCE” and by the CERCA Programme and by the Commission for Universities and Research of the Department of Innovation, Universities, and Enterprise of the Generalitat de Catalunya (2017 SGR 1079), it has been developed in the context of AdvanceCat with the support of ACCIÓ (Catalonia Trade and Investment; Generalitat de Catalunya) under the Catalonian ERDF operational program (European Regional Development Fund) 2014–2020.

Simple Summary An early prediction of Breast Cancer Recurrence (BCR) for patients undergoing neoadjuvant chemotherapy (NACT) could better guide clinicians in the identification of the most suitable combination treatments for individual patient scenarios. We proposed a transfer learning approach to give an early prediction of three-year BCR for patients undergoing NACT, using DCE-MRI exams from I-SPY1 TRIAL and BREAST-MRI-NACT-Pilot public databases. Because no technical expertise is required in the extraction of meaningful features from images, the predictive model qualifies as a user-friendly tool for any medical expert in support of therapeutic choices. Only pre-treatment and early-treatment MRI examinations were analyzed to allow for potential therapy changes at a very early stage of treatment. We tested the strength of the model on an independent test. The best predictive performances (accuracy of 85.2%, sensitivity of 84.6%, and AUC of 0.83) were achieved by combining the extracted features with some clinical factors: age, ER, PgR, HER2+. Abstract Cancer treatment planning benefits from an accurate early prediction of the treatment efficacy. The goal of this study is to give an early prediction of three-year Breast Cancer Recurrence (BCR) for patients who underwent neoadjuvant chemotherapy. We addressed the task from a new perspective based on transfer learning applied to pre-treatment and early-treatment DCE-MRI scans. Firstly, low-level features were automatically extracted from MR images using a pre-trained Convolutional Neural Network (CNN) architecture without human intervention. Subsequently, the prediction model was built with an optimal subset of CNN features and evaluated on two sets of patients from I-SPY1 TRIAL and BREAST-MRI-NACT-Pilot public databases: a fine-tuning dataset (70 not recurrent and 26 recurrent cases), which was primarily used to find the optimal subset of CNN features, and an independent test (45 not recurrent and 17 recurrent cases), whose patients had not been involved in the feature selection process. The best results were achieved when the optimal CNN features were augmented by four clinical variables (age, ER, PgR, HER2+), reaching an accuracy of 91.7% and 85.2%, a sensitivity of 80.8% and 84.6%, a specificity of 95.7% and 85.4%, and an AUC value of 0.93 and 0.83 on the fine-tuning dataset and the independent test, respectively. Finally, the CNN features extracted from pre-treatment and early-treatment exams were revealed to be strong predictors of BCR.

Combining microfluidics technology with machine learning represents an innovative approach to conduct massive quantitative cell behavior study and implement smart decision-making systems in support of clinical diagnostics. The spleen plays a key-role in rare hereditary hemolytic anemia (RHHA), being the organ responsible for the premature removal of defective red blood cells (RBCs). The goal is to adapt the physiological spleen filtering strategy for in vitro study and monitoring of blood diseases through RBCs shape analysis. Then, a microfluidic spleen-like filtering unit and video data analysis are combined for the characterization of RBCs in RHHA. A filtering unit is designed to measure deformability by maintaining fixed the RBC orientation to study its capacity of restoring the original shape after crossing microconstrictions. Two cooperative learning approaches are used for the analysis: the majority voting scheme, in which the most voted label for all the cell images is the class assigned to the entire video; and the maximum sum of scores to decide the maximally scored class to assign. The proposed platform shows the capability to discriminate healthy controls and patients with an average efficiency of 91%, but also to distinguish between RHHA subtypes, with an efficiency of 82%.

Oncoimmunology represents a biomedical research discipline coined to study the roles of immune system in cancer progression with the aim of discovering novel strategies to arm it against the malignancy. Infiltration of immune cells within the tumor microenvironment is an early event that results in the establishment of a dynamic cross-talk. Here, immune cells sense antigenic cues to mount a specific anti-tumor response while cancer cells emanate inhibitory signals to dampen it. Animals models have led to giant steps in this research context, and several tools to investigate the effect of immune infiltration in the tumor microenvironment are currently available. However, the use of animals represents a challenge due to ethical issues and long duration of experiments. Organs-on-chip are innovative tools not only to study how cells derived from different organs interact with each other, but also to investigate on the crosstalk between immune cells and different types of cancer cells. In this review, we describe the state-of-the-art of microfluidics and the impact of OOC in the field of oncoimmunology underlining the importance of this system in the advancements on the complexity of tumor microenvironment.

The blood-brain barrier (BBB) is a dynamic cellular barrier that regulates brain nutrient supply, waste efflux, and paracellular diffusion through specialized junctional complexes. Finding a system to mimic and monitor BBB integrity (i.e., to be able to assess the effect of certain compounds on opening or closing the barrier) is of vital importance in several pathologies. This work aims to overcome some limitations of current barrier integrity measuring techniques thanks to a multi-layer microfluidic platform with integrated electrodes and Multi-frequency Trans-Endothelial Electrical Resistance (MTEER) in synergy with machine learning algorithms. MTEER measurements are performed across the barrier in a range of frequencies up to 10 MHz highlighting the presence of information on different frequency ranges. Results show that the proposed platform can detect barrier formation, opening, and regeneration afterwards, correlating with the results obtained from immunostaining of junctional complexes. This model presents novel techniques for a future biological barrier in-vitro studies that could potentially help on elucidating barrier opening or sealing on treatments with different drugs., This work was supported by Networking Biomedical Research Center (CIBER), Spain. CIBER is an initiative funded by the VI National R&D&i Plan 2008–2011, Iniciativa Ingenio 2010, Consolider Program, CIBER Actions, and the Instituto de Salud Carlos III (RD16/0006/0012), with the support of the European Regional Development Fund. This work was funded by the CERCA Programme and by the Commission for Universities and Research of the Department of Innovation, Universities, and Enterprise of the Generalitat de Catalunya (2017 SGR 1079). This work has been developed in the context of AdvanceCat with the support of ACCIÓ (Catalonia Trade and Investment; Generalitat de Catalunya) under the Catalonian ERDF operational program (European Regional Development Fund) 2014–2020. This work was funded by the Spanish Ministry of Economy and Competitiveness (MINECO) through the projects MINDS (Proyectos I + D Excelencia + FEDER) TEC2015-70104-P, CTQ2016-75870-P, BIOBOT (Programa Explora Ciencia / Tecnología), TEC2015- 72718-EXP and RTI2018-097038-B-C21.

Background: A Critical Shoulder Angle (CSA), evaluated on plain radiographs, greater than 35&deg, is considered predictive of rotator cuff tears. The present prospective comparative study aimed, firstly, to develop a formula to calculate the amount of acromion that should be resected performing a lateral acromioplasty and, secondly, verify whether lateral acromioplasty to reduce the CSA associated with arthroscopic cuff repair decreased the rate of recurrence of the tears, and impacted favorably on clinical postoperative outcomes. Methods: Patients undergoing arthroscopic rotator cuff repair (RCR) for rotator cuff tears with a CSA greater than 35&deg, were included in this study and divided into two groups, based on whether the CSA had been reduced by arthroscopic resection of the lateral portion of the acromion. A new mathematical formula was developed in order to quantify the amount of bone to be resected while performing the lateral acromioplasty. Patients with traumatic tears, previous surgery, osteoarthritis or plain radiographs, not classified as A1 according to Suter-Henninger, were excluded. Clinical and radiographic outcomes were assessed at a minimum of 2 years of follow-up considering the tear size. Results: 289 patients were included in this study. Thirty-seven were lost to follow-up. Group A (Lateral acromioplasty) patients included: 38 small tears, 30 medium tears, 28 large tears and 22 massive tears, Group B (control group) was composed of 40 small tears, 30 medium tears, 30 large tears and 23 massive tears. The Constants Score value and retear Rate were, respectively, significant higher (p = 0.007 and p = 0.004) and lower (p = 0.029 and p = 0.028) in Group A, both in the Small-and Medium-size subgroups. No complications were outlined. The mediolateral width of the acromion was reduced, according to the preoperatively calculated measure. Conclusion: Arthroscopic lateral acromioplasty decreased the CSA within the favorable range (30&deg, &ndash, 35&deg, ) in all patients treated, resecting the amount of bone predicted by the mathematical formula. Lateral acromioplasty is a safe and reproducible technique which may prevent recurrence of rotator cuff tears in patients with small and medium lesions. Level of evidence: II.

Localization of malignant sites of asymmetry in mammograms.Combination of Tabar masking procedures.Use of correlation-based structural similarity descriptors.Database-independent validation strategy. Background and objectivesThe analysis of patterns of asymmetry between the left and right mammograms of a patient can provide meaningful insights into the presence of an underlying tumor in its early stage. However, the identification of breast cancer by investigating bilateral asymmetry is difficult to perform due to the indistinct and borderline nature of the asymmetric signs as they appear on mammograms. MethodsIn this study, to increase the positive-predictive value of asymmetry in mammographic screening, a novel computerized approach for the automatic localization of malignant sites of asymmetry in mammograms is proposed. The sites of anatomical correspondence between the right and left regions of each radiographic projection were extracted by means of two bilateral masking procedures, inspired by radiologists criteria in interpreting mammograms and based on the use of detected landmarking structures. Relative variations of spatial patterns of intensity values and of orientations of directional components within each site were quantified by combining multidirectional Gabor filters and indices of structural similarity. The localization of the sites of malignant asymmetry was performed by coupling two quadratic discriminant analysis classifiers, one for each masking procedure, that assigned the likelihood of malignancy to each site of correspondence. ResultsThe performance of the proposed method was assessed on 94 mammographic images from two publicly available databases and containing at least one asymmetric site. Sensitivity, specificity and balanced accuracy levels of 0.83 (0.09), 0.75 (0.06), and 0.79 (0.04), respectively were obtained in the classification of malignant asymmetric sites vs benign/normal sites using cross-validation. In addition, a further blind test on a dataset of Full Field Digital Mammograms achieved levels of sensitivity, specificity, and balanced accuracy of 0.86, 0.65, and 0.75, respectively. ConclusionsThe achieved performance indicates that the proposed system is effective in localizing sites of malignant asymmetry and it is expected to improve computer-aided diagnosis of breast cancer.

Understanding the interactions between immune and cancer cells occurring within the tumor microenvironment is a prerequisite for successful and personalized anti-cancer therapies. Microfluidic devices, coupled to advanced microscopy systems and automated analytical tools, can represent an innovative approach for high-throughput investigations on immune cell-cancer interactions. In order to study such interactions and to evaluate how therapeutic agents can affect this crosstalk, we employed two ad hoc fabricated microfluidic platforms reproducing advanced 2D or 3D tumor immune microenvironments. In the first type of chip, we confronted the capacity of tumor cells embedded in Matrigel containing one drug or Matrigel containing a combination of two drugs to attract differentially immune cells, by fluorescence microscopy analyses. In the second chip, we investigated the migratory/interaction response of naive immune cells to danger signals emanated from tumor cells treated with an immunogenic drug, by time-lapse microscopy and automated tracking analysis. We demonstrate that microfluidic platforms and their associated high-throughput computed analyses can represent versatile and smart systems to: (i) monitor and quantify the recruitment and interactions of the immune cells with cancer in a controlled environment, (ii) evaluate the immunogenic effects of anti-cancer therapeutic agents and (iii) evaluate the immunogenic efficacy of combinatorial regimens with respect to single agents.

Background and objective: Full-field digital mammography (FFDM) is the predominant breast cancer screening exam used. However, with the emergence of digital breast tomosynthesis (DBT) the radiologists could improve early recognition of breast cancer signs. In this scenario, the detection of architectural distortion (AD) is still a challenging task. ADs are very subtle contraction of the breast parenchyma that could represent the earliest manifestation of cancer, assessing at present 50% of missed cases. Methods: This paper proposes a new paradigm to detect AD in DBT exams by a cross-cutting approach exploiting the 3-dimensionality of the imaging modality. After locating AD candidates in each DBT slice, the suspicious spots are tracked in cross-slice direction and then characterized in terms of neighboring texture. In this approach, which mimics radiologist's scrolling down over zoomed slices, we reduce the amount of uninformative signs collected in DBT exams by preserving the large variability of AD appearance. Results: Using 37 sets of DBT slices containing at least one AD locus indicated by a radiologist, the proposed methodology reaches an AUC of 0.84, with only one false negative exam at sensitivity of 0.9. Conclusions: The results show that the proposed algorithm can be a promising tool for the automatic detection of AD locii. Future work will address the extension of the dataset of DBT slices as well the improvement of algorithm performance toward the application in the clinical practice.

Objective: The ability of cells to collectively move is essential in various biological contexts including cancer metastasis. In this paper, we propose an automatic video analysis tool to correlate the cell movement inhibition with replication block induced by dose-dependent chemotherapy administration. Methods: The novel approach combines individual and collective cell kinematic analysis performed over time-lapse microscopy video frames. Cells are first localized and tracked, and then kinematic descriptors are extracted for each track. Selective track identification is performed assuming diversified cell roles within the same cluster (spontaneously forming groups of cells), and finally individual results are grouped exploiting consensus of coordinated motility within cell clusters. Results: Recognition performance of three different experimental conditions (no drug, 0.5–5 μ M merged in the same condition, and 50 μ M) reached an average accuracy value of 88% over 958 different tracks collected in 36 clusters of diverse dimensions in eight independent experiments. Conclusion: An extensive application of this methodology could give a different point of view of the cancer mechanisms.

A major challenge in cancer research is the complexity of the tumor microenvironment and the necessity to take into account the host immunological setting. An innovative way to address these problems is to exploit the emerging technology of organ-on-chip to achieve co-cultures in microfluidic devises that recapitulate ex vivo the tumor ecosystem. We generated tumors-on-chip integrating four cell populations (cancer, immune, endothelial cells and fibroblasts) in a 3D collagen matrix, reconstituting HER2 breast cancer ecosystem. By time-lapse microscopy, differential live staining and a novel automated analysis method, we visualized, and quantified the complex dynamics of this ecosystem, in absence or in presence of the drug trastuzumab (Herceptin), a targeted antibody therapy directed against the HER2 receptor. We uncovered the capacity of the drug trastuzumab to specifically promote long cancer-immune interactions (≳ 60 min), recapitulating an anti-tumoral ADCC (antibody-dependent cell-mediated cytotoxicity) immune response. Cancer-associated fibroblasts (CAFs) on the contrary inhibited cancer-immune interactions, antagonizing the action of the trastuzumab. These observations constitute a proof-of-concept that tumors-on-chip are novel powerful platforms that can be exploited to study ex vivo immunocompetent tumor microenvironments, to characterize ecosystem-level responses to anti-cancer drugs, and to dissect the roles of the various cellular components of the stroma.

This paper proposes a method to reduce the number of false-positives (FP) in a computer-aided detection (CAD) scheme for automated detection of architectural distortion (AD) in digital mammography. AD is a subtle contraction of breast parenchyma that may represent an early sign of breast cancer. Due to its subtlety and variability, AD is more difficult to detect compared to microcalcifications and masses, and is commonly found in retrospective evaluations of false-negative mammograms. Several computer-based systems have been proposed for automated detection of AD in breast images. The usual approach is automatically detect possible sites of AD in a mammographic image (segmentation step) and then use a classifier to eliminate the false-positives and identify the suspicious regions (classification step). This paper focus on the optimization of the segmentation step to reduce the number of FPs that is used as input to the classifier. The proposal is to use statistical measurements to score the segmented regions and then apply a threshold to select a small quantity of regions that should be submitted to the classification step, improving the detection performance of a CAD scheme. We evaluated 12 image features to score and select suspicious regions of 74 clinical Full-Field Digital Mammography (FFDM). All images in this dataset contained at least one region with AD previously marked by an expert radiologist. The results showed that the proposed method can reduce the false positives of the segmentation step of the CAD scheme from 43.4 false positives (FP) per image to 34.5 FP per image, without increasing the number of false negatives.

Automatic detection of the nipple in mammograms is an important step in computerized systems that combine multiview information for accurate detection and diagnosis of breast cancer. Locating the nipple is a difficult task owing to variations in image quality, presence of noise, and distortion and displacement of the breast tissue due to compression. In this work, we propose a novel Hessian-based method to locate automatically the nipple in screen-film and full-field digital mammograms (FFDMs). The method includes detection of a plausible nipple/retroareolar area in a mammogram using geometrical constraints, analysis of the gradient vector field by mean and Gaussian curvature measurements, and local shape-based conditions. The proposed procedure was tested on 566 mammographic images consisting of 372 randomly selected scanned films from two public databases (mini-MIAS and DDSM), and 194 digital mammograms acquired with a GE Senographe 2000D FFDM system. A radiologist independently marked the centers of the nipples for evaluation of the results. The average error obtained was 6.7 mm (22 pixels) with reference to the center of the nipple as identified by the radiologist. Only two out of the 566 detected nipples (0.35 %) had an error larger than 50 mm. The method was also directly compared with two other techniques for the detection of the nipple. The results indicate that the proposed method outperforms other algorithms presented in the literature and can be used to identify accurately the nipple on various types of mammographic images.

In clinical practice, the constructive consultation among experts improves the reliability of the diagnosis and leads to the definition of the treatment plan for the patient. Aggregation of the different opinions collected by many experts can be performed at the level of patient information, abnormality delineation, or final assessment. In this study, we present a novel cooperative strategy that exploits the dynamic contribution of the classification models composing the ensemble to make the final class assignment. As a proof of concept, we applied the proposed approach to the assessment of malignant infiltration in 103 vertebral compression fractures in magnetic resonance images. The results obtained with repeated random subsampling and receiver operating characteristic analysis indicate that the cooperative system statistically improved ( $$p

At present, mammography is the most effective examination for an early diagnosis of breast cancer. Nevertheless, the detection of cancer signs in mammograms is a difficult procedure owing to the great number of non-pathological structures which are also present in the image. Recent statistics show that in current breast cancer screenings 10%-25% of the tumors are missed by the radiologists. For this reason, a lot of research is currently being done to develop systems for Computer Aided Detection (CADe). Probably, some causes of the false-negative screening examinations are that tumoral masses have varying dimension and irregular shape, their borders are often ill-defined and their contrast is very low, thus making difficult the discrimination from parenchymal structures. Therefore, in a CADe system a preliminary segmentation procedure has to be implemented in order to separate the mass from the background tissue. In this way, various characteristics of the segmented mass can be evaluated and used in a classification step to discriminate benign and malignant cases. In this paper, we describe an effective algorithm for massive lesions segmentation based on a region-growing technique and we provide full details the performance evaluation procedure used in this specific context.

Summary: A major challenge in cancer research is the complexity of the tumor microenvironment, which includes the host immunological setting. Inspired by the emerging technology of organ-on-chip, we achieved 3D co-cultures in microfluidic devices (integrating four cell populations: cancer, immune, endothelial, and fibroblasts) to reconstitute ex vivo a human tumor ecosystem (HER2+ breast cancer). We visualized and quantified the complex dynamics of this tumor-on-chip, in the absence or in the presence of the drug trastuzumab (Herceptin), a targeted antibody therapy directed against the HER2 receptor. We uncovered the capacity of the drug trastuzumab to specifically promote long cancer-immune interactions (>50 min), recapitulating an anti-tumoral ADCC (antibody-dependent cell-mediated cytotoxicity) immune response. Cancer-associated fibroblasts (CAFs) antagonized the effects of trastuzumab. These observations constitute a proof of concept that tumors-on-chip are powerful platforms to study ex vivo immunocompetent tumor microenvironments, to characterize ecosystem-level drug responses, and to dissect the roles of stromal components. : Inspired by the emerging technology of tumor-on-chip, Nguyen et al. reconstituted ex vivo a human tumor microenvironment (HER2+ breast cancer), characterized the ecosystem-level responses to the drug trastuzumab (Herceptin), and dissected the roles of stromal components (immune cells and fibroblasts), demonstrating the power of immunocompetent tumors-on-chip for preclinical drug studies. Keywords: tumor microenvironment, organ-on-chip, tumor-on-chip, trastuzumab, HER2+ breast cancer, cancer-associated fibroblasts, live cell imaging, microfluidics, pre-clinical models, immunotherapy

In this paper, we perform the assessment of a CAD for the tumoral masses classification in mammograms by the uncertainty propagation through the system. Carrying on the work of the authors concerning the metrological characterization of the developed CAD, we validate the features extraction, features selection, and classification steps in this paper. In particular, suitable metrics such as the Receiving Operating Curve (ROC) and the Area Under ROC (AUC) are widely used in order to provide a quantitative evaluation of the performance. Finally, we implement a Monte Carlo simulation in order to provide the confidence interval for some coverage probabilities for all involved parameters. The procedure is tested on mammographic images containing both malignant and benign breast masses.

Breast cancer is the main cause of cancer deaths among women in the world. About one million new cases appear every year, and about 25% of them lead to the death of the patient. The best solution is the early detection of suspicious tumoral signs through an effective mammographic screening program. Unfortunately, this kind of images is very difficult to interpret by the radiologists because of its very low contrast, so proper image-processing procedures could help them to achieve better diagnoses. This paper improves and assesses an algorithm, already proposed by the authors, that suggests to doctors the suspicious regions that could contain tumoral masses. The procedure succeeds also in the case of very low contrast because it depends only on the orientation of the gradient vectors in the image but not on their amplitude.

Mammography is the most effective method for the early detection of breast diseases. However, the typical diagnostic signs such as microcalcifications and masses are difficult to detect because mammograms are low-contrast and noisy images. In this paper, a novel algorithm for image denoising and enhancement based on dyadic wavelet processing is proposed. The denoising phase is based on a local iterative noise variance estimation. Moreover, in the case of microcalcifications, we propose an adaptive tuning of enhancement degree at different wavelet scales, whereas in the case of mass detection, we developed a new segmentation method combining dyadic wavelet information with mathematical morphology. The innovative approach consists of using the same algorithmic core for processing images to detect both microcalcifications and masses. The proposed algorithm has been tested on a large number of clinical images, comparing the results with those obtained by several other algorithms proposed in the literature through both analytical indexes and the opinions of radiologists. Through preliminary tests, the method seems to meaningfully improve the diagnosis in the early breast cancer detection with respect to other approaches.

The interpretation of diagnostic images is often conditioned by the specific properties of the instrument that generated the image. This makes particularly complicated to develop universal recognition algorithms that can facilitate the diagnosis in case of massive population screenings. Mammography is a typical example where such an algorithm is required. Although the technological advances in medical imaging increases the accuracy of interpretation of images, the improved resolution may not facilitate the identification of breast cancer at a very early stage due to the many confounding factors related, for instance, to differences in instrument settings or breast positioning by the operator. Being impossible an exact standardization, the problem of reducing the effects of different instruments and operators can be faced with a proper algorithm that can extract from each image the relevant information in an unsupervised manner, thus limiting the influence of instrumental and positioning issues. For this scope, in this paper we investigated the properties of a classifier based on an ensemble of Adaptive Artificial Immune Networks (A2INET) applied to original mammography image indicators aimed at diagnosing bilateral asymmetry that is known to be correlated with increased breast cancer risk. Classification models were trained using a set of descriptors measuring the degree of similarity of paired regions of the left and right breasts. Noteworthy, the ensemble of A2INET models achieved very high classification rates even when training and testing were made on two completely independent and heterogeneous datasets. The obtained results are promising (maximum accuracy level of 0.90, sensitivity of 0.93 and specificity of 0.87) and they prefigure to apply automatic diagnostic tools in clinical practice exploiting a network of different instrument databases.

Early stage asymmetric signs in breast that can be captured by the screening-digital mammography can be used for a precocious diagnosis of breast cancer. Conventional mammography screening fails to detect subtle anomalies, so computer-aided methods are studied in order to improve the accuracy of image analysis. To classify the images into asymmetric and normal cases, in this paper we investigated the performance of an Adaptive Artificial Immune System (A2INET) classifier. To test the efficiency of the algorithm, two public datasets have been considered: 32 pairs of mammographic images including MLO projection retrieved from Digital Database for Screening mammographic (DDSM) and 30 ones from Mammographic Image Analysis Society (mini-MIAS) databases. Results show that A2INET yields best results with respect to the other more conventional classifiers.

We present a novel method to detect asymmetry in mammograms based upon bilateral analysis of the spatial distribution of density within paired mammographic strips. Various differential measures of spatial correlation of gray-scale values were computed with reference to the position of the nipple for a set of 128 pairs of mammograms from the Digital Database for Screening Mammography (DDSM). Features were selected by stepwise logistic regression and the leave-one-patient-out method was used for cross-validation of results. An area under the receiver operating characteristic curve of 0.87 (SE = 0.08) was achieved by using an artificial neural network classifier with radial basis functions.

We present new feature descriptors specifically designed to quantify angular nonstationarity and angular dependence of pixel values in sectors of mammographic lesions. A key novelty of this work is that the proposed measures characterize the texture of masses without relying on accurate determination of their contours. An artificial neural network based on radial basis functions was used to predict the diagnosis of 120 benign masses and 26 malignant tumors in a database of full-field digital mammograms. Features were selected using stepwise logistic regression and the leave-one-patient-out method was used for cross-validation of results. An area under the receiver operating characteristic curve of 0.9890 ± 0.0114 was obtained using randomly selected centroids and an expected size of the masses. Results indicate that the use of the proposed contour-independent features can be an effective approach for computer-aided classification of mammographic lesions.

Automatic tools for detection and identification of lung and lesion from high-resolution CT (HRCT) are becoming increasingly important both for diagnosis and for delivering high-precision radiation therapy. However, development of robust and interpretable classifiers still presents a challenge especially in case of non-small cell lung carcinoma (NSCLC) patients. In this paper, we have attempted to devise such a classifier by extracting fuzzy rules from texture segmented regions from HRCT images of NSCLC patients. A fuzzy inference system (FIS) has been constructed starting from a feature extraction procedure applied on overlapping regions from the same organs and deriving simple if–then rules so that more linguistically interpretable decisions can be implemented. The proposed method has been tested on 138 regions extracted from CT scan images acquired from patients with lung cancer. Assuming two classes of tissues C1 (healthy tissues) and C2 (lesion) as negative and positive, respectively; preliminary results report an AUC = 0.98 for lesions and AUC = 0.93 for healthy tissue, with an optimal operating condition related to sensitivity = 0.96, and specificity = 0.98 for lesions and sensitivity 0.99, and specificity = 0.94 for healthy tissue. Finally, the following results have been obtained: false-negative rate (FNR) = 6 % (C1), FNR = 2 % (C2), false-positive rate (FPR) = 4 % (C1), FPR = 3 % (C2), true-positive rate (TPR) = 94 %, (C1) and TPR = 98 % (C2).

Based on recent statistics from the International Agency for Research on Cancer (Ferlay et al., 2010), breast cancer account for 10.9% of all cancers diagnosed and ranks as the fifth cause of cancer death in the world. Although incidence rates are increasing, mortality rates are stable, representing an improved survival rate. This improvement can be attributed to effective means for the early detection as well as to significant improvement in treatment options, exposure, etc. Mammography is, at present, the only viable method for detecting most of tumors early enough for effective treatment, without unnecessary biopsies, or other invasive procedures. Therefore, screening mammography in women aged 40 to 70 years is currently the effective strategy to reduce breast cancer mortality. Early detection of invasive breast cancers is associated with better prognosis than waiting for women to become symptomatic. However, detecting the early signs of breast cancer is challenging because the cancerous structures have many features in common with normal breast tissue. Moreover, the accuracy of interpretation of screening mammograms is affected by several factors, such as image quality, the radiologist’s level of expertise, and the high volume of cases. Recent statistics show that in current breast cancer screenings 10% − 25% of the tumors are missed by the radiologists (Burrell et al., 2001; Cheng, 2003; Nishikawa, 2007). Missed cancers are due to many reasons: low disease prevalence, breast structure complexity, finding subtleties, and radiologist fatigue. To accomplish with these difficulties, different methods have been analyzed: first of all, double reading, which provides either double perception or double interpretation of lesions. It has been demonstrated that a single radiologist is more accurate when reading mammograms methodically than quickly and that two observers achieve an improvement in detection rate of 5% − 15% (Mazzarella & Bazzocchi, 2007). Obviously, this procedure is too expensive, complex, and time consuming especially in screening programs where a huge number of mammographic images have to be read. The development of computerized systems as second readers represents an alternative. Researchers have been developing algorithms to detect mammographic abnormalities for more than 30 years with the aim of either automating mammographic interpretation or providing a tool that could enhance human film-reading accuracy. Computer-aided detection (CADe) and diagnosis (CADx) systems are widely used in mammography, where signs of breast cancer are often very subtle. Both systems involve the use of computer algorithms to detect patterns in images associated with signs of disease and to assign them a malignancy index. This result 15

In this work we propose a joint two-side masking procedure for automatic analysis of mammographic images. The primary objective is the improvement of computerized systems capability in revealing additional findings, as the asymmetrical changes of the breast parenchyma. The method allows the proper comparison of the left and right breast by progressive selection of paired small areas on the mammogram, primarily the so-called “forbidden areas”, zones that need special attention in mammographic interpretation. The masking of specific areas of the mammogram requires the identification of two anatomical structures of the breast: the pectoral muscle and the nipple used, together with the breast skin line, to find paired matching points on the images for comparison. With this purpose, specific algorithms have been developed. In particular, a new method for nipple extraction will be presented and validated by expert radiologists, by the use of a proprietary program developed by the authors. Finally, an application example of the automatic Tabar masking procedure will be shown, in order to point out the potential of this method in detection of suspicious areas in mammograms.

Patients in the Intensive Care Unit (ICU) require huge resources because of the dysfunction of several of their vital organs. The heterogeneity and complexity of the ICU patients have generated interest in systems able to measure severity of illness as a method of predicting outcome, compare quality-of-care, analyze the costs of treating critically ill patients, evaluate stratification of clinical trials. The present work aims at developing a new scoring system to assess the severity of illness of patients admitted to general ICUs, by an observational, prospective, cohort study. The model will be developed in the period of study considering a population of the general ICU at the University Hospital Polyclinic of Tor Vergata so that the data obtained will be usable for Italian health care resource management and ICU performance evaluation within the hospital, the region, and nationwide. The proposed scoring system will revise the presently available systems, in the attempt to overcome their major drawbacks in the definition of the input variables, in the mathematical model employed to process them and the considered outcome. An important and qualifying point of this work lies in the attention paid to quantify and process the different sources of uncertainty, originated by the approximations introduced in the considered model and in the collection stage of the predictor variables.

In this paper, we propose a novel approach for the automatic breast boundary segmentation using spatial fuzzy c-means clustering and active contours models. We will evaluate the performance of the approach on screen film mammographic images digitized by specific scanner devices and full-field digital mammographic images at different spatial and pixel resolutions. Expert radiologists have supplied the reference boundary for the massive lesions along with the biopsy proven pathology assessment. A performance assessment procedure will be developed considering metrics such as precision, recall, F-measure, and accuracy of the segmentation results. A Montecarlo simulation will be also implemented to evaluate the sensitivity of the boundary extracted on the initial settings and on the image noise.

Breast cancer is the second most common cancer overall and the leading cause of cancer deaths in women. Mammography is, at present, the only viable method for detecting most of tumors early enough for effective treatment. The secret of setting up the accurate diagnosis is to detect and understand the most subtle signs of breast lesions. Analysis of asymmetry between the left and right mammograms can provide clues about the presence of early signs of tumors. In this work we present an automated procedure for bilateral asymmetry detection composed of the following steps: (1) mammography density analysis and fibro-glandular disc detection through adaptive clustering techniques, (2) analysis and implementation of bilateral asymmetries detection algorithms based on Gabor filters analysis, (3) use of a linear Bayes classifier with the leave-one-out method to asses the asymmetry degree of the two breasts, (4) metrological evaluation of the whole system through random and systematic measurement uncertainty contributions modeling.

Microcalcifications clusters are subtle signs of breast cancer and are very difficult to diagnose in the mammographic images because of their small size and low contrast with respect to the normal breast tissue. In this paper, we propose an automatic procedure for the identification of microcalcifications clusters to aid the radiologist in the detection of this kind of breast cancer signs.

Breast cancer is the most common cancer in women. Fortunately, in the last years, the percentage of recovery has become higher and higher. The reasons are multiple: prevention, better surgery techniques, better diagnoses. In order to improve diagnostic performance, many Computer Aided Diagnosis systems have been developed in the latest years, which help the radiologists to detect and classify lump-masses. These systems, however, do not yet consider the uncertainty associated to the measurements (the mammography), even if, from a metrological point of view, they should. In this paper, measurement uncertainty is considered through a fuzzy inference system for the classification of microcalcifications in digital mammography.

Radiologists that analyze screening mammographic images miss the 10–20% of the diagnosis since this kind of images are very difficult to interpret. In this paper, we present the first step of a CADx (Computer Aided Diagnosis) system that, from the original mammogram, extracts suspicious regions on which the radiologists have to focus their attention. The procedure often successes also in case of very low contrast, because it depends only on the orientation of the gradient vectors in the image but not on their amplitude.

The paper presents a first implementation of a novel tool to assist radiologists in analyzing screening mammographic images. The software named Assisted Breast Cancer Diagnosis Environment (ABCDE) is able to acquire DICOM images (also in presence of Grayscale Softcopy Presentation State). It is designed to assist the doctor, as a second reader, during the various phases of the diagnosis. The program uses some public libraries and implements many new algorithms developed in the last years by the research group.

In this paper, we consider uncertainty handling and propagation by means of random fuzzy variables (RFVs) through a computer-aided-diagnosis (CADx) system for the early diagnosis of breast cancer. In particular, the denoising and the contrast enhancement of microcalcifications is specifically addressed, providing a novel methodology for separating the foreground and the background in the image to selectively process them. The whole system is then assessed by metrological aspects. In this context, we assume that the uncertainty associated to each pixel of the image has both a random and a non-negligible systematic contribution. Consequently, a preliminary noise variance estimation is performed on the original image, and then, using suitable operators working on RFVs, the uncertainty propagation is evaluated through the whole system. Finally, we compare our results with those obtained by a Monte Carlo method.

Breast cancer is the most common cause of death among women and the most effective method for its diagnosis is mammography. However, this kind of analysis is very difficult to interpret and for this reason radiologists miss 20-30% of tumors. We propose a module for the segmentation of masses that can be implemented in a complete CADx (Computer Aided Diagnosis) system. In particular, we implement a new version of the region growing algorithm specific for this kind of images and for the constraints on computation time of this application.