Intelligent Diagnosis of Thyroid Ultrasound Imaging Using an Ensemble of Deep Learning Methods.

Background and Objectives: At present, thyroid disorders have a great incidence in the worldwide population, so the development of alternative methods for improving the diagnosis process is necessary. Materials and Methods: For this purpose, we developed an ensemble method that fused two deep learning models, one based on convolutional neural network and the other based on transfer learning. For the first model, called 5-CNN, we developed an efficient end-to-end trained model with five convolutional layers, while for the second model, the pre-trained VGG-19 architecture was repurposed, optimized and trained. We trained and validated our models using a dataset of ultrasound images consisting of four types of thyroidal images: autoimmune, nodular, micro-nodular, and normal. Results: Excellent results were obtained by the ensemble CNN-VGG method, which outperformed the 5-CNN and VGG-19 models: 97.35% for the overall test accuracy with an overall specificity of 98.43%, sensitivity of 95.75%, positive and negative predictive value of 95.41%, and 98.05%. The micro average areas under each receiver operating characteristic curves was 0.96. The results were also validated by two physicians: an endocrinologist and a pediatrician. Conclusions: We proposed a new deep learning study for classifying ultrasound thyroidal images to assist physicians in the diagnosis process.

Feasibility of a Portable Abdominal Insufflation Device for Controlling Intraperitoneal Bleeding After Abdominal Blunt Trauma.

Uncontrolled bleeding contributes to 30% to 40% of trauma-related deaths and is the leading cause of potentially preventable deaths. Currently, there is no effective method available to first responders for temporary control of noncompressible intraabdominal bleeding while patients are transported to the hospital. Our previous studies demonstrated that abdominal insufflation provides effective temporary bleeding control. The study aims to prove the feasibility (insufflation to a target pressure) and safety (cardiovascular and respiratory effects) of a novel portable abdominal insufflation device (PAID) designed to control the intraperitoneal bleeding caused by abdominal trauma. The PAID prototype is based on a patented design and manufactured via additive manufacturing. PAID contains a 16-g CO2 cartridge and an electronic pressure transducer. PAID was tested on a bench top and a swine animal model. For the animal model study, the intraperitoneal pressure as well as cardiorespiratory parameters (hearth rate, SpO2 [peripheral capillary oxygen saturation], and blood pressure) were continuously monitored during the insufflation procedure. The prototype functioned according to specifications on both bench top and animal models. CO2 insufflation of the peritoneal cavity was delivered up the target 20 mm Hg and maintained for 30 minutes from 1 or 2 cartridges in the swine model. No intraoperative incidents were registered, and all the recorded physiological parameters were within normal limits. The PAID prototype is a feasible, easy to use device that provides quick, controlled, and safe insufflation of the peritoneal cavity. Future studies will focus on testing the next-generation, semiautomatic PAID prototype in a severe intraabdominal injury model.

Experimental and theoretical model of microvascular network remodeling and blood flow redistribution following minimally invasive microvessel laser ablation.

Overly dense microvascular networks are treated by selective reduction of vascular elements. Inappropriate manipulation of microvessels could result in loss of host tissue function or a worsening of the clinical problem. Here, experimental, and computational models were developed to induce blood flow changes via selective artery and vein laser ablation and study the compensatory collateral flow redistribution and vessel diameter remodeling. The microvasculature was imaged non-invasively by bright-field and multi-photon laser microscopy, and optical coherence tomography pre-ablation and up to 30 days post-ablation. A theoretical model of network remodeling was developed to compute blood flow and intravascular pressure and identify vessels most susceptible to changes in flow direction. The skin microvascular remodeling patterns were consistent among the five specimens studied. Significant remodeling occurred at various time points, beginning as early as days 1-3 and continuing beyond day 20. The remodeling patterns included collateral development, venous and arterial reopening, and both outward and inward remodeling, with variations in the time frames for each mouse. In a representative specimen, immediately post-ablation, the average artery and vein diameters increased by 14% and 23%, respectively. At day 20 post-ablation, the maximum increases in arterial and venous diameters were 2.5× and 3.3×, respectively. By day 30, the average artery diameter remained 11% increased whereas the vein diameters returned to near pre-ablation values. Some arteries regenerated across the ablation sites via endothelial cell migration, while veins either reconnected or rerouted flow around the ablation site, likely depending on local pressure driving forces. In the intact network, the theoretical model predicts that the vessels that act as collaterals after flow disruption are those most sensitive to distant changes in pressure. The model results correlate with the post-ablation microvascular remodeling patterns.

Experimental and Theoretical Model of Single Vessel Minimally Invasive Micro-Laser Ablation: Inducing Microvascular Network Remodeling and Blood Flow Redistribution Without Compromising Host Tissue Function.

Overly dense microvascular networks are treated by selective reduction of vascular elements. Inappropriate manipulation of microvessels could result in loss of host tissue function or a worsening of the clinical problem. Here, experimental, and computational models were developed to induce blood flow changes via selective artery and vein laser ablation and study the compensatory collateral flow redistribution and vessel diameter remodeling. The microvasculature was imaged non-invasively by bright-field and multi-photon laser microscopy, and Optical Coherence Tomography pre-ablation and up to 30 days post-ablation. A theoretical model of network remodeling was developed to compute blood flow and intravascular pressure and identify vessels most susceptible to changes in flow direction. The skin microvascular remodeling patterns were consistent among the five specimens studied. Significant remodeling occurred at various time points, beginning as early as days 1-3 and continuing beyond day 20. The remodeling patterns included collateral development, venous and arterial reopening, and both outward and inward remodeling, with variations in the time frames for each mouse. In a representative specimen, immediately post-ablation, the average artery and vein diameters increased by 14% and 23%, respectively. At day 20 post-ablation, the maximum increases in arterial and venous diameters were 2.5x and 3.3x, respectively. By day 30, the average artery diameter remained 11% increased whereas the vein diameters returned to near pre-ablation values. Some arteries regenerated across the ablation sites via endothelial cell migration, while veins either reconnected or rerouted flow around the ablation site, likely depending on local pressure driving forces. In the intact network, the theoretical model predicts that the vessels that act as collaterals after flow disruption are those most sensitive to distant changes in pressure. The model results match the post-ablation microvascular remodeling patterns.

Structural remodeling of the mouse gracilis artery: coordinated changes in diameter and medial area maintain circumferential stress.

OBJECTIVE: Vascular networks respond to chronic alterations in blood supply by structural remodeling. Previously, we showed that blood flow changes in the mouse GA lead to transient diameter increases, which can generate large increases in circumferential wall stress. Here, we examine the associated changes in the medial area of the arterial wall and the effects on circumferential wall stress. METHODS: To induce blood flow changes, one of the two feeding vessels to the GA was surgically removed. At 7-56 days after blood flow interruption, the vasculature was perfused with India ink for morphological measurements, and processed for immuno-cytochemistry to mark the medial cross-section area. Theoretical simulations of hemodynamics were used to analyze the data. RESULTS: During adaptive increases in vessel diameter, increases in medial area were observed, most strongly in the middle region of the artery. Simulations showed that this increase in medial area limits the increase in estimated circumferential stress during vascular adaptation to less than 50%, in contrast to an increase of up to 250% if the medial area had remained unchanged. CONCLUSIONS: During vascular adaptation, increases in circumferential stress are limited by growth of the media coordinated with diameter changes.

Feasibility of a lung airway navigation system using fiber-Bragg shape sensing and artificial intelligence for early diagnosis of lung cancer.

Currently early diagnosis of malignant lesions at the periphery of lung parenchyma requires guidance of the biopsy needle catheter from the bronchoscope into the smaller peripheral airways via harmful X-ray radiation. Previously, we developed an image-guided system, iMTECH which uses electromagnetic tracking and although it increases the precision of biopsy collection and minimizes the use of harmful X-ray radiation during the interventional procedures, it only traces the tip of the biopsy catheter leaving the remaining catheter untraceable in real time and therefore increasing image registration error. To address this issue, we developed a shape sensing guidance system containing a fiber-Bragg grating (FBG) catheter and an artificial intelligence (AI) software, AIrShape to track and guide the entire biopsy instrument inside the lung airways, without radiation or electromagnetic navigation. We used a FBG fiber with one central and three peripheral cores positioned at 120° from each other, an array of 25 draw tower gratings with 1cm/3nm spacing, 2 mm grating length, Ormocer-T coating, and a total outer diameter of 0.2 mm. The FBG fiber was placed in the working channel of a custom made three-lumen catheter with a tip bending mechanism (FBG catheter). The AIrShape software determines the position of the FBG catheter by superimposing its position to the lung airway center lines using an AI algorithm. The feasibility of the FBG system was tested in an anatomically accurate lung airway model and validated visually and with the iMTECH platform. The results prove a viable shape-sensing hardware and software navigation solution for flexible medical instruments to reach the peripheral airways. During future studies, the feasibility of FBG catheter will be tested in pre-clinical animal models.

Image Fusion Involving Real-Time Transabdominal or Endoscopic Ultrasound for Gastrointestinal Malignancies: Review of Current and Future Applications.

Image fusion of CT, MRI, and PET with endoscopic ultrasound and transabdominal ultrasound can be promising for GI malignancies as it has the potential to allow for a more precise lesion characterization with higher accuracy in tumor detection, staging, and interventional/image guidance. We conducted a literature review to identify the current possibilities of real-time image fusion involving US with a focus on clinical applications in the management of GI malignancies. Liver applications have been the most extensively investigated, either in experimental or commercially available systems. Real-time US fusion imaging of the liver is gaining more acceptance as it enables further diagnosis and interventional therapy of focal liver lesions that are difficult to visualize using conventional B-mode ultrasound. Clinical studies on EUS guided image fusion, to date, are limited. EUS-CT image fusion allowed for easier navigation and profiling of the target tumor and/or surrounding anatomical structure. Image fusion techniques encompassing multiple imaging modalities appear to be feasible and have been observed to increase visualization accuracy during interventional and diagnostic applications.

Deep Learning Algorithm for the Confirmation of Mucosal Healing in Crohn's Disease, Based on Confocal Laser Endomicroscopy Images.

BACKGROUND AND AIMS: Mucosal healing (MH) is associated with a stable course of Crohn's disease (CD) which can be assessed by confocal laser endomicroscopy (CLE). To minimize the operator's errors and automate assessment of CLE images, we used a deep learning (DL) model for image analysis. We hypothesized that DL combined with convolutional neural networks (CNNs) and long short-term memory (LSTM) can distinguish between normal and inflamed colonic mucosa from CLE images. METHODS: The study included 54 patients, 32 with known active CD, and 22 control patients (18 CD patients with MH and four normal mucosa patients with no history of inflammatory bowel diseases). We designed and trained a deep convolutional neural network to detect active CD using 6,205 endomicroscopy images classified as active CD inflammation (3,672 images) and control mucosal healing or no inflammation (2,533 images). CLE imaging was performed on four colorectal areas and the terminal ileum. Gold standard was represented by the histopathological evaluation. The dataset was randomly split in two distinct training and testing datasets: 80% data from each patient were used for training and the remaining 20% for testing. The training dataset consists of 2,892 images with inflammation and 2,189 control images. The testing dataset consists of 780 images with inflammation and 344 control images of the colon. We used a CNN-LSTM model with four convolution layers and one LSTM layer for automatic detection of MH and CD diagnosis from CLE images. RESULTS: CLE investigation reveals normal colonic mucosa with round crypts and inflamed mucosa with irregular crypts and tortuous and dilated blood vessels. Our method obtained a 95.3% test accuracy with a specificity of 92.78% and a sensitivity of 94.6%, with an area under each receiver operating characteristic curves of 0.98. CONCLUSIONS: Using machine learning algorithms on CLE images can successfully differentiate between inflammation and normal ileocolonic mucosa and can be used as a computer aided diagnosis for CD. Future clinical studies with a larger patient spectrum will validate our results and improve the CNN-SSTM model.

Transfer learning with pre-trained deep convolutional neural networks for the automatic assessment of liver steatosis in ultrasound images.

AIM: In this paper we proposed different architectures of convolutional neural network (CNN) to classify fatty liver disease in images using only pixels and diagnosis labels as input. We trained and validated our models using a dataset of 629 images consisting of 2 types of liver images, normal and liver steatosis. MATERIAL AND METHODS: We assessed two pre-trained models of convolutional neural networks, Inception-v3 and VGG-16 using fine-tuning. Both models were pre-trained on ImageNet dataset to extract features from B-mode ultrasound liver images. The results obtained through these methods were compared for selecting the predictive model with the best performance metrics. We trained the two models using a dataset of 262 images of liver steatosis and 234 images of normal liver. We assessed the models using a dataset of 70 liver steatosis im-ages and 63 normal liver images. RESULTS: The proposed model that used Inception v3 obtained a 93.23% test accuracy with a sensitivity of 89.9%% and a precision of 96.6%, and areas under each receiver operating characteristic curves (ROC AUC) of 0.93. The other proposed model that used VGG-16, obtained a 90.77% test accuracy with a sensitivity of 88.9% and a precision of 92.85%, and areas under each receiver operating characteristic curves (ROC AUC) of 0.91. CONCLUSION: The deep learning algorithms that we proposed to detect steatosis and classify the images in normal and fatty liver images, yields an excellent test performance of over 90%. However, future larger studies are required in order to establish how these algorithms can be implemented in a clinical setting.

Assessment of Deep Learning Methods for Differentiating Autoimmune Disorders in Ultrasound Images.

At present, deep learning becomes an important tool in medical image analysis, with good performance in diagnosing, pattern detection, and segmentation. Ultrasound imaging offers an easy and rapid method to detect and diagnose thyroid disorders. With the help of a computer-aided diagnosis (CAD) system based on deep learning, we have the possibility of real-time and non-invasive diagnosing of thyroidal US images. This paper proposed a study based on deep learning with transfer learning for differentiating the thyroidal ultrasound images using image pixels and diagnosis labels as inputs. We trained, assessed, and compared two pre-trained models (VGG-19 and Inception v3) using a dataset of ultrasound images consisting of 2 types of thyroid ultrasound images: autoimmune and normal. The training dataset consisted of 615 thyroid ultrasound images, from which 415 images were diagnosed as autoimmune, and 200 images as normal. The models were assessed using a dataset of 120 images, from which 80 images were diagnosed as autoimmune, and 40 images diagnosed as normal. The two deep learning models obtained very good results, as follows: the pre-trained VGG-19 model obtained 98.60% for the overall test accuracy with an overall specificity of 98.94% and overall sensitivity of 97.97%, while the Inception v3 model obtained 96.4% for the overall test accuracy with an overall specificity of 95.58% and overall sensitivity of 95.58.

Real-time computer-aided diagnosis of focal pancreatic masses from endoscopic ultrasound imaging based on a hybrid convolutional and long short-term memory neural network model.

Differential diagnosis of focal pancreatic masses is based on endoscopic ultrasound (EUS) guided fine needle aspiration biopsy (EUS-FNA/FNB). Several imaging techniques (i.e. gray-scale, color Doppler, contrast-enhancement and elastography) are used for differential diagnosis. However, diagnosis remains highly operator dependent. To address this problem, machine learning algorithms (MLA) can generate an automatic computer-aided diagnosis (CAD) by analyzing a large number of clinical images in real-time. We aimed to develop a MLA to characterize focal pancreatic masses during the EUS procedure. The study included 65 patients with focal pancreatic masses, with 20 EUS images selected from each patient (grayscale, color Doppler, arterial and venous phase contrast-enhancement and elastography). Images were classified based on cytopathology exam as: chronic pseudotumoral pancreatitis (CPP), neuroendocrine tumor (PNET) and ductal adenocarcinoma (PDAC). The MLA is based on a deep learning method which combines convolutional (CNN) and long short-term memory (LSTM) neural networks. 2688 images were used for training and 672 images for testing the deep learning models. The CNN was developed to identify the discriminative features of images, while a LSTM neural network was used to extract the dependencies between images. The model predicted the clinical diagnosis with an area under curve index of 0.98 and an overall accuracy of 98.26%. The negative (NPV) and positive (PPV) predictive values and the corresponding 95% confidential intervals (CI) are 96.7%, [94.5, 98.9] and 98.1%, [96.81, 99.4] for PDAC, 96.5%, [94.1, 98.8], and 99.7%, [99.3, 100] for CPP, and 98.9%, [97.5, 100] and 98.3%, [97.1, 99.4] for PNET. Following further validation on a independent test cohort, this method could become an efficient CAD tool to differentiate focal pancreatic masses in real-time.

Finite Element Analysis of a Novel Aortic Valve Stent.

Worldwide, one of the leading causes of death for patients with cardiovascular disease is aortic valve failure or insufficiency as a result of calcification and cardiovascular disease. The surgical treatment consists of repair or total replacement of the aortic valve. Artificial aortic valve implantation via a percutaneous or endovascular procedure is the minimally invasive alternative to open chest surgery, and the only option for high-risk or older patients. Due to the complex anatomical location between the left ventricle and the aorta, there are still engineering design optimization challenges which influence the long-term durability of the valve. In this study we developed a computer model and performed a numerical analysis of an original self-expanding stent for transcatheter aortic valve in order to optimize its design and materials. The study demonstrates the current valve design could be a good alternative to the existing commercially available valve devices.

Preoperative Computer-Assisted Laparoscopy Planning for the Minimally Invasive Surgical Repair of Hiatal Hernia.

Minimal invasive surgical procedures such as laparoscopy are preferred over open surgery due to faster postoperative recovery, less trauma and inflammatory response, and less scarring. Laparoscopic repairs of hiatal hernias require pre-procedure planning to ensure appropriate exposure and positioning of the surgical ports for triangulation, ergonomics, instrument length and operational angles to avoid the fulcrum effect of the long and rigid instruments. We developed a novel surgical planning and navigation software, iMTECH to determine the optimal location of the skin incision and surgical instrument placement depth and angles during laparoscopic surgery. We tested the software on five cases of human hiatal hernia to assess the feasibility of the stereotactic reconstruction of anatomy and surgical planning. A whole-body CT investigation was performed for each patient, and abdominal 3D virtual models were reconstructed from the CT scans. The optical trocar access point was placed on the xipho-umbilical line. The distance on the skin between the insertion point of the optical trocar and the xiphoid process was 159.6, 155.7, 143.1, 158.3, and 149.1 mm, respectively, at a 40° elevation angle. Following the pre-procedure planning, all patients underwent successful surgical laparoscopic procedures. The user feedback was that planning software significantly improved the ergonomics, was easy to use, and particularly useful in obese patients with large hiatal defects where the insertion points could not be placed in the traditional positions. Future studies will assess the benefits of the planning system over the conventional, empirical trocar positioning method in more patients with other surgical challenges.

Skin Diseases Classification Using Deep Leaning Methods.

Due to the high incidence of skin tumors, the development of computer aided-diagnosis methods will become a very powerful diagnosis tool for dermatologists. The skin diseases are initially diagnosed visually, through clinical screening and followed in some cases by dermoscopic analysis, biopsy and histopathological examination. Automatic classification of dermatoscopic images is a challenge due to fine-grained variations in lesions. The convolutional neural network (CNN), one of the most powerful deep learning techniques proved to be superior to traditional algorithms. These networks provide the flexibility of extracting discriminatory features from images that preserve the spatial structure and could be developed for region recognition and medical image classification. In this paper we proposed an architecture of CNN to classify skin lesions using only image pixels and diagnosis labels as inputs. We trained and validated the CNN model using a public dataset of 10015 images consisting of 7 types of skin lesions: actinic keratoses and intraepithelial carcinoma/Bowen disease (akiec), basal cell carcinoma (bcc), benign lesions of the keratosis type (solar lentigine/seborrheic keratoses and lichen-planus like keratosis, bkl), dermatofibroma (df), melanoma (mel), melanocytic nevi (nv) and vascular lesions (angiomas, angiokeratomas, pyogenic granulomas and hemorrhages, vasc).

Vascular beds maintain pancreatic tumour explants for ex vivo drug screening.

Our understanding of cancer progression or response to therapies would benefit from benchtop, tissue-level assays that preserve the biology and anatomy of human tumours ex vivo. We present a methodology for maintaining patient tumour samples ex vivo for the purpose of drug testing in a clinical setting. The harvested tumour biopsy, excised from mice or patients, is integrated into a support tissue that includes stroma and vasculature. This support tissue preserves tumour histoarchitecture and relevant expression profiles, and tumour tissues cultured using this system display different sensitivities to chemotherapeutics compared with tumour explants with no supporting tissue. The methodology is more rapid than patient-derived xenograft models, easy to implement, and amenable to high-throughput assays, making it an attractive tool for in vitro drug screening or for the guidance of patient-specific chemotherapies.

Assessing tumor angiogenesis in colorectal cancer by quantitative contrast-enhanced endoscopic ultrasound and molecular and immunohistochemical analysis.

BACKGROUND AND OBJECTIVES: Data on contrast-enhanced endoscopic ultrasound (CE-EUS) for colorectal cancer (CRC) evaluation are scarce. Therefore, we aimed to assess the vascular perfusion pattern in CRC by quantitative CE-EUS and compare it to immunohistochemical and genetic markers of angiogenesis. PATIENTS AND METHODS: We performed a retrospective analysis of CE-EUS examinations of 42 CRC patients, before any therapy. CE-EUS movies were processed using a dedicated software. Ten parameters were automatically generated from the time-intensity curve (TIC) analysis: peak enhancement (PE), rise time (RT), mean transit time, time to peak (TTP), wash-in area under the curve (WiAUC), wash-in rate (WiR), wash-in perfusion index (WiPI), wash-out AUC (WoAUC), and wash-in and wash-out AUC (WiWoAUC). The expression levels of the vascular endothelial growth factor receptor 1 (VEGFR1) and VEGFR2 genes were assessed from biopsy samples harvested during colonoscopy. Microvascular density and vascular area were calculated after CD31 and CD105 immunostaining. RESULTS: Forty-two CE-EUS video sequences were analyzed. We found positive correlations between the parameters PE, WiAUC, WiR, WiPI, WoAUC, WiWoAUC, and N staging (Spearman r = 0.437, r = 0.336, r = 0.462, r = 0.437, r = 0.358, and r = 0.378, respectively, P < 0.05), and also between RT and TTP and CD31 vascular area (r = 0.415, and r = 0.421, respectively, P < 0.05). VEGFR1 and VEGFR2 expression did not correlate with any of the TIC parameters. CONCLUSIONS: CE-EUS with TIC analysis enables minimally invasive assessment of CRC angiogenesis and may provide information regarding the lymph nodes invasion. However, further studies are needed for defining its role in the evaluation of CRC patients.

Self-assembly of vascularized tissue to support tumor explants in vitro.

Testing the efficacy of cancer drugs requires functional assays that recapitulate the cell populations, anatomy and biological responses of human tumors. Although current animal models and in vitro cell culture platforms are informative, they have significant shortcomings. Mouse models can reproduce tissue-level and systemic responses to tumor growth and treatments observed in humans, but xenografts from patients often do not grow, or require months to develop. On the other hand, current in vitro assays are useful for studying the molecular bases of tumorigenesis or drug activity, but often lack the appropriate in vivo cell heterogeneity and natural microenvironment. Therefore, there is a need for novel tools that allow rapid analysis of patient-derived tumors in a robust and representative microenvironment. We have developed methodology for maintaining harvested tumor tissue in vitro by placing them in a support bed with self-assembled stroma and vasculature. The harvested biopsy or tumor explant integrates with the stromal bed and vasculature, providing the correct extracellular matrix (collagen I, IV, fibronectin), associated stromal cells, and a lumenized vessel network. Our system provides a new tool that will allow ex vivo drug-screening and can be adapted for the guidance of patient-specific therapeutic strategies.

A novel fusion imaging system for endoscopic ultrasound.

BACKGROUND AND OBJECTIVE: Navigation of a flexible endoscopic ultrasound (EUS) probe inside the gastrointestinal (GI) tract is problematic due to the small window size and complex anatomy. The goal of the present study was to test the feasibility of a novel fusion imaging (FI) system which uses electromagnetic (EM) sensors to co-register the live EUS images with the pre-procedure computed tomography (CT) data with a novel navigation algorithm and catheter. METHODS: An experienced gastroenterologist and a novice EUS operator tested the FI system on a GI tract bench top model. Also, the experienced gastroenterologist performed a case series of 20 patients during routine EUS examinations. RESULTS: On the bench top model, the experienced and novice doctors reached the targets in 67 ± 18 s and 150 ± 24 s with a registration error of 6 ± 3 mm and 11 ± 4 mm, respectively. In the case series, the total procedure time was 24.6 ± 6.6 min, while the time to reach the clinical target was 8.7 ± 4.2 min. CONCLUSIONS: The FI system is feasible for clinical use, and can reduce the learning curve for EUS procedures and improve navigation and targeting in difficult anatomic locations.

Computer Aided Diagnosis for Confocal Laser Endomicroscopy in Advanced Colorectal Adenocarcinoma.

INTRODUCTION: Confocal laser endomicroscopy (CLE) is becoming a popular method for optical biopsy of digestive mucosa for both diagnostic and therapeutic procedures. Computer aided diagnosis of CLE images, using image processing and fractal analysis can be used to quantify the histological structures in the CLE generated images. The aim of this study is to develop an automatic diagnosis algorithm of colorectal cancer (CRC), based on fractal analysis and neural network modeling of the CLE-generated colon mucosa images. MATERIALS AND METHODS: We retrospectively analyzed a series of 1035 artifact-free endomicroscopy images, obtained during CLE examinations from normal mucosa (356 images) and tumor regions (679 images). The images were processed using a computer aided diagnosis (CAD) medical imaging system in order to obtain an automatic diagnosis. The CAD application includes image reading and processing functions, a module for fractal analysis, grey-level co-occurrence matrix (GLCM) computation module, and a feature identification module based on the Marching Squares and linear interpolation methods. A two-layer neural network was trained to automatically interpret the imaging data and diagnose the pathological samples based on the fractal dimension and the characteristic features of the biological tissues. RESULTS: Normal colon mucosa is characterized by regular polyhedral crypt structures whereas malignant colon mucosa is characterized by irregular and interrupted crypts, which can be diagnosed by CAD. For this purpose, seven geometric parameters were defined for each image: fractal dimension, lacunarity, contrast correlation, energy, homogeneity, and feature number. Of the seven parameters only contrast, homogeneity and feature number were significantly different between normal and cancer samples. Next, a two-layer feed forward neural network was used to train and automatically diagnose the malignant samples, based on the seven parameters tested. The neural network operations were cross-entropy with the results: training: 0.53, validation: 1.17, testing: 1.17, and percent error, resulting: training: 16.14, validation: 17.42, testing: 15.48. The diagnosis accuracy error was 15.5%. CONCLUSIONS: Computed aided diagnosis via fractal analysis of glandular structures can complement the traditional histological and minimally invasive imaging methods. A larger dataset from colorectal and other pathologies should be used to further validate the diagnostic power of the method.

Implantable tissue isolation chambers for analyzing tumor dynamics in vivo.

Recruitment of new blood vessels from the surrounding tissue is central to tumor progression and involves a fundamental transition of the normal, organized vasculature into a dense disarray of vessels that infiltrates the tumor. At present, studying the co-development of the tumor and recruited normal tissue is experimentally challenging because many of the important events occur rapidly and over short length scales in a dense three-dimensional space. To overcome these experimental limitations, we partially confined tumors within biocompatible and optically clear tissue isolation chambers (TICs) and implanted them in mice to create a system that is more amenable to microscopic analysis. Our goal was to integrate the tumor into a recruited host tissue - complete with vasculature - and demonstrate that the system recapitulates relevant features of the tumor microenvironment. We show that the TICs allow clear visualization of the cellular events associated with tumor growth and progression at the host-tumor interface including cell infiltration, matrix remodeling and angiogenesis. The tissue within the chamber is viable for more than a month, and the process is robust in both the skin and brain. Treatment with losartan, an angiotensin II receptor antagonist, decreased the collagen density and fiber length in the TIC, consistent with the known activity of this drug. We further show that collagen fibers display characteristic tumor signatures and play a central role in angiogenesis, guiding the migration of tethered endothelial sprouts. The methodology combines accessible methods of microfabrication with animal models and will enable more informative studies of the cellular mechanisms of tumor progression.