- Email: [email protected]
Su2077 Clinical Study and Analysis of Ex Vivo Photoacoustic Imaging in Endoscopic Mucosal Resection Tissues in Barrett's Esophagus
GraphPad Prism. RESULTS: T2 relaxation time in visceral fat evaluation revealed a nonstatistically significant difference between celiac and obese patients (p>0.05), whereas a statistically significant difference was found between celiac patients and control patients (p<0.001), as well as between obese patients and control patients (p<0.001). T2 relaxation time in subcutaneous fat evaluation revealed a statistically significant difference between celiac and obese patients (p<0.01), as well as between celiac and control patients (p<0.001). No significant difference was found between obese and control patients (p>0.05). In addition, the ratio between visceral and subcutaneous T2 signal was higher in obese patients than in celiac patients (p<0.01). CONCLUSIONS: MRI can be suggested as complementary reproducible tool for the quantification of inflammatory state in celiac and obese patients. Increased visceral inflammatory signals were similar between celiac and obese patients, whereas subcutaneous T2 signals were higher in celiac patients than in obese patients. These data are probably due to the systemic inflammatory nature of CD. Concerning obesity, it may be confirmed the pro-inflammatory role of visceral fat alone, in respect of inactive subcutaneous fat depot.
Clinical Study and Analysis of Ex Vivo Photoacoustic Imaging in Endoscopic Mucosal Resection Tissues in Barrett's Esophagus Liang Lim, Catherine Streutker, Norman E. Marcon, Maria Cirocco, Alexandra Lao, Vladimir Iakovlev, Ralph DaCosta, Brian Wilson Accurate endoscopic detection of dysplasia in patients with Barrett's esophagus (BE) remains a major unmet clinical need. Current diagnosis uses multiple biopsies under endoscopic image guidance, where up to 99% of the tissue remains unsampled, leading to significant risk of missing dysplasia. We have conducted an ex vivo clinical trial using photoacoustic imaging (PAI) in BE patients undergoing endoscopic mucosal resection (EMR) with known high-grade dysplasia, for the purpose of characterizing the esophageal microvascular pattern. The long-term goal is to perform in vivo endoscopic PAI for dysplasia detection and staging by taking advantage of PAI's ability to image through the mucosa into the submucosa. Thirteen (13) tissue samples from 8 patients were analyzed, spanning a range of pathological classifications, including columnar type mucosa (5), dysplasia (5) and intramucosal adenocarcinoma (3). EMR tissues were mounted immediately after resection with the luminal side up on a clear agar slab and covered with ultrasound gel. The PAI transducer (40 MHz center frequency) was placed along the short axis of the tissue. Acoustic image slices (ultrasound and photoacoustic at 680, 750, 824, 850 and 970 nm) were simultaneously acquired covering the full length of the sample, each image slice having a field of view of 14 mm (width) by 15 mm (depth). The tissues were then sliced along the acoustic image slice at approximately 2 mm spacing and fixed in formalin for histopathology with H&E staining. From the acoustic images, we created 3D reconstruction of the full ex vivo tissue volume and generated images of the relative hemoglobin concentrations (oxy-, deoxy- and total) and oxygen saturation. We co-registered the acoustic images and the corresponding histological images, as illustrated in Figures 1A-B. The photoacoustic signal distribution within the tissue appears to coincide with the distribution of blood (dark pink on the H&E images), the main optical absorber in EMR tissue. These initial PAI + ultrasound images have demonstrated the technical feasibility of this approach and point to the potential of PAI to reveal the microvascular pattern within EMR specimens. Analysis of total hemoglobin and the intrinsic acoustic signals (Figure 1C-F) are currently being assessed against histologically diagnosed H&E images for each histopathological classification. These results will be reported and the limitations of this type of photoacoustic mapping of BE will be discussed.
Su2079 Computer Vision Analysis and Content Sampling for Intestinal Organoid Assays Ian A. Williamson, Adam D. Gracz, Scott Magness, Nancy Allbritton 3D intestinal organoid culture allows for real time observation of stem cell driven tissue renewal but impedes repetitive observation of individual cells/organoids in 3D space. Recently, we employed microraft array devices (MRAs) to spatially separating cells in microwells addressed to allow for recall of specific cells/organoids in one of the 15,000 microwells. We optimized CellProfiler computer vision pipelines to reduce user burden by automatically quantifying content and growth data from images of each microwell . First we made pipelines to accurately count the stem and niche cells initially present in each microwell to investigate the effect of cell-signaling interactions on organoid formation. We observed that organoid formation was only significantly increased when stem and niche cells were in physical contact suggesting that secreted factors were not sufficient for niche support. Organoids cultured long-term have widely variable appearances but display two distinct early morphologies discerned by the thickness of their outer monolayer. We developed further pipelines to quantify organoid monolayer width and identify the central pseudolumen compartment. Semi-automated organoid retrieval and downstream gene expression analysis showed significant differences between the two morphologies. Our MRA computer vision technologies will provide a basis for developing fully automated intestinal organoid based assays relating organoid growth/morphology to cell-signaling events.
Su2080 Limited Nesting Stress Alters Maternal Behavior and In Vivo Intestinal Permeability in a Sex-Dependent Manner in Wistar Rat Pups Nabila Moussaoui, Muriel H. Larauche, Mandy Biraud, Jenny Molet, Mulugeta Million, Emeran A. Mayer, Yvette Tache Background: In human, early-life adverse events predispose to psychiatric illness later in life and to functional gastrointestinal disorders such as irritable bowel syndrome. Experimental model of limited nesting stress (LNS) postnatally has been developed to recapitulate elements of human postnatal conditions where the mother is present but chronically provides erratic or unpredictable maternal care in an impoverished environment. Aims: To characterize the alterations of maternal behaviors induced by LNS in Wistar rats and to assess the impact in the offspring on components of the hypothalamic-pituitary adrenal (HPA) axis, glycemia and in vivo intestinal permeability at PND10 and weaning. Methods: At postnatal day (PND) 2, litters were culled to a maximum of 12 pups. From PND2 to 10, Wistar dams and neonatal litters were subjected to either normal housing and nesting conditions (CTL=control litters) with wood shaving bedding and 1 paper towel or LNS with half piece of paper towel placed onto wire bottom flooring. Maternal behavior was assessed in 7 dams/group once daily from PND2 to 9 for 50 min between 9:00-10:00 am. At PND10 and PND21, blood was collected from pups for determination of glucose, plasma corticosterone and in vivo intestinal permeability after oral gavage with fluorescein isothiocyanate-dextran 4kDa (FD4). Results: Dams exposed to LNS showed a significant increase in percentage of time spent in building a nest (118%), self-grooming (69%), and putting the pups back to the nest (167%) compared to CTL. Frequency of pups outside of the nest was 3.7 times higher in LNS than in CTL (2.2 ± 0.6 vs. 0.6 ± 0.2; P < 0.05) while time spent licking and grooming the pups and eating/drinking was not different from that of CTL. At PND10, LNS male and female pups exhibited respectively a reduction of body weight by 4% and 5%, adrenal weights/100g body weight by 17 % and 18%, plasma corticosterone levels by 64% and 62% and blood glucose by 11% and 12% compared to same sex CTL pups. Intestinal permeability was increased only in male LNS pups by 2.7-fold (P <0.001). At PND21, pups from LNS group had similar significant reduction in body weight in both sexes, and hypercorticosteronemia (67 % in males and 147% in females, compared to same sex CTL with females>males). Intestinal permeability to FD4 was increased only in LNS females (1.7-fold compared to CTL, P<0.01). No sex difference was observed in any of the parameters in CTL pups at either PND10 or 21. Conclusions: These data indicate that alterations of maternal behavior induced by LNS delayed the maturation of the HPA axis at PND10 followed by elevated basal corticosteronemia at weaning. The sex-dependent alterations of intestinal epithelial barrier development where male pups are affected earlier than female may have implications in the occurrence of visceral pain later in life.
Figure 1: (A) Histopathology of an EMR tissue, showing regions of dysplasia (yellow), metaplasia (green) and squamous (light blue). Black scale bar 2 mm. (B) Corresponding coregistered ultrasound (greyscale) and photoacoustic images (blue - 750 nm, red - 850 nm), with the corresponding regions of interest (yellow - dysplasia, green - metaplasia). White scale bar 2 mm. (C-F) Frequency histograms of the PAI intensities at 680 (C), 750 (D) and 850 (E) nm, and corresponding calculated relative total hemoglobin (F). In all cases, the mean signal intensity over the regions of interest for dysplasia (yellow in A) is higher than for metaplasia (green in A). Two tailed t-test shows statistically significant differences for all cases.
Su2078 T2 Turbo-Spin-Echo (T2-TSE) MRI for Visceral and Subcutaneous Inflammatory Quantification in Active Celiac and Obese Patients Raffaele Borghini, Francesca Greco, Gabriele Masselli, Carla Lubrano, Marco Di Tola, Giuseppe Donato, Alessandra Piermattei, Antonio Picarelli BACKGROUND: Celiac disease (CD) is a systemic inflammatory disease, involving not only the gastrointestinal tract. It has been recently demonstrated that adipose tissue infiltration by proinflammatory immune cells causes a chronic low-grade inflammation in obese patients. T2-weighted turbo spin-echo (TSE) images at Magnetic Resonance Imaging (MRI) proved to be useful in evaluation of inflammatory states. AIM: Our aim was to evaluate the usefulness of T2-weighted TSE at MRI for differentiating inflammatory alterations in visceral and subcutaneous sites in celiac and obese patients. MATERIALS AND METHODS: 46 total patients have been enrolled and divided in 3 different groups. Group A: 10 celiac patients from our Gastrointestinal Unit, on gluten-containing diet, presenting positive specific antibodies and intestinal villous atrophy. Group B: 16 obese (BMI >30) not-celiac patients from the Endocrine Unit of our Hospital. Group C: 12 blood donors enrolled as healthy nonceliac non-obese controls. MRI was performed by using a 1.5-T MR unit and T2-weighted TSE images of visceral fat were obtained in 4 different regions of interest (ROI) (duodenum, celiac tripod, distal descendending colon and terminal ileum). T2-weighted TSE images of subcutaneous fat were also obtained in two specific ROI (right and left hips). Statistical evaluation of T2 relaxation time was performed by using Kruskal Wallis test after D'AgostinoPearson omnibus test. Afterwards, Dunn's multiple comparison test was performed among the three groups examined for overall p<0.05. Statistical evaluations were performed with
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Clinical Study and Analysis of Ex Vivo Photoacoustic Imaging in Endoscopic Mucosal Resection Tissues in Barrett's Esophagus Liang Lim, Catherine Streutker, Norman E. Marcon, Maria Cirocco, Alexandra Lao, Vladimir Iakovlev, Ralph DaCosta, Brian Wilson Accurate endoscopic detection of dysplasia in patients with Barrett's esophagus (BE) remains a major unmet clinical need. Current diagnosis uses multiple biopsies under endoscopic image guidance, where up to 99% of the tissue remains unsampled, leading to significant risk of missing dysplasia. We have conducted an ex vivo clinical trial using photoacoustic imaging (PAI) in BE patients undergoing endoscopic mucosal resection (EMR) with known high-grade dysplasia, for the purpose of characterizing the esophageal microvascular pattern. The long-term goal is to perform in vivo endoscopic PAI for dysplasia detection and staging by taking advantage of PAI's ability to image through the mucosa into the submucosa. Thirteen (13) tissue samples from 8 patients were analyzed, spanning a range of pathological classifications, including columnar type mucosa (5), dysplasia (5) and intramucosal adenocarcinoma (3). EMR tissues were mounted immediately after resection with the luminal side up on a clear agar slab and covered with ultrasound gel. The PAI transducer (40 MHz center frequency) was placed along the short axis of the tissue. Acoustic image slices (ultrasound and photoacoustic at 680, 750, 824, 850 and 970 nm) were simultaneously acquired covering the full length of the sample, each image slice having a field of view of 14 mm (width) by 15 mm (depth). The tissues were then sliced along the acoustic image slice at approximately 2 mm spacing and fixed in formalin for histopathology with H&E staining. From the acoustic images, we created 3D reconstruction of the full ex vivo tissue volume and generated images of the relative hemoglobin concentrations (oxy-, deoxy- and total) and oxygen saturation. We co-registered the acoustic images and the corresponding histological images, as illustrated in Figures 1A-B. The photoacoustic signal distribution within the tissue appears to coincide with the distribution of blood (dark pink on the H&E images), the main optical absorber in EMR tissue. These initial PAI + ultrasound images have demonstrated the technical feasibility of this approach and point to the potential of PAI to reveal the microvascular pattern within EMR specimens. Analysis of total hemoglobin and the intrinsic acoustic signals (Figure 1C-F) are currently being assessed against histologically diagnosed H&E images for each histopathological classification. These results will be reported and the limitations of this type of photoacoustic mapping of BE will be discussed.
Su2079 Computer Vision Analysis and Content Sampling for Intestinal Organoid Assays Ian A. Williamson, Adam D. Gracz, Scott Magness, Nancy Allbritton 3D intestinal organoid culture allows for real time observation of stem cell driven tissue renewal but impedes repetitive observation of individual cells/organoids in 3D space. Recently, we employed microraft array devices (MRAs) to spatially separating cells in microwells addressed to allow for recall of specific cells/organoids in one of the 15,000 microwells. We optimized CellProfiler computer vision pipelines to reduce user burden by automatically quantifying content and growth data from images of each microwell . First we made pipelines to accurately count the stem and niche cells initially present in each microwell to investigate the effect of cell-signaling interactions on organoid formation. We observed that organoid formation was only significantly increased when stem and niche cells were in physical contact suggesting that secreted factors were not sufficient for niche support. Organoids cultured long-term have widely variable appearances but display two distinct early morphologies discerned by the thickness of their outer monolayer. We developed further pipelines to quantify organoid monolayer width and identify the central pseudolumen compartment. Semi-automated organoid retrieval and downstream gene expression analysis showed significant differences between the two morphologies. Our MRA computer vision technologies will provide a basis for developing fully automated intestinal organoid based assays relating organoid growth/morphology to cell-signaling events.
Su2080 Limited Nesting Stress Alters Maternal Behavior and In Vivo Intestinal Permeability in a Sex-Dependent Manner in Wistar Rat Pups Nabila Moussaoui, Muriel H. Larauche, Mandy Biraud, Jenny Molet, Mulugeta Million, Emeran A. Mayer, Yvette Tache Background: In human, early-life adverse events predispose to psychiatric illness later in life and to functional gastrointestinal disorders such as irritable bowel syndrome. Experimental model of limited nesting stress (LNS) postnatally has been developed to recapitulate elements of human postnatal conditions where the mother is present but chronically provides erratic or unpredictable maternal care in an impoverished environment. Aims: To characterize the alterations of maternal behaviors induced by LNS in Wistar rats and to assess the impact in the offspring on components of the hypothalamic-pituitary adrenal (HPA) axis, glycemia and in vivo intestinal permeability at PND10 and weaning. Methods: At postnatal day (PND) 2, litters were culled to a maximum of 12 pups. From PND2 to 10, Wistar dams and neonatal litters were subjected to either normal housing and nesting conditions (CTL=control litters) with wood shaving bedding and 1 paper towel or LNS with half piece of paper towel placed onto wire bottom flooring. Maternal behavior was assessed in 7 dams/group once daily from PND2 to 9 for 50 min between 9:00-10:00 am. At PND10 and PND21, blood was collected from pups for determination of glucose, plasma corticosterone and in vivo intestinal permeability after oral gavage with fluorescein isothiocyanate-dextran 4kDa (FD4). Results: Dams exposed to LNS showed a significant increase in percentage of time spent in building a nest (118%), self-grooming (69%), and putting the pups back to the nest (167%) compared to CTL. Frequency of pups outside of the nest was 3.7 times higher in LNS than in CTL (2.2 ± 0.6 vs. 0.6 ± 0.2; P < 0.05) while time spent licking and grooming the pups and eating/drinking was not different from that of CTL. At PND10, LNS male and female pups exhibited respectively a reduction of body weight by 4% and 5%, adrenal weights/100g body weight by 17 % and 18%, plasma corticosterone levels by 64% and 62% and blood glucose by 11% and 12% compared to same sex CTL pups. Intestinal permeability was increased only in male LNS pups by 2.7-fold (P <0.001). At PND21, pups from LNS group had similar significant reduction in body weight in both sexes, and hypercorticosteronemia (67 % in males and 147% in females, compared to same sex CTL with females>males). Intestinal permeability to FD4 was increased only in LNS females (1.7-fold compared to CTL, P<0.01). No sex difference was observed in any of the parameters in CTL pups at either PND10 or 21. Conclusions: These data indicate that alterations of maternal behavior induced by LNS delayed the maturation of the HPA axis at PND10 followed by elevated basal corticosteronemia at weaning. The sex-dependent alterations of intestinal epithelial barrier development where male pups are affected earlier than female may have implications in the occurrence of visceral pain later in life.
Figure 1: (A) Histopathology of an EMR tissue, showing regions of dysplasia (yellow), metaplasia (green) and squamous (light blue). Black scale bar 2 mm. (B) Corresponding coregistered ultrasound (greyscale) and photoacoustic images (blue - 750 nm, red - 850 nm), with the corresponding regions of interest (yellow - dysplasia, green - metaplasia). White scale bar 2 mm. (C-F) Frequency histograms of the PAI intensities at 680 (C), 750 (D) and 850 (E) nm, and corresponding calculated relative total hemoglobin (F). In all cases, the mean signal intensity over the regions of interest for dysplasia (yellow in A) is higher than for metaplasia (green in A). Two tailed t-test shows statistically significant differences for all cases.
Su2078 T2 Turbo-Spin-Echo (T2-TSE) MRI for Visceral and Subcutaneous Inflammatory Quantification in Active Celiac and Obese Patients Raffaele Borghini, Francesca Greco, Gabriele Masselli, Carla Lubrano, Marco Di Tola, Giuseppe Donato, Alessandra Piermattei, Antonio Picarelli BACKGROUND: Celiac disease (CD) is a systemic inflammatory disease, involving not only the gastrointestinal tract. It has been recently demonstrated that adipose tissue infiltration by proinflammatory immune cells causes a chronic low-grade inflammation in obese patients. T2-weighted turbo spin-echo (TSE) images at Magnetic Resonance Imaging (MRI) proved to be useful in evaluation of inflammatory states. AIM: Our aim was to evaluate the usefulness of T2-weighted TSE at MRI for differentiating inflammatory alterations in visceral and subcutaneous sites in celiac and obese patients. MATERIALS AND METHODS: 46 total patients have been enrolled and divided in 3 different groups. Group A: 10 celiac patients from our Gastrointestinal Unit, on gluten-containing diet, presenting positive specific antibodies and intestinal villous atrophy. Group B: 16 obese (BMI >30) not-celiac patients from the Endocrine Unit of our Hospital. Group C: 12 blood donors enrolled as healthy nonceliac non-obese controls. MRI was performed by using a 1.5-T MR unit and T2-weighted TSE images of visceral fat were obtained in 4 different regions of interest (ROI) (duodenum, celiac tripod, distal descendending colon and terminal ileum). T2-weighted TSE images of subcutaneous fat were also obtained in two specific ROI (right and left hips). Statistical evaluation of T2 relaxation time was performed by using Kruskal Wallis test after D'AgostinoPearson omnibus test. Afterwards, Dunn's multiple comparison test was performed among the three groups examined for overall p<0.05. Statistical evaluations were performed with
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