The Experts below are selected from a list of 1056 Experts worldwide ranked by ideXlab platform
Robert D Bennett - One of the best experts on this subject based on the ideXlab platform.
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single cell transcriptional profiling of cells derived from regenerating Alveolar Ducts
Frontiers in Medicine, 2020Co-Authors: Alexandra B Ysasi, Akira Tsuda, Robert D Bennett, Willi L Wagner, Cristian D Valenzuela, Andrew B Servais, Saumyadipta Pyne, Jonna Grimsby, Prapti Pokharel, Kenneth J LivakAbstract:Lung regeneration occurs in a variety of adult mammals after surgical removal of one lung (pneumonectomy). Previous studies of murine post-pneumonectomy lung growth have identified regenerative "hotspots" in subpleural Alveolar Ducts; however, the cell-types participating in this process remain unclear. To identify the single cells participating in post-pneumonectomy lung growth, we used laser microdissection, enzymatic digestion and microfluidic isolation. Single-cell transcriptional analysis of the murine Alveolar Duct cells was performed using the C1 integrated fluidic circuit (Fluidigm) and a custom PCR panel designed for lung growth and repair genes. The multi-dimensional data set was analyzed using visualization software based on the tSNE algorithm. The analysis identified 6 cell clusters; 1 cell cluster was present only after pneumonectomy. This post-pneumonectomy cluster was significantly less transcriptionally active than 3 other clusters and may represent a transitional cell population. A provisional cluster identity for 4 of the 6 cell clusters was obtained by embedding bulk transcriptional data into the tSNE analysis. The transcriptional pattern of the 6 clusters was further analyzed for genes associated with lung repair, matrix proDuction, and angiogenesis. The data demonstrated that multiple cell-types (clusters) transcribed genes linked to these basic functions. We conclude that the coordinated gene expression across multiple cell clusters is likely a response to a shared regenerative microenvironment within the subpleural Alveolar Ducts.
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structural remodeling of the post pneumonectomy lung is characterized by septal retraction and Alveolar Duct dilation
The FASEB Journal, 2015Co-Authors: Willi L Wagner, Akira Tsuda, Alexandra B Ysasi, Robert D Bennett, Cristian D Valenzuela, Moritz A Konerding, Maximilian Ackermann, Steven J MentzerAbstract:Murine pneumonectomy (PNX) is an established model of adult compensatory lung growth, involving structural remodeling of the peripheral parenchyma. Following left PNX, positron emission tomography and computerized tomography have demonstrated heterogeneous growth patterns within the remaining right lung. To characterize the structural changes associated with these observations, we analyzed histological sections of the right lung between 3 and 22 days after PNX. Within 3 days of PNX, Alveolar septae were significantly shorter (p<0.05) in a majority of Alveolar Ducts, while septal angle remained unchanged. Septal retraction resulted in an apparent dilation of the Alveolar Duct with a significantly decreased Alveolar Duct diameter ratio (Dout:Din; p<0.01). The greatest percentage of dilated Alveolar Ducts was seen at day 3, with 46% of Alveolar Ducts in the most dilated Alveolar Duct ratio quintile. Heterogeneous septal lengthening was observed between 4 and 22 days after PNX, with average septal length retu...
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laser microdissection of the Alveolar Duct enables single cell genomic analysis
Frontiers in Oncology, 2014Co-Authors: Robert D Bennett, Alexandra B Ysasi, Willi L Wagner, Saumyadipta Pyne, Janeil Belle, Moritz A Konerding, Paul C Blainey, Steven J MentzerAbstract:Complex tissues such as the lung are composed of structural hierarchies such as alveoli, Alveolar Ducts and lobules. Some structural units, such as the Alveolar Duct, appear to selectively participate in tissue regeneration. Here, we demonstrate an approach to conDuct laser microdissection of the lung Alveolar Duct for single-cell PCR analysis. Our approach involved three steps. 1) The initial preparation used mechanical sectioning of the lung tissue with sufficient thickness to encompass the structure of interest. In the case of the Alveolar Duct, the precision-cut lung slices were 200um thick; the slices were processed using near-physiologic conditions to preserve the state of viable cells. 2) The lung slices were examined by transmission light microscopy to target the Alveolar Duct. The air-filled lung was sufficiently accessible by light microscopy that counterstains or fluorescent labels were unnecessary to identify the Alveolar Duct. 3) The enzymatic and microfluidic isolation of single cells allowed for the harvest of as few as several thousand cells for PCR analysis. Microfluidics based arrays were used to measure the expression of selected marker genes in individual cells to characterize different cell populations. Preliminary work suggests the unique value of this approach to understanding the intra- and intercellular interactions within the regenerating Alveolar Duct.
Alexandra B Ysasi - One of the best experts on this subject based on the ideXlab platform.
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single cell transcriptional profiling of cells derived from regenerating Alveolar Ducts
Frontiers in Medicine, 2020Co-Authors: Alexandra B Ysasi, Akira Tsuda, Robert D Bennett, Willi L Wagner, Cristian D Valenzuela, Andrew B Servais, Saumyadipta Pyne, Jonna Grimsby, Prapti Pokharel, Kenneth J LivakAbstract:Lung regeneration occurs in a variety of adult mammals after surgical removal of one lung (pneumonectomy). Previous studies of murine post-pneumonectomy lung growth have identified regenerative "hotspots" in subpleural Alveolar Ducts; however, the cell-types participating in this process remain unclear. To identify the single cells participating in post-pneumonectomy lung growth, we used laser microdissection, enzymatic digestion and microfluidic isolation. Single-cell transcriptional analysis of the murine Alveolar Duct cells was performed using the C1 integrated fluidic circuit (Fluidigm) and a custom PCR panel designed for lung growth and repair genes. The multi-dimensional data set was analyzed using visualization software based on the tSNE algorithm. The analysis identified 6 cell clusters; 1 cell cluster was present only after pneumonectomy. This post-pneumonectomy cluster was significantly less transcriptionally active than 3 other clusters and may represent a transitional cell population. A provisional cluster identity for 4 of the 6 cell clusters was obtained by embedding bulk transcriptional data into the tSNE analysis. The transcriptional pattern of the 6 clusters was further analyzed for genes associated with lung repair, matrix proDuction, and angiogenesis. The data demonstrated that multiple cell-types (clusters) transcribed genes linked to these basic functions. We conclude that the coordinated gene expression across multiple cell clusters is likely a response to a shared regenerative microenvironment within the subpleural Alveolar Ducts.
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structural remodeling of the post pneumonectomy lung is characterized by septal retraction and Alveolar Duct dilation
The FASEB Journal, 2015Co-Authors: Willi L Wagner, Akira Tsuda, Alexandra B Ysasi, Robert D Bennett, Cristian D Valenzuela, Moritz A Konerding, Maximilian Ackermann, Steven J MentzerAbstract:Murine pneumonectomy (PNX) is an established model of adult compensatory lung growth, involving structural remodeling of the peripheral parenchyma. Following left PNX, positron emission tomography and computerized tomography have demonstrated heterogeneous growth patterns within the remaining right lung. To characterize the structural changes associated with these observations, we analyzed histological sections of the right lung between 3 and 22 days after PNX. Within 3 days of PNX, Alveolar septae were significantly shorter (p<0.05) in a majority of Alveolar Ducts, while septal angle remained unchanged. Septal retraction resulted in an apparent dilation of the Alveolar Duct with a significantly decreased Alveolar Duct diameter ratio (Dout:Din; p<0.01). The greatest percentage of dilated Alveolar Ducts was seen at day 3, with 46% of Alveolar Ducts in the most dilated Alveolar Duct ratio quintile. Heterogeneous septal lengthening was observed between 4 and 22 days after PNX, with average septal length retu...
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laser microdissection of the Alveolar Duct enables single cell genomic analysis
Frontiers in Oncology, 2014Co-Authors: Robert D Bennett, Alexandra B Ysasi, Willi L Wagner, Saumyadipta Pyne, Janeil Belle, Moritz A Konerding, Paul C Blainey, Steven J MentzerAbstract:Complex tissues such as the lung are composed of structural hierarchies such as alveoli, Alveolar Ducts and lobules. Some structural units, such as the Alveolar Duct, appear to selectively participate in tissue regeneration. Here, we demonstrate an approach to conDuct laser microdissection of the lung Alveolar Duct for single-cell PCR analysis. Our approach involved three steps. 1) The initial preparation used mechanical sectioning of the lung tissue with sufficient thickness to encompass the structure of interest. In the case of the Alveolar Duct, the precision-cut lung slices were 200um thick; the slices were processed using near-physiologic conditions to preserve the state of viable cells. 2) The lung slices were examined by transmission light microscopy to target the Alveolar Duct. The air-filled lung was sufficiently accessible by light microscopy that counterstains or fluorescent labels were unnecessary to identify the Alveolar Duct. 3) The enzymatic and microfluidic isolation of single cells allowed for the harvest of as few as several thousand cells for PCR analysis. Microfluidics based arrays were used to measure the expression of selected marker genes in individual cells to characterize different cell populations. Preliminary work suggests the unique value of this approach to understanding the intra- and intercellular interactions within the regenerating Alveolar Duct.
Steven J Mentzer - One of the best experts on this subject based on the ideXlab platform.
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structural remodeling of the post pneumonectomy lung is characterized by septal retraction and Alveolar Duct dilation
The FASEB Journal, 2015Co-Authors: Willi L Wagner, Akira Tsuda, Alexandra B Ysasi, Robert D Bennett, Cristian D Valenzuela, Moritz A Konerding, Maximilian Ackermann, Steven J MentzerAbstract:Murine pneumonectomy (PNX) is an established model of adult compensatory lung growth, involving structural remodeling of the peripheral parenchyma. Following left PNX, positron emission tomography and computerized tomography have demonstrated heterogeneous growth patterns within the remaining right lung. To characterize the structural changes associated with these observations, we analyzed histological sections of the right lung between 3 and 22 days after PNX. Within 3 days of PNX, Alveolar septae were significantly shorter (p<0.05) in a majority of Alveolar Ducts, while septal angle remained unchanged. Septal retraction resulted in an apparent dilation of the Alveolar Duct with a significantly decreased Alveolar Duct diameter ratio (Dout:Din; p<0.01). The greatest percentage of dilated Alveolar Ducts was seen at day 3, with 46% of Alveolar Ducts in the most dilated Alveolar Duct ratio quintile. Heterogeneous septal lengthening was observed between 4 and 22 days after PNX, with average septal length retu...
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laser microdissection of the Alveolar Duct enables single cell genomic analysis
Frontiers in Oncology, 2014Co-Authors: Robert D Bennett, Alexandra B Ysasi, Willi L Wagner, Saumyadipta Pyne, Janeil Belle, Moritz A Konerding, Paul C Blainey, Steven J MentzerAbstract:Complex tissues such as the lung are composed of structural hierarchies such as alveoli, Alveolar Ducts and lobules. Some structural units, such as the Alveolar Duct, appear to selectively participate in tissue regeneration. Here, we demonstrate an approach to conDuct laser microdissection of the lung Alveolar Duct for single-cell PCR analysis. Our approach involved three steps. 1) The initial preparation used mechanical sectioning of the lung tissue with sufficient thickness to encompass the structure of interest. In the case of the Alveolar Duct, the precision-cut lung slices were 200um thick; the slices were processed using near-physiologic conditions to preserve the state of viable cells. 2) The lung slices were examined by transmission light microscopy to target the Alveolar Duct. The air-filled lung was sufficiently accessible by light microscopy that counterstains or fluorescent labels were unnecessary to identify the Alveolar Duct. 3) The enzymatic and microfluidic isolation of single cells allowed for the harvest of as few as several thousand cells for PCR analysis. Microfluidics based arrays were used to measure the expression of selected marker genes in individual cells to characterize different cell populations. Preliminary work suggests the unique value of this approach to understanding the intra- and intercellular interactions within the regenerating Alveolar Duct.
Willi L Wagner - One of the best experts on this subject based on the ideXlab platform.
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single cell transcriptional profiling of cells derived from regenerating Alveolar Ducts
Frontiers in Medicine, 2020Co-Authors: Alexandra B Ysasi, Akira Tsuda, Robert D Bennett, Willi L Wagner, Cristian D Valenzuela, Andrew B Servais, Saumyadipta Pyne, Jonna Grimsby, Prapti Pokharel, Kenneth J LivakAbstract:Lung regeneration occurs in a variety of adult mammals after surgical removal of one lung (pneumonectomy). Previous studies of murine post-pneumonectomy lung growth have identified regenerative "hotspots" in subpleural Alveolar Ducts; however, the cell-types participating in this process remain unclear. To identify the single cells participating in post-pneumonectomy lung growth, we used laser microdissection, enzymatic digestion and microfluidic isolation. Single-cell transcriptional analysis of the murine Alveolar Duct cells was performed using the C1 integrated fluidic circuit (Fluidigm) and a custom PCR panel designed for lung growth and repair genes. The multi-dimensional data set was analyzed using visualization software based on the tSNE algorithm. The analysis identified 6 cell clusters; 1 cell cluster was present only after pneumonectomy. This post-pneumonectomy cluster was significantly less transcriptionally active than 3 other clusters and may represent a transitional cell population. A provisional cluster identity for 4 of the 6 cell clusters was obtained by embedding bulk transcriptional data into the tSNE analysis. The transcriptional pattern of the 6 clusters was further analyzed for genes associated with lung repair, matrix proDuction, and angiogenesis. The data demonstrated that multiple cell-types (clusters) transcribed genes linked to these basic functions. We conclude that the coordinated gene expression across multiple cell clusters is likely a response to a shared regenerative microenvironment within the subpleural Alveolar Ducts.
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structural remodeling of the post pneumonectomy lung is characterized by septal retraction and Alveolar Duct dilation
The FASEB Journal, 2015Co-Authors: Willi L Wagner, Akira Tsuda, Alexandra B Ysasi, Robert D Bennett, Cristian D Valenzuela, Moritz A Konerding, Maximilian Ackermann, Steven J MentzerAbstract:Murine pneumonectomy (PNX) is an established model of adult compensatory lung growth, involving structural remodeling of the peripheral parenchyma. Following left PNX, positron emission tomography and computerized tomography have demonstrated heterogeneous growth patterns within the remaining right lung. To characterize the structural changes associated with these observations, we analyzed histological sections of the right lung between 3 and 22 days after PNX. Within 3 days of PNX, Alveolar septae were significantly shorter (p<0.05) in a majority of Alveolar Ducts, while septal angle remained unchanged. Septal retraction resulted in an apparent dilation of the Alveolar Duct with a significantly decreased Alveolar Duct diameter ratio (Dout:Din; p<0.01). The greatest percentage of dilated Alveolar Ducts was seen at day 3, with 46% of Alveolar Ducts in the most dilated Alveolar Duct ratio quintile. Heterogeneous septal lengthening was observed between 4 and 22 days after PNX, with average septal length retu...
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laser microdissection of the Alveolar Duct enables single cell genomic analysis
Frontiers in Oncology, 2014Co-Authors: Robert D Bennett, Alexandra B Ysasi, Willi L Wagner, Saumyadipta Pyne, Janeil Belle, Moritz A Konerding, Paul C Blainey, Steven J MentzerAbstract:Complex tissues such as the lung are composed of structural hierarchies such as alveoli, Alveolar Ducts and lobules. Some structural units, such as the Alveolar Duct, appear to selectively participate in tissue regeneration. Here, we demonstrate an approach to conDuct laser microdissection of the lung Alveolar Duct for single-cell PCR analysis. Our approach involved three steps. 1) The initial preparation used mechanical sectioning of the lung tissue with sufficient thickness to encompass the structure of interest. In the case of the Alveolar Duct, the precision-cut lung slices were 200um thick; the slices were processed using near-physiologic conditions to preserve the state of viable cells. 2) The lung slices were examined by transmission light microscopy to target the Alveolar Duct. The air-filled lung was sufficiently accessible by light microscopy that counterstains or fluorescent labels were unnecessary to identify the Alveolar Duct. 3) The enzymatic and microfluidic isolation of single cells allowed for the harvest of as few as several thousand cells for PCR analysis. Microfluidics based arrays were used to measure the expression of selected marker genes in individual cells to characterize different cell populations. Preliminary work suggests the unique value of this approach to understanding the intra- and intercellular interactions within the regenerating Alveolar Duct.
Annick Clement - One of the best experts on this subject based on the ideXlab platform.
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surfactant protein c gene sftpc mutation associated lung disease high resolution computed tomography hrct findings and its relation to histological analysis
Pediatric Pulmonology, 2010Co-Authors: M Mechri, Ralph Epaud, Sophie Emond, Aurore Coulomb, Francis Jaubert, A Tarrant, Delphine Feldmann, Florence Flamein, Annick ClementAbstract:Aim of the Study Determine high-resolution tomography (HRCT) scan characteristics in children with SFTPC mutation and correlate them to histological findings. Patients and Methods This retrospective multicenter study included 15 children (7 females and 8 males) with SFTPC mutations. HRCT scans have been performed in all the children and lung biopsies in 8 children. Results From all signs assessed on initial HRCT scans, ground-glass opacities (n = 14, 93%) and lung cysts (n = 6, 40%) were predominant. Interlobular septal thickening (n = 1, 7%), air space consolidation (n = 1, 7%), paraseptal emphysema (n = 2, 13%), and pulmonary nodules (n = 1, 7%) were also found. Histological analysis revealed accumulation of macrophages in the Alveolar lumen, type II pneumocyte hyperplasia, and Alveolar septal thickening. Dilatation of the respiratory bronchiole and Alveolar Duct associated with muscular hyperplasia were also described. Interestingly, lung cysts on HRCT scans were associated with dilatation of terminal bronchioli and Alveolar Duct in lung biopsies. Conclusion In children with SFTPC mutations, HRCT scan finding was highly correlated to the histological findings and, as such, represent a useful tool to identify patients that may require SFTPC gene sequencing. Pediatr Pulmonol. 2010; 45:1021–1029. © 2010 Wiley-Liss, Inc.