Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Deletion of the Cardiomyocyte Glucocorticoid Receptor Leads to Sexually Dimorphic Changes in Cardiac Gene Expression and Progression to Heart Failure

doi: 10.1161/JAHA.118.011012

Figure Lengend Snippet: Ablation of the glucocorticoid receptor in cardiomyocytes leads to an increased mortality rate in male cardio GRKO mice as compared with their female counterparts. Kaplan–Meier analysis of survival after birth was performed for male and female control and cardio GRKO mice. Shown are survival male and female controls and cardio GRKO . Data are from control mice (27 males, 28 females) and cardio GRKO mice (24 males, 23 females). Male controls vs male cardio GRKO . cardioGRKO indicates cardiomyocyte‐specific glucocorticoid receptor knockout. *** P <0.001; female controls vs female cardio GRKO ††† P <0.001; male cardio GRKO vs female cardio GRKO ‡‡‡ P <0.001.

Article Snippet: Adult cardiomyocytes from control and cardioGRKO mice were isolated using a commercially available kit (Perfusion Adumyts Cardiomyocyte Isolation Kit, Cellutron, Baltimore, MD).

Techniques: Control, Knock-Out

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Deletion of the Cardiomyocyte Glucocorticoid Receptor Leads to Sexually Dimorphic Changes in Cardiac Gene Expression and Progression to Heart Failure

doi: 10.1161/JAHA.118.011012

Figure Lengend Snippet: GR mRNA and protein are significantly reduced in the hearts of cardiomyocyte‐specific glucocorticoid receptor knockout (cardio GRKO ) mice and no glucocorticoid receptor ( GR ) is detected in the cardiomyocytes of the knockout mice. A , RT ‐polymerase chain reaction of GR mRNA from hearts of control and cardio GRKO mice. B , Representative immunoblot show GR levels in control ( GR flox) and knockout hearts. Hearts from male cardio GRKO have reduction of ≈70% in GR total protein levels, whereas female hearts present a reduction of ≈80% in GR protein levels. Immunoblots were quantified by densitometry using National Institutes of Health ImageJ analysis software. Data are mean± SEM (n=5 mice per group). * P <0.05; *** P <0.0001. C , Representative immunofluorescence staining of heart sections from control and cardio GRKO mice with anti‐troponin I (green) and anti‐ GR (red) antibodies. DAPI is shown in blue. Data represent n=3 mice per sex and genotype. All pictures were acquired on a Leica TCS SP 5 Spectral Confocal Microscope equipped with a ×40 (oil) objective. CardioGRKO indicates cardiomyocyte‐specific glucocorticoid receptor knockout; CGRKO, cardioGRKO; GR, glucocorticoid receptor.

Article Snippet: Adult cardiomyocytes from control and cardioGRKO mice were isolated using a commercially available kit (Perfusion Adumyts Cardiomyocyte Isolation Kit, Cellutron, Baltimore, MD).

Techniques: Knock-Out, Polymerase Chain Reaction, Control, Western Blot, Software, Immunofluorescence, Staining, Microscopy

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Deletion of the Cardiomyocyte Glucocorticoid Receptor Leads to Sexually Dimorphic Changes in Cardiac Gene Expression and Progression to Heart Failure

doi: 10.1161/JAHA.118.011012

Figure Lengend Snippet: mRNA levels of genes associated with cardiac pathology and the glucocorticoid receptor ( GR ) levels in the heart. Total RNA was isolated from whole hearts of 3‐month‐old male and female control ( GR flox) and cardiomyocyte‐specific glucocorticoid receptor knockout mice. Expression levels of β‐myosin heavy chain ( A ), skeletal muscle α‐actin ( B ), brain natriuretic peptide ( C ), were measured by QRT‐PCR. Data represent n=5 mice per sex and genotype. β‐Mhc indicates β‐myosin heavy chain; cCGRkO, cardiomyocyte‐specific glucocorticoid receptor knockout; GR, glucocorticoid receptor; Nppb , brain natriuretic peptide; Ska , skeletal muscle α‐actin. * P <0.05, and *** P <0.001.

Article Snippet: Adult cardiomyocytes from control and cardioGRKO mice were isolated using a commercially available kit (Perfusion Adumyts Cardiomyocyte Isolation Kit, Cellutron, Baltimore, MD).

Techniques: Isolation, Control, Knock-Out, Expressing, Quantitative RT-PCR

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Deletion of the Cardiomyocyte Glucocorticoid Receptor Leads to Sexually Dimorphic Changes in Cardiac Gene Expression and Progression to Heart Failure

doi: 10.1161/JAHA.118.011012

Figure Lengend Snippet: Pattern of gene expression between male and female hearts from control (glucocorticoid receptor [ GR ] fl/fl) and cardiomyocyte‐specific glucocorticoid receptor knockout mice. A , Heat map of RNA ‐seq data with hierarchical clustering of genes that are differentially expressed between control and cardio GRKO male and female mice. Red indicates high‐expressing genes whereas blue indicates low‐expressing genes. B , Upper panel: Analysis of RNA ‐seq results showing significantly altered genes in each comparison, male GR fl/fl to male cardio GRKO (4262 genes), female GR fl/fl to female cardio GRKO mice (4559 genes), male GR fl/fl to female GR fl/fl (6279 genes), and male cardio GRKO to female cardio GRKO (3879 genes). Lower panel: Venn diagram of the results overlaying male GR fl/fl vs female GR fl/fl (left circle) to male cardio GRKO vs female cardio GRKO mice (right circle). 2299 genes are in common between controls and cardio GRKO hearts (intersection). Cardiomyocyte GR regulates the expression 3980 sexually dimorphic genes in control hearts (64% of a total of 6279 genes are GR dependent). In the absence of GR , 1580 genes are differentially expressed between cardio GRKO male and female hearts. C , Upper right panel: Of the 4262 genes differentially expressed in male cardio GRKO hearts as compared with controls ( GR fl/fl), 306 of these genes are directly or indirectly associated with GR . A similar number of GR associated genes, 392, were found among the 4559 genes with altered expression in the female cardio GRKO hearts. Comparison of these GR ‐associated genes shows that ≈44% of these genes are unique to the cardio GRKO male hearts, while 56% are unique to the female knockout hearts. Upper left panel: Only 60 genes of the 4262 differentially expressed genes in the male cardio GRKO hearts have been related to mineralocorticoid receptor signaling, and only 89 genes of the total of 4559 female knockout dysregulated genes are associated to MR . Venn diagram of these genes overlaying male and female GR fl/fl vs cardio GRKO hearts shows that the majority (100% for males and ≈68% for females) of the reported MR ‐associated genes are common between males and females. Lower panel: About 50% of the MR genes are commonly associated with GR . CardioGRKO indicates cardiomyocyte‐specific glucocorticoid receptor knockout; CGRkO, cardiomyocyte‐specific glucocorticoid receptor knockout; GR, glucocorticoid receptor.

Article Snippet: Adult cardiomyocytes from control and cardioGRKO mice were isolated using a commercially available kit (Perfusion Adumyts Cardiomyocyte Isolation Kit, Cellutron, Baltimore, MD).

Techniques: Gene Expression, Control, Knock-Out, RNA Sequencing, Expressing, Comparison

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Deletion of the Cardiomyocyte Glucocorticoid Receptor Leads to Sexually Dimorphic Changes in Cardiac Gene Expression and Progression to Heart Failure

doi: 10.1161/JAHA.118.011012

Figure Lengend Snippet: Analysis of the glucocorticoid receptor ( GR ) and the mineralocorticoid receptor ( MR ) dependent genes. A , Fifty‐one (male) and 68 (female) of the GR ‐associated genes are related with abnormalities of the heart ventricle. Only 17 and 19 of the MR ‐associated genes dysregulated in cardiomyocyte‐specific glucocorticoid receptor knockout (cardio GRKO hearts were identified within this category (lower panel). Atrial natriuretic peptide (Nppa), brain type natriuretic peptide (Nppb), and cyclooxygenase (Ptgs2) were identified among the common genes. Among the unique MR dependent genes was the vascular endothelial growth factor A (VegfA). Angiotensinogen (Agt) expression seems to be compromised only in male cardio GRKO hearts and its dysregulation is related to alterations in GR signaling. Dysregulation in Agt gene expression is associated with genes involved in cardiac hypertrophy. B , Visualization of the genes connected to Agt in male and female cardio GRKO (upper panel), and network of genes associated with VegfA. Alterations in VegA is associated with genes involved in hypertrophy and angiogenesis. Red depicts induced genes and green represents repressed genes. Orange lines (—) predict activation and blue lines predict inactivation. Yellow lines symbolize gene expression opposite to predicted effect, and gray lines represent that not prediction can be inferred based on the gene expression data. Nppa, Nppb, and Ptgs2 are marked with red asterisk (*). Agt is marked with a purple *, and VegA is marked with an orange *. CardioGRKO indicates cardiomyocyte‐specific glucocorticoid receptor knockout; CGRkO, cardiomyocyte‐specific glucocorticoid receptor knockout; GR, glucocorticoid receptor; MR, mineralocorticoid receptor.

Article Snippet: Adult cardiomyocytes from control and cardioGRKO mice were isolated using a commercially available kit (Perfusion Adumyts Cardiomyocyte Isolation Kit, Cellutron, Baltimore, MD).

Techniques: Knock-Out, Expressing, Gene Expression, Activation Assay

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Deletion of the Cardiomyocyte Glucocorticoid Receptor Leads to Sexually Dimorphic Changes in Cardiac Gene Expression and Progression to Heart Failure

doi: 10.1161/JAHA.118.011012

Figure Lengend Snippet: Effects of deleting cardiomyocyte glucocorticoid receptors on male and female heart gene expression. A , Differentially expressed genes between cardiomyocyte‐specific glucocorticoid receptor knockout (cardio GRKO ) males vs females were analyzed by literature‐based Ingenuity Pathway Analysis ( IPA ) software. The top 13 diseases and biological functions were ranked and assigned an activation z‐score (orange indicates induced; blue indicates repressed). Heart failure was ranked as the most significant activated biological function. One hundred and twenty‐six genes in the heart failure function were significantly altered in male hearts; in contrast, only 77 of these genes were dysregulated in the female hearts. B , Visualization of the genes involved in the heart failure category in male and female cardio GRKO hearts. Genes involved in cardiomyocyte contraction, structure of cardiac sarcomeres, and cardiomyocyte viability are marked with an asterisk (*). ATP ase sarcoplasmic/endoplasmic reticulum Ca 2+ (Atp2a2, Serca), ryanodine receptor 2 (Ryr2), desmin (Des), troponin C1 (Tnnc1), and the solute carrier family 25 member 4 (Slc25a4). Red depicts induced genes and green represents repressed genes. CardioGRKO indicates cardiomyocyte‐specific glucocorticoid receptor knockout.

Article Snippet: Adult cardiomyocytes from control and cardioGRKO mice were isolated using a commercially available kit (Perfusion Adumyts Cardiomyocyte Isolation Kit, Cellutron, Baltimore, MD).

Techniques: Gene Expression, Knock-Out, Software, Activation Assay

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Deletion of the Cardiomyocyte Glucocorticoid Receptor Leads to Sexually Dimorphic Changes in Cardiac Gene Expression and Progression to Heart Failure

doi: 10.1161/JAHA.118.011012

Figure Lengend Snippet: Genes associated with cardiomyocyte contraction, structure of cardiac sarcomeres, and cardiomyocyte viability are dysregulated in male cardiomyocyte‐specific glucocorticoid receptor knockout (cardio GRKO ) hearts but not in their female counterparts. Total RNA was isolated from whole hearts of 3‐month‐old male and female control (glucocorticoid receptor flox/flox) and cardio GRKO mice. A , Expression levels of genes involved in calcium handling ryanodine receptor ( Ryr2 ) and ATP ase sarcoplasmic/endoplasmic reticulum Ca 2+ ( Atp2a2 ). B , Expression levels of genes involved in preserving the structure of cardiac sarcomeres desmin ( Des ) and troponin C1 ( Tnnc1 ). C , Expression level of Slc25a4 (solute carrier family 25 member 4), a mitochondria carrier protein involved in cardiomyocyte viability. Data are mean± SEM (n=4–5 mice per group). * P <0.05, and ** P <0.01. CardioGRKO indicates cardiomyocyte‐specific glucocorticoid receptor knockout.

Article Snippet: Adult cardiomyocytes from control and cardioGRKO mice were isolated using a commercially available kit (Perfusion Adumyts Cardiomyocyte Isolation Kit, Cellutron, Baltimore, MD).

Techniques: Knock-Out, Isolation, Control, Expressing, Preserving

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Deletion of the Cardiomyocyte Glucocorticoid Receptor Leads to Sexually Dimorphic Changes in Cardiac Gene Expression and Progression to Heart Failure

doi: 10.1161/JAHA.118.011012

Figure Lengend Snippet: Expression and quantification of RyR2 (ryanodine receptor 2) protein with altered calcium responses in male cardiomyocyte‐specific glucocorticoid receptor knockout (cardio GRKO ) mice. A , Representative immunoblot show Ryr2 protein levels in control (glucocorticoid receptor flox) and knockout hearts. Hearts from male cardio GRKO have reduction of ≈50% in Ryr2 total protein levels, whereas female hearts do not present a significant reduction in Ryr2 protein levels. Immunoblots were quantified by densitometry using National Institutes of Health ImageJ analysis software. Data are mean± SEM (n=3 mice per group). * P <0.05. B , Intracellular calcium changes were monitored using a calcium sensitive indicator, fluo‐4 in adult cardiomyocytes isolated from control and cardio GRKO male and female mice. Changes in intracellular calcium were monitored before and after application of a low concentration of caffeine (0.2 mmol/L, red arrow) and high concentration of caffeine (10 mmol/L, blue arrow) and are represented as the change in fluo‐4 fluorescence emission intensity. Upper panel graphs are representative data from different cardiomyocytes isolated from control mice. Lower panel graphs are representative data from different cardiomyocytes isolated from cardio GRKO mice. Cardiomyocytes displaying spontaneous intracellular calcium changes in the form of local calcium transients were selected for study. Intracellular calcium changes were monitored at the single cell level with data collected from a region of interest representing an individual cell. Typically, 4 regions of interest were monitored per experiment. CGRKO indicates cardiomyocyte‐specific glucocorticoid receptor knockout; GR, glucocorticoid receptor; Ryr2, ryanodine receptor 2.

Article Snippet: Adult cardiomyocytes from control and cardioGRKO mice were isolated using a commercially available kit (Perfusion Adumyts Cardiomyocyte Isolation Kit, Cellutron, Baltimore, MD).

Techniques: Expressing, Knock-Out, Western Blot, Control, Software, Isolation, Concentration Assay, Fluorescence

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Deletion of the Cardiomyocyte Glucocorticoid Receptor Leads to Sexually Dimorphic Changes in Cardiac Gene Expression and Progression to Heart Failure

doi: 10.1161/JAHA.118.011012

Figure Lengend Snippet: Cardio GRKO male and female mice develop spontaneous left ventricular remodeling. A , Representative images of intact hearts from 6‐month‐old male and female control and cardio GRKO mice. B , Longitudinal hematoxylin & eosin‐stained heart sections from 6‐month‐old male and female control and cardio GRKO mice. Arrows (→) point out to increased left ventricle size and the presence of a thrombus in the left atria of the female cardio GRKO heart. The images are representative of 5 mice per group. CardioGRKO indicates cardiomyocyte‐specific glucocorticoid receptor knockout.

Article Snippet: Adult cardiomyocytes from control and cardioGRKO mice were isolated using a commercially available kit (Perfusion Adumyts Cardiomyocyte Isolation Kit, Cellutron, Baltimore, MD).

Techniques: Control, Staining, Knock-Out

Journal:

Article Title: Evidence that the acute phase of ischemic preconditioning does not require signaling by the A 2B adenosine receptor

doi: 10.1016/j.yjmcc.2010.08.015

Figure Lengend Snippet: LV functional recovery data at 45 min of reperfusion (A–D) and cTnI release data (E) from the isolated mouse hearts studies of IPC. Functional data are expressed as a percentage of baseline values. Refer to Fig. 1 for a detailed description of the experimental protocol. IR, non-preconditioned control group; IPC, preconditioned group. *P<0.05 versus the respective non-preconditioned control group (IR). n = 8–10 hearts/group.

Article Snippet: During the reperfusion period, the coronary perfusate was collected and stored at −80° C until assayed for cardiac troponin I (cTnI) content by radioimmunoassay (Life Diagnostics, Inc., West Chester, PA).

Techniques: Functional Assay, Isolation, Control

Journal: Ontario Health Technology Assessment Series

Article Title: Portable Normothermic Cardiac Perfusion System in Donation After Cardiocirculatory Death: A Health Technology Assessment

doi:

Figure Lengend Snippet: Survival and Deaths Following Cardiac Transplants With Donation After Cardiocirculatory Death Hearts Versus Donation After Neurological Determination of Death Hearts

Article Snippet: Inclusion Criteria English-language full-text publications Studies published since January 1, 1998 Comparative randomized and non-randomized studies or non-comparative studies for effectiveness outcomes; case reports for adverse events/effects outcomes De novo adult heart transplant recipients of donation after cardiocirculatory death (DCD) Portable normothermic cardiac perfusion system (e.g., TransMedics, Inc.

Techniques: Infection

Journal: Ontario Health Technology Assessment Series

Article Title: Portable Normothermic Cardiac Perfusion System in Donation After Cardiocirculatory Death: A Health Technology Assessment

doi:

Figure Lengend Snippet: Survival of Donation After Cardiocirculatory Death Heart Transplant Recipients by Procurement Technique Subgroup

Article Snippet: Inclusion Criteria English-language full-text publications Studies published since January 1, 1998 Comparative randomized and non-randomized studies or non-comparative studies for effectiveness outcomes; case reports for adverse events/effects outcomes De novo adult heart transplant recipients of donation after cardiocirculatory death (DCD) Portable normothermic cardiac perfusion system (e.g., TransMedics, Inc.

Techniques:

Journal: Ontario Health Technology Assessment Series

Article Title: Portable Normothermic Cardiac Perfusion System in Donation After Cardiocirculatory Death: A Health Technology Assessment

doi:

Figure Lengend Snippet: Post-operative Outcomes and Graft Function After Heart Transplant With Either Donation After Cardiocirculatory Death Hearts Using a Portable Normothermic Cardiac Perfusion System or Donation After Neurological Determination of Death Hearts

Article Snippet: Inclusion Criteria English-language full-text publications Studies published since January 1, 1998 Comparative randomized and non-randomized studies or non-comparative studies for effectiveness outcomes; case reports for adverse events/effects outcomes De novo adult heart transplant recipients of donation after cardiocirculatory death (DCD) Portable normothermic cardiac perfusion system (e.g., TransMedics, Inc.

Techniques:

Journal: Ontario Health Technology Assessment Series

Article Title: Portable Normothermic Cardiac Perfusion System in Donation After Cardiocirculatory Death: A Health Technology Assessment

doi:

Figure Lengend Snippet: Ischemic Time for Donation After Cardiocirculatory Death Hearts, by Procurement Technique

Article Snippet: Inclusion Criteria English-language full-text publications Studies published since January 1, 1998 Comparative randomized and non-randomized studies or non-comparative studies for effectiveness outcomes; case reports for adverse events/effects outcomes De novo adult heart transplant recipients of donation after cardiocirculatory death (DCD) Portable normothermic cardiac perfusion system (e.g., TransMedics, Inc.

Techniques: Functional Assay

Journal: Ontario Health Technology Assessment Series

Article Title: Portable Normothermic Cardiac Perfusion System in Donation After Cardiocirculatory Death: A Health Technology Assessment

doi:

Figure Lengend Snippet: Results of Economic Literature Review—Summary

Article Snippet: Inclusion Criteria English-language full-text publications Studies published since January 1, 1998 Comparative randomized and non-randomized studies or non-comparative studies for effectiveness outcomes; case reports for adverse events/effects outcomes De novo adult heart transplant recipients of donation after cardiocirculatory death (DCD) Portable normothermic cardiac perfusion system (e.g., TransMedics, Inc.

Techniques: Control

Journal: Ontario Health Technology Assessment Series

Article Title: Portable Normothermic Cardiac Perfusion System in Donation After Cardiocirculatory Death: A Health Technology Assessment

doi:

Figure Lengend Snippet: Volume of Intervention

Article Snippet: Inclusion Criteria English-language full-text publications Studies published since January 1, 1998 Comparative randomized and non-randomized studies or non-comparative studies for effectiveness outcomes; case reports for adverse events/effects outcomes De novo adult heart transplant recipients of donation after cardiocirculatory death (DCD) Portable normothermic cardiac perfusion system (e.g., TransMedics, Inc.

Techniques: Preserving

Journal: Ontario Health Technology Assessment Series

Article Title: Portable Normothermic Cardiac Perfusion System in Donation After Cardiocirculatory Death: A Health Technology Assessment

doi:

Figure Lengend Snippet: Expected Number of Heart Transplants by Donor Type

Article Snippet: Inclusion Criteria English-language full-text publications Studies published since January 1, 1998 Comparative randomized and non-randomized studies or non-comparative studies for effectiveness outcomes; case reports for adverse events/effects outcomes De novo adult heart transplant recipients of donation after cardiocirculatory death (DCD) Portable normothermic cardiac perfusion system (e.g., TransMedics, Inc.

Techniques: Preserving