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1.
Sci Rep ; 14(1): 9991, 2024 05 01.
Article in English | MEDLINE | ID: mdl-38693202

ABSTRACT

Endothelial cells (ECs) have essential roles in cardiac tissue repair after myocardial infarction (MI). To establish stage-specific and long-term effects of the ischemic injury on cardiac ECs, we analyzed their transcriptome at landmark time points after MI in mice. We found that early EC response at Day 2 post-MI centered on metabolic changes, acquisition of proinflammatory phenotypes, initiation of the S phase of cell cycle, and activation of stress-response pathways, followed by progression to mitosis (M/G2 phase) and acquisition of proangiogenic and mesenchymal properties during scar formation at Day 7. In contrast, genes involved in vascular physiology and maintenance of vascular tone were suppressed. Importantly, ECs did not return to pre-injury phenotypes after repair has been completed but maintained inflammatory, fibrotic and thrombotic characteristics and lost circadian rhythmicity. We discovered that the highest induced transcript is the mammalian-specific Sh2d5 gene that promoted migration and invasion of ECs through Rac1 GTPase. Our results revealed a synchronized, temporal activation of disease phenotypes, metabolic pathways, and proliferation in quiescent ECs after MI, indicating that precisely-timed interventions are necessary to optimize cardiac tissue repair and improve outcomes. Furthermore, long-term effects of acute ischemic injury on ECs may contribute to vascular dysfunction and development of heart failure.


Subject(s)
Endothelial Cells , Gene Expression Profiling , Myocardial Infarction , Animals , Myocardial Infarction/genetics , Myocardial Infarction/metabolism , Myocardial Infarction/pathology , Mice , Endothelial Cells/metabolism , Endothelial Cells/pathology , Transcriptome , Male , Mice, Inbred C57BL , Myocardium/metabolism , Myocardium/pathology , rac1 GTP-Binding Protein/metabolism , rac1 GTP-Binding Protein/genetics , Disease Models, Animal , Cell Proliferation , Cell Movement/genetics
2.
Org Biomol Chem ; 19(5): 1133-1144, 2021 02 11.
Article in English | MEDLINE | ID: mdl-33443507

ABSTRACT

The development of new approaches toward chemo- and regioselective functionalization of polycyclic aromatic hydrocarbon (PAH) scaffolds will provide opportunities for the synthesis of novel biologically active small molecules that exploit the high degree of lipophilicity imparted by the PAH unit. Herein, we report a new synthetic method for C-X bond substitution that is speculated to operate via a N-centered radical (NCR) mechanism according to experimental observations. A series of PAH sulfonamides have been synthesized and their biological activity has been evaluated against Gram-negative and Gram-positive bacterial strains (using a BacTiter-Glo assay) along with a series of mammalian cell lines (using CellTiter-Blue and CellTiter-Glo assays). The viability assays have resulted in the discovery of a number of bactericidal compounds that exhibit potency similar to other well-known antibacterials such as kanamycin and tetracycline, along with the discovery of a luciferase inhibitor. Additionally, the physicochemical and drug-likeness properties of the compounds were determined experimentally and using in silico approaches and the results are presented and discussed within.


Subject(s)
Acetates/chemistry , Iodine/chemistry , Iodobenzenes/chemistry , Polycyclic Aromatic Hydrocarbons/chemistry , Sulfonamides/chemical synthesis , Sulfonamides/pharmacology , Anti-Bacterial Agents/chemical synthesis , Anti-Bacterial Agents/chemistry , Anti-Bacterial Agents/pharmacology , Antineoplastic Agents/chemical synthesis , Antineoplastic Agents/chemistry , Antineoplastic Agents/pharmacology , Catalysis , Cell Line, Tumor , Chemistry Techniques, Synthetic , Humans , Microbial Sensitivity Tests , Structure-Activity Relationship , Sulfonamides/chemistry
3.
Int J Mol Sci ; 21(20)2020 Oct 10.
Article in English | MEDLINE | ID: mdl-33050457

ABSTRACT

Brain-derived neurotrophic factor (BDNF) is a neuronal growth and survival factor that harbors cardioprotective qualities that may attenuate dilated cardiomyopathy. In ~30% of the population, BDNF has a common, nonsynonymous single nucleotide polymorphism rs6265 (Val66Met), which might be correlated with increased risk of cardiovascular events. We previously showed that BDNF correlates with better cardiac function in Duchenne muscular dystrophy (DMD) patients. However, the effect of the Val66Met polymorphism on cardiac function has not been determined. The goal of the current study was to determine the effects of rs6265 on BDNF biomarker suitability and DMD cardiac functions more generally. We assessed cardiovascular and skeletal muscle function in human DMD patients segregated by polymorphic allele. We also compared echocardiographic, electrophysiologic, and cardiomyocyte contractility in C57/BL-6 wild-type mice with rs6265 polymorphism and in mdx/mTR (mDMD) mouse model of DMD. In human DMD patients, plasma BDNF levels had a positive correlation with left ventricular function, opposite to that seen in rs6265 carriers. There was also a substantial decrease in skeletal muscle function in carriers compared to the Val homozygotes. Surprisingly, the opposite was true when cardiac function of DMD carriers and non-carriers were compared. On the other hand, Val66Met wild-type mice had only subtle functional differences at baseline but significantly decreased cardiomyocyte contractility. Our results indicate that the Val66Met polymorphism alters myocyte contractility, conferring worse skeletal muscle function but better cardiac function in DMD patients. Moreover, these results suggest a mechanism for the relative preservation of cardiac tissues compared to skeletal muscle in DMD patients and underscores the complexity of BDNF signaling in response to mechanical workload.


Subject(s)
Brain-Derived Neurotrophic Factor/genetics , Cardiomyopathy, Dilated/etiology , Cardiomyopathy, Dilated/metabolism , Genetic Predisposition to Disease , Myocytes, Cardiac/metabolism , Polymorphism, Single Nucleotide , Animals , Cardiomyopathy, Dilated/diagnosis , Cardiomyopathy, Dilated/physiopathology , Disease Models, Animal , Echocardiography , Electrocardiography , Gene Expression Regulation , Genetic Association Studies , Humans , Mice , Mice, Transgenic , Myocardial Contraction
4.
Cardiovasc Res ; 115(2): 395-408, 2019 02 01.
Article in English | MEDLINE | ID: mdl-30169632

ABSTRACT

Aims: Macrophage phagocytosis of dead cells is a prerequisite for inflammation resolution. Because CXCL4 induces macrophage phagocytosis in vitro, we examined the impact of exogenous CXCL4 infusion on cardiac wound healing and macrophage phagocytosis following myocardial infarction (MI). Methods and results: CXCL4 expression significantly increased in the infarct region beginning at Day 3 post-MI, and macrophages were the predominant source. Adult male C57BL/6J mice were subjected to coronary artery occlusion, and MI mice were randomly infused with recombinant mouse CXCL4 or saline beginning at 24 h post-MI by mini-pump infusion. Compared with saline controls, CXCL4 infusion dramatically reduced 7 day post-MI survival [10% (3/30) for CXCL4 vs. 47% (7/15) for saline, P < 0.05] as a result of acute congestive heart failure. By echocardiography, CXCL4 significantly increased left ventricular (LV) volumes and dimensions at Day 5 post-MI (all P < 0.05), despite similar infarct areas compared with saline controls. While macrophage numbers were similar at Day 5 post-MI, CXCL4 infusion increased Ccr4 and Itgb4 and decreased Adamts8 gene levels in the infarct region, all of which linked to CXCL4-mediated cardiac dilation. Isolated Day 5 post-MI macrophages exhibited comparable levels of M1 and M4 markers between saline and CXCL4 groups. Interestingly, by both ex vivo and in vitro phagocytosis assays, CXCL4 reduced macrophage phagocytic capacity, which was connected to decreased levels of the phagocytosis receptor CD36. In vitro, a CD36 neutralizing antibody (CD36Ab) significantly inhibited macrophage phagocytic capacity. The combination of CXCL4 and CD36Ab did not have an additive effect, indicating that CXCL4 regulated phagocytosis through CD36 signalling. CXCL4 infusion significantly elevated infarct matrix metalloproteinase (MMP)-9 levels at Day 5 post-MI, and MMP-9 can cleave CD36 as a down-regulation mechanism. Conclusion: CXCL4 infusion impaired macrophage phagocytic capacity by reducing CD36 levels through MMP-9 dependent and independent signalling, leading to higher mortality and LV dilation.


Subject(s)
CD36 Antigens/metabolism , Macrophages, Peritoneal/drug effects , Myocardial Infarction/drug therapy , Myocardium/metabolism , Phagocytosis/drug effects , Platelet Factor 4/toxicity , ADAMTS Proteins/metabolism , Animals , Cells, Cultured , Disease Models, Animal , Humans , Hypertrophy, Left Ventricular/chemically induced , Hypertrophy, Left Ventricular/metabolism , Hypertrophy, Left Ventricular/pathology , Hypertrophy, Left Ventricular/physiopathology , Infusions, Subcutaneous , Integrin beta4/metabolism , Macrophages, Peritoneal/metabolism , Macrophages, Peritoneal/pathology , Male , Matrix Metalloproteinase 9/metabolism , Mice, Inbred C57BL , Myocardial Infarction/metabolism , Myocardial Infarction/pathology , Myocardial Infarction/physiopathology , Myocardium/pathology , Platelet Factor 4/administration & dosage , Platelet Factor 4/metabolism , Signal Transduction , Ventricular Function, Left/drug effects , Ventricular Remodeling/drug effects
5.
Am J Physiol Heart Circ Physiol ; 315(3): H522-H530, 2018 09 01.
Article in English | MEDLINE | ID: mdl-29775405

ABSTRACT

The generation of big data has enabled systems-level dissections into the mechanisms of cardiovascular pathology. Integration of genetic, proteomic, and pathophysiological variables across platforms and laboratories fosters discoveries through multidisciplinary investigations and minimizes unnecessary redundancy in research efforts. The Mouse Heart Attack Research Tool (mHART) consolidates a large data set of over 10 yr of experiments from a single laboratory for cardiovascular investigators to generate novel hypotheses and identify new predictive markers of progressive left ventricular remodeling after myocardial infarction (MI) in mice. We designed the mHART REDCap database using our own data to integrate cardiovascular community participation. We generated physiological, biochemical, cellular, and proteomic outputs from plasma and left ventricles obtained from post-MI and no-MI (naïve) control groups. We included both male and female mice ranging in age from 3 to 36 mo old. After variable collection, data underwent quality assessment for data curation (e.g., eliminate technical errors, check for completeness, remove duplicates, and define terms). Currently, mHART 1.0 contains >888,000 data points and includes results from >2,100 unique mice. Database performance was tested, and an example is provided to illustrate database utility. This report explains how the first version of the mHART database was established and provides researchers with a standard framework to aid in the integration of their data into our database or in the development of a similar database. NEW & NOTEWORTHY The Mouse Heart Attack Research Tool combines >888,000 cardiovascular data points from >2,100 mice. We provide this large data set as a REDCap database to generate novel hypotheses and identify new predictive markers of adverse left ventricular remodeling following myocardial infarction in mice and provide examples of use. The Mouse Heart Attack Research Tool is the first database of this size that integrates data sets across platforms that include genomic, proteomic, histological, and physiological data.


Subject(s)
Databases, Factual , Myocardial Infarction/pathology , Software , Animals , Female , Male , Mice , Myocardial Infarction/physiopathology , Ventricular Remodeling
6.
JCI Insight ; 2(18)2017 09 21.
Article in English | MEDLINE | ID: mdl-28931761

ABSTRACT

Chronic inflammatory diseases, such as periodontal disease, associate with adverse wound healing in response to myocardial infarction (MI). The goal of this study was to elucidate the molecular basis for impaired cardiac wound healing in the setting of periodontal-induced chronic inflammation. Causal network analysis of 168 inflammatory and extracellular matrix genes revealed that chronic inflammation induced by a subseptic dose of Porphyromonas gingivalis lipopolysaccharide (LPS) exacerbated infarct expression of the proinflammatory cytokine Ccl12. Ccl12 prevented initiation of the reparative response by prolonging inflammation and inhibiting fibroblast conversion to myofibroblasts, resulting in diminished scar formation. Macrophage secretion of Ccl12 directly impaired fibronectin and collagen deposition and indirectly stimulated collagen degradation through upregulation of matrix metalloproteinase-2. In post-MI patients, circulating LPS levels strongly associated with the Ccl12 homologue monocyte chemotactic protein 1 (MCP-1). Patients with LPS levels ≥ 1 endotoxin units (EU)/ml (subseptic endotoxemia) at the time of hospitalization had increased end diastolic and systolic dimensions compared with post-MI patients with < 1 EU/ml, indicating that low yet pathological concentrations of circulating LPS adversely impact post-MI left ventricle (LV) remodeling by increasing MCP-1. Our study provides the first evidence to our knowledge that chronic inflammation inhibits reparative fibroblast activation and generates an unfavorable cardiac-healing environment through Ccl12-dependent mechanisms.


Subject(s)
Fibroblasts/metabolism , Macrophages/metabolism , Monocyte Chemoattractant Proteins/metabolism , Myocardium/metabolism , Periodontitis/metabolism , Wound Healing , Aged , Animals , Chronic Disease , Female , Humans , Lipopolysaccharides/administration & dosage , Male , Mice , Middle Aged , Myocardium/pathology , Periodontitis/chemically induced , Periodontitis/pathology , Systems Biology
7.
Am J Obstet Gynecol ; 217(4): 445.e1-445.e6, 2017 10.
Article in English | MEDLINE | ID: mdl-28599895

ABSTRACT

BACKGROUND: There are significant variations in gestational weight gain, with many women gaining in excess of the Institute of Medicine guidelines. Unfortunately, efforts to improve appropriate gestational weight gain have had only limited success. To date, interventions have focused primarily on decreasing energy intake and/or increasing physical activity. Maternal resting energy expenditure, which comprises ∼60% of total energy expenditure compared with the ∼20% that comes from physical activity, may be an important consideration in understanding variations in gestational weight gain. OBJECTIVE: Our objective was to quantify the changes in resting energy expenditure during pregnancy and their relationship to gestational weight gain and body composition changes among healthy women. We hypothesized that greater gestational weight gain, and fat mass accrual in particular, are inversely related to variations in resting energy expenditure. STUDY DESIGN: We conducted a secondary analysis of a prospective cohort studied before conception and late pregnancy (34-36 weeks). Body composition (estimated using hydrodensitometry) and resting energy expenditure (estimated using indirect calorimetry) were measured. The relationship between the changes in resting energy expenditure and gestational weight gain and the change in fat mass and fat-free mass were quantified. Resting energy expenditure was expressed as kilocalories per kilogram of fat-free mass per day (kilocalories per kilogram of fat-free mass-1/day-1) and kilocalories per day. Correlations are reported as r. RESULTS: Among 51 women, preconception body mass index was 23.0 (4.7) kg/m2; gestational weight gain was 12.8 (4.7) kg. Preconception and late pregnancy resting energy expenditure (kilocalories per day) correlated positively with the change in fat-free mass (r = 0.37, P = .008; r = 0.51, P = .001). Late-pregnancy resting energy expenditure (kilocalories per kilogram of fat-free mass-1/day-1) was inversely associated with the change in fat mass (r = -0.34, P = .02) and gestational weight gain (r = -0.29, P = .04). From before pregnancy through late gestation, the increase in resting energy expenditure (kilocalories per day) correlated positively with the change in fat-free mass (r = 0.44, P = .002) and negatively with the change in fat mass (r = -0.27, P = .06). CONCLUSION: The change in resting energy expenditure from before conception through late gestation correlated positively with changes in fat-free mass but negatively with fat mass accrual. Women with smaller increases in resting energy expenditure across pregnancy had greater gestational weight gain, specifically more adipose tissue. These data suggest that resting energy expenditure is an important factor in gestational weight gain, particularly excess fat mass accrual. Future lifestyle intervention studies need to consider clinically feasible means of estimating resting energy expenditure and, in response, tailor nutrient intake and composition recommendations. Implementing and testing such interventions would be a novel approach to improve compliance with gestational weight gain guidelines.


Subject(s)
Energy Metabolism , Rest , Weight Gain , Adult , Body Composition , Body Mass Index , Cohort Studies , Female , Humans , Pregnancy
8.
Am J Physiol Heart Circ Physiol ; 312(3): H375-H383, 2017 Mar 01.
Article in English | MEDLINE | ID: mdl-28011588

ABSTRACT

Advancing age is an independent risk factor for cardiovascular disease. Matrix metalloproteinase-9 (MMP-9) is secreted by macrophages and robustly increases in the left ventricle (LV) with age. The present study investigated the effect of MMP-9 overexpression in macrophages on cardiac aging. We compared 16- to 21-mo-old C57BL/6J wild-type (WT) and transgenic (TG) male and female mice (n = 15-20/group). MMP-9 overexpression amplified the hypertrophic response to aging, as evidenced by increased LV wall thickness and myocyte cross-sectional areas (P < 0.05 for both). MMP-9 overexpression reduced LV expression of the angiogenesis-related factors ICAM-1, integrins α3 and ß3, platelet/endothelial cell adhesion molecule-1, thrombospondin-1, tenascin-c, and versican (all P < 0.05). Concomitantly, the number of vessels in the TG was lower than WT LV (P < 0.05). This led to a mismatch in the muscle-to-vessel ratio and resulted in increased cardiac inflammation. Out of 84 inflammatory genes analyzed, 16 genes increased in the TG compared with WT (all P < 0.05). Of the elevated genes, 14 were proinflammatory genes. The increase in cardiac inflammation resulted in greater accumulation of interstitial collagen in TG (P < 0.05). Fractional shortening was similar between groups, indicating that global cardiac function was still preserved at this age. In conclusion, overexpression of MMP-9 in macrophages resulted in exacerbated cardiac hypertrophy in the setting of vessel rarefaction, which resulted in enhanced inflammation and fibrosis to augment the cardiac-aging phenotype. Our results provide evidence that macrophage-derived MMP-9 may be a therapeutic target in elderly subjects.NEW & NOTEWORTHY The present study was the first to use mice with transgenic overexpression of matrix metalloproteinase-9 (MMP-9) in macrophages to examine the effects of macrophage-derived MMP-9 on cardiac aging. We found that an elevation in macrophage-derived MMP-9 induced a greater age-dependent cardiac hypertrophy and vessel rarefaction phenotype, which enhanced cardiac inflammation and fibrosis.


Subject(s)
Aging/pathology , Blood Vessels/physiopathology , Cardiomegaly/genetics , Cardiomegaly/physiopathology , Inflammation/physiopathology , Matrix Metalloproteinase 9/genetics , Animals , Cardiomegaly/diagnostic imaging , Cell Size , Collagen/metabolism , Echocardiography , Female , Fibrosis , Heart Ventricles/pathology , Humans , Macrophages , Male , Mice , Mice, Inbred C57BL , Mice, Transgenic , Myocytes, Cardiac/pathology
9.
J Mol Cell Cardiol ; 100: 109-117, 2016 Nov.
Article in English | MEDLINE | ID: mdl-27746126

ABSTRACT

Matrix metalloproteinase-9 (MMP-9) is robustly elevated in the first week post-myocardial infarction (MI). Targeted deletion of the MMP-9 gene attenuates cardiac remodeling post-MI by reducing macrophage infiltration and collagen accumulation through increased apoptosis and reduced inflammation. In this study, we used a translational experimental design to determine whether selective MMP-9 inhibition early post-MI would be an effective therapeutic strategy in mice. We enrolled male C57BL/6J mice (3-6months old, n=116) for this study. Mice were subjected to coronary artery ligation. Saline or MMP-9 inhibitor (MMP-9i; 0.03µg/day) treatment was initiated at 3h post-MI and the mice were sacrificed at day (D) 1 or 7 post-MI. MMP-9i reduced MMP-9 activity by 31±1% at D1 post-MI (p<0.05 vs saline) and did not affect survival or infarct area. Surprisingly, MMP-9i treatment increased infarct wall thinning and worsened cardiac function at D7 post-MI. While MMP-9i enhanced neutrophil infiltration at D1 and macrophage infiltration at D7 post-MI, CD36 levels were lower in MMP-9i compared to saline, signifying reduced phagocytic potential per macrophage. Escalation and prolongation of the inflammatory response at D7 post-MI in the MMP-9i group was evident by increased expression of 18 pro-inflammatory cytokines (all p<0.05). MMP-9i reduced cleaved caspase 3 levels at D7 post-MI, consistent with reduced apoptosis and defective inflammation resolution. Because MMP-9i effects on inflammatory cells were significantly different from previously observed MMP-9 null mechanisms, we evaluated pre-MI (baseline) systemic differences between C57BL/6J and MMP-9 null plasma. By mass spectrometry, 34 plasma proteins were significantly different between groups, revealing a previously unappreciated altered baseline environment pre-MI when MMP-9 was deleted. In conclusion, early MMP-9 inhibition delayed inflammation resolution and exacerbated cardiac dysfunction, highlighting the importance of using translational approaches in mice.


Subject(s)
Matrix Metalloproteinase 9/metabolism , Myocardial Infarction/metabolism , Myocardial Infarction/physiopathology , Ventricular Dysfunction/metabolism , Animals , Apoptosis , Biomarkers , Cytokines/metabolism , Disease Models, Animal , Enzyme Activation , Extracellular Matrix/metabolism , Gene Expression , Immunohistochemistry , Inflammation/genetics , Inflammation/metabolism , Inflammation/pathology , Inflammation Mediators/metabolism , Macrophages/immunology , Macrophages/metabolism , Macrophages/pathology , Matrix Metalloproteinase 2/genetics , Matrix Metalloproteinase 2/metabolism , Matrix Metalloproteinase 9/genetics , Mice , Mortality , Myocardial Infarction/genetics , Myocardial Infarction/pathology , Neutrophil Infiltration , RNA Interference , Ventricular Dysfunction/genetics , Ventricular Dysfunction, Left/genetics , Ventricular Dysfunction, Left/metabolism , Ventricular Dysfunction, Left/pathology
10.
Am J Physiol Heart Circ Physiol ; 311(3): H822-36, 2016 09 01.
Article in English | MEDLINE | ID: mdl-27521418

ABSTRACT

The purpose of this study was to evaluate the effect of sham surgery in a minimally invasive surgical model of permanent coronary artery occlusion used to generate myocardial infarction (MI) in mice. Adult male C57BL/6J mice (3-6 mo old) were divided into five groups: day (D) 0 (no surgical operation), D1 Sham, D1 MI, D7 Sham, and D7 MI. A refined MI surgery technique was used to approach the coronary artery without the ribs being cut. Both sham and MI mice had the left ventricle (LV) exposed through a small incision. To test the effects of surgery alone, the suture was passed around the coronary artery but not ligated. The MI mice were subjected to permanent coronary artery ligation. The mice were killed at D1 or D7 postsurgical procedure. Compared with D0 no surgery controls, the D1 and D7 sham groups exhibited no surgical mortality and similar necropsy and echocardiographic variables. Surgery alone did not induce an inflammatory cell response, as evidenced by the lack of leukocyte infiltration in the sham groups. Analysis of 165 inflammatory cytokines and extracellular matrix factors in sham revealed that a minor gene response was initiated but not translated to protein levels. Collagen deposition did not occur in the absence of MI. In contrast, the D1 and D7 MI groups showed the expected robust inflammatory and scar formation responses. When a minimally invasive procedure to generate MI in mice was used, the D0 (no surgical operation) control was an adequate replacement for the use of sham surgery groups.


Subject(s)
Coronary Occlusion/metabolism , Coronary Vessels/surgery , Disease Models, Animal , Mice , Myocardial Infarction/metabolism , Myocardium/metabolism , Placebos , Animals , Collagen/metabolism , Coronary Occlusion/complications , Coronary Occlusion/pathology , Cytokines/metabolism , Extracellular Matrix/metabolism , Immunoblotting , Immunohistochemistry , Ligation , Male , Mice, Inbred C57BL , Minimally Invasive Surgical Procedures , Myocardial Infarction/etiology , Myocardial Infarction/pathology , Myocardium/pathology , Real-Time Polymerase Chain Reaction , Reverse Transcriptase Polymerase Chain Reaction
11.
Cardiovasc Res ; 110(1): 51-61, 2016 May 01.
Article in English | MEDLINE | ID: mdl-26825554

ABSTRACT

AIMS: Although macrophage phenotypes have been well studied in the myocardial infarction (MI) setting, this study investigated temporal neutrophil polarization and activation mechanisms. METHODS AND RESULTS: Neutrophils isolated from the infarcted left ventricle (LV) of mice showed high expression of proinflammatory markers at Day 1 and anti-inflammatory markers at Days 5 and 7 post-MI, indicating distinct neutrophil phenotypes along the post-MI time continuum. Flow cytometry analysis revealed that although proinflammatory N1 neutrophils were always predominant (>80% of total neutrophils at each time point), the percentage of N2 neutrophils increased post-MI from 2.4 ± 0.6% at Day 1 to 18.1 ± 3.0% at Day 7. In vitro, peripheral blood neutrophils were polarized to proinflammatory N1 by lipopolysaccharide and interferon-γ or anti-inflammatory N2 by interleukin-4, indicating high plasticity potential. The in vivo post-MI relevant LV damage-associated molecular patterns (DAMPs) polarized neutrophils to a proinflammatory N1 phenotype by activating toll-like receptor-4. Transforming growth factor-ß1 inhibited proinflammatory production in neutrophils. N1 neutrophils positively correlated with infarct wall thinning at Day 7 post-MI, possibly due to high production of matrix metalloproteinases-12 and -25. CONCLUSION: This study is the first to identify the existence of N1 and N2 neutrophils in the infarct region and reveals that N1 polarization could be mediated by DAMPs.


Subject(s)
Macrophages/metabolism , Myocardial Infarction/metabolism , Neutrophils/metabolism , Ventricular Remodeling/physiology , Animals , Cell Polarity , Disease Models, Animal , Inflammation/metabolism , Interferon-gamma/metabolism , Interleukin-4/metabolism , Male , Mice, Inbred C57BL , Myocardial Infarction/pathology
12.
J Am Coll Cardiol ; 66(12): 1364-74, 2015 Sep 22.
Article in English | MEDLINE | ID: mdl-26383724

ABSTRACT

BACKGROUND: Proteolytically released extracellular matrix (ECM) fragments, matricryptins, are biologically active and play important roles in wound healing. Following myocardial infarction (MI), collagen I, a major component of cardiac ECM, is cleaved by matrix metalloproteinases (MMPs). OBJECTIVES: This study identified novel collagen-derived matricryptins generated post-MI that mediate remodeling of the left ventricle (LV). METHODS: Recombinant collagen Ia1 was used in MMPs cleavage assays, the products were analyzed by mass spectrometry for identification of cleavage sites. C57BL6/J mice were given MI and animals were treated either with vehicle control or p1158/59 matricryptin. Seven days post-MI, LV function and parameters of LV remodeling were measured. Levels of p1158/59 were also measured in plasma of MI patients and healthy controls. RESULTS: In situ, MMP-2 and -9 generate a collagen Iα1 C-1158/59 fragment, and MMP-9 can further degrade it. The C-1158/59 fragment was identified post-MI, both in human plasma and mouse LV, at levels that inversely correlated to MMP-9 levels. We synthesized a peptide beginning at the cleavage site (p1158/59, amino acids 1159 to 1173) to investigate its biological functions. In vitro, p1158/59 stimulated fibroblast wound healing and robustly promoted angiogenesis. In vivo, early post-MI treatment with p1158/59 reduced LV dilation at day 7 post-MI by preserving LV structure (p < 0.05 vs. control). The p1158/59 stimulated both in vitro and in vivo wound healing by enhancing basement membrane proteins, granulation tissue components, and angiogenic factors. CONCLUSIONS: Collagen Iα1 matricryptin p1158/59 facilitates LV remodeling post-MI by regulating scar formation through targeted ECM generation and stimulation of angiogenesis.


Subject(s)
Collagen Type I/metabolism , Collagen Type I/therapeutic use , Myocardial Infarction/metabolism , Ventricular Remodeling , Amino Acid Sequence , Animals , Cicatrix , Collagen Type I/blood , Collagen Type I/pharmacology , Collagen Type I, alpha 1 Chain , Drug Evaluation, Preclinical , Extracellular Matrix/metabolism , Female , Human Umbilical Vein Endothelial Cells , Humans , Male , Matrix Metalloproteinase 9/metabolism , Mice, Inbred C57BL , Middle Aged , Molecular Sequence Data , Neovascularization, Physiologic , Random Allocation , Wound Healing
13.
Crit Care Med ; 43(10): 2049-2058, 2015 Oct.
Article in English | MEDLINE | ID: mdl-26086942

ABSTRACT

OBJECTIVE: Sepsis remains a predominant cause of mortality in the ICU, yet strategies to increase survival have proved largely unsuccessful. This study aimed to identify proteins linked to sepsis outcomes using a glycoproteomic approach to target extracellular proteins that trigger downstream pathways and direct patient outcomes. DESIGN: Plasma was obtained from the Lactate Assessment in the Treatment of Early Sepsis cohort. N-linked plasma glycopeptides were quantified by solid-phase extraction coupled with mass spectrometry. Glycopeptides were assigned to proteins using RefSeq (National Center of Biotechnology Information, Bethesda, MD) and visualized in a heat map. Protein differences were validated by immunoblotting, and proteins were mapped for biological processes using Database for Annotation, Visualization and Integrated Discovery (National Institute of Allergy and Infectious Diseases, National Institutes of Health; Bethesda, MD) and for functional pathways using Kyoto Encyclopedia of Genes and Genomes (Kanehisa Laboratories, Kyoto, Japan) databases. SETTING: Hospitalized care. PATIENTS: Patients admitted to the emergency department were enrolled in the study when the diagnosis of sepsis was made, within 6 hours of presentation. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: A total of 501 glycopeptides corresponding to 234 proteins were identified. Of these, 66 glycopeptides were unique to the survivor group and corresponded to 54 proteins, 60 were unique to the nonsurvivor group and corresponded to 43 proteins, and 375 were common responses between groups and corresponded to 137 proteins. Immunoblotting showed that nonsurvivors had increased total kininogen; decreased total cathepsin-L1, vascular cell adhesion molecule, periostin, and neutrophil gelatinase-associated lipocalin; and a two-fold decrease in glycosylated clusterin (all p < 0.05). Kyoto Encyclopedia of Genes and Genomes analysis identified six enriched pathways. Interestingly, survivors relied on the extrinsic pathway of the complement and coagulation cascade, whereas nonsurvivors relied on the intrinsic pathway. CONCLUSION: This study identifies proteins linked to patient outcomes and provides insight into unexplored mechanisms that can be investigated for the identification of novel therapeutic targets.


Subject(s)
Glycoproteins/blood , Proteomics , Sepsis/blood , Sepsis/mortality , Aged , Female , Humans , Male , Predictive Value of Tests
14.
J Matern Fetal Neonatal Med ; 28(10): 1113-5, 2015 Jul.
Article in English | MEDLINE | ID: mdl-25012807

ABSTRACT

CONTEXT: There are many causes of impaired glucose tolerance in pregnant women. It is unclear whether genetic etiologies are a source of impaired glucose tolerance in pregnant women. OBJECTIVE: To prospectively determine the prevalence of maturity onset diabetes of the young (MODY) due to glucokinase (GCK) mutations in an American population of women with recent onset diabetes mellitus and gestational diabetes. We hypothesized that based on America's higher prevalence of gestational diabetes mellitus (GDM) and Type 2 diabetes, there may be an increased prevalence of GK mutations in our population than in previously reported studies from European studies. DESIGN: Over a three-year period, 72 pregnant women with recently diagnosed diabetes mellitus were prospectively assessed for presence of the most common pathogenic GCK mutations. SETTING: This study was performed in a gestational diabetes clinic in Urban America and a high-risk pregnancy clinic that served the military and their families on an American military base in Germany. PATIENTS: Seventy-two women; 65 with diagnosis of diabetes mellitus in this pregnancy (GDM/overt diabetes) and 7 with diagnosis in the last nine years prior to pregnancy were recruited during pregnancy and blood samples were obtained. INTERVENTIONS: None. MAIN OUTCOME MEASURES: Each study participant's blood sample was analyzed with restriction fragment length polymorphism to assess for mutations in the GCK gene. RESULTS: There were 38 female and 34 male neonates born at 38 weeks gestation ± 1.2 weeks. Mean birth weight was 3351 g ± 450 g. There were no patients with GCK mutations found in our population 0/72. This prevalence is not greater than that seen in previous a similar study in European women with gestational diabetes, but in fact significantly less (p = 0.03). CONCLUSION: American women with recently diagnosed diabetes mellitus likely have no higher prevalence of MODY than in previously studied European women with diabetes mellitus and may have a lower prevalence.


Subject(s)
Diabetes Mellitus, Type 2/epidemiology , Diabetes, Gestational/genetics , Glucokinase/genetics , Mutation , Adult , Female , Germany/epidemiology , Humans , Male , Polymorphism, Restriction Fragment Length , Pregnancy , Pregnancy, High-Risk , Prevalence , Prospective Studies , United States/epidemiology , White People/genetics , Young Adult
15.
Pediatr Obes ; 9(6): e141-4, 2014 Dec.
Article in English | MEDLINE | ID: mdl-25267097

ABSTRACT

BACKGROUND: Rapid weight gain in infancy has been established as a risk factor for the development of later obesity. OBJECTIVE: We aimed to investigate the role of changes in infant body composition (assessed via total body electrical conductivity) on the development of overweight/obesity in mid-childhood. METHODS: Fifty-three term infants were evaluated at birth, three times during infancy and in mid-childhood. Logistic regression was used to determine associations between rates of total weight gain, fat mass gain and lean mass gain during infancy and later overweight/obesity (defined as body mass index [BMI] ≥85th percentile), adjusted for birth weight and parent education. RESULTS: At follow-up (age 9.0 ± 1.8 years), 30% were overweight/obese. More rapid total weight gain from 0 to 4 months was associated with twofold odds (odds ratio [OR] 1.98, 95% confidence interval [CI] 1.05-3.74, P = 0.04) of overweight/obesity in mid-childhood. From 0 to 8 months, more rapid weight gain was associated with nearly fivefold odds (OR 4.76, 95% CI 1.05-21.5, P = 0.04), and more rapid fat mass gain was associated with eightfold odds (OR 8.03, 95% CI 1.11-58.2, P = 0.04) of later overweight/obesity. CONCLUSION: This exploratory study suggests that rapid weight gain, especially fat mass gain, in earlier infancy predisposes to mid-childhood overweight/obesity.


Subject(s)
Body Composition , Pediatric Obesity/epidemiology , Weight Gain , Birth Weight , Body Mass Index , Causality , Child , Child, Preschool , Female , Follow-Up Studies , Humans , Infant , Infant, Newborn , Logistic Models , Longitudinal Studies , Male , Mothers , Odds Ratio , Pediatric Obesity/prevention & control , Predictive Value of Tests , Pregnancy , Prospective Studies , Risk Factors
16.
J Perinatol ; 33(12): 934-8, 2013 Dec.
Article in English | MEDLINE | ID: mdl-24030674

ABSTRACT

OBJECTIVE: To examine changes in skinfolds in late gestation in healthy women. STUDY DESIGN: Skinfold measures were performed in 39 women at 30.8 (mean) and 37.7 weeks gestation. Fat mass (kg) and sum of three skinfolds were calculated. RESULTS: A decrease in skinfold thickness was observed in 21 women (-3.1±2.1 mm) in late gestation, whereas 18 women had an increase (4.3±3.2 mm), P<0.001. The group of women who lost body fat (decrease in skinfold thickness) had a trend toward greater pregravid body mass index (BMI, 25 vs 22 kg/m(2), P=0.06), and gained less weight in late gestation (3.0 vs 4.3 kg, P=0.042). On multiple regression, maternal age and gestational weight gain were positively correlated with fat mass accrual, whereas pregravid BMI and dietary fiber were negative determinants of late gestational fat mass. CONCLUSION: Increases in maternal fat mass in late gestation were related to maternal age and gestational weight gain, whereas decreases were related to increased pregravid BMI and dietary fiber.


Subject(s)
Adipose Tissue/physiology , Pregnancy Trimester, Third/physiology , Skinfold Thickness , Adult , Body Mass Index , Dietary Fiber , Female , Humans , Maternal Age , Pregnancy , Regression Analysis , Weight Gain
17.
J Clin Endocrinol Metab ; 93(7): 2885-90, 2008 Jul.
Article in English | MEDLINE | ID: mdl-18445668

ABSTRACT

CONTEXT: High plasma adiponectin concentrations in human fetuses and neonates are unique features of early developmental stages. Yet, the origins of the high adiponectin concentrations in the perinatal period remain elusive. OBJECTIVE: This study was undertaken to identify the sources and functional properties of adiponectin in utero. DESIGN AND METHODS: Tissue specimens were obtained at autopsy from 21- to 39-wk-old stillborn human fetuses. Adipose tissue and placenta were obtained at term elective cesarean section. Adiponectin complexes and expression were measured by immunodetection and real-time PCR. RESULTS: Adiponectin mRNA transcripts were detected in fetal sc and omental adipose depots at lower concentrations than in maternal adipose tissue. Immunoreactive adiponectin was also observed in vascular endothelial cells of fetal organs, including skeletal muscle, kidney, and brain. The absence of adiponectin in all placental cell types and lack of correlation between maternal and umbilical adiponectin indicate that umbilical adiponectin reflects its exclusive production by fetal tissues. The most prominent forms of adiponectin in fetal plasma were high and low molecular mass (HMW and LMW) multimers of 340 and 160 kDa, respectively. The proportion of the HMW complexes was 5-fold (P < 0.001) higher in umbilical plasma than in adult. The high HMW and total adiponectin levels were associated with lower insulin concentration and lower homeostasis model of assessment of insulin resistance indices in umbilical plasma, reflecting higher insulin sensitivity of the fetus compared with adult. CONCLUSIONS: The abundance of HMW adiponectin and its vascular expression are characteristics of human fetal adiponectin. Combined with high insulin sensitivity, fetal adiponectin may be a critical determinant of in utero growth.


Subject(s)
Adiponectin/analysis , Endothelial Cells/chemistry , Fetus/chemistry , Adiponectin/genetics , Fetus/metabolism , Humans , Infant, Newborn , Insulin Resistance , Molecular Weight , RNA, Messenger/analysis
18.
Diabetologia ; 49(7): 1677-85, 2006 Jul.
Article in English | MEDLINE | ID: mdl-16752186

ABSTRACT

AIMS/HYPOTHESIS: Adiponectin is upregulated during adipogenesis and downregulated in insulin-resistant states. The mechanism(s) governing the re-arrangements from adipogenesis to facilitated lipolysis during pregnancy are unknown. Our purpose was to analyse the role of adiponectin relative to the metabolic changes in human pregnancy. SUBJECTS, MATERIALS AND METHODS: Lean women (BMI <25 kg/m(2)) were evaluated longitudinally before conception, and in early (12-14 weeks) and late (34-36 weeks) pregnancy. Insulin sensitivity was measured using the glucose clamp technique. Venous blood and subcutaneous adipose tissue biopsies were obtained at each time point. RESULTS: Adiponectin concentrations were lower in the third trimester than in the pregravid condition (9.9+/-1.4 vs 13.5+/-1.8 microg/ml). The hypoadiponectinaemia was reflected by a 2.5-fold decrease in white adipose tissue adiponectin mRNA. These changes were associated with a 25% increase in fat mass (23.7+/-2.9 vs 18.9+/-2.9 kg). Insulin infusion decreased high molecular weight adiponectin complexes in pregravid women (9.9+/-0.6 vs 6.2+/-0.06) and the suppressive effect of insulin was lost during pregnancy. The pregnancy-mediated changes in adiponectin were strongly correlated with basal insulin levels and insulin sensitivity (p<0.0001). The relationship between adiponectin and insulin sensitivity was related to the decreased insulin regulation of glucose utilisation (r=0.55, p<0.001) but not of endogenous hepatic glucose production. CONCLUSIONS/INTERPRETATION: These data demonstrate that pregnancy is associated with adiponectin changes in lean women. Hypoadiponectinaemia is reflected by a lower amount of high molecular weight adiponectin and by the ratio of high to low molecular weight multimers. The adiponectin changes relate to decreased insulin sensitivity of glucose disposal rather than alterations of lipid metabolism.


Subject(s)
Adiponectin/physiology , Glucose/metabolism , Lipid Metabolism , Pregnancy/metabolism , Adiponectin/blood , Blood Glucose/analysis , Energy Metabolism , Female , Gestational Age , Glucose Clamp Technique , Humans , Insulin/blood , Insulin/pharmacology , Pregnancy/blood
19.
J Matern Fetal Neonatal Med ; 12(6): 396-401, 2002 Dec.
Article in English | MEDLINE | ID: mdl-12683650

ABSTRACT

OBJECTIVE: To determine whether neonatal fat mass, which may be a better estimate of fetal overgrowth, is correlated with maternal fasting, preprandial and/or postprandial glucose values in women with gestational diabetes mellitus (GDM). STUDY DESIGN: Women with GDM and no other medical or obstetric problems, and their infants, were the subjects of this study. Portable reflectance meters were used by all participants for self-monitoring of blood glucose levels. Average fasting, preprandial, 2-h postprandial and bedtime glucose values were determined for each subject. Neonatal body composition was obtained by total body electric conductivity and/or anthropometric measurements within 48 h after delivery. RESULTS: Eighteen women with their infants participated in this study. The age (mean +/- SD) of the mothers was 28.0 +/- 5.7 years. Nine were treated with diet and nine with diet and insulin. An average of 40 fasting (84 +/- 13 mg/dl), 50 preprandial (87 +/- 14 mg/dl), 80 2-h postprandial (106 +/- 19 mg/dl) and 17 bedtime (104 +/- 19 mg/dl) glucose values were obtained from each subject. The average gestational age of the infants at birth was 38.3 +/- 1.3 weeks with a mean weight of 3,356 +/- 526 g. Three infants were > 4 kg and seven infants were > 90th centile for gestational age. The strongest correlation with neonatal fat mass was maternal fasting glucose level (r = 0.71, p < 0.01). Neonatal fat mass was not found to be significantly correlated with any other mean glucose value. Additionally, the infant's per cent body fat (r = 0.71, p < 0.01), sum of skinfold thicknesses (r = 0.70, p < 0.01), fat-free mass (r = 0.50, p < 0.05), and weight (r = 0.61, p < 0.01) were also found to be correlated with maternal fasting glucose level. No other maternal glucose measurements were correlated with either birth weight or estimates of fat free mass. CONCLUSION: Maternal fasting glucose levels correlated best with neonatal fat mass and other estimates of neonatal body composition.


Subject(s)
Birth Weight/physiology , Blood Glucose Self-Monitoring , Body Composition/physiology , Diabetes, Gestational/blood , Infant, Newborn/growth & development , Adult , Diabetes, Gestational/physiopathology , Diabetes, Gestational/therapy , Diet, Diabetic , Fasting , Female , Humans , Hypoglycemic Agents/therapeutic use , Insulin/therapeutic use , Linear Models , Postprandial Period , Pregnancy , Pregnancy Outcome , Regression Analysis , Time Factors
20.
Am J Obstet Gynecol ; 185(6): 1416-20, 2001 Dec.
Article in English | MEDLINE | ID: mdl-11744918

ABSTRACT

OBJECTIVE: This study seeks to determine which parental demographic and metabolic factors best correlate with fetal growth and body composition as estimated by ultrasound. STUDY DESIGN: Thirty-one gravid patients had ultrasound estimates of fetal anthropometry in mid-third trimester. These measurements included estimated fetal weight, abdominal subcutaneous fat, and/or thigh subcutaneous fat thickness. Independent variables included diagnosis of gestational diabetes, parental demographic factors, neonatal sex, and late gestation estimates of carbohydrate metabolism. RESULTS: In the multivariate stepwise model the strongest predictor of ultrasound estimated fetal weight was basal hepatic glucose production, followed by late gestation insulin sensitivity (total R (2) = 0.27). The strongest predictors of abdominal subcutaneous fat thickness were weight gain and presence of gestational diabetes (total R (2) = 0.25). CONCLUSION: Measures of maternal carbohydrate metabolism, rather than fat mass, explain sonographic measurements of fetal weight. We speculate that factors other than maternal carbohydrate metabolism further explain the variances of fetal adiposity.


Subject(s)
Body Composition , Embryonic and Fetal Development , Pregnancy/metabolism , Ultrasonography, Prenatal , Abdomen/diagnostic imaging , Abdomen/embryology , Adipose Tissue/diagnostic imaging , Adipose Tissue/embryology , Adult , Diabetes, Gestational/diagnostic imaging , Female , Fetal Weight , Forecasting , Gestational Age , Glucose/biosynthesis , Humans , Insulin/physiology , Liver/metabolism , Weight Gain
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