65 results on '"Wijesinghe DS"'
Search Results
2. Active Open Market Operations: A Review of Experience
- Author
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Wijesinghe, DS, primary
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- 2009
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3. Microvertebrate faunal assemblages of the Favel Formation (late Cenomanian-middle Turonian) of Manitoba, Canada.
- Author
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Kilmury AA, Anderson A, Wijesinghe DS, Verstraete AF, Ezeana W, Anderson AE, and Brink KS
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- Animals, Manitoba, Reptiles, Canada, Fossils, Vertebrates
- Abstract
Microvertebrate assemblages of the Upper Cretaceous (late Cenomanian to mid-Turonian) Favel Formation of Manitoba are formally described for the first time. New vertebrate occurrences from the Favel Formation include the actinopterygians Caturidae indet., cf. Albulidae incertae sedis , Micropycnodon kansasensis , Pachyrhizodus minimus , Protosphyraena sp., Thryptodus loomisi , chondrichthyans Ischyrhiza cf. mira , I. texana , Ptychodus marginalis , P. occidentalis , and P. rhombodus , the avian cf. Ichthyornis sp., the reptile Testudines indet., and an unknown taxon referred to as Vertebrate A. Changes in faunal occurrences throughout the formation suggest an offshore open marine environment for the lower and middle horizons and nearshore marine for the upper horizon, represent ing mid- and late stages of the Greenhorn third-order marine cycle. This newly described diversity increases biogeographic affinities of the late Cenomanian to mid-Turonian vertebrate assemblages of Manitoba with central WIS localities in South Dakota and Kansas, providing additional support for a central vertebrate biogeographic subprovince during late Cenomanian to early Turonian times, as well as WIS localities further south in Texas decreasing the gradient of the north-south or central-south community boundary during early and mid-Turonian times., Competing Interests: The authors declare that they have no competing interests., (© 2023 Kilmury et al.)
- Published
- 2023
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4. Pseudomonas Aeruginosa Theft Biofilm Require Host Lipids of Cutaneous Wound.
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Sinha M, Ghosh N, Wijesinghe DS, Mathew-Steiner SS, Das A, Singh K, El Masry M, Khanna S, Inoue H, Yamazaki K, Kawada M, Gordillo GM, Roy S, and Sen CK
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- Animals, Ceramidases, Lower Extremity, Swine, PPAR delta, Pseudomonas aeruginosa
- Abstract
Objective: This work addressing complexities in wound infection, seeks to test the reliance of bacterial pathogen Pseudomonas aeruginosa (PA) on host skin lipids to form biofilm with pathological consequences., Background: PA biofilm causes wound chronicity. Both CDC as well as NIH recognizes biofilm infection as a threat leading to wound chronicity. Chronic wounds on lower extremities often lead to surgical limb amputation., Methods: An established preclinical porcine chronic wound biofilm model, infected with PA or Pseudomonas aeruginosa ceramidase mutant (PA ∆Cer ), was used., Results: We observed that bacteria drew resource from host lipids to induce PA ceramidase expression by three orders of magnitude. PA utilized product of host ceramide catabolism to augment transcription of PA ceramidase. Biofilm formation was more robust in PA compared to PA ∆Cer . Downstream products of such metabolism such as sphingosine and sphingosine-1-phosphate were both directly implicated in the induction of ceramidase and inhibition of peroxisome proliferator-activated receptor (PPAR)δ, respectively. PA biofilm, in a ceram-idastin-sensitive manner, also silenced PPARδ via induction of miR-106b. Low PPARδ limited ABCA12 expression resulting in disruption of skin lipid homeostasis. Barrier function of the wound-site was thus compromised., Conclusions: This work demonstrates that microbial pathogens must co-opt host skin lipids to unleash biofilm pathogenicity. Anti-biofilm strategies must not necessarily always target the microbe and targeting host lipids at risk of infection could be productive. This work may be viewed as a first step, laying fundamental mechanistic groundwork, toward a paradigm change in biofilm management., Competing Interests: The authors report no conflict of interest., (Copyright © 2021 The Author(s). Published by Wolters Kluwer Health, Inc.)
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- 2023
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5. Histone acetylation at the sulfotransferase 1a1 gene is associated with its hepatic expression in normal aging.
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Kronfol MM, Abudahab S, Dozmorov MG, Jahr FM, Halquist MS, McRae M, Wijesinghe DS, Price ET, Slattum PW, and McClay JL
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- Acetylation, Aged, Aging genetics, Animals, Humans, Liver metabolism, Mice, Sulfotransferases genetics, Sulfotransferases metabolism, Epigenesis, Genetic, Histones genetics, Histones metabolism
- Abstract
Objectives: Phase II drug metabolism is poorly studied in advanced age and older adults may exhibit significant variability in their expression of phase II enzymes. We hypothesized that age-related changes to epigenetic regulation of genes involved in phase II drug metabolism may contribute to these effects., Methods: We examined published epigenome-wide studies of human blood and identified the SULT1A1 and UGT1A6 genes as the top loci showing epigenetic changes with age. To assess possible functional alterations with age in the liver, we assayed DNA methylation (5mC) and histone acetylation changes around the mouse homologs Sult1a1 and Ugt1a6 in liver tissue from mice aged 4-32 months., Results: Our sample shows a significant loss of 5mC at Sult1a1 (β = -1.08, 95% CI [-1.8, -0.2], SE = 0.38, P = 0.011), mirroring the loss of 5mC with age observed in human blood DNA at the same locus. We also detected increased histone 3 lysine 9 acetylation (H3K9ac) with age at Sult1a1 (β = 0.11, 95% CI [0.002, 0.22], SE = 0.05, P = 0.04), but no change to histone 3 lysine 27 acetylation (H3K27ac). Sult1a1 gene expression is significantly positively associated with H3K9ac levels, accounting for 23% of the variation in expression. We did not detect any significant effects at Ugt1a6., Conclusions: Sult1a1 expression is under epigenetic influence in normal aging and this influence is more pronounced for H3K9ac than DNA methylation or H3K27ac in this study. More generally, our findings support the relevance of epigenetics in regulating key drug-metabolizing pathways. In the future, epigenetic biomarkers could prove useful to inform dosing in older adults., (Copyright © 2021 Wolters Kluwer Health, Inc. All rights reserved.)
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- 2021
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6. Staphylococcus aureus Lipase 3 (SAL3) is a surface-associated lipase that hydrolyzes short chain fatty acids.
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Kumar NG, Contaifer D Jr, Wijesinghe DS, and Jefferson KK
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- Genome, Bacterial genetics, Host-Pathogen Interactions genetics, Humans, Hydrolysis, Staphylococcal Infections enzymology, Staphylococcus aureus genetics, Staphylococcus aureus pathogenicity, Lipase genetics, Lipids genetics, Staphylococcal Infections microbiology, Staphylococcus aureus enzymology
- Abstract
Bacterial lipases play important roles during infection. The Staphylococcus aureus genome contains several genes that encode well-characterized lipases and several genes predicted to encode lipases or esterases for which the function has not yet been established. In this study, we sought to define the function of an uncharacterized S. aureus protein, and we propose the annotation S. aureus lipase 3 (SAL3) (SAUSA300_0641). We confirmed that SAL3 is a lipase and that it is surface associated and secreted through an unknown mechanism. We determined that SAL3 specifically hydrolyzes short chain (4-carbon and fewer) fatty acids and specifically binds negatively charged lipids including phosphatidic acid, phosphatidylinositol phosphate, and phosphatidylglycerol, which is the most abundant lipid in the staphylococcal cell membrane. Mutating the catalytic triad S66-A, D167-A, S168-A, and H301-A in the recombinant protein abolished lipase activity without altering binding to host lipid substrates. Taken together we report the discovery of a novel lipase from S. aureus specific to short chain fatty acids with yet to be determined roles in host pathogen interactions., Competing Interests: The authors have declared that no competing interests exist.
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- 2021
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7. Using Literature Based Discovery to Gain Insights Into the Metabolomic Processes of Cardiac Arrest.
- Author
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Henry S, Wijesinghe DS, Myers A, and McInnes BT
- Abstract
In this paper, we describe how we applied LBD techniques to discover lecithin cholesterol acyltransferase (LCAT) as a druggable target for cardiac arrest. We fully describe our process which includes the use of high-throughput metabolomic analysis to identify metabolites significantly related to cardiac arrest, and how we used LBD to gain insights into how these metabolites relate to cardiac arrest. These insights lead to our proposal (for the first time) of LCAT as a druggable target; the effects of which are supported by in vivo studies which were brought forth by this work. Metabolites are the end product of many biochemical pathways within the human body. Observed changes in metabolite levels are indicative of changes in these pathways, and provide valuable insights toward the cause, progression, and treatment of diseases. Following cardiac arrest, we observed changes in metabolite levels pre- and post-resuscitation. We used LBD to help discover diseases implicitly linked via these metabolites of interest. Results of LBD indicated a strong link between Fish Eye disease and cardiac arrest. Since fish eye disease is characterized by an LCAT deficiency, it began an investigation into the effects of LCAT and cardiac arrest survival. In the investigation, we found that decreased LCAT activity may increase cardiac arrest survival rates by increasing ω -3 polyunsaturated fatty acid availability in circulation. We verified the effects of ω -3 polyunsaturated fatty acids on increasing survival rate following cardiac arrest via in vivo with rat models., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Henry, Wijesinghe, Myers and McInnes.)
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- 2021
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8. Effects of ω-3 PUFA and ascorbic acid combination on post-resuscitation myocardial function.
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Cheng C, Li H, Liang L, Jin T, Zhang G, Bradley JL, Peberdy MA, Ornato JP, Wijesinghe DS, and Tang W
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- Animals, Biomarkers blood, Disease Models, Animal, Heart Arrest blood, Heart Arrest physiopathology, Inflammation Mediators blood, Lipid Peroxidation drug effects, Male, Oxidative Stress drug effects, Rats, Sprague-Dawley, Recovery of Function, Ventricular Fibrillation blood, Ventricular Fibrillation physiopathology, Rats, Anti-Inflammatory Agents pharmacology, Antioxidants pharmacology, Ascorbic Acid pharmacology, Blood Circulation drug effects, Cardiopulmonary Resuscitation, Fatty Acids, Omega-3 pharmacology, Heart Arrest therapy, Myocardium metabolism, Ventricular Fibrillation therapy
- Abstract
Accumulating evidence demonstrated that administration of ω-3 polyunsaturated fatty acid (ω-3 PUFA) or ascorbic acid (AA) following cardiac arrest (CA) improves survival. Therefore, we investigate the effects of ω-3 PUFA combined with AA on myocardial function after CA and cardiopulmonary resuscitation (CPR) in a rat model. Thirty male rats were randomized into 5 groups: (1) sham; (2) control; (3) ω-3 PUFA; (4) AA; (5) ω-3 PUFA + AA. Ventricular fibrillation (VF) was induced and untreated for 6 min followed by defibrillation after 8 min of CPR. Infusion of drug or vehicle occurred at the start of CPR. Myocardial function and sublingual microcirculation were measured at baseline and after return of spontaneous circulation (ROSC). Heart tissues and blood were collected 6 h after ROSC. Myocardial function and sublingual microcirculation improvements were seen with ω-3 PUFA or AA compared to control after ROSC (p < 0.05). ω-3 PUFA + AA shows a better myocardial function than ω-3 PUFA or AA (p < 0.05). ω-3 PUFA or AA decreases pro-inflammatory cytokines, cTnI, myocardium malondialdehyde (MDA) and 4-hydroxynonenal (4-HNE) modified proteins compared to control (p < 0.05). ω-3 PUFA and AA combined have lower MDA and 4-HNE modified proteins than alone (p < 0.05). ω-3 PUFA or AA treatment reduces the severity of post-resuscitation myocardial dysfunction, improves sublingual microcirculation, decreases lipid peroxidation and systemic inflammation in the early phase of recovery following CA and resuscitation. A combination of ω-3 PUFA and AA treatment confers an additive effect in suppressing lipid peroxidation and improving myocardial function., (Copyright © 2020 The Authors. Published by Elsevier Masson SAS.. All rights reserved.)
- Published
- 2021
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9. Cannabinoid Receptor Agonist WIN55, 212-2 Adjusts Lipid Metabolism in a Rat Model of Cardiac Arrest.
- Author
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Xiao Y, Contaifer D, Huang W, Yang J, Hu Z, Guo Q, Bradley J, Peberdy MA, Ornato JP, Wijesinghe DS, and Tang W
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- Animals, Cannabinoid Receptor Agonists pharmacology, Disease Models, Animal, Lipid Metabolism, Rats, Rats, Sprague-Dawley, Ventricular Fibrillation therapy, Cardiopulmonary Resuscitation, Heart Arrest therapy, Hypothermia, Induced
- Abstract
The objective of this study was to investigate the effects of pharmacologically induced hypothermia with WIN55, 212-2 (WIN)on postresuscitation myocardial function, microcirculation, and metabolism-specific lipids in a rat cardiac arrest (CA) model. Ventricular fibrillation was electrically induced and untreated for 6 minutes in 24 Sprague-Dawley rats weighing 450-550 g. Cardiopulmonary resuscitation including chest compression and mechanical ventilation was then initiated and continued for 8 minutes, followed by defibrillation. At 5 minutes after restoration of spontaneous circulation (ROSC), animals were randomized into four groups: (1) normothermia with vehicle (NT); (2) physical hypothermia with vehicle (PH); (3) WIN55, 212-2 with normothermia (WN); and (4) WIN55, 212-2 with hypothermia (WH). For groups of WN and WH, WIN was administered by continuous intravenous infusion with a syringe pump for 4 hours. PH started at 5 minutes after resuscitation. NT maintained core temperature at 37°C ± 0.2°C with the aid of a heating blanket. Hypothermia groups maintained temperature at 33°C ± 0.5°C for 4 hours after ROSC. There was a significant improvement in myocardial function as measured by ejection fraction, cardiac output, and myocardial performance index in animals treated with WH and PH beginning at 1 hour after start of infusion. In the WH and PH groups, buccal microcirculation was significantly improved compared with NT and WN. Plasma at pre-CA and ROSC 4 hours was harvested for lipid metabolism. The WH group appeared to be closer to baseline than the other groups in lipid metabolism. lysophosphatidylcholine (LPC) 18:2, free fatty acid (FFA) 22:6, and ceramide (CER) (24:0) changed significantly among the lipidomic data compared with NT ( p < 0.05). Postresuscitation hypothermia improved myocardial function and microcirculation. WH-mediated lipid metabolism had the best metabolic outcome to bring back the animals to normal metabolism, which may be protective to improve outcomes of CA. LPC 18:2, FFA 22:6, and CER (24:0) may be important predictors of outcomes of CA.
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- 2020
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10. Pulmonary delivery of ORMDL3 short hairpin RNA - a potential tool to regulate allergen-induced airway inflammation.
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Dileepan M, Ha SG, Rastle-Simpson S, Ge XN, Greenberg YG, Wijesinghe DS, Contaifer D Jr, Rao SP, and Sriramarao P
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- Animals, Asthma drug therapy, Asthma metabolism, Bronchial Hyperreactivity drug therapy, Bronchial Hyperreactivity metabolism, Bronchoalveolar Lavage Fluid chemistry, Cytokines metabolism, Disease Models, Animal, Female, Inflammation drug therapy, Lung drug effects, Male, Methacholine Chloride pharmacology, Mice, Mice, Knockout, Mice, Transgenic, Pulmonary Eosinophilia drug therapy, Pulmonary Eosinophilia metabolism, RNA Interference drug effects, Respiratory Hypersensitivity drug therapy, Allergens metabolism, Inflammation metabolism, Lung metabolism, Membrane Proteins metabolism, RNA, Small Interfering metabolism, Respiratory Hypersensitivity metabolism
- Abstract
Aim/Purpose: Exposure to various allergens has been shown to increase expression of ORMDL3 in the lung in models of allergic asthma. Studies using genetically modified (transgenic or knock out) mice have revealed some of the functions of ORMDL3 in asthma pathogenesis, although amid debate. The goal of this study was to use targeted post-transcriptional downregulation of ORMDL3 in allergen-challenged wild-type (WT) mice by RNA interference to further elucidate the functional role of ORMDL3 in asthma pathogenesis and evaluate a potential therapeutic option. Methods: Allergen (ovalbumin [OVA])-challenged WT mice were administered intranasally (i.n) with a single dose of five short hairpin RNA (shRNA) constructs with different target sequence for murine ORMDL3 cloned in a lentiviral vector or with the empty vector (control). Mice were evaluated for allergen-induced airway hyperresponsiveness (AHR) and various features of airway inflammation after 72 hours. Results: I.n administration of a single dose of ORMDL3 shRNAs to OVA-challenged mice resulted in reduction of ORMDL3 gene expression in the lungs associated with a significant reduction in AHR to inhaled methacholine and in the number of inflammatory cells recruited in the airways, specifically eosinophils, as well as in airway mucus secretion compared to OVA-challenged mice that received the empty vector. Administration of ORMDL3 shRNAs also significantly inhibited levels of IL-13, eotaxin-2 and sphingosine in the lungs. Additionally, ORMDL3 shRNAs significantly inhibited the allergen-mediated increase in monohexyl ceramides C22:0 and C24:0. Conclusions: Post-transcriptional down regulation of ORMDL3 in allergic lungs using i.n-delivered ORMDL3 shRNA (akin to inhaled therapy) attenuates development of key features of airway allergic disease, confirming the involvement of ORMDL3 in allergic asthma pathogenesis and serving as a model for a potential therapeutic strategy.
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- 2020
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11. Detecting direct oral anticoagulants in trauma patients using liquid chromatography-mass spectrometry: A novel approach to medication reconciliation.
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Jayaraman S, DeAntonio JH, Leichtle SW, Han J, Liebrecht L, Contaifer D, Young C, Chou C, Staschen J, Doan D, Kumar NG, Wolfe L, Nguyen T, Chenault G, Anand RJ, Bennett JD, Ferrada P, Goldberg S, Procter LD, Rodas EB, Rossi AP, Whelan JF, Feeser VR, Vitto MJ, Broering B, Hobgood S, Mangino M, Aboutanos M, Bachmann L, and Wijesinghe DS
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- Administration, Oral, Aged, Anticoagulants administration & dosage, Chromatography, High Pressure Liquid, Dabigatran administration & dosage, Dabigatran blood, Female, Healthy Volunteers, Humans, Male, Prospective Studies, Pyrazoles administration & dosage, Pyrazoles blood, Pyridones administration & dosage, Pyridones blood, Rivaroxaban administration & dosage, Rivaroxaban blood, Sensitivity and Specificity, Anticoagulants blood, Mass Spectrometry, Medication Reconciliation methods, Wounds and Injuries blood
- Abstract
Background: Accurate medication reconciliation in trauma patients is essential but difficult. Currently, there is no established clinical method of detecting direct oral anticoagulants (DOACs) in trauma patients. We hypothesized that a liquid chromatography-mass spectrometry (LCMS)-based assay can be used to accurately detect DOACs in trauma patients upon hospital arrival., Methods: Plasma samples were collected from 356 patients who provided informed consent including 10 healthy controls, 19 known positive or negative controls, and 327 trauma patients older than 65 years who were evaluated at our large, urban level 1 trauma center. The assay methodology was developed in healthy and known controls to detect apixaban, rivaroxaban, and dabigatran using LCMS and then applied to 327 samples from trauma patients. Standard medication reconciliation processes in the electronic medical record documenting DOAC usage were compared with LCMS results to determine overall accuracy, sensitivity, specificity, and positive and negative predictive values (PPV, NPV) of the assay., Results: Of 356 patients, 39 (10.96%) were on DOACs: 21 were on apixaban, 14 on rivaroxaban, and 4 on dabigatran. The overall accuracy of the assay for detecting any DOAC was 98.60%, with a sensitivity of 94.87% and specificity of 99.05% (PPV, 92.50%; NPV, 99.37%). The assay detected apixaban with a sensitivity of 90.48% and specificity of 99.10% (PPV, 86.36%; NPV 99.40%). There were three false-positive results and two false-negative LCMS results for apixaban. Dabigatran and rivaroxaban were detected with 100% sensitivity and specificity., Conclusion: This LCMS-based assay was highly accurate in detecting DOACs in trauma patients. Further studies need to confirm the clinical efficacy of this LCMS assay and its value for medication reconciliation in trauma patients., Level of Evidence: Diagnostic Test, level III.
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- 2020
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12. Medication Reconciliation and Patient Safety in Trauma: Applicability of Existing Strategies.
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DeAntonio JH, Leichtle SW, Hobgood S, Boomer L, Aboutanos M, Mangino MJ, Wijesinghe DS, and Jayaraman S
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- Age Factors, Aged, Child, Electronic Health Records organization & administration, Humans, Patient-Centered Care organization & administration, Pharmacists organization & administration, Pharmacy Technicians organization & administration, Professional Role, United States, Medication Reconciliation organization & administration, Patient Safety, Trauma Centers organization & administration, Wounds and Injuries therapy
- Abstract
The Joint Commission has established medication reconciliation as a National Patient Safety Goal, but it has not been studied much in trauma even though it is integral to safe patient care. This article reviews the existing medication reconciliation strategies and their applicability to the trauma setting. To perform medication reconciliation, hospitals use a variety of strategies including pharmacists or pharmacy technicians, electronic medical record tools, and patient-centered strategies. All of these strategies are limited in trauma. Subpopulations such as injured children, the elderly, and those with brain trauma are particularly challenging and are at risk for suboptimal care from inaccurate medication reconciliation. Further research is necessary to create a safe and efficient system for trauma patients., (Copyright © 2019 Elsevier Inc. All rights reserved.)
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- 2020
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13. Plasma ceramide is increased and associated with proteinuria in women with pre-eclampsia and HELLP syndrome.
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Amraoui F, Hassani Lahsinoui H, Spijkers LJA, Vogt L, Peters SLM, Wijesinghe DS, Warncke UO, Chalfant CE, Ris-Stalpers C, van den Born BH, and Afink GB
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- Adult, Case-Control Studies, Female, Gestational Age, Humans, Placenta metabolism, Platelet Count, Pregnancy, Ceramides metabolism, HELLP Syndrome blood, Pre-Eclampsia blood, Proteinuria blood
- Abstract
Objectives: Ceramide is a sphingolipid with anti-angiogenic and pro-apoptotic properties that has shown to be increased in plasma of women with pre-eclampsia. We aimed to compare plasma and placental sphingolipid content among normotensive pregnant women and pre-eclamptic women with and without HELLP syndrome and we aimed to assess whether ceramide is related to hypertension and proteinuria in pre-eclampsia., Study Design: Case-control study. Participants were recruited from the Department of Obstetrics at the Academic Medical Center in Amsterdam, The Netherlands. In total 48 pregnant women were included: 24 with pre-eclampsia and 24 normotensive controls. Of the 24 pre-eclamptic women, 11 had HELLP syndrome., Main Outcome Measures: Plasma and placental ceramide content and correlation with blood pressure and protein excretion in pre-eclampsia., Results: Total plasma, but not placental, ceramide was higher in pre-eclamptic women with HELLP syndrome (11200 95% CI 9531-12870 nmol/ml, n = 11) compared to pre-eclamptic women without HELLP (7413 95% CI 5928-8898 nmol/ml, n = 13, p < 0.001) and normotensive pregnant women (7404 95% CI 6695-8112 nmol/ml, n = 24, p < 0.001). Maternal circulating ceramide levels were strongly associated with proteinuria (r = 0.621, n = 24, p = 0.001) in pre-eclamptic women and inversely correlated with gestational age at delivery (r = 0.771, p < 0.01) in pre-eclamptic women with HELLP syndrome. Plasma ceramide was not correlated with blood pressure., Conclusion: Plasma but not placental ceramide content is increased in women with pre-eclampsia and HELLP syndrome. The strong positive correlation with proteinuria and the inverse correlation with gestational age at delivery indicate that excess plasma ceramide may contribute to the pathophysiology of pre-eclampsia and HELLP., (Copyright © 2019 International Society for the Study of Hypertension in Pregnancy. Published by Elsevier B.V. All rights reserved.)
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- 2020
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14. Dietary Bioactive Fatty Acids as Modulators of Immune Function: Implications on Human Health.
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Kumar NG, Contaifer D, Madurantakam P, Carbone S, Price ET, Van Tassell B, Brophy DF, and Wijesinghe DS
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- Cytokines biosynthesis, Dietary Supplements, Docosahexaenoic Acids administration & dosage, Eicosapentaenoic Acid administration & dosage, Fish Oils administration & dosage, Health Promotion, Humans, Immunity physiology, Lymphocyte Activation drug effects, Receptors, Cell Surface physiology, Dietary Fats, Unsaturated administration & dosage, Fatty Acids, Unsaturated administration & dosage, Immunity drug effects, Inflammation prevention & control
- Abstract
Diet is major modifiable risk factor for cardiovascular disease that can influence the immune status of the individual and contribute to persistent low-grade inflammation. In recent years, there has been an increased appreciation of the role of polyunsaturated fatty acids (PUFA) in improving immune function and reduction of systemic inflammation via the modulation of pattern recognition receptors (PRR) on immune cells. Extensive research on the use of bioactive lipids such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) and their metabolites have illustrated the importance of these pro-resolving lipid mediators in modulating signaling through PRRs. While their mechanism of action, bioavailability in the blood, and their efficacy for clinical use forms an active area of research, they are found widely administered as marine animal-based supplements like fish oil and krill oil to promote health. The focus of this review will be to discuss the effect of these bioactive fatty acids and their metabolites on immune cells and the resulting inflammatory response, with a brief discussion about modern methods for their analysis using mass spectrometry-based methods.
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- 2019
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15. Effect Of Dual sEH/COX-2 Inhibition on Allergen-Induced Airway Inflammation.
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Dileepan M, Rastle-Simpson S, Greenberg Y, Wijesinghe DS, Kumar NG, Yang J, Hwang SH, Hammock BD, Sriramarao P, and Rao SP
- Abstract
Arachidonic acid metabolites resulting from the cyclooxygenase (COX), lipoxygenase, and cytochrome P450 oxidase enzymatic pathways play pro- and anti-inflammatory roles in allergic airway inflammation (AAI) and asthma. Expression of COX-2 and soluble epoxide hydrolase (sEH) are elevated in allergic airways and their enzymatic products (e.g., prostaglandins and diols of epoxyeicosatrienoic acids, respectively) have been shown to participate in the pathogenesis of AAI. Here, we evaluated the outcome of inhibiting the COX-2 and sEH enzymatic pathways with a novel dual inhibitor, PTUPB, in A. alternata -induced AAI. Allergen-challenged mice were administered with 10 or 30 mg/kg of PTUPB, celecoxib (selective COX-2 inhibitor), t -TUCB (selective sEH inhibitor) or vehicle daily by gavage and evaluated for various features of AAI. PTUPB and t -TUCB at 30 mg/kg, but not celecoxib, inhibited eosinophilic infiltration and significantly increased levels of anti-inflammatory EETs in the lung tissue of allergen-challenged mice. t -TUCB significantly inhibited allergen-induced IL-4 and IL-13, while a less pronounced reduction was noted with PTUPB and celecoxib. Additionally, t -TUCB markedly inhibited eotaxin-2, an eosinophil-specific chemokine, which was only marginally reduced by PTUPB and remained elevated in celecoxib-treated mice. PTUPB or t -TUCB administration reversed allergen-induced reduction in levels of various lipid mediators in the lungs, with only a minimal effect noted with celecoxib. Despite the anti-inflammatory effects, PTUPB or t -TUCB did not reduce allergen-induced airway hyperresponsiveness (AHR). However, development of structural changes in the allergic airways, such as mucus hypersecretion and smooth muscle hypertrophy, was significantly inhibited by both inhibitors. Celecoxib, on the other hand, inhibited only airway smooth muscle hypertrophy, but not mucus hypersecretion. In conclusion, dual inhibition of COX-2 and sEH offers no additional advantage relative to sEH inhibition alone in attenuating various features associated with A. alternata -induced AAI, while COX-2 inhibition exerts only moderate or no effect on several of these features. Dual sEH/COX-2 inhibition may be useful in treating conditions where eosinophilic inflammation co-exists with pain-associated inflammation., (Copyright © 2019 Dileepan, Rastle-Simpson, Greenberg, Wijesinghe, Kumar, Yang, Hwang, Hammock, Sriramarao and Rao.)
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- 2019
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16. A Preliminary Investigation towards the Risk Stratification of Allogeneic Stem Cell Recipients with Respect to the Potential for Development of GVHD via Their Pre-Transplant Plasma Lipid and Metabolic Signature.
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Contaifer D Jr, Roberts CH, Kumar NG, Natarajan R, Fisher BJ, Leslie K, Reed J, Toor AA, and Wijesinghe DS
- Abstract
The clinical outcome of allogeneic hematopoietic stem cell transplantation (SCT) may be influenced by the metabolic status of the recipient following conditioning, which in turn may enable risk stratification with respect to the development of transplant-associated complications such as graft vs. host disease (GVHD). To better understand the impact of the metabolic profile of transplant recipients on post-transplant alloreactivity, we investigated the metabolic signature of 14 patients undergoing myeloablative conditioning followed by either human leukocyte antigen (HLA)-matched related or unrelated donor SCT, or autologous SCT. Blood samples were taken following conditioning and prior to transplant on day 0 and the plasma was comprehensively characterized with respect to its lipidome and metabolome via liquid chromatography/mass spectrometry (LCMS) and gas chromatography/mass spectrometry (GCMS). A pro-inflammatory metabolic profile was observed in patients who eventually developed GVHD. Five potential pre-transplant biomarkers, 2-aminobutyric acid, 1-monopalmitin, diacylglycerols (DG 38:5, DG 38:6), and fatty acid FA 20:1 demonstrated high sensitivity and specificity towards predicting post-transplant GVHD. The resulting predictive model demonstrated an estimated predictive accuracy of risk stratification of 100%, with area under the curve of the ROC of 0.995. The likelihood ratio of 1-monopalmitin (infinity), DG 38:5 (6.0), and DG 38:6 (6.0) also demonstrated that a patient with a positive test result for these biomarkers following conditioning and prior to transplant will be at risk of developing GVHD. Collectively, the data suggest the possibility that pre-transplant metabolic signature may be used for risk stratification of SCT recipients with respect to development of alloreactivity.
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- 2019
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17. Metabolic modulation predicts heart failure tests performance.
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Contaifer D Jr, Buckley LF, Wohlford G, Kumar NG, Morriss JM, Ranasinghe AD, Carbone S, Canada JM, Trankle C, Abbate A, Van Tassell BW, and Wijesinghe DS
- Subjects
- Biomarkers blood, Cohort Studies, Female, Humans, Male, Metabolome, Middle Aged, Natriuretic Peptide, Brain blood, Oxygen Consumption, Regression Analysis, Exercise Test, Heart Failure blood, Heart Failure physiopathology
- Abstract
The metabolic changes that accompany changes in Cardiopulmonary testing (CPET) and heart failure biomarkers (HFbio) are not well known. We undertook metabolomic and lipidomic phenotyping of a cohort of heart failure (HF) patients and utilized Multiple Regression Analysis (MRA) to identify associations to CPET and HFBio test performance (peak oxygen consumption (Peak VO2), oxygen uptake efficiency slope (OUES), exercise duration, and minute ventilation-carbon dioxide production slope (VE/VCO2 slope), as well as the established HF biomarkers of inflammation C-reactive protein (CRP), beta-galactoside-binding protein (galectin-3), and N-terminal prohormone of brain natriuretic peptide (NT-proBNP)). A cohort of 49 patients with a left ventricular ejection fraction < 50%, predominantly males African American, presenting a high frequency of diabetes, hyperlipidemia, and hypertension were used in the study. MRA revealed that metabolic models for VE/VCO2 and Peak VO2 were the most fitted models, and the highest predictors' coefficients were from Acylcarnitine C18:2, palmitic acid, citric acid, asparagine, and 3-hydroxybutiric acid. Metabolic Pathway Analysis (MetPA) used predictors to identify the most relevant metabolic pathways associated to the study, aminoacyl-tRNA and amino acid biosynthesis, amino acid metabolism, nitrogen metabolism, pantothenate and CoA biosynthesis, sphingolipid and glycerolipid metabolism, fatty acid biosynthesis, glutathione metabolism, and pentose phosphate pathway (PPP). Metabolite Set Enrichment Analysis (MSEA) found associations of our findings with pre-existing biological knowledge from studies of human plasma metabolism as brain dysfunction and enzyme deficiencies associated with lactic acidosis. Our results indicate a profile of oxidative stress, lactic acidosis, and metabolic syndrome coupled with mitochondria dysfunction in patients with HF tests poor performance. The insights resulting from this study coincides with what has previously been discussed in existing literature thereby supporting the validity of our findings while at the same time characterizing the metabolic underpinning of CPET and HFBio., Competing Interests: The authors have declared that no competing interests exist.
- Published
- 2019
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18. A Literature Based Discovery Visualization System with Hierarchical Clustering and Linking Set Associations.
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Henry S, Panahi A, Wijesinghe DS, and McInnes BT
- Abstract
Literature Based discovery (LBD) seeks to find information implicit in text, but never explicitly stated. In this work, we develop a method of visually summarizing LBD output in an automatically generated tree structure. This structure promotes a comprehensive understanding of LBD output as a whole, and encourages the user to explore branches of the hierarchy they find most interesting or surprising. This novel visualization system requires the development and integration of automatic functional group discovery, set associations, and linking set associations. Specifically, we perform hierarchical clustering on the potential discoveries generated by an LBD system to create a tree of potential hypotheses. We weight the tree by developing set association measures, and extending them to linking set association measures. This weighted tree is displayed in an interactive visual environment, and validated by replicating the historic Raynaud's Disease - fish oil discovery.
- Published
- 2019
19. Implementation of an Active-Learning Laboratory on Pharmacogenetics.
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Powers KE, Buffington TM, Contaifer D Jr, Wijesinghe DS, and Donohoe KL
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- Clinical Competence, Curriculum, Educational Measurement, Humans, Knowledge, Patient Simulation, Program Evaluation, Students, Pharmacy, Education, Pharmacy methods, Education, Pharmacy trends, Pharmacogenetics education, Problem-Based Learning methods
- Abstract
Objective. To evaluate students' knowledge, confidence, and skills after implementation of an active-learning laboratory session in clinical pharmacogenetics. Methods. Third-year pharmacy students (n=130) participated in an active-learning laboratory session on pharmacogenetics. In the laboratory activity, students evaluated patients' pharmacogenetic profiles and documented recommendations to providers based on their findings. Students also counseled a simulated patient on the interpretation of their pharmacogenetic profile. Students' knowledge and confidence were assessed before a lecture on clinical pharmacogenetics, after the lecture, and then after the laboratory activity. The assessment included 10 knowledge-based questions and five confidence questions regarding clinical pharmacogenetics. An evaluation of the laboratory activity was completed after the session. Results. On average, students correctly answered 70.3% of the knowledge-based questions before the lecture, 82.8% after the lecture, and 88.7% after the laboratory session. Additionally, students' confidence improved in each of the five areas assessed. Based on evaluations (response rate: 98.5%), students found that the laboratory activity contributed to their professional development, was taught at an appropriate level for their understanding, and was relevant to pharmacy practice. Conclusion. An active-learning laboratory session to teach pharmacy students about clinical pharmacogenetics improved students' knowledge, confidence, and skills.
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- 2019
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20. Blockade of MCU-Mediated Ca 2+ Uptake Perturbs Lipid Metabolism via PP4-Dependent AMPK Dephosphorylation.
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Tomar D, Jaña F, Dong Z, Quinn WJ 3rd, Jadiya P, Breves SL, Daw CC, Srikantan S, Shanmughapriya S, Nemani N, Carvalho E, Tripathi A, Worth AM, Zhang X, Razmpour R, Seelam A, Rhode S, Mehta AV, Murray M, Slade D, Ramirez SH, Mishra P, Gerhard GS, Caplan J, Norton L, Sharma K, Rajan S, Balciunas D, Wijesinghe DS, Ahima RS, Baur JA, and Madesh M
- Subjects
- AMP-Activated Protein Kinase Kinases, Animals, Calcium Channels genetics, Cells, Cultured, Female, Hep G2 Cells, Humans, Male, Mice, Mice, Inbred C57BL, Mitochondria, Liver metabolism, Mitochondrial Proteins genetics, Phosphoprotein Phosphatases metabolism, Protein Kinases metabolism, Zebrafish, Calcium metabolism, Calcium Channels metabolism, Hepatocytes metabolism, Lipid Metabolism, Mitochondrial Proteins metabolism
- Abstract
Mitochondrial Ca
2+ uniporter (MCU)-mediated Ca2+ uptake promotes the buildup of reducing equivalents that fuel oxidative phosphorylation for cellular metabolism. Although MCU modulates mitochondrial bioenergetics, its function in energy homeostasis in vivo remains elusive. Here we demonstrate that deletion of the Mcu gene in mouse liver (MCUΔhep ) and in Danio rerio by CRISPR/Cas9 inhibits mitochondrial Ca2+ (m Ca2+ ) uptake, delays cytosolic Ca2+ (c Ca2+ ) clearance, reduces oxidative phosphorylation, and leads to increased lipid accumulation. Elevated hepatic lipids in MCUΔhep were a direct result of extramitochondrial Ca2+ -dependent protein phosphatase-4 (PP4) activity, which dephosphorylates AMPK. Loss of AMPK recapitulates hepatic lipid accumulation without changes in MCU-mediated Ca2+ uptake. Furthermore, reconstitution of active AMPK, or PP4 knockdown, enhances lipid clearance in MCUΔhep hepatocytes. Conversely, gain-of-function MCU promotes rapidm Ca2+ uptake, decreases PP4 levels, and reduces hepatic lipid accumulation. Thus, our work uncovers an MCU/PP4/AMPK molecular cascade that links Ca2+ dynamics to hepatic lipid metabolism., (Copyright © 2019 The Authors. Published by Elsevier Inc. All rights reserved.)- Published
- 2019
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21. Medications and patient safety in the trauma setting: a systematic review.
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DeAntonio JH, Nguyen T, Chenault G, Aboutanos MB, Anand RJ, Ferrada P, Goldberg S, Leichtle SW, Procter LD, Rodas EB, Rossi AP, Whelan JF, Feeser VR, Vitto MJ, Broering B, Hobgood S, Mangino M, Wijesinghe DS, and Jayaraman S
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- Humans, Medication Errors mortality, Medication Errors prevention & control, Medication Reconciliation methods, Medication Reconciliation standards, Medication Systems trends, Trauma Centers organization & administration, Trauma Centers standards, Medication Systems standards, Patient Safety standards
- Abstract
Background: Medication errors account for the most common adverse events and a significant cause of mortality in the USA. The Joint Commission has required medication reconciliation since 2006. We aimed to survey the literature and determine the challenges and effectiveness of medication reconciliation in the trauma patient population., Materials and Methods: We conducted a systematic review of the literature to determine the effectiveness of medication reconciliation in trauma patients. English language articles were retrieved from PubMed/Medline, CINAHL, and Cochrane Review databases with search terms "trauma OR injury, AND medication reconciliation OR med rec OR med rek, AND effectiveness OR errors OR intervention OR improvements.", Results: The search resulted in 82 articles. After screening for relevance and duplicates, the 43 remaining were further reviewed, and only four articles, which presented results on medication reconciliation in 3041 trauma patients, were included. Two were retrospective and two were prospective. Two showed only 4% accuracy at time of admission with 48% of medication reconciliations having at least one medication discrepancy. There were major differences across the studies prohibiting comparative statistical analysis., Conclusions: Trauma medication reconciliation is important because of the potential for adverse outcomes given the emergent nature of the illness. The few articles published at this time on medication reconciliation in trauma suggest poor accuracy. Numerous strategies have been implemented in general medicine to improve its accuracy, but these have not yet been studied in trauma. This topic is an important but unrecognized area of research in this field., Competing Interests: N/A: systematic reviewN/A: systematic reviewThe authors declare that they have no competing interests.Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
- Published
- 2019
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22. Untargeted lipidomic analysis to broadly characterize the effects of pathogenic and non-pathogenic staphylococci on mammalian lipids.
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Gajenthra Kumar N, Contaifer D Jr, Baker PRS, Ekroos K, Jefferson KK, and Wijesinghe DS
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- Animals, Cattle, Lipid Metabolism physiology, Lipids, Myocardium metabolism, Staphylococcus metabolism
- Abstract
Modification of the host lipidome via secreted enzymes is an integral, but often overlooked aspect of bacterial pathogenesis. In the current era of prevalent antibiotic resistance, knowledge regarding critical host pathogen lipid interactions has the potential for use in developing novel antibacterial agents. While most studies to date on this matter have focused on specific lipids, or select lipid classes, this provides an incomplete picture. Modern methods of untargeted lipidomics have the capacity to overcome these gaps in knowledge and provide a comprehensive understanding of the role of lipid metabolism in the pathogenesis of infections. In an attempt to determine the role of lipid modifying enzymes produced by staphylococci, we exposed bovine heart lipids, a standardized model for the mammalian lipidome, to spent medium from staphylococcal cultures, and analyzed lipid molecular changes by MS/MSALL shotgun lipidomics. We elucidate distinct effects of different staphylococcal isolates, including 4 clinical isolates of the pathogenic species Staphylococcus aureus, a clinical isolate of the normally commensal species S. epidermidis, and the non-pathogenic species S. carnosus. Two highly virulent strains of S. aureus had a more profound effect on mammalian lipids and modified more lipid classes than the other staphylococcal strains. Our studies demonstrate the utility of the applied untargeted lipidomics methodology to profile lipid changes induced by different bacterial secretomes. Finally, we demonstrate the promise of this lipidomics approach in assessing the specificity of bacterial enzymes for mammalian lipid classes. Our data suggests that there may be a correlation between the bacterial expression of lipid-modifying enzymes and virulence, and could facilitate the guided discovery of lipid pathways required for bacterial infections caused by S. aureus and thereby provide insights into the generation of novel antibacterial agents., Competing Interests: PRSB was employed by SCIEX LLC. during the course of the study. SCIEX allowed PRSB to generate early proof of concept data using their in house instrument platform. PRSB is currently an employee at Avanti Polar Lipids. Lipidomics Consulting Ltd is solely owned and operated by KE. There are no patents, products in development or marketed products to declare. The commercial affiliations of the authors PRSB and KE does not alter our adherence to all the PLOS ONE policies on sharing data and materials. None of the other authors have any competing interests. All the data has been made available with this manuscript.
- Published
- 2018
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23. Erratum. Intensive Versus Standard Blood Pressure Control in SPRINT-Eligible Participants of ACCORD-BP. Diabetes Care 2017;40:1733-1738.
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Buckley LF, Dixon DL, Wohlford GF 4th, Wijesinghe DS, Baker WL, and Van Tassell BW
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- 2018
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24. Metabolomics and Precision Medicine in Trauma: The State of the Field.
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Jayaraman SP, Anand RJ, DeAntonio JH, Mangino M, Aboutanos MB, Kasirajan V, Ivatury RR, Valadka AB, Glushakova O, Hayes RL, Bachmann LM, Brophy GM, Contaifer D, Warncke UO, Brophy DF, and Wijesinghe DS
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- Humans, Wounds and Injuries blood, Wounds and Injuries metabolism, Metabolomics methods, Precision Medicine methods
- Abstract
Trauma is a major problem in the United States. Mortality from trauma is the number one cause of death under the age of 45 in the United States and is the third leading cause of death for all age groups. There are approximately 200,000 deaths per year due to trauma in the United States at a cost of over $671 billion in combined healthcare costs and lost productivity. Unsurprisingly, trauma accounts for approximately 30% of all life-years lost in the United States. Due to immense development of trauma systems, a large majority of trauma patients survive the injury, but then go on to die from complications arising from the injury. These complications are marked by early and significant metabolic changes accompanied by inflammatory responses that lead to progressive organ failure and, ultimately, death. Early resuscitative and surgical interventions followed by close monitoring to identify and rescue treatment failures are key to successful outcomes. Currently, the adequacy of resuscitation is measured using vital signs, noninvasive methods such as bedside echocardiography or stroke volume variation, and other laboratory endpoints of resuscitation, such as lactate and base deficit. However, these methods may be too crude to understand cellular and subcellular changes that may be occurring in trauma patients. Better diagnostic and therapeutic markers are needed to assess the adequacy of interventions and monitor responses at a cellular and subcellular level and inform clinical decision-making before complications are clinically apparent. The developing field of metabolomics holds great promise in the identification and application of biochemical markers toward the clinical decision-making process.
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- 2018
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25. Response to Comment on Buckley et al. Intensive Versus Standard Blood Pressure Control in SPRINT-Eligible Participants of ACCORD-BP. Diabetes Care 2017;40:1733-1738.
- Author
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Buckley LF, Dixon DL, Wohlford GF 4th, Wijesinghe DS, Baker WL, and Van Tassell BW
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- Blood Pressure drug effects, Humans, Antihypertensive Agents, Hypertension
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- 2018
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26. Effect of intensive blood pressure control in patients with type 2 diabetes mellitus over 9 years of follow-up: A subgroup analysis of high-risk ACCORDION trial participants.
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Buckley LF, Dixon DL, Wohlford GF 4th, Wijesinghe DS, Baker WL, and Van Tassell BW
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- Aged, Angina, Unstable etiology, Angina, Unstable mortality, Diabetes Mellitus, Type 2 complications, Diabetes Mellitus, Type 2 mortality, Diabetic Angiopathies complications, Diabetic Angiopathies mortality, Double-Blind Method, Female, Follow-Up Studies, Heart Failure etiology, Heart Failure mortality, Humans, Hypertension complications, Hypertension mortality, Male, Middle Aged, Myocardial Infarction etiology, Myocardial Infarction mortality, Stroke etiology, Stroke mortality, Antihypertensive Agents therapeutic use, Diabetes Mellitus, Type 2 prevention & control, Diabetic Angiopathies prevention & control, Hypertension prevention & control
- Abstract
Although guidelines recommend strict blood pressure (BP) control in patients with type 2 diabetes mellitus (T2DM) and elevated cardiovascular risk, the long-term effects of this approach are unknown. We investigated the effect of intensive BP control on clinical outcomes in patients with T2DM over 9 years of follow-up. We included Action to Control Cardiovascular Risk in Diabetes - Blood Pressure participants in the standard glucose control arm who had established cardiovascular disease, chronic kidney disease, were ≥75 years of age or who had a 10-year coronary heart risk ≥15%. Participants were randomized to either intensive (systolic BP < 120 mm Hg) or standard (systolic BP < 140 mm Hg) BP control for an average of 5 years. Observational follow-up occurred for an average of 4 years thereafter. After an average total follow-up of 9 years, intensive BP control reduced the composite of cardiovascular death, nonfatal myocardial infarction and nonfatal stroke by 25% (hazard ratio, 0.75; 95% confidence interval, 0.60-0.95; P = .02). The overall benefit was driven by a reduction in nonfatal myocardial infarction (P = .01). In this post-hoc analysis, the benefits of a fixed-duration intensive BP control intervention in patients with T2DM persisted throughout 9 years of follow-up., (© 2018 John Wiley & Sons Ltd.)
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- 2018
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27. Intensive Versus Standard Blood Pressure Control in SPRINT-Eligible Participants of ACCORD-BP.
- Author
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Buckley LF, Dixon DL, Wohlford GF 4th, Wijesinghe DS, Baker WL, and Van Tassell BW
- Subjects
- Aged, Blood Pressure drug effects, Diabetes Mellitus, Type 2 complications, Diabetes Mellitus, Type 2 mortality, Female, Heart Failure etiology, Heart Failure mortality, Heart Failure prevention & control, Humans, Hypertension mortality, Male, Middle Aged, Myocardial Infarction etiology, Myocardial Infarction mortality, Myocardial Infarction prevention & control, Proportional Hazards Models, Risk Factors, Stroke etiology, Stroke mortality, Stroke prevention & control, Treatment Outcome, Antihypertensive Agents therapeutic use, Hypertension drug therapy
- Abstract
Objective: We sought to determine the effect of intensive blood pressure (BP) control on cardiovascular outcomes in participants with type 2 diabetes mellitus (T2DM) and additional risk factors for cardiovascular disease (CVD)., Research Design and Methods: This study was a post hoc, multivariate, subgroup analysis of ACCORD-BP (Action to Control Cardiovascular Risk in Diabetes Blood Pressure) participants. Participants were eligible for the analysis if they were in the standard glucose control arm of ACCORD-BP and also had the additional CVD risk factors required for SPRINT (Systolic Blood Pressure Intervention Trial) eligibility. We used a Cox proportional hazards regression model to compare the effect of intensive versus standard BP control on CVD outcomes. The "SPRINT-eligible" ACCORD-BP participants were pooled with SPRINT participants to determine whether the effects of intensive BP control interacted with T2DM., Results: The mean baseline Framingham 10-year CVD risk scores were 14.5% and 14.8%, respectively, in the intensive and standard BP control groups. The mean achieved systolic BP values were 120 and 134 mmHg in the intensive and standard BP control groups ( P < 0.001). Intensive BP control reduced the composite of CVD death, nonfatal myocardial infarction (MI), nonfatal stroke, any revascularization, and heart failure (hazard ratio 0.79; 95% CI 0.65-0.96; P = 0.02). Intensive BP control also reduced CVD death, nonfatal MI, and nonfatal stroke (hazard ratio 0.69; 95% CI 0.51-0.93; P = 0.01). Treatment-related adverse events occurred more frequently in participants receiving intensive BP control (4.1% vs. 2.1%; P = 0.003). The effect of intensive BP control on CVD outcomes did not differ between patients with and without T2DM ( P > 0.62)., Conclusions: Intensive BP control reduced CVD outcomes in a cohort of participants with T2DM and additional CVD risk factors., (© 2017 by the American Diabetes Association.)
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- 2017
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28. Circulating Lipids and Acute Pain Sensitization: An Exploratory Analysis.
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Starkweather A, Julian T, Ramesh D, Heineman A, Sturgill J, Dorsey SG, Lyon DE, and Wijesinghe DS
- Subjects
- Acute Disease, Adult, Female, Humans, Low Back Pain prevention & control, Male, Mass Spectrometry, Middle Aged, Musculoskeletal Pain blood, Pain Measurement methods, Risk Factors, Central Nervous System Sensitization physiology, Low Back Pain blood, Triglycerides blood
- Abstract
Background: In individuals with low back pain, higher lipid levels have been documented and were associated with increased risk for chronic low back pain., Objectives: The purpose of this research was to identify plasma lipids that discriminate participants with acute low back pain with or without pain sensitization as measured by quantitative sensory testing., Methods: This exploratory study was conducted as part of a larger parent randomized controlled trial. A cluster analysis of 30 participants with acute low back pain revealed two clusters: one with signs of peripheral and central sensitivity to mechanical and thermal stimuli and the other with an absence of peripheral and central sensitivity. Lipid levels were extracted from plasma and measured using mass spectroscopy., Results: Triacylglycerol 50:2 was significantly higher in participants with peripheral and central sensitization compared to the nonsensitized cluster. The nonsensitized cluster had significantly higher levels of phosphoglyceride 34:2, plasmenyl phosphocholine 38:1, and phosphatidic acid 28:1 compared to participants with peripheral and central sensitization. Linear discriminant function analysis was conducted using the four statistically significant lipids to test their predictive power to classify those in the sensitization and no-sensitization clusters; the four lipids accurately predicted cluster classification 58% of the time (R = .58, -2 log likelihood = 14.59)., Discussion: The results of this exploratory study suggest a unique lipidomic signature in plasma of patients with acute low back pain based on the presence or absence of pain sensitization. Future work to replicate these preliminary findings is underway.
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- 2017
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29. Prospective clinical biomarkers of caspase-mediated apoptosis associated with neuronal and neurovascular damage following stroke and other severe brain injuries: Implications for chronic neurodegeneration.
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Glushakova OY, Glushakov AA, Wijesinghe DS, Valadka AB, Hayes RL, and Glushakov AV
- Abstract
Acute brain injuries, including ischemic and hemorrhagic stroke, as well as traumatic brain injury (TBI), are major worldwide health concerns with very limited options for effective diagnosis and treatment. Stroke and TBI pose an increased risk for the development of chronic neurodegenerative diseases, notably chronic traumatic encephalopathy, Alzheimer's disease, and Parkinson's disease. The existence of premorbid neurodegenerative diseases can exacerbate the severity and prognosis of acute brain injuries. Apoptosis involving caspase-3 is one of the most common mechanisms involved in the etiopathology of both acute and chronic neurological and neurodegenerative diseases, suggesting a relationship between these disorders. Over the past two decades, several clinical biomarkers of apoptosis have been identified in cerebrospinal fluid and peripheral blood following ischemic stroke, intracerebral and subarachnoid hemorrhage, and TBI. These biomarkers include selected caspases, notably caspase-3 and its specific cleavage products such as caspase-cleaved cytokeratin-18, caspase-cleaved tau, and a caspase-specific 120 kDa αII-spectrin breakdown product. The levels of these biomarkers might be a valuable tool for the identification of pathological pathways such as apoptosis and inflammation involved in injury progression, assessment of injury severity, and prediction of clinical outcomes. This review focuses on clinical studies involving biomarkers of caspase-3-mediated pathways, following stroke and TBI. The review further examines their prospective diagnostic utility, as well as clinical utility for improved personalized treatment of stroke and TBI patients and the development of prophylactic treatment chronic neurodegenerative disease., Competing Interests: Ronald L. Hayes owns stock, receives compensation from and is an executive officer of Banyan Biomarkers, Inc., and, as such, may benefit financially as a result of the outcomes of this research or work reported in this publication.
- Published
- 2017
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30. Unsupervised analysis of combined lipid and coagulation data reveals coagulopathy subtypes among dialysis patients.
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Contaifer D Jr, Carl DE, Warncke UO, Martin EJ, Mohammed BM, Van Tassell B, Brophy DF, Chalfant CE, and Wijesinghe DS
- Subjects
- Adult, Biomarkers blood, Blood Coagulation, Blood Coagulation Disorders etiology, Blood Coagulation Disorders pathology, Chromatography, High Pressure Liquid, Female, Humans, Kidney Failure, Chronic complications, Kidney Failure, Chronic pathology, Kidney Failure, Chronic therapy, Lipid Metabolism, Lysophospholipids metabolism, Male, Middle Aged, Renal Insufficiency, Chronic complications, Renal Insufficiency, Chronic pathology, Renal Insufficiency, Chronic therapy, Sphingolipids metabolism, Sphingomyelins metabolism, Sphingosine analogs & derivatives, Sphingosine metabolism, Blood Coagulation Disorders metabolism, Kidney Failure, Chronic blood, Peritoneal Dialysis adverse effects, Renal Dialysis adverse effects, Renal Insufficiency, Chronic blood
- Abstract
Hemodialysis (HD) and peritoneal dialysis (PD) are the primary means of managing end stage renal disease (ESRD). However, these treatment modalities are associated with the onset of coagulation abnormalities. Effective management of coagulation risk among these patients requires the identification of surrogate markers that provide an early indication of the coagulation abnormalities. The role of sphingolipids in the manifestation and prediction of coagulation abnormalities among dialysis patients have never been investigated. Herein, we report the first instance of an in depth investigation into the sphingolipid changes among ESRD patients undergoing HD and PD. The results reveal distinct differences in terms of perturbations to specific sphingolipid biosynthetic pathways that are highly dependent on the treatment modality. Our studies also demonstrated strong correlation between specific sphingolipids and coagulation parameters, such as HexCer(d18:1/26:0) and maximal amplitude (MA), SM(d18:1/24:1) and tissue factor pathway inhibitor, and sphingosine 1-phosphate d18:1 and FX (Spearman ρ of 0.93, 0.89, and -0.89, respectively). Furthermore, our study revealed the potential for using HexCer(d18:1/22:0), HexCer(d18:1/24:0), and HexCer(d18:1/26:0) (r
2 = 0.71, 0.82, and 0.63, respectively) and coagulation parameter MA (r2 = 0.7) for successful diagnosis of differential coagulopathies among ESRD patients undergoing HD, providing an opportunity toward personalized disease management., (Copyright © 2017 by the American Society for Biochemistry and Molecular Biology, Inc.)- Published
- 2017
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31. Impact of high dose vitamin C on platelet function.
- Author
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Mohammed BM, Sanford KW, Fisher BJ, Martin EJ, Contaifer D Jr, Warncke UO, Wijesinghe DS, Chalfant CE, Brophy DF, Fowler Iii AA, and Natarajan R
- Abstract
Aim: To examine the effect of high doses of vitamin C (VitC) on ex vivo human platelets (PLTs)., Methods: Platelet concentrates collected for therapeutic or prophylactic transfusions were exposed to: (1) normal saline (control); (2) 0.3 mmol/L VitC (Lo VitC); or (3) 3 mmol/L VitC (Hi VitC, final concentrations) and stored appropriately. The VitC additive was preservative-free buffered ascorbic acid in water, pH 5.5 to 7.0, adjusted with sodium bicarbonate and sodium hydroxide. The doses of VitC used here correspond to plasma VitC levels reported in recently completed clinical trials. Prior to supplementation, a baseline sample was collected for analysis. PLTs were sampled again on days 2, 5 and 8 and assayed for changes in PLT function by: Thromboelastography (TEG), for changes in viscoelastic properties; aggregometry, for PLT aggregation and adenosine triphosphate (ATP) secretion in response to collagen or adenosine diphosphate (ADP); and flow cytometry, for changes in expression of CD-31, CD41a, CD62p and CD63. In addition, PLT intracellular VitC content was measured using a fluorimetric assay for ascorbic acid and PLT poor plasma was used for plasma coagulation tests [prothrombin time (PT), partial thrombplastin time (PTT), functional fibrinogen] and Lipidomics analysis (UPLC ESI-MS/MS)., Results: VitC supplementation significantly increased PLTs intracellular ascorbic acid levels from 1.2 mmol/L at baseline to 3.2 mmol/L (Lo VitC) and 15.7 mmol/L (Hi VitC, P < 0.05). VitC supplementation did not significantly change PT and PTT values, or functional fibrinogen levels over the 8 d exposure period ( P > 0.05). PLT function assayed by TEG, aggregometry and flow cytometry was not significantly altered by Lo or Hi VitC for up to 5 d. However, PLTs exposed to 3 mmol/L VitC for 8 d demonstrated significantly increased R and K times by TEG and a decrease in the α-angle ( P < 0.05). There was also a fall of 20 mm in maximum amplitude associated with the Hi VitC compared to both baseline and day 8 saline controls. Platelet aggregation studies, showed uniform declines in collagen and ADP-induced platelet aggregations over the 8-d study period in all three groups ( P > 0.05). Collagen and ADP-induced ATP secretion was also not different between the three groups ( P > 0.05). Finally, VitC at the higher dose (3 mmol/L) also induced the release of several eicosanoids including thromboxane B
2 and prostaglandin E2 , as well as products of arachidonic acid metabolism via the lipoxygenases pathway such as 11-/12-/15-hydroxyicosatetraenoic acid ( P < 0.05)., Conclusion: Alterations in PLT function by exposure to 3 mmol/L VitC for 8 d suggest that caution should be exerted with prolonged use of intravenous high dose VitC., Competing Interests: Conflict-of-interest statement: To the best of the authors’ knowledge, no conflict of interest exists.- Published
- 2017
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32. Human as the Ultimate Wound Healing Model: Strategies for Studies Investigating the Dermal Lipidome.
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Wijesinghe DS, Warncke UO, and Diegelmann RF
- Abstract
Purpose of Review: Educate the reader of the multiple roles undertaken by the human epidermal lipidome and the experimental techniques of measuring them., Recent Findings: Damage to skin elicits a wound healing process that is capped by the recreation of the lipid barrier. In addition to barrier function, lipids also undertake an active signaling role during wound healing. Achievement of these multiple functions necessitates a significant complexity and diversity in the lipidome resulting in a composition that is unique to the human skin. As such, any attempts to delineate the function of the lipidome during the wound healing process in humans need to be addressed via studies undertaken in humans., Summary: The human cutaneous lipidome is unique and play a functionally significant role in maintaining barrier and regulating wound healing. Modern mass spectrometry and Raman spectroscopy based methods enable the investigation epidermal lipidome with respect to those functions., Competing Interests: Conflict of Interest Dayanjan S Wijesinghe, Urszula Osinska Warncke, and Robert F. Diegelmann declare that they have no conflicts of interest.
- Published
- 2016
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33. Multivariate Curve Resolution-Alternating Least Squares Analysis of High-Resolution Liquid Chromatography-Mass Spectrometry Data.
- Author
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Sinanian MM, Cook DW, Rutan SC, and Wijesinghe DS
- Abstract
Methods such as liquid chromatography coupled with high-resolution mass spectrometry (LC-HRMS) are crucial for differentiating compounds with highly similar masses. This is a necessity when analyzing highly complex samples; however, the size of high-resolution LC-HRMS data sets can cause difficulties when applying advanced data analysis techniques. In this work, LC-HRMS analyses of known amphetamine samples and unknown bacterial lipid samples were carried out, and multivariate curve resolution-alternating least squares (MCR-ALS) was applied to the data to obtain mathematical separation of overlapped analyte signals. In order to minimize computational strain, a novel strategy was developed which minimizes the number of irrelevant masses analyzed at full resolution. To do this, data were first binned to unit mass resolution, and MCR-ALS was performed. This provided mathematical components for each analyte present plus background components. In the resolved spectral profiles of analyte components, masses above a preset intensity threshold were extracted, discarding all other masses, and expanded to successively higher levels of resolution, applying MCR-ALS at each level. These steps were repeated until 0.001 amu resolution was achieved, as dictated by the resolution of the instrument-in this case, a time-of-flight mass spectrometer. This strategy allowed for the accurate recovery of all known amphetamine compounds and select bacterial lipid extracts while minimizing the size of the data, therefore minimizing computational analysis time and data storage requirements. This relatively simple strategy enables the effective coupling of LC-HRMS with MCR-ALS.
- Published
- 2016
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34. A diet-induced animal model of non-alcoholic fatty liver disease and hepatocellular cancer.
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Asgharpour A, Cazanave SC, Pacana T, Seneshaw M, Vincent R, Banini BA, Kumar DP, Daita K, Min HK, Mirshahi F, Bedossa P, Sun X, Hoshida Y, Koduru SV, Contaifer D Jr, Warncke UO, Wijesinghe DS, and Sanyal AJ
- Subjects
- Animals, Carcinoma, Hepatocellular, Diet, High-Fat, Disease Models, Animal, Humans, Liver, Liver Neoplasms, Mice, Mice, Inbred C57BL, Non-alcoholic Fatty Liver Disease
- Abstract
Background & Aims: The lack of a preclinical model of progressive non-alcoholic steatohepatitis (NASH) that recapitulates human disease is a barrier to therapeutic development., Methods: A stable isogenic cross between C57BL/6J (B6) and 129S1/SvImJ (S129) mice were fed a high fat diet with ad libitum consumption of glucose and fructose in physiologically relevant concentrations and compared to mice fed a chow diet and also to both parent strains., Results: Following initiation of the obesogenic diet, B6/129 mice developed obesity, insulin resistance, hypertriglyceridemia and increased LDL-cholesterol. They sequentially also developed steatosis (4-8weeks), steatohepatitis (16-24weeks), progressive fibrosis (16weeks onwards) and spontaneous hepatocellular cancer (HCC). There was a strong concordance between the pattern of pathway activation at a transcriptomic level between humans and mice with similar histological phenotypes (FDR 0.02 for early and 0.08 for late time points). Lipogenic, inflammatory and apoptotic signaling pathways activated in human NASH were also activated in these mice. The HCC gene signature resembled the S1 and S2 human subclasses of HCC (FDR 0.01 for both). Only the B6/129 mouse but not the parent strains recapitulated all of these aspects of human NAFLD., Conclusions: We here describe a diet-induced animal model of non-alcoholic fatty liver disease (DIAMOND) that recapitulates the key physiological, metabolic, histologic, transcriptomic and cell-signaling changes seen in humans with progressive NASH., Lay Summary: We have developed a diet-induced mouse model of non-alcoholic steatohepatitis (NASH) and hepatic cancers in a cross between two mouse strains (129S1/SvImJ and C57Bl/6J). This model mimics all the physiological, metabolic, histological, transcriptomic gene signature and clinical endpoints of human NASH and can facilitate preclinical development of therapeutic targets for NASH., Competing Interests: Dr. Sanyal reports grants from NIH, during the conduct of the study; In addition, Dr. Sanyal has a patent VCU pending and Virginia Commonwealth University and Dr. Sanyal has established a biotechnology company (SanyalBio) which may test compounds being evaluated for NASH in the future. All the other authors who have taken part in this study declared that they do not have anything to disclose regarding funding or conflict of interest with respect to this manuscript., (Copyright © 2016 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.)
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- 2016
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35. Fluvastatin Suppresses Mast Cell and Basophil IgE Responses: Genotype-Dependent Effects.
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Kolawole EM, McLeod JJ, Ndaw V, Abebayehu D, Barnstein BO, Faber T, Spence AJ, Taruselli M, Paranjape A, Haque TT, Qayum AA, Kazmi QA, Wijesinghe DS, Sturgill JL, Chalfant CE, Straus DB, Oskeritzian CA, and Ryan JJ
- Subjects
- Acyl Coenzyme A genetics, Acyl Coenzyme A immunology, Animals, Apoptosis, Basophils immunology, Cells, Cultured, Cytokines biosynthesis, Farnesyltranstransferase metabolism, Female, Fluvastatin, Genotype, Humans, Hydroxymethylglutaryl-CoA Reductase Inhibitors pharmacology, Immunoglobulin E immunology, Mast Cells immunology, Mevalonic Acid pharmacology, Mice, Mice, Inbred C57BL, Signal Transduction drug effects, Th2 Cells immunology, Basophils drug effects, Fatty Acids, Monounsaturated pharmacology, Immunoglobulin E biosynthesis, Indoles pharmacology, Mast Cells drug effects
- Abstract
Mast cell (MC)- and basophil-associated inflammatory diseases are a considerable burden to society. A significant portion of patients have symptoms despite standard-of-care therapy. Statins, used to lower serum cholesterol, have immune-modulating activities. We tested the in vitro and in vivo effects of statins on IgE-mediated MC and basophil activation. Fluvastatin showed the most significant inhibitory effects of the six statins tested, suppressing IgE-induced cytokine secretion among mouse MCs and basophils. The effects of fluvastatin were reversed by mevalonic acid or geranylgeranyl pyrophosphatase, and mimicked by geranylgeranyl transferase inhibition. Fluvastatin selectively suppressed key FcεRI signaling pathways, including Akt and ERK. Although MCs and basophils from the C57BL/6J mouse strain were responsive to fluvastatin, those from 129/SvImJ mice were completely resistant. Resistance correlated with fluvastatin-induced upregulation of the statin target HMG-CoA reductase. Human MC cultures from eight donors showed a wide range of fluvastatin responsiveness. These data demonstrate that fluvastatin is a potent suppressor of IgE-mediated MC activation, acting at least partly via blockade of geranyl lipid production downstream of HMG-CoA reductase. Importantly, consideration of statin use for treating MC-associated disease needs to incorporate genetic background effects, which can yield drug resistance., (Copyright © 2016 by The American Association of Immunologists, Inc.)
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- 2016
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36. Anaplasma phagocytophilum Rab10-dependent parasitism of the trans-Golgi network is critical for completion of the infection cycle.
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Truchan HK, VieBrock L, Cockburn CL, Ojogun N, Griffin BP, Wijesinghe DS, Chalfant CE, and Carlyon JA
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- Cell Line, Cytoplasmic Vesicles chemistry, Humans, Nucleoside-Phosphate Kinase metabolism, Protein Binding, Anaplasma phagocytophilum growth & development, Cytoplasmic Vesicles metabolism, Host-Parasite Interactions, Vacuoles microbiology, rab GTP-Binding Proteins analysis, trans-Golgi Network microbiology
- Abstract
Anaplasma phagocytophilum is an emerging human pathogen and obligate intracellular bacterium. It inhabits a host cell-derived vacuole and cycles between replicative reticulate cell (RC) and infectious dense-cored (DC) morphotypes. Host-pathogen interactions that are critical for RC-to-DC conversion are undefined. We previously reported that A. phagocytophilum recruits green fluorescent protein (GFP)-tagged Rab10, a GTPase that directs exocytic traffic from the sphingolipid-rich trans-Golgi network (TGN) to its vacuole in a guanine nucleotide-independent manner. Here, we demonstrate that endogenous Rab10-positive TGN vesicles are not only routed to but also delivered into the A. phagocytophilum-occupied vacuole (ApV). Consistent with this finding, A. phagocytophilum incorporates sphingolipids while intracellular and retains them when naturally released from host cells. TGN vesicle delivery into the ApV is Rab10 dependent, up-regulates expression of the DC-specific marker, APH1235, and is critical for the production of infectious progeny. The A. phagocytophilum surface protein, uridine monophosphate kinase, was identified as a guanine nucleotide-independent, Rab10-specific ligand. These data delineate why Rab10 is important for the A. phagocytophilum infection cycle and expand the understanding of the benefits that exploiting host cell membrane traffic affords intracellular bacterial pathogens., (© 2015 John Wiley & Sons Ltd.)
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- 2016
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37. Arachidonic acid-derived signaling lipids and functions in impaired healing.
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Dhall S, Wijesinghe DS, Karim ZA, Castro A, Vemana HP, Khasawneh FT, Chalfant CE, and Martins-Green M
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- Animals, Disease Models, Animal, Mass Spectrometry, Mice, Mice, Inbred C57BL, Pancreatic Elastase metabolism, Signal Transduction drug effects, Skin metabolism, Skin pathology, Swine, Wounds and Injuries metabolism, Wounds and Injuries pathology, Arachidonic Acid pharmacology, Lipid Metabolism drug effects, Oxidative Stress, Skin injuries, Wound Healing drug effects, Wounds and Injuries drug therapy
- Abstract
Very little is known about lipid function during wound healing, and much less during impaired healing. Such understanding will help identify what roles lipid signaling plays in the development of impaired/chronic wounds. We took a lipidomics approach to study the alterations in lipid profile in the LIGHT(-/-) mouse model of impaired healing which has characteristics that resemble those of impaired/chronic wounds in humans, including high levels of oxidative stress, excess inflammation, increased extracellular matrix degradation and blood vessels with fibrin cuffs. The latter suggests excess coagulation and potentially increased platelet aggregation. We show here that in these impaired wounds there is an imbalance in the arachidonic acid (AA) derived eicosonoids that mediate or modulate inflammatory reactions and platelet aggregation. In the LIGHT(-/-) impaired wounds there is a significant increase in enzymatically derived breakdown products of AA. We found that early after injury there was a significant increase in the eicosanoids 11-, 12-, and 15-hydroxyeicosa-tetranoic acid, and the proinflammatory leukotrienes (LTD4 and LTE) and prostaglandins (PGE2 and PGF2α ). Some of these eicosanoids also promote platelet aggregation. This led us to examine the levels of other eicosanoids known to be involved in the latter process. We found that thromboxane (TXA2 /B2 ), and prostacyclins 6kPGF1α are elevated shortly after wounding and in some cases during healing. To determine whether they have an impact in platelet aggregation and hemostasis, we tested LIGHT(-/-) mouse wounds for these two parameters and found that, indeed, platelet aggregation and hemostasis are enhanced in these mice when compared with the control C57BL/6 mice. Understanding lipid signaling in impaired wounds can potentially lead to development of new therapeutics or in using existing nonsteroidal anti-inflammatory agents to help correct the course of healing., (© 2015 by the Wound Healing Society.)
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- 2015
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38. Increased eicosanoid levels in the Sugen/chronic hypoxia model of severe pulmonary hypertension.
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Al-Husseini A, Wijesinghe DS, Farkas L, Kraskauskas D, Drake JI, Van Tassel B, Abbate A, Chalfant CE, and Voelkel NF
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- Animals, Arachidonate 5-Lipoxygenase genetics, Arachidonate 5-Lipoxygenase metabolism, Arterioles drug effects, Arterioles enzymology, Arterioles pathology, Cyclooxygenase 2 genetics, Cyclooxygenase 2 metabolism, Cyclooxygenase 2 Inhibitors pharmacology, Dinoprost antagonists & inhibitors, Dinoprost biosynthesis, Dinoprostone antagonists & inhibitors, Dinoprostone biosynthesis, Humans, Hypertension, Pulmonary enzymology, Hypertension, Pulmonary genetics, Hypoxia enzymology, Hypoxia genetics, Hypoxia pathology, Inflammation, Leukotriene D4 antagonists & inhibitors, Leukotriene D4 biosynthesis, Lung enzymology, Lung pathology, Male, Prostaglandins F antagonists & inhibitors, Prostaglandins F biosynthesis, Pulmonary Artery drug effects, Pulmonary Artery enzymology, Pulmonary Artery pathology, Pyrazoles pharmacology, Rats, Rats, Sprague-Dawley, Thromboxane B2 antagonists & inhibitors, Thromboxane B2 biosynthesis, Ventricular Function, Right drug effects, Anti-Inflammatory Agents, Non-Steroidal pharmacology, Diethylcarbamazine pharmacology, Hypertension, Pulmonary drug therapy, Hypoxia drug therapy, Lipoxygenase Inhibitors pharmacology, Lung drug effects
- Abstract
Inflammation and altered immunity are recognized components of severe pulmonary arterial hypertension in human patients and in animal models of PAH. While eicosanoid metabolites of cyclooxygenase and lipoxygenase pathways have been identified in the lungs from pulmonary hypertensive animals their role in the pathogenesis of severe angioobliterative PAH has not been examined. Here we investigated whether a cyclooxygenase-2 (COX-2) inhibitor or diethylcarbamazine (DEC), that is known for its 5-lipoxygenase inhibiting and antioxidant actions, modify the development of PAH in the Sugen 5416/hypoxia (SuHx) rat model. The COX-2 inhibitor SC-58125 had little effect on the right ventricular pressure and did not prevent the development of pulmonary angioobliteration. In contrast, DEC blunted the muscularization of pulmonary arterioles and reduced the number of fully obliterated lung vessels. DEC treatment of SuHx rats, after the lung vascular disease had been established, reduced the degree of PAH, the number of obliterated arterioles and the degree of perivascular inflammation. We conclude that the non-specific anti-inflammatory drug DEC affects developing PAH and is partially effective once angioobliterative PAH has been established.
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- 2015
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39. The Lipid Portion of Activated Platelet-Rich Plasma Significantly Contributes to Its Wound Healing Properties.
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Hoeferlin LA, Huynh QK, Mietla JA, Sell SA, Tucker J, Chalfant CE, and Wijesinghe DS
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Objective: Platelet-rich plasma (PRP) is a popular choice for the treatment of chronic wounds. Current dogma attributes these healing properties to the peptide growth factors of PRP. However, PRP is also rich in bioactive lipids whose contribution to healing has not been characterized and warrants investigation due to the protease-rich environment of chronic wounds. Approach: The lipid fraction of PRP was tested with respect to proliferation and migration of primary adult human dermal fibroblasts (HDFa)±exposure to chronic wound fluid (CWF). This fraction was also characterized via LC-MS/MS for bioactive lipids. A synthetic formulation of the bioactive lipid composition was developed and tested for the ability to overcome proliferative growth arrest induced by CWF. Results: The data demonstrate the ability of the lipid fraction of PRP to significantly enhance the migration and proliferation of HDFa, and to overcome the proliferative growth arrest induced by CWF. Furthermore, the synthetic lipid formulation generated following characterization of the PRP lipidome demonstrated a similar ability to overcome proliferative arrest of HDFa in the presence of CWF. Innovation: For the first time, we demonstrate the relevance of the lipid fraction of PRP toward the biology of wound healing. These studies open the possibility of altering the lipid profile of PRP via diet or exogenous pathway manipulation to obtain a better healing outcome. Conclusion: The lipid fraction of PRP is under investigated and yet relevant component in wound healing. The current study demonstrates the relevance of this fraction in wound healing by PRP.
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- 2015
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40. A novel model of chronic wounds: importance of redox imbalance and biofilm-forming bacteria for establishment of chronicity.
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Dhall S, Do D, Garcia M, Wijesinghe DS, Brandon A, Kim J, Sanchez A, Lyubovitsky J, Gallagher S, Nothnagel EA, Chalfant CE, Patel RP, Schiller N, and Martins-Green M
- Subjects
- Animals, Chronic Disease, Disease Models, Animal, Drug Resistance, Bacterial, Mice, Inbred C57BL, Mice, Knockout, Oxidation-Reduction, Oxidative Stress, Reactive Oxygen Species metabolism, Staphylococcal Infections metabolism, Staphylococcal Infections pathology, Staphylococcal Skin Infections metabolism, Staphylococcal Skin Infections pathology, Streptococcal Infections metabolism, Streptococcal Infections pathology, Tumor Necrosis Factor Ligand Superfamily Member 14 genetics, Wound Healing, Biofilms, Staphylococcal Infections microbiology, Staphylococcal Skin Infections microbiology, Streptococcal Infections microbiology
- Abstract
Chronic wounds have a large impact on health, affecting ∼6.5 M people and costing ∼$25B/year in the US alone. We previously discovered that a genetically modified mouse model displays impaired healing similar to problematic wounds in humans and that sometimes the wounds become chronic. Here we show how and why these impaired wounds become chronic, describe a way whereby we can drive impaired wounds to chronicity at will and propose that the same processes are involved in chronic wound development in humans. We hypothesize that exacerbated levels of oxidative stress are critical for initiation of chronicity. We show that, very early after injury, wounds with impaired healing contain elevated levels of reactive oxygen and nitrogen species and, much like in humans, these levels increase with age. Moreover, the activity of anti-oxidant enzymes is not elevated, leading to buildup of oxidative stress in the wound environment. To induce chronicity, we exacerbated the redox imbalance by further inhibiting the antioxidant enzymes and by infecting the wounds with biofilm-forming bacteria isolated from the chronic wounds that developed naturally in these mice. These wounds do not re-epithelialize, the granulation tissue lacks vascularization and interstitial collagen fibers, they contain an antibiotic-resistant mixed bioflora with biofilm-forming capacity, and they stay open for several weeks. These findings are highly significant because they show for the first time that chronic wounds can be generated in an animal model effectively and consistently. The availability of such a model will significantly propel the field forward because it can be used to develop strategies to regain redox balance that may result in inhibition of biofilm formation and result in restoration of healthy wound tissue. Furthermore, the model can lead to the understanding of other fundamental mechanisms of chronic wound development that can potentially lead to novel therapies.
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- 2014
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41. Ceramide kinase is required for a normal eicosanoid response and the subsequent orderly migration of fibroblasts.
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Wijesinghe DS, Brentnall M, Mietla JA, Hoeferlin LA, Diegelmann RF, Boise LH, and Chalfant CE
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- Animals, Cell Movement genetics, Ceramides genetics, Ceramides metabolism, Fibroblasts cytology, Mice, Mice, Knockout, Phosphotransferases (Alcohol Group Acceptor) genetics, Wound Healing genetics, Cell Movement drug effects, Eicosanoids pharmacology, Fibroblasts metabolism, Phosphotransferases (Alcohol Group Acceptor) metabolism, Wound Healing drug effects
- Abstract
In these studies, the role of ceramide-1-phosphate (C1P) in the wound-healing process was investigated. Specifically, fibroblasts isolated from mice with the known anabolic enzyme for C1P, ceramide kinase (CERK), ablated (CERK(-/-) mice) and their wild-type littermates (CERK(+/+)) were subjected to in vitro wound-healing assays. Simulation of mechanical trauma of a wound by scratching a monolayer of fibroblasts from CERK(+/+) mice demonstrated steadily increasing levels of arachidonic acid in a time-dependent manner in stark contrast to CERK(-/-) fibroblasts. This observed difference was reflected in scratch-induced eicosanoid levels. Similar, but somewhat less intense, changes were observed in a more complex system utilizing skin biopsies obtained from CERK-null mice. Importantly, C1P levels increased during the early stages of human wound healing correlating with the transition from the inflammatory stage to the peak of the fibroplasia stage (e.g., proliferation and migration of fibroblasts). Finally, the loss of proper eicosanoid response translated into an abnormal migration pattern for the fibroblasts isolated from CERK(-/-) As the proper migration of fibroblasts is one of the necessary steps of wound healing, these studies demonstrate a novel requirement for the CERK-derived C1P in the proper healing response of wounds.
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- 2014
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42. Dietary fish oil improves endothelial function and lowers blood pressure via suppression of sphingolipid-mediated contractions in spontaneously hypertensive rats.
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van den Elsen LW, Spijkers LJ, van den Akker RF, van Winssen AM, Balvers M, Wijesinghe DS, Chalfant CE, Garssen J, Willemsen LE, Alewijnse AE, and Peters SL
- Subjects
- Animals, Chromatography, Liquid, Mass Spectrometry, Muscle Contraction drug effects, Rats, Rats, Inbred SHR, Sphingomyelin Phosphodiesterase metabolism, Thromboxane B2 blood, Blood Pressure drug effects, Dietary Fats, Unsaturated pharmacology, Endothelium, Vascular drug effects, Fish Oils pharmacology, Sphingolipids physiology
- Abstract
Objective: Long-chain n-3 polyunsaturated fatty acids from oily fish reduce blood pressure (BP) in hypertension. Previously, we demonstrated that hypertension is associated with marked alterations in sphingolipid biology and elevated ceramide-induced vasoconstriction. Here we investigated in spontaneously hypertensive rats (SHRs) whether fish oil improves endothelial function including reduced vascular contraction induced via the sphingolipid cascade, resulting in reduced BP., Methods: Twelve-week-old SHRs were fed a control or fish oil-enriched diet during 12 weeks, and BP was recorded. Plasma sphingolipid levels were quantified by mass spectrometry and the response of isolated carotid arteries towards different stimuli was measured. Furthermore, erythrocyte membrane fatty acid composition, thromboxane A2 formation and cytokine secretion in ex-vivo lipopolysaccharide-stimulated thoracic aorta segments were determined., Results: The fish oil diet reduced the mean arterial BP (P < 0.001) and improved endothelial function, as indicated by a substantially increased relaxation potential towards ex-vivo methacholine exposure of the carotid arteries (P < 0.001). The long-chain n-3 polyunsaturated fatty acid diet resulted in altered levels of specific (glucosyl)ceramide subspecies (P < 0.05), reduced membrane arachidonic acid content (P < 0.001) and decreased thromboxane concentrations in plasma (P < 0.01). Concomitantly, the fish oil diet largely reduced ceramide-induced contractions (P < 0.01), which are predominantly mediated by thromboxane. Furthermore, thromboxane A2 and interleukin-10 were reduced in supernatants of lipopolysaccharide-stimulated thoracic aorta of SHRs fed the fish oil diet while RANTES (regulated on activation, normal T-cell expressed and secreted) was enhanced. This may contribute to reduced vasoconstriction in vivo., Conclusions: Dietary fish oil lowers BP in SHRs and improves endothelial function in association with suppression of sphingolipid-dependent vascular contraction.
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- 2014
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43. Resolution of sterile inflammation: role for vitamin C.
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Mohammed BM, Fisher BJ, Huynh QK, Wijesinghe DS, Chalfant CE, Brophy DF, Fowler AA 3rd, and Natarajan R
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- Animals, Blotting, Western, Cell Line, Humans, Inflammation chemically induced, Inflammation metabolism, Lipopolysaccharides pharmacology, Macrophages drug effects, Macrophages metabolism, Macrophages, Peritoneal drug effects, Macrophages, Peritoneal metabolism, Mice, Mice, Knockout, Mice, Transgenic, Microscopy, Fluorescence, Peritonitis chemically induced, Peritonitis drug therapy, Peritonitis metabolism, Real-Time Polymerase Chain Reaction, Thioglycolates toxicity, Ascorbic Acid metabolism, Ascorbic Acid therapeutic use, Inflammation drug therapy
- Abstract
Introduction: Macrophage reprogramming is vital for resolution of acute inflammation. Parenteral vitamin C (VitC) attenuates proinflammatory states in murine and human sepsis. However information about the mechanism by which VitC regulates resolution of inflammation is limited., Methods: To examine whether physiological levels of VitC modulate resolution of inflammation, we used transgenic mice lacking L-gulono-γ-lactone oxidase. VitC sufficient/deficient mice were subjected to a thioglycollate-elicited peritonitis model of sterile inflammation. Some VitC deficient mice received daily parenteral VitC (200 mg/kg) for 3 or 5 days following thioglycollate infusion. Peritoneal macrophages harvested on day 3 or day 5 were examined for intracellular VitC levels, pro- and anti-inflammatory protein and lipid mediators, mitochondrial function, and response to lipopolysaccharide (LPS). The THP-1 cell line was used to determine the modulatory activities of VitC in activated human macrophages., Results: VitC deficiency significantly delayed resolution of inflammation and generated an exaggerated proinflammatory response to in vitro LPS stimulation. VitC sufficiency and in vivo VitC supplementation restored macrophage phenotype and function in VitC deficient mice. VitC loading of THP-1 macrophages attenuated LPS-induced proinflammatory responses., Conclusion: VitC sufficiency favorably modulates macrophage function. In vivo or in vitro VitC supplementation restores macrophage phenotype and function leading to timely resolution of inflammation.
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- 2014
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44. Systems-Level Lipid Analysis Methodologies for Qualitative and Quantitative Investigation of Lipid Signaling Events During Wound Healing.
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Wijesinghe DS and Chalfant CE
- Abstract
Objective: Accumulating evidence implicates a prominent role for lipid signaling molecules in the regulation of wound healing. These lipids regulate hemostasis, onset and resolution of inflammation, migration and proliferation cells, angiogenesis, epithelialization, and remodeling of collagen. The objective of this overview is to demonstrate the applicability of systems level lipid analyses to identify and quantify lipid involved in events leading to wound healing., Approach: Current advances in liquid chromatography coupled to tandem mass spectrometry have provided the means for carrying out quantitative and qualitative analysis of lipids at a systems level. This emerging field is collectively referred to as lipidomics and its potential in wound healing research is largely ignored., Results: While comprehensive applications of lipidomics in wound healing are limited, studies carried out by the authors as well as others demonstrate distinct changes in the lipidome during the wound healing process., Innovation: Until recently, investigations into lipids were limited to the study of a few lipids at a time. Lipidomics approaches provide the capability to quantitatively and qualitatively assay almost the full complement of lipid signaling circuits at the same time. This allows obtaining a system level understanding of changes to the entire lipidome during the wound healing process., Conclusion: The technology provides promising approach to understanding new signaling pathways based on lipids involved in wound healing. The understanding gained from such studies has the potential for the development of novel lipid based treatment strategies to promote wound healing.
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- 2013
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45. Non-vesicular trafficking by a ceramide-1-phosphate transfer protein regulates eicosanoids.
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Simanshu DK, Kamlekar RK, Wijesinghe DS, Zou X, Zhai X, Mishra SK, Molotkovsky JG, Malinina L, Hinchcliffe EH, Chalfant CE, Brown RE, and Patel DJ
- Subjects
- Animals, Apoproteins chemistry, Arachidonic Acid metabolism, Biological Transport, Carrier Proteins chemistry, Carrier Proteins genetics, Cell Membrane metabolism, Cell Nucleus metabolism, Ceramides chemistry, Crystallography, X-Ray, Cytosol metabolism, Humans, Hydrophobic and Hydrophilic Interactions, Mice, Models, Molecular, Phosphatidic Acids chemistry, Phosphatidic Acids metabolism, Phospholipid Transfer Proteins, Protein Conformation, Protein Folding, Substrate Specificity, trans-Golgi Network metabolism, Carrier Proteins metabolism, Ceramides metabolism, Eicosanoids metabolism
- Abstract
Phosphorylated sphingolipids ceramide-1-phosphate (C1P) and sphingosine-1-phosphate (S1P) have emerged as key regulators of cell growth, survival, migration and inflammation. C1P produced by ceramide kinase is an activator of group IVA cytosolic phospholipase A2α (cPLA2α), the rate-limiting releaser of arachidonic acid used for pro-inflammatory eicosanoid production, which contributes to disease pathogenesis in asthma or airway hyper-responsiveness, cancer, atherosclerosis and thrombosis. To modulate eicosanoid action and avoid the damaging effects of chronic inflammation, cells require efficient targeting, trafficking and presentation of C1P to specific cellular sites. Vesicular trafficking is likely but non-vesicular mechanisms for C1P sensing, transfer and presentation remain unexplored. Moreover, the molecular basis for selective recognition and binding among signalling lipids with phosphate headgroups, namely C1P, phosphatidic acid or their lyso-derivatives, remains unclear. Here, a ubiquitously expressed lipid transfer protein, human GLTPD1, named here CPTP, is shown to specifically transfer C1P between membranes. Crystal structures establish C1P binding through a novel surface-localized, phosphate headgroup recognition centre connected to an interior hydrophobic pocket that adaptively expands to ensheath differing-length lipid chains using a cleft-like gating mechanism. The two-layer, α-helically-dominated 'sandwich' topology identifies CPTP as the prototype for a new glycolipid transfer protein fold subfamily. CPTP resides in the cell cytosol but associates with the trans-Golgi network, nucleus and plasma membrane. RNA interference-induced CPTP depletion elevates C1P steady-state levels and alters Golgi cisternae stack morphology. The resulting C1P decrease in plasma membranes and increase in the Golgi complex stimulates cPLA2α release of arachidonic acid, triggering pro-inflammatory eicosanoid generation.
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- 2013
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46. Loss of miR-29b following acute ischemic stroke contributes to neural cell death and infarct size.
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Khanna S, Rink C, Ghoorkhanian R, Gnyawali S, Heigel M, Wijesinghe DS, Chalfant CE, Chan YC, Banerjee J, Huang Y, Roy S, and Sen CK
- Subjects
- Animals, Antioxidants pharmacology, Arachidonate 12-Lipoxygenase physiology, Blotting, Western, Cell Survival genetics, Cell Survival physiology, Cells, Cultured, Eicosanoids metabolism, Female, Fluorescent Antibody Technique, Glutathione metabolism, Magnetic Resonance Imaging, Membrane Potential, Mitochondrial drug effects, Mice, Mice, Inbred C57BL, MicroRNAs antagonists & inhibitors, Pregnancy, RNA biosynthesis, RNA isolation & purification, RNA, Small Interfering pharmacology, Rats, Rats, Sprague-Dawley, Real-Time Polymerase Chain Reaction, Tocotrienols pharmacology, Transfection, Brain Ischemia genetics, Brain Ischemia pathology, Cell Death genetics, Cell Death physiology, Cerebral Infarction genetics, Cerebral Infarction pathology, MicroRNAs genetics, MicroRNAs physiology, Neurons pathology, Stroke genetics, Stroke pathology
- Abstract
Glutathione depletion and 12-lipoxygenase-dependent metabolism of arachidonic acid are known to be implicated in neurodegeneration associated with acute ischemic stroke. The objective of this study was to investigate the significance of miR-29 in neurodegeneration associated with acute ischemic stroke. Neural cell death caused by arachidonic acid insult of glutathione-deficient cells was preceded by a 12-lipoxygenase-dependent loss of miR-29b. Delivery of miR-29b mimic to blunt such loss was neuroprotective. miR-29b inhibition potentiated such neural cell death. 12-Lipoxygenase knockdown and inhibitors attenuated the loss of miR-29b in challenged cells. In vivo, stroke caused by middle-cerebral artery occlusion was followed by higher 12-lipoxygenase activity and loss of miR-29b as detected in laser-captured infarct site tissue. 12-Lipoxygenase knockout mice demonstrated protection against such miR loss. miR-29b gene delivery markedly attenuated stroke-induced brain lesion. Oral supplementation of α-tocotrienol, a vitamin E 12-lipoxygenase inhibitor, rescued stroke-induced loss of miR-29b and minimized lesion size. This work provides the first evidence demonstrating that loss of miR-29b at the infarct site is a key contributor to stroke lesion. Such loss is contributed by activity of the 12-lipoxygenase pathway providing maiden evidence linking arachidonic acid metabolism to miR-dependent mechanisms in stroke.
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- 2013
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47. Characterization of eicosanoid synthesis in a genetic ablation model of ceramide kinase.
- Author
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Mietla JA, Wijesinghe DS, Hoeferlin LA, Shultz MD, Natarajan R, Fowler AA 3rd, and Chalfant CE
- Subjects
- Animals, Cells, Cultured, Ceramides blood, Female, Mice, Mice, Inbred BALB C, Mice, Knockout, Phosphotransferases (Alcohol Group Acceptor) genetics, Phosphotransferases (Alcohol Group Acceptor) metabolism, Pregnancy, Disease Models, Animal, Eicosanoids biosynthesis, Phosphotransferases (Alcohol Group Acceptor) deficiency, Pregnancy, Animal
- Abstract
Multiple reports have demonstrated a role for ceramide kinase (CERK) in the production of eicosanoids. To examine the effects of the genetic ablation of CERK on eicosanoid synthesis, primary mouse embryonic fibroblasts (MEFs) and macrophages were isolated from CERK(-/-) and CERK(+/+) mice, and the ceramide-1-phosphate (C1P) and eicosanoid profiles were investigated. Significant decreases were observed in multiple C1P subspecies in CERK-/- cells as compared to CERK(+/+) cells with overall 24% and 48% decreases in total C1P. In baseline experiments, the levels of multiple eicosanoids were significantly lower in the CERK(-/-) cells compared with wild-type cells. Importantly, induction of eicosanoid synthesis by calcium ionophore was significantly reduced in the CERK(-/-) MEFs. Our studies also demonstrate that the CERK(-/-) mouse has adapted to loss of CERK in regards to airway hyper-responsiveness as compared with CERK siRNA treatment. Overall, we demonstrate that there are significant differences in eicosanoid levels in ex vivo CERK(-/-) cells compared with wild-type counterparts, but the effect of the genetic ablation of CERK on eicosanoid synthesis and the serum levels of C1P was not apparent in vivo.
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- 2013
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48. Structural Stereochemistry of Androstene Hormones Determines Interactions with Human Androgen, Estrogen, and Glucocorticoid Receptors.
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Shaak TL, Wijesinghe DS, Chalfant CE, Diegelmann RF, Ward KR, and Loria RM
- Abstract
DHEA, 17 α -AED, 17 β -AED, and 17 β -AET exhibit strong biological activity that has been attributed to androgenic, estrogenic, or antiglucocorticoid activity in vivo and in vitro. This study compared DHEA, 17 α -AED, 17 β -AED, and 17 β -AET for their ability to activate the human AR, ER, and GR and determine the relative androgenicity, estrogenicity, and glucocorticoid activity. The results show that, at the receptor level, these androstene hormones are weak AR and even weaker ER activators. Direct androstene hormone activation of the human AR, ER α , and ER β may not be essential for their biological function. Similarly, these hormones indirectly activated the human GR, only in the presence of high dexamethasone concentrations. These results underscore the major difference between androstene hormone interactions with these nuclear receptors and their biological effects.
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- 2013
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49. The role of ceramide-1-phosphate in biological functions.
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Hoeferlin LA, Wijesinghe DS, and Chalfant CE
- Subjects
- Animals, Cell Proliferation, Cell Survival, Group IV Phospholipases A2 physiology, Humans, Phosphotransferases (Alcohol Group Acceptor) physiology, Ceramides physiology
- Abstract
In mammalian cells, cermide-1-phosphate (C1P) is produced via the ATP-dependent mechanism of converting ceramide to C1P by the enzyme, ceramide kinase (CERK). CERK was first described as a calcium-stimulated lipid kinase that co-purified with brain synaptic vesicles, and to date, CERK is the only identified mammalian enzyme known to produce C1P in cells. C1P has steadily emerged as a bioactive sphingolipid involved in cell proliferation, macrophage migration, and inflammatory events. The recent generation of the CERK knockout mouse and the development of CERK inhibitors have furthered our current understanding of CERK-derived C1P in regulating biological processes. In this chapter, the history of C1P as well as the biological functions attributed to C1P are reviewed.
- Published
- 2013
- Full Text
- View/download PDF
50. Metabolic gene remodeling and mitochondrial dysfunction in failing right ventricular hypertrophy secondary to pulmonary arterial hypertension.
- Author
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Gomez-Arroyo J, Mizuno S, Szczepanek K, Van Tassell B, Natarajan R, dos Remedios CG, Drake JI, Farkas L, Kraskauskas D, Wijesinghe DS, Chalfant CE, Bigbee J, Abbate A, Lesnefsky EJ, Bogaard HJ, and Voelkel NF
- Subjects
- Animals, Disease Models, Animal, Familial Primary Pulmonary Hypertension, Heart Failure etiology, Heart Failure physiopathology, Hypertension, Pulmonary complications, Hypertension, Pulmonary metabolism, Hypertrophy, Right Ventricular complications, Hypertrophy, Right Ventricular metabolism, Male, Mitochondria, Heart metabolism, Oxidation-Reduction, Rats, Rats, Sprague-Dawley, Heart Failure genetics, Hypertension, Pulmonary genetics, Hypertrophy, Right Ventricular genetics, Mitochondria, Heart genetics, Mitochondrial Turnover genetics, Myocytes, Cardiac metabolism, Ventricular Remodeling genetics
- Abstract
Background: Right ventricular (RV) dysfunction (RVD) is the most frequent cause of death in patients with pulmonary arterial hypertension. Although abnormal energy substrate use has been implicated in the development of chronic left heart failure, data describing such metabolic remodeling in RVD remain incomplete. Thus, we sought to characterize metabolic gene expression changes and mitochondrial dysfunction in functional and dysfunctional RV hypertrophy., Methods and Results: Two different rat models of RV hypertrophy were studied. The model of RVD (SU5416/hypoxia) exhibited a significantly decreased gene expression of peroxisome proliferator-activated receptor-γ coactivator-1α, peroxisome proliferator-activated receptor-α and estrogen-related receptor-α. The expression of multiple peroxisome proliferator-activated receptor-γ coactivator-1α target genes required for fatty acid oxidation was similarly decreased. Decreased peroxisome proliferator-activated receptor-γ coactivator-1α expression was also associated with a net loss of mitochondrial protein and oxidative capacity. Reduced mitochondrial number was associated with a downregulation of transcription factor A, mitochondrial, and other genes required for mitochondrial biogenesis. Electron microscopy demonstrated that, in RVD tissue, mitochondria had abnormal shape and size. Lastly, respirometric analysis demonstrated that mitochondria isolated from RVD tissue had a significantly reduced ADP-stimulated (state 3) rate for complex I. Conversely, functional RV hypertrophy in the pulmonary artery banding model showed normal expression of peroxisome proliferator-activated receptor-γ coactivator-1α, whereas the expression of fatty acid oxidation genes was either preserved or unregulated. Moreover, pulmonary artery banding-RV tissue exhibited preserved transcription factor A mitochondrial expression and mitochondrial respiration despite elevated RV pressure-overload., Conclusions: Right ventricular dysfunction, but not functional RV hypertrophy in rats, demonstrates a gene expression profile compatible with a multilevel impairment of fatty acid metabolism and significant mitochondrial dysfunction, partially independent of chronic pressure-overload.
- Published
- 2013
- Full Text
- View/download PDF
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