RESUMO
BACKGROUND: This work aims to present a fast, affordable, and reproducible three-cell co-culture system that could represent the different cellular mechanisms of atherosclerosis, extending from atherogenesis to pathological intimal thickening. METHODS AND RESULTS: We built four culture models: (i) Culture model #1 (representing normal arterial intima), where human coronary artery endothelial cells were added on top of Matrigel-coated collagen type I matrix, (ii) Culture model #2 (representing atherogenesis), which demonstrated the subendothelial accumulation and oxidative modification of low-density lipoproteins (LDL), (iii) Culture model #3 (representing intimal xanthomas), which demonstrated the monocyte adhesion to the endothelial cell monolayer, transmigration into the subendothelial space, and transformation to lipid-laden macrophages, (iv) Culture model #4 (representing pathological intimal thickening), which incorporated multiple layers of human coronary artery smooth muscle cells within the matrix. Coupling this model with different shear stress conditions revealed the effect of low shear stress on the oxidative modification of LDL and the upregulation of pro-inflammatory molecules and matrix-degrading enzymes. Using electron microscopy, immunofluorescence confocal microscopy, protein and mRNA quantification assays, we showed that the behaviors exhibited by the endothelial cells, macrophages and vascular smooth muscle cells in these models were very similar to those exhibited by these cell types in nascent and intermediate atherosclerotic plaques in humans. The preparation time of the cultures was 24 hours. CONCLUSION: We present three-cell co-culture models of human atherosclerosis. These models have the potential to allow cost- and time-effective investigations of the mechanobiology of atherosclerosis and new anti-atherosclerotic drug therapies.
Assuntos
Aterosclerose , Células Endoteliais , Humanos , Técnicas de Cocultura , Células Endoteliais/metabolismo , Músculo Liso Vascular/metabolismo , Aterosclerose/metabolismo , Macrófagos/metabolismo , Miócitos de Músculo Liso/metabolismoRESUMO
Chronic inflammation plays a critical role in the pathogenesis of atherosclerosis. Currently, the mechanism(s) by which inflammation contributes to this disease are not entirely understood. Inflammation is known to induce oxidative stress, which can lead to lipid peroxidation. Lipid peroxidation can result in the production of reactive by-products that can oxidatively modify macromolecules including DNA, proteins, and lipoproteins. A major reactive by-product of lipid peroxidation is malondialdehyde (MDA). MDA can subsequently break down to form acetaldehyde (AA). These two aldehydes can covalently interact with the epsilon (ε)-amino group of lysines within proteins and lipoproteins leading to the formation of extremely stable, highly immunogenic malondialdehyde/acetaldehyde adducts (MAA-adducts). The aim of this study was to investigate the inflammatory response to MAA-modified human serum albumin (HSA-MAA) and low-density lipoprotein (LDL-MAA). We found that animals injected with LDL-MAA generate antibodies specific to MAA-adducts. The level of anti-MAA antibodies were further increased in an animal model of atherosclerosis fed a Western diet. An animal model that combined both high fat diet and immunization of MAA-modified protein resulted in a dramatic increase in antibodies to MAA-adducts and vascular fat accumulation compared with controls. In vitro exposure of endothelial cells and macrophages to MAA-modified proteins resulted in increased fat accumulation as well as increased expression of adhesion molecules and pro-inflammatory cytokines. The expression of cytokines varied between the different cell lines and was unique to the individual modified proteins. The results of these studies demonstrate that different MAA-modified proteins elicit unique responses in different cell types. Additionally, the presence of MAA-modified proteins appears to modulate cellular metabolism leading to increased accumulation of triglycerides and further progression of the inflammatory response.
Assuntos
Inflamação/metabolismo , Lipoproteínas LDL/imunologia , Lipoproteínas LDL/metabolismo , Processamento de Proteína Pós-Traducional , Albumina Sérica Humana/imunologia , Albumina Sérica Humana/metabolismo , Acetaldeído/metabolismo , Animais , Aterosclerose/etiologia , Aterosclerose/imunologia , Aterosclerose/metabolismo , Doenças Cardiovasculares/etiologia , Doenças Cardiovasculares/imunologia , Doenças Cardiovasculares/metabolismo , Moléculas de Adesão Celular/metabolismo , Células Cultivadas , Citocinas/metabolismo , Modelos Animais de Doenças , Células Endoteliais/imunologia , Células Endoteliais/metabolismo , Feminino , Humanos , Inflamação/etiologia , Inflamação/imunologia , Metabolismo dos Lipídeos/imunologia , Macrófagos/imunologia , Macrófagos/metabolismo , Masculino , Malondialdeído/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Ratos , Ratos Sprague-DawleyAssuntos
Células Epiteliais Alveolares/metabolismo , COVID-19/metabolismo , Células Endoteliais/metabolismo , Fibroblastos/metabolismo , Interleucina-33/metabolismo , Pulmão/metabolismo , Miócitos de Músculo Liso/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Estudos de Casos e Controles , Feminino , Humanos , Fibrose Pulmonar Idiopática/metabolismo , Masculino , Pessoa de Meia-Idade , Doença Pulmonar Obstrutiva Crônica/metabolismo , Proteína C Associada a Surfactante Pulmonar/metabolismo , SARS-CoV-2 , Vimentina/metabolismo , Adulto JovemRESUMO
PURPOSE: To determine physician radiation exposure when using partial-angle computed tomography (CT) fluoroscopy (PACT) vs conventional full-rotation CT and whether there is an optimal tube/detector position at which physician dose is minimized. MATERIALS AND METHODS: Physician radiation dose (entrance air kerma) was measured for full-rotation CT (360°) and PACT (240°) at all tube/detector positions using a human-mimicking phantom placed in a 64-channel multidetector CT. Parameters included 120 kV, 20- and 40-mm collimation, and 100 mA. The mean, standard deviation, and increase/decrease in physician dose compared with a full-rotation scan were reported. RESULTS: Physician radiation exposure during CT fluoroscopy with PACT was highly dependent on the position of the tube/detector during scanning. The lowest PACT physician dose was when the physician was on the detector side (center view angle 116°; -35% decreased dose vs full-angle CT). The highest PACT physician dose was with the physician on the tube side (center view angle 298°; +34% increased dose vs full-angle CT), all doses P <.05 vs full-rotation CT. CONCLUSIONS: Partial-angle CT has the potential to both significantly increase or decrease physician radiation dose during CT fluoroscopy-guided procedures. The detector/tube position has a profound effect on physician dose. The lowest dose during PACT was achieved when the physician was located on the detector side (ie, distant from the tube). This data could be used to optimize CT fluoroscopy parameters to reduce physician radiation exposure for PACT-capable scanners.
Assuntos
Tomografia Computadorizada Multidetectores , Exposição Ocupacional , Doses de Radiação , Exposição à Radiação , Radiografia Intervencionista , Radiologistas , Fluoroscopia , Humanos , Tomografia Computadorizada Multidetectores/efeitos adversos , Tomografia Computadorizada Multidetectores/instrumentação , Exposição Ocupacional/efeitos adversos , Exposição Ocupacional/prevenção & controle , Saúde Ocupacional , Imagens de Fantasmas , Exposição à Radiação/efeitos adversos , Exposição à Radiação/prevenção & controle , Radiografia Intervencionista/efeitos adversos , Radiografia Intervencionista/instrumentação , Medição de Risco , Fatores de Risco , Tomógrafos ComputadorizadosRESUMO
Increased matrix metalloprotease 9 (MMP9) after myocardial infarction (MI) exacerbates ischemia-induced chronic heart failure (CHF). Autophagy is cardioprotective during CHF; however, whether increased MMP9 suppresses autophagic activity in CHF is unknown. This study aimed to determine whether increased MMP9 suppressed autophagic flux and MMP9 inhibition increased autophagic flux in the heart of rats with post-MI CHF. Sprague-Dawley rats underwent either sham surgery or coronary artery ligation 6-8 wk before being treated with MMP9 inhibitor for 7 days, followed by cardiac autophagic flux measurement with lysosomal inhibitor bafilomycin A1. Furthermore, autophagic flux was measured in vitro by treating H9c2 cardiomyocytes with two independent pharmacological MMP9 inhibitors, salvianolic acid B (SalB) and MMP9 inhibitor-I, and CRISPR/cas9-mediated MMP9 genetic ablation. CHF rats showed cardiac infarct, significantly increased left ventricular end-diastolic pressure (LVEDP), and increased MMP9 activity and fibrosis in the peri-infarct areas of left ventricular myocardium. Measurement of the autophagic markers LC3B-II and p62 with lysosomal inhibition showed decreased autophagic flux in the peri-infarct myocardium. Treatment with SalB for 7 days in CHF rats decreased MMP9 activity and cardiac fibrosis but increased autophagic flux in the peri-infarct myocardium. As an in vitro corollary study, measurement of autophagic flux in H9c2 cardiomyocytes and fibroblasts showed that pharmacological inhibition or genetic ablation of MMP9 upregulates autophagic flux. These data are consistent with our observations that MMP9 inhibition upregulates autophagic flux in the heart of rats with CHF. In conclusion, the results in this study suggest that the beneficial outcome of MMP9 inhibition in pathological cardiac remodeling is in part mediated by improved autophagic flux.NEW & NOTEWORTHY This study elucidates that the improved cardiac extracellular matrix (ECM) remodeling and cardioprotective effect of matrix metalloprotease 9 (MMP9) inhibition in chronic heart failure (CHF) are via increased autophagic flux. Autophagy is cardioprotective; however, the mechanism of autophagy suppression in CHF is unknown. We for the first time demonstrated here that increased MMP9 suppressed cardiac autophagy and ablation of MMP9 increased cardiac autophagic flux in CHF rats. Restoring the physiological level of autophagy in the failing heart is a challenge, and our study addressed this challenge. The novelty and highlights of this report are as follows: 1) MMP9 regulates cardiomyocyte and fibroblast autophagy, 2) MMP9 inhibition protects CHF after myocardial infarction (MI) via increased cardiac autophagic flux, 3) MMP9 inhibition increased cardiac autophagy via activation of AMP-activated protein kinase (AMPK)α, Beclin-1, Atg7 pathway and suppressed mechanistic target of rapamycin (mTOR) pathway.
Assuntos
Autofagia/efeitos dos fármacos , Benzofuranos/farmacologia , Fibroblastos/efeitos dos fármacos , Insuficiência Cardíaca/tratamento farmacológico , Metaloproteinase 9 da Matriz/metabolismo , Inibidores de Metaloproteinases de Matriz/farmacologia , Miócitos Cardíacos/efeitos dos fármacos , Animais , Linhagem Celular , Modelos Animais de Doenças , Fibroblastos/enzimologia , Fibroblastos/patologia , Fibrose , Insuficiência Cardíaca/enzimologia , Insuficiência Cardíaca/patologia , Insuficiência Cardíaca/fisiopatologia , Masculino , Metaloproteinase 9 da Matriz/genética , Camundongos , Miócitos Cardíacos/enzimologia , Miócitos Cardíacos/patologia , Ratos Sprague-Dawley , Transdução de Sinais , Função Ventricular Esquerda/efeitos dos fármacos , Remodelação Ventricular/efeitos dos fármacosRESUMO
Many medications exhibit clinical benefits that are unrelated to their primary therapeutic uses. In many cases, the mechanisms underpinning these pleotropic effects are unknown. Two commonly prescribed medications that exhibit pleotropic benefits in cardiovascular disease and other diseases associated with chronic inflammation are methotrexate (MTX) and doxycycline (DOX). The vast majority of cardiovascular disease is associated with atherosclerosis. Because atherosclerosis is a chronic inflammatory disease, possible mechanisms by which MTX and DOX reduce inflammation have been investigated. Interestingly, the primary structure of both of these medications contain aromatic phenolic rings, which resemble polyphenols that are known to possess antioxidant activity. Inflammation and oxidative stress are intimately related. Inflammation promotes oxidative stress, which in turn leads to further inflammation; in this way, oxidative stress and inflammation can establish a self-perpetuating cycle. It has been shown that MTX and DOX act as antioxidants and are capable of scavenging free radicals and the reactive oxygen species (ROS) superoxide (O2-). Furthermore, both MTX and DOX inhibit the formation of malondialdehyde acetaldehyde (MAA) adducts, products of oxidative stress and lipid peroxidation. Importantly, MAA-adducts are highly immunogenic and initiate inflammatory responses; thereby, fueling the cycle of inflammation and oxidative stress that results in chronic inflammation. Thus, reducing the formation of MAA-adducts may ameliorate inflammation that leads to ROS production and in this way, break the self-sustaining cycle of oxidative stress and inflammation. It is possible that the under-recognized antioxidant properties of these medications may be a mechanism by which they and other medications provide pleotropic benefit in the treatment of chronic inflammatory disease.
Assuntos
Antioxidantes/farmacologia , Doxiciclina/farmacologia , Metotrexato/farmacologia , Animais , Aterosclerose/tratamento farmacológico , Aterosclerose/fisiopatologia , Doenças Cardiovasculares/tratamento farmacológico , Doenças Cardiovasculares/fisiopatologia , Humanos , Inflamação/tratamento farmacológico , Inflamação/fisiopatologia , Peroxidação de Lipídeos/efeitos dos fármacos , Malondialdeído/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismoRESUMO
Patient dose from 2.5 MV images on the TrueBeam linear accelerator is not easily quantified, primarily because this beam energy is not normally modeled by commercial treatment planning systems. In this work we present the feasibility of using the Eclipse® treatment planning system to model this beam. The Acuros XB and the AAA dose calculation algorithms were tested. Profiles, PDDs, and output factors were measured for the 2.5 MV unflattened imaging beam and used for beam modeling. The algorithms were subsequently verified using MPPG 5.a guidelines. Calculated doses with both algorithms agreed with the measurement data to within the following criteria recommended for conventional therapeutic MV beams: 2% local dose-difference in the high-dose region, 3% global difference in the low-dose region, 3 mm distance to agreement in the penumbra, and a gamma pass rate of >95% for 3%/3 mm criteria. Acuros was able to accurately calculate dose through cork and bone-equivalent heterogeneities. AAA was able to accurately calculate dose through the bone-equivalent heterogeneity but did not pass within the recommended criteria for the cork heterogeneity. For the 2.5 MV imaging beam, both the AAA and Acuros algorithms provide calculated doses that agree with measured results well within the 20% criteria for imaging beams recommended by AAPM TG-180.
Assuntos
Algoritmos , Neoplasias/radioterapia , Aceleradores de Partículas/instrumentação , Planejamento de Assistência ao Paciente/normas , Imagens de Fantasmas , Fótons/uso terapêutico , Planejamento da Radioterapia Assistida por Computador/métodos , Humanos , Órgãos em Risco/efeitos da radiação , Dosagem RadioterapêuticaRESUMO
OBJECTIVE: To compare anti-malondialdehyde-acetaldehyde (MAA) antibody concentrations between rheumatoid arthritis (RA) patients and healthy and rheumatic disease controls. METHODS: Anti-MAA antibody (IgA, IgM, IgG) was measured using ELISA and banked serum from patients with RA (nâ¯=â¯284), osteoarthritis (OA, nâ¯=â¯330), spondyloarthropathy (SpA, nâ¯=â¯50), and systemic lupus erythematosus (SLE, nâ¯=â¯88) as well as healthy controls (nâ¯=â¯82). Anti-MAA antibody concentrations and the frequency of positivity were compared across groups. Multivariable linear regression analysis limited to RA and OA patients (due to sample size and data availability) was used to identify factors associated with anti-MAA antibody concentrations. RESULTS: Although RA patients demonstrated among the highest circulating concentrations across isotypes, only IgA anti-MAA antibody was significantly higher than all other groups (pâ¯≤â¯0.02). Proportions (7% to 74%) of OA and SLE (less so for SpA) samples were positive for anti-MAA antibody, limiting the discriminatory capacity of anti-MAA antibody in RA (positive in 18% to 80%). In analyses limited to those with RA or OA, factors associated with higher anti-MAA antibody concentrations included RA case status, younger age (IgM), male sex (IgG), African American race (IgA, IgG) and current smoking (IgA). C-reactive protein levels and comorbidities were not associated with anti-MAA antibody concentrations. CONCLUSION: With the possible exception of the IgA isotype, serum anti-MAA antibodies measured with currently available assays do not appear to adequately discriminate RA from other rheumatic conditions. With the identification of specific proteins that are MAA-modified in diseased tissues and requisite assay refinement, anti-MAA antibody holds potential promise as a biomarker in RA.
Assuntos
Artrite Reumatoide/imunologia , Autoanticorpos/sangue , Doenças Reumáticas/imunologia , Acetaldeído/imunologia , Adulto , Idoso , Artrite Reumatoide/diagnóstico , Estudos de Casos e Controles , Diagnóstico Diferencial , Feminino , Humanos , Imunidade Humoral , Masculino , Malondialdeído/imunologia , Pessoa de Meia-Idade , Doenças Reumáticas/diagnóstico , Fatores de RiscoRESUMO
Precision-cut liver slices (PCLSs) provide a novel model for studies of alcoholic liver disease (ALD). This is relevant, as in vivo ethanol exposure does not appear to generate significant liver damage in ethanol-fed mice, except in the National Institute on Alcohol Abuse and Alcoholism binge model of ALD. Previous studies have shown that the two metabolites of ethanol consumption, malondialdhyde (MDA) and acetaldehyde (AA), combine to form MDA-AA (MAA) adducts, which have been correlated with the development and progression of ALD. In this study, murine PCLSs were incubated with ethanol and examined for the production of MAA adducts. PCLSs were homogenized, and homogenates were injected into C57BL/6 mice. PCLSs from control-, pair-, and ethanol-fed animals served as targets in in situ cytotoxic assays using primed T cells from mice hyperimmunized with control or ethanol-exposed PCLS homogenates. A CD45.1/CD45.2 passive-transfer model was used to determine whether T cells from the spleens of mice hyperimmunized with PCLS ethanol-exposed homogenates trafficked to the liver. PCLSs incubated with ethanol generated MAA-modified proteins in situ. Cytotoxic (CD8+) T cells from immunized mice killed naïve PCLSs from control- and pair-fed mice in vitro, a response that was blunted in PCLSs from ethanol-fed mice. Furthermore, CD45.1 CD8+ T cells from hyperimmunized mice trafficked to the liver but did not initiate liver damage. This study demonstrates that exposure to liver tissue damaged by ethanol mediates robust immune responses to well-characterized alcohol metabolites and native liver proteins in vitro. Moreover, although these proinflammatory T cells traffic to the liver, these responses appear to be dampened in vivo by locally acting pathways. NEW & NOTEWORTHY This study shows that the metabolites of ethanol and lipid breakdown produce malondialdehyde-acetaldehyde adducts in the precision-cut liver slice model system. Additionally, precision-cut liver slices exposed to ethanol and harboring malondialdehyde-acetaldehyde adducts generate liver-specific antibody and T cell responses in the spleens of naïve mice that could traffic to the liver.
Assuntos
Acetaldeído/imunologia , Autoimunidade , Fígado Gorduroso Alcoólico/imunologia , Hepatopatias Alcoólicas/imunologia , Fígado/imunologia , Malondialdeído/imunologia , Linfócitos T Citotóxicos/imunologia , Acetaldeído/metabolismo , Transferência Adotiva , Animais , Movimento Celular , Células Cultivadas , Modelos Animais de Doenças , Fígado Gorduroso Alcoólico/metabolismo , Feminino , Humanos , Técnicas In Vitro , Interleucina-6/imunologia , Interleucina-6/metabolismo , Antígenos Comuns de Leucócito/imunologia , Antígenos Comuns de Leucócito/metabolismo , Fígado/metabolismo , Hepatopatias Alcoólicas/metabolismo , Ativação Linfocitária , Malondialdeído/metabolismo , Camundongos Endogâmicos C57BL , Fenótipo , Baço/imunologia , Baço/metabolismo , Linfócitos T Citotóxicos/metabolismo , Linfócitos T Citotóxicos/transplanteRESUMO
OBJECTIVE: Abdominal aortic aneurysm (AAA) is a pathologic dilation of the aorta. Inflammation of the aortic wall has been shown to be involved in AAA formation. Malondialdehyde-acetaldehyde (MAA) adducts are MAA/protein hybrids with immunogenic, proinflammatory, and profibrotic properties. Levels of MAA adducts are elevated in patients with coronary artery disease; however, the role of MAA adducts in AAA is unclear. We hypothesize that levels of circulating antibodies against MAA adducts are increased in patients with AAA. METHODS: Plasma samples were collected from mice and patients with AAA and control patients with atherosclerosis but not AAA. AAA was induced in mice by a standard CaCl2 protocol, with matching sham mice. Plasma levels of anti-MAA antibodies were quantified by enzyme-linked immunosorbent assay. RESULTS: Patients with AAA exhibited higher levels of immunoglobulin G and immunoglobulin A anti-MAA antibody subtypes (P = .049 and .026, respectively) compared with control patients. Conversely, immunoglobulin M anti-MAA antibodies in AAA patients were lower compared with control patients (P = .018). In CaCl2-treated mice, immunoglobulin G anti-MAA antibodies were elevated after AAA formation (P = .006). CONCLUSIONS: The pattern of anti-MAA antibodies is able to distinguish between patients with AAA and patients with atherosclerosis but no AAA. These results demonstrate that MAA adducts are associated with AAA and suggest that they may play a role in either initiating or propagating chronic inflammation in AAA.
Assuntos
Acetaldeído/imunologia , Aneurisma da Aorta Abdominal/diagnóstico , Imunoglobulina A/sangue , Imunoglobulina G/sangue , Malondialdeído/imunologia , Acetaldeído/análogos & derivados , Idoso , Idoso de 80 Anos ou mais , Animais , Aneurisma da Aorta Abdominal/sangue , Aneurisma da Aorta Abdominal/induzido quimicamente , Aneurisma da Aorta Abdominal/imunologia , Biomarcadores/sangue , Cloreto de Cálcio , Estudos de Casos e Controles , Modelos Animais de Doenças , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Masculino , Malondialdeído/análogos & derivados , Camundongos Endogâmicos C57BL , Valor Preditivo dos Testes , Regulação para CimaRESUMO
OBJECTIVE: To examine whether vascular calcifications on hand films in RA might aid in determining mortality risk. METHODS: Hand radiographs from 906 RA patients were scored as positive or negative for vascular calcifications. Patient characteristics associated with vascular calcifications were assessed using multivariable logistic regression, and associations with mortality were examined using Cox proportional hazards regression. Cytokines and multiplex ACPA were measured in both groups. RESULTS: A total of 99 patients (11%) demonstrated radiographic vascular calcifications. Factors independently associated with vascular calcifications included diabetes [odds ratio (OR) 2.85; 95% CI 1.43, 5.66], cardiovascular disease at enrolment (OR 2.48; 95% CI 1.01, 6.09), prednisone use (OR 1.90; 95% CI 1.25, 2.91), current smoking (OR 0.06; 95% CI 0.01, 0.23) and former smoking (OR 0.36; 95% CI 0.27, 0.48) vs never smoking. In cytokine and ACPA subtype analysis, IL-4 and anti-citrullinated apolipoprotein E were significantly increased in patients with vascular calcifications in fully adjusted multivariable models. After multivariable adjustment, vascular calcifications were associated with an increase in all-cause mortality (hazard ratio 1.41; 95% CI 1.12, 1.78; P = 0.004). CONCLUSION: Vascular calcifications on hand radiographs were independently associated with increased all-cause mortality in RA. Mechanisms underpinning the associations of IL-4 and select ACPA with vascular calcifications and their utility as biomarkers predictive of cardiovascular disease risk in RA merit further study.
Assuntos
Artrite Reumatoide/diagnóstico por imagem , Autoanticorpos/sangue , Doenças Cardiovasculares/epidemiologia , Doenças Cardiovasculares/mortalidade , Mãos/diagnóstico por imagem , Calcificação Vascular/complicações , Calcificação Vascular/diagnóstico por imagem , Idoso , Idoso de 80 Anos ou mais , Apolipoproteínas E/sangue , Artrite Reumatoide/sangue , Artrite Reumatoide/imunologia , Biomarcadores/sangue , Feminino , Mãos/irrigação sanguínea , Humanos , Interleucina-4/sangue , Modelos Logísticos , Estudos Longitudinais , Masculino , Pessoa de Meia-Idade , Peptídeos Cíclicos/imunologia , Radiografia , Fatores de Risco , Taxa de SobrevidaRESUMO
A method, based on well-established trauma principles, is described for surgical management of serious intrathoracic bleeding complications that can occur during the extraction of pacemaker or defibrillator leads. Using this method, four patients who experienced rapid hemodynamic deterioration due to traumatic injury of the superior vena cava and its tributaries during defibrillator lead extraction underwent successful surgical repair. Perioperative preparation for high-risk lead extractions, management of major bleeding complications, and surgical repair techniques are discussed. Major bleeding complications can be managed effectively with this strategy leading to excellent overall success rates for extractions without mortality.
Assuntos
Desfibriladores Implantáveis/efeitos adversos , Remoção de Dispositivo/efeitos adversos , Hemorragia/etiologia , Hemorragia/cirurgia , Complicações Intraoperatórias/etiologia , Complicações Intraoperatórias/cirurgia , Marca-Passo Artificial/efeitos adversos , Assistência Perioperatória , Doenças Torácicas/etiologia , Doenças Torácicas/cirurgia , Procedimentos Cirúrgicos Torácicos/métodos , Adulto , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Risco , Resultado do Tratamento , Veia Cava Superior/lesões , Veia Cava Superior/cirurgiaRESUMO
Malondialdehyde-acetaldehyde adducts (MAA) have been implicated in atherosclerosis. The purpose of this study was to investigate the role of MAA in atherosclerotic disease. Serum samples from controls (n = 82) and patients with; non-obstructive coronary artery disease (CAD), (n = 40), acute myocardial infarction (AMI) (n = 42), or coronary artery bypass graft (CABG) surgery due to obstructive multi-vessel CAD (n = 72), were collected and tested for antibody isotypes to MAA-modifed human serum albumin (MAA-HSA). CAD patients had elevated relative levels of IgG and IgA anti-MAA, compared to control patients (p<0.001). AMI patients had a significantly increased relative levels of circulating IgG anti-MAA-HSA antibodies as compared to stable angina (p<0.03) or CABG patients (p<0.003). CABG patients had significantly increased relative levels of circulating IgA anti-MAA-HSA antibodies as compared to non-obstructive CAD (p<0.001) and AMI patients (p<0.001). Additionally, MAA-modified proteins were detected in the tissue of human AMI lesions. In conclusion, the IgM, IgG and IgA anti-MAA-HSA antibody isotypes are differentially and significantly associated with non-obstructive CAD, AMI, or obstructive multi-vessel CAD and may serve as biomarkers of atherosclerotic disease.
Assuntos
Acetaldeído/imunologia , Autoanticorpos/sangue , Doença da Artéria Coronariana/imunologia , Lipoproteínas LDL/imunologia , Malondialdeído/imunologia , Idoso , Biomarcadores/sangue , Estudos de Casos e Controles , Doença da Artéria Coronariana/sangue , Doença da Artéria Coronariana/diagnóstico , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Placa Aterosclerótica/sangue , Placa Aterosclerótica/imunologiaRESUMO
Oxidative stress from fat accumulation in the liver has many deleterious effects. Many believe that there is a second hit that causes relatively benign fat accumulation to transform into liver failure. Therefore, we evaluated the effects of ethanol on ex vivo precision-cut liver slice cultures (PCLS) from rats fed a high-fat diet resulting in fatty liver. Age-matched male Sprague-Dawley rats were fed either high-fat (obese) (45% calories from fat, 4.73 kcal/g) or control diet for 13 mo. PCLS were prepared, incubated with 25 mM ethanol for 24, 48, and 72 h, harvested, and evaluated for ethanol metabolism, triglyceride production, oxidative stress, and cytokine expression. Ethanol metabolism and acetaldehyde production decreased in PCLS from obese rats compared with age-matched controls (AMC). Increased triglyceride and smooth muscle actin production was observed in PCLS from obese rats compared with AMC, which further increased following ethanol incubation. Lipid peroxidation, measured by thiobarbituric acid reactive substances assay, increased in response to ethanol, whereas GSH and heme oxygenase I levels were decreased. TNF-α and IL-6 levels were increased in the PCLS from obese rats and increased further with ethanol incubation. Diet-induced fatty liver increases the susceptibility of the liver to toxins such as ethanol, possibly by the increased oxidative stress and cytokine production. These findings support the concept that the development of fatty liver sensitizes the liver to the effects of ethanol and leads to the start of liver failure, necrosis, and eventually cirrhosis.
Assuntos
Etanol/farmacologia , Ácidos Graxos/biossíntese , Fígado/efeitos dos fármacos , Obesidade/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Animais , Dieta Hiperlipídica , Etanol/metabolismo , Fígado Gorduroso/metabolismo , Interleucina-6/metabolismo , Peroxidação de Lipídeos/efeitos dos fármacos , Peroxidação de Lipídeos/fisiologia , Fígado/metabolismo , Masculino , Modelos Animais , Ratos , Ratos Sprague-Dawley , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Biomarkers such as interleukin-6 (IL-6), soluble interleukin-6 receptor (sIL-6R), and high sensitive C-reactive protein (hsCRP) have been reported to be elevated in acute myocardial infarction (AMI). The aim of this study is to determine the relationship between these markers during AMI, as well as their relationship to clinical parameters in an effort to discern their predictive potential in cardiac events. Serum was collected from 73 patients with; AMI, stable coronary artery disease (CAD), and controls during cardiac catheterization. Biomarker levels were determined and correlated with clinical data. IL-6 (11.75pg/ml, P<0.05) and sIL-6R (41,340pg/ml, P=0.05) were elevated in AMI compared with CAD and controls. At presentation, hsCRP was elevated in AMI patients (4.69mg/L) compared to controls (2.69mg/L, P<0.05); however, there was a significant decrease in hsCRP between AMI (4.69mg/L) and CAD patients (7.4mg/L, P<0.05). After 24h post-AMI hsCRP levels were increased compared to stable CAD (60.46mg/L, P<0.05) and were preceded by increased IL-6 at presentation. Soluble Gp130 (sGp130) showed no significant change between AMI, CAD, and control patients. However, sGp130 positively correlated with peak troponin in AMI (R=0.587, P<0.01), and negatively correlated with previous AMI (R=-0.382, P<0.05). Circulating monocyte mRNA expression isolated from selected AMI patients showed an increase in IL-6 mRNA (5.28-fold, P<0.01) and a decrease in both IL-6R (0.374-fold, P<0.01) and sGp130 mRNA (0.38-fold, P<0.01) as compared to CAD and controls. Results demonstrate that IL-6 and sIL-6R are associated with AMI and cardiac injury. These data support the hypothesis that trans-IL-6 receptor binding may alter intracellular signaling, and blocking of IL-6 receptor binding may be pathogenic in AMI. These data may be predictive of mechanism(s) by which plaques become unstable and rupture.
Assuntos
Doença da Artéria Coronariana/sangue , Doença da Artéria Coronariana/genética , Interleucina-6/sangue , Infarto do Miocárdio/sangue , Infarto do Miocárdio/genética , Receptores de Interleucina-6/sangue , Proteína C-Reativa/metabolismo , Estudos de Casos e Controles , Receptor gp130 de Citocina/sangue , Demografia , Ensaio de Imunoadsorção Enzimática , Feminino , Regulação da Expressão Gênica , Humanos , Interleucina-6/genética , Masculino , Pessoa de Meia-Idade , Monócitos/metabolismo , Receptores de Interleucina-6/genética , Análise de RegressãoRESUMO
An adolescent male trauma patient developed new asymptomatic ST segment elevations that mimicked a myocardial infarction on infero-lateral telemetry leads on hospital day #8, following burn excision and skin grafting. This was confirmed on 12 lead electrocardiogram. Laboratory test results indicated normal potassium. Troponins ×3 were negative. X-rays indicated marked gaseous gastric distention. A nasogastric tube was placed with evacuation of 400 mL of fluid and resolution of gastric distention. After gastric decompression, the ST segment elevations resolved. This case illustrates the need to consider acute gastric distention in the differential of acute ST segment elevation.
RESUMO
Cardiovascular disease is a major cause of morbidity and mortality in the world and better prevention and treatment strategies are needed. Studies from this laboratory have shown that perfluorocarbon exposed sonicated dextrose albumin (PESDA) microbubbles bind to inflamed vasculature through interactions with scavenger receptors (SR). This current study details the use of PESDA as a tool for accessing and quantifying the extent of vascular inflammation. Angioplastied rat aortas were evaluated with low mechanical index microbubble imaging techniques contrast pulse sequencing (CPS); Siemens Acuson Sequoia 15L8, 7-15 MHz ultrasound probe with a mechanical index of 0.09 to detect microbubble binding. Real-time polymerase chain reaction (RT-PCR) analysis of angioplastied aortas demonstrated a significantly (p < 0.01) increased expression of both SRs and Interleukin 6 (IL-6). Vessel wall enhancement was quantified using densitometry of CPS ultrasound images and correlated with the upregulated expression of scavenger receptors, Toll-like receptors and IL-6. This study demonstrates that PESDA, in conjunction with CPS ultrasound, is an effective imaging technique to better detect early vascular inflammation and potential cardiovascular disease.
Assuntos
Angioplastia com Balão/efeitos adversos , Aorta/diagnóstico por imagem , Aorta/lesões , Aterosclerose/diagnóstico por imagem , Albuminas/farmacologia , Animais , Aterosclerose/metabolismo , Biomarcadores/metabolismo , Meios de Contraste/farmacologia , Modelos Animais de Doenças , Fluorocarbonos/farmacologia , Inflamação/diagnóstico por imagem , Inflamação/metabolismo , Interleucina-6/metabolismo , Masculino , Microbolhas , Ratos , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase em Tempo Real , Receptores Depuradores/metabolismo , Receptores Toll-Like/metabolismo , UltrassonografiaRESUMO
BACKGROUND: Coronary artery disease is the single leading cause of death in the United States. Commonly it is treated with coronary bypass grafting using the saphenous vein (SV) or internal mammary artery (IMA) as a conduit. Unfortunately, the SV has much lower patency rates compared with the IMA. Several hypotheses exist as to why occlusion occurs more commonly in SV grafts than in IMA grafts. However detailed studies in this area have been limited. This study investigates the effects of pressure distention on inflammation in SV conduit used in coronary artery bypass grafting (CABG). METHODS: Saphenous vein distention pressure was measured intraoperatively during 48 CABG procedures. A segment of SV was excised from the conduit before distention. Because the vein was used for coronary artery grafting, sequential pieces were archived for evaluation. Real-time polymerase chain reaction (RT-PCR) and immunohistochemical analyses were performed to investigate a change in the expression of biomarkers. RESULTS: Upregulation of various biomarkers occurred. These biomarkers included scavenger receptors A and B (SR-A, SR-B), toll-like receptors 2 and 4 (TLR2, TLR4), platelet endothelial cell adhesion molecule (PECAM), vascular cell adhesion molecule (VCAM), and intercellular cell adhesion molecule (ICAM) in segments of SV that were subjected to distention. Immunohistochemical results mirrored RT-PCR findings. A significant correlation was observed between biomarkers and pressure values. CONCLUSIONS: These studies demonstrate that markers of inflammation are upregulated in response to SV distention. The data suggest that the pressure used in graft preparation procedures should be regulated to avoid inflammation and its potential to induce graft failure.
Assuntos
Moléculas de Adesão Celular/análise , Ponte de Artéria Coronária/métodos , Oclusão de Enxerto Vascular/etiologia , Receptores Depuradores/análise , Veia Safena/transplante , Coleta de Tecidos e Órgãos/efeitos adversos , Receptores Toll-Like/análise , Idoso , Biomarcadores , Moléculas de Adesão Celular/biossíntese , Moléculas de Adesão Celular/genética , Doença das Coronárias/cirurgia , Feminino , Oclusão de Enxerto Vascular/metabolismo , Humanos , Hiperplasia , Masculino , Pessoa de Meia-Idade , Flebite/etiologia , Flebite/metabolismo , Pressão/efeitos adversos , RNA Mensageiro/análise , Reação em Cadeia da Polimerase em Tempo Real , Receptores Depuradores/biossíntese , Receptores Depuradores/genética , Veia Safena/metabolismo , Veia Safena/patologia , Coleta de Tecidos e Órgãos/métodos , Receptores Toll-Like/biossíntese , Receptores Toll-Like/genética , Regulação para Cima , Grau de Desobstrução VascularRESUMO
OBJECTIVE: To determine whether serum complement C3 mediates adherence of albumin-encapsulated microbubbles to vascular endothelium in the development of atherosclerotic plaques. METHODS: Adherence of microbubbles to aortic endothelium was examined with scanning electron microscopy following intravenous injection of 20% intralipid in wild-type mice, genetic complement-deficient mice (C3-/-), and in pharmacologic C3-depleted wild-type mice. In a second experimental model, atherosclerostic plaque was induced in apolipoprotein E-deficient mice (apoE-/-), and adherence of microbubbles to atherosclerotic plaques was evaluated using fluorescent microscopy of fluorescein isothiocynate-conjugated microbubbles. Finally, imaging of aortas was performed in eight rats (four JCR:LA-cp atherosclerosis-prone rats on high cholesterol diets; four controls) following intravenous albumin microbubble injections (PESDA) to determine whether microbubble adherence to the endothelium could be detected with low mechanical index pulse sequence schemes. RESULTS: Scanning electron microscopy confirmed the adherence of microbubbles to the endothelial cells of the aorta in wild-type mice following induction of hypertriglyceridemia but not in C3-depleted mice. Microbubble adherence to the endothelial surface of atherosclerotic plaque was confirmed in all apoE-/- mice (median 172 microbubbles/field; compared to a median of 3 microbubbles/field in cobra venom factor-treated apoE-/- mice; p < 0.001). Low mechanical index ultrasound imaging detected microbubble adherence in all JCR atherosclerosis prone rats even in the absence of vasomotor or phenotypical evidence of endothelial dysfunction. The numbers of adherent microbubbles correlated with serum triglyceride levels, and were seen in conjunction with increased endothelial nitric oxide synthase activity. CONCLUSIONS: Complement C3 binds to albumin-encapsulated microbubbles and mediates microbubble adherence to vascular endothelium both early and late in the atherosclerotic process.
Assuntos
Albuminas/metabolismo , Aterosclerose/metabolismo , Complemento C3/metabolismo , Endotélio Vascular/metabolismo , Hipertrigliceridemia/metabolismo , Animais , Aorta , Apolipoproteínas E/deficiência , Aterosclerose/patologia , Complemento C3/genética , Venenos Elapídicos/farmacologia , Endotélio Vascular/ultraestrutura , Feminino , Fluoresceína-5-Isotiocianato , Corantes Fluorescentes , Hipertrigliceridemia/patologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Microbolhas , Microscopia Confocal , Microscopia Eletrônica de Varredura , Ratos , Aderências Teciduais , Triglicerídeos/sangueRESUMO
Translation initiation of coxsackievirus B3 (CVB3) RNA is directed by an internal ribosome entry site (IRES) within the 5' untranslated region. However, the details of ribosome-template recognition and subsequent translation initiation are still poorly understood. In this study, we have provided evidence to support the hypothesis that 40S ribosomal subunits bind to CVB3 RNA via basepairing with 18S rRNA in a manner analogous to that of the Shine-Dalgarno (S-D) sequence in prokaryotic systems. We also identified a new site within both the 18S rRNA and the polpyrimidine-tract sequence of the IRES that allows them to form stronger sequence complementation. All these data were obtained from in vitro translation experiments using mutant RNAs containing either an antisense IRES core sequence at the original position or site-directed mutations or deletions in the polypyrimidine tract of the IRES. The mutations significantly reduced translation efficiency but did not abolish protein synthesis, suggesting that the S-D-like sequence is essential, but not sufficient for ribosome binding. To determine how ribosomes reach the initiation codon after internal entry, we created additional mutants: when the authentic initiation codon at nucleotide (nt) 742 was mutated, a 180-nt downstream in-frame AUG codon at nt 922 is able to produce a truncated smaller protein. When this mutation was introduced into the full-length cDNA of CVB3, the derived viruses were still infectious. However, their infectivity was much weaker than that of the wild-type CVB3. In addition, when a stable stem-loop was inserted upstream of the initiation codon in the bicistronic RNA, translation was strongly inhibited. These data suggest that ribosomes reach the initiation codon from the IRES likely by scanning along the viral RNA.