RESUMEN
Mycobacterium chimaera causes pulmonary disease, but little is known of gradations in isolate virulence. Previously, 17 M. chimaera isolates were screened for survival in THP1 macrophages. "M. chimaera 1" was categorized as "more virulent" because it showed the greatest survival in macrophages, whereas "M. chimaera 2" was categorized as "less virulent" with reduced survival. Herein, we infected C3HeB/FeJ mice to compare the in vivo immune responses to M. chimaera 1 and 2. Unlike macrophages, significantly lower M. chimaera 1 counts were recovered from mouse lung tissue and BAL cells with less lung histopathologic changes compared to M. chimaera 2. Compared to M. chimaera 2, significantly more IL-1ß, IL-6, and TNFα was produced early after M. chimaera 1 infection. LC-MS metabolomics analyses of BAL fluid revealed divergence in sphingolipid, phospholipid metabolism between M. chimaera 1 versus M. chimaera 2 mice. From pan-GWAS analyses, virulence and organizing DNA/molecular structure genes were associated with more virulent M. chimaera isolates. Vigorous lung-specific immune responses to M. chimaera 1 may influence effective bacterial control, but for a different isolate M. chimaera 2, subvert immune control. Continued studies of the gradations in virulence among the same NTM species will advance our understanding of NTM pathogenesis.
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Infecciones por Mycobacterium , Mycobacterium , Animales , Ratones , Modelos Animales de Enfermedad , Infecciones por Mycobacterium/microbiología , Mycobacterium/genética , PulmónRESUMEN
The link between military deployment to Southwest Asia and Afghanistan, and the risk for lung disease, including bronchiolitis, is increasingly well-recognized. However, histopathologic features that distinguish deployment-related lung diseases from other diseases affecting the small airways and airspaces are uncertain. A computer-based scoring system was developed to characterize surgical lung biopsy findings in 65 soldiers with persistent respiratory symptoms following military deployment ("deployers"). Deployer lung biopsies were compared to those from 8 patients with chronic hypersensitivity pneumonitis (cHP), 10 with smoking-related respiratory bronchiolitis, 11 with autoimmune or post-transplant obliterative bronchiolitis, and 10 normal donor lungs. Upper, middle, and lower lobe-specific findings in deployer samples were analyzed to inform optimum biopsy location choice for future patients. Surgical lung biopsies from symptomatic deployed military service members were distinguished by a combination of small airways abnormalities including smooth muscle hypertrophy (SMH), peribronchiolar metaplasia (PBM), and lymphocytic inflammation, often with constrictive/obliterative (C/O) and/or respiratory bronchiolitis (43.1%), granulomatous inflammation (38.5%), and moderate/severe emphysema (46.2%, mainly in nonsmokers). Lymphocytic pleural inflammation was common (89.2%), and vascular abnormalities occurred in nearly one-third. Histopathologic features in deployers were most strongly overlapping with cases of cHP, both showing granulomatous inflammation, PBM, and emphysema. SMH along with C/O and respiratory bronchiolitis were common in deployers but not in cHP cases. In deployers, there were significantly higher odds of small airways injury in the lower lobe compared with upper lobe samples.
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Bronquiolitis , Enfisema , Enfermedades Pulmonares , Personal Militar , Bronquiolitis/patología , Enfisema/patología , Humanos , Inflamación/patología , Pulmón/patología , Enfermedades Pulmonares/patologíaRESUMEN
BACKGROUND: Pulmonary vascular remodeling in pulmonary hypertension (PH) is characterized by increased vascular smooth muscle cell (SMC) and adventitial fibroblast proliferation, small vessel occlusion, and inflammatory cell accumulation. The underlying molecular mechanisms driving progression remain poorly defined. We have focused on loss of the phosphatase PTEN in SMCs as a major driver of pathological vascular remodeling. Our goal was to define the role of PTEN in human PH and in hypoxia-induced PH using a mouse model with inducible deletion of PTEN in SMCs. METHODS AND RESULTS: Staining of human biopsies demonstrated enhanced inactive PTEN selectively in the media from hypertensive patients compared to controls. Mice with induced deletion of PTEN in SMCs were exposed to normoxia or hypoxia for up to 4 weeks. Under normoxia, SMC PTEN depletion was sufficient to induce features of PH similar to those observed in wild-type mice exposed to chronic hypoxia. Under hypoxia, PTEN depletion promoted an irreversible progression of PH characterized by increased pressure, extensive pulmonary vascular remodeling, formation of complex vascular lesions, and increased macrophage accumulation associated with synergistic increases in proinflammatory cytokines and proliferation of both SMCs and nonSMCs. CONCLUSIONS: Chronic inactivation of PTEN selectively in SMC represents a critical mediator of PH progression, leading to cell autonomous events and increased production of factors correlated to proliferation and recruitment of adventitial and inflammatory cells, resulting in irreversible progression of the disease.
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Hipertensión Pulmonar/etiología , Hipoxia/complicaciones , Fosfohidrolasa PTEN/fisiología , Animales , Progresión de la Enfermedad , Masculino , Ratones , Miocitos del Músculo Liso , Ratas , Índice de Severidad de la EnfermedadRESUMEN
RATIONALE: Severe pulmonary arterial hypertension (SPH) is a frequently lethal condition characterized by pulmonary vascular remodeling and right heart strain or failure. SPH is also often associated with autoimmune and collagen vascular disorders. OBJECTIVES: To study the effects of T cells on the development of experimental SPH. METHODS: Athymic nude rats lacking T cells were treated with a single subcutaneous injection of vascular endothelial growth factor (VEGF) receptor blocker SU5416 (20 mg/kg) to induce pulmonary vascular endothelial cell apoptosis. Immunohistochemical analysis and IL-4 levels of the lung tissue were performed. Cell death and proliferation were assessed by Western blot and immunohistochemistry. MEASUREMENTS AND MAIN RESULTS: In contrast to SU5416-treated euthymic rats that develop SPH only in combination with chronic hypoxia, athymic nude rats developed SPH and vascular remodeling (similar to clinical SPH) at normoxic conditions as demonstrated by measurements of pulmonary artery pressure and right ventricle hypertrophy. Pulmonary arterioles became occluded with proliferating endothelial cells and were surrounded by mast cells, B cells, and macrophages. IL-4, proliferating cell nuclear antigen, and collagen type I levels were markedly increased in SU5416-treated athymic rat lungs. Antibody deposition was noted along the vascular endothelium in rats with SPH. Finally, protection from SPH was conferred by immune challenge with spleen cells from euthymic nude rats. CONCLUSIONS: These studies demonstrate the importance of a complete, intact immune system in protecting against pulmonary angioproliferation in this new model of SPH as well as the importance of intact VEGF receptor signaling for lung endothelial cell homeostasis.
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Hipertensión Pulmonar/inmunología , Linfocitos T/inmunología , Traslado Adoptivo , Inhibidores de la Angiogénesis , Animales , Células Dendríticas/inmunología , Modelos Animales de Enfermedad , Células Endoteliales , Homeostasis/inmunología , Inmunohistoquímica , Indoles/farmacología , Inyecciones Subcutáneas , Masculino , Pirroles/farmacología , Ratas , Ratas Desnudas , Receptores de Factores de Crecimiento Endotelial Vascular/antagonistas & inhibidores , Receptores de Factores de Crecimiento Endotelial Vascular/inmunologíaRESUMEN
RATIONALE: Impaired endothelial cell-dependent vasodilation, inflammation, apoptosis, and proliferation are manifestations of endothelial dysfunction in chronic obstructive pulmonary disease (COPD). Prostacyclin (PGI(2)) is a major product of the cyclooxygenase pathway with potent vasodilatory and antimitogenic properties and may be relevant to endothelial dysfunction in COPD. OBJECTIVES: To determine if PGI(2) expression is altered in smoking-related lung disease and if it may be protective in COPD-associated endothelial dysfunction. METHODS: We evaluated, by immunohistochemistry, Western blotting, and polymerase chain reaction, human emphysema tissue compared with normal tissue for expression of prostacyclin synthase (PGI(2)S). We examined the effects of cigarette smoke extract (CSE) and aldehyde components on eicosanoid expression in primary human pulmonary microvascular endothelial cells. Finally, we used a murine model of lung-specific PGI(2)S overexpression and in vitro studies to determine if PGI(2) expression has protective effects on cigarette smoke-induced endothelial apoptosis. MEASUREMENTS AND MAIN RESULTS: Human emphysema lung tissue exhibited lower PGI(2)S expression within the pulmonary endothelium than in normal lung. In vitro studies demonstrated that CSE, and in particular the alpha,beta unsaturated aldehyde acrolein, suppressed PGI(2)S gene expression, whereas CSE significantly induced the upstream mediators COX-2 and cytosolic phospholipase A2 in human pulmonary microvascular endothelial cells. Mice with lung-specific PGI(2)S overexpression exhibited less endothelial apoptosis after chronic smoke exposure. In vitro, iloprost exhibited protective effects on CSE-induced apoptosis. CONCLUSIONS: PGI(2) has protective effects in the pulmonary vasculature after acute and chronic cigarette smoke exposure. An imbalance in eicosanoid expression may be important to COPD-associated endothelial dysfunction.
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Acroleína/farmacología , Endotelio Vascular/efectos de los fármacos , Epoprostenol/fisiología , Enfisema Pulmonar/metabolismo , Enfisema Pulmonar/prevención & control , Fumar/efectos adversos , Animales , Apoptosis/efectos de los fármacos , Estudios de Casos y Controles , Técnicas de Cultivo de Célula , Células Endoteliales/efectos de los fármacos , Células Endoteliales/fisiología , Endotelio Vascular/patología , Humanos , Pulmón/irrigación sanguínea , Pulmón/efectos de los fármacos , Pulmón/patología , Ratones , Ratones Transgénicos , Enfisema Pulmonar/etiologíaRESUMEN
RATIONALE: HIV-infected patients with pulmonary arterial hypertension have histologic manifestations that are indistinguishable from those found in patients with idiopathic pulmonary arterial hypertension. In addition, the role of pleiotropic viral proteins in the development of plexiform lesions in HIV-related pulmonary hypertension (HRPH) has not been explored. Simian immunodeficiency virus (SIV) infection of macaques has been found to closely recapitulate many of the characteristic features of HIV infection, and thus hallmarks of pulmonary arterial hypertension should also be found in this nonhuman primate model of HIV. OBJECTIVES: To determine whether pulmonary arterial lesions were present in archived SIV-infected macaque lung tissues from Johns Hopkins University and two National Primate Research Centers. METHODS: Archived macaque and human lung sections were examined via immunohistochemistry for evidence of complex vascular lesions. RESULTS: Complex plexiform-like lesions characterized by lumenal obliteration, intimal disruption, medial hypertrophy, thrombosis, and recanalized lumena were found exclusively in animals infected with SHIV-nef (a chimeric viral construct containing the HIV nef gene in an SIV backbone), but not in animals infected with SIV. The mass of cells in the lesions were factor VIII positive, and contained cells positive for muscle-specific and smooth muscle actins. Lung mononuclear cells were positive for HIV Nef, suggesting viral replication. Endothelial cells in both the SHIV-nef macaques and patients with HRPH, but not in patients with idiopathic pulmonary arterial hypertension, were also Nef positive. CONCLUSIONS: The discovery of complex vascular lesions in SHIV-nef- but not SIV-infected animals, and the presence of Nef in the vascular cells of patients with HRPH, suggest that Nef plays a key role in the development of severe pulmonary arterial disease.
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Productos del Gen nef/fisiología , VIH-1/genética , Hipertensión Pulmonar/genética , Síndrome de Inmunodeficiencia Adquirida del Simio/genética , Proteínas Reguladoras y Accesorias Virales/genética , Animales , Modelos Animales de Enfermedad , Células Epiteliales/metabolismo , Técnica del Anticuerpo Fluorescente , VIH-1/fisiología , Humanos , Inmunohistoquímica , Macaca , Virus de la Inmunodeficiencia de los Simios/genética , Virus de la Inmunodeficiencia de los Simios/patogenicidad , Replicación Viral , Productos del Gen nef del Virus de la Inmunodeficiencia HumanaRESUMEN
Pigment epithelium-derived factor (PEDF) is a 50-kD protein with angiostatic and neurotrophic activities that regulates vascular development within the eye. PEDF expression was increased in the lungs of patients with idiopathic pulmonary fibrosis (IPF) based on microarray analyses. Angiogenesis has been implicated in the pathogenesis of fibrotic lung diseases, we therefore hypothesized that regional abnormalities in vascularization occur in IPF as a result of an imbalance between PEDF and vascular endothelial growth factor. We demonstrated that vascular density is regionally decreased in IPF within the fibroblastic foci, and that within these areas PEDF was increased, whereas vascular endothelial growth factor was decreased. PEDF colocalized with the fibrogenic cytokine, transforming growth factor (TGF)-beta 1, particularly within the fibrotic interstitium and the fibroblastic focus, and prominently within the epithelium directly overlying the fibroblastic focus. This suggested that TGF-beta 1 might regulate PEDF expression. Using 3T3-L1 fibroblasts and human lung fibroblasts, we showed that PEDF was indeed a TGF-beta 1 target gene. Collectively, our findings implicate PEDF as a regulator of pulmonary angiogenesis and an important mediator in IPF.
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Proteínas del Ojo , Neovascularización Patológica/metabolismo , Neovascularización Patológica/patología , Factores de Crecimiento Nervioso , Proteínas/metabolismo , Fibrosis Pulmonar/metabolismo , Fibrosis Pulmonar/patología , Serpinas/metabolismo , Biopsia , Líquido del Lavado Bronquioalveolar/química , Fibroblastos/efectos de los fármacos , Fibroblastos/metabolismo , Humanos , Pulmón/metabolismo , Pulmón/patología , Enfermedades Pulmonares Intersticiales/metabolismo , Enfermedades Pulmonares Intersticiales/fisiopatología , Proteínas/efectos de los fármacos , Serpinas/efectos de los fármacos , Factor de Crecimiento Transformador beta/metabolismo , Factor de Crecimiento Transformador beta/farmacología , Factor de Crecimiento Transformador beta1 , Factor A de Crecimiento Endotelial Vascular/metabolismoRESUMEN
Competency models, a combination of identified skills, knowledge and attributes required for successful role performance, lend a roadmap for success to leaders of today and the new leaders of tomorrow. In healthcare, where the environment is ever-changing, and the pool of qualified leaders is limited and shrinking, competency models help to identify required areas of strength for success in performance. The Peterborough Regional Health Centre (PRHC) underwent a leadership reorganization in 2001, structuring all leaders into positions based on a corporate competency model. Leaders completed self-assessment portfolios and were shifted into roles emphasizing job-person fit, competencies, personal preference and organizational principles and accountabilities. The literature suggests that evaluations are rarely performed after corporate reorganizations, and when they are, they rarely measure indicators that are directly related to outcomes. In response to this, PRHC has structured an evaluation of the reorganization in two phases. The first phase is focused on process changes that are a direct result of the reorganization. The second phase, ongoing over 24 months, is focused on outcomes that are resulting from the reorganization.