RESUMEN
Acute lung injury (ALI) and acute respiratory distress syndrome (ARDS) are severe complications that can occur in infections caused by any Plasmodium species. Due to the high lethality rate and the lack of specific treatment for ALI/ARDS, studies aimed at understanding and searching for treatment strategies for such complications have been fundamental. Here, we investigated the protective role of dietary supplementation with DHA-rich fish oil against lung damage induced by Plasmodium berghei ANKA in a murine model. Our results demonstrated that alveolar vascular damage, lung edema, and histopathological alterations were significantly reduced in mice that received dietary supplementation compared to those that did not receive the supplementation. Furthermore, a significant reduction in the number of CD8+ T lymphocytes, in addition to reduced infiltration of inflammatory cells in the bronchoalveolar lavage fluid was also observed. High levels of IL-10, but not of TNF-α and IFN-γ, were also observed in infected mice that received the supplementation, along with a reduction in local oxidative stress. Together, the data suggest that dietary supplementation with DHA-rich fish oil in malarial endemic areas may help reduce lung damage resulting from the infection, thus preventing worsening of the condition.
Asunto(s)
Suplementos Dietéticos , Modelos Animales de Enfermedad , Ácidos Docosahexaenoicos , Malaria , Plasmodium berghei , Animales , Plasmodium berghei/efectos de los fármacos , Ratones , Ácidos Docosahexaenoicos/farmacología , Ácidos Docosahexaenoicos/administración & dosificación , Pulmón/patología , Pulmón/efectos de los fármacos , Pulmón/parasitología , Líquido del Lavado Bronquioalveolar/química , Ácidos Grasos Omega-3/farmacología , Ácidos Grasos Omega-3/administración & dosificación , Estrés Oxidativo/efectos de los fármacos , Lesión Pulmonar Aguda/prevención & control , Lesión Pulmonar Aguda/tratamiento farmacológico , Linfocitos T CD8-positivos/inmunología , Interleucina-10 , Aceites de Pescado/farmacología , Aceites de Pescado/administración & dosificaciónRESUMEN
Atmospheric pollution can be defined as a set of changes that occur in the composition of the air, making it unsuitable and/or harmful and thereby generating adverse effects on human health. The regular practice of physical exercise (PE) is associated with the preservation and/or improvement of health; however, it can be influenced by neuroimmunoendocrine mechanisms and external factors such as air pollution, highlighting the need for studies involving the practice of PE in polluted environments. Herein, 24 male C57BL/6 mice were evaluated, distributed into four groups (exposed to a high concentration of pollutants/sedentary, exposed to a high concentration of pollutants/exercised, exposed to ambient air/sedentary, and exposed to ambient air/exercised). The exposure to pollutants occurred in the environmental particle concentrator (CPA) and the physical training was performed on a treadmill specially designed for use within the CPA. Pro- and anti-inflammatory markers in blood and bronchoalveolar lavage (BALF), BALF cellularity, and lung tissue were evaluated. Although the active group exposed to a high concentration of pollution showed a greater inflammatory response, both the correlation analysis and the ratio between pro- and anti-inflammatory cytokines demonstrated that the exercised group presented greater anti-inflammatory activity, suggesting a protective/adaptative effect of exercise when carried out in a polluted environment.
Asunto(s)
Contaminantes Atmosféricos , Líquido del Lavado Bronquioalveolar , Citocinas , Ratones Endogámicos C57BL , Condicionamiento Físico Animal , Animales , Masculino , Ratones , Líquido del Lavado Bronquioalveolar/química , Líquido del Lavado Bronquioalveolar/inmunología , Citocinas/metabolismo , Contaminantes Atmosféricos/toxicidad , Contaminantes Atmosféricos/análisis , Inflamación/inducido químicamente , Pulmón/efectos de los fármacos , Contaminación del Aire/análisis , Contaminación del Aire/efectos adversos , Material Particulado/toxicidad , Material Particulado/análisisRESUMEN
Aspergillus species can colonize and infect immunocompetent and immunocompromised hosts. Conventional fungal identification depends on microscopic analysis and microorganism medium growth. Other diagnostic methods, non-growth dependent, to invasive fungal infections, are the biomarkers that detect circulating polysaccharides, for example, 1-3-ß-d-Glucan and galactomannan. Both are polysaccharides present on the external layer of fungi cell wall and can be detected in clinical samples during the growth of the fungus in the patient. This study aimed to compare the galactomannan detection of Lateral Flow Assay and Enzyme Immunoassay methods in Bronchoalveolar Lavage Fluid. The galactomannan antigen in Bronchoalveolar Lavage Fluid was measured using Enzyme Immunoassay according to the manufacturer's instructions (PLATELIA ASPERGILLUS™ BioRad) and, using a Lateral Flow Assay according to the manufacturer's instructions (Galactomannan LFA IMMY©). The 71 samples were Bronchoalveolar Lavage Fluid of patients hospitalized at Unicamp Clinical Hospital between 2019 and 2021; of these samples 12/71 (16.9 %) resulted in positive Galactomannan-Lateral Flow Assay. In contrast, Galactomannan-Enzyme Immunoassay resulted as positive in 9/71 (12.6 %) samples, a difference that showed not significant statistically (p-value = 0.36) Comparing both assays' results identified 8 divergences between them, about 11 % of the total sample. The Sensitivity (73.3 %), Specificity (92.35 %), Positive Predictive Value (62.85 %) and Negative Predictive Value (95.15 %) of Lateral Flow Assay were calculated using the Galactomannan Enzyme Immunoassay as standard. The Lateral Flow Assay demonstrated good results when compared with the Enzyme Immunoassay.
Asunto(s)
Aspergillus , Líquido del Lavado Bronquioalveolar , Galactosa , Técnicas para Inmunoenzimas , Mananos , Sensibilidad y Especificidad , Mananos/análisis , Galactosa/análogos & derivados , Humanos , Líquido del Lavado Bronquioalveolar/microbiología , Líquido del Lavado Bronquioalveolar/química , Aspergillus/inmunología , Aspergillus/aislamiento & purificación , Técnicas para Inmunoenzimas/métodos , Aspergilosis/diagnóstico , Aspergilosis/microbiología , Biomarcadores/análisis , Antígenos Fúngicos/análisis , Reproducibilidad de los ResultadosRESUMEN
Prolonged exposure to iron powder and other mineral dusts can threaten the health of individuals, especially those with COPD. The goal of this study was to determine how environmental exposure to metal dust from two different mining centers in Brazil affects lung mechanics, inflammation, remodeling and oxidative stress responses in healthy and elastase-exposed mice. This study divided 72 male C57Bl/6 mice into two groups, the summer group and the winter group. These groups were further divided into six groups: control, nonexposed (SAL); nonexposed, given elastase (ELA); exposed to metal powder at a mining company (SAL-L1 and ELA-L1); and exposed to a location three miles away from the mining company (SAL-L2 and ELA-L2) for four weeks. On the 29th day of the protocol, the researchers assessed lung mechanics, bronchoalveolar lavage fluid (BALF), inflammation, remodeling, oxidative stress, macrophage iron and alveolar wall alterations (mean linear intercept-Lm). The Lm was increased in the ELA, ELA-L1 and ELA-L2 groups compared to the SAL group (p < 0.05). There was an increase in the total number of cells and macrophages in the ELA-L1 and ELA-L2 groups compared to the other groups (p < 0.05). Compared to the ELA and SAL groups, the exposed groups (ELA-L1, ELA-L2, SAL-L1, and SAL-L2) exhibited increased expression of IL-1ß, IL-6, IL-10, IL-17, TNF-α, neutrophil elastase, TIMP-1, MMP-9, MMP-12, TGF-ß, collagen fibers, MUC5AC, iNOS, Gp91phox, NFkB and iron positive macrophages (p < 0.05). Although we did not find differences in lung mechanics across all groups, there were low to moderate correlations between inflammation remodeling, oxidative stress and NFkB with elastance, resistance of lung tissue and iron positive macrophages (p < 0.05). Environmental exposure to iron, confirmed by evaluation of iron in alveolar macrophages and in air, exacerbated inflammation, initiated remodeling, and induced oxidative stress responses in exposed mice with and without emphysema. Activation of the iNOS, Gp91phox and NFkB pathways play a role in these changes.
Asunto(s)
Exposición a Riesgos Ambientales , Hierro , Elastasa Pancreática , Animales , Masculino , Ratones , Líquido del Lavado Bronquioalveolar/química , Exposición a Riesgos Ambientales/efectos adversos , Inflamación/metabolismo , Inflamación/inducido químicamente , Hierro/toxicidad , Pulmón/efectos de los fármacos , Pulmón/metabolismo , Pulmón/patología , Ratones Endogámicos C57BL , Estrés Oxidativo/efectos de los fármacos , Elastasa Pancreática/metabolismo , Elastasa Pancreática/farmacología , Polvos/toxicidadRESUMEN
Neutrophilic asthma is generally defined by poorly controlled symptoms and high levels of neutrophils in the lungs. Short-chain fatty acids (SCFAs) are proposed as nonpharmacological therapy for allergic asthma, but their impact on the neutrophilic asthma lacks evidence. SCFAs regulate immune cell responses and impact the inflammasome NLRP3, a potential pharmacological target for neutrophilic asthma. Here, we explored the capacity of SCFAs to mitigate murine-induced neutrophilic asthma and the contribution of NLRP3 to this asthma. The objective of this study is to analyze whether SCFAs can attenuate lung inflammation and tissue remodeling in murine neutrophilic asthma and NLRP3 contribution to this endotype. Wild-type (WT) C57BL6 mice orotracheally received 10 µg of HDM (house dust mite) in 80 µL of saline on days 0, 6-10. To explore SCFAs, each HDM group received 200 mM acetate, propionate, or butyrate. To explore NLRP3, Nlrp3 KO mice received the same protocol of HDM. On the 14th day, after euthanasia, bronchoalveolar lavage fluid (BALF) and lungs were collected to evaluate cellularity, inflammatory cytokines, and tissue remodeling. HDM group had increased BALF neutrophil influx, TNF-α, IFN-γ, IL-17A, collagen deposition, and mucus secretion compared to control. SCFAs distinctively attenuate lung inflammation. Only features of tissue remodeling were Nlrp3-dependent such as collagen deposition, mucus secretion, active TGF-ß cytokine, and IMs CD206+. SCFAs greatly decreased inflammatory cytokines and tissue remodeling. Only tissue remodeling was dependent on NLRP3. It reveals the potential of SCFAs to act as an additional therapy to mitigate neutrophilic asthma and the NLRP3 contribution to asthma.
Asunto(s)
Asma , Ácidos Grasos Volátiles , Ratones Endogámicos C57BL , Proteína con Dominio Pirina 3 de la Familia NLR , Neutrófilos , Neumonía , Animales , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismo , Asma/metabolismo , Asma/inmunología , Asma/tratamiento farmacológico , Ratones , Neutrófilos/inmunología , Neutrófilos/metabolismo , Ácidos Grasos Volátiles/metabolismo , Neumonía/metabolismo , Neumonía/inmunología , Ratones Noqueados , Pyroglyphidae/inmunología , Pulmón/patología , Pulmón/metabolismo , Pulmón/inmunología , Remodelación de las Vías Aéreas (Respiratorias)/efectos de los fármacos , Citocinas/metabolismo , Líquido del Lavado Bronquioalveolar/citología , Líquido del Lavado Bronquioalveolar/químicaRESUMEN
Mechanical ventilation is an essential supportive therapy in the treatment of critical patients, and it aims to maintain adequate gas exchange; however, it can also contribute to inflammation and oxidative stress, thus leading to lung injury. We tested the hypothesis that exogenous surfactant administration will be protective against ventilator-induced lung injury in adult healthy Wistar rats both because of its anti-inflammatory properties as well as its role in preventing alveolar collapse at end-expiration. Thus, the effect of intranasal instillation of a bovine exogenous surfactant was tested in Wistar rats submitted to mechanical ventilation. The animals were divided into four groups: (1) CONTROL; (2) SURFACTANT; (3) Mechanical ventilation (MV); (4) MV with pre-treatment with surfactant (MVSURFACTANT). The MV and MVSURFACTANT were submitted to MV with high tidal volume (12 mL/kg) for 1 h. After the experimental protocol, all animals were euthanized and the arterial blood, bronchoalveolar lavage fluid and lungs were collected for biochemical, immunoenzymatic assay, arterial blood gases, and morphometric analyzes. The Wistar rats that received exogenous surfactant (Survanta®) by intranasal instillation before MV demonstrated reduced levels of leukocytes, inflammatory biomarkers such as CCL2, IL-1, IL-6 and TNF-α. Furthermore, it prevented oxidative damage by reducing lipid peroxidation and protein carbonylation as well as histological pattern changes of pulmonary parenchyma. Our data indicate that exogenous surfactant attenuated lung inflammation and redox imbalance induced by mechanical ventilation in healthy adult rats suggesting a preventive effect on ventilator-induced lung injury.
Asunto(s)
Surfactantes Pulmonares , Lesión Pulmonar Inducida por Ventilación Mecánica , Animales , Líquido del Lavado Bronquioalveolar/química , Bovinos , Humanos , Pulmón , Surfactantes Pulmonares/metabolismo , Surfactantes Pulmonares/farmacología , Ratas , Ratas Wistar , Respiración Artificial , Tensoactivos/farmacología , Tensoactivos/uso terapéutico , Lesión Pulmonar Inducida por Ventilación Mecánica/tratamiento farmacológico , Lesión Pulmonar Inducida por Ventilación Mecánica/prevención & controlRESUMEN
COVID-19 is a disease with a spectrum of clinical responses ranging from moderate to critical. To study and control its effects, a large number of researchers are focused on two substantial aims. On the one hand, the discovery of diverse biomarkers to classify and potentially anticipate the disease severity of patients. These biomarkers could serve as a medical criterion to prioritize attention to those patients with higher prone to severe responses. On the other hand, understanding how the immune system orchestrates its responses in this spectrum of disease severities is a fundamental issue required to design new and optimized therapeutic strategies. In this work, using single-cell RNAseq of bronchoalveolar lavage fluid of nine patients with COVID-19 and three healthy controls, we contribute to both aspects. First, we presented computational supervised machine-learning models with high accuracy in classifying the disease severity (moderate and severe) in patients with COVID-19 starting from single-cell data from bronchoalveolar lavage fluid. Second, we identified regulatory mechanisms from the heterogeneous cell populations in the lungs microenvironment that correlated with different clinical responses. Given the results, patients with moderate COVID-19 symptoms showed an activation/inactivation profile for their analyzed cells leading to a sequential and innocuous immune response. In comparison, severe patients might be promoting cytotoxic and pro-inflammatory responses in a systemic fashion involving epithelial and immune cells without the possibility to develop viral clearance and immune memory. Consequently, we present an in-depth landscape analysis of how transcriptional factors and pathways from these heterogeneous populations can regulate their expression to promote or restrain an effective immune response directly linked to the patients prognosis.
Asunto(s)
Líquido del Lavado Bronquioalveolar/citología , Líquido del Lavado Bronquioalveolar/inmunología , COVID-19/patología , Pulmón/citología , SARS-CoV-2/inmunología , Linfocitos B/inmunología , Biomarcadores , Líquido del Lavado Bronquioalveolar/química , Células Dendríticas/inmunología , Células Epiteliales/citología , Células Epiteliales/virología , Humanos , Células Asesinas Naturales/inmunología , Pulmón/química , Aprendizaje Automático , Macrófagos/inmunología , Monocitos/inmunología , Neutrófilos/inmunología , ARN Viral/genética , Análisis de Secuencia de ARN , Índice de Severidad de la Enfermedad , Análisis de la Célula Individual , Linfocitos T/inmunologíaRESUMEN
BACKGROUND: The knowledge on bovine vaccines against respiratory viruses on bronchoalveolar fluid cells is scarce. OBJECTIVE: To compare the effects of a commercial intranasal (IN) and intramuscular (IM) vaccine against bovine respiratory disease (BRD) complex viruses on bronchoalveolar fluid cells of healthy heifers. METHODS: 21 healthy heifers were assigned to three treatment groups: control (CO, N = 7), intranasally vaccinated (IN) (n = 7), and intramuscularly vaccinated (IM) (n = 7). The IN group received 1 mL of the commercial vaccine in each nostril once containing attenuated BoHV-1, bPIV-3, and BRSV. The IM group was vaccinated with two doses of 2 mL with an interval of 21 days of the commercial vaccine containing attenuated BoHV-1, bPIV-3, and BRSV plus inactivated BVDV. At day 0 (D0), before the first vaccine dose, and at D3, D7, and D21, after the last vaccine dose, airway bronchoscopy was performed to observe local irritation and collect bronchoalveolar lavage fluid (BALF). The bronchoalveolar count, cytological evaluation, bronchoalveolar cell oxidative metabolism, and total bronchoalveolar IgA and IgG were measured. RESULTS: The IN vaccine increased neutrophil cellularity at D7 and D21 and total IgA at D3 in BALF. Total IgA in BALF also increased at D3 and oxidative metabolism of bronchoalveolar cells at D21 lowered compared to the CO group. Following IM vaccination there was no alteration of immunoglobulins or cell oxidative metabolism in BALF. Both vaccines reduced the number of alveolar macrophages. CONCLUSION: Both vaccines induced bronchoalveolar inflammation during the establishment of the vaccine immunity, which was more expressive in the IN protocol.
Asunto(s)
Líquido del Lavado Bronquioalveolar/citología , Enfermedades de los Bovinos/prevención & control , Vacunación/veterinaria , Administración Intranasal/efectos adversos , Administración Intranasal/veterinaria , Animales , Diarrea Mucosa Bovina Viral/prevención & control , Líquido del Lavado Bronquioalveolar/química , Bovinos , Enfermedades de los Bovinos/virología , Virus de la Diarrea Viral Bovina , Femenino , Infecciones por Herpesviridae/prevención & control , Herpesvirus Bovino 1 , Inmunoglobulina A , Inmunoglobulina G , Inyecciones Intramusculares/efectos adversos , Inyecciones Intramusculares/veterinaria , Virus de la Parainfluenza 3 Bovina , Infecciones por Virus Sincitial Respiratorio/prevención & control , Infecciones por Virus Sincitial Respiratorio/veterinaria , Virus Sincitial Respiratorio Bovino , Infecciones por Respirovirus/prevención & control , Infecciones por Respirovirus/veterinaria , Vacunas Atenuadas/administración & dosificación , Vacunas Atenuadas/efectos adversos , Vacunas Virales/administración & dosificaciónRESUMEN
Management of SARS-CoV-2 requires safe decision-making to minimize contamination. Healthcare workers and professionals in confined areas are affected by the risk of the activity and the environment. Isolation of contaminated workers and healthcare professionals requires clinical and diagnostic criteria. On the other hand, interrupting the isolation of healthcare employees and professionals is critical because diagnostic tests do not support clinical decisions. In addition to defining the best test in view of its accuracy, it is necessary to consider aspects such as the stage of the disease or cure, the viral load and the individual's own immunity. Uncertainty about natural and herd immunity to the disease leads to the development of appropriate antivirals, diagnostic tests and vaccines.
Asunto(s)
Prueba de Ácido Nucleico para COVID-19 , Prueba Serológica para COVID-19 , COVID-19/transmisión , Aislamiento de Pacientes/normas , Inmunidad Adaptativa/inmunología , Anticuerpos Antivirales/inmunología , Antígenos Virales/inmunología , Líquido del Lavado Bronquioalveolar/química , Líquido del Lavado Bronquioalveolar/virología , COVID-19/diagnóstico , COVID-19/inmunología , COVID-19/prevención & control , Prueba de COVID-19 , Toma de Decisiones Clínicas , Heces/química , Heces/virología , Personal de Salud , Humanos , Inmunoglobulina G/inmunología , Inmunoglobulina M/inmunología , Nasofaringe/química , Nasofaringe/virología , Aislamiento de Pacientes/métodos , ARN Viral/análisis , SARS-CoV-2 , Esputo/química , Esputo/virología , Carga ViralRESUMEN
Cocoa contains bioactive components, which vary according to genetic and environmental factors. The present study aimed to ascertain the anti-allergic properties of native Peruvian cocoa populations ("Blanco de Piura" or BPC, "Amazonas Peru" or APC, "Criollo de Montaña" or CMC, "Chuncho" or CCC, and an ordinary cocoa or OC). To do so, after an initial in vitro approach, an in vivo study focused on the induction of an anaphylactic response associated with allergic asthma in Brown Norway rats was carried out. Based on their polyphenol content, antioxidant activity and in vitro effects, the APC and CMC were selected to be included in the in vivo study. Cocoa diets were tested in a model of allergic asthma in which anaphylactic response was assessed by changes in body temperature, motor activity and body weight. The concentration of specific immunoglobulin E (IgE), mast cell protease and leukotrienes was also quantified in serum and/or bronchoalveolar lavage fluid. CMC and OC populations exhibited a protective effect on the allergic asthma rat model as evidenced by means of a partial protection against anaphylactic response and, above all, in the synthesis of IgE and the release of mast cell protease.
Asunto(s)
Antioxidantes/farmacología , Asma/prevención & control , Cacao/química , Polifenoles/farmacología , Sustancias Protectoras/farmacología , Anafilaxia/inducido químicamente , Anafilaxia/prevención & control , Animales , Asma/inducido químicamente , Temperatura Corporal/efectos de los fármacos , Peso Corporal/efectos de los fármacos , Líquido del Lavado Bronquioalveolar/química , Dieta/métodos , Modelos Animales de Enfermedad , Hipersensibilidad/prevención & control , Inmunoglobulina E/análisis , Leucotrienos/análisis , Actividad Motora/efectos de los fármacos , Péptido Hidrolasas/análisis , Perú , RatasRESUMEN
Cigarette smoke is a complex mixture capable of triggering inflammation and oxidative damage in animals at pulmonary and systemic levels. Tempol (4-hydroxy-2,2,6,6-tetramethylpiperidine-1-oxyl) reduces tissue injury associated with inflammation in vivo by mechanisms that are not completely understood. Here we evaluated the effect of tempol on inflammation and oxidative damage induced by acute exposure to cigarette smoke in vivo. Male C57BL/6 mice (n = 32) were divided into 4 groups (n = 8 each): 1) control group exposed to ambient air (GC), 2) animals exposed to cigarette smoke for 5 days (CSG), mice treated 3) prior or 4) concomitantly with tempol (50 mg/kg/day) and exposed to cigarette smoke for 5 days. The results showed that the total number of leukocytes and neutrophils increased in the respiratory tract and lung parenchyma of mice exposed to cigarette smoke. Likewise, MPO levels and activity as well as lipid peroxidation and lung protein nitration and carbonylation also increased. Administration of tempol before or during exposure to cigarette smoke inhibited all the above parameters. Tempol also reduced the pulmonary expression of the inflammatory cytokines Il-6, Il-1ß and Il-17 to basal levels and of Tnf-α by approximately 50%. In contrast, tempol restored Il-10 and Tgf-ß levels and enhanced the expression of Nrf2-associated genes, such as Ho-1 and Gpx2. Accordingly, total GPx activity increased in lung homogenates of tempol-treated animals. Taken together, our results show that tempol protects mouse lungs from inflammation and oxidative damage resulting from exposure to cigarette smoke, likely through reduction of leukocyte infiltration and increased transcription of some of the Nrf2-controlled genes.
Asunto(s)
Óxidos N-Cíclicos/farmacología , Regulación de la Expresión Génica/efectos de los fármacos , Factor 2 Relacionado con NF-E2/metabolismo , Infiltración Neutrófila/efectos de los fármacos , Estrés Oxidativo/efectos de los fármacos , Fumar/efectos adversos , Animales , Líquido del Lavado Bronquioalveolar/química , Interleucina-10/genética , Interleucina-10/metabolismo , Peroxidación de Lípido/efectos de los fármacos , Pulmón/efectos de los fármacos , Pulmón/metabolismo , Pulmón/patología , Masculino , Ratones , Ratones Endogámicos C57BL , Factor 2 Relacionado con NF-E2/genética , Nitritos/análisis , Peroxidasa/metabolismo , Carbonilación Proteica/efectos de los fármacos , Marcadores de Spin , Factor de Crecimiento Transformador beta/genética , Factor de Crecimiento Transformador beta/metabolismoRESUMEN
BACKGROUND: Malaria-triggered lung injury can occur in both severe and non-severe cases. Platelets may interact with parasitized erythrocytes, leukocytes and endothelium. These interactions can lead to microvessel obstructions and induce release of inflammatory mediators. Induction of the haem oxygenase enzyme is important in the host's response to free haem and to several other molecules generated by infectious or non-infectious diseases. In addition, an important role for the haem oxygenase-1 isotype has been demonstrated in experimental cerebral malaria and in clinical cases. Therefore, the present work aims to determine the influence of haem oxygenase in thrombocytopaenia and acute pulmonary injury during infection with Plasmodium berghei strain NK65. METHODS: C57BL/6 mice were infected with P. berghei and analysed 7-10 days post-infection. For each experiment, Cobalt Protoporphyrin IX/CoPPIX or saline were administered. Bronchoalveolar lavage fluid was used for total and differential leukocyte count and for protein measurement. Lungs were used for histological analyses or for analysis of cytokines and western blotting. The lung permeability was analysed by Evans blue dye concentration. Platelet-leukocyte aggregate formation was assayed using the flow cytometer. RESULTS: Plasmodium berghei NK65 infection generated an intense lung injury, with increased levels of inflammatory mediators, oedema, and cell migration into the lung. Plasmodium berghei infection was also accompanied by marked thrombocytopaenia and formation of platelet-leukocyte aggregates in peripheral blood. Treatment with the HO-1 inducer cobalt protoporphyrin IX (CoPPIX) modified the inflammatory response but did not affect the evolution of parasitaemia. Animals treated with CoPPIX showed an improvement in lung injury, with decreased inflammatory infiltrate in the lung parenchyma, oedema and reduced thrombocytopaenia. CONCLUSION: Data here presented suggest that treatment with CoPPIX inducer leads to less severe pulmonary lung injury and thrombocytopaenia during malaria infection, thus increasing animal survival.
Asunto(s)
Lesión Pulmonar Aguda/tratamiento farmacológico , Hemo-Oxigenasa 1/farmacología , Malaria/complicaciones , Proteínas de la Membrana/farmacología , Sustancias Protectoras/farmacología , Trombocitopenia/tratamiento farmacológico , Lesión Pulmonar Aguda/etiología , Animales , Líquido del Lavado Bronquioalveolar/química , Femenino , Recuento de Leucocitos , Pulmón/patología , Masculino , Ratones , Ratones Endogámicos C57BL , Plasmodium berghei/fisiología , Trombocitopenia/etiologíaRESUMEN
The current pandemic of coronavirus disease 19 (COVID-19) has affected millions of individuals and caused thousands of deaths worldwide. The pathophysiology of the disease is complex and mostly unknown. Therefore, identifying the molecular mechanisms that promote progression of the disease is critical to overcome this pandemic. To address such issues, recent studies have reported transcriptomic profiles of cells, tissues and fluids from COVID-19 patients that mainly demonstrated activation of humoral immunity, dysregulated type I and III interferon expression, intense innate immune responses and inflammatory signaling. Here, we provide novel perspectives on the pathophysiology of COVID-19 using robust functional approaches to analyze public transcriptome datasets. In addition, we compared the transcriptional signature of COVID-19 patients with individuals infected with SARS-CoV-1 and Influenza A (IAV) viruses. We identified a core transcriptional signature induced by the respiratory viruses in peripheral leukocytes, whereas the absence of significant type I interferon/antiviral responses characterized SARS-CoV-2 infection. We also identified the higher expression of genes involved in metabolic pathways including heme biosynthesis, oxidative phosphorylation and tryptophan metabolism. A BTM-driven meta-analysis of bronchoalveolar lavage fluid (BALF) from COVID-19 patients showed significant enrichment for neutrophils and chemokines, which were also significant in data from lung tissue of one deceased COVID-19 patient. Importantly, our results indicate higher expression of genes related to oxidative phosphorylation both in peripheral mononuclear leukocytes and BALF, suggesting a critical role for mitochondrial activity during SARS-CoV-2 infection. Collectively, these data point for immunopathological features and targets that can be therapeutically exploited to control COVID-19.
Asunto(s)
Betacoronavirus/inmunología , Quimiocinas/sangre , Infecciones por Coronavirus/inmunología , Interferón Tipo I/sangre , Neutrófilos/inmunología , Neumonía Viral/inmunología , Líquido del Lavado Bronquioalveolar/química , Líquido del Lavado Bronquioalveolar/citología , COVID-19 , Infecciones por Coronavirus/patología , Perfilación de la Expresión Génica , Humanos , Inflamación/virología , Gripe Humana/inmunología , Interferón Tipo I/inmunología , Neutrófilos/citología , Fosforilación Oxidativa , Pandemias , Neumonía Viral/patología , SARS-CoV-2 , Transcriptoma/genéticaRESUMEN
Ha habido mucha discusión sobre los efectos dañinos para la salud producidos por los cigarrillos electrónicos o vapeadores y su utilidad como ayuda para dejar de fumar. Cada vez aparecen más publicaciones con efectos deletéreos sobre la salud. Esta discusión se ha acentuado en los últimos años, por el importante aumento del uso de los vapeadores en todo el mundo, especialmente entre los adolescentes y adultos jóvenes. En septiembre de 2019 el Centro de Control y Prevención de Enfermedades (CDC) de los EE. UU. alertó sobre un importante número de casos de enfermedad pulmonar asociada al uso de cigarrillo electrónico (EVALI: e-cigarette or vaping associated lung injury). Epidemiológicamente se consideró un brote que al 17 de enero, 2020 ha presentado 2.668 pacientes hospitalizados, con 57 fallecidos. Durante la semana del 15 de septiembre 2019 ocurrió el 'peak' de hospitalizaciones por EVALI. La mayoría eran varones jóvenes. El 82% usó productos con Tetrahidrocanabinoides (THC) y el 14% productos con nicotina. En el lavado bronquio-alveolar de 51 casos de EVALI se encontró la presencia de acetato de Vitamina E, producto utilizado como espesante para la elaboración de productos de 'vapeo' que contienen THC, lo que lo hace un posible factor causal, pero no se puede descartar el papel de otros compuestos tóxicos. Las principales sociedades científicas del mundo y la OMS han advertido de los riesgos a largo plazo del uso de los cigarrillos electrónicos y recomiendan su control y regulación.
There has been a lot of discussion about the harmful health effects caused by electronic cigarettes or vapers and their usefulness as a smoking cessation aid. More and more publications appear with deleterious effects on health. This discussion has been straightened in recent years, due to the significant increase in the use of vapers worldwide, especially among adolescents and young adults. In September 2019, the US Center for Disease Control and Prevention warned of a significant number of cases of lung disease associated with the use of electronic cigarettes (EVALI: e-cigarette or vaping associated lung injury). Epidemiologically it was considered an outbreak that as of January 17, 2020 presented 2668 hospitalized patients, with 57 deaths. During the week of September 15, 2019 the peak of hospitalizations for EVALI occurred. The majority were young men. 82% of them used products with Tetrahydrocanabinoids (THC) and 14% products with nicotine. In the bronchoalveolar lavage of 51 cases of EVALI, the presence of Vitamin E acetate was found, a product used as a thickener for the elaboration of vaping products containing THC, which makes it a possible causal factor, but it cannot be ruled out the contribution of other toxic compounds. The world's leading scientific societies and World Health Organization have warned of the long-term risks of using electronic cigarettes and recommend their control and regulation.
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Humanos , Lesión Pulmonar/etiología , Sistemas Electrónicos de Liberación de Nicotina , Vapeo/efectos adversos , Dronabinol , Vitamina E/análisis , Líquido del Lavado Bronquioalveolar/química , Lesión Pulmonar/patología , Lesión Pulmonar/epidemiología , Lesión Pulmonar/diagnóstico por imagenRESUMEN
Idiopathic pulmonary fibrosis (IPF) is a progressive and chronic inflammatory disease with a poor prognosis and very few available treatment options. Low-level laser therapy (LLLT) has been gaining prominence as a new and effective anti-inflammatory and immunomodulatory agent. Can lung inflammation and the airway remodeling be regulated by LLLT in an experimental model of IPF in C57Bl/6 mice? The present study investigated if laser attenuates cellular migration to the lungs, the airway remodeling as well as pro-fibrotic cytokines secretion from type II pneumocytes and fibroblasts. Mice were irradiated (780 nm and 30 mW) and then euthanized fifteen days after bleomycin-induced lung fibrosis. Lung inflammation and airway remodeling were evaluated through leukocyte counting in bronchoalveolar lavage fluid (BALF) and analysis of collagen in lung, respectively. Inflammatory cells in blood were also measured. For in vitro assays, bleomycin-activated fibroblasts and type II pneumocytes were irradiated with laser. The pro- and anti-inflammatory cytokines level in BALF as well as cells supernatant were measured by ELISA, and the TGFß in lung was evaluated by flow cytometry. Lung histology was used to analyze collagen fibers around the airways. LLLT reduced both migration of inflammatory cells and deposition of collagen fibers in the lungs. In addition, LLLT downregulated pro-inflammatory cytokines and upregulated the IL-10 secretion from fibroblasts and pneumocytes. Laser therapy greatly reduced total lung TGFß. Systemically, LLLT also reduced the inflammatory cells counted in blood. There is no statistical difference in inflammatory parameters studied between mice of the basal group and the laser-treated mice. Data obtained indicate that laser effectively attenuates the lung inflammation, and the airway remodeling in experimental pulmonary fibrosis is driven to restore the balance between the pro- and anti-inflammatory cytokines in lung and inhibit the pro-fibrotic cytokines secretion from fibroblasts.
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Remodelación de las Vías Aéreas (Respiratorias) , Citocinas/metabolismo , Fibrosis Pulmonar Idiopática/radioterapia , Rayos Láser , Animales , Líquido del Lavado Bronquioalveolar/química , Células Cultivadas , Citocinas/análisis , Modelos Animales de Enfermedad , Regulación hacia Abajo/efectos de la radiación , Células Epiteliales/citología , Células Epiteliales/metabolismo , Células Epiteliales/efectos de la radiación , Fibroblastos/citología , Fibroblastos/metabolismo , Fibroblastos/efectos de la radiación , Fibrosis Pulmonar Idiopática/patología , Terapia por Láser , Masculino , Ratones , Ratones Endogámicos C57BL , Regulación hacia Arriba/efectos de la radiaciónRESUMEN
Pulmonary tuberculosis (PTB) has been identified as a substantial public health threat and diagnostic challenge. A large proportion of patients exhibit negative smear tests despite active infection. The role of cytokines in the pathophysiology and clinical severity of PTB remains a controversial question. We evaluated the pattern of cytokines presents locally in patients with smear-negative PTB. Levels of tumor necrosis factor (TNF)-α, interferon (IFN)-γ, interleukin (IL)-2, IL-4, IL-6, IL-10, and IL-17 in bronchoalveolar lavage fluid (BALf) from patients with smear-negative PTB, as well as in those with other pulmonary diseases and controls, were performed by flow cytometry. ROC curve and a radiological severity scale were used to establish the potential diagnosis use and the relationship of the cytokine levels with disease severity, respectively. The levels of IL-6 were higher in the PTB (P = 0.0249) and pneumonia (P = 0.0047) groups compared to controls. Low to undetectable levels of TNF-α, IFN-γ, IL-2, IL-4, IL-10, and IL-17 were found in BALf, even after sample concentration using filtration columns and centrifugation. IL-6 levels measured in BALf could distinguish PTB patients or pneumonia patients from controls (AUC: 0.91, P = 0.002 and AUC: 0.86, P = 0.001, respectively), but not patients with PTB from those with pneumonia (AUC: 0.51, P = 0.86). IL-6 levels were related with the severity of PTB, as levels were higher in patients with higher radiological severity. These results confirm the importance of IL-6 in the immunopathology of smear-negative PTB.
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Interleucina-6/metabolismo , Tuberculosis Pulmonar/metabolismo , Tuberculosis Pulmonar/patología , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Líquido del Lavado Bronquioalveolar/química , Femenino , Humanos , Interferón gamma/metabolismo , Masculino , Persona de Mediana Edad , Radiografía/métodos , Tuberculosis Pulmonar/diagnóstico , Factor de Necrosis Tumoral alfa/metabolismo , Adulto JovenRESUMEN
PURPOSE: To investigate the role of vagus nerve activation in the protective effects of hypercapnia in ventilator-induced lung injury (VILI) rats. METHODS: Male Sprague-Dawley rats were randomized to either high-tidal volume or low-tidal volume ventilation (control) and monitored for 4h. The high-tidal volume group was further divided into either a vagotomy or sham-operated group and each surgery group was further divided into two subgroups: normocapnia and hypercapnia. Injuries were assessed hourly through hemodynamics, respiratory mechanics and gas exchange. Protein concentration, cell count and cytokines (TNF-α and IL-8) in bronchoalveolar lavage fluid (BALF), lung wet-to-dry weight and pathological changes were examined. Vagus nerve activity was recorded for 1h. RESULTS: Compared to the control group, injurious ventilation resulted in a decrease in PaO2/FiO2 and greater lung static compliance, MPO activity, enhanced BALF cytokines, protein concentration, cell count, and histology injury score. Conversely, hypercapnia significantly improved VILI by decreasing the above injury parameters. However, vagotomy abolished the protective effect of hypercapnia on VILI. In addition, hypercapnia enhanced efferent vagus nerve activity compared to normocapnia. CONCLUSION: These results indicate that the vagus nerve plays an important role in mediating the anti-inflammatory effect of hypercapnia on VILI.
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Líquido del Lavado Bronquioalveolar/química , Hipercapnia , Nervio Vago/cirugía , Lesión Pulmonar Inducida por Ventilación Mecánica/prevención & control , Animales , Citocinas/análisis , Modelos Animales de Enfermedad , Interleucina-8/análisis , Masculino , Distribución Aleatoria , Ratas , Ratas Sprague-Dawley , Factor de Necrosis Tumoral alfa/análisis , VagotomíaRESUMEN
INTRODUCTION: Idiopathic pulmonary fibrosis (IPF) is a lethal disease with an unclear pathogenic mechanism. Components of the renin-angiotensin system (RAS) have a role in the pathogenesis of IPF, specifically, the aspartyl protease renin acts as a profibrotic factor in the lung. However, the concentration of the RAS components renin and soluble (pro)renin receptor (sPRR) have not been previously evaluated neither in serum nor in bronchoalveolar lavage fluid (BAL) of patients with IPF or chronic Hypersensitivity pneumonitis (cHP), a disease which may be confused with IPF. METHODS: The serum levels of renin [IPF patients (n = 70), cHP patients (n = 83), and controls (n = 26)] and sPRR [IPF (n = 28), cHP (37), and controls (n = 20)] were measured by ELISA. Renin was also quantified in BALs of IPF patients and controls by Western blot. RESULTS: We found that the levels of renin were higher in serum samples from IPF patients when compared with cHP patients and controls. Furthermore, BALs from IPF patients had more renin than BALs from controls. Unlike renin, the serum levels of sPRR were lower in IPF and cHP patients than in control individuals. CONCLUSIONS: The high levels of renin in sera and BALs of IPF patients suggest that renin might play a major role in the pathogenesis of IPF. Results from BAL confirm that renin is produced locally in the lung. Serum levels of renin could be used to differentiate IPF from cHP.
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Alveolitis Alérgica Extrínseca/metabolismo , Fibrosis Pulmonar Idiopática/metabolismo , Receptores de Superficie Celular/sangre , Renina/metabolismo , ATPasas de Translocación de Protón Vacuolares/sangre , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Alveolitis Alérgica Extrínseca/fisiopatología , Western Blotting , Líquido del Lavado Bronquioalveolar/química , Enfermedad Crónica , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , Fibrosis Pulmonar Idiopática/fisiopatología , Masculino , Persona de Mediana Edad , Oxígeno/metabolismo , Capacidad de Difusión Pulmonar , Capacidad Vital , Adulto JovenRESUMEN
INTRODUCTION: The evaluation of expectorant activity has been extensively studied in murine models, involving the secretion of phenol red in the trachea or bronchus to estimate the secretory capacity of lower airway mucosa. However, differences in the experimental protocols of several studies evidenced the need of to standardize the quantification of phenol red in the bronchoalveolar fluid (BALF). METHODS: The analytical methodology for the quantification of phenol red in the BALF was optimized by investigation of pH influence, quantity of the alkali agent added and appropriate wavelength for quantification of phenol red by UV-VIS spectroscopy. Different phenol red suspensions (0.05, 0.5, 1.25, 2.5 and 5%) were prepared and administered intraperitoneally in mice at doses 5, 25, 50, 250 or 500â¯mg/kg. RESULTS: It was shown that phenol red should be used at dose 500â¯mg/kg and intraperitoneal administration should be performed from a suspension at 1.25% (w/v). Furthermore, the alkalinizing agent of choice would be NaOH (0.1â¯M). The pharmacological validation of the analytical method showed that ambroxol (30, 60 or 120â¯mg/kg), guaifenesin (100â¯mg/kg), NH4Cl (2000â¯mg/kg) or salbutamol (4â¯mg/kg) can be used as positive controls. DISCUSSION: The phenol red quantification in the BALF is a rapid and low cost assay for the discovery of new expectorant drugs. Thus, it was proposed a standardization of the analytical and pharmacological methods to ensure the reliability of BALF processing and reproducibility of phenol red quantification for data analysis.
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Expectorantes/química , Fenolsulfonftaleína/química , Animales , Antiácidos/química , Bronquios/química , Líquido del Lavado Bronquioalveolar/química , Masculino , Ratones , Modelos Animales , Preparaciones Farmacéuticas/química , Reproducibilidad de los Resultados , Tráquea/químicaRESUMEN
Glucagon has been shown to be beneficial as a treatment for bronchospasm in asthmatics. Here, we investigate if glucagon would prevent airway hyperreactivity (AHR), lung inflammation, and remodeling in a murine model of asthma. Glucagon (10 and 100 µg/Kg, i.n.) significantly prevented AHR and eosinophilia in BAL and peribronchiolar region induced by ovalbumin (OVA) challenge, while only the dose of 100 µg/Kg of glucagon inhibited subepithelial fibrosis and T lymphocytes accumulation in BAL and lung. The inhibitory action of glucagon occurred in parallel with reduction of OVA-induced generation of IL-4, IL-5, IL-13, TNF-α, eotaxin-1/CCL11, and eotaxin-2/CCL24 but not MDC/CCL22 and TARC/CCL17. The inhibitory effect of glucagon (100 µg/Kg, i.n.) on OVA-induced AHR and collagen deposition was reversed by pre-treatment with indomethacin (10 mg/Kg, i.p.). Glucagon increased intracellular cAMP levels and inhibits anti-CD3 plus anti-CD28-induced proliferation and production of IL-2, IL-4, IL-10, and TNF- α from TCD4+ cells in vitro. These findings suggest that glucagon reduces crucial features of asthma, including AHR, lung inflammation, and remodeling, in a mechanism probably associated with inhibition of eosinophils accumulation and TCD4+ cell proliferation and function. Glucagon should be further investigated as an option for asthma therapy.