Bronchoalveolar Lavage Fluid Supernatant Research Articles

Biophysical properties of alveolar surfactant in drever dogs with hunting associated pulmonary edema

BackgroundA syndrome of acute non-cardiogenic pulmonary edema associated with hunting is prevalent in the drever breed, but etiology of this syndrome is currently unknown. Alveolar surfactant has a critical role in preventing alveolar collapse and edema formation. The aim of this study was to investigate, whether the predisposition to hunting associated pulmonary edema in drever dogs is associated with impaired biophysical properties of alveolar surfactant. Seven privately owned drever dogs with recurrent hunting associated pulmonary edema and seven healthy control dogs of other breeds were included in the study. All affected dogs underwent thorough clinical examinations including echocardiography, laryngeal evaluation, bronchoscopy, and bronchoalveolar lavage (BAL) as well as head, neck and thoracic computed tomography imaging to rule out other cardiorespiratory diseases potentially causing the clinical signs. Alveolar surfactant was isolated from frozen, cell-free supernatants of BAL fluid and biophysical analysis of the samples was completed using a constrained sessile drop surfactometer. Statistical comparisons over consecutive compression expansion cycles were performed using repeated measures ANOVA and comparisons of single values between groups were analyzed using T-test.ResultsThere were no significant differences between groups in any of the biophysical outcomes of surfactant analysis. The critical function of surfactant, reducing the surface tension to low values upon compression, was similar between healthy dogs and affected drevers.ConclusionsThe etiology of hunting associated pulmonary edema in drever dogs is not due to an underlying surfactant dysfunction.

Alzheimer’s disease induces pathological changes in the lungs of humans and the AppNL‐G‐F mouse model

AbstractBackgroundAlzheimer’s disease (AD) is an age‐related neurodegenerative disorder characterized by brain accumulation of amyloid beta (Aβ) plaques, neurofibrillary tangles, neuron death, robust gliosis, and neuroinflammation, affecting around 35 million individuals worldwide. It is well known that brain immune changes are a component of the disease, but immune dysfunction in secondary organs has also been reported. To assess peripheral immune changes in AD, we asked whether lung dysfunction and inflammatory change are present during the disease.MethodMale and female C57BL/6 wild type and AppNL‐G‐F mice (8‐9 months old) were sacrificed, and broncho‐alveolar lavage fluid (BALF) and lungs were collected. BALF was used to quantify immune cell phenotype by flow cytometry, and the BALF supernatant was used to determine Aβ and cytokine concentrations and total protein content. Lungs were sectioned and stained with hematoxylin and eosin, Alcian blue and Masson’s trichrome. In addition, lung sections from wild‐type and AppNL‐G‐F mice and human AD patient and healthy controls were immunostained for APP, Aβ and claudin 4.ResultAD mice showed significantly increased pulmonary airway thickening and mucus and collagen production as indicated by hematoxylin and eosin, Alcian blue and Masson’s trichrome staining, respectively, compared to the wild‐type controls. Female but not male AppNL‐G‐F mice also demonstrated reduced B cell and neutrophil numbers and elevated IL‐1β in their BALF compared to controls. Although there were no differences in BALF supernatant total protein content between wild‐type and AppNL‐G‐F mice, lungs from both sexes of AppNL‐G‐F mice had decreased claudin‐4 immunoreactivity suggesting altered epithelial permeability. BALF supernatant from male and female AppNL‐G‐F mice also contained soluble Aβ1‐40 and Aβ1‐42 correlating with lung APP immunoreactivity. Human AD male and female lungs had reduced claudin‐4 immunoreactivity and increased APP and Aβ staining compared to controls verifying lung changes are also present in human disease.ConclusionThese data demonstrate disease‐associated changes in the lungs of both human AD patients and the AppNL‐G‐F mouse model. Further characterization of lung dysfunction and its contribution to brain changes may define a novel lung‐to‐brain contribution to disease.

A Murine Model of Vesicant-Induced Acute Lung Injury.

Burn injuries including those caused by chemicals can result in systemic effects and acute lung injury (ALI). Cutaneous exposure to Lewisite, a warfare and chemical burn agent, also causes ALI. To overcome the limitations in conducting direct research on Lewisite-induced ALI in a laboratory setting, an animal model was developed using phenylarsine oxide (PAO) as a surrogate for Lewisite. Due to lack of a reliable animal model mimicking the effects of such exposures, development of effective therapies to treat such injuries is challenging. We demonstrated that a single cutaneous exposure to PAO resulted in disruption of the alveolar-capillary barrier as evidenced by elevated protein levels in the bronchoalveolar lavage fluid (BALF). BALF supernatant of PAO-exposed animals had increased levels of high mobility group box 1, a damage associated molecular pattern molecule. Arterial blood-gas measurements showed decreased pH, increased PaCO2, and decreased partial pressure of arterial O2, indicative of respiratory acidosis, hypercapnia, and hypoxemia. Increased protein levels of interleukin (IL)-6, CXCL-1, CXCL-2, CXCL-5, granulocyte-macrophage colony-stimulating factor, CXCL-10, leukemia inhibitory factor, leptin, IL-18, CCL-2, CCL-3, and CCL-7 were observed in the lung of PAO-exposed mice. Further, vascular endothelial growth factor levels were reduced in the lung. Pulmonary function evaluated using a flexiVent showed a downward shift in the pressure-volume loop, decreases in static compliance and inspiratory capacity, increases in respiratory elastance and tissue elastance. These changes are consistent with an ALI phenotype. These results demonstrate that cutaneous PAO exposure leads to ALI and that the model can be used as an effective surrogate to investigate vesicant-induced ALI. SIGNIFICANCE STATEMENT: This study presents a robust model for studying ALI resulting from cutaneous exposure to PAO, a surrogate for the toxic vesicating agent Lewisite. The findings in this study mimic the effects of cutaneous Lewisite exposure, providing a reliable model for investigating mechanisms underlying toxicity. The model can also be used to develop medical countermeasures to mitigate ALI associated with cutaneous Lewisite exposure.

Ophiopogonin A attenuates acute lung inflammation in Klebsiella pneumonia mice by regulating RELA (v-rel reticuloendotheliosis viral oncogene homolog A (avian))

Ophiopogonin are natural products from the roots of Ophiopogon japonicas with various pharmacological activities. This study explored the mechanism of ophiopogonin A to alleviate the acute inflammation of the lungs of Klebsiella pneumonia. 64 rats were equally assigned into blank control group, injury model group (intratracheal injection of Klebsiella pneumonia to establish Klebsiella pneumonia model), Ophiopogonin A group and Ophiopogonin A+RELA group (combined group, on the basis of injury model group, received RELA mediated ophiopogonin A), with the dose of 10 mg/kg. The histopathological damage was observed under the microscope after HE staining, as well as inflammatory cells in bronchoalveolar lavage fluid (BALF), cytokine levels, and expression of RELA. After HE staining, compared with injury model group, bronchial epithelium structure in Ophiopogonin A group and combination group were relatively intact, the degree of infiltration of inflammatory cells was reduced, and the thickness of alveolar septum was reduced, especially in combination group. The pathological scores of Ophiopogonin A group and combination group were significantly lower, with a further lower score in combination group. The number of inflammatory cells in BALF of rats in Ophiopogonin A group and combination group were significantly reduced, especially in combination group. Similarly, cytokine levels in BALF supernatant and serum of rats in treatment group were reduced, especially in combination group (P < 0.05). Ophiopogonin A could down-regulate RELA, which was further reduced in combination group. In conclusion, ophiopogonin A can reduce the invasion and pathological damage of pathogens to lung tissue and exert an anti-inflammatory ability.

Fatty acid fingerprints in bronchoalveolar lavage fluid and its extracellular vesicles reflect equine asthma severity

Equine asthma (EA) is an inflammatory disease of the lower airways driven by mediators released from cells. Extracellular vesicles (EVs) are vehicles for lipid mediators, which possess either pro-inflammatory or dual anti-inflammatory and pro-resolving functions. In this study, we investigated how the respiratory fatty acid (FA) profile reflects airway inflammatory status. The FA composition of bronchoalveolar lavage fluid (BALF), BALF supernatant, and bronchoalveolar EVs of healthy horses (n = 15) and horses with mild/moderate EA (n = 10) or severe EA (SEA, n = 5) was determined with gas chromatography and mass spectrometry. The FA profiles distinguished samples with different diagnoses in all sample types, yet they were insufficient to predict the health status of uncategorized samples. Different individual FAs were responsible for the discrimination of the diagnoses in different sample types. Particularly, in the EVs of SEA horses the proportions of palmitic acid (16:0) decreased and those of eicosapentaenoic acid (20:5n-3) increased, and all sample types of asthmatic horses had elevated dihomo-γ-linolenic acid (20:3n-6) proportions. The results suggest simultaneous pro-inflammatory and resolving actions of FAs and a potential role for EVs as vehicles for lipid mediators in asthma pathogenesis. EV lipid manifestations of EA can offer translational targets to study asthma pathophysiology and treatment options.

Analysis of cytokines in bronchoalveolar lavage fluid in patients with checkpoint inhibitor pneumonitis and pulmonary infection: A case-control study.

The discovery of immune checkpoint inhibitors (ICIs) is a breakthrough in the field of cancer therapy. However, ICIs may cause immune-related adverse reactions, including checkpoint inhibitor-related pneumonitis (CIP). The aim of this study was to investigate cytokines in bronchoalveolar lavage fluid (BALF) of patients with CIP compared with patients with pulmonary infection and patients with cancer. We retrospectively analyzed 34 cytokine levels and T cell subsets in BALF supernatant samples from ICI-treated patients with CIP (n = 13), pulmonary infection (n = 10), and progressive cancer (n = 12). Cytokine levels and T cell subsets were compared among the three groups of patients. We observed significantly higher levels of IFN-γ-induced protein 10(IP-10) (p = 0.002), and percentage of CD3 + CD8 + T cells (p = 0.020) in BALF of patients with CIP compared with the other two groups. However, we found significantly lower levels of interleukin-21 (p = 0.008) in BALF of patients with progressive disease compared with the other two groups. Cytokine profile and character of cell subsets in BALF was helpful for the differential diagnosis of CIP. IP-10 may play an important role in pathophysiology for CIP and also be a potential therapeutic target.

Effects and mechanism of annexin A1-overexpressing human adipose-derived mesenchymal stem cells in the treatment of mice with acute respiratory distress syndrome

Effects and mechanism of annexin A1-overexpressing human adipose-derived mesenchymal stem cells in the treatment of mice with acute respiratory distress syndrome

Pulmonary pathogenesis in a murine model of inhaled arsenical exposure.

Arsenic trioxide (ATO), an inorganic arsenical, is a toxic environmental contaminant. It is also a widely used chemical with industrial and medicinal uses. Significant public health risk exists from its intentional or accidental exposure. The pulmonary pathology of acute high dose exposure is not well defined. We developed and characterized a murine model of a single inhaled exposure to ATO, which was evaluated 24h post-exposure. ATO caused hypoxemia as demonstrated by arterial blood-gas measurements. ATO administration caused disruption of alveolar-capillary membrane as shown by increase in total protein and IgM in the bronchoalveolar lavage fluid (BALF) supernatant and an onset of pulmonary edema. BALF of ATO-exposed mice had increased HMGB1, a damage-associated molecular pattern (DAMP) molecule, and differential cell counts revealed increased neutrophils. BALF supernatant also showed an increase in protein levels of eotaxin/CCL-11 and MCP-3/CCL-7 and a reduction in IL-10, IL-19, IFN-γ, and IL-2. In the lung of ATO-exposed mice, increased protein levels of G-CSF, CXCL-5, and CCL-11 were noted. Increased mRNA levels of TNF-a, and CCL2 in ATO-challenged lungs further supported an inflammatory pathogenesis. Neutrophils were increased in the blood of ATO-exposed animals. Pulmonary function was also evaluated using flexiVent. Consistent with an acute lung injury phenotype, respiratory and lung elastance showed significant increase in ATO-exposed mice. PV loops showed a downward shift and a decrease in inspiratory capacity in the ATO mice. Flow-volume curves showed a decrease in FEV0.1 and FEF50. These results demonstrate that inhaled ATO leads to pulmonary damage and characteristic dysfunctions resembling ARDS in humans.

Tools for assessing lung fluid in neonates with respiratory distress

BackgroundTransient tachypnea of the newborn (TTN), as a common cause of neonatal respiratory distress, needs to be distinguished from respiratory distress syndrome (RDS). Various modalities such as lung ultrasonography, cytokine analysis, and electrical cardiometry for the evaluation of lung fluid can be helpful for the exact diagnosis, however, clinical diagnosis has been applied mainly. This study aimed to evaluate the usefulness of the various tools for the diagnosis of TTN and RDS in neonates.MethodsThis study evaluated 22 late-preterm and term infants admitted to the neonatal intensive care unit of Gangnam Severance Hospital because of respiratory distress. Total 9 neonates were diagnosed with TTN and 13 had RDS. In addition to chest radiography, the LUS score was calculated by a neonatologist using the portable ultrasound device. Cytokines in the bronchoalveolar lavage fluid supernatant were measured. Thoracic fluid content was measured using an electrical cardiometry device.ResultsWe enrolled 22 patients with median gestational age, 37.1 weeks, and birth weight 3100 g. There is no difference in patient characteristics between RDS and TTN group. Lung ultrasound score was significantly higher in RDS than TTN (11 vs 6, p = 0.001). Score 0 is shown in all infants with TTN. Score 1 is shown as significantly more in RDS than TTN. Between the TTN and RDS groups, there were significant differences in the changes of thoracic fluid content (2 vs − 1.5, p < 0.001), IL-1β levels (2.5 vs 11.3, p = 0.02), and TNF-α levels (20.1 vs 11.2, p = 0.04).ConclusionWe found lung ultrasound and electrical cardiometry to be reliable diagnostic tools for assessing infants with respiratory distress among late-preterm and term infants. Further studies with a large number of patients are needed to confirm their clinical usefulness.

The influence of a biomimetic pulmonary surfactant modification on the in vivo fate of nanoparticles in the lung

Direct biomimetic modification of nanoparticles (NPs) with endogenous surfactants is helpful to improve the biocompatibility of NPs and avoid damage to the physiological function of the lung. Therefore, the objective of this study is to investigate the influence of biomimetic endogenous pulmonary surfactant phospholipid modification on the in vivo fate of NPs after lung delivery. Here, two neutral phospholipids (dipalmitoylphosphatidylcholine (DPPC), dipalmitoylphosphatidylamine (DPPE)) and two negatively charged phospholipids (dipalmitoylphosphatidylglycerol (DPPG), dipalmitoylphosphatidylserine (DPPS)) were selected to modify paclitaxel (PTX)-loaded PLGA NPs with different molar ratio. DPPC, DPPE, and DPPG improved mucoadhesion, in contrast, DPPS improved the mucus permeability of the NPs. Neutral DPPC and DPPE reduced, but negatively charged DPPS and DPPG increased the uptake by alveolar macrophages, all types of phospholipid increased the uptake by lung epithelial cells and increased PTX retention in the whole lung. Whereas, DPPC, DPPE, and DPPG promoted PTX retention in bronchoalveolar lavage fluid (BALF), while DPPS promoted PTX absorption in the lung tissue. Only DPPS-PLGA (1:1) NPs remarkably increased PTX systemic exposure. A good correlation between PTX percentage in the supernatant of BALF and PTX concentration in plasma was established, implying PTX entered the system circulation mainly in molecular form. Phospholipid modification had no effect on extrapulmonary organ distribution of PTX. Taken together, our study disclosed that different phospholipid modification can endow the NPs mucoadhesive or mucus penetration and cellular uptake properties, with tunable retention in BALF and absorption in the lung tissue, providing the scientific background for translational nanocarrier design for inhalation as required. Statement of significanceInhaled nanomedicines will inevitably interact with pulmonary surfactant and form “surfactant corona”. However, the contribution of individual pulmonary surfactant phospholipid on the in vivo fate of nanomedicines is still unclear. In this regard, the most abundant pulmonary surfactant phospholipid dipalmitoylphosphatidylcholine and dipalmitoylphosphatidylamine, and dipalmitoylphosphatidylglycerol and dipalmitoylphosphatidylserine were selected to modify the paclitaxel loaded PLGA nanoparticles and the effect of these pulmonary surfactant phospholipids on their in vivo fate was investigated. It was demonstrated that different phospholipid modification can endow the nanoparticles mucoadhesive or mucus penetration properties, with tunable retention in bronchoalveolar lavage fluid, alveolar macrophages uptake and absorption in the lung tissue. The present study provided a comprehensive understanding for the role of pulmonary surfactant phospholipid on inhaled nanomedicines.

Clinical value of alveolar lavage supernatant specimens in the detection of the EGFR gene mutation in patients with non-small cell lung carcinoma.

BackgroundThis study sought to compare the consistency of the epidermal growth factor receptor (EGFR) gene mutation detection results in the supernatant of alveolar lavage specimens to the tissue sample results, and the consistency of the blood EGFR gene mutation detection results to the tissue detection results.MethodsIn total, 29 patients with non-small cell lung carcinoma (NSCLC) were selected, and their bronchoalveolar lavage fluid (BALF) was collected. The supernatant and precipitate were separated by centrifugation. Deoxyribonucleic acid (DNA) was extracted from the supernatant, and blood and tumor tissues were collected to detect patients’ EGFR gene mutation status.ResultsOf the 29 enrolled patients, 12 of the 23 tissue-biopsy patients (52.2%) were positive for EGFR mutations, 11 of the 28 blood-test patients (39.2%) were positive for EGFR mutations, and 13 of the 29 cases of the BALF-test patients (44.8%) were positive for EGFR mutations. The most common mutations were the exon 19 deletion mutation and the L858R point mutation. The EGFR gene mutation rate was higher in female, young, non-smoker, and stage IIIB patients (than stage IV patients), but the differences were not statistically significant (all P>0.05). Of the 29 NSCLC patients tested for the EGFR gene mutation, the BALF supernatant and blood results were the same for 27 patients (coincidence rate: 93.10%). Of the 23 of the 29 enrolled patients tested for the EGFR gene mutation, the BALF supernatant and tissue test results were the same for 21 patients (coincidence rate: 91.30%). Further, the blood-test and the tissue test results were the same for 20 patients (coincidence rate: 86.96%).ConclusionsThe EGFR gene mutation rate was high in NSCLC patients. The coincidence rate of the EGFR gene mutation detection results between BALF supernatant and tumor tissues was slightly higher than that of the blood and tumor tissue EGFR gene mutation detection results.

L‐Citrulline Modulates Macrophage Polarization to an M2 Phenotype in a Model of Lipopolysaccharide‐Induced Lung Injury in Neonatal Rats

IntroductionBronchopulmonary dysplasia (BPD), a chronic lung disease in premature infants resulting from inflammation and oxidative stress, leads to arrested pulmonary development and lifelong respiratory morbidities. Macrophages are the most abundant immune cells in the neonatal lung, orchestrating inflammation, maintaining homeostasis, and supporting tissue remodeling and repair. Macrophages can polarize into specific phenotypes in response to the micro‐environment. M1 macrophages are pro‐inflammatory and pathogen killing whereas, M2 macrophages are anti‐inflammatory and support normal lung development. Existing therapies for BPD have inherent side effects. L‐Citrulline (L‐CIT), a non‐essential amino acid, that is safe and well‐tolerated by infants, reduces oxidative stress and lung inflammation. Thus, we hypothesize that L‐CIT modulates macrophage polarization to an M2 phenotype in the presence of lipopolysaccharide (LPS) induced lung injury during early development.MethodsSprague Dawley rat pups received saline (SAL) or L‐CIT (2.5 g/kg) intraperitoneally during postnatal day (PND) 4‐9. In addition, pups were treated with LPS (5mg/kg) or SAL intrapharyngeally during PND 5‐8. Thus, we had 4 treatment groups, ie., SAL+SAL (controls), LPS+SAL, LPS+L‐CIT and L‐CIT+SAL. Pups were euthanized on PND 9. Bronchoalveolar Lavage Fluid (BALF): BALF was cytospinned, and cells were counted following hematoxylin and eosin staining. BALF supernatant was collected, and an arginase activity assay was performed for M2 marker expression. Lung homogenate: Lungs were dissected, homogenized and processed for western blotting of M2 (CD206) and M1 (Arg2) markers normalized to rat macrophage marker (F4/80).ResultsBALF analysis: The number of alveolar macrophages infiltrated in the BALF of animals treated with L‐CIT+LPS (n=4) and those treated with SAL+LPS (n=4) were similar; both were significantly higher than control (p&lt;.001; p&lt;.0001, respectively). Arginase activity in the BALF was significantly higher in animals treated with L‐CIT+LPS (p&lt;.05; n= 4) compared to LPS alone (p&lt;.05; n=4). Lung homogenate analysis: LPS+SAL treated animals had lower levels of protein expression for CD206 (M2 marker) (p&lt;.05; n=4) compared to control animals. Whereas L‐CIT+LPS treated animals had increased expression of CD206 (M2 marker) (p&lt;.05; n=4) and lower expression of Arg2 (M1 marker) (p&lt;.001; n=4) compared to control animals.ConclusionIn BPD pathogenesis, there is a major shift towards M1 polarization due to the presence of pro‐inflammatory stimuli in the microenvironment, leading to arrested lung development. Our preliminary findings support a role for L‐CIT in regulating macrophage polarization, favoring an anti‐inflammatory M2 phenotype in the presence of inflammatory injury. This further suggests that L‐CIT, a safe nutritional supplement, could reduce inflammatory lung injury during early development.

Assessment of cytotoxicity of an original industrial aerosol containing a high percentage of amorphous silica in the nanometer range

Introduction. At the current stage of hygienic evaluation, the substantiation of at least approximate safe concentrations in the ambient air of populated areas for nanoparticles is an up-to-date challenge. Its persistence dissolves the guidelines for risk management and divests the supervisory bodies of legal support. A comparative toxicological evaluation of the studied substance and its chemical analogue is one of the guidelines for the academic substantiation of the hygienic standards for the permissible content of hazardous substances in the air. It already has previously defined exposure standards. Materials and methods. To investigate the cytotoxicity of the studied particles, a shift of the cellular composition of the bronchoalveolar lavage fluid (BALF) was used. Also, some biochemical measurements of the BALF supernatant were investigated. Outbred female rats were instilled with a suspension of particles in the volume of 1 ml of various concentrations in the form of an intratracheal suspension. Distilled water was used as a solvent. Statistical analysis of the data obtained was performed using the Student’s t-test. Results. The comparative assessment of the cytotoxicity of an original industrial aerosol containing 72% amorphous silica with an average particle size of 90 nm (SiO2 IA) was performed. It also included engineered particles of amorphous silica with an average size of 43 nm (SiO2 NPs), a commercial, an industrial sample of 100% amorphous silica with a particle size of 5 to 60 nm (amorphous SiO2), and a reference sample of standard quartz DQ12 in a volume of 1 ml of water suspension. Under the findings of changes in the cellular composition of the bronchoalveolar lavage fluid 24 hours after the intratracheal instillation of these particles, it was revealed that the biological power (in terms of the NL/AM ratio) of both SiO2 NPs and amorphous SiO2 is statistically much higher than the industrial aerosol under study. It is also higher than the standard quartz dust DQ12. In this regard, the cytotoxicity of SiO2 IA may be explained by the predominant content of amorphous silica nanoparticles in it. Conclusion. Under the obtained results, the appropriateness of using indicative safe exposure levels (ISEL) of 0.02 mg/m3 for amorphous silica needs to be reviewed. The safe reference level of impact guideline does not contain data concerning the particle size and the percentage of amorphous SiO2 in the aerosol. Nevertheless, it is impossible to pollute the ambient air with an aerosol containing only this substance.

Anisakis pegreffii Extract Induces Airway Inflammation with Airway Remodeling in a Murine Model System.

Exposure of the respiratory system to the Anisakis pegreffii L3 crude extract (AE) induces airway inflammation; however, the mechanism underlying this inflammatory response remains unknown. AE contains allergens that promote allergic inflammation; exposure to AE may potentially lead to asthma. In this study, we aimed to establish a murine model to assess the effects of AE on characteristic features of chronic asthma, including airway hypersensitivity (AHR), airway inflammation, and airway remodeling. Mice were sensitized for five consecutive days each week for 4 weeks. AHR, lung inflammation, and airway remodeling were evaluated 24 h after the last exposure. Lung inflammation and airway remodeling were assessed from the bronchoalveolar lavage fluid (BALF). To confirm the immune response in the lungs, changes in gene expression in the lung tissue were assessed with reverse transcription-quantitative PCR. The levels of IgE, IgG1, and IgG2a in blood and cytokine levels in the BALF, splenocyte, and lung lymph node (LLN) culture supernatant were measured with ELISA. An increase in AHR was prominently observed in AE-exposed mice. Epithelial proliferation and infiltration of inflammatory cells were observed in the BALF and lung tissue sections. Collagen deposition was detected in lung tissues. AE exposure increased IL-4, IL-5, and IL-13 expression in the lung, as well as the levels of antibodies specific to AE. IL-4, IL-5, and IL-13 were upregulated only in LLN. These findings indicate that an increase in IL-4+ CD4+ T cells in the LLN and splenocyte resulted in increased Th2 response to AE exposure. Exposure of the respiratory system to AE resulted in an increased allergen-induced Th2 inflammatory response and AHR through accumulation of inflammatory and IL-4+ CD4+ T cells and collagen deposition. It was confirmed that A. pegreffii plays an essential role in causing asthma in mouse models and has the potential to cause similar effects in humans.

ILC3-derived acetylcholine promotes protease-driven allergic lung pathology

ILC3-derived acetylcholine promotes protease-driven allergic lung pathology

Effect of TLR7 gene expression mediating NF-κB signaling pathway on the pathogenesis of bronchial asthma in mice and the intervention role of IFN-γ.

To explore the mechanism of TLR7 mediating NF-κB signaling pathway on the pathogenesis of bronchial asthma in mice and the intervention effect of IFN-γ in the process. The experimental animals were 70 C57BL/6J female mice of clean grade, which were divided into 7 groups according to different treatment protocols, including Normal group, Asthma group, Model+1-MT group, Model+IFN-γ group, Model+TLR7 agonist group, TLR7 deficient group, and Model+TLR7 deficient group. Hematoxylin-eosin (HE) staining was used to observe the pathological changes of lung tissues. The positive expression rates of TLR7, p-IKKα and NF-κBp65 were detected by immunohistochemistry. bronchoalveolar lavage fluid (BALF) cells were classified and counted. The contents of interleukin (IL)-4, IL-10, IL-12 and interferon (IFN)-γ in BALF supernatant were detected by enzyme-linked immunosorbent assay (ELISA). Following isolation, culture and plasmid construction of airway smooth muscle cells (ASMCS) from normal mice and asthmatic mice, cells were transfected and divided into the Control group, pcDNA-TLR7 NC group, siRNA-TLR7 NC group, pcDNA-TLR7 group, siRNA-TLR7 group, Asatone group, Triptolide group, and pcDNA-TLR7 +Asatone group. The expression of TLR7, IDO, p-IKKα and NF-κBp65 was detected by real-time polymerase chain reaction (RT-PCR) and Western blot, respectively. 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) was used to detect the proliferation of ASMCS. The cell cycle and apoptosis of ASMCS were detected by flow cytometry. HE staining showed successful modeling of asthma. Immunohistochemical test showed that the positive expression rate of TLR7 in the Asthma group was significantly decreased, and that of IKKα and NF-κBp65 was significantly increased, with significantly increased IL-4, IL-10, IL-12 and IFN-γ levels (all p<0.05). Model+1-MT group and Model+TLR7 deficient group had a large number of inflammatory cell infiltration, increased IL-4, IL-10, IL-12 and IFN-γ levels, decreased expression levels of TLR7 and IDO, and increased expression of p-IKKα and NF-κBp65 (all p<0.05); while the opposites results were detected in Model+IFN-γ group and Model+TLR7 agonist group (all p<0.05). Cell transfection experiments revealed that pcDNA-TLR7 group and Triptolide group had increased TLR7 expression while decreased p-IKKα and NF-κBp65, decreased proliferation level, and increased cell apoptosis (all p<0.05); while the contrary results were found in siRNA-TLR7 group and Asatone group (all p<0.05); yet without significant difference in pcDNA-TLR7+Asatone group (all p>0.05). Upregulation of TLR7 can inhibit the activation of NF-κB signaling pathway, reduces airway inflammation, inhibits ASMCS proliferation and thus promotes cell apoptosis in asthmatic mice. Besides, IFN-γ can exert a protective role in suppressing the progression of inflammation in asthma.

Metabolomics analysis of bronchoalveolar lavage fluid samples in horses with naturally-occurring asthma and experimentally-induced airway inflammation

The present work characterized the metabolomic profile of bronchoalveolar lavage fluid (BALF) in healthy horses, experimentally-induced airway inflammation by lipopolysaccharide (LPS) nebulization, and naturally-occurring asthma (n = 3 in each group). All animals underwent clinical and upper airway endoscopic examinations, and bronchoalveolar lavage. BALF supernatant samples were subjected to metabolic analysis based on gas chromatography–mass spectrometry (GC-MS). Overall, 67 peaks were obtained from BALF GC-MS analysis, corresponding to 53 metabolites which were categorized according to chemical class, such as organic acids, fatty acids, nucleosides or their derivatives, amino acids, peptides or their derivatives, carbohydrates, and other compounds. Our results showed that the airway inflammation induction model with LPS produced the same pattern of metabolite changes as in horses with naturally occurring asthma. Metabolic pathway analysis was done by means of Fisher's exact test, for detection of metabolites over-represented in asthma affected-horses and LPS-induced airway inflammation as compared with healthy horses. The most significant altered metabolic pathways were fatty acid biosynthesis, galactose metabolism and citrate cycle. These results suggest that the airway inflammation induction model with LPS is a good study model for asthma-affected horses, due to the similarity of the profile of inflammatory cells (specifically neutrophils) and similar metabolic alterations found in BALF that occur during the inflammatory process of the airways. Further research may increase understanding of metabolomics disturbances and their significance in the pathogenesis of equine asthma.

Clinical Feasibility and Airway Deposition of Nebulized Voriconazole in Healthy Horses

Voriconazole (VRC) is a potential treatment for pneumomycosis in horses. The objectives of this study were to determine if the delivery of Vfend using a Flexineb nebulizer produced clinically significant [VRC] in lower airways. The hypothesis was that [VRC] after delivery by nebulization would be greater in the pulmonary epithelial lining fluid than plasma. A secondary objective was to determine [VRC] in upper airways through the collection of nasopharyngeal wash (NPW) samples. Voriconazole solution [Vfend-6.25 mg/mL, 100 (n = 2), 200 (n = 3), 500 (n = 1) mg] was nebulized once in 6 healthy geldings. Clinical responses, duration of nebulization, and [VRC] at various time points (up to 8 hours) in plasma, bronchoalveolar lavage fluid (BALF) supernatant and cell pellet, and NPW samples were recorded. Voriconazole (Vfend-6.25 mg/mL, 200 mg) was nebulized in 5 additional, healthy geldings, and [VRC] was measured in NPW samples pre- and postnebulization at time points up to 8 hours. The antifungal activity of BALF and NPW samples was determined using agar disk diffusion. Concentrations of voriconazole were below detection in plasma, BALF supernatant, and cell pellets for all time points and doses except the BALF cell pellet (0.4 μg/g) immediately after nebulization of 500 mg. For 5 horses, administered 200 mg of Vfend, mean [VCR] in NPW at the end of nebulization and 1, 6, and 8 hours postnebulization were: 30.8 ± 29, 1.0 ± 0.84, 0.2 ± 0.19, and 0.34 ± 0.67 μg/mL, respectively. Only NPW samples obtained immediately postnebulization showed antifungal activity. A nebulized Vfend solution is not recommended for the treatment of pneumomycosis in horses.

Application of dexmedetomidine for lung injury in elderly patients undergoing one-lung ventilation.

This study aimed to evaluate the effects of dexmedetomidine (DEX) on lung injury, the oxygenation index and perioperative pulmonary complications in elderly patients who underwent thoracotomy with one-lung ventilation (OLV). A total of 120 elderly patients with lung cancer were included in the present study. According to the random number table method, these patients were randomly divided into two groups: group D and group C. Patients in group D were intravenously pumped with 0.5 μg/kg/h of DEX before anesthesia. The infusion was completed within 15 min, and anesthesia was induced by venous injection. Patients in group C were pumped with equal volumes of normal saline. At T2 and T3, compared with group C, group D had a significant decrease in cardiac index, mean arterial pressure and central venous pressure (p < 0.05). At T2, T3 and T4, compared with group C, group D had a significant increase in pH and PaO2 (p < 0.05). At T2, T3 and T4, compared with group C, group D had a significant decrease in Qs/Qt (p < 0.05). At T6, compared with group C, group D had a significant decrease in the supernatant of bronchoalveolar lavage fluid of tumor necrosis factor-α and interleukin 6 (p < 0.05). At T5, compared with group C, group D had a significant decrease in Visual Analogue Scale score (p < 0.05), and a significant increase in Ramsay Sedation Scale score (p < 0.05), and the number of respiratory and cardiovascular events also decreased (p < 0.05). In elderly patients, dexmedetomidine can reduce Qs/Qt and increase PaO2 during OLV in surgery. It can reduce lung injury. Moreover, DEX reduced respiratory and cardiovascular complications in the perioperative period.

Geranylgeranyl diphosphate synthase 1 knockout ameliorates ventilator-induced lung injury via regulation of TLR2/4-AP-1 signaling

ObjectiveTo investigate the role of geranylgeranyl diphosphate synthase 1 (GGPPS1) in ventilator-induced lung injury along with the underlying mechanism. MethodsA murine VILI model was induced by high-tidal volume ventilation in both wild-type and GGPPS1 knockout mice. GGPPS1 expression was detected in the bronchoalveolar lavage fluid (BALF) supernatants of acute respiratory distress syndrome (ARDS) patients and healthy volunteers, as well as in lung tissues and BALF supernatants of the VILI mice using enzyme-linked immunosorbent assay (ELISA), quantitative reverse transcription polymerase chain reaction (qRT-PCR), western bolt and immunohistochemical (IHC). The wet/dry ratio, total BALF proteins, and lung injury score were analyzed. The percentage of neutrophils was detected by flow cytometry and IHC. Inflammatory cytokine levels were measured by ELISA and qRT-PCR. The related expression of Toll-like receptor (TLR)2/4 and its downstream proteins was evaluated by western blot. ResultsGGPPS1 in BALF supernatants was upregulated in ARDS patients and the VILI mice. Depletion of GGPPS1 significantly alleviated the severity of ventilator induced lung injury in mice. Total cell count, neutrophils and inflammatory cytokines (interleukin [IL]-6, IL-1β, IL-18 and tumor necrosis factor-α) levels in BALF were reduced after GGPPS1 depletion. Moreover, addition of exogenous GGPP in GGPPS-deficient mice significantly exacerbated the severity of ventilator induced lung injury as compared to the PBS treated controls. Mechanistically, the expression of TLR2/4, as well as downstream proteins including activator protein-1 (AP-1) was suppressed in lung tissues of GGPPS1-deficient mice. ConclusionGGPPS1 promoted the pathogenesis of VILI by modulating the TLR2/4–AP-1 signaling pathway, and GGPPS1 knockout significantly alleviated the lung injury and inflammation in the VILI mice.