Search Results (123)

Background: The objective was to identify management strategies of IFI in critically ill patients through a Spanish national survey. Methods: A cross-sectional multicentre survey among ICU specialists, experienced in IFI, was performed (22 April–25 July 2024). The survey consisted of 13 questions with four closed answers. Results: Sixty-three specialists from 51 hospitals of 16 regions completed the survey. 95% stated that, in high-risk patients with clinical suspicion of Pulmonary Aspergillosis (PA), galactomannan in BAL is performed to guide treatment. In the treatment of patients with PA and influenza, 86% declared that isavuconazole and liposomal amphotericin B are recommended treatments and in high suspicion of Aspergillus coinfection, 76% recommended empirical treatment waiting for microbiological confirmation. 90% declared that the use of Extracorporeal Membrane Oxygenation (ECMO) and Renal Replacement Therapies (RRT) could be associated with lower azole levels. Regarding intra-abdominal candidiasis, 78% that physiopathological changes in critically ill patients, reduce their entry into peritoneal fluid. Conclusions: The majority of the respondents agreed (>80%) on: In suspicion of PA, Galactomannan in BAL to guide treatment is mandatory; In case of aspergillosis and influenza, isavuconazole and liposomal amphotericin B are the recommended treatments; The use of ECMO and RRT could be associated with lower azole levels. Full article

The secondary immunomodulatory effects of conventional therapeutics, such as antibiotics and cytostatics, are frequently overlooked despite their significant clinical implications. Building on our previous findings that drugs like paclitaxel and doxorubicin heavily influence macrophage polarization—potentially driving metastasis or inflammation—this study systematically evaluates the secondary immune-modulating actions of standard drugs and natural adjuvants. Using patient-derived bronchoalveolar lavage (BAL) fluid (ex vivo alveolar macrophages), we developed an analytical platform using synthetic carbohydrate-functionalized fluorescent ligands targeting key receptors (CD206, CD209, CD280, CD301). Integrating ligand-binding profiles with Linear Discriminant Analysis (LDA) yielded quantitative immune-state vectors capable of differentiating favorable and unfavorable prognostic signatures and imbalanced immune states. Pro-filing samples across heterogeneous respiratory conditions revealed highly con-text-dependent responses. While some treatments synergistically corrected unfavorable imbalanced profiles, others provoked dysregulation. Notably, in pneumonia or bronchitis with an asthma-prone M2-dominant profile, specific antibiotic regimens are critical; doxycycline, for instance, may exacerbate patient deterioration by further driving M2a polarization. Crucially, we identified that natural adjuvants (e.g., curcumin, coumarins, polyphenols) exhibit potent properties capable of correcting these adverse secondary drug effects. Ultimately, this profiling platform highlights the necessity of evaluating patient-specific secondary drug effects, offering a functional blueprint for precision immunotherapy and the rational design of adjuvant-enhanced treatments. Full article

Asthma, chronic obstructive pulmonary disease (COPD), and acute respiratory distress syndrome (ARDS) are the major lung inflammatory complications affecting the global population. Exposure to allergens, infections, smoking, and environmental pollutants could cause persistent oxidative stress and dysregulated immune responses, leading to lung inflammatory complications. Increased oxidative stress can lead to lipid peroxidation and the formation of toxic lipid aldehydes. One of the major lipid aldehydes formed during lipid peroxidation is 4-hydroxy-2-nonenal (4-HNE). 4-HNE is well known to covalently modify proteins, nucleic acids, and lipids, thus modifying cellular signaling pathways and inflammatory cascades. Increased levels of 4-HNE have been identified in lung tissues, bronchoalveolar lavage (BAL) fluid, and the serum of patients with inflammatory lung conditions. Further, 4-HNE contributes to airway remodeling, mitochondrial dysfunction, and modulation of inflammatory responses in the lung epithelial cells. Recent studies also indicate the potential role of 4-HNE as an important mediator and a potential biomarker of various human disease progression, including the diagnosis and monitoring of lung inflammatory diseases. In this narrative review, we discuss current evidence on the pathological role of 4-HNE, its potential as a biomarker, and its importance for early detection and for potential therapeutic strategies in lung inflammatory complications. Full article

Background: Idiopathic pulmonary fibrosis (IPF) is a progressive interstitial lung disease with a poor prognosis. Esophageal dilation and hiatal hernia are common in IPF and may facilitate microaspiration, exacerbating inflammation. We investigated the relationship between radiological esophageal dilation, smoking status, and bronchoalveolar lavage (BAL) cellular and immunological profiles in IPF patients. Methods: This retrospective study included 71 IPF patients. Esophageal diameters were measured at four levels (L1–L4) via high-resolution computed tomography (HRCT). BAL fluid was analyzed for differential cell counts, the Lipid-Laden Macrophage Index (LLMI), and T-lymphocyte subsets (CD4+, CD8+) using flow cytometry. Results: Esophageal dilation (diameter >10 mm) was present in 52.1% of patients, and 36.6% had a hiatal hernia. A significant negative correlation was found between distal esophageal dilation (L4) and BAL CD4+ counts (r = −0.267, p = 0.024). Similarly, the mean maximum esophageal diameter negatively correlated with BAL CD4+ levels (r = −0.288, p = 0.015). Patients with hiatal hernia had significantly higher BAL neutrophil percentages than those without (20.0% ± 4.39% vs. 8.93% ± 2.0%, p = 0.047). Furthermore, smokers exhibited significantly lower BAL CD4+ levels than non-smokers (p = 0.042). No significant correlation was found between esophageal dilation and the LLMI (p > 0.05). Conclusions: Esophageal dilation is significantly associated with altered local immune profiles in IPF. The negative correlation between distal esophageal dilation and BAL CD4+ counts, plus the link between hiatal hernia and neutrophilic inflammation, suggests an interplay between esophageal dysfunction and the pulmonary immune microenvironment. Radiological assessment of esophageal dilation may serve as a non-invasive surrogate marker for identifying high-risk clinical phenotypes in IPF. Full article

Objective: This study aimed to investigate the relationship between bronchoalveolar lavage (BAL) cellular profiles, microbiological status, and clinical outcomes such as hospital admission in adult patients with non-cystic fibrosis bronchiectasis. Methods: A retrospective cross-sectional study was conducted on thirty adult bronchiectasis patients. Demographic, clinical, and laboratory data were collected. The cellular components of BAL fluid (macrophages, neutrophils, lymphocytes, and eosinophils) were analyzed. Patients were grouped according to the presence of microbial culture growth and history of hospitalization in the past year. Statistical analyses were performed to determine significant relationships. Results: The median age was 57 years, and the gender distribution was equal. There was no significant difference in BAL cellular profiles between groups with and without culture growth. However, in the group with a hospital admission in the past year, BAL showed a significantly lower percentage of alveolar macrophages (20% vs. 47%, p = 0.011) and a higher percentage of eosinophils (5% vs. 1%, p = 0.036). The hospitalized group also showed a trend toward a higher neutrophil percentage and a lower lymphocyte/neutrophil ratio. Furthermore, surprising associations were noted, such as a higher BAL macrophage count in married individuals and higher BAL eosinophilia in patients with diabetes. Conclusions: BAL cellular analysis provides valuable information beyond routine microbiological investigations in bronchiectasis. The low-alveolar-macrophage and high-eosinophil profile was found to be significantly associated with hospitalization, and this profile has the potential to serve as a prognostic biomarker in defining the “high-risk” phenotype. These findings highlight the complexity of the local inflammatory response and reveal the potential role of BAL in developing personalized treatment strategies for patients with bronchiectasis. Full article

Background: Pleurodesis is a treatment method that aims to create permanent adhesion between the pleural layers to prevent recurrent fluid or air accumulation in the pleural cavity. Talc, one of the most commonly preferred agents in this procedure, is widely used in clinical practice. In this study, a new talc formulation with a modified surface to impart antibacterial and analgesic properties was experimentally evaluated for the first time. The main objective of the study was to comparatively assess the inflammatory and fibrotic responses following standard talc and modified talc applications. Methods: Thirty-six 12-week-old female Wistar albino rats were simply randomly divided into three different groups: control (n = 12), standard talc (n = 12), and modified talc (n = 12). Under anesthesia, 1 mL of physiological saline containing 17 mg of talc was injected intrapleurally into the right hemithorax. The presence of pneumothorax after the procedure was assessed by chest radiography. After a 12-day follow-up period, the animals were euthanized. Bronchoalveolar lavage (BAL) fluid samples, blood samples, and lung and pleural tissue samples were collected for biochemical, histopathological, and immunohistochemical analyses. Results: Modified talc application resulted in a significant increase in both visceral and parietal pleural thickness (p < 0.05). Granulation tissue formation and collagen deposition were significantly higher in the modified talc group. In addition, TGF-β expression and CD68-positive macrophage count increased significantly in the modified talc group (p < 0.05). Inflammatory changes in the lung parenchyma were limited and not statistically significant. Conclusions: The modified talc formulation enriched with lidocaine and antibacterial agents produced a stronger inflammatory and fibrotic response compared to standard talc. These findings indicate that modified talc may increase the effectiveness of pleurodesis. Furthermore, the absence of significant lung parenchymal damage suggests that this treatment is locally effective and feasible. However, further long-term and advanced studies are needed to translate these results into clinical use. Full article

Intravesical Bacillus Calmette–Guérin (iBCG) immunotherapy is the standard adjuvant treatment of non-muscle-invasive bladder cancer (NMIBC). Among the potential complications, cases of mycotic aneurysms and acute respiratory distress syndrome (ARDS) are rare but can be life-threatening. Because prior reports have not included whole-genome sequencing (WGS) of clinical Mycobacterium bovis BCG (M. bovis BCG) isolates to assess whether the infecting strain acquires mutations in vivo, we performed WGS in a severe disseminated iBCG-related infection. A 72-year-old man with bladder cancer underwent iBCG instillation. Twelve months after the final instillation, the patient developed an abdominal aortic aneurysm, which was detected and treated with endovascular aneurysm repair (EVAR). Two months later, the patient presented with fever, abdominal pain, and septic shock. Contrast-enhanced computed tomography (CT) and ¹⁸F-fluorodeoxyglucose positron emission tomography/CT (FDG-PET/CT) showed rapid aneurysm enlargement. Ziehl–Neelsen staining and PCR of aortic material identified M. bovis BCG. Direct PCR on BAL fluid and urine was negative; however, BAL and urine culture subsequently grew M. bovis BCG, and PCR performed on the culture isolate confirmed M. bovis BCG. Despite combined antituberculosis triplet therapy (isoniazid, rifampicin, and ethambutol), the patient developed ARDS, which gradually improved after surgical management. WGS (with >96% genome coverage) showed the isolate was highly concordant with the vaccine strain and lacked additional virulence-associated mutations, including in esxM. This case illustrates that severe systemic iBCG-related complications can occur without detectable in vivo acquisition of virulence-enhancing mutations; however, interpretation is limited by the single-case design and the absence of host genetic susceptibility testing. Our findings underscore the need for prolonged vigilance regarding late-onset vascular and pulmonary complications after iBCG, and highlight the importance of early multidisciplinary management. Full article

Severe equine asthma (sEA) is characterized by increased lower airway neutrophils that contribute to dysregulated inflammation through the release of cytokines, reactive oxygen species and neutrophil extracellular traps (NETs). NETs are composed of cell-free DNA (cfDNA) intercalated with enzymatic proteins and are known to be increased in the lower airway of asthmatic horses. The objectives of this study were two-fold: 1. Determine if cfDNA can be accurately measured in equine bronchoalveolar lavage fluid (BALF) and tracheal wash (TW) with a Qubit 4 fluorometer. 2. Determine whether Qubit-measured cfDNA in BALF or TW is significantly different in horses with sEA, mild/moderate neutrophilic equine asthma, mastocytic equine asthma and healthy horses. A total of sixty-three horses received a physical examination and clinical score followed by a BAL +/− TW. Cell-free DNA was measured using three methods in unfiltered BAL and TW as well as BAL and TW supernatant. Cell-free DNA concentrations were highly correlated between the Qubit 4 fluorometer and NanoDrop spectrophotometer as well as between the Qubit 4 fluorometer and SYTOX green plate-based assay. Cell-free DNA concentrations were highly correlated between unfiltered TW and TW supernatant as well as between unfiltered BALF and BAL supernatant. Cell-free DNA concentrations in BAL and TW supernatant were significantly higher in horses with sEA compared to healthy horses or horses with mild/moderate equine asthma. Cell-free DNA is a biomarker of sEA that can be easily measured in the field with the small portable Qubit 4 fluorometer in BAL and TW fluid. These findings support further investigation of NETs as a biomarker and potential therapeutic target for severe equine asthma. Full article

We have developed a method for in vivo quantitation of lung delivery of inhaled nebulized drugs by measuring a fluorescent-labeled analog in bronchioalveolar lavage fluid (BALF) collected immediately after inhalation dosing. The effectiveness of delivery of an aerosolized formulation of our proprietary water-miscible vitamin A product to the deep lung (target organ) was studied; the product is being developed for prevention of bronchopulmonary dysplasia (BPD) in preterm infants. The fluorescent retinol analog was incorporated by spiking into a standard formulation, remaining fully compatible with existing nebulizer administration procedures for animal exposure. The method provides quantitation of the delivered dose (DD) to the lung within a few minutes after dosing; fluorescence in BAL in a plate reader allows for simple rapid quantitation of the delivered drug, while avoiding the complexities of other labeling methods (e.g., heavy labels or radioactivity). Data from newborn rat and lamb models showed linear dose responses, validating the method. Approximately 5–10% of the inhaled drug was recovered in BALF in both models, consistent with reports in the literature. The ease of use of the method facilitated various aspects of our project, including the transition to more clinically relevant animal models and aerosol exposure systems. The formulation of this approach could be spiked into other formulations, allowing application of the method to other aerosol drug development programs. Full article

Background/Objectives: Obesity and hyperglycemia predispose patients to respiratory infections. Although the lung is a major organ to utilize glucose, pulmonary glucose homeostasis in type 2 diabetic (T2Dx) subjects remains poorly characterized. We hypothesized that pulmonary glucose transport would be altered during T2Dx, which would be rescued with long-term metformin treatment. Methods: T2Dx was induced by feeding mice a high-fat diet for 16 weeks, with metformin treatment administered during the final 8 weeks. Results: Glucose transporter (GLUT) protein expression and trafficking was quantified by Western blotting and the biotinylated photolabeling assay, respectively. T2Dx mice exhibited obesity, and increased glucose levels in blood and bronchoalveolar lavage (BAL) fluid. T2Dx also significantly decreased protein expression of GLUTs from Class I (i.e., GLUT-2 and -4) and class III (i.e., GLUT-10 and -12) isoforms in lung. Metformin treatment restored the protein expression of GLUT-2, -4, and -10, but not GLUT-12. Pulmonary cell surface expression of GLUT-4 and -8 was also significantly reduced in T2Dx mice and rescued by metformin. Conclusions: These findings suggest that alterations in pulmonary GLUT expression and trafficking during diabetes could contribute to the elevated airway glucose levels and severity of respiratory infections. Metformin treatment restored pulmonary glucose transport during T2Dx. Full article

Mild–moderate equine asthma (MEA) is a very common but underdiagnosed pulmonary disease in horses, with mild cases not showing clinical respiratory signs. This study evaluates the influence of a standardized lunging exercise test (SLET) on bronchoalveolar lavage fluid (BALF) cytology in MEA horses. We hypothesized that SLET would increase the total nucleated cell count (TNCC) and/or percentages of inflammatory cells associated with EA. In a prospective, randomized, non-blinded, between-subjects study design of two independent groups, 39 horses (17 mild and 22 moderate) were included. They were chosen out of a cohort of horses undergoing respiratory investigations (history, clinical examination, and clinical pathology (white blood cells (WBC) and arterial blood gas analysis (aBGA)) consistent with MEA, using a scoring system in a clinical setting of an equine referral clinic. Bronchoalveolar lavage (BAL) was performed 30 min post-SLET in 16 randomly chosen horses. The other horses underwent BAL without SLET. The SLET resulted in a statistically significant increase (p < 0.001) in the proportions of neutrophils in BALF cytology, and in an increased chance of confirmation of the presumed diagnosis in horses with mild phenotypes (p < 0.001, OR = 8.00, CI = 1.28–50.04), while moderate phenotypes were less frequently diagnosed. Exercise had no association with cytology across all horses. Unexpectedly, the SLET group of horses with a moderate phenotype had a statistically significant lower TNCC (p = 0.035). In conclusion, an SLET prior to BAL might increase the chance of an MEA diagnosis. Full article

Conventional diagnostic methods (CDMs) for lower respiratory infections (LRIs) have limitations in detecting causative pathogens. This study evaluates the utility of shotgun metagenomic sequencing (SMS) as a complementary diagnostic tool using bronchoalveolar lavage (BAL) fluid. Sixteen BAL fluid samples from pneumonia patients with positive CDM results—including bacterial/fungal cultures; PCR for Mycobacterium tuberculosis or cytomegalovirus; and the BioFire^® FilmArray^® Pneumonia Panel (BioFire Diagnostics LLC, Salt Lake City, UT, USA)—underwent 10 Gb SMS on the Illumina NovaSeq 6000 platform (Illumina, San Diego, CA, USA). Reads were aligned to the NCBI RefSeq database; with fungal identification further supported by internal transcribed spacer (ITS) analysis. Antibiotic resistance genes (ARGs) were annotated using the Comprehensive Antibiotic Resistance Database. Microbial reads accounted for 0.00002–0.04971% per sample. SMS detected corresponding bacteria in 63% of cases, increasing to 69% when subdominant taxa were included. Fungal reads were low; however, Candida species were identified in four samples via ITS. No viral reads were detected. ARGs meeting perfect match criteria were found in two cases. This is the first real-world study comparing SMS with CDMs, including semiquantitative PCR, in BAL fluid for LRI. SMS shows promise as a supplementary diagnostic method, with further research needed to optimize its performance and cost-effectiveness. Full article

Background: Bronchoalveolar lavage fluid (BALF) analysis is essential for the accurate diagnosis and management of interstitial lung diseases (ILDs). Despite established guidelines, variability in sample handling may affect diagnostic accuracy. This study aimed to evaluate how different storage conditions impact BALF cell counts and differentials to guide optimal sample handling practices. Methods: Forty patients who underwent BAL at Chiba University Hospital from June to December 2024 were included. BALF samples were allocated into five groups based on processing conditions: immediate analysis within 1 h, storage at either at 4 °C or room temperature (RT) for 6 h, or storage at 4 °C or RT for 24 h. Total cell counts (TCC) and differential counts were measured and compared among conditions. Results: TCC remained stable over 24 h at both 4 °C (p = 0.86) and RT (p = 0.90). Similarly, the percentages of eosinophils, lymphocytes, and macrophages did not significantly change at either temperature (all p > 0.05). Notably, neutrophil percentages showed a significant decline over time under both storage conditions—at 4 °C (p = 0.02) and at room temperature (p < 0.01). Post hoc tests revealed a notable decreasing trend at 6 h and significant reductions by 24 h at 4 °C (p = 0.09 and p = 0.02, respectively), and significant decreases at both 6 and 24 h at RT (p = 0.01, <0.01). Conclusions: Among the various cell types in BALF, neutrophil proportions are particularly susceptible to storage conditions, showing a significant decline over time—especially at room temperature—while other cell types remain stable for up to 24 h. Therefore, prompt processing or appropriate refrigeration of BALF is essential to ensure reliable cytological analysis and accurate clinical interpretation. Full article

Background: Invasive aspergillosis (IA) is a life-threatening fungal infection that primarily affects immunocompromised individuals and has high morbidity and mortality rates, necessitating timely diagnosis and treatment. This study aimed to evaluate the prognostic utility of serum and bronchoalveolar lavage (BAL) fluid galactomannan levels, as well as galactomannan kinetics, in patients with IA. Methods: We retrospectively reviewed the medical records of patients who were diagnosed with proven or probable IA from March 2016 to April 2024 at a tertiary cancer center. The collected data included patient characteristics, baseline and peak galactomannan levels in serum and BAL fluid, galactomannan trends, and clinical outcomes. Subgroup analyses were performed to assess the prognostic value of dual-source galactomannan positivity (positive serum and BAL fluid galactomannan levels). Results: Elevated baseline serum galactomannan levels independently predicted treatment non-response (p = 0.039) and 12-week all-cause mortality (p < 0.001). Peak serum and BAL fluid galactomannan levels were strongly associated with poor clinical outcomes (p < 0.01). Compared to single-source galactomannan positivity, dual-source galactomannan positivity was linked to reduced treatment response (22% vs. 43%, p = 0.01) and higher IA-attributable mortality (52% vs. 27%, p = 0.002). Patients with neutropenia had poorer outcomes compared to patients without neutropenia, but neutrophil recovery dramatically improved survival (25% vs. 69% mortality, p < 0.0001). Early galactomannan kinetics and malignancy type had limited prognostic value. Conclusions: Our findings highlight the potential role of galactomannan as a key biomarker for early prognostication for IA. The strong association between galactomannan levels and clinical outcomes suggests its utility in identifying high-risk patients who may benefit from more aggressive management. Further studies are needed to introduce a nuanced and context-specific use of galactomannan into clinical practice and assess its role as a prognostic biomarker. Full article

Objective: To investigate the effect of Lipopolysaccharide (LPS)-induced acute lung injury (ALI) on the pulmonary pharmacokinetics (PK) of a systemically administered antibody in mice. Method: The PK of a non-target-binding antibody was evaluated in healthy mice and mice with intratracheal instillation of 5 mg/kg LPS. The plasma, bronchoalveolar lavage (BAL), trachea, bronchi, and lung homogenate PK of the antibody were measured following intravenous administration of 5 mg/kg antibody dose. Noncompartmental analysis was performed to determine AUC values. Antibody concentrations in all biological matrices were quantified using qualified ELISA. The effect of ALI on BAL albumin and total protein concentrations was also determined. BAL protein concentrations were corrected for dilution using plasma urea concentrations. Results: Intratracheal instillation of LPS and the resultant ALI led to ~2–4-fold higher concentrations of albumin and proteins in the BAL. LPS-induced ALI also notably altered the pulmonary PK of the antibody. The effect of ALI on the antibody PK was time and tissue dependent. The trachea and bronchi showed ~1.7-fold and ~1.4-fold lower antibody exposure compared with the control group, but the BAL fluid exhibited ~4-fold increase in antibody exposure following LPS treatment. Most noticeable changes in antibody PK occurred 24 h after LPS administration, and the effect was temporary for the bronchi and trachea. However, the changes in lung homogenate and, more notably, in BAL persisted until the end of the experiment. Thus, our investigation suggests that due to the acute nature of ALI-induced pathophysiology and the changing severity of the disease, the dose and timing of antibody administration following ALI may need to be optimized based on the target site of action (e.g., bronchi, trachea, BAL, lung parenchyma, etc.) to maximize the therapeutic effect of the antibody. Conclusions: ALI may significantly affect pulmonary PK of systemically administered antibodies. Changes caused by ALI are time and tissue dependent, and hence, the timing and dose of antibody following ALI may need to be optimized to maximize the therapeutic effect of the antibody at the site of action. Full article