Citations

Background: Cell-penetrating peptides cross cell membrane barriers while carrying cargoes in a functional form. Our work identified two novel lung-targeting peptides, S7A and R11A. Here, we present studies on biodistribution, the cell types targeted, and an in vitro proof of application. Methods: Studies were performed in human bronchial epithelial cells (HBECs) with and without various endocytic inhibitors, and coincubation with fluorescently labeled transferrin or endocytic markers. Cyclic R11A (cR11A) was conjugated to siRNA duplexes and anti-viral activity against SARS-CoV-2 was tested. Biodistribution studies were performed by injecting wild-type mice with fluorescently labeled peptides, and various circulation times were allowed for, as well as cross-staining of lung sections or isolated single cells with various cellular markers, followed by fluorescence-activated cell sorting or confocal microscopy. Results: cR11A showed peak uptake in 15 min, with the highest uptake in airway epithelial type II (ATII) cells, followed by p63+ basal cells and ionocytes. Cyclization increased transduction efficiencies ~100-fold. Endocytosis studies showed a decrease in peptide uptake by pre-treatment with Pitstop2 but not Amiloride or Nystatin. Endocytic marker Lamp1 showed colocalization at the earliest time point, with the escape of the peptide from endocytic vesicles later. cR11A conjugated to ant-spike and anti-envelop proteins showed anti-viral effects with an EC90 of 0.6 μM and 1.0 µM, respectively. Conclusions: We have identified a novel peptide, cR11A, that targets ATII, basal cells, and ionocytes, the cyclization of which increased transduction efficiency in vitro and in vivo. The uptake mechanism appears to be via clathrin-mediated endocytosis with escape from endocytic vesicles. cR11A can act as a vector to deliver anti-viral siRNA to epithelial cells.

Tandem exposure to cannabinoids and nicotine during pregnancy poses significant risks to fetal development and health including low birth weight, preterm birth, and perinatal death. However, the effect of co-exposure with the most prominent cannabinoids found in Cannabis sativa (Trans-Δ-9- tetrahydrocannabinol (THC) and cannabidiol (CBD)) alongside nicotine on fetal lung development is not well understood. This study aims to investigate the combinatorial impact of CBD, THC, and nicotine, on human lung development through investigating cellular senescence and the potential role of senolytics in alleviating these effects. Human fetal lung explants from 10 to 16 weeks gestation were cultured on air-liquid interface and treated with various concentrations of nicotine (1µM), CBD, and THC (5µM, or 2.5µM each when used in combination), for 72 hours. To assess the effect of senolytics, the explants were also treated for 48 hours with a combination of CBD, THC, and nicotine. Subsequently, senolytics, Dasatinib (D, 250nM) and Quercetin (Q, 375nM), were added or not for an additional 24 hours. After the 72-hour treatment, explants were collected and either fixed for immunofluorescence staining (IF) analyses or used for RNA extraction and quantitative reverse transcription PCR (qRT-PCR). Additionally, the supernatants were used to evaluate the expression of the senescence-associated secretory phenotype (SASP), conducted by Eve Technologies Corporation-Canada. This study used Luminex xMAP technology for multiplexed quantification of 96 Human cytokines, chemokines, and growth factors. Our results demonstrated that co-exposure to CBD, THC, and nicotine increases the expression of the senescence markers CDKN1A (0.1163±0.02384, p=0.0094, n=8) and CDKN2A (0.005793±0.002157, p=0.0516, n=8) encoding P21 and P16 respectively. Moreover, the IF staining showed increased expression of the DNA damage marker γH2A.X following treatment with CBD+THC+nicotine (7.375±2.154% vs 2.245±0.8329%, p=0.0604, n=6). Furthermore, the lung explants treated with cannabinoids and nicotine released cytokines and chemokines involved in immune responses and inflammation among others IL-6, IL-8, M-CSF, MCP-3, MIP-1β, TNFα, MIP-3β, and TSLP (p<0.05). The addition of D+Q reduced SASP secretion and showed a decreasing trend in expression of the senescence genes CDKN1A and CDKN2A compared to the triple treatment. Taken together, our results showed that the combinatorial exposure to CBD, THC, and nicotine increases senescence, DNA damage, and the senescence-associated secretory phenotypes (SASP) secretion. The use of senolytics (D+Q) may prove to be a promising therapeutic option, alleviating several of the negative effects induced by the different substances. RO1HL171915-01A1 RO!HL155104 RO1HL158532-03 ROIHL160570 This abstract was presented at the American Physiology Summit 2025 and is only available in HTML format. There is no downloadable file or PDF version. The Physiology editorial board was not involved in the peer review process.

Abstract Aims Syncope remains a diagnostic challenge despite advancements in testing and treatment. Cardiac syncope is an independent predictor of mortality and can be difficult to distinguish from other causes of transient loss of consciousness (TLOC). This paper explores whether artificial intelligence (AI) can improve the evaluation and management of patients with syncope. Methods and results We conducted a literature review and incorporated the opinions of experts in the fields of syncope and AI. The cause of TLOC is often unclear, hospitalization criteria are ambiguous, diagnostic tests are frequently non-informative, and assessments are costly. Patients are left with unanswered questions and limited guidance. Artificial intelligence (AI) has the potential to optimize syncope evaluation by processing large data sets, detecting imperceptible patterns, and assisting clinicians. However, AI has limitations, including errors, lack of human empathy, and uncertain clinical utility. Liability issues further complicate its integration. We present three viewpoints: (i) AI is crucial for advancing syncope management; (ii) AI can enhance the patient experience; and (iii) AI in syncope care is inevitable. Conclusion Artificial intelligence may improve syncope diagnosis and management, particularly through machine learning–based test interpretation and wearable device data. However, it has yet to surpass human clinical judgment in complex decision-making. Current challenges include gaps in understanding syncope mechanisms, AI interpretability, generalizability, and clinical integration. Standardized diagnostic approaches, real-world validation, and curated data sets are essential for progress. Artificial intelligence may enhance efficiency and communication but raises concerns regarding confidentiality, bias, inequities, and legal implications.

AimsPolymyalgia rheumatica (PMR) is a chronic autoimmune disorder that mainly affects older adults. Pulmonary disease in PMR is rare but may be under‐recognized and pathological descriptions thereof are few. We aimed to characterize diffuse parenchymal lung disease (DPLD) in PMR.Methods and resultsInstitutional archives were searched for patients having PMR and DPLD with lung tissue sampling. After excluding cases with infection, concomitant rheumatoid arthritis, or smoking‐related DPLD only, 11 patients (9 women, median age 75 years) were enrolled. Clinical history and pathology slides were reviewed. One of the 11 patients (9%) had concomitant giant cell arteritis; the remaining patients had no other rheumatological diseases. All had been treated for PMR with immunosuppression, and most presented years later (median 6 years) with non‐specific respiratory symptoms. Radiographically, bilateral ground‐glass opacities and reticulation were typical and were usually lower lobe predominant. Histologically, fibrosis was seen in 8 of 11 (73%) patients and was unclassifiable in four; non‐specific interstitial pneumonia was encountered in three patients, and usual interstitial pneumonia was seen in only one case. Evidence of acute lung injury occurred in 9 (82%) patients, including three with acute lung injury only, usually manifesting as organizing pneumonia. Diffuse alveolar haemorrhage was seen in four cases (29%), including two patients with haemoptysis and capillaritis.ConclusionsOur data corroborate prior reports of clinically significant DPLD in some patients with PMR. Histopathological findings mirror other rheumatological disorders and include diffuse alveolar haemorrhage with capillaritis. Additional studies are warranted to clarify the association between PMR and DPLD.

Abstract Background β-Quantification (BQ) is the reference method for low-density lipoprotein cholesterol (LDL-C) determination. It is not widely available, making it challenging for laboratories to assess the accuracy of LDL-C methods. Our goal was to develop an indirect graphical approach for comparing LDL-C test results to the BQ reference method. Methods BQ results from Mayo Medical Laboratories (n = 39 969) and the National Institutes of Health (n = 17 825) were used to investigate the interrelationships between lipid panel tests. A plot of LDL-C/non high-density lipoprotein cholesterol (nonHDL-C) vs (triglyceride (TG)/nonHDL-C)0.5 (lipid ratio plot) resulted in a negative linear regression line (y = −34.2x + 115). Based on simulation analysis, the minimum sample size for calculating its slope and intercept with a coefficient of variation of about 7.5% was 80. Results The regression lines for the lipid ratio plot of LDL-C calculated by the Sampson–NIH (y = −37.6x + 118) and enhanced Sampson–NIH (y = −33.1x + 116) equations closely overlapped with the BQ method. In contrast, the Martin–Hopkins equation showed a positive bias with an increasing TG/nonHDL-C ratio and exceeded the recommended bias limit of 4% on hypertriglyceridemic samples (y = −25.5x + 107). The Friedewald equation showed an even larger negative bias with hypertriglyceridemia (y = −47.5x + 126). Lipid ratio plots of the Roche direct assay revealed a fixed positive bias of approximately 4% (y = −33.9x + 120), whereas a much larger proportional positive bias was observed with increasing triglycerides for the Beckman direct assay (y = −14.6x + 97.8). Conclusions The lipid ratio plot is a simple graphical approach that can be readily performed by clinical laboratories for investigating the accuracy of LDL-C assays by calculation methods or direct assays.

ABSTRACT Introduction Calcineurin inhibitors (CNIs) are the cornerstone of lung transplant immunosuppression but are associated with nephrotoxicity and other adverse effects. Although dose reduction and combination with Sirolimus, a mammalian target of rapamycin inhibitor (mTORi), have been explored, full conversion to Sirolimus remains uncommon. This study describes the characteristics and outcomes of lung transplant recipients who underwent complete CNI withdrawal and conversion to Sirolimus. Methods This retrospective case series included 52 lung transplant recipients transitioned to Sirolimus‐based immunosuppression between 2010 and 2021. Data on demographics, transplant characteristics, immunosuppression, and outcomes were collected from electronic medical records. Results The cohort included 31 males (59.6%) with a median age of 58.4 years (IQR 52.3–63.5). The primary indication for CNI withdrawal was renal dysfunction (73.1%). Sirolimus discontinuation occurred in 69.2%, most commonly due to edema (25.0%), lung toxicity (19.4%), rejection (19.4%), and surgical needs (22.2%). Clinically significant rejection occurred in 13.5%, with four patients progressing to chronic lung allograft dysfunction (CLAD) and two deaths. Sixteen patients (30.8%) tolerated CNI withdrawal for over a year. Conclusion Full conversion from CNI to Sirolimus is poorly tolerated due to side effects and has a high incidence of rejection. Further studies are needed to optimize patient selection and risk mitigation strategies.

INTRODUCTION Gastroesophageal reflux is common in respiratory disease, but the interplay between gastrointestinal mechanisms that expose individuals to reflux and potentially aspiration, and lung mechanics and function remain incompletely understood. Our aim was to investigate this in patients with chronic obstructive pulmonary disease (COPD) and non-idiopathic pulmonary fibrosis (IPF) interstitial lung disease (non-IPF ILD), and compare with our published findings in IPF. METHODS Fifty-seven patients with COPD (aged: 34–75 years) and 64 with non-IPF ILD (22–75 years) who underwent high-resolution impedance manometry and 24-hour pH impedance together with pulmonary function assessment were compared with 35 IPF patients (51–84 years). RESULTS COPD patients were less likely to exhibit ineffective esophageal motility (IEM) and/or absent contractility ( P = 0.009; P = 0.028), and tended to exhibit esophagogastric junction outflow obstruction (EGJOO) and/or hypercontractility ( P = 0.09, P = 0.14) than IPF and non-IPF ILD patients. Notably, integrated relaxation pressure correlated with esophageal length index (ELI) ( P = 0.048) and inspiratory LESP ( P = 0.003), with latter 2 correlating with each other ( P < 0.001). EGJOO patients tended to have fewer proximal reflux events and reduced pulmonary function, with the latter inversely correlating with ELI ( P < 0.05). Non-IPF ILD patients were less likely to exhibit EGJOO than COPD patients ( P = 0.27), and less likely to exhibit IEM ( P = 0.07) than IPF patients. However, those with IEM or EGJOO exhibited greater proportions of reflux events reaching the proximal esophagus than those with normal motility ( P < 0.03), which in contrast to IPF, seemed not to associate with worse pulmonary function. DISCUSSION Associations between esophageal motility, and lung mechanics and function, and consequently reflux, are very disease-specific.

Abstract Background Real-world first-line maintenance (1LM) treatment data are limited for advanced/metastatic non-small cell lung cancer (a/mNSCLC). Materials and Methods In this electronic health record-derived, deidentified database study, eligible patients (≥18 years; diagnosed with stage III/IV non-small cell lung cancer [June 1, 2017-September 30, 2021]) initiated pembrolizumab-based 1LM after 4-6 cycles of first-line (1L) platinum-based chemotherapy-pembrolizumab ± pemetrexed. Study outcomes were real-world time to next treatment or death (rwTTNTD), overall survival (rwOS), and predictors of outcomes. Results Of 1944 patients analyzed (median follow-up, 12.2 months), 51.9% received 1LM pembrolizumab-pemetrexed and 48.1% pembrolizumab monotherapy. Median rwTTNTD and rwOS were 9.2 (95% CI: 8.5-9.8) and 18.7 (95% CI: 17.7-20.3) months, respectively. In multivariable analyses, factors significantly associated with shorter rwTTNTD included 5%-&lt;10% (hazard ratio [HR], 1.34; 95% CI: 1.17-1.54) or ≥10% (HR, 1.69; 95% CI: 1.42-2.01) weight loss during 1L versus 0% or &lt;5% weight loss. Programmed death-ligand 1 (PD-L1) expression 1%–49% (HR, 0.81; 95% CI: 0.71-0.93) or ≥50% (HR, 0.55; 95% CI: 0.47-0.64) and female sex (HR, 0.85; 95% CI: 0.75-0.95) were significantly associated with longer rwTTNTD. These variables were also significantly associated with shorter (weight loss 5%-&lt;10%: HR, 1.52; 95% CI: 1.30-1.77; ≥10% HR, 2.06; 95% CI: 1.71-2.48) and longer rwOS (PD-L1 expression: 1%-49% HR, 0.84; 95% CI: 0.72-0.98; ≥ 50% HR, 0.57; 95% CI: 0.48-0.68; female sex: HR, 0.81; 95% CI: 0.71-0.92). Conclusions Predictors of real-world clinical outcomes included 1L treatment, weight loss, PD-L1 status, and sex. Poor outcomes persisted despite immunotherapy-based 1LM availability, revealing an unmet need in this population.

Abstract Background Cardiovascular guidelines have long recommended low-density lipoprotein-cholesterol (LDL-C) as the primary target for lipid-lowering therapy. Recent guidelines have emphasized the importance of achieving low LDL-C levels; hence, the accurate measurement of low LDL-C is increasingly clinically relevant. Methods Using lipid panel test results from the Mayo Clinic (n = 24 590) and the FOURIER clinical trial of evolocumab (n = 9605), the following modified Sampson equation was developed by least-squares regression to match LDL-C (mg/dL) by the β-quantification reference method, by combining terms into non High Density Lipoprotein Cholesterol (nonHDLC = Total Cholesterol – High Density Lipoprotein Cholesterol) and forcing the coefficient to be one: Results The modified Sampson equation demonstrated significant improvement in its concordance to the reference method compared to other equations (the Lin Concordance Correlation Coefficient 0.992, P &lt; 0.001). By overall kappa analysis, it showed the best agreement to the reference method at the 55 mg/dL cutpoint (1.4 mmol/L, 0.98 [P &lt; 0.001], Sampson–NIH: 0.96, Martin–Hopkins: 0.96, Friedewald: 0.94) and the 70 mg/dL cutpoint (1.8 mmol/L, 0.97 [P &lt; 0.001], Sampson–NIH: 0.94, Martin–Hopkins: 0.95, Friedewald: 0.92). The false classification rate of the modified Sampson equation was also significantly lower compared to the other equations at 55 mg/dL (15%, [P &lt; 0.001], Sampson–NIH: 29%, Martin–Hopkins: 28%, Friedewald: 37%) and 70 mg/dL (18%, [P &lt; 0.001]; Sampson–NIH: 30%, Martin–Hopkins: 28.%, Friedewald: 34%). The new equation increases the percentage of correctly classified patients with low LDL-C by approximately 10% to 20% over the other equations based on its net reclassification index. Conclusions The modified Sampson equation shows improved accuracy compared to other equations for low LDL-C. It more accurately identifies high-risk patients, who are not at their LDL-C goals and could benefit from more intensive lipid-lowering therapy.