Evolution of epithelial sodium channels: current concepts and hypotheses

The conquest of freshwater and terrestrial habitats was a key event during vertebrate evolution. Occupation of low-salinity and dry environments required significant osmoregulatory adaptations enabling stable ion and water homeostasis. Sodium is one of the most important ions within the extracellular liquid of vertebrates, and molecular machinery for urinary reabsorption of this electrolyte is critical for the maintenance of body osmoregulation. Key ion channels involved in the fine-tuning of sodium homeostasis in tetrapod vertebrates are epithelial sodium channels (ENaCs), which allow the selective influx of sodium ions across the apical membrane of epithelial cells lining the distal nephron or the colon. Furthermore, ENaC-mediated sodium absorption across tetrapod lung epithelia is crucial for the control of liquid volumes lining the pulmonary surfaces. ENaCs are vertebrate-specific members of the degenerin/ENaC family of cation channels; however, there is limited knowledge on the evolution of ENaC within this ion channel family. This review outlines current concepts and hypotheses on ENaC phylogeny and discusses the emergence of regulation-defining sequence motifs in the context of osmoregulatory adaptations during tetrapod terrestrialization. In light of the distinct regulation and expression of ENaC isoforms in tetrapod vertebrates, we discuss the potential significance of ENaC orthologs in osmoregulation of fishes as well as the putative fates of atypical channel isoforms in mammals. We hypothesize that ancestral proton-sensitive ENaC orthologs might have aided the osmoregulatory adaptation to freshwater environments whereas channel regulation by proteases evolved as a molecular adaptation to lung liquid homeostasis in terrestrial tetrapods.

Comprehensive Quantitative Assessment of Lung Liquid Clearance by Lung Ultrasound Score in Neonates with No Lung Disease during the First 24 Hours

Objectives. To comprehensively and quantitatively assess the process of lung liquid clearance using the lung ultrasound score. This study is to evaluate the whole healthy lungs of neonates during the first 24 h. Methods. Lung ultrasound was performed in neonates with no respiratory symptoms within 3 h after birth, and scans were then repeated at 6 hours and 24 hours, respectively. The entire chest wall was divided into 12 regions. The lung ultrasound scores of the anterior, posterior, upper, and lower regions and sum of all regions were calculated according to the ultrasound pattern of each region examined. Results. The total lung ultrasound score decreased gradually during the first 24 h, with the total lung ultrasound score at 6 h being significantly lower than that at <3 h (P<0.05). At <3 h, B-lines were more abundant in the posterior chest than in the anterior chest (P<0.001), and more B-lines were observed in the lower chest than in the upper chest (P<0.001). At 6 h and 24 h, there were no significant differences among the regions. Conclusion. Changes in the lung ultrasound score may quantitatively reflect the characteristics of different regions and processes of lung liquid clearance during the first 24 h.

Complete Lung Ultrasound Using Liquid Filling: A Review of Methods Regarding Sonographic Findings and Clinical Relevance

(200w) Lung ultrasound (LUS) is widely used for the diagnosis of pulmonary diseases such as solid nodules and consolidations in contact with the pleural cavity. However, sonography for processes of central disease remains impaired due to total sound reflection at the air tissue interfaces in the ventilated lung. These acoustic barriers can be overcome by replacing intra-alveolar air with liquid. Such filling has been reported using perfluorocarbon, saline or emulsions out of those. In order to achieve acoustic access enabling the use of LUS, complete gas free content is required. Such lung tissue - liquid compound will have untypical physical properties that might impact upon the sonographic visualization of central structures. Up to now, the filling of the lung has been reported for very specific applications and not classified regarding their sonographic findings. This work was therefore motivated to review the literature for methods of lung liquid instillation, classifying their methodological strength and limitations for achieving acoustic access and sonographic findings. Finally, their use for ultrasound based clinical applications will be discussed and the need for research will be outlined.

Delivery of Positive End-Expiratory Pressure Using Self-Inflating Bags during Newborn Resuscitation Is Possible Despite Mask Leak

<b><i>Background:</i></b> Ventilation is the key intervention to resuscitate non-breathing newborns. Positive end-expiratory pressure (PEEP) may facilitate lung-liquid clearance and help establish functional residual capacity. <b><i>Objectives:</i></b> The aim of this study was to describe how mask leak and ventilation rates affect delivered PEEP and tidal volumes during newborn resuscitations using a self-inflating bag with an integrated PEEP valve. <b><i>Methods:</i></b> This was an observational study including near-term/term newborns who received bag-mask ventilation (BMV) with a new self-inflating bag with a novel 6 mbar PEEP valve, without external gas flow, between October 1, 2016 and June 30, 2018 in rural Tanzania. Helping Babies Breathe-trained midwives performed most of the resuscitations. Pressures and flow were continuously measured and recorded by resuscitation monitors. <b><i>Results:</i></b> In total, 198 newborns with a median gestation of 39 weeks (25th, 75th percentiles 37, 40) and birth weight of 3,100 g (2,580, 3,500) were included. The median delivered PEEP and expired (tidal) volume at different levels of mask leak were 6.0 mbar and 11.3 mL/kg at 0–20% mask leak, 5.5 mbar and 9.3 mL/kg at 20–40%, 5.2 mbar and 7.8 mL/kg at 40–60%, 4.6 mbar and 5.0 mL/kg at 60–80%, and 1.0 mbar and 0.6 mL/kg at 80–100% mask leak. A high ventilation rate (&#x3e;60/min) nearly halved expired volumes compared to &#x3c;60/min for 0–60% leak. The BMV rate had a negligible effect on peak inflation pressure (PIP) and PEEP. <b><i>Conclusions:</i></b> Mask leak up to 80% did not impair the provision of recommended PEEP or tidal volumes during BMV with a self-inflating bag. High or low ventilation rates did not significantly affect PIP or PEEP. Expired volumes were reduced at ventilation rates &#x3e;60/min.

Tracking plasma KRAS mutations (mu) in lung adenocarcinoma (LUAC) patients (p) and branching evolution.

9055 Background: KRAS m in LUAC p are recalcitrant to therapy. In mice models and p, STK11 m confer poor prognosis. Patients with KRAS, or KRAS with TP53 m, benefit from immunotherapy (IO). We used a cell free DNA (cfDNA) sequencing platform to sub-group 53 LUAC p with KRAS m from the Spanish Lung Liquid versus Invasive biopsy Program (SLLIP, NCT03248089), according to the coexistence of TP53 and STK11 m. We evaluated the treatment outcome in the KRAS subgroups and we explored the mutational evolution at 2 weeks (w), and at the end of the study (EOS). Methods: SLLIP was a multi-center observational study in patients with treatment-naïve metastatic LUAC. Genotyping, with a clinically validated cfDNA assay (Guardant360) was performed at 3 time points: before start of treatment, at 2 w, and upon progression or at 12 months (mo). Oncoprints were constructed based on mutation status and variant allele frequency (VAF). Results: 53 p with KRAS alterations were included. 36 male; median age 59; 46 current/ex-smokers; 64% received 1st line platinum-based chemotherapy (CT), 4% platinum-based CT+IO, 8% platinum-based CT plus bevacizumab, and 24% other or no therapy. 13 p had only KRAS m (K-only group), 25 p had KRAS + TP53 m (KP group), and 15 p had KRAS + STK11 with or without TP53 m (KS group). Median progression-free survival was 3.5 mo for all 53 p, 4.8 mo for the K-only group, 4.4 mo for the KP group, and only 2.6 mo for the KS group (p = 0.05 for KS versus K-only). The average VAF for K-only, KP, and KS groups at EOS were 6.4%, 9.7%, and 46%, respectively. When looking at p with cfDNA analysis at the three time points, the following were observed: In the K-only group, 25% lost KRAS m at 2 w and 50% at EOS. 50% and 75% gained TP53 m, at 2 w and EOS, respectively. None gained STK11 at the 2 time points. In the KP group, 40% lost KRAS m at 2 w but all had KRAS m at EOS. 20% and 10% lost TP53 m, at 2 w and EOS, respectively. 10% gained STK11 m at the 2 time points. In the KS group, 33% lost KRAS m at the 2 time points. All and 33% lost STK11 m at 2 w and EOS, respectively. 66% gained TP53 at the 2 time points. Conclusions: Subgroups of KRAS m traced in cfDNA confirm dismal prognosis to 1st line therapies. These findings prompt us to tailor IO for K-only or KP subgroups. For p with STK11 m, restoration of the STK11 function is warranted. Clinical trial information: NCT03248089.

Reducing lung liquid volume increases biventricular outputs and systemic arterial blood flows despite decreased cardiac filling pressures in fetal lambs

As prior work has shown that reducing lung liquid volume 1) increases pulmonary arterial (PA) blood flow, 2) augments right ventricular (RV) output/power, and 3) decreases left atrial (LA) pressure, we tested the hypothesis that this perturbation has global cardiovascular effects. Ten anesthetized, open-chest fetal lambs (128 ± 2 days gestation, full term = 147 days) were acutely instrumented with 1) LA and right atrial (RA) catheters, 2) aortic and pulmonary trunk catheters, 3) brachiocephalic trunk, aortic isthmus, ductal, and left PA flow probes to obtain left ventricular (LV) and RV outputs and hydraulic power and flow in the descending thoracic aorta, and 4) an endotracheal tube to remove lung liquid. A 17 ± 7 ml/kg reduction of lung liquid volume 1) decreased LA and RA pressures similarly (1.5–1.6 mmHg, P < 0.001), 2) augmented LV and RV outputs (21–24%, P < 0.001) and total power (27–28%, P < 0.005), 3) increased systolic flows in the brachiocephalic trunk (18%, P < 0.001), aortic isthmus (29%, P < 0.005), ductus (12%, P < 0.005), and descending thoracic aorta (16%, P < 0.001), 4) increased mean PA flow via a higher systolic inflow (37%, P < 0.001) and lower diastolic backflow (−16%, P < 0.05), and 5) did not change systemic vascular conductance or arterial compliance but increased both pulmonary vascular conductance and arterial compliance (1.8-fold, P < 0.001). These data suggest that hemodynamic effects of lung liquid volume reduction are not confined to the lungs but extend to all cardiac chambers via rises in LV and RV outputs and power, despite falls in cardiac filling pressures, as well as the systemic circulation, via downstream increases in systolic flows of major central arteries.