Surface Area For Gas Exchange Research Articles

Dynamic Hippo pathway activity underlies mesenchymal differentiation during lung alveolar morphogenesis.

Alveologenesis, the final stage in lung development, substantially remodels the distal lung, expanding the alveolar surface area for efficient gas exchange. Secondary crest myofibroblasts (SCMF) exist transiently in the neonatal distal lung and are crucial for alveologenesis. However, the pathways that regulate SCMF function, proliferation and temporal identity remain poorly understood. To address this, we purified SCMFs from reporter mice, performed bulk RNA-seq and found dynamic changes in Hippo-signaling components during alveologenesis. We deleted the Hippo effectors Yap/Taz from Acta2-expressing cells at the onset of alveologenesis, causing a significant arrest in alveolar development. Using single cell RNA-seq, we identified a distinct cluster of cells in mutant lungs with altered expression of marker genes associated with proximal mesenchymal cell types, airway smooth muscle and alveolar duct myofibroblasts. In vitro studies confirmed that Yap/Taz regulates myofibroblast-associated gene signature and contractility. Together, our findings show that Yap/Taz is essential for maintaining functional myofibroblast identity during postnatal alveologenesis.

Dedifferentiated early postnatal lung myofibroblasts redifferentiate in adult disease.

Alveolarization ensures sufficient lung surface area for gas exchange, and during bulk alveolarization in mice (postnatal day [P] 4.5-14.5), alpha-smooth muscle actin (SMA)+ myofibroblasts accumulate, secrete elastin, and lay down alveolar septum. Herein, we delineate the dynamics of the lineage of early postnatal SMA+ myofibroblasts during and after bulk alveolarization and in response to lung injury. SMA+ lung myofibroblasts first appear at ∼ P2.5 and proliferate robustly. Lineage tracing shows that, at P14.5 and over the next few days, the vast majority of SMA+ myofibroblasts downregulate smooth muscle cell markers and undergo apoptosis. Of note, ∼8% of these dedifferentiated cells and another ∼1% of SMA+ myofibroblasts persist to adulthood. Single cell RNA sequencing analysis of the persistent SMA- cells and SMA+ myofibroblasts in the adult lung reveals distinct gene expression profiles. For instance, dedifferentiated SMA- cells exhibit higher levels of tissue remodeling genes. Most interestingly, these dedifferentiated early postnatal myofibroblasts re-express SMA upon exposure of the adult lung to hypoxia or the pro-fibrotic drug bleomycin. However, unlike during alveolarization, these cells that re-express SMA do not proliferate with hypoxia. In sum, dedifferentiated early postnatal myofibroblasts are a previously undescribed cell type in the adult lung and redifferentiate in response to injury.

Development of the terminal air spaces in the gray short-tailed opossum (Monodelphis domestica)- 3D reconstruction by microcomputed tomography.

Marsupials are born with structurally immature lungs when compared to eutherian mammals. The gray short-tailed opossum (Monodelphis domestica) is born at the late canalicular stage of lung development. Despite the high degree of immaturity, the lung is functioning as respiratory organ, however supported by the skin for gas exchange during the first postnatal days. Consequently, the majority of lung development takes place in ventilated functioning state during the postnatal period. Microcomputed tomography (μCT) was used to three-dimensionally reconstruct the terminal air spaces in order to reveal the timeline of lung morphogenesis. In addition, lung and air space volume as well as surface area were determined to assess the functional relevance of the structural changes in the developing lung. The development of the terminal air spaces was examined in 35 animals from embryonic day 13, during the postnatal period (neonate to 57 days) and in adults. At birth, the lung of Monodelphis domestica consists of few large terminal air spaces, which are poorly subdivided and open directly from short lobar bronchioles. During the first postnatal week the number of smaller terminal air spaces increases and numerous septal ridges indicate a process of subdivision, attaining the saccular stage by 7 postnatal days. The 3D reconstructions of the terminal air spaces demonstrated massive increases in air sac number and architectural complexity during the postnatal period. Between 28 and 35 postnatal days alveolarization started. Respiratory bronchioles, alveolar ducts and a typical acinus developed. The volume of the air spaces and the surface area for gas exchange increased markedly with alveolarization. The structural transformation from large terminal sacs to the final alveolar lung in the gray short-tailed opossum follows similar patterns as described in other marsupial and placental mammals. The processes involved in sacculation and alveolarization during lung development seem to be highly conservative within mammalian evolution.

Drebrin is Required for Myosin-facilitated Actin Cytoskeletal Remodeling during Pulmonary Alveolar Development.

Alveolar septation increases gas-exchange surface area and requires coordinated cytoskeletal rearrangement in lung fibroblasts (LFs) to balance the demands of contraction and cell migration. We hypothesized that DBN (drebrin), a modulator of the actin cytoskeleton in neuronal dendrites, regulates the remodeling of the LF cytoskeleton. Using mice bearing a transgelin-Cre-targeted deletion of Dbn in pulmonary fibroblasts and pericytes, we examined alterations in alveolar septal outgrowth, LF spreading and migration, and actomyosin function. The alveolar surface area and number of alveoli were reduced, whereas alveolar ducts were enlarged, in mice bearing the dbn deletion (DBNΔ) compared with their littermates bearing only one dbn-Flox allele (control). Cultured DBNΔ LFs were deficient in their responses to substrate rigidity and migrated more slowly. Drebrin was abundant in the actin cortex and lamella, and the actin fiber orientation was less uniform in lamella of DBNΔ LFs, which limited the development of traction forces and altered focal adhesion dynamics. Actin fiber orientation is regulated by contractile NM2 (nonmuscle myosin-2) motors, which help arrange actin stress fibers into thick ventral actin stress fibers. Using fluorescence anisotropy, we observed regional intracellular differences in myosin regulatory light chain phosphorylation in control LFs that were altered by dbn deletion. Using perturbations to induce and then release stalling of NM2 on actin in LFs from both genotypes, we made predictions explaining how DBN interacts with actin and NM2. These studies provide new insight for diseases such as emphysema and pulmonary fibrosis, in which fibroblasts inappropriately respond to mechanical cues in their environment.

Hyperoxia prevents the dynamic neonatal increases in lung mesenchymal cell diversity

Rapid expansion of the pulmonary microvasculature through angiogenesis drives alveolarization, the final stage of lung development that occurs postnatally and dramatically increases lung gas-exchange surface area. Disruption of pulmonary angiogenesis induces long-term structural and physiologic lung abnormalities, including bronchopulmonary dysplasia, a disease characterized by compromised alveolarization. Although endothelial cells are primary determinants of pulmonary angiogenesis, mesenchymal cells (MC) play a critical and dual role in angiogenesis and alveolarization. Therefore, we performed single cell transcriptomics and in-situ imaging of the developing lung to profile mesenchymal cells during alveolarization and in the context of lung injury. Specific mesenchymal cell subtypes were present at birth with increasing diversity during alveolarization even while expressing a distinct transcriptomic profile from more mature correlates. Hyperoxia arrested the transcriptomic progression of the MC, revealed differential cell subtype vulnerability with pericytes and myofibroblasts most affected, altered cell to cell communication, and led to the emergence of Acta1 expressing cells. These insights hold the promise of targeted treatment for neonatal lung disease, which remains a major cause of infant morbidity and mortality across the world.

The alveolus: Our current knowledge of how the gas exchange unit of the lung is constructed and repaired.

The mammalian lung completes its last step of development, alveologenesis, to generate sufficient surface area for gas exchange. In this process, multiple cell types that include alveolar epithelial cells, endothelial cells, and fibroblasts undergo coordinated cell proliferation, cell migration and/or contraction, cell shape changes, and cell-cell and cell-matrix interactions to produce the gas exchange unit: the alveolus. Full functioning of alveoli also involves immune cells and the lymphatic and autonomic nervous system. With the advent of lineage tracing, conditional gene inactivation, transcriptome analysis, live imaging, and lung organoids, our molecular understanding of alveologenesis has advanced significantly. In this review, we summarize the current knowledge of the constituents of the alveolus and the molecular pathways that control alveolar formation. We also discuss how insight into alveolar formation may inform us of alveolar repair/regeneration mechanisms following lung injury and the pathogenic processes that lead to loss of alveoli or tissue fibrosis.

Heterochrony and the evolution of the longjaw mudsucker (Gobiidae, Teleostei).

Although longjaw mudsucker (Gillichthys mirabilis, Gobiidae) has been studied extensively for its ability to occupy low-oxygen environments, few studies have addressed the evolution of its exceptionally elongated jaws that extend posteriorly beyond the gill opening in large adults. In this study, the ontogeny of the maxillae of G. mirabilis, Gillichthys seta, and the out-group species Eucyclogobius newberryi was studied within the heterochrony framework using digitized landmarks and caliper measurements. The results show that the maxilla of both species of Gillichthys evolved via acceleration (increased growth rate) and that of G. mirabilis via hypermorphosis (continued growth to a larger body size); two forms of peramorphosis. This is in contrast to earlier studies that concluded that G. seta is paedomorphic. We were unable to confirm an earlier hypothesis of sexual dimorphism in the jaw length of G. mirabilis. The evolution of the elongated jaws and associated large buccopharyngeal membrane in G. mirabilis is hypothesized to increase the surface area for gas exchange during aerial respiration and may also serve to amplify the aggressive gaping display as observed in other fishes.

The effect of heat on lung diffusing capacity for carbon monoxide (DLCO) during cycling exercise.

This study aimed to quantify the combined effects of heat exposure and exercise of increasing intensity on pulmonary blood flow using lung diffusing capacity for carbon monoxide (DLCO) as an indirect measure. We hypothesized that, during exercise in the heat, the well-documented increase in skin blood flow for thermoregulation would lead to alterations in pulmonary blood flow and a subsequent fall in DLCO versus a thermoneutral condition. Nine healthy subjects (4 F/5M, 20-45years, VO2max 46.7 ± 5.8mL/kg/min) completed three 15-min stages including rest and during cycling at 20 and 40% of maximum workload (Wmax) in either thermoneutral (TN; 22.2 ± 0.6°C) or heat (HT; 39.4 ± 0.4°C) conditions. DLCO, minute ventilation (VE), oxygen consumption ([Formula: see text]), heart rate (HR), and core (TC) and skin temperature (Tsk) were measured. DLCO showed a significant interaction between exercise intensity and heat (P = 0.019); post hoc testing revealed that DLCO was higher at 40% of Wmax in HT vs. TN (53.2 ± 10.6 vs 50.0 ± 10.3mL/min/mmHg, P = 0.003) only. VE and [Formula: see text] showed no difference in HT vs. TN. HR was higher in HT vs. TN (P < 0.001). TC and Tsk showed a significant interaction between temperature and intensity (P < 0.05). The unexpected increase in DLCO during exercise in HT vs. TN conditions suggests a larger lung surface area for gas exchange, perhaps due to increased pulmonary capillary recruitment and/or distension secondary to a higher cardiac output (Q) in the heat. This study furthers our understanding of how heat exposure might impact pulmonary blood flow, specifically as assessed via DLCO.

Asymmetric lung increases particle filtration by deposition

Human lung is known to be an asymmetric dichotomously branched network of bronchioles. Existing literature on the relation between anatomy and air-flow physics in the tracheobronchial trees has discussed the results of asymmetry. We discuss a secondary (but an important) lung function to seek asymmetry: to protect the acinus from a high pathogen load. We build morphometric parameter-based mathematical models of realistic bronchial trees to explore the structure-function relationship. We observe that maximum surface area for gas exchange, minimum resistance and minimum volume are obtained near the symmetry condition. In contrast, we show that deposition of inhaled foreign particles in the non-terminal airways is enhanced by asymmetry. We show from our model, that the optimal value of asymmetry for maximum particle filtration is within 10% of the experimentally measured value in human lungs. This structural trait of the lung aids in self-defence of the host against pathogen laden aerosols. We explain how natural asymmetric design of typical human lungs makes a sacrifice away from gas exchange optimality to gain this protection. In a typical human lung, when compared to most optimal condition (which is associated with symmetric branching), the fluidic resistance is 14% greater, the gas exchange surface area is about 11% lower, the lung volume is about 13% greater to gain an increase of 4.4% protection against foreign particles. This afforded protection is also robust to minor variations in branching ratio or variation in ventilation, which are both crucial to survival.

P44: Toward 3D-Printed Microfluidic Artificial Lungs with Complex, Biomimetic Geometries

Purpose: Current hollow-fiber oxygenators used in Extracorporeal Membrane Oxygenation have suboptimal gas exchange performance and hemocompatibility. Microfluidic artificial lungs (µALs) promise smaller feature sizes, improving gas exchange efficiency, and biomimetic flow paths, improving blood compatibility. For the first time, we leverage the geometric freedom of 3D-printing technology to demonstrate µALs with truly 3D, biomimetic branching capillaries and gas-side designs to increase gas exchange surface area. Methods: Two µAL designs were generated in Solidworks and printed on an Asiga MAX X27 UV printer. The first was a “simple branching” µAL (Fig 1A) with quadfurcating, biomimetic branching for blood distribution. The second was a “hatched” µAL (Fig 1B) employing a gas phase with strategically placed supports to maximize gas exchange surface area while maintaining structural stability. These changes are only to the gas, and so the blood-phase is unchanged. Dyed water was used to visualize internal structures (Fig 1D). Geometric analysis and proven models were used to calculate theoretical shear rate and rated flow. Computational Fluid Dynamics (CFD) was used to visualize flow (Fig 1E,F). Experimental pressure drop vs water flow rates were measured (Fig 2). Results: Through geometric analysis (Table 1), branching distribution technique provides more uniform shear stress (0.95-1.34 dyn/cm2) compared to a non-branching “open-plenum” control (0.09-1.27 dyn/cm2). The hatched µAL has a 3.18 mL/min rated flow, 1.7x greater than the 1.84 mL/min rated flow of the non-hatched branching µAL. As a result of the higher rated flow, it also has higher theoretical shear of 1.64-2.31 dyn/cm2 (at rated flow) despite identical blood-side geometry. The branching lung demonstrated a pressure drop up to 12 mmHg at flows up to 5 mL/min (Fig 2). These are significantly larger than analytically predicted values (extrapolated 7 mmHg vs 0.233 mmHg at 3.2 mL/min), prompting investigation into resolution of internal capillaries and effects of surface roughness on pressure drop. Conclusion: We have demonstrated the first 3D-printed branching biomimetic µAL formed from a gas permeable material, with gas-side geometries to improve gas exchange efficiency. These geometries are expected to translate to greater blood flows enabling creation of future animal and human-scale µALs. Table 1. Theoretical calculations. All uALs have 256 8.5mm long 216x324 µm blood and gas channels, with 54 µm-thick membranes. They differ in their effective gas exchange area, and blood distribution method, resulting in differing rated flows, shears, and pressure drops. The Branching uAL employs a branching distribution, resulting in a much tighter shear stress range. The GasHatch uAL employs both a branching distribution, and a blood-gas geometric-synchronization, with 100% rather than 58% of the gas-exchange capillary face being used for gas exchange. Values for a theoretical “control” lung (not fabricated; contains an open plenum/ non-branching distribution region) are displayed for comparison. Flow velocity and shear stresses are calculated separately for capillary and distribution areas.Figure 1. Blood flows are indicated using red arrows, and gas flows are indicated using blue arrows. (A) Design of simple (non-hatched) branched μAL. (B) Close-up of the hatched μAL capillaries and gas channels, showing how gas-side supports are only in regions not adjacent to capillaries and therefore not reducing gas exchange surface area. (C) Hatched μAL with the gas phase filled with blue dye for visualization. (D) Non-hatched branching μAL with blood phase filled with blue dye, and gas phase filled with red dye. (E) CFD visualization of flow velocities in the branching blood distribution network; (F) CFD analysis of pressure drop in the biomimetic blood distribution channels.Figure 2. Pressure drop of branching µALs (water; n=3)

BIO13: Effect of Artificial Lung Fiber Bundle Geometric Design on Micro- and Macro-scale Clot Formation

Purpose of Study: The hollow fiber membrane bundle is the functional component of artificial lungs, transferring oxygen and carbon dioxide to and from the blood. It is also the primary location of blood clot formation and propagation in these devices. The geometric design of fiber bundles is defined by a narrow range of parameters that determine gas exchange efficiency and blood flow resistance, such as fiber packing density, path length, and frontal area. However, these parameters also affect thrombosis. Methodology: This study investigated the effect of these parameters on clot formation using 3-D printed flow chambers that mimic the geometry and blood flow patterns of fiber bundles. Hollow fibers were represented by an array of vertical micro-rods (380 micron diameter) arranged with varying packing densities (40, 50, and 60%) and path lengths (2 and 4 cm), all of which are fiber bundle parameters in commercially available artificial lung devices. Blood was pumped through the device corresponding to three mean blood flow velocities (16, 20, and 25 cm/min), which were scaled from 4 L/min flowrates in an Adult Quadrox oxygenator. Results: Results showed that (1) clot formation decreases dramatically with decreasing packing density and increasing blood flow velocity, (2) clot formation at the outlet of fiber bundle enhances deposition upstream, and consequently (3) greater path length provides more clot-free fiber surface area for gas exchange than a shorter path length. Summary: These results can be used to create less thrombogenic, more efficient artificial lung designs.Figure 1. a) The experimental flow chamber was designed to be 2 or 4 cm and with a packing density of 60%, 50% or 40% rods to simulate the flow path of blood through an artificial lung hollow fiber bundle. (b) The flow circuit involved one-way flow from a syringe pump, connected to the flow chamber and then to a waste container. A manometer measured the pressure drop across each experimental device. (c) The averaged clot volumes from the microCT scans show more clot (red) at the outlets of all devices. There was also less clot in the chambers with a longer path length device, faster flow rate, and less density.

Discoidin domain receptor-2 enhances secondary alveolar septation in mice by activating integrins and modifying focal adhesions.

The extracellular matrix (ECM) of the pulmonary parenchyma must maintain the structural relationships among resident cells during the constant distortion imposed by respiration. This dictates that both the ECM and cells adapt to changes in shape, while retaining their attachment. Membrane-associated integrins and discoidin domain receptors (DDR) bind collagen and transmit signals to the cellular cytoskeleton. Although the contributions of DDR2 to collagen deposition and remodeling during osseous development are evident, it is unclear how DDR2 contributes to lung development. Using mice (smallie, Slie/Slie, DDR2Δ) bearing a spontaneous inactivating deletion within the DDR2 coding region, we observed a decrease in gas-exchange surface area and enlargement of alveolar ducts. Compared with fibroblasts isolated from littermate controls, DDR2Δ fibroblasts, spread more slowly, developed fewer lamellipodia, and were less responsive to the rigidity of neighboring collagen fibers. Activated β1-integrin (CD29) was reduced in focal adhesions (FA) of DDR2Δ fibroblasts, less phospho-zyxin localized to and fewer FA developed over ventral actin stress fibers, and the adhesions had a lower aspect ratio compared with controls. However, DDR2 deletion did not reduce cellular displacement of the ECM. Our findings indicate that DDR2, in concert with collagen-binding β1-integrins, regulates the timing and location of focal adhesion formation and how lung fibroblasts respond to ECM rigidity. Reduced rigidity sensing and mechano-responsiveness may contribute to the distortion of alveolar ducts, where the fiber cable-network is enriched and tensile forces are concentrated. Strategies targeting DDR2 could help guide fibroblasts to locations where tensile forces organize parenchymal repair.

The Relationship Between the Weight of the Placenta and Birth Weight of the Neonate

INTRODUCTION Weight of placenta has long been associated with fetal growth and perinatal well-being. Low placental weight (standardized for sex and gestational age) is more common among stillbirths whereas high placental weight is associated with neonatal death, serious neonatal morbidity, low 5-min Apgar score, and respiratory morbidity. Low placental weight is believed to reflect an inadequate placental surface area for nutrient and gas exchange, leading to fetal compromise. However, it has also been proposed that the increased risk associated with high placental weight is the result of villous oedema, which may compress blood vessels and reduce transfer of gas and nutrients to the fetus. MATERIAL AND METHODS A descriptive cross-sectional study of the total, 798 subjects met the inclusive criteria. After consent, age, sex, prepregnancy weight, weight of mother at the time of delivery, neonatal weight and trimmed placental weight were recorded. RESULTS The mean birth weight was 2818±447.19 gm while the placental weight was 489.29±59 gm with mean placental-fetal weight ratio of 17.63. The mean birthweight of male was 2823.97±451.12 gm whereas female was 2811.69±442.93 gm. Similarly, mean placental weight of male was 490.37±57.7 gm and female was 487.98±60.58 gm. As gestational age increases placental and birth weight also increases whereas placental-birth weight ratio decreases. CONCLUSION There is a positive relation between placental weight and weight of the neonate at the time of birth. However, the ratio of the placental and neonatal birth weights at term decreases with advancing gestational age. Thus, prolongation of pregnancy at term may adversely affect the fetus.

A hybrid discrete-continuum model of immune responses to SARS-CoV-2 infection in the lung alveolar region, with a focus on interferon induced innate response

We develop a lattice-based, hybrid discrete-continuum modeling framework for SARS-CoV-2 exposure and infection in the human lung alveolar region, or parenchyma, the massive surface area for gas exchange. COVID-19 pneumonia is alveolar infection by the SARS-CoV-2 virus significant enough to compromise gas exchange. The modeling framework orchestrates the onset and progression of alveolar infection, spatially and temporally, beginning with a pre-immunity baseline, upon which we superimpose multiple mechanisms of immune protection conveyed by interferons and antibodies. The modeling framework is tunable to individual profiles, focusing here on degrees of innate immunity, and to the evolving infection–replication properties of SARS-CoV-2 variant strains. The model employs partial differential equations for virion, interferon, and antibody concentrations governed by diffusion in the thin fluid coating of alveolar cells, species and lattice interactions corresponding to sources and sinks for each species, and multiple immune protections signaled by interferons. The spatial domain is a two-dimensional, rectangular lattice of alveolar type I (non-infectable) and type II (infectable) cells with a stochastic, species-concentration-governed, switching dynamics of type II lattice sites from healthy to infected. Once infected, type II cells evolve through three phases: an eclipse phase during which RNA copies (virions) are assembled; a shedding phase during which virions and interferons are released; and then cell death. Model simulations yield the dynamic spread of, and immune protection against, alveolar infection and viral load from initial sites of exposure. We focus in this paper on model illustrations of the diversity of outcomes possible from alveolar infection, first absent of immune protection, and then with varying degrees of four known mechanisms of interferon-induced innate immune protection. We defer model illustrations of antibody protection to future studies. Results presented reinforce previous recognition that interferons produced solely by infected cells are insufficient to maintain a high efficacy level of immune protection, compelling additional mechanisms to clear alveolar infection, such as interferon production by immune cells and adaptive immunity (e.g., T cells). This manuscript was submitted as part of a theme issue on “Modelling COVID-19 and Preparedness for Future Pandemics”.

Impact of fetal treatments for congenital diaphragmatic hernia on lung development.

The extent of lung hypoplasia impacts the survival and severity of morbidities associated with congenital diaphragmatic hernia (CDH). The alveoli of CDH infants and in experimental models of CDH have thickened septa with fewer type II pneumocytes and capillaries. Fetal treatments of CDH-risk preterm birth. Therefore, treatments must aim to balance the need for increased gas exchange surface area with the restoration of pulmonary epithelial type II cells and the long-term respiratory and neurodevelopmental consequences of prematurity. Achievement of sufficient lung development in utero for successful postnatal transition requires adequate intra-thoracic space for lung growth, maintenance of sufficient volume and appropriate composition of fetal lung fluid, regular fetal breathing movements, appropriate gas exchange area, and ample surfactant production. The review aims to examine the rationale for current and future therapeutic strategies to improve postnatal outcomes of infants with CDH.

A functional circuit formed by the autonomic nerves and myofibroblasts controls mammalian alveolar formation for gas exchange.

Alveolar formation increases the surface area for gas exchange. A molecular understanding of alveologenesis remains incomplete. Here, we show that the autonomic nerve and alveolar myofibroblast form a functional unit in mice. Myofibroblasts secrete neurotrophins to promote neurite extension/survival, whereas neurotransmitters released from autonomic terminals are necessary for myofibroblast proliferation and migration, a key step in alveologenesis. This establishes a functional link between autonomic innervation and alveolar formation. We also discover that planar cell polarity (PCP) signaling employs a Wnt-Fz/Ror-Vangl cascade to regulate the cytoskeleton and neurotransmitter trafficking/release from the terminals of autonomic nerves. This represents a new aspect of PCP signaling in conferring cellular properties. Together, these studies offer molecular insight into how autonomic activity controls alveolar formation. Our work also illustrates the fundamental principle of how two tissues (e.g., nerves and lungs) interact to build alveoli at the organismal level.

Interaction of epithelium and myofibroblasts through ERBB3‐PDGF growth pathways crosstalk drive the maturation of distal lung epithelium in late stage lung development

RationaleThe last stage of lung development is realized by coordinated growth of septae from the alveolar walls to subdivide the distal saccules into alveoli, resulting in the expansion of surface area for gas exchange. The process is accompanied by microvascular maturation and underlying matrix formation of the interstitial scaffold. Secondary septation is driven by the orchestration of the distal epithelium, endothelium, and fibroblasts through the spatial and temporal distribution of various growth pathways and their crosstalk.This study investigated the role of the receptor tyrosine kinases 3 (ERBB3) pathway in the fibroblast‐epithelial crosstalk during septation using both in vitro co‐cultures and ex vivo organoid model as well as transgenic mice.MethodsPrimary myofibroblasts were extracted from neonatal C57BL/6wildtype (WT) mice at postnatal day 7 (PND7) and co‐cultured with a murine epithelial cell line (MLE12) in transwell inserts for 24h before sample collection. Gene expression was analyzed by quantitative PCR. Neonatal lung organoids were generated from primary lung epithelial cells (CD45‐/CD31‐) from neonatal C57BL/6 WT mice (PND7) and cultured in matrigel together with primary myofibroblasts from neonatal WT mice (PND7) as well as age matched mice heterozygot for the Platelet‐derived growth factor receptors (PDGF‐Rα+/‐). After organoid formation, cultures were treated with Neuregulin (NRG), an activating ligand of the ERBB3 receptor. We measured organoid number and size and Erbb3 signaling was assessed by immunofluorescence and quantitative gene expression analyses.ResultsProximal culture with myofibroblasts upregulated the expression level of ERBB3 receptor in epithelial cells, as well as homeodomain‐only protein homeobox (HOPX), GATA‐binding factor 6 (GATA6), both of which are essential in the differentiation of distal epithelial cells and surfactant production during late stage lung development. In neonatal lung organoids, myofibroblasts with reduced PDGFRα expression led to reduction in organoid size and number. Treatment with the Erbb3 ligand NRG significantly increased both the expression and phosphorylation of Erbb3 in epithelial cells together with an upregulation of surfactant C (SPC) expression, and an increase in myofibroblasts PDGF‐R α expression.ConclusionActivated Erbb3 signaling results from the interaction with fibroblasts involving PDGF‐Rα signaling and drives distal epithelium maturation.

Morphophysiological aspects of ornamental sunflowers cultivated in different growing seasons under semi-arid conditions

ABSTRACT Knowledge of how climatic conditions affect plant morphophysiology is essential for understanding how to manage the growth cycles of different crops. The aim of this study was to evaluate the effects of the growing seasons in a semi-arid area on the morphophysiological variables of ornamental sunflower plants. The experiment was carried out in a randomized block design in a split-plot arrangement with four replicates. Six cultivars (‘Bonito de Outono Sortido’, ‘Sol Noturno’, ‘Sol Vermelho’, ‘Jardim Amarelo Alto’, ‘Girassol F1 Sunbright Supreme’ and ‘Girassol F1 Vincents Choice’) were evaluated in the main plots and two different growing seasons (GS) in the subplots (GS1 - warm climate and GS2 - mild climate). Evaluations of gas exchange, chlorophyll indices, and leaf surface area were carried out at the reproductive stage (R5.5). The cultivation of ornamental sunflowers in semi-arid regions was significantly affected by the growing season. Changes in gas exchange variables and the morphophysiology of ornamental sunflower plants in the two growing seasons reflected the high phenotypic plasticity characteristic of this species. The cultivation of ornamental sunflowers under semi-arid conditions in the growing season, when air temperature and solar radiation are high, could be limited due to elevated transpiration rates. Therefore, it is recommended that they are grown mainly during the moderate climatic season in semi-arid regions.

Microstructured Hollow Fiber Membranes: Potential Fiber Shapes for Extracorporeal Membrane Oxygenators.

Extracorporeal membrane oxygenators are essential medical devices for the treatment of patients with respiratory failure. A promising approach to improve oxygenator performance is the use of microstructured hollow fiber membranes that increase the available gas exchange surface area. However, by altering the traditional circular fiber shape, the risk of low flow, stagnating zones that obstruct mass transfer and encourage thrombus formation, may increase. Finding an optimal fiber shape is therefore a significant task. In this study, experimentally validated computational fluid dynamics simulations were used to investigate transverse flow within fiber packings of circular and microstructured fiber geometries. A numerical model was applied to calculate the local Sherwood number on the membrane surface, allowing for qualitative comparison of gas exchange capacities in low-velocity areas caused by the microstructured geometries. These adverse flow structures lead to a tradeoff between increased surface area and mass transfer. Based on our simulations, we suggest an optimal fiber shape for further investigations that increases potential mass transfer by up to 48% in comparison to the traditional, circular hollow fiber shape.

Age-dependent alveolar epithelial plasticity orchestrates lung homeostasis and regeneration.

Regeneration of the architecturally complex alveolar niche of the lung requires precise temporal and spatial control of epithelial cell behavior. Injury can lead to a permanent reduction in gas exchange surface area and respiratory function. Using mouse models, we show that alveolar type 1 (AT1) cell plasticity is a major and unappreciated mechanism that drives regeneration, beginning in the early postnatal period during alveolar maturation. Upon acute neonatal lung injury, AT1 cells reprogram into alveolar type 2 (AT2) cells, promoting alveolar regeneration. In contrast, the ability of AT2 cells to regenerate AT1 cells is restricted to the mature lung. Unbiased genomic assessment reveals that this previously unappreciated level of plasticity is governed by the preferential activity of Hippo signaling in the AT1 cell lineage. Thus, cellular plasticity is a temporally acquired trait of the alveolar epithelium and presents an alternative mode of tissue regeneration in the postnatal lung.