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Abstract The lymphatic system consists of a vessel network lined by specialized lymphatic endothelial cells (LECs) that are responsible for tissue fluid homeostasis and immune cell trafficking. The mechanisms for organ-specific LEC responses to environmental cues are not well understood. We found robust lymphangiogenesis during influenza A virus infection in the adult mouse lung. We show that the number of LECs increases twofold at 7 days post-influenza infection (dpi) and threefold at 21 dpi, and that lymphangiogenesis is preceded by lymphatic dilation. We also show that the expanded lymphatic network enhances fluid drainage to mediastinal lymph nodes. Using EdU labeling, we found that a significantly higher number of pulmonary LECs are proliferating at 7 dpi compared to LECs in homeostatic conditions. Lineage tracing during influenza indicates that new pulmonary LECs are derived from preexisting LECs rather than non-LEC progenitors. Lastly, using a conditional LEC-specific YAP/TAZ knockout model, we established that lymphangiogenesis, fluid transport and the immune response to influenza are independent of YAP/TAZ activity in LECs. These findings were unexpected, as they indicate that YAP/TAZ signaling is not crucial for these processes.

Summary: Macrophages are critical for maintenance and repair of mucosal tissues. While functionally distinct subtypes of macrophage are known to have important roles in injury response and repair in the lungs, little is known about macrophages in the proximal conducting airways. Single-cell RNA sequencing and flow cytometry demonstrated murine tracheal macrophages are largely monocyte-derived and are phenotypically distinct from lung macrophages at homeostasis. Following sterile airway injury, monocyte-derived macrophages are recruited to the trachea and activate a pro-regenerative phenotype associated with wound healing. Animals lacking the chemokine receptor CCR2 have reduced numbers of circulating monocytes and tracheal macrophages, deficient pro-regenerative macrophage activation and defective epithelial repair. Together, these studies indicate that recruitment and activation of monocyte-derived tracheal macrophages is CCR2-dependent and is required for normal airway epithelial regeneration.

The respiratory system, which includes the trachea, airways, and distal alveoli, is a complex multi-cellular organ that intimately links with the cardiovascular system to accomplish gas exchange. In this review and as members of the NIH/NHLBI-supported Progenitor Cell Translational Consortium, we discuss key aspects of lung repair and regeneration. We focus on the cellular compositions within functional niches, cell-cell signaling in homeostatic health, the responses to injury, and new methods to study lung repair and regeneration. We also provide future directions for an improved understanding of the cell biology of the respiratory system, as well as new therapeutic avenues.

The lymphatic system consists of a vessel network lined by specialized lymphatic endothelial cells (LECs) that are responsible for tissue fluid homeostasis and immune cell trafficking. The mechanisms for organ-specific LEC responses to environmental cues are not well understood. We found robust lymphangiogenesis during influenza A virus infection in the adult mouse lung. We show that the number of LECs increases twofold at 7 days post-influenza infection (dpi) and threefold at 21 dpi, and that lymphangiogenesis is preceded by lymphatic dilation. We also show that the expanded lymphatic network enhances fluid drainage to mediastinal lymph nodes. Using EdU labeling, we found that a significantly higher number of pulmonary LECs are proliferating at 7 dpi compared to LECs in homeostatic conditions. Lineage tracing during influenza indicates that new pulmonary LECs are derived from preexisting LECs rather than non-LEC progenitors. Lastly, using a conditional LEC-specific YAP/TAZ knockout model, we established that lymphangiogenesis, fluid transport and the immune response to influenza are independent of YAP/TAZ activity in LECs. These findings were unexpected, as they indicate that YAP/TAZ signaling is not crucial for these processes. [ABSTRACT FROM AUTHOR]

The presence of green roofs in urban areas improves the energy efficiency of buildings; contributes to the capture of CO2, decreasing pollution; and improves the appearance of cities, increasing their sustainability. Additionally, green roofs must include plant species with low requirements and maintenance, and thus, succulent species could be useful in Mediterranean semi-arid regions. In this work, the thermal inertia and the effect of different succulent species as thermal barriers on mitigating the increase in temperature inside experimental compartments were studied in comparison to conventional covers used in the Spanish Mediterranean for a whole year. In general, green covers were more efficient than conventional ones for controlling temperature. Thus, temperatures under green covers were up to 8 °C lower than conventional covers and 3–5 °C lower than the ambient temperature at noon on summer days. Furthermore, significant differences were found between green covers. Thus, despite having high thermal inertia, Aptenia cordifolia showed the worst temperature records, while Aeonium arboreum was the most efficient at mitigating temperature changes both on cold winter nights and hot summer days—even better than Sedum spp., a usual succulent used commercially. Our results demonstrate that succulent species are efficient materials to use as green covers to improve thermal conditions in buildings in Mediterranean cities. This also suggests that the mixture of succulent species (i.e., not only made of Sedum spp.) with different colors and textures could beautify green roofs without compromising their energy efficiency.

Lung regeneration occurs in a variety of adult mammals after surgical removal of one lung (pneumonectomy). Previous studies of murine post-pneumonectomy lung growth have identified regenerative “hotspots” in subpleural alveolar ducts; however, the cell-types participating in this process remain unclear. To identify the single cells participating in post-pneumonectomy lung growth, we used laser microdissection, enzymatic digestion and microfluidic isolation. Single-cell transcriptional analysis of the murine alveolar duct cells was performed using the C1 integrated fluidic circuit (Fluidigm) and a custom PCR panel designed for lung growth and repair genes. The multi-dimensional data set was analyzed using visualization software based on the tSNE algorithm. The analysis identified 6 cell clusters; 1 cell cluster was present only after pneumonectomy. This post-pneumonectomy cluster was significantly less transcriptionally active than 3 other clusters and may represent a transitional cell population. A provisional cluster identity for 4 of the 6 cell clusters was obtained by embedding bulk transcriptional data into the tSNE analysis. The transcriptional pattern of the 6 clusters was further analyzed for genes associated with lung repair, matrix production, and angiogenesis. The data demonstrated that multiple cell-types (clusters) transcribed genes linked to these basic functions. We conclude that the coordinated gene expression across multiple cell clusters is likely a response to a shared regenerative microenvironment within the subpleural alveolar ducts.

La dextrocardia es una rara anomalía cardiaca congénita. Aunque la incidencia de enfermedad coronaria es similar a la población en general, son pocos los casos documentados en la literatura. La disposición anatómica en esta anomalía constituye un desafío a la hora de planificar la técnica quirúrgica y los injertos a utilizar, como lo describen los casos presentados. Presentamos el caso de un paciente varón de 73 años de edad, con enfermedad coronaria de 2 vasos que fue sometido a una cirugía de revascularización arterial completa, sin circulación extracorpórea y sin modificaciones significativas de la técnica convencional.

ObjectivesThe mesothelium, the surface layer of the heart, lung, bowel, liver, and tunica vaginalis, is a complex tissue implicated in organ-specific diseases and regenerative biology; however, the mechanism of mesothelial repair after surgical injury is unknown. Previous observations indicated seeding of denuded mesothelium by free-floating mesothelial cells may contribute to mesothelial healing. In this study, we investigated the prevalence of mesothelial cells in pleural fluid during the 7 days following pulmonary surgery.Study designFlow cytometry was employed to study pleural fluid of 45 patients after lung resection or transplantation. We used histologically validated mesothelial markers (CD71 and WT1) to estimate the prevalence of mesothelial cells.ResultsThe viability of pleural fluid cells approached 100%. Leukocytes and mesothelial cells were identified in the pleural fluid within the first week after surgery. The leukocyte concentration was relatively stable at all time points. In contrast, mesothelial cells, identified by CD71 and WT1 peaked on POD3. The broad expression of CD71 molecule in postoperative pleural fluid suggests that many of the free-floating non-leukocyte cells were activated or proliferative mesothelial cells.ConclusionWe demonstrated that pleural fluid post lung surgery is a source of mesothelial cells; most of these cells appear to be viable and, as shown by CD71 staining, activated mesothelial cells. The observed peak of mesothelial cells on POD3 is consistent with a potential reparative role of free-floating mesothelial cells after pulmonary surgery.

In many mammals, including rodents and humans, removal of one lung results in the compensatory growth of the remaining lung; however, the mechanism of compensatory lung growth is unknown. Here, we investigated the changes in morphology and phenotype of pleural cells after pneumonectomy. Between days 1 and 3 after pneumonectomy, cells expressing α-smooth muscle actin (SMA), a cytoplasmic marker of myofibroblasts, were significantly increased in the pleura compared to surgical controls (p < .01). Scanning electron microscopy of the pleural surface 3 days post-pneumonectomy demonstrated regions of the pleura with morphologic features consistent with epithelial-mesenchymal transition (EMT); namely, cells with disrupted intercellular junctions and an acquired mesenchymal (rounded and fusiform) morphotype. To detect the migration of the transitional pleural cells into the lung, a biotin tracer was used to label the pleural mesothelial cells at the time of surgery. By post-operative day 3, image cytometry of post-pneumonectomy subpleural alveoli demonstrated a 40-fold increase in biotin+ cells relative to pneumonectomy-plus-plombage controls (p < .01). Suggesting a similar origin in space and time, the distribution of cells expressing biotin, SMA, or vimentin demonstrated a strong spatial autocorrelation in the subpleural lung (p < .001). We conclude that post-pneumonectomy compensatory lung growth involves EMT with the migration of transitional mesothelial cells into subpleural alveoli.

Glasshouses in Mediterranean areas are mainly used for experimental or nursery purposes and the production of high value ornamental species. In addition, glasshouses may be covered by dust and other particles in these areas as rainfalls are relatively scarce. Consequently, the quality of photosynthetically active radiation (PAR) may be affected. In the present experiment we evaluated the effect of dirt on the PAR spectrum in glasshouses in Valencia (Spain), compared to glasshouses submitted to suitable maintenance and washing. Photosynthetic Photon Flux Density (PPFD) measurements were done in March 2017 at 13h, and visible, UV-C and near infrared regions were recorded with a portable spectrometer. PPFD maximum values differed depending on the spectral region and glass conditions, with higher values corresponding to the visible region. Thus, clean glass showed values of 609, 588 and 423 μmol/m2s for PPFDR, PPFDG and PPFDB, respectively and 436 and 34 μmol/m2s for PPFDIR and PPFDUV. In comparison to clean glass, dirty glass decreased PPFD in all regions, particularly UV and blue region.

Tipa (Tipuana tipu) is a common tree in gardens and carparks, although the shading effect of its canopy must be still characterised to assess the decrease of temperature and quality of irradiance. This work is a preliminary study aimed to assess the effect of shading of tree canopies on the diffuse irradiance pattern received at the soil level in comparison to other conditions. The shade provided by a group of Tipa trees, located at the Universitat Politècnica de València (Valencia, Spain), was evaluated in this experiment and compared to cloudy days and direct sun. Photosynthetic Photon Flux Density (PPFD) and red/near infrared ratios were recorded with a portable spectrometer. Measurements were recorded in January and February 2017, at 10h, 13h and 16h. Depending on the region of the spectrum and time of the day, PPFD values ranged from 0.05 to 0.42, 0.40 to 1.14, and 0.94 to 3.90 μmol·m-2·s-1, for Tipa shade, cloudy days and direct sun, respectively. The spectral analysis of PPFD in cloudy days revealed maximum values in the green region and minimum at near infrared region, while maximum PPFD for tipa shade was mostly found at near infrared, revealing higher importance of this spectral region compared to cloudy days.