The role of vasculature and angiogenesis in respiratory diseases.

In European countries, nearly 10% of all hospital admissions are related to respiratory diseases, mainly chronic life-threatening diseases such as COPD, pulmonary hypertension, IPF or lung cancer. The contribution of blood vessels and angiogenesis to lung regeneration, remodeling and disease progression has been increasingly appreciated. The vascular supply of the lung shows the peculiarity of dual perfusion of the pulmonary circulation (vasa publica), which maintains a functional blood-gas barrier, and the bronchial circulation (vasa privata), which reveals a profiled capacity for angiogenesis (namely intussusceptive and sprouting angiogenesis) and alveolar-vascular remodeling by the recruitment of endothelial precursor cells. The aim of this review is to outline the importance of vascular remodeling and angiogenesis in a variety of non-neoplastic and neoplastic acute and chronic respiratory diseases such as lung infection, COPD, lung fibrosis, pulmonary hypertension and lung cancer.

Age-related changes in testicular histological structure, testosterone level, libido and semen traits in male Coturnix quail.

To investigate the effect of age in male quail on testicular weight and histology, sexual libido and semen characteristics, a study was performed on 100 quails at 10, 16, 22, 28 and 34 weeks of age. The body and testicular weights were significantly (p < .05) higher at 16 and 22 than at 28 weeks of age. The circumference and diameter of the seminiferous tubules were significantly (p < .05) higher at 28 and 34 than at 10 and 16 weeks of age. Histological evaluation of testicular slices revealed advanced and effective seminiferous tubes as early as 10 weeks, while spermatogenic activity peaked at 16 weeks of age. The highest semen volume, sperm motility and sperm concentration were observed at weeks of age and then decreased gradually with age. At the same time, the testosterone level and libido were significantly (p < .05) higher at 22 than at 10 weeks of age. Furthermore, the age was positive (p < .05), correlated to seminiferous tubule circumference and diameter, and negative (p < .05), correlated to sperm concentration. In conclusion, the age of quail markedly affected the testicular histological structure, libido, testosterone level and semen characteristics.

Evolving topological order in the postnatal visceral pleura.

BACKGROUND: Changes in epithelial cell shape reflects optimal cell packing and the minimization of surface free energy, but also cell-cell interactions, cell proliferation, and cytoskeletal rearrangements. RESULTS: Here, we studied the structure of the rat pleura in the first 15 days after birth. After pleural isolation and image segmentation, the analysis demonstrated a progression of epithelial order from postnatal day 1 (P1) to P15. The cells with the largest surface area and greatest shape variability were observed at P1. In contrast, the cells with the smallest surface area and most shape consistency were observed at P15. A comparison of polygonal cell geometries demonstrated progressive optimization with an increase in the number of hexagons (six-sided) as well as five-sided and seven-sided polygons. Analysis of the epithelial organization with Voronoi tessellations and graphlet motif frequencies demonstrated a developmental path strikingly distinct from mathematical and natural reference paths. Graph Theory analysis of cell connectivity demonstrated a progressive decrease in network heterogeneity and clustering coefficient from P1 to P15. CONCLUSIONS: We conclude that the rat pleura undergoes a striking change in pleural structure from P1 to P15. Further, a geometric and network-based approach can provide a quantitative characterization of these developmental changes.

Vascularization of the adult mouse lung grafted onto the chick chorioallantoic membrane.

In the later stages of angiogenesis, the vascular sprout transitions into a functional vessel by fusing with a target vessel. Although this process appears to routinely occur in embryonic tissue, the biologic rules for sprout fusion and lumenization in adult regenerating tissue are unknown. To investigate this process, we grafted portions of the regenerating post-pneumonectomy lung onto the chick chorioallantoic membrane (CAM). Grafts from all 4 lobes of the post-pneumonectomy right lung demonstrated peri-graft angiogenesis as reflected by fluorescent plasma markers; however, fluorescent microsphere perfusion primarily occurred in the lobe of the lung that is the dominant site of post-pneumonectomy angiogenesis-namely, the cardiac lobe. Vascularization of the cardiac lobe grafts was confirmed by active tissue growth (p < .05). Functional vascular connections between the cardiac lobe and the CAM vascular network were demonstrated by confocal fluorescence microscopy as well as corrosion casting and scanning electron microscopy (SEM). Bulk transcriptional profiling of the cardiac lobe demonstrated the enhanced expression of many genes relative to alveolar epithelial cell (CD11b-/CD31-) control cells, but only the upregulation of Ereg and Fgf6 compared to the less well-vascularized right upper lobe. The growth of actively regenerating non-neoplastic adult tissue on the CAM demonstrates that functional lumenization can occur between species (mouse and chick) and across the developmental spectrum (adult and embryo).

Influence of dietary mannan-oligosaccharides supplementation on hematological characteristics, blood biochemical parameters, immune response and histological state of laying hens.

This study aimed to determine the influence of dietary mannan-oligosaccharides (MOS) on the immune system, hematological traits, blood biochemical parameters, and histological state of laying hens. At 34 wk of age, The Mandarah chicken strain's 120 laying hens and 12 cocks were divided into 4 groups, each with 30 hens and 3 cocks. The first group performed as a control group, which nourished on a basal diet. The second, third, and fourth experimental groups received 0.1, 0.2, and 0.5 g/kg of MOS and a base diet, respectively. Birds obtained MOS at numerous doses significantly (P ˂ 0.05) raised serum levels of immunoglobulin Y (IgY), immunoglobulin M (IgM), and avian influenza (AI) antibodies compared to control birds. Furthermore, adding MOS at a level of 0.1 g/kg diet significantly improved the immune response of the control group. Additionally, compared to the control group, treated birds with MOS at various dosages did not significantly enhance hematological parameters such as red blood cells (RBCs), white blood cells (WBCs), hemoglobin, and hematocrit. Compared to control birds, birds fed MOS at all levels exhibited considerably lower serum cholesterol, alanine aminotransferase (ALT), and aspartate aminotransferase (AST) values. Also, compared to other treated birds, MOS-treated birds displayed improved histological examination of the small intestine, isthmus, and testis compared to the control group, particularly in birds fed MOS at 0.1 and 0.2 g/kg diet. It could be concluded that using MOS at 0.1 or 2 g/kg diet can successfully improve the physiological performance and overall health of laying hens.

Immune response, hematological traits, biochemical blood parameters, and histological status of laying hens influenced by dietary chitosan-oligosaccharides.

This experiment aimed to examine the effect of chitosan-oligosaccharides (COS) supplementation in laying hens' diets affected their immune response, hematological characteristics, blood biochemical parameters, and histological status. At the age of 34 wk, 200 laying hens and 20 cocks of the Mandarah chicken strain were allotted into four groups, each consisting of 50 hens and five cocks. The first group acted as a control group, fed on a basal diet. The second, third, and fourth experimental groups each received 0.1, 0.2, and 0.5 g/kg of COS in addition to a base diet. Birds received COS at various dosages had significantly (P ˂ 0.05) increased serum concentration of immunoglobulins, avian influenza, and Newcastle disease antibodies compared with the control birds. Moreover, adding COS at level 0.2 g/kg diet insignificantly enhanced immune response than the rest of the treatment groups. Also, treated birds with COS at different levels had insignificantly improved hematological parameters such as red blood cells, white blood cells, hemoglobin and hematocrit compared to the control group. Birds fed COS at all levels had significantly decreased serum cholesterol, triglycerides, Ca++ and alanine aminotransferase concentrations compared with control birds. In addition, compared to the control group, chitosan-treated birds showed enhanced histological examination of the small intestine, isthmus, and testis, notably in birds given COS at 0.1 g/kg diet compared to other treated birds. Cocks fed COS at all levels improved testicular tissues and increased the number and diameter of seminiferous tubules compared with control birds Morphological examination of the ileum showed increased villi number, height, and crypt depth. It is possible to conclude that laying hens' physiological performance and general health can be effectively improved by using chitosan at 0.1 or 2 g/kg diet levels enhanced immune response.

Biomechanics of a Plant-Derived Sealant for Corneal Injuries.

Purpose: The corneal epithelium has a glycocalyx composed of membrane-associated glycoproteins, mucins, and galactin-3. Similar to the glycocalyx in visceral tissues, the corneal glycocalyx functions to limit fluid loss and minimize frictional forces. Recently, the plant-derived heteropolysaccharide pectin has been shown to physically entangle with the visceral organ glycocalyx. The ability of pectin to entangle with the corneal epithelium is unknown. Methods: To explore the potential role of pectin as a corneal bioadhesive, we assessed the adhesive characteristics of pectin films in a bovine globe model. Results: Pectin film was flexible, translucent, and low profile (80 µm thick). Molded in tape form, pectin films were significantly more adherent to the bovine cornea than control biopolymers of nanocellulose fibers, sodium hyaluronate, and carboxymethyl cellulose (P < 0.05). Adhesion strength was near maximal within seconds of contact. Compatible with wound closure under tension, the relative adhesion strength was greatest at a peel angle less than 45 degrees. The corneal incisions sealed with pectin film were resistant to anterior chamber pressure fluctuations ranging from negative 51.3 ± 8.9 mm Hg to positive 214 ± 68.6 mm Hg. Consistent with these findings, scanning electron microscopy demonstrated a low-profile film densely adherent to the bovine cornea. Finally, the adhesion of the pectin films facilitated the en face harvest of the corneal epithelium without physical dissection or enzymatic digestion. Conclusions: We conclude that pectin films strongly adhere to the corneal glycocalyx. Translational Relevance: The plant-derived pectin biopolymer provides potential utility for corneal wound healing as well as targeted drug delivery.

Geometric and network organization of visceral organ epithelium.

Mammalian epithelia form a continuous sheet of cells that line the surface of visceral organs. To analyze the epithelial organization of the heart, lung, liver and bowel, epithelial cells were labeled in situ, isolated as a single layer and imaged as large epithelial digitally combine montages. The stitched epithelial images were analyzed for geometric and network organization. Geometric analysis demonstrated a similar polygon distribution in all organs with the greatest variability in the heart epithelia. Notably, the normal liver and inflated lung demonstrated the largest average cell surface area (p < 0.01). In lung epithelia, characteristic wavy or interdigitated cell boundaries were observed. The prevalence of interdigitations increased with lung inflation. To complement the geometric analyses, the epithelia were converted into a network of cell-to-cell contacts. Using the open-source software EpiGraph, subgraph (graphlet) frequencies were used to characterize epithelial organization and compare to mathematical (Epi-Hexagon), random (Epi-Random) and natural (Epi-Voronoi5) patterns. As expected, the patterns of the lung epithelia were independent of lung volume. In contrast, liver epithelia demonstrated a pattern distinct from lung, heart and bowel epithelia (p < 0.05). We conclude that geometric and network analyses can be useful tools in characterizing fundamental differences in mammalian tissue topology and epithelial organization.

Analysis of recurrence in lung adenocarcinoma with spread through air spaces.

OBJECTIVES: Spread through air spaces is defined as tumor cells in air spaces away from the edge of tumor in lung carcinoma. It is associated with higher locoregional recurrence and lower survival in lung adenocarcinoma. The features of spread through air spaces portending worse outcomes are still under investigation. We reviewed our lung cancer experience to define potential factors related to spread through air spaces that influence recurrence and survival. METHODS: Between January 2010 and December 2017, we identified 968 patients who underwent resection for T1-3N0M0 lung adenocarcinoma. Of these, histologic examination was possible in 787 patients. We examined the presence of spread through air spaces, spread through air spaces characteristics (micropapillary, solid nest, or single cell), average density (number per slide), and farthest distance from tumor at which spread through air spaces was detected, or maximal spread distance. Overall survival and recurrence-free survival were estimated using Kaplan-Meier curves, and differences between spread through air spaces positive versus spread through air spaces negative groups were assessed using the log-rank test. RESULTS: Spread through air spaces was present in 389 of 787 of the reviewed cases (49.4%). Overall survival and recurrence-free survival were significantly lower in the spread through air spaces positive group over 10 years (P < .0001). The incidences of locoregional and distant recurrence were nearly doubled over 10 years in the spread through air spaces positive group compared with the spread through air spaces negative group (P = .002 and <.0001, respectively). In a multivariable Cox regression model adjusted for spread through air spaces characteristics, distance, and tumor size, lobar resection did not confer survival advantage in patients with spread through air spaces (hazard ratio of sublobar resection with respect to lobar resection, 1.44; 95% confidence interval, 0.98-2.11; P = .067). In the spread through air spaces positive group, spread through air spaces density was 2.7 ± 1.4 clusters per slide and the maximal spread distance was 2.2 ± 1.7 mm from the tumor edge. There was no observed correlation between spread through air spaces density or maximal spread distance and overall survival or recurrence. CONCLUSIONS: We show increased distant recurrence in spread through air spaces positive lung adenocarcinoma. Quantifiable measures of spread through air spaces do not appear to correlate with recurrence or survival metrics.

Effect of dietary chitosan supplementation on productive and physiological performance parameters of growing New Zealand white rabbits.

This study aimed to investigate the effects of dietary chitosan supplementation on the growth performance, carcass traits, and some physiological parameters of weaned New Zealand White rabbits (NZW). Sixty-four NZWs were randomly distributed into four dietary experimental groups (n = 16) and fed a basal diet ad libitum for eight weeks: under Egyptian conditions. The basal diet without chitosan served as the control group, while the other three groups were fed diets containing 0.2, 0.4 and 0.6 g chitosan/kg diet. We observed significant (P ≤ 0.05) differences among chitosan treatments. Specifically, groups supplemented with chitosan in their diets had significantly (P ≤ 0.05) higher final body weight and weight gain, as well as an improved feed conversion ratio compared to the control group. Rabbits fed chitosan at 0.2 g/kg diet performed the best in growth traits compared to other treatment groups. Rabbits fed chitosan at 0.4 g/kg diet had significantly (P ≤ 0.05) higher levels of total protein in their blood than in the control group. However, levels did not differ significantly from that in the 0.2 group. Likewise, the highest (P ≤ 0.05) levels of glucose and HDL concentrations were detected in rabbits fed chitosan at a 0.2 g/kg diet. All chitosan-supplemented groups (especially at 0.2 g/kg diet) had higher economic efficiency and relative profit levels than the control group. Moreover, morphometric evaluations of the small intestine revealed higher villi number and crypt depth values in chitosan-treated animals compared to the control group. We conclude that using chitosan at levels of 0.2 or 0.4 g/kg diet can effectively improve the body weight gain, feed conversion ratio and economic efficiency, as well as the overall health status of NZW under Egyptian conditions.

Sexual behaviour response, testicular development and semen quality of New Zealand white rabbit bucks as influenced by dietary chitosan.

The present study was conducted to determine the effects of dietary chitosan supplementation on sexual behaviour responses, testicular development, and semen quality traits of New Zealand White (NZW) rabbit bucks. Twenty-four 5-week-old rabbit bucks were used in this experiment. Animals were grouped into four equal experimental groups: the control group was fed only on a basal diet, whereas the other groups were fed the basal diet supplemented with three levels of chitosan at 0.2, 0.4, or 0.6 g/kg, respectively. Also, bucks that received chitosan at 0.2 and 0.4 g/kg had a significantly earlier time of sexual libido (p ≤ .05) and had significantly higher ejaculate volume and sperm concentration than other groups (p ≤ .001). Furthermore, basic and sexual behaviours were significantly improved in bucks fed chitosan at 0.2 and 0.4 g/kg compared with other groups. Therefore, it could be concluded that using chitosan at 0.2 and 0.4 g/kg enhanced sexual behaviour, improved semen quality, and reproductive efficiency in the NZW rabbit bucks.

Topography of pleural epithelial structure enabled by en face isolation and machine learning.

Pleural epithelial adaptations to mechanical stress are relevant to both normal lung function and parenchymal lung diseases. Assessing regional differences in mechanical stress, however, has been complicated by the nonlinear stress-strain properties of the lung and the large displacements with ventilation. Moreover, there is no reliable method of isolating pleural epithelium for structural studies. To define the topographic variation in pleural structure, we developed a method of en face harvest of murine pleural epithelium. Silver-stain was used to highlight cell borders and facilitate imaging with light microscopy. Machine learning and watershed segmentation were used to define the cell area and cell perimeter of the isolated pleural epithelial cells. In the deflated lung at residual volume, the pleural epithelial cells were significantly larger in the apex (624 ± 247 µm2 ) than in basilar regions of the lung (471 ± 119 µm2 ) (p < 0.001). The distortion of apical epithelial cells was consistent with a vertical gradient of pleural pressures. To assess epithelial changes with inflation, the pleura was studied at total lung capacity. The average epithelial cell area increased 57% and the average perimeter increased 27% between residual volume and total lung capacity. The increase in lung volume was less than half the percent change predicted by uniform or isotropic expansion of the lung. We conclude that the structured analysis of pleural epithelial cells complements studies of pulmonary microstructure and provides useful insights into the regional distribution of mechanical stresses in the lung.

Facilitated Transport across Glycocalyceal Barriers in the Chick Chorioallantoic Membrane.

Targeted drug delivery to visceral organs offers the possibility of not only limiting the required dose, but also minimizing drug toxicity; however, there is no reliable method for delivering drugs to the surface of visceral organs. Here, we used six color tracers and the chick chorioallantoic membrane (CAM) model to investigate the use of the heteropolysaccharide pectin to facilitate tracer diffusion across the glycocalyceal charge barrier. The color tracers included brilliant blue, Congo red, crystal violet, indocyanine green, methylene blue, and methyl green. The direct application of the tracers to the CAM surface or embedding tracers into linear-chain nanocellulose fiber films resulted in no significant diffusion into the CAM. In contrast, when the tracers were actively loaded into branched-chain pectin films, there was significant detectable diffusion of the tracers into the CAM. The facilitated diffusion was observed in the three cationic tracers but was limited in the three anionic tracers. Diffusion appeared to be dependent on ionic charge, but independent of tracer size or molecular mass. We conclude that dye-loaded pectin films facilitated the diffusion of color tracers across the glycocalyceal charge barrier and may provide a therapeutic path for drug delivery to the surface of visceral organs.

Kinetics of Pectin Biopolymer Facial Erosion Characterized by Fluorescent Tracer Microfluidics.

Pectin is a plant-derived heteropolysaccharide that has been implicated in drug development, tissue engineering, and visceral organ repair. Pectin demonstrates remarkable biostability in a variety of physiologic environments but is biodegradable in water. To understand the dynamics of pectin biodegradation in basic environments, we developed a microfluidics system that facilitated the quantitative comparison of pectin films exposed to facial erosion. Pectin biodegradation was assessed using fluorescein tracer embedded in pectin, trypan blue quenching of released fluorescence, and highly sensitive microfluorimetry. The microfluidic perfusate, delivered through 6 um-pore synthetic membrane interface, demonstrated nonlinear erosion of the pectin film; 75% of tracer was released in 28 h. The microfluidics system was used to identify potential modifiers of pectin erosion. The polyphenolic compound tannic acid, loaded into citrus pectin films, demonstrated a dose-dependent decrease in pectin erosion. Tannic acid had no detectable impact on the physical properties of citrus pectin including adhesivity and cohesion. In contrast, tannic acid weakened the burst strength and cohesion of pectins derived from soy bean and potato sources. We conclude that facial erosion may explain the biostability of citrus pectin on visceral organ surfaces as well as provide a useful method for identifying modifiers of citrus pectin biodegradation.

Optical and Mechanical Properties of Self-Repairing Pectin Biopolymers.

Pectin's unique physicochemical properties have been linked to a variety of reparative and regenerative processes in nature. To investigate the effect of water on pectin repair, we used a 5 mm stainless-steel uniaxial load to fracture glass phase pectin films. The fractured gel phase films were placed on a 1.5-1.8 mm thick layer of water and incubated for 8 h at room temperature and ambient humidity. There was no immersion or agitation. The repaired pectin film was subsequently assessed for its optical and mechanical properties. Light microscopy demonstrated repair of the detectable fracture area and restoration of the films' optical properties. The burst strength of the repaired film declined to 55% of the original film. However, its resilience was restored to 87% of the original film. Finally, a comparison of the initial and post-repair fracture patterns demonstrated no recurrent fissures in the repaired glass phase films. The water-induced repair of the pectin film was superior to the optical and mechanical properties of the repaired films composed of nanocellulose fibers, sodium hyaluronate, and oxidized cellulose. We conclude that the unique physicochemical properties of pectin facilitate the water-induced self-repair of fractured pectin films.

Bronchopleural fistula due to cavitary pneumonia after SARS-CoV-2 infection treated with open thoracostomy.

Severe coronavirus disease of 2019 (COVID-19) disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection causes substantial parenchymal damage in some patients. There is a paucity of literature describing the surgical management COVID-19 associated bronchopleural fistula after failure of medical therapy. We present the case of a 59-year-old woman with SARS-CoV-2 pneumonia, secondary bacterial pneumonia with bronchopleural fistula and radiographic and clinical evidence of disease refractory to medical therapy. After a course of culture-driven antimicrobial therapy and failure to improve following drainage with tube thoracostomy, she was treated successfully with Clagett open thoracostomy. After resolution of the bronchopleural fistula, the thoracostomy was closed and she was discharged home. In cases of severe COVID-19 complicated by bronchopleural fistula with parenchymal destruction, a tailored approach involving surgical management when indicated can lead to acceptable outcomes without significant morbidity.

Active Loading of Pectin Hydrogels for Targeted Drug Delivery.

Hydrogels provide a promising method for the targeted delivery of protein drugs. Loading the protein drug into the hydrogel free volume can be challenging due to limited quantities of the drug (e.g., growth factor) and complex physicochemical properties of the hydrogel. Here, we investigated both passive and active loading of the heteropolysaccharide hydrogel pectin. Passive loading of glass phase pectin films was evaluated by contact angles and fractional thickness of the pectin films. Four pectin sources demonstrated mean contact angles of 88° with water and 122° with pleural fluid (p < 0.05). Slow kinetics and evaporative losses precluded passive loading. In contrast, active loading of the translucent pectin films was evaluated with the colorimetric tracer methylene blue. Active loading parameters were systematically varied and recorded at 500 points/s. The distribution of the tracer was evaluated by image morphometry. Active loading of the tracer into the pectin films required the optimization of probe velocity, compression force, and contact time. We conclude that active loading using pectin-specific conditions is required for the efficient embedding of low viscosity liquids into pectin hydrogels.