[Association between depressive tendencies and oral diadochokinesis in the older adults in private residential nursing home].

AIM: This study aimed to investigate the relationship between depressive tendencies and oral diadochokinesis (ODK) in 24 older adults living in a private residential nursing home. ODK is an indicator of the oral function. METHODS: Depressive tendencies were assessed using the Geriatric Depression Scale 5, with scores of two or higher indicating probable depression. ODK was measured across four syllable tasks (/pa/, /ta/, /ka/, and /pataka/), which were evaluated using coefficient of variation (CV) values. Low CV values indicate superior performance. Potential confounders, including the cognitive function, sleep status, body mass index, instrumental activities of daily living, and physical function, were controlled. RESULTS: Five participants (20.8%) experienced depression. Individuals with depressive tendencies demonstrated significantly poor ODK performance (higher CV) in the /ta/ task and a marginally significant difference in the /ka/ task. No significant differences were observed between /pa/ and /pataka/. CONCLUSIONS: These findings suggest a link between depressive tendencies and reduced proficiency in specific ODK tasks among older nursing home residents. This finding implies that a decline in the oral function in articulating /ta/ and /ka/ syllables may precede other common depressive symptoms. Furthermore, depression monitoring could be a valuable tool for early detection of the oral function in this population, enabling timely interventions.

Hypochlorous acid water prevents postoperative intrauterine infection after microwave endometrial ablation.

AIM: We investigated the effects of uterine cavity lavage using hypochlorous acid water (HClO) on preventing postoperative intrauterine infection after microwave endometrial ablation (MEA). METHODS: Four hundred and eleven consecutive patients were enrolled in this study. The patients were divided into two groups: 214 cases in 2014-2016 with disinfection using povidone-iodine antiseptic solution alone (group A) and 197 cases in 2017-2019 with additional intrauterine douche using HClO (group B). HClO was used at a concentration of 200 ppm of residual chlorine. One gram of Ceftriaxone Sodium Hydrate was administered by drip infusion during MEA in both groups. Oral antibiotics were administered after MEA only in group A but not in group B. RESULTS: Mean patient age (mean ± SD; years old) was 44.5 ± 4.6 in group A and 44.8 ± 5.4 in group B, and mean operation time (min) was 30.4 ± 19.1 in group A and 34.4 ± 22.6 in group B, respectively. Neither were significantly different between groups. The combined ablation techniques i.e. transcervical microwave myolysis and transcervical microwave adenomyolysis did not increase frequency of infection. Postoperative intrauterine infection cases in group B (8 cases) were significantly lower than those in group A (28 cases) (Chi-square test, P = 0.001). Hysterectomy was performed in three severe intrauterine infection cases in group A, but no cases of severe intrauterine infection was found in group B. No adverse effect of HClO was seen. CONCLUSION: Intrauterine douche using HClO decreases postoperative intrauterine infection after MEA.

Why do myofibroblasts preferentially accumulate on the convex surface of the remodeling lung after pneumonectomy?

Myofibroblasts preferentially accumulate on the convex and not on the concave surfaces of the murine cardiac lobe during lung remodeling after pneumonectomy. This clear difference in function due to the organ shape is most likely mediated by the various mechanical forces generated on the lung's surface. For breathing, the lobe cyclically change its configuration. The cyclic deformation requires energy, depending on the local configuration of the lobe (e.g., convex vs. concave). Considering mechanical contributions to the internal energy of the system and according to the second law of thermodynamics, the system seeks the lowest energy state for equilibrium. Although additional energy for remodeling is required, the system chooses such remodeling sites that minimize the total energy of the new equilibrium state. To test this idea, an idealized, concave-convex configuration of the lobe is assumed. The lobe is made of two homogeneous and isotropic materials of different mechanical properties, the bulk parenchyma and the pleura, a thin, mesothelial cell layer surrounding it. While the whole system cyclically changes shape during breathing, we calculated the amount of mechanical energy per unit volume at the parenchyma-pleural interface where, we believe, myofibroblasts preferentially accumulate. Comparison between convex and concave surfaces indicates that convex surfaces store a lower amount of mechanical energy than the concave ones. We also show that any additional energy for remodeling is preferably done at the convex surface where the lowest new energy equilibrium state is achieved.

Submyometrial vasopressin injection before microwave ablation of vascular-rich submucosal myomas: a preliminary case study.

Purpose: Vascular-rich myomas are resistant to treatment involving transcervical microwave myolysis. To overcome cooling by blood perfusion, we injected dilute vasopressin solution into the space between the myometrium and the surface of the vascular-rich myomas. Material and Methods: Seven outpatients [age (mean ± SD age), 44.9 ± 3.9 years] with a single symptomatic vascular-rich submucosal myoma measuring 4.2-9.2 cm (6.5 ± 2.5 cm) underwent transcervical microwave myolysis and microwave endometrial ablation. Before microwave irradiation, dilute vasopressin solution was injected into the space between the myometrium and the surface of the vascular-rich myoma. We assessed the changes in the volumes of the vascular-rich myomas and blood hemoglobin levels before and 3 and 6 months after treatment. In addition, improvements in menorrhagia and satisfaction after the operation were assessed using visual analog scales. Results: Submyometrial injection of dilute vasopressin effectively reduced the abundant blood flow. The vascular-rich myomas were necrotized and shrank significantly by 69.0% at 3 months and 72.4% at 6 months after the operation (p < .05). Blood hemoglobin levels significantly increased at 3 months (p < .01). In addition, the visual analog scale results indicated that menorrhagia improved subjectively and the patients were satisfied with the results of the operation. Conclusions: Vasopressin injection before transcervical microwave myolysis leads to extended necrosis of vascular-rich submucosal myomas.

Deformation-induced transitional myofibroblasts contribute to compensatory lung growth.

In many mammals, including humans, removal of one lung (pneumonectomy) results in the compensatory growth of the remaining lung. Compensatory growth involves not only an increase in lung size, but also an increase in the number of alveoli in the peripheral lung; however, the process of compensatory neoalveolarization remains poorly understood. Here, we show that the expression of α-smooth muscle actin (SMA)-a cytoplasmic protein characteristic of myofibroblasts-is induced in the pleura following pneumonectomy. SMA induction appears to be dependent on pleural deformation (stretch) as induction is prevented by plombage or phrenic nerve transection (P < 0.001). Within 3 days of pneumonectomy, the frequency of SMA+ cells in subpleural alveolar ducts was significantly increased (P < 0.01). To determine the functional activity of these SMA+ cells, we isolated regenerating alveolar ducts by laser microdissection and analyzed individual cells using microfluidic single-cell quantitative PCR. Single cells expressing the SMA (Acta2) gene demonstrated significantly greater transcriptional activity than endothelial cells or other discrete cell populations in the alveolar duct (P < 0.05). The transcriptional activity of the Acta2+ cells, including expression of TGF signaling as well as repair-related genes, suggests that these myofibroblast-like cells contribute to compensatory lung growth.

Alveolar septal patterning during compensatory lung growth: Part II the effect of parenchymal pressure gradients.

In most mammals, compensatory lung growth occurs after the removal of one lung (pneumonectomy). Although the mechanism of alveolar growth is unknown, the patterning of complex alveolar geometry over organ-sized length scales is a central question in regenerative lung biology. Because shear forces appear capable of signaling the differentiation of important cells involved in neoalveolarization (fibroblasts and myofibroblasts), interstitial fluid mechanics provide a potential mechanism for the patterning of alveolar growth. The movement of interstitial fluid is created by two basic mechanisms: 1) the non-uniform motion of the boundary walls, and 2) parenchymal pressure gradients external to the interstitial fluid. In a previous study (Haber et al., Journal of Theoretical Biology 400: 118-128, 2016), we investigated the effects of non-uniform stretching of the primary septum (associated with its heterogeneous mechanical properties) during breathing on generating non-uniform Stokes flow in the interstitial space. In the present study, we analyzed the effect of parenchymal pressure gradients on interstitial flow. Dependent upon lung microarchitecture and physiologic conditions, parenchymal pressure gradients had a significant effect on the shear stress distribution in the interstitial space of primary septa. A dimensionless parameter δ described the ratio between the effects of a pressure gradient and the influence of non-uniform primary septal wall motion. Assuming that secondary septa are formed where shear stresses were the largest, it is shown that the geometry of the newly generated secondary septa was governed by the value of δ. For δ smaller than 0.26, the alveolus size was halved while for higher values its original size was unaltered. We conclude that the movement of interstitial fluid, governed by parenchymal pressure gradients and non-uniform primary septa wall motion, provides a plausible mechanism for the patterning of alveolar growth.

Structural and functional evidence for the scaffolding effect of alveolar blood vessels.

A contribution of pulmonary blood distension to alveolar opening was first proposed more than 100 years ago. To investigate the contribution of blood distension to lung mechanics, we studied control mice (normal perfusion), mice after exsanguination (absent perfusion) and mice after varying degrees of parenchymal resection (supra-normal perfusion). On inflation, mean tracheal pressures were higher in the bloodless mouse (4.0 ± 2.5 cm H2O); however, there was minimal difference between conditions on deflation (0.7 ± 0.9 cm H2O). To separate the peripheral and central mechanical effects of blood volume, multi-frequency lung impedance data was fitted to the constant-phase model. The presence or absence of blood had no effect on central airway resistance (p > .05). In contrast, measures of tissue damping (G), tissue elastance (H) and hysteresivity (η) demonstrated a significant increase in bloodless mice relative to control mice (p < .001). After varying amount of surgical resection and associated supra-normal perfusion of the remaining lung, there was an increase in G and H. Although the absolute difference in G and H increased with the amount of parenchymal resection, the proportional contribution of blood was identical in all conditions. The presence of blood in the pulmonary vasculature resulted in a constant 64 ± 5% reduction in tissue damping (G) and a 55 ± 4% reduction in tissue elastance (H). This nearly-constant contribution of blood to lung hysteresivity was only reduced by positive end-expiratory pressure (PEEP). To identify a distinct structural subset of vessels in the lung potentially contributing to these observations, vascular casting and scanning electron microscopy of the lung demonstrated morphologically distinct vascular rings at the alveolar opening. Our results suggest that intravascular blood distension, likely attributable to a subset of vessels in the alveolar entrance ring, contributes a measurable scaffolding effect to the functional recruitment of the peripheral lung.

Interstitial fluid flow of alveolar primary septa after pneumonectomy.

Neoalveolation is known to occur in the remaining lung after pneumonectomy. While compensatory lung growth is a complex process, stretching of the lung tissue appears to be crucial for tissue remodeling. Even a minute shear stress exerted on fibroblasts in the interstitial space is known to trigger cell differentiation into myofibroblast that are essential to building new tissues. We hypothesize that the non-uniform motion of the primary septa due to their heterogeneous mechanical properties under tidal breathing induces a spatially unique interstitial flow and shear stress distribution in the interstitial space. This may in turn trigger pulmonary fibroblast differentiation and neoalveolation. In this study, we developed a theoretical basis for how cyclic motion of the primary septal walls with heterogeneous mechanical properties affects the interstitial flow and shear stress distribution. The velocity field of the interstitial flow was expressed by a Fourier (complex) series and its leading term was considered to induce the basic structure of stress distribution as long as the dominant length scale of heterogeneity is the size of collapsed alveoli. We conclude that the alteration of mechanical properties of the primary septa caused by pneumonectomy can develop a new interstitial flow field, which alters the shear stress distribution. This may trigger the differentiation of resident fibroblasts, which may in turn induce spatially unique neoalveolation in the remaining lung. Our example illustrates that the initial forming of new alveoli about half the size of the original ones.

Remodeling of alveolar septa after murine pneumonectomy.

In most mammals, removing one lung (pneumonectomy) results in the compensatory growth of the remaining lung. In mice, stereological observations have demonstrated an increase in the number of mature alveoli; however, anatomic evidence of the early phases of alveolar growth has remained elusive. To identify changes in the lung microstructure associated with neoalveolarization, we used tissue histology, electron microscopy, and synchrotron imaging to examine the configuration of the alveolar duct after murine pneumonectomy. Systematic histological examination of the cardiac lobe demonstrated no change in the relative frequency of dihedral angle components (Ends, Bends, and Junctions) (P > 0.05), but a significant decrease in the length of a subset of septal ends ("E"). Septal retraction, observed in 20-30% of the alveolar ducts, was maximal on day 3 after pneumonectomy (P < 0.01) and returned to baseline levels within 3 wk. Consistent with septal retraction, the postpneumonectomy alveolar duct diameter ratio (Dout:Din) was significantly lower 3 days after pneumonectomy compared to all controls except for the detergent-treated lung (P < 0.001). To identify clumped capillaries predicted by septal retraction, vascular casting, analyzed by both scanning electron microscopy and synchrotron imaging, demonstrated matted capillaries that were most prominent 3 days after pneumonectomy. Numerical simulations suggested that septal retraction could reflect increased surface tension within the alveolar duct, resulting in a new equilibrium at a higher total energy and lower surface area. The spatial and temporal association of these microstructural changes with postpneumonectomy lung growth suggests that these changes represent an early phase of alveolar duct remodeling.

Evidence for adult lung growth in humans.

A 33-year-old woman underwent a right-sided pneumonectomy in 1995 for treatment of a lung adenocarcinoma. As expected, there was an abrupt decrease in her vital capacity, but unexpectedly, it increased during the subsequent 15 years. Serial computed tomographic (CT) scans showed progressive enlargement of the remaining left lung and an increase in tissue density. Magnetic resonance imaging (MRI) with the use of hyperpolarized helium-3 gas showed overall acinar-airway dimensions that were consistent with an increase in the alveolar number rather than the enlargement of existing alveoli, but the alveoli in the growing lung were shallower than in normal lungs. This study provides evidence that new lung growth can occur in an adult human.

Outpatient transcervical microwave myolysis assisted by transabdominal ultrasonic guidance for menorrhagia caused by submucosal myomas.

PURPOSE: The aim of this paper was to evaluate the effectiveness in day clinics of microwave endometrial ablation (MEA) on transcervical microwave myolysis for patients with menorrhagia caused by submucosal myomas. MATERIALS AND METHODS: Thirty-five outpatients (average age 44.8 ± 5.2 years (mean ± SD), range 34-58) with a single submucosal myoma that was 4-7 cm (5.5 ± 2.1 cm) in size underwent MEA with transcervical microwave myolysis using a specifically developed transabdominal ultrasound probe attachment for transcervical puncture. Primary outcomes were the changes in the blood haemoglobin level and the volume of myoma before and after the treatment. Secondary outcomes were the improvement in menorrhagia and satisfaction after the operation, assessed by visual analogue scale (VAS). RESULTS: The mean operation time was 27.9 ± 13.6 min. The myomas had shrunk by 56.2% at 3 months and 73.8% at ≥6 months after the operation. Blood haemoglobin levels had increased significantly at 3 months (10.2 ± 2.0 vs. 12.7 ± 1.2, p < 0.001). The average VAS assessment of menstrual bleeding had decreased to 1.7 ± 1.7 at 3 months after the operation (preoperative VAS = 10). The average VAS score for feelings of satisfaction 3 months after the operation was 9.8 ± 0.5 (full score = 10). CONCLUSIONS: MEA with transcervical microwave myolysis is a feasible and effective procedure in a day surgery clinic for menorrhagia caused by submucosal myomas. The procedure may be an alternative to hysterectomy for menorrhagia caused by submucosal myomas in women during the perimenopausal period.

Nanoparticle delivery in infant lungs.

The lung surface is an ideal pathway to the bloodstream for nanoparticle-based drug delivery. Thus far, research has focused on the lungs of adults, and little is known about nanoparticle behavior in the immature lungs of infants. Here, using nonlinear dynamical systems analysis and in vivo experimentation in developing animals, we show that nanoparticle deposition in postnatally developing lungs peaks at the end of bulk alveolation. This finding suggests a unique paradigm, consistent with the emerging theory that as alveoli form through secondary septation, alveolar flow becomes chaotic and chaotic mixing kicks in, significantly enhancing particle deposition. This finding has significant implications for the application of nanoparticle-based inhalation therapeutics in young children with immature lungs from birth to 2 y of age.

Sprouting and intussusceptive angiogenesis in postpneumonectomy lung growth: mechanisms of alveolar neovascularization.

In most rodents and some other mammals, the removal of one lung results in compensatory growth associated with dramatic angiogenesis and complete restoration of lung capacity. One pivotal mechanism in neoalveolarization is neovascularization, because without angiogenesis new alveoli can not be formed. The aim of this study is to image and analyze three-dimensionally the different patterns of neovascularization seen following pneumonectomy in mice on a sub-micron-scale. C57/BL6 mice underwent a left-sided pneumonectomy. Lungs were harvested at various timepoints after pneumonectomy. Volume analysis by microCT revealed a striking increase of 143 percent in the cardiac lobe 14 days after pneumonectomy. Analysis of microvascular corrosion casting demonstrated spatially heterogenous vascular densitities which were in line with the perivascular and subpleural compensatory growth pattern observed in anti-PCNA-stained lung sections. Within these regions an expansion of the vascular plexus with increased pillar formations and sprouting angiogenesis, originating both from pre-existing bronchial and pulmonary vessels was observed. Also, type II pneumocytes and alveolar macrophages were seen to participate actively in alveolar neo-angiogenesis after pneumonectomy. 3D-visualizations obtained by high-resolution synchrotron radiation X-ray tomographic microscopy showed the appearance of double-layered vessels and bud-like alveolar baskets as have already been described in normal lung development. Scanning electron microscopy data of microvascular architecture also revealed a replication of perialveolar vessel networks through septum formation as already seen in developmental alveolarization. In addition, the appearance of pillar formations and duplications on alveolar entrance ring vessels in mature alveoli are indicative of vascular remodeling. These findings indicate that sprouting and intussusceptive angiogenesis are pivotal mechanisms in adult lung alveolarization after pneumonectomy. Various forms of developmental neoalveolarization may also be considered to contribute in compensatory lung regeneration.

Stretch-induced intussuceptive and sprouting angiogenesis in the chick chorioallantoic membrane.

Vascular systems grow and remodel in response to not only metabolic needs, but also mechanical influences as well. Here, we investigated the influence of tissue-level mechanical forces on the patterning and structure of the chick chorioallantoic membrane (CAM) microcirculation. A dipole stretch field was applied to the CAM using custom computer-controlled servomotors. The topography of the stretch field was mapped using finite element models. After 3days of stretch, Sholl analysis of the CAM demonstrated a 7-fold increase in conducting vessel intersections within the stretch field (p<0.01). The morphometric analysis of intravital microscopy and scanning electron microscopy (SEM) images demonstrated that the increase vessel density was a result of an increase in interbranch distance (p<0.01) and a decrease in bifurcation angles (p<0.01); there was no significant increase in conducting vessel number (p>0.05). In contrast, corrosion casting and SEM of the stretch field capillary meshwork demonstrated intense sprouting and intussusceptive angiogenesis. Both planar surface area (p<0.05) and pillar density (p<0.01) were significantly increased relative to control regions of the CAM. We conclude that a uniaxial stretch field stimulates the axial growth and realignment of conducting vessels as well as intussusceptive and sprouting angiogenesis within the gas exchange capillaries of the ex ovo CAM.

Randomised controlled trial of the effects of L-ornithine on stress markers and sleep quality in healthy workers.

BACKGROUND: L-ornithine is a non-essential, non-protein amino acid. Although L-ornithine is contained in various foods, the amount is usually small.Recently, studies have shown that orally administered L-ornithine reduced the stress response in animals.From these findings, we speculated that L-ornithine may play a role in the relieve of stress and improve sleep and fatigue symptoms in humans. Through a randomised, double-blind, placebo-controlled clinical study, we asked if L-ornithine could be beneficial to stress and sleep in healthy workers. METHOD: Fifty-two apparently healthy Japanese adults who had previously felt slight stress as well as fatigue were recruited to be study participants and were randomly divided into either the L-ornithine (400 mg/day) or placebo group. They orally consumed the respective test substance every day for 8 weeks. Serum was collected for the assessment of cortisol and dehydroepiandrosterone-sulphate (DHEA-S). Perceived mood and quality of sleep were measured by the Profile of Mood States (POMS), Athens Insomnia Scale (AIS), and Ogri-Shirakawa-Azumi sleep inventory MA version (OSA-MA). RESULTS: Serum cortisol levels and the cortisol/DHEA-S ratio were significantly decreased in the L-ornithine group in comparison with the placebo group. Also, anger was reduced and perceived sleep quality was improved in the L-ornithine group. CONCLUSION: L-ornithine supplementation has the potential to relieve stress and improve sleep quality related to fatigue, both objectively and subjectively.

Intussusceptive remodeling of vascular branch angles in chemically-induced murine colitis.

Intussusceptive angiogenesis is a developmental process linked to both blood vessel replication and remodeling in development. To investigate the prediction that the process of intussusceptive angiogenesis is associated with vessel angle remodeling in adult mice, we systematically evaluated corrosion casts of the mucosal plexus in mice with trinitrobenzesulfonic acid (TNBS)-induced and dextran sodium sulfate (DSS)-induced colitis. The mice demonstrated a significant decrease in vessel angles in both TNBS-induced and DSS-induced colitis within 4 weeks of the onset of colitis (p<.001). Corrosion casts 28-30 days after DSS treatment were studied for a variety of detailed morphometric changes. The vessel diameter and interbranch distance were significantly increased in the descending colon (p<.05). Also consistent with vessel growth, intervascular distance was decreased in the descending colon (p<.05). In contrast, no statistically significant morphometric changes were noted in the ascending colon. The morphometry of the corrosion casts also demonstrated 1) a similar orientation of the remodeled angles within the XY coordinate plane of the mucosal plexus, and 2) alternating periodicity of remodeled and unremodeled vessel angles. We conclude that inflammation-associated intussusceptive angiogenesis in adult mice is associated with vessel angle remodeling. Further, the morphometry of the vessel angles suggests the influence of blood flow on the location and orientation of remodeled vessels.

A mesoscopic bridging scale method for fluids and coupling dissipative particle dynamics with continuum finite element method.

A multiscale procedure to couple a mesoscale discrete particle model and a macroscale continuum model of incompressible fluid flow is proposed in this study. We call this procedure the mesoscopic bridging scale (MBS) method since it is developed on the basis of the bridging scale method for coupling molecular dynamics and finite element models [G.J. Wagner, W.K. Liu, Coupling of atomistic and continuum simulations using a bridging scale decomposition, J. Comput. Phys. 190 (2003) 249-274]. We derive the governing equations of the MBS method and show that the differential equations of motion of the mesoscale discrete particle model and finite element (FE) model are only coupled through the force terms. Based on this coupling, we express the finite element equations which rely on the Navier-Stokes and continuity equations, in a way that the internal nodal FE forces are evaluated using viscous stresses from the mesoscale model. The dissipative particle dynamics (DPD) method for the discrete particle mesoscale model is employed. The entire fluid domain is divided into a local domain and a global domain. Fluid flow in the local domain is modeled with both DPD and FE method, while fluid flow in the global domain is modeled by the FE method only. The MBS method is suitable for modeling complex (colloidal) fluid flows, where continuum methods are sufficiently accurate only in the large fluid domain, while small, local regions of particular interest require detailed modeling by mesoscopic discrete particles. Solved examples - simple Poiseuille and driven cavity flows illustrate the applicability of the proposed MBS method.

Application of Machine Learning for Segmentation of the Pulmonary Acinus Imaged by Synchrotron X-Ray Tomography.

Background: To assess the effectiveness of inhalation therapy, it is important to evaluate the lungs' structure; thus, visualization of the entire lungs at the level of the alveoli is necessary. To achieve this goal, the applied visualization technique must satisfy the following two conditions simultaneously: (1) it has to obtain images of the entire lungs, since one part of the lungs is influenced by the other parts, and (2) the images have to capture the detailed structure of the alveolus/acinus in which gas exchange occurs. However, current visualization techniques do not fulfill these two conditions simultaneously. Segmentation is a process in which each pixel of the obtained high-resolution images is simplified (i.e., the representation of an image is changed by categorizing and modifying each pixel) so that we can perform three-dimensional volume rendering. One of the bottlenecks of current approaches is that the accuracy of the segmentation of each image has to be evaluated on the outcome of the process (mainly by an expert). It is a formidable task to evaluate the astronomically large numbers of images that would be required to resolve the entire lungs in high resolution. Methods: To overcome this challenge, we propose a new approach based on machine learning (ML) techniques for the validation step. Results: We demonstrate the accuracy of the segmentation process itself by comparison with previously validated images. In this ML approach, to achieve a reasonable accuracy, millions/billions of parameters used for segmentation have to be optimized. This computationally demanding new approach is achievable only due to recent dramatic increases in computation power. Conclusion: The objective of this article is to explain the advantages of ML over the classical approach for acinar imaging.

CD34+ progenitor to endothelial cell transition in post-pneumonectomy angiogenesis.

In many species, pneumonectomy triggers compensatory lung growth that results in an increase not only in lung volume, but also in alveolar number. Whether the associated alveolar angiogenesis involves the contribution of blood-borne progenitor cells is unknown. To identify and characterize blood-borne progenitor cells contributing to lung growth after pneumonectomy in mice, we studied wild-type and wild-type/green fluorescence protein (GFP) parabiotic mice after left pneumonectomy. Within 21 days of pneumonectomy, a 3.2-fold increase occurred in the number of lung endothelial cells. This increase in total endothelial cells was temporally associated with a 7.3-fold increase in the number of CD34(+) endothelial cells. Seventeen percent of the CD34(+) endothelial cells were actively proliferating, compared with only 4.2% of CD34(-) endothelial cells. Using wild-type/GFP parabiotic mice, we demonstrated that 73.4% of CD34(+) cells were derived from the peripheral blood. Furthermore, lectin perfusion studies demonstrated that CD34(+) cells derived from peripheral blood were almost uniformly incorporated into the lung vasculature. Finally, CD34(+) endothelial cells demonstrated a similar profile, but had enhanced transcriptional activity relative to CD34(-) endothelial cells. We conclude that blood-borne CD34(+) endothelial progenitor cells, characterized by active cell division and an amplified transcriptional signature, transition into resident endothelial cells during compensatory lung growth.

Alveolar macrophage dynamics in murine lung regeneration.

In most mammalian species, the removal of one lung results in dramatic compensatory growth of the remaining lung. To investigate the contribution of alveolar macrophages (AMs) to murine post-pneumonectomy lung growth, we studied bronchoalveolar lavage (BAL)-derived AM on 3, 7, 14 and 21 days after left pneumonectomy. BAL demonstrated a 3.0-fold increase in AM (CD45(+), CD11b(-), CD11c(+), F4/80(+), Gr-1(-)) by 14 days after pneumonectomy. Cell cycle flow cytometry of the BAL-derived cells demonstrated an increase in S + G2 phase cells on days 3 (11.3 ± 2.7%) and 7 (12.1 ± 1.8%) after pneumonectomy. Correspondingly, AM demonstrated increased expression of VEGFR1 and MHC class II between days 3 and 14 after pneumonectomy. To investigate the potential contribution of peripheral blood cells to this AM population, parabiotic mice (wild-type/GFP) underwent left pneumonectomy. Analysis of GFP(+) cells in the post-pneumonectomy lung demonstrated that by day 14, less than 1% of the AM population were derived from the peripheral blood. Finally, AM gene transcription demonstrated a significant shift from decreased transcription of angiogenesis-related genes on day 3 to increased transcription on day 7 after pneumonectomy. The increased number of locally proliferating AM, combined with their growth-related gene transcription, suggests that AM actively participate in compensatory lung growth.