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  • 1
    Online Resource
    Online Resource
    Cell-Demanded Liberation of VEGF 121 From Fibrin Implants Induces Local and Controlled Blood Vessel Growth
    Ovid Technologies (Wolters Kluwer Health) ; 2004
    In:  Circulation Research Vol. 94, No. 8 ( 2004-04-30), p. 1124-1132
    Details
    In: Circulation Research, Ovid Technologies (Wolters Kluwer Health), Vol. 94, No. 8 ( 2004-04-30), p. 1124-1132
    Abstract: Although vascular endothelial growth factor (VEGF) has been described as a potent angiogenic stimulus, its application in therapy remains difficult: blood vessels formed by exposure to VEGF tend to be malformed and leaky. In nature, the principal form of VEGF possesses a binding site for ECM components that maintain it in the immobilized state until released by local cellular enzymatic activity. In this study, we present an engineered variant form of VEGF, α 2 PI 1–8 -VEGF 121 , that mimics this concept of matrix-binding and cell-mediated release by local cell-associated enzymatic activity, working in the surgically-relevant biological matrix fibrin. We show that matrix-conjugated α 2 PI 1–8 -VEGF 121 is protected from clearance, contrary to native VEGF 121 mixed into fibrin, which was completely released as a passive diffusive burst. Grafting studies on the embryonic chicken chorioallantoic membrane (CAM) and in adult mice were performed to assess and compare the quantity and quality of neovasculature induced in response to fibrin implants formulated with matrix-bound α 2 PI 1–8 -VEGF 121 or native diffusible VEGF 121 . Our CAM measurements demonstrated that cell-demanded release of α 2 PI 1–8 -VEGF 121 increases the formation of new arterial and venous branches, whereas exposure to passively released wild-type VEGF 121 primarily induced chaotic changes within the capillary plexus. Specifically, our analyses at several levels, from endothelial cell morphology and endothelial interactions with periendothelial cells, to vessel branching and network organization, revealed that α 2 PI 1–8 -VEGF 121 induces vessel formation more potently than native VEGF 121 and that those vessels possess more normal morphologies at the light microscopic and ultrastructural level. Permeability studies in mice validated that vessels induced by α 2 PI 1–8 -VEGF 121 do not leak. In conclusion, cell-demanded release of engineered VEGF 121 from fibrin implants may present a therapeutically safe and practical modality to induce local angiogenesis.
    Type of Medium: Online Resource
    ISSN: 0009-7330 , 1524-4571
    URL: Article
    DOI: 10.1161/01.RES.0000126411.29641.08
    RVK:
    XA 10000
    Language: English
    Publisher: Ovid Technologies (Wolters Kluwer Health)
    Publication Date: 2004
    detail.hit.zdb_id: 1467838-X
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  • 2
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    Online Resource
    Rat lungs show a biphasic formation of new alveoli during postnatal development
    American Physiological Society ; 2014
    In:  Journal of Applied Physiology Vol. 117, No. 1 ( 2014-07-01), p. 89-95
    Details
    In: Journal of Applied Physiology, American Physiological Society, Vol. 117, No. 1 ( 2014-07-01), p. 89-95
    Abstract: Roughly 90% of the gas-exchange surface is formed by alveolarization of the lungs. To the best of our knowledge, the formation of new alveoli has been followed in rats only by means of morphological description or interpretation of semiquantitative data until now. Therefore, we estimated the number of alveoli in rat lungs between postnatal days 4 and 60 by unambiguously counting the alveolar openings. We observed a bulk formation of new alveoli between days 4 and 21 (17.4 times increase from 0.8 to 14.3 millions) and a second phase of continued alveolarization between days 21 and 60 (1.3 times increase to 19.3 million). The (number weighted) mean volume of the alveoli decreases during the phase of bulk alveolarization from ∼593,000 μm 3 at day 4 to ∼141,000 μm 3 at day 21, but increases again to ∼298,000 μm 3 at day 60. We conclude that the “bulk alveolarization” correlates with the mechanism of classical alveolarization (alveolarization before the microvascular maturation is completed) and that the “continued alveolarization” follows three proposed mechanisms of late alveolarization (alveolarization after microvascular maturation). The biphasic pattern is more evident for the increase in alveolar number than for the formation of new alveolar septa (estimated as the length of the free septal edge). Furthermore, a striking negative correlation between the estimated alveolar size and published data on retention of nanoparticles was detected.
    Type of Medium: Online Resource
    ISSN: 8750-7587 , 1522-1601
    URL: Article
    DOI: 10.1152/japplphysiol.01355.2013
    RVK:
    WA 15000
    RVK:
    XA 52094
    Language: English
    Publisher: American Physiological Society
    Publication Date: 2014
    detail.hit.zdb_id: 1404365-8
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    SSG: 31
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  • 3
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    Online Resource
    EpCAM + CD73 + mark epithelial progenitor cells in postnatal human lung and are associated with pathogenesis of pulmonary disease including lung adenocarcinoma
    American Physiological Society ; 2020
    In:  American Journal of Physiology-Lung Cellular and Molecular Physiology Vol. 319, No. 5 ( 2020-11-01), p. L794-L809
    Details
    In: American Journal of Physiology-Lung Cellular and Molecular Physiology, American Physiological Society, Vol. 319, No. 5 ( 2020-11-01), p. L794-L809
    Abstract: Lung injury in mice induces mobilization of discrete subsets of epithelial progenitor cells to promote new airway and alveolar structures. However, whether similar cell types exist in human lung remains unresolved. Using flow cytometry, we identified a distinct cluster of cells expressing the epithelial cell adhesion molecule (EpCAM), a cell surface marker expressed on epithelial progenitor cells, enriched in the ecto-5′-nucleotidase CD73 in unaffected postnatal human lungs resected from pediatric patients with congenital lung lesions. Within the EpCAM + CD73 + population, a small subset coexpresses integrin β4 and HTII-280. This population remained stable with age. Spatially, EpCAM + CD73 + cells were positioned along the basal membrane of respiratory epithelium and alveolus next to CD73 + cells lacking EpCAM. Expanded EpCAM + CD73 + cells give rise to a pseudostratified epithelium in a two-dimensional air–liquid interface or a clonal three-dimensional organoid assay. Organoids generated under alveolar differentiation conditions were cystic-like and lacked robust alveolar mature cell types. Compared with unaffected postnatal lung, congenital lung lesions were marked by clusters of EpCAM + CD73 + cells in airway and cystic distal lung structures lined by simple epithelium composed of EpCAM + SCGB1A1 + cells and hyperplastic EpCAM + proSPC + cells. In non-small-cell lung cancer (NSCLC), there was a marked increase in EpCAM + CD73 + tumor cells enriched in inhibitory immune checkpoint molecules CD47 and programmed death-ligand 1 (PD-L1), which was associated with poor survival in lung adenocarcinoma (LUAD). In conclusion, EpCAM + CD73 + cells are rare novel epithelial progenitor cells in the human lung. Importantly, reemergence of CD73 in lung adenocarcinoma enriched in negative immune checkpoint molecules may serve as a novel therapeutic target.
    Type of Medium: Online Resource
    ISSN: 1040-0605 , 1522-1504
    URL: Article
    DOI: 10.1152/ajplung.00279.2019
    Language: English
    Publisher: American Physiological Society
    Publication Date: 2020
    detail.hit.zdb_id: 1477300-4
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  • 4
    Online Resource
    Online Resource
    The influence of age on valve disease in patients with varicose veins analysed by transmission electron microscopy and stereology
    Hogrefe Publishing Group ; 2018
    In:  Vasa Vol. 47, No. 5 ( 2018-08-01), p. 409-416
    Details
    In: Vasa, Hogrefe Publishing Group, Vol. 47, No. 5 ( 2018-08-01), p. 409-416
    Abstract: Abstract. Background: The aim of this study was to investigate the influence of age on the ultrastructure of venous valve morphology in patients with C2 classified chronic venous disorders according to the CEAP classification. Patients and methods: The study population consisted of 16 consecutive patients with varicose veins (C2). The mean age was 49.8 years (30–66). The (pre-) terminal valve including the vessel wall was harvested within the proximal 2 centimetres of the great saphenous vein. The mean thickness (volume-to-surface ratio = V/S ratio) of elastin, collagen, endothelium and of the entire valve was determined. A blinded morphologist performed the examination by transmission electron microscopy and stereology. Analyses by Pearson’s product moment correlation, Kendall’s tau and Spearman’s rank correlation were performed to investigate whether there is a correlation between age and the ultrastructural morphology. Results: Stereological analysis of the valves demonstrated a mean V/S ratio (signifying a thickness estimation) for elastin of 0.87 μm 3 /μm 2 , for collagen of 18.0 μm 3 /μm 2 , for endothelium of 0.65 μm 3 /μm 2 , and for the entire valve of 25.2 μm³/μm². Statistical analyses showed no statistically significant correlation between age and the ultrastructural morphology in this patient group. Conclusions: The ultrastructural morphology of the venous valves in chronic venous disorders may not depend on age in patients presenting with C2 disease. This conclusion may or may not apply to all C classes as we investigated a homogenous group of patients with C2 limbs.
    Type of Medium: Online Resource
    ISSN: 0301-1526 , 1664-2872
    URL: Article
    DOI: 10.1024/0301-1526/a000714
    Language: English
    Publisher: Hogrefe Publishing Group
    Publication Date: 2018
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  • 5
    Online Resource
    Online Resource
    Capillary ultrastructure and mitochondrial volume density in skeletal muscle in relation to reduced exercise capacity of patients with intermittent claudication
    American Physiological Society ; 2016
    In:  American Journal of Physiology-Regulatory, Integrative and Comparative Physiology Vol. 310, No. 10 ( 2016-05-15), p. R943-R951
    Details
    In: American Journal of Physiology-Regulatory, Integrative and Comparative Physiology, American Physiological Society, Vol. 310, No. 10 ( 2016-05-15), p. R943-R951
    Abstract: Intermittent claudication (IC) is the most commonly reported symptom of peripheral arterial disease (PAD). Impaired limb blood flow is a major casual factor of lower exercise tolerance in PAD but cannot entirely explain it. We hypothesized that IC is associated with structural changes of the capillary-mitochondria interface that could contribute to the reduction of exercise tolerance in IC patients. Capillary and mitochondrial morphometry were performed after light and transmission electron microscopy using vastus lateralis muscle biopsies of 14 IC patients and 10 age-matched controls, and peak power output (PPO) was determined for all participants using an incremental single-leg knee-extension protocol. Capillary density was lower (411 ± 90 mm −2 vs. 506 ± 95 mm −2 ; P ≤ 0.05) in the biopsies of the IC patients than in those of the controls. The basement membrane (BM) around capillaries was thicker (543 ± 82 nm vs. 423 ± 97 nm; P ≤ 0.01) and the volume density of mitochondria was lower (3.51 ± 0.56% vs. 4.60 ± 0.74%; P ≤ 0.01) in the IC patients than the controls. In the IC patients, a higher proportion of capillaries appeared with collapsed slit-like lumen and/or swollen endothelium. PPO was lower (18.5 ± 9.9 W vs. 33.5 ± 9.4 W; P ≤ 0.01) in the IC patients than the controls. We suggest that several structural alterations in skeletal muscle, either collectively or separately, contribute to the reduction of exercise tolerance in IC patients.
    Type of Medium: Online Resource
    ISSN: 0363-6119 , 1522-1490
    URL: Article
    DOI: 10.1152/ajpregu.00480.2015
    Language: English
    Publisher: American Physiological Society
    Publication Date: 2016
    detail.hit.zdb_id: 1477297-8
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  • 6
    Online Resource
    Online Resource
    SerpinB1 deficiency is not associated with increased susceptibility to pulmonary emphysema in mice
    American Physiological Society ; 2013
    In:  American Journal of Physiology-Lung Cellular and Molecular Physiology Vol. 305, No. 12 ( 2013-12-15), p. L981-L989
    Details
    In: American Journal of Physiology-Lung Cellular and Molecular Physiology, American Physiological Society, Vol. 305, No. 12 ( 2013-12-15), p. L981-L989
    Abstract: Chronic obstructive pulmonary disease (COPD) is characterized by emphysema and chronic bronchitis and is a leading cause of morbidity and mortality worldwide. Tobacco smoke and deficiency in α1-antitrypsin (AAT) are the most prominent environmental and genetic risk factors, respectively. Yet the pathogenesis of COPD is not completely elucidated. Disease progression appears to include a vicious circle driven by self-perpetuating lung inflammation, endothelial and epithelial cell death, and proteolytic degradation of extracellular matrix proteins. Like AAT, serpinB1 is a potent inhibitor of serine proteases including neutrophil elastase and cathepsin G. Because serpinB1 is expressed in myeloid and lung epithelial cells and is protective during lung infections, we investigated the role of serpinB1 in preventing age-related and cigarette smoke-induced emphysema in mice. Fifteen-month-old mice showed increased lung volume and decreased pulmonary function compared with young adult mice (3 mo old), but no differences were observed between serpinB1-deficient (KO) and wild-type (WT) mice. Chronic exposure to secondhand cigarette smoke resulted in structural emphysematous changes compared with respective control mice, but no difference in lung morphometry was observed between genotypes. Of note, the different pattern of stereological changes induced by age and cigarette smoke suggest distinct mechanisms leading to increased airway volume. Finally, expression of intracellular and extracellular protease inhibitors were differently regulated in lungs of WT and KO mice following smoke exposure; however, activity of proteases was not significantly altered. In conclusion, we showed that, although AAT and serpinB1 are similarly potent inhibitors of neutrophil proteases, serpinB1 deficiency is not associated with more severe emphysema.
    Type of Medium: Online Resource
    ISSN: 1040-0605 , 1522-1504
    URL: Article
    DOI: 10.1152/ajplung.00181.2013
    Language: English
    Publisher: American Physiological Society
    Publication Date: 2013
    detail.hit.zdb_id: 1477300-4
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  • 7
    Online Resource
    Online Resource
    How common is the lipid body-containing interstitial cell in the mammalian lung?
    American Physiological Society ; 2014
    In:  American Journal of Physiology-Lung Cellular and Molecular Physiology Vol. 307, No. 5 ( 2014-09-01), p. L386-L394
    Details
    In: American Journal of Physiology-Lung Cellular and Molecular Physiology, American Physiological Society, Vol. 307, No. 5 ( 2014-09-01), p. L386-L394
    Abstract: Pulmonary lipofibroblasts are thought to be involved in lung development, regeneration, vitamin A storage, and surfactant synthesis. Most of the evidence for these important functions relies on mouse or rat studies. Therefore, the present study was designed to investigate the presence of lipofibroblasts in a variety of early postnatal and adult mammalian species (including humans) to evaluate the ability to generalize functions of this cell type for other species. For this purpose, lung samples from 14 adult mammalian species as well as from postnatal mice, rats, and humans were investigated using light and electron microscopic stereology to obtain the volume fraction and the total volume of lipid bodies. In adult animals, lipid bodies were observed only, but not in all rodents. In all other species, no lipofibroblasts were observed. In rodents, lipid body volume scaled with body mass with an exponent b = 0.73 in the power law equation. Lipid bodies were not observed in postnatal human lungs but showed a characteristic postnatal increase in mice and rats and persisted at a lower level in the adult animals. Among 14 mammalian species, lipofibroblasts were only observed in rodents. The great increase in lipid body volume during early postnatal development of the mouse lung confirms the special role of lipofibroblasts during rodent lung development. It is evident that the cellular functions of pulmonary lipofibroblasts cannot be transferred easily from rodents to other species, in particular humans.
    Type of Medium: Online Resource
    ISSN: 1040-0605 , 1522-1504
    URL: Article
    DOI: 10.1152/ajplung.00131.2014
    Language: English
    Publisher: American Physiological Society
    Publication Date: 2014
    detail.hit.zdb_id: 1477300-4
    SSG: 12
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  • 8
    Online Resource
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    Cutting-edge microangio-CT: new dimensions in vascular imaging and kidney morphometry
    American Physiological Society ; 2018
    In:  American Journal of Physiology-Renal Physiology Vol. 314, No. 3 ( 2018-03-01), p. F493-F499
    Details
    In: American Journal of Physiology-Renal Physiology, American Physiological Society, Vol. 314, No. 3 ( 2018-03-01), p. F493-F499
    Abstract: In the last decades, the contrast-enhanced micro-computed tomography (micro-CT) imaging of a whole animal kidney became increasingly important. The visualization was mainly limited to middle-sized vessels. Since modern desktop micro-CT scanners provide the necessary detail resolution, we developed an approach for rapid visualization and consistent assessment of kidney vasculature and glomeruli number. This method is based on μAngiofil, a new polymerizing contrast agent with homogenous X-ray absorption, which provides continuous filling of the complete vasculature and enables correlative imaging approaches. For rapid and reliable kidney morphometry, the microangio-CT (µaCT) data sets from glial cell line-derived neurotrophic factor (GDNF) +/− mice and their wild-type littermates were used. The results were obtained much faster compared with the current gold standard, histology-based stereology, and without processing artifacts. The histology-based morphometry was done afterward on the same kidneys. Both approaches revealed that the GDNF +/− male mice had about 40% fewer glomeruli. Furthermore, our approach allows for the definition of sites of interest for further histological investigation, i.e., correlative morphology. The polymerized μAngiofil stays in perfused vessels and is autofluorescent, which is what greatly facilitates the matching of histological sections with µaCT data. The presented approach is a time-efficient, reliable, qualitative, and quantitative methodology. Besides glomerular morphometry, the µaCT data can be used for qualitative and quantitative analysis of the kidney vasculature and correlative morphology.
    Type of Medium: Online Resource
    ISSN: 1931-857X , 1522-1466
    URL: Article
    DOI: 10.1152/ajprenal.00099.2017
    Language: English
    Publisher: American Physiological Society
    Publication Date: 2018
    detail.hit.zdb_id: 1477287-5
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  • 9
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    Diagnosis of primary ciliary dyskinesia: discrepancy according to different algorithms
    European Respiratory Society (ERS) ; 2021
    In:  ERJ Open Research Vol. 7, No. 4 ( 2021-10), p. 00353-2021-
    Details
    In: ERJ Open Research, European Respiratory Society (ERS), Vol. 7, No. 4 ( 2021-10), p. 00353-2021-
    Abstract: Diagnosis of primary ciliary dyskinesia (PCD) is challenging since there is no gold standard test. The European Respiratory (ERS) and American Thoracic (ATS) Societies developed evidence-based diagnostic guidelines with considerable differences. Objective We aimed to compare the algorithms published by the ERS and the ATS with each other and with our own PCD-UNIBE algorithm in a clinical setting. Our algorithm is similar to the ERS algorithm with additional immunofluorescence staining. Agreement (Cohen's κ) and concordance between the three algorithms were assessed in patients with suspicion of PCD referred to our diagnostic centre. Results In 46 out of 54 patients (85%) the final diagnosis was concordant between all three algorithms (30 PCD negative, 16 PCD positive). In eight patients (15%) PCD diagnosis differed between the algorithms. Five patients (9%) were diagnosed as PCD only by the ATS, one (2%) only by the ERS and PCD-UNIBE, one (2%) only by the ATS and PCD-UNIBE, and one (2%) only by the PCD-UNIBE algorithm. Agreement was substantial between the ERS and the ATS (κ=0.72, 95% CI 0.53–0.92) and the ATS and the PCD-UNIBE (κ=0.73, 95% CI 0.53–0.92) and almost perfect between the ERS and the PCD-UNIBE algorithms (κ=0.92, 95% CI 0.80–1.00). Conclusion The different diagnostic algorithms lead to a contradictory diagnosis in a considerable proportion of patients. Thus, an updated, internationally harmonised and standardised PCD diagnostic algorithm is needed to improve diagnostics for these discordant cases.
    Type of Medium: Online Resource
    ISSN: 2312-0541
    URL: Article
    DOI: 10.1183/23120541.00353-2021
    Language: English
    Publisher: European Respiratory Society (ERS)
    Publication Date: 2021
    detail.hit.zdb_id: 2827830-6
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  • 10
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    Online Resource
    Characterization of pediatric cystic fibrosis airway epithelial cell cultures at the air-liquid interface obtained by non-invasive nasal cytology brush sampling
    Springer Science and Business Media LLC ; 2017
    In:  Respiratory Research Vol. 18, No. 1 ( 2017-12)
    Details
    In: Respiratory Research, Springer Science and Business Media LLC, Vol. 18, No. 1 ( 2017-12)
    Type of Medium: Online Resource
    ISSN: 1465-993X
    URL: Article
    DOI: 10.1186/s12931-017-0706-7
    Language: English
    Publisher: Springer Science and Business Media LLC
    Publication Date: 2017
    detail.hit.zdb_id: 2041675-1
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