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  1. Article ; Online: Flow Cytometry of Male Reproductive Potential.

    Asosingh, Kewal

    Cytometry. Part A : the journal of the International Society for Analytical Cytology

    2020  Volume 97, Issue 12, Page(s) 1209–1210

    MeSH term(s) Flow Cytometry ; Humans ; Male ; Reproduction ; Spermatozoa
    Language English
    Publishing date 2020-07-13
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural
    ZDB-ID 2099868-5
    ISSN 1552-4930 ; 0196-4763 ; 1552-4922
    ISSN (online) 1552-4930
    ISSN 0196-4763 ; 1552-4922
    DOI 10.1002/cyto.a.24177
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    Zs.A 1688: Show issues Location:
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  2. Article ; Online: Hematopoietic stem cells and extramedullary hematopoiesis in the lungs.

    Reichard, Andrew / Wanner, Nicholas / Farha, Samar / Asosingh, Kewal

    Cytometry. Part A : the journal of the International Society for Analytical Cytology

    2023  Volume 103, Issue 12, Page(s) 967–977

    Abstract: Hematopoietic stem cells are key players in hematopoiesis as the body maintains a physiologic steady state, and the signaling pathways and control mechanisms of these dynamic cells are implicated in processes from inflammation to cancer. Although the ... ...

    Abstract Hematopoietic stem cells are key players in hematopoiesis as the body maintains a physiologic steady state, and the signaling pathways and control mechanisms of these dynamic cells are implicated in processes from inflammation to cancer. Although the bone marrow is commonly regarded as the site of hematopoiesis and hematopoietic stem cell residence, these cells also circulate in the blood and reside in extramedullary tissues, including the lungs. Flow cytometry is an invaluable tool in evaluating hematopoietic stem cells, revealing their phenotypes and relative abundances in both healthy and diseased states. This review outlines current protocols and cell markers used in flow cytometric analysis of hematopoietic stem and progenitor cell populations. Specific niches within the bone marrow are discussed, as are metabolic processes that contribute to stem cell self-renewal and differentiation, as well as the role of hematopoietic stem cells outside of the bone marrow at physiologic steady state. Finally, pulmonary extramedullary hematopoiesis and its associated disease states are outlined. Hematopoiesis in the lungs is a new and emerging concept, and discovering ways in which the study of lung-resident hematopoietic stem cells can be translated from murine models to patients will impact clinical treatment.
    MeSH term(s) Humans ; Animals ; Mice ; Hematopoiesis, Extramedullary ; Hematopoietic Stem Cells/metabolism ; Hematopoiesis ; Bone Marrow/metabolism ; Lung
    Language English
    Publishing date 2023-10-09
    Publishing country United States
    Document type Journal Article ; Review ; Research Support, N.I.H., Extramural
    ZDB-ID 2099868-5
    ISSN 1552-4930 ; 0196-4763 ; 1552-4922
    ISSN (online) 1552-4930
    ISSN 0196-4763 ; 1552-4922
    DOI 10.1002/cyto.a.24804
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  3. Article ; Online: Pathological Roles for Endothelial Colony-Forming Cells in Neonatal and Adult Lung Disease.

    Robertson, Jason O / Erzurum, Serpil C / Asosingh, Kewal

    American journal of respiratory cell and molecular biology

    2022  Volume 68, Issue 1, Page(s) 13–22

    Abstract: Endothelial colony-forming cells (ECFCs) are vascular resident and circulating endothelial cell subtypes with potent angiogenic capacity, a hierarchy of single-cell clonogenic potentials, and the ability to participate ... ...

    Abstract Endothelial colony-forming cells (ECFCs) are vascular resident and circulating endothelial cell subtypes with potent angiogenic capacity, a hierarchy of single-cell clonogenic potentials, and the ability to participate in
    MeSH term(s) Infant, Newborn ; Adult ; Humans ; Cells, Cultured ; Lung/pathology ; Endothelial Progenitor Cells ; Bronchopulmonary Dysplasia/pathology ; Infant, Premature ; Neovascularization, Physiologic ; Fetal Blood
    Language English
    Publishing date 2022-10-10
    Publishing country United States
    Document type Review ; Journal Article ; Research Support, N.I.H., Extramural
    ZDB-ID 1025960-0
    ISSN 1535-4989 ; 1044-1549
    ISSN (online) 1535-4989
    ISSN 1044-1549
    DOI 10.1165/rcmb.2022-0318PS
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  4. Article ; Online: Single-cell transcriptomic profiling of microvascular endothelial cell heterogeneity in congenital diaphragmatic hernia.

    Robertson, Jason O / Bazeley, Peter / Erzurum, Serpil C / Asosingh, Kewal

    Scientific reports

    2023  Volume 13, Issue 1, Page(s) 9851

    Abstract: Congenital diaphragmatic hernia (CDH) is a neonatal anomaly that includes pulmonary hypoplasia and hypertension. We hypothesized that microvascular endothelial cell (EC) heterogeneity is different in CDH lungs and related to lung underdevelopment and ... ...

    Abstract Congenital diaphragmatic hernia (CDH) is a neonatal anomaly that includes pulmonary hypoplasia and hypertension. We hypothesized that microvascular endothelial cell (EC) heterogeneity is different in CDH lungs and related to lung underdevelopment and remodeling. To test this, we evaluated rat fetuses at E21.5 in a nitrofen model of CDH to compare lung transcriptomes among healthy controls (2HC), nitrofen-exposed controls (NC) and nitrofen-exposed subjects with CDH. Single-cell RNA sequencing with unbiased clustering revealed 3 distinct microvascular EC clusters: a general population (mvEC), a proliferative population and a population high in hemoglobin. Only the CDH mvEC cluster had a distinct inflammatory transcriptomic signature as compared to the 2HC and NC endothelial cells, e.g. greater activation and adhesion of inflammatory cells and production of reactive oxygen species. Furthermore, CDH mvECs had downregulated Ca4, Apln and Ednrb gene expression. Those genes are markers for ECs important to lung development, gas exchange and alveolar repair (mvCa4+). mvCa4+ ECs were reduced in CDH (2HC [22.6%], NC [13.1%] and CDH [5.3%], p < 0.0001). Overall, these findings identify transcriptionally distinct microvascular endothelial cell clusters in CDH, including the distinctly inflammatory mvEC cluster and the depleted group of mvCa4+ ECs, which together may contribute to pathogenesis.
    MeSH term(s) Humans ; Rats ; Animals ; Hernias, Diaphragmatic, Congenital/genetics ; Hernias, Diaphragmatic, Congenital/pathology ; Endothelial Cells ; Transcriptome ; Rats, Sprague-Dawley ; Lung/pathology ; Disease Models, Animal
    Chemical Substances nitrofen (N71UYG034A)
    Language English
    Publishing date 2023-06-17
    Publishing country England
    Document type Journal Article
    ZDB-ID 2615211-3
    ISSN 2045-2322 ; 2045-2322
    ISSN (online) 2045-2322
    ISSN 2045-2322
    DOI 10.1038/s41598-023-37050-y
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  5. Article ; Online: Assessing the clinico-immunological profile of patients with obesity and chronic rhinosinusitis.

    Chaaban, Mohamad R / Asosingh, Kewal / Comhair, Suzy / Hoying, David

    International forum of allergy & rhinology

    2023  

    Abstract: Background: No studies have investigated the systemic and local sinonasal profile of obesity-related chronic rhinosinusitis (CRS), despite its observed association in recent retrospective studies. The objectives of our study were to assess the impact of ...

    Abstract Background: No studies have investigated the systemic and local sinonasal profile of obesity-related chronic rhinosinusitis (CRS), despite its observed association in recent retrospective studies. The objectives of our study were to assess the impact of obesity on the clinical and cytokine profile of patients with CRS and evaluate treatment response with functional endoscopic sinus surgery.
    Methods: This was a prospective observational study at the Cleveland Clinic that included patients with CRS (n = 54) between December 2021 and September 2022. Data collection included demographics, body mass index (BMI), comorbidities, baseline sinonasal outcome test scores, baseline radiologic scores (Lund-Mackay), postoperative sinonasal outcome test scores (at 3-4 months), and local and systemic alarmins/T-helper cytokines.
    Results: Out of the 54 CRS patients, there were 20 CRS patients without nasal polyps (37%) and 34 with nasal polyps (63%). Patients were categorized based on obesity (BMI ≥ 30 kg/m
    Conclusions: In conclusion, patients with obese CRS and nasal polyps displayed diminished levels of intranasal alarmins and reduced intranasal eotaxin-3. These results potentially imply the presence of a unique, obese type 2-low CRS phenotype that warrants further exploration.
    Language English
    Publishing date 2023-11-23
    Publishing country United States
    Document type Journal Article
    ZDB-ID 2625826-2
    ISSN 2042-6984 ; 2042-6976
    ISSN (online) 2042-6984
    ISSN 2042-6976
    DOI 10.1002/alr.23304
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  6. Article ; Online: Immunophenotyping of Circulating Endothelial Cells and Endothelial Microparticles.

    Wanner, Nicholas / Asosingh, Kewal

    Methods in molecular biology (Clifton, N.J.)

    2019  Volume 2032, Page(s) 203–211

    Abstract: Flow-cytometric detection of circulating endothelial cells and endothelial microparticles is an essential tool in studies of vascular diseases. Here we describe the principles and detailed methods for human blood sample processing, storage, labeling, and ...

    Abstract Flow-cytometric detection of circulating endothelial cells and endothelial microparticles is an essential tool in studies of vascular diseases. Here we describe the principles and detailed methods for human blood sample processing, storage, labeling, and gating of circulating endothelial elements.
    MeSH term(s) Blood Platelets/immunology ; Cell Count ; Cell-Derived Microparticles/immunology ; Endothelial Cells/immunology ; Flow Cytometry/methods ; Humans ; Immunophenotyping/methods ; Vascular Diseases/diagnosis ; Vascular Diseases/immunology
    Language English
    Publishing date 2019-08-08
    Publishing country United States
    Document type Journal Article
    ISSN 1940-6029
    ISSN (online) 1940-6029
    DOI 10.1007/978-1-4939-9650-6_12
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  7. Article: Using the Autofluorescence Finder on the Sony ID7000

    Wanner, Nicholas / Barnhart, Jerry / Apostolakis, Nicholas / Zlojutro, Violetta / Asosingh, Kewal

    Frontiers in bioengineering and biotechnology

    2022  Volume 10, Page(s) 827987

    Abstract: Autofluorescence (AF) is a feature of all cell types, though some have more than others. In tissues with complex heterogeneous cellularity, AF is frequently a source of high background, masking faint fluorescent signals and reducing the available dynamic ...

    Abstract Autofluorescence (AF) is a feature of all cell types, though some have more than others. In tissues with complex heterogeneous cellularity, AF is frequently a source of high background, masking faint fluorescent signals and reducing the available dynamic range of detectors for detecting fluorescence signals from markers of interest in a flow cytometry panel. Pulmonary flow cytometry presents unique challenges because lung cells are heterogeneous and contain varying amounts of high AF. The goal of this study was to demonstrate how a novel AF Finder tool on the Sony ID7000™ Spectral Cell Analyzer can be used to identify and screen multiple AF subsets in complex highly AF tissues like murine lungs. In lung single cell suspensions, the AF Finder tool identified four distinct AF spectra from six highly AF subsets. The subtraction of these distinct AF spectra resulted in a resolution increase by several log decades in several fluorescent channels. The major immune and lung tissue resident cells in a murine model of asthma were easily identified in a multi-color panel using AF subtraction. The findings demonstrate the practicality of the AF Finder tool, particularly when analyzing samples with multiple AF populations of varying intensities, in order to reduce fluorescence background and increase signal resolution in spectral flow cytometry.
    Language English
    Publishing date 2022-03-15
    Publishing country Switzerland
    Document type Journal Article
    ZDB-ID 2719493-0
    ISSN 2296-4185
    ISSN 2296-4185
    DOI 10.3389/fbioe.2022.827987
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  8. Article ; Online: Eosinophils: Ancient Cells with New Roles in Chronic Lung Inflammation.

    Asosingh, Kewal / Aronica, Mark A

    American journal of respiratory and critical care medicine

    2017  Volume 195, Issue 10, Page(s) 1281–1282

    Language English
    Publishing date 2017-05-15
    Publishing country United States
    Document type Journal Article
    ZDB-ID 1180953-x
    ISSN 1535-4970 ; 0003-0805 ; 1073-449X
    ISSN (online) 1535-4970
    ISSN 0003-0805 ; 1073-449X
    DOI 10.1164/rccm.201612-2435ED
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  9. Article ; Online: The role of mitochondria in angiogenesis.

    Reichard, Andrew / Asosingh, Kewal

    Molecular biology reports

    2018  Volume 46, Issue 1, Page(s) 1393–1400

    Abstract: Angiogenesis extends pre-existing blood vessels to improve oxygen and nutrient delivery to inflamed or otherwise hypoxic tissues. Mitochondria are integral in this process, controlling cellular metabolism to regulate the proliferation, migration, and ... ...

    Abstract Angiogenesis extends pre-existing blood vessels to improve oxygen and nutrient delivery to inflamed or otherwise hypoxic tissues. Mitochondria are integral in this process, controlling cellular metabolism to regulate the proliferation, migration, and survival of endothelial cells which comprise the inner lining of blood vessels. Mitochondrial Complex III senses hypoxic conditions and generates mitochondrial reactive oxygen species which stabilize hypoxia-inducible factor (HIF-1α) protein. HIF-1α induces the transcription of the vegfa gene, allowing the translation of vascular endothelial growth factor protein, which interacts with mature and precursor endothelial cells, mobilizing them to form new blood vessels. This cascade can be inhibited at specific points by means of gene knockdown, enzyme treatment, and introduction of naturally occurring small molecules, providing insight into the relationship between mitochondria and angiogenesis. This review focuses on current knowledge of the overall role of mitochondria in controlling angiogenesis and outlines known inhibitors that have been used to elucidate this pathway which may be useful in future research to control angiogenesis in vivo.
    MeSH term(s) Animals ; Cell Hypoxia ; Humans ; Mitochondria/metabolism ; Neovascularization, Physiologic ; Oxidation-Reduction ; Reactive Oxygen Species/metabolism ; Signal Transduction
    Chemical Substances Reactive Oxygen Species
    Language English
    Publishing date 2018-11-20
    Publishing country Netherlands
    Document type Journal Article ; Review
    ZDB-ID 186544-4
    ISSN 1573-4978 ; 0301-4851
    ISSN (online) 1573-4978
    ISSN 0301-4851
    DOI 10.1007/s11033-018-4488-x
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  10. Article ; Online: Best Practices for Preparing a Single Cell Suspension from Solid Tissues for Flow Cytometry.

    Reichard, Andrew / Asosingh, Kewal

    Cytometry. Part A : the journal of the International Society for Analytical Cytology

    2018  Volume 95, Issue 2, Page(s) 219–226

    Abstract: Preparing a single cell suspension is a critical step in any solid tissue flow cytometry experiment. Tissue dissection, enzymatic digestion, and mechanical dissociation are three significant steps leading to the degradation of the extracellular matrix ... ...

    Abstract Preparing a single cell suspension is a critical step in any solid tissue flow cytometry experiment. Tissue dissection, enzymatic digestion, and mechanical dissociation are three significant steps leading to the degradation of the extracellular matrix and the isolation of single cells, allowing the generation of high-quality flow cytometry data. Cells and the extracellular matrix contain various proteins and other structures which must be considered when designing a tissue digestion protocol to preserve the viability of cells and the presence of relevant antigens while digesting matrix components and cleaving cell-cell junctions. Evaluation of the single cell suspension is essential before proceeding with the labeling of the cells as high viability and absence of cell debris and aggregates are critical for flow cytometry. The information presented should be used as a general guide of steps to consider when preparing a single cell suspension from solid tissues for flow cytometry experiments. © 2018 International Society for Advancement of Cytometry.
    MeSH term(s) Animals ; Antigens/metabolism ; Cell Separation/methods ; Cell Survival/physiology ; Extracellular Matrix/metabolism ; Extracellular Matrix/physiology ; Flow Cytometry/methods ; Proteins/metabolism ; Single-Cell Analysis/methods
    Chemical Substances Antigens ; Proteins
    Language English
    Publishing date 2018-12-06
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural
    ZDB-ID 2099868-5
    ISSN 1552-4930 ; 0196-4763 ; 1552-4922
    ISSN (online) 1552-4930
    ISSN 0196-4763 ; 1552-4922
    DOI 10.1002/cyto.a.23690
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