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  1. Article ; Online: Accumulation of Lipid Droplets Induced by Listeria monocytogenes in Macrophages: Implications for Survival and Evasion of Innate Immunity.

    Pereira-Dutra, Filipe S / Souza, Ellen K / Souza, Tamyris S / Goltara-Gomes, Taynná C / Ferraro-Moreira, Felipe / Palhinha, Lohanna / Cunha-Fernandes, Tamires / Rajão, Matheus A / Silva, Adriana R / Bozza, Patrícia T

    Journal of leukocyte biology

    2024  

    Abstract: Listeriosis, caused by Listeria monocytogenes (L.m.), poses a significant public health concern as one of the most severe foodborne diseases. The pathogenesis of L.m. involves critical steps such as phagosome rupture and escape upon internalization. ... ...

    Abstract Listeriosis, caused by Listeria monocytogenes (L.m.), poses a significant public health concern as one of the most severe foodborne diseases. The pathogenesis of L.m. involves critical steps such as phagosome rupture and escape upon internalization. Throughout infection, L.m. influences various host processes, including lipid metabolism pathways, yet the role of lipid droplets (LDs) remains unclear. Here, we reported a rapid, time-dependent increase in LD formation in macrophages induced by L.m. LD biogenesis was found to be dependent on L.m. viability and virulence genes, particularly on the activity of the pore-forming protein listeriolysin O (LLO). The prevention of LD formation by inhibiting diacylglycerol O-acyltransferase 1 (DGAT1) and cytosolic phospholipase A2 (cPLA2) significantly reduced intracellular bacterial survival, impaired prostaglandin E2 (PGE2) synthesis, and decreased IL-10 production. Additionally, inhibiting LD formation led to increased levels of TNF-α and IFN-β. Collectively, our data suggest a role for LDs in promoting L.m. cell survival and evasion within macrophages.
    Language English
    Publishing date 2024-05-10
    Publishing country England
    Document type Journal Article
    ZDB-ID 605722-6
    ISSN 1938-3673 ; 0741-5400
    ISSN (online) 1938-3673
    ISSN 0741-5400
    DOI 10.1093/jleuko/qiae115
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  2. Article ; Online: Density-based lipoprotein depletion improves extracellular vesicle isolation and functional analysis.

    Merij, Laura Botelho / da Silva, Luana Rocha / Palhinha, Lohanna / Gomes, Milena Tavares / Dib, Paula Ribeiro Braga / Martins-Gonçalves, Remy / Toledo-Quiroga, Kemily / Raposo-Nunes, Marcus Antônio / Andrade, Fernanda Brandi / de Toledo Martins, Sharon / Nascimento, Ana Lúcia Rosa / Rocha, Vinicius Novaes / Alves, Lysangela Ronalte / Bozza, Patrícia T / de Oliveira Trugilho, Monique Ramos / Hottz, Eugenio D

    Journal of thrombosis and haemostasis : JTH

    2024  Volume 22, Issue 5, Page(s) 1372–1388

    Abstract: Background: Blood plasma is the main source of extracellular vesicles (EVs) in clinical studies aiming to identify biomarkers and to investigate pathophysiological processes, especially regarding EV roles in inflammation and thrombosis. However, EV ... ...

    Abstract Background: Blood plasma is the main source of extracellular vesicles (EVs) in clinical studies aiming to identify biomarkers and to investigate pathophysiological processes, especially regarding EV roles in inflammation and thrombosis. However, EV isolation from plasma has faced the fundamental issue of lipoprotein contamination, representing an important bias since lipoproteins are highly abundant and modulate cell signaling, metabolism, and thromboinflammation.
    Objectives: Here, we aimed to isolate plasma EVs after depleting lipoproteins, thereby improving sample purity and EV thromboinflammatory analysis.
    Methods: Density-based gradient ultracentrifugation (G-UC) was used for lipoprotein depletion before EV isolation from plasma through size-exclusion chromatography (SEC) or serial centrifugation (SC). Recovered EVs were analyzed by size, concentration, cellular source, ultrastructure, and bottom-up proteomics.
    Results: G-UC efficiently separated lipoproteins from the plasma, allowing subsequent EV isolation through SEC or SC. Combined analysis from EV proteomics, cholesterol quantification, and apoB-100 detection confirmed the significant reduction in lipoproteins from isolated EVs. Proteomic analysis identified similar gene ontology and cellular components in EVs, regardless of lipoprotein depletion, which was consistent with similar EV cellular sources, size, and ultrastructure by flow cytometry and transmission electron microscopy. Importantly, lipoprotein depletion increased the detection of less abundant proteins in EV proteome and enhanced thromboinflammatory responses of platelets and monocytes stimulated in vitro with EV isolates.
    Conclusion: Combination of G-UC+SEC significantly reduced EV lipoprotein contamination without interfering in EV cellular source, gene ontology, and ultrastructure, allowing the recovery of highly pure EVs with potential implications for functional assays and proteomic and lipidomic analyses.
    MeSH term(s) Humans ; Extracellular Vesicles/metabolism ; Proteomics/methods ; Lipoproteins/blood ; Chromatography, Gel ; Blood Platelets/metabolism ; Centrifugation, Density Gradient ; Inflammation/blood ; Proteome ; Monocytes/metabolism
    Chemical Substances Lipoproteins ; Proteome
    Language English
    Publishing date 2024-01-24
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2112661-6
    ISSN 1538-7836 ; 1538-7933
    ISSN (online) 1538-7836
    ISSN 1538-7933
    DOI 10.1016/j.jtha.2024.01.010
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  3. Article ; Online: Leptin in the regulation of the immunometabolism of adipose tissue-macrophages.

    Monteiro, Lauar / Pereira, Jéssica Aparecida da Silva / Palhinha, Lohanna / Moraes-Vieira, Pedro Manoel M

    Journal of leukocyte biology

    2019  Volume 106, Issue 3, Page(s) 703–716

    Abstract: Obesity is a pandemic disease affecting around 15% of the global population. Obesity is a major risk factor for other conditions, such as type 2 diabetes and cardiovascular diseases. The adipose tissue is the main secretor of leptin, an adipokine ... ...

    Abstract Obesity is a pandemic disease affecting around 15% of the global population. Obesity is a major risk factor for other conditions, such as type 2 diabetes and cardiovascular diseases. The adipose tissue is the main secretor of leptin, an adipokine responsible for the regulation of food intake and energy expenditure. Obese individuals become hyperleptinemic due to increased adipogenesis. Leptin acts through the leptin receptor and induces several immunometabolic changes in different cell types, including adipocytes and Mϕs. Adipose tissue resident Mϕs (ATMs) are the largest leukocyte population in the adipose tissue and these ATMs are in constant contact with the excessive leptin levels secreted in obese conditions. Leptin activates both the JAK2-STAT3 and the PI3K-AKT-mTOR pathways. The activation of these pathways leads to intracellular metabolic changes, with increased glucose uptake, upregulation of glycolytic enzymes, and disruption of mitochondrial function, as well as immunologic alterations, such as increased phagocytic activity and proinflammatory cytokines secretion. Here, we discuss the immunometabolic effects of leptin in Mϕs and how hyperleptinemia can contribute to the low-grade systemic inflammation in obesity.
    MeSH term(s) Adipose Tissue/cytology ; Animals ; Humans ; Immunity ; Leptin/metabolism ; Macrophages/immunology ; Macrophages/metabolism ; Receptors, Leptin/metabolism ; Signal Transduction
    Chemical Substances Leptin ; Receptors, Leptin
    Language English
    Publishing date 2019-05-14
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't ; Review
    ZDB-ID 605722-6
    ISSN 1938-3673 ; 0741-5400
    ISSN (online) 1938-3673
    ISSN 0741-5400
    DOI 10.1002/JLB.MR1218-478R
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  4. Article ; Online: Generalized Edema and Pseudothrombocytopenia After ChAdOx1 nCoV-19 COVID-19 Vaccination: A Case Report.

    Bokel, Joanna / Mendes-de-Almeida, Daniela P / Martins-Gonçalves, Remy / Palhinha, Lohanna / Vizzoni, Alexandre G / Correa, Danusa Ferreira / Brandão, Luciana Gomes Pedro / Bozza, Patrícia T / Grinsztejn, Beatriz

    Frontiers in public health

    2022  Volume 10, Page(s) 907652

    Abstract: Reports of side effects of vaccines against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are increasing worldwide. Capillary leak syndrome and vaccine-induced immune thrombotic thrombocytopenia are very rare but life-threatening adverse ... ...

    Abstract Reports of side effects of vaccines against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are increasing worldwide. Capillary leak syndrome and vaccine-induced immune thrombotic thrombocytopenia are very rare but life-threatening adverse events that should be identified early and treated. However, isolated thrombocytopenia can indicate pseudothrombocytopenia. In certain people, ethylenediaminetetraacetic acid (EDTA) induces an
    MeSH term(s) COVID-19/prevention & control ; COVID-19 Vaccines/adverse effects ; ChAdOx1 nCoV-19 ; Edema ; Humans ; SARS-CoV-2 ; Thrombocytopenia/chemically induced ; Thrombocytopenia/diagnosis ; Vaccination/adverse effects
    Chemical Substances COVID-19 Vaccines ; ChAdOx1 nCoV-19 (B5S3K2V0G8)
    Language English
    Publishing date 2022-05-27
    Publishing country Switzerland
    Document type Case Reports ; Research Support, Non-U.S. Gov't
    ZDB-ID 2711781-9
    ISSN 2296-2565 ; 2296-2565
    ISSN (online) 2296-2565
    ISSN 2296-2565
    DOI 10.3389/fpubh.2022.907652
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  5. Article ; Online: Persisting Platelet Activation and Hyperactivity in COVID-19 Survivors.

    Martins-Gonçalves, Remy / Campos, Mariana M / Palhinha, Lohanna / Azevedo-Quintanilha, Isaclaudia G / Abud Mendes, Mayara / Ramos Temerozo, Jairo / Toledo-Mendes, Júlia / Rosado-de-Castro, Paulo H / Bozza, Fernando A / Souza Rodrigues, Rosana / Hottz, Eugenio D / Bozza, Patricia T

    Circulation research

    2022  Volume 131, Issue 11, Page(s) 944–947

    MeSH term(s) Humans ; COVID-19 ; Platelet Activation ; SARS-CoV-2 ; Survivors ; Blood Platelets/physiology
    Language English
    Publishing date 2022-10-21
    Publishing country United States
    Document type Letter ; Research Support, Non-U.S. Gov't
    ZDB-ID 80100-8
    ISSN 1524-4571 ; 0009-7330 ; 0931-6876
    ISSN (online) 1524-4571
    ISSN 0009-7330 ; 0931-6876
    DOI 10.1161/CIRCRESAHA.122.321659
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  6. Article ; Online: Platelet activation and platelet-monocyte aggregate formation trigger tissue factor expression in patients with severe COVID-19.

    Hottz, Eugenio D / Azevedo-Quintanilha, Isaclaudia G / Palhinha, Lohanna / Teixeira, Lívia / Barreto, Ester A / Pão, Camila R R / Righy, Cassia / Franco, Sérgio / Souza, Thiago M L / Kurtz, Pedro / Bozza, Fernando A / Bozza, Patrícia T

    Blood

    2020  Volume 136, Issue 11, Page(s) 1330–1341

    Abstract: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is an emergent pathogen responsible for the coronavirus disease 2019 (COVID-19). Since its emergence, the novel coronavirus has rapidly achieved pandemic proportions causing remarkably ... ...

    Abstract Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is an emergent pathogen responsible for the coronavirus disease 2019 (COVID-19). Since its emergence, the novel coronavirus has rapidly achieved pandemic proportions causing remarkably increased morbidity and mortality around the world. A hypercoagulability state has been reported as a major pathologic event in COVID-19, and thromboembolic complications listed among life-threatening complications of the disease. Platelets are chief effector cells of hemostasis and pathological thrombosis. However, the participation of platelets in the pathogenesis of COVID-19 remains elusive. This report demonstrates that increased platelet activation and platelet-monocyte aggregate formation are observed in severe COVID-19 patients, but not in patients presenting mild COVID-19 syndrome. In addition, exposure to plasma from severe COVID-19 patients increased the activation of control platelets ex vivo. In our cohort of COVID-19 patients admitted to the intensive care unit, platelet-monocyte interaction was strongly associated with tissue factor (TF) expression by the monocytes. Platelet activation and monocyte TF expression were associated with markers of coagulation exacerbation as fibrinogen and D-dimers, and were increased in patients requiring invasive mechanical ventilation or patients who evolved with in-hospital mortality. Finally, platelets from severe COVID-19 patients were able to induce TF expression ex vivo in monocytes from healthy volunteers, a phenomenon that was inhibited by platelet P-selectin neutralization or integrin αIIb/β3 blocking with the aggregation inhibitor abciximab. Altogether, these data shed light on new pathological mechanisms involving platelet activation and platelet-dependent monocyte TF expression, which were associated with COVID-19 severity and mortality.
    MeSH term(s) Adult ; Betacoronavirus/immunology ; Biomarkers/metabolism ; Blood Coagulation Disorders/immunology ; Blood Coagulation Disorders/metabolism ; Blood Coagulation Disorders/pathology ; Blood Coagulation Disorders/virology ; Blood Platelets/metabolism ; Blood Platelets/pathology ; Blood Platelets/virology ; COVID-19 ; Case-Control Studies ; Coronavirus Infections/complications ; Coronavirus Infections/immunology ; Coronavirus Infections/metabolism ; Coronavirus Infections/virology ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Monocytes/metabolism ; Monocytes/pathology ; Monocytes/virology ; P-Selectin/metabolism ; Pandemics ; Platelet Activation ; Pneumonia, Viral/complications ; Pneumonia, Viral/immunology ; Pneumonia, Viral/metabolism ; Pneumonia, Viral/virology ; Prognosis ; Prospective Studies ; SARS-CoV-2 ; Survival Rate ; Thromboplastin/metabolism
    Chemical Substances Biomarkers ; P-Selectin ; SELP protein, human ; Thromboplastin (9035-58-9)
    Keywords covid19
    Language English
    Publishing date 2020-07-17
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 80069-7
    ISSN 1528-0020 ; 0006-4971
    ISSN (online) 1528-0020
    ISSN 0006-4971
    DOI 10.1182/blood.2020007252
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  7. Article: A Rare Potential Pathogenic Variant in the

    da Fonseca, Ana Carolina Proença / Abreu, Gabriella de Medeiros / Palhinha, Lohanna / Zembrzuski, Verônica Marques / Campos Junior, Mario / Carneiro, João Regis Ivar / Nogueira Neto, José Firmino / Magno, Fernanda Cristina C Mattos / Rosado, Eliane Lopes / Maya-Monteiro, Clarissa Menezes / de Cabello, Giselda Maria Kalil / Cabello, Pedro Hernán / Bozza, Patricia Torres

    Diabetes, metabolic syndrome and obesity : targets and therapy

    2021  Volume 14, Page(s) 11–22

    Abstract: Background: Brain-derived neurotrophic factor (BDNF) is a pro-survival factor in the brain that also regulates energy balance. : Material and methods: This study comprised 201 adults with severe obesity (BMI ≥ 35.0 kg/m: Results: As a result, ... ...

    Abstract Background: Brain-derived neurotrophic factor (BDNF) is a pro-survival factor in the brain that also regulates energy balance.
    Material and methods: This study comprised 201 adults with severe obesity (BMI ≥ 35.0 kg/m
    Results: As a result, three missense variants [p.(Thr2Ile), p.(Val66Met), and p.(Arg209Gln)] and four synonymous variants (p.Leu107=, p.Thr149=, p.Ala150=, and p.Ser213=) were identified. The p.(Arg209Gln) was predicted as pathogenic by all in silico algorithms used and was not observed in the control group. The individuals carrying the p.(Val66Met) mutated allele had higher waist circumference, HDL-cholesterol and MCP1 levels, and reduced risk of developing metabolic syndrome.
    Conclusion: We observed that the common
    Language English
    Publishing date 2021-01-06
    Publishing country New Zealand
    Document type Journal Article
    ZDB-ID 2494854-8
    ISSN 1178-7007
    ISSN 1178-7007
    DOI 10.2147/DMSO.S267202
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  8. Article ; Online: Intracerebral hemorrhage associated with vaccine-induced thrombotic thrombocytopenia following ChAdOx1 nCOVID-19 vaccine in a pregnant woman.

    Mendes-de-Almeida, Daniela P / Martins-Gonçalves, Remy / Morato-Santos, Renata / De Carvalho, Gustavo Adolpho C / Martins, Silas A / Palhinha, Lohanna / Sandim, Vanessa / Avvad-Portari, Elyzabeth / Bozza, Fernando A / Monteiro, Robson Q / Bozza, Patrícia T / Kurtz, Pedro

    Haematologica

    2021  Volume 106, Issue 11, Page(s) 3025–3028

    MeSH term(s) COVID-19 Vaccines/adverse effects ; Cerebral Hemorrhage/etiology ; Female ; Humans ; Pregnancy ; Pregnant Women ; Thrombocytopenia/chemically induced ; Vaccines/adverse effects
    Chemical Substances COVID-19 Vaccines ; Vaccines
    Language English
    Publishing date 2021-11-01
    Publishing country Italy
    Document type Journal Article
    ZDB-ID 2333-4
    ISSN 1592-8721 ; 0017-6567 ; 0390-6078
    ISSN (online) 1592-8721
    ISSN 0017-6567 ; 0390-6078
    DOI 10.3324/haematol.2021.279407
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  9. Article ; Online: SH2B1 variants as potential causes of non-syndromic monogenic obesity in a Brazilian cohort.

    da Fonseca, Ana Carolina Proença / Assis, Izadora Sthephanie Silva / Salum, Kaio Cezar Rodrigues / Palhinha, Lohanna / Abreu, Gabriella Medeiros / Zembrzuski, Verônica Marques / Campos Junior, Mario / Nogueira Neto, José Firmino / Mattos, Fernanda Cristina C / Cambraia, Amanda / Rosado, Eliane Lopes / Maya-Monteiro, Clarissa Menezes / Cabello, Pedro Hernán / Carneiro, João Regis Ivar / Bozza, Patrícia T

    Eating and weight disorders : EWD

    2022  

    Abstract: Purpose: SH2B1 gene encodes an important adaptor protein to receptor tyrosine kinases or cytokine receptors associated with Janus kinases. This gene has been associated with the structural and functional modulation of neurons and other cells, and ... ...

    Abstract Purpose: SH2B1 gene encodes an important adaptor protein to receptor tyrosine kinases or cytokine receptors associated with Janus kinases. This gene has been associated with the structural and functional modulation of neurons and other cells, and impacts on energy and glucose homeostasis. Several studies suggested that alterations in this gene are strong candidates for the development of obesity. However, only a few studies have screened SH2B1 point variants in individuals with obesity. Therefore, the aim of this study was to investigate the prevalence of SH2B1 variants in a Brazilian cohort of patients with severe obesity and candidates to bariatric surgery.
    Methods: The cohort comprised 122 individuals with severe obesity, who developed this phenotype during childhood. As controls, 100 normal-weight individuals were included. The coding region of SH2B1 gene was screened by Sanger sequencing.
    Results: A total of eight variants were identified in SH2B1, of which p.(Val345Met) and p.(Arg630Gln) variants were rare and predicted as potentially pathogenic by the in the silico algorithms used in this study. The p.(Val345Met) was not found in either the control group or in publicly available databases. This variant was identified in a female patient with severe obesity, metabolic syndrome and hyperglycemia. The p.(Arg630Gln) was also absent in our control group, but it was reported in gnomAD with an extremely low frequency. This variant was observed in a female patient with morbid obesity, metabolic syndrome, hypertension and severe binge-eating disorder.
    Conclusion: Our study reported for the first time two rare and potentially pathogenic variants in Brazilian patients with severe obesity. Further functional studies will be necessary to confirm and elucidate the impact of these variants on SH2B1 protein function and stability, and their impact on energetic metabolism.
    Level of evidence: Level V, cross-sectional descriptive study.
    Language English
    Publishing date 2022-11-27
    Publishing country Germany
    Document type Journal Article
    ZDB-ID 2038625-4
    ISSN 1590-1262 ; 1124-4909
    ISSN (online) 1590-1262
    ISSN 1124-4909
    DOI 10.1007/s40519-022-01506-3
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  10. Article ; Online: Platelet-monocyte interaction amplifies thromboinflammation through tissue factor signaling in COVID-19.

    Hottz, Eugenio D / Martins-Gonçalves, Remy / Palhinha, Lohanna / Azevedo-Quintanilha, Isaclaudia G / de Campos, Mariana M / Sacramento, Carolina Q / Temerozo, Jairo R / Soares, Vinicius Cardoso / Dias, Suelen S Gomes / Teixeira, Lívia / Castro, Ícaro / Righy, Cassia / Souza, Thiago Moreno L / Kurtz, Pedro / Andrade, Bruno B / Nakaya, Helder I / Monteiro, Robson Q / Bozza, Fernando A / Bozza, Patrícia T

    Blood advances

    2022  Volume 6, Issue 17, Page(s) 5085–5099

    Abstract: Accumulating evidence into the pathogenesis of COVID-19 highlights a hypercoagulability state with high risk of life-threatening thromboembolic complications. However, the mechanisms of hypercoagulability and their link to hyperinflammation remain poorly ...

    Abstract Accumulating evidence into the pathogenesis of COVID-19 highlights a hypercoagulability state with high risk of life-threatening thromboembolic complications. However, the mechanisms of hypercoagulability and their link to hyperinflammation remain poorly understood. Here, we investigate functions and mechanisms of platelet activation and platelet-monocyte interactions in inflammatory amplification during SARS-CoV-2 infection. We used a combination of immunophenotyping, single-cell analysis, functional assays, and pharmacological approaches to gain insights on mechanisms. Critically ill patients with COVID-19 exhibited increased platelet-monocyte aggregates formation. We identified a subset of inflammatory monocytes presenting high CD16 and low HLA-DR expression as the subset mainly interacting with platelets during severe COVID-19. Single-cell RNA-sequencing analysis indicated enhanced fibrinogen receptor Mac-1 in monocytes from patients with severe COVID-19. Monocytes from patients with severe COVID-19 displayed increased platelet binding and hyperresponsiveness to P-selectin and fibrinogen with respect to tumor necrosis factor-α and interleukin-1β secretion. Platelets were able to orchestrate monocyte responses driving tissue factor (TF) expression, inflammatory activation, and inflammatory cytokines secretion in SARS-CoV-2 infection. Platelet-monocyte interactions ex vivo and in SARS-CoV-2 infection model in vitro reciprocally activated monocytes and platelets, inducing the heightened secretion of a wide panel of inflammatory mediators. We identified platelet adhesion as a primary signaling mechanism inducing mediator secretion and TF expression, whereas TF signaling played major roles in amplifying inflammation by inducing proinflammatory cytokines, especially tumor necrosis factor-α and interleukin-1β. Our data identify platelet-induced TF expression and activity at the crossroad of coagulation and inflammation in severe COVID-19.
    MeSH term(s) Blood Platelets/metabolism ; COVID-19 ; Cytokines/metabolism ; Humans ; Inflammation/pathology ; Interleukin-1beta/metabolism ; Monocytes/metabolism ; SARS-CoV-2 ; Thromboinflammation ; Thrombophilia ; Thromboplastin/metabolism ; Thrombosis/metabolism ; Tumor Necrosis Factor-alpha/metabolism
    Chemical Substances Cytokines ; Interleukin-1beta ; Tumor Necrosis Factor-alpha ; Thromboplastin (9035-58-9)
    Language English
    Publishing date 2022-04-14
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2915908-8
    ISSN 2473-9537 ; 2473-9529
    ISSN (online) 2473-9537
    ISSN 2473-9529
    DOI 10.1182/bloodadvances.2021006680
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