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Assessing the coagulation system in myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS)
Nunes, Jean Massimo. 2024. Assessing the coagulation system in myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS). Unpublished doctoral dissertation. Stellenbosch : Stellenbosch University [online]. Available: https://scholar.sun.ac.za/handle/10019.1/131870 Thesis (PhD)--Stellenbosch Univ...
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Stellenbosch : Stellenbosch University
2025
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| access_status_str | Open Access |
| author | Nunes, Jean Massimo |
| author2 | Pretorius, Resia |
| author_browse | Nunes, Jean Massimo Pretorius, Resia |
| author_facet | Pretorius, Resia Nunes, Jean Massimo |
| author_sort | Nunes, Jean Massimo |
| collection | Thesis |
| dc_rights_str_mv | Stellenbosch University |
| description | Nunes, Jean Massimo. 2024. Assessing the coagulation system in myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS). Unpublished doctoral dissertation. Stellenbosch : Stellenbosch University [online]. Available: https://scholar.sun.ac.za/handle/10019.1/131870
Thesis (PhD)--Stellenbosch University, 2024. |
| format | Thesis |
| id | oai:scholar.sun.ac.za:10019.1/131870 |
| institution | Stellenbosch University (South Africa) |
| last_indexed | 2026-06-10T12:44:24.894Z |
| license_str | Other — see source repository |
| provenance_str_mv | Harvested via OAI-PMH from SUNScholar — Stellenbosch University Repository |
| publishDate | 2025 |
| publishDateRange | 2025 |
| publishDateSort | 2025 |
| publisher | Stellenbosch : Stellenbosch University |
| publisherStr | Stellenbosch : Stellenbosch University |
| record_format | dspace |
| source_str | SUNScholar — Stellenbosch University Repository |
| spelling | oai:scholar.sun.ac.za:10019.1/131870 Assessing the coagulation system in myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) Nunes, Jean Massimo Pretorius, Resia Stellenbosch University. Faculty of Science. Dept. of Physiological Science. Myalgic encephalomyelitis Chronic fatigue syndrome Post COVID-19 condition (Disease) -- Alternative treatment Blood platelets -- Activation Blood -- Coagulation Thromboelastography Protein S Proteomics -- Therapeutic use UCTD Nunes, Jean Massimo. 2024. Assessing the coagulation system in myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS). Unpublished doctoral dissertation. Stellenbosch : Stellenbosch University [online]. Available: https://scholar.sun.ac.za/handle/10019.1/131870 Thesis (PhD)--Stellenbosch University, 2024. ENGLISH ABSTRACT: Introduction: Myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) is a debilitating and chronic post-viral disease that is characterized by unresolved fatigue, post-exertional symptom exacerbation (PESE), cognitive dysfunction, orthostatic intolerance, and gastrointestinal disturbances, among other symptoms. ME/CFS shares significant overlap with Long COVID (LC), the post-viral disease associated with SARS-CoV-2 infection, in both disease presentation and etiology. A prominent feature of LC pathology is a dysregulated coagulation system, characterized by anomalous clot formation, hyperactivated platelets, and microclots. In ME/CFS, the coagulation system is understudied, and hence represents a gap in knowledge. Therefore, this study aims to assess whether the clotting pathology present in LC is mirrored in ME/CFS. Methods: To assess the coagulation system in ME/CFS, 29 ME/CFS (22 females, 7 males, mean age of 45.7 ± 14.9) and 30 age- and gender-matched control (21 females, 9 males, mean age of 49.1 ± 11.3) blood samples were analyzed. Viscoelastic analysis of blood samples was conducted using thromboelastography (TEG®). Platelet activity was assessed via fluorescent microscopy and the use of two fluorescent markers specific for platelet activation markers, glycoprotein IIb/IIIa and P-selectin. Thioflavin T (ThT) was used to visualize microclots using fluorescent microscopy. Representative micrographs and ImageJ processing were used to infer, crudely, concentration values of microclots. To validate this assessment, microclot concentrations were measured by a recently established cell-free flow cytometry technique. To identify differentially expressed proteins, 15 randomly selected ME/CFS and 10 control PPP samples were subject to liquid chromatography tandem mass spectrometry (LC-MS/MS) analysis. Results: Findings from the TEG® assessment indicate that over half of the ME/CFS WB samples fell out of the standard clinical range, representative of a hypercoagulable profile. This inference is further supported by the analysis of PPP, where the greatest differences were recorded in α-angle (p=0.0006) and maximum rate to thrombus generation (p=0.0001). Roughly half of the ME/CFS cohort demonstrated platelet hyperactivity as determined by spreading, whereas only a quarter of the cohort was positive for significant platelet clumping. Using fluorescent microscopy and ImageJ software, it was inferred that the ME/CFS group contains more than 10x the levels of microclots (0.70 [11.21]) than the controls (0.06 [2.23]) (p<0.0001). Appropriate quantitative analysis with cell-free flow cytometry determined that the ME/CFS group exhibits a microclot burden 5x (27808 [107203]) that of the control group (4898 [20709]) (p<0.0001). Furthermore, ME/CFS PPP sample contain a greater prevalence of large microclots (≤100-400μm2). The proteomics analysis identified 45 differentially expressed proteins. Importantly, proteins related to clotting processes – thrombospondin-1, platelet factor 4, and protein S – were implicated. Complement machinery was also downregulated, whereas lactotransferrin and protein S100-A9 were upregulated. Conclusion: Overall, this study demonstrates that dysregulated clotting processes are an aspect of ME/CFS pathology, and that these abnormalities in coagulation are similar to that observed in LC. These findings provide further overlap between ME/CFS and LC, and has the potential to guide future research and prompt investigations into haematological pathology in ME/CFS. Importantly, this study highlights potential systems and proteins that require further research with regards to their contribution to the pathogenesis of ME/CFS, symptom manifestation, and biomarker potential, and also gives insight into the cardiovascular risk experienced by ME/CFS individuals. AFRIKAANSE OPSOMMING: Inleiding: Myalgiese enkefalomiëlitis/chroniese moegheidsindroom (ME/CFS) is 'n verswakkende en chroniese post-virale siekte wat gekenmerk word deur aanhoudende moegheid, simptoomverergering na inspanning (PESE), kognitiewe disfunksie, ortostatiese intoleransie, en maag-dermversteurings, onder andere simptome. ME/CFS het 'n beduidende oorvleueling met Lang COVID (LC), die post-virale siekte wat geassosieer word met SARS-CoV-2 infeksie, in beide siekte-aanbieding en etiologie. 'n Prominente kenmerk van LC-patologie is 'n gedisreguleerde stollingstelsel, gekenmerk deur abnormale klontvorming, hiperaktiewe bloedplaatjies, en mikroklonte. In ME/CFS is die stollingstelsel onderbestudeer, en verteenwoordig dus 'n gaping in kennis. Daarom is hierdie studie daarop gemik om te bepaal of die stollingspatologie wat in LC voorkom, ook in ME/CFS weerspieël word. Metodes: Om die stollingstelsel in ME/CFS te beoordeel, is 29 ME/CFS (22 vroue, 7 mans, gemiddelde ouderdom van 45.7 ± 14.9) en 30 ouderdom- en geslagsgelykstaande beheer (21 vroue, 9 mans, gemiddelde ouderdom van 49.1 ± 11.3) bloedmonsters ontleed. Viskoelastiese analise van bloedmonsters is uitgevoer met behulp van tromboelastografie (TEG®). Bloedplaatjie- aktiwiteit is beoordeel via fluoresserende mikroskopie en die gebruik van twee fluoresserende merkers spesifiek vir bloedplaatjie-aktiwasiemerker, glikoproteïen IIb/IIIa en P-selektien. Thioflavien T (ThT) is gebruik om mikroklonte met behulp van fluoresserende mikroskopie te visualiseer. Verteenwoordigende mikrofoto's en ImageJ-verwerking is gebruik om, grofweg, konsentrasiewaardes van mikroklonte af te lei. Om hierdie assessering te valideer, is mikroklontkonsentrasies gemeet deur 'n onlangs gevestigde selvrye vloeisitometrie tegniek, genaamd vloei-klotometrie. Om verskillend uitgedrukte proteïene te identifiseer, is 15 lukraak gekose ME/CFS en 10 beheer PPP monsters onderwerp aan vloeistofchromatografie tandem massaspektrometrie (LC-MS/MS) analise. Resultate: Bevindinge van die TEG®-assessering dui aan dat meer as die helfte van die ME/CFS WB-monsters buite die standaard kliniese reeks geval het, wat 'n hiperkoaguleerbare profiel verteenwoordig. Hierdie afleiding word verder ondersteun deur die analise van PPP, waar die grootste verskille opgeteken is in α-hoek (p=0.0006) en maksimum tempo tot trombusgenerasie (p=0.0001). Ongeveer die helfte van die ME/CFS-kohort het bloedplaatjiehiperaktiwiteit getoon soos bepaal deur verspreiding, terwyl slegs 'n kwart van die kohort positief was vir beduidende bloedplaatjiesamestelling. Met behulp van fluoresserende mikroskopie en ImageJ-sagteware is afgelei dat die ME/CFS-groep meer as 10x die vlakke van mikroklonte (0.70 [11.21]) bevat as die kontroles (0.06 [2.23]) (p<0.0001). Gepaste kwantitatiewe analise met vloei-klotometrie het bepaal dat die ME/CFS-groep 'n mikroklontlading van 5x (27808 [107203]) het vergeleke met die kontrolegroep (4898 [20709]) (p<0.0001). Verder bevat ME/CFS PPP-monsters 'n groter voorkoms van groot mikroklonte (≤100-400µm2). Die proteomika-analise het 45 verskillend uitgedrukte proteïene geïdentifiseer. Belangrik, proteïene wat verband hou met stollingsprosesse – trombospondien-1, bloedplaatjiefaktor 4, en proteïen S – was betrokke. Komplementmasjinerie was ook afgereguleer, terwyl laktoferrien en proteïen S100-A9 opgereguleer was. Gevolgtrekking: Algeheel toon hierdie studie dat gedisreguleerde stollingsprosesse 'n aspek van ME/CFS-patologie is en dat hierdie afwykings in stolling soortgelyk is aan dié wat in LC waargeneem word, hoewel minder ernstig. Hierdie bevindinge toon verdere oorvleueling tussen ME/CFS en LC, en het die potensiaal om toekomstige navorsing te rig en ondersoeke na hematologiese patologie in ME/CFS aan te moedig. Belangrik, hierdie studie beklemtoon potensiële stelsels en proteïene wat verdere navorsing vereis met betrekking tot hul bydrae tot die patogenese van ME/CFS, simptoommanifestasie, en biomerkerspotensiaal, en gee ook insig in die kardiovaskulêre risiko wat ME/CFS-individue ervaar. Doctoral 2025-04-04T09:13:22Z 2025-04-04T09:13:22Z 2024-12 Thesis https://scholar.sun.ac.za/handle/10019.1/131870 Stellenbosch University xix, 184 pages : illustrations application/pdf Stellenbosch : Stellenbosch University |
| spellingShingle | Myalgic encephalomyelitis Chronic fatigue syndrome Post COVID-19 condition (Disease) -- Alternative treatment Blood platelets -- Activation Blood -- Coagulation Thromboelastography Protein S Proteomics -- Therapeutic use UCTD Nunes, Jean Massimo Assessing the coagulation system in myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) |
| title | Assessing the coagulation system in myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) |
| title_full | Assessing the coagulation system in myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) |
| title_fullStr | Assessing the coagulation system in myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) |
| title_full_unstemmed | Assessing the coagulation system in myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) |
| title_short | Assessing the coagulation system in myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) |
| title_sort | assessing the coagulation system in myalgic encephalomyelitis chronic fatigue syndrome me cfs |
| topic | Myalgic encephalomyelitis Chronic fatigue syndrome Post COVID-19 condition (Disease) -- Alternative treatment Blood platelets -- Activation Blood -- Coagulation Thromboelastography Protein S Proteomics -- Therapeutic use UCTD |
| url | https://scholar.sun.ac.za/handle/10019.1/131870 |
| work_keys_str_mv | AT nunesjeanmassimo assessingthecoagulationsysteminmyalgicencephalomyelitischronicfatiguesyndromemecfs |