Proteomic Evidence for Amyloidogenic Cross-Seeding in Fibrinaloid Microclots DOI Open Access
Douglas B. Kell, Etheresia Pretorius

International Journal of Molecular Sciences, Год журнала: 2024, Номер 25(19), С. 10809 - 10809

Опубликована: Окт. 8, 2024

In classical amyloidoses, amyloid fibres form through the nucleation and accretion of protein monomers, with protofibrils fibrils exhibiting a cross-β motif parallel or antiparallel β-sheets oriented perpendicular to fibre direction. These can intertwine mature fibres. Similar phenomena occur in blood from individuals circulating inflammatory molecules (and also some originating viruses bacteria). Such pathological clotting result an anomalous termed fibrinaloid microclots. Previous proteomic analyses these microclots have shown presence non-fibrin(ogen) proteins, suggesting more complex mechanism than simple entrapment. We thus provide evidence against such entrapment model, noting that clot pores are too large centrifugation would removed weakly bound proteins. Instead, we explore whether co-aggregation into may involve axial (multiple proteins within same fibril), lateral (single-protein contributing fibre), both types integration. Our analysis data different diseases shows no significant quantitative overlap normal plasma proteome correlation between abundance their Notably, abundant like α-2-macroglobulin, fibronectin, transthyretin absent microclots, while less as adiponectin, periostin, von Willebrand factor well represented. Using bioinformatic tools, including AmyloGram AnuPP, found entrapped exhibit high amyloidogenic tendencies, integration elements structures. This likely contributes microclots’ resistance proteolysis. findings underscore role cross-seeding microclot formation highlight need for further investigation structural properties implications thrombotic diseases. insights foundation developing novel diagnostic therapeutic strategies targeting disorders.

Язык: Английский

Automated microscopic measurement of fibrinaloid microclots and their degradation by nattokinase, the main natto protease DOI Creative Commons
Justine M. Grixti, Chrispian W. Theron, J. Enrique Salcedo-Sora

и другие.

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown

Опубликована: Апрель 7, 2024

Abstract Nattokinase, from the Japanese fermented food natto, is a protease with fibrinolytic activity that can thus degrade conventional blood clots. In some cases, however, including in Long COVID, fibrinogen polymerise into an anomalous amyloid form to create clots are resistant normal fibrinolysis and we refer as fibrinaloid microclots. These be detected fluorogenic stain thioflavin T. We describe automated microscopic technique for quantification of microclot formation, which also allows kinetics their formation aggregation recorded. here show recombinant nattokinase effective at degrading microclots vitro . This adds otherwise largely anecdotal evidence, review, might anticipated have value part therapeutic treatments individuals COVID related disorders involve

Язык: Английский

Процитировано

6
The Biology and Biochemistry of Kynurenic Acid, a Potential Nutraceutical with Multiple Biological Effects DOI Open Access
Luana de Fátima Alves, J. Bernadette Moore, Douglas B. Kell

и другие.

International Journal of Molecular Sciences, Год журнала: 2024, Номер 25(16), С. 9082 - 9082

Опубликована: Авг. 21, 2024

Kynurenic acid (KYNA) is an antioxidant degradation product of tryptophan that has been shown to have a variety cytoprotective, neuroprotective and neuronal signalling properties. However, mammalian transporters receptors display micromolar binding constants; these are consistent with its typically tissue concentrations but far above serum/plasma concentration (normally tens nanomolar), suggesting large gaps in our knowledge transport mechanisms action, the main influx characterized date equilibrative, not concentrative. In addition, it substrate known anion efflux pump (ABCC4), whose vivo activity largely unknown. Exogeneous addition L-tryptophan or L-kynurenine leads production KYNA also many other co-metabolites (including some such as 3-hydroxy-L-kynurenine quinolinic may be toxic). With exception chestnut honey, exists at relatively low levels natural foodstuffs. bioavailability reasonable, terminal element irreversible reaction most pathways, might added exogenously without disturbing upstream metabolism significantly. Many examples, which we review, show valuable bioactivity. Given above, review potential utility nutraceutical, finding significantly worthy further study development.

Язык: Английский

Процитировано

6
Treatment of Long COVID With Enoxaparin DOI

Ciara Wright,

Douglas B. Kell,

Resia Pretorius

и другие.

Cardiopulmonary Physical Therapy Journal, Год журнала: 2024, Номер 36(1), С. 70 - 73

Опубликована: Дек. 30, 2024

Background and Purpose: Long COVID is a complex multisystemic disease state, which represents huge economic health burden worldwide. Hypercoagulation the formation of fibrinaloid microclots have been proposed as an underlying pathology, may underpin wide variety presenting symptoms, via reduced blood flow cellular energy production. Postexertional malaise (PEM) common exercise therapy should not be recommended without addressing pathology. Case Presentation: Patient was female, 43, with for 21 months characterized primarily by extreme fatigue, intolerance, PEM, cognitive dysfunction, postural orthostatic tachycardia syndrome. She unable to work or leave house unless essential medical appointments. Before her initial severe acute respiratory syndrome coronavirus 2 infection, she had no prior conditions comorbidities, body mass index 20.8, engaged in regular vigorous exercise. Intervention: This case report describes patient treated enoxaparin, low molecular weight heparin, at 20 mg per day subcutaneously, assessment 8 weeks. Outcomes: The patient's function quality life improved markedly, reported subjectively, objective measurements Levels activity markedly adverse events. severity PEM decreased dramatically, better controlled. FUNCAP score, used measure functional capacity, increased from 3.3 (moderately affected) 5.07 (mildly after treatment. Discussion: It that this treatment further investigated elucidate role enoxaparin pathology COVID, including hypercoagulation microclots.

Язык: Английский

Процитировано

4
The Proteome Content of Blood Clots Observed Under Different Conditions: Successful Role in Predicting Clot Amyloid(ogenicity) DOI Creative Commons
Douglas B. Kell, Etheresia Pretorius

Molecules, Год журнала: 2025, Номер 30(3), С. 668 - 668

Опубликована: Фев. 3, 2025

A recent analysis compared the proteome of (i) blood clots seen in two diseases—sepsis and long COVID—when was known to have clotted into an amyloid microclot form (as judged by staining with fluorogenic stain thioflavin T) (ii) that those non-amyloid considered formed normally. Such fibrinaloid microclots are also relatively resistant fibrinolysis. The proteins contained differed markedly both from soluble typical plasma normal clots, between diseases studied (an acute syndrome sepsis ITU a chronic disease represented Long COVID). Many were low concentration effectively accumulated fibres, whereas many other abundant excluded. found associated tended be themselves amyloidogenic. We here ask inverse question. This is: can clot tell us whether particular observed uniquely (or highly over-represented) relative thus fact nature? answer is affirmative variety major coagulopathies, viz., venous thromboembolism, pulmonary embolism, deep vein thrombosis, various cardiac issues, ischaemic stroke. Galectin-3-binding protein thrombospondin-1 seem especially widely amyloid-type latter has indeed been shown incorporated growing fibrin fibres. These may consequently provide useful biomarkers mechanistic basis.

Язык: Английский

Процитировано

0
Oral SARS‐CoV‐2 Infection and Risk for Long Covid DOI Creative Commons

Joel Schwartz,

Kristelle J. Capistrano,

Heba Hussein

и другие.

Reviews in Medical Virology, Год журнала: 2025, Номер 35(2)

Опубликована: Март 1, 2025

ABSTRACT SARS‐CoV‐2 is an oral pathogen that infects and replicates in mucosal salivary epithelial cells, contributing to post‐acute sequelae COVID‐19 (PASC) other non‐oral pathologies. While pre‐existing inflammatory diseases provides a conducive environment for the virus, acute infection persistence of can also results microbiome dysbiosis further worsens poor health. Indeed, PASC includes periodontal diseases, dysgeusia, xerostomia, pharyngitis, keratoses, pulpitis suggesting significant bacterial contributions tissue tropism. Dysbiotic microbiome‐induced inflammation promote viral entry via angiotensin‐converting enzyme receptor‐2 (ACE2), serine transmembrane TMPRSS2 possibly non‐canonical pathways. Additionally, metabolites derived from dysbiotic alter physiological biochemical pathways related metabolism lipids, carbohydrates, amino acids. This may pro‐inflammatory microenvironment, leading immune exhaustion, loss tolerance, susceptibility variety pathogens, causing later chronic inflammation. Microbial release mimics host metallopeptidases furin, ADAM17 (A disintegrin metalloproteinase 17), glycoprotein aid attachment T cell immunoglobulin‐like (TIMs), enhancing while simultaneously depressing resistance clearance. Membrane reorganization characterised by neuroproteins, such as neuropilins, functionally assists with extends pathogenesis cavity brain, gut, or tissues. Thus, health, disrupted microbiomes tropism, weaken antiviral resistance, heightens infection. dysfunction increases risk additional infections, exacerbating conditions like endodontic diseases. These persistent health issues contribute systemic inflammation, creating bidirectional effects between tissues, potentially Post‐Acute Sequelae (PASC).

Язык: Английский

Процитировано

0
Proteomic evidence for amyloidogenic cross-seeding in fibrinaloid microclots DOI
Douglas B. Kell, Etheresia Pretorius

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown

Опубликована: Июль 17, 2024

Abstract In classical amyloidoses, amyloid fibres form through the nucleation and accretion of protein monomers, with protofibrils fibrils exhibiting a cross-β motif parallel or antiparallel β-sheets oriented perpendicular to fibre direction. These can intertwine mature fibres. Similar phenomena occur in blood from individuals circulating inflammatory molecules (also those originating viruses bacteria). presence inflammagens, pathological clotting occur, that results an anomalous termed fibrinaloid microclots. Previous proteomic analyses these microclots have shown non-fibrin(ogen) proteins, suggesting more complex mechanism than simple entrapment. We provide evidence against entrapment model, noting clot pores are too large centrifugation would removed weakly bound proteins. Instead, we explore whether co-aggregation into may involve axial (multiple proteins within same fibril), lateral (single-protein contributing fibre), both types integration. Our analysis data different diseases shows no significant overlap normal plasma proteome correlation between abundance Notably, abundant like α-2-macroglobulin, fibronectin, transthyretin absent microclots, while less such as adiponectin, periostin, von Willebrand Factor well represented. Using bioinformatic tools including AmyloGram AnuPP, found entrapped exhibit high amyloidogenic tendencies, their integration elements structures. This likely contributes microclots’ resistance proteolysis. findings underscore role cross-seeding microclot formation highlight need for further investigation structural properties implications thrombotic diseases. insights foundation developing novel diagnostic therapeutic strategies targeting disorders.

Язык: Английский

Процитировано

2
Flow Clotometry: Measuring Amyloid Microclots in ME/CFS, Long COVID, and Healthy Samples with Imaging Flow Cytometry DOI Creative Commons
Etheresia Pretorius,

Massimo Nunes,

Jan P Pretorius

и другие.

Research Square (Research Square), Год журнала: 2024, Номер unknown

Опубликована: Июнь 24, 2024

Abstract Myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) has received more attention since the characterization of Long COVID (LC), a condition somewhat similar in symptom presentation and, to some extent, pathophysiological mechanisms. A prominent feature LC pathology is amyloid, fibrinolysis-resistant fibrin(ogen) fragments, termed microclots. Despite prior identification microclots ME/CFS, quantitative analysis remained challenging due reliance on representative micrographs and software processing for estimations. Addressing this gap, present study uses cell-free imaging flow cytometry approach, optimized Thioflavin T-stained microclots, precisely measure microclot concentration size distribution across LC, healthy cohorts. We refer our technique detecting as 'flow clotometry'. demonstrate significant prevalence ME/CFS with patients exhibiting highest (18- 3-fold greater than groups, respectively). This finding underscores common both conditions, emphasizing dysregulated coagulation system. Moreover, relating distribution, group exhibited significantly higher all area ranges when compared controls, but demonstrated difference only single range group. suggests partially overlapping profile relative despite overall latter. The paves way prospective clinical application that aims efficiently detect, treat

Язык: Английский

Процитировано

1
Proteomic Evidence for Amyloidogenic Cross-Seeding in Fibrinaloid Microclots DOI Open Access
Douglas B. Kell, Etheresia Pretorius

International Journal of Molecular Sciences, Год журнала: 2024, Номер 25(19), С. 10809 - 10809

Опубликована: Окт. 8, 2024

In classical amyloidoses, amyloid fibres form through the nucleation and accretion of protein monomers, with protofibrils fibrils exhibiting a cross-β motif parallel or antiparallel β-sheets oriented perpendicular to fibre direction. These can intertwine mature fibres. Similar phenomena occur in blood from individuals circulating inflammatory molecules (and also some originating viruses bacteria). Such pathological clotting result an anomalous termed fibrinaloid microclots. Previous proteomic analyses these microclots have shown presence non-fibrin(ogen) proteins, suggesting more complex mechanism than simple entrapment. We thus provide evidence against such entrapment model, noting that clot pores are too large centrifugation would removed weakly bound proteins. Instead, we explore whether co-aggregation into may involve axial (multiple proteins within same fibril), lateral (single-protein contributing fibre), both types integration. Our analysis data different diseases shows no significant quantitative overlap normal plasma proteome correlation between abundance their Notably, abundant like α-2-macroglobulin, fibronectin, transthyretin absent microclots, while less as adiponectin, periostin, von Willebrand factor well represented. Using bioinformatic tools, including AmyloGram AnuPP, found entrapped exhibit high amyloidogenic tendencies, integration elements structures. This likely contributes microclots’ resistance proteolysis. findings underscore role cross-seeding microclot formation highlight need for further investigation structural properties implications thrombotic diseases. insights foundation developing novel diagnostic therapeutic strategies targeting disorders.

Язык: Английский

Процитировано

1