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Article ; Online: Prevalence of readily detected amyloid blood clots in 'unclotted' Type 2 Diabetes Mellitus and COVID-19 plasma: a preliminary report.

Pretorius, Etheresia / Venter, Chantelle / Laubscher, Gert Jacobus / Lourens, Petrus Johannes / Steenkamp, Janami / Kell, Douglas B

Cardiovascular diabetology

2020 Volume 19, Issue 1, Page(s) 193

Abstract: Background: Type 2 Diabetes Mellitus (T2DM) is a well-known comorbidity to COVID-19 and coagulopathies are a common accompaniment to both T2DM and COVID-19. In addition, patients with COVID-19 are known to develop micro-clots within the lungs. The rapid ...

Abstract	Background: Type 2 Diabetes Mellitus (T2DM) is a well-known comorbidity to COVID-19 and coagulopathies are a common accompaniment to both T2DM and COVID-19. In addition, patients with COVID-19 are known to develop micro-clots within the lungs. The rapid detection of COVID-19 uses genotypic testing for the presence of SARS-Cov-2 virus in nasopharyngeal swabs, but it can have a poor sensitivity. A rapid, host-based physiological test that indicated clotting severity and the extent of clotting pathologies in the individual who was infected or not would be highly desirable. Methods: Platelet poor plasma (PPP) was collected and frozen. On the day of analysis, PPP samples were thawed and analysed. We show here that microclots can be detected in the native plasma of twenty COVID-19, as well as ten T2DM patients, without the addition of any clotting agent, and in particular that such clots are amyloid in nature as judged by a standard fluorogenic stain. Results were compared to ten healthy age-matched individuals. Results: In COVID-19 plasma these microclots are significantly increased when compared to the levels in T2DM. Conclusions: This fluorogenic test may provide a rapid and convenient test with 100% sensitivity (P < 0.0001) and is consistent with the recognition that the early detection and prevention of such clotting can have an important role in therapy.
MeSH term(s)	Amyloid/blood ; COVID-19/blood ; COVID-19/epidemiology ; Diabetes Mellitus, Type 2/blood ; Diabetes Mellitus, Type 2/epidemiology ; Female ; Humans ; Male ; Middle Aged ; Prevalence ; SARS-CoV-2 ; Thrombosis/blood ; Thrombosis/epidemiology
Chemical Substances	Amyloid
Keywords	covid19
Language	English
Publishing date	2020-11-17
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ISSN	1475-2840
ISSN (online)	1475-2840
DOI	10.1186/s12933-020-01165-7
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: SARS-CoV-2 spike protein S1 induces fibrin(ogen) resistant to fibrinolysis: implications for microclot formation in COVID-19.

Grobbelaar, Lize M / Venter, Chantelle / Vlok, Mare / Ngoepe, Malebogo / Laubscher, Gert Jacobus / Lourens, Petrus Johannes / Steenkamp, Janami / Kell, Douglas B / Pretorius, Etheresia

Bioscience reports

2021 Volume 41, Issue 8

Abstract: Severe acute respiratory syndrome coronavirus 2 (SARS-Cov-2)-induced infection, the cause of coronavirus disease 2019 (COVID-19), is characterized by unprecedented clinical pathologies. One of the most important pathologies, is hypercoagulation and ... ...

Abstract	Severe acute respiratory syndrome coronavirus 2 (SARS-Cov-2)-induced infection, the cause of coronavirus disease 2019 (COVID-19), is characterized by unprecedented clinical pathologies. One of the most important pathologies, is hypercoagulation and microclots in the lungs of patients. Here we study the effect of isolated SARS-CoV-2 spike protein S1 subunit as potential inflammagen sui generis. Using scanning electron and fluorescence microscopy as well as mass spectrometry, we investigate the potential of this inflammagen to interact with platelets and fibrin(ogen) directly to cause blood hypercoagulation. Using platelet-poor plasma (PPP), we show that spike protein may interfere with blood flow. Mass spectrometry also showed that when spike protein S1 is added to healthy PPP, it results in structural changes to β and γ fibrin(ogen), complement 3, and prothrombin. These proteins were substantially resistant to trypsinization, in the presence of spike protein S1. Here we suggest that, in part, the presence of spike protein in circulation may contribute to the hypercoagulation in COVID-19 positive patients and may cause substantial impairment of fibrinolysis. Such lytic impairment may result in the persistent large microclots we have noted here and previously in plasma samples of COVID-19 patients. This observation may have important clinical relevance in the treatment of hypercoagulability in COVID-19 patients.
MeSH term(s)	Adult ; Aged ; Amyloid/metabolism ; Blood Platelets/metabolism ; COVID-19/pathology ; Complement C3/metabolism ; Female ; Fibrin/metabolism ; Fibrinogen/metabolism ; Fibrinolysis/physiology ; Humans ; Lung/pathology ; Male ; Microfluidic Analytical Techniques ; Middle Aged ; Prothrombin/metabolism ; SARS-CoV-2/metabolism ; Spike Glycoprotein, Coronavirus/metabolism ; Thrombosis/pathology ; Thrombosis/virology ; Trypsin/metabolism
Chemical Substances	Amyloid ; Complement C3 ; Spike Glycoprotein, Coronavirus ; spike protein, SARS-CoV-2 ; Prothrombin (9001-26-7) ; Fibrin (9001-31-4) ; Fibrinogen (9001-32-5) ; Trypsin (EC 3.4.21.4)
Language	English
Publishing date	2021-07-30
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	764946-0
ISSN	1573-4935 ; 0144-8463
ISSN (online)	1573-4935
ISSN	0144-8463
DOI	10.1042/BSR20210611
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Article ; Online: Persistent clotting protein pathology in Long COVID/Post-Acute Sequelae of COVID-19 (PASC) is accompanied by increased levels of antiplasmin.

Pretorius, Etheresia / Vlok, Mare / Venter, Chantelle / Bezuidenhout, Johannes A / Laubscher, Gert Jacobus / Steenkamp, Janami / Kell, Douglas B

Cardiovascular diabetology

2021 Volume 20, Issue 1, Page(s) 172

Abstract: Background: Severe acute respiratory syndrome coronavirus 2 (SARS-Cov-2)-induced infection, the cause of coronavirus disease 2019 (COVID-19), is characterized by acute clinical pathologies, including various coagulopathies that may be accompanied by ... ...

Abstract	Background: Severe acute respiratory syndrome coronavirus 2 (SARS-Cov-2)-induced infection, the cause of coronavirus disease 2019 (COVID-19), is characterized by acute clinical pathologies, including various coagulopathies that may be accompanied by hypercoagulation and platelet hyperactivation. Recently, a new COVID-19 phenotype has been noted in patients after they have ostensibly recovered from acute COVID-19 symptoms. This new syndrome is commonly termed Long COVID/Post-Acute Sequelae of COVID-19 (PASC). Here we refer to it as Long COVID/PASC. Lingering symptoms persist for as much as 6 months (or longer) after acute infection, where COVID-19 survivors complain of recurring fatigue or muscle weakness, being out of breath, sleep difficulties, and anxiety or depression. Given that blood clots can block microcapillaries and thereby inhibit oxygen exchange, we here investigate if the lingering symptoms that individuals with Long COVID/PASC manifest might be due to the presence of persistent circulating plasma microclots that are resistant to fibrinolysis. Methods: We use techniques including proteomics and fluorescence microscopy to study plasma samples from healthy individuals, individuals with Type 2 Diabetes Mellitus (T2DM), with acute COVID-19, and those with Long COVID/PASC symptoms. Results: We show that plasma samples from Long COVID/PASC still contain large anomalous (amyloid) deposits (microclots). We also show that these microclots in both acute COVID-19 and Long COVID/PASC plasma samples are resistant to fibrinolysis (compared to plasma from controls and T2DM), even after trypsinisation. After a second trypsinization, the persistent pellet deposits (microclots) were solubilized. We detected various inflammatory molecules that are substantially increased in both the supernatant and trapped in the solubilized pellet deposits of acute COVID-19 and Long COVID/PASC, versus the equivalent volume of fully digested fluid of the control samples and T2DM. Of particular interest was a substantial increase in α(2)-antiplasmin (α2AP), various fibrinogen chains, as well as Serum Amyloid A (SAA) that were trapped in the solubilized fibrinolytic-resistant pellet deposits. Conclusions: Clotting pathologies in both acute COVID-19 infection and in Long COVID/PASC might benefit from following a regime of continued anticlotting therapy to support the fibrinolytic system function.
MeSH term(s)	Adult ; Antifibrinolytic Agents/metabolism ; Blood Coagulation Factors/metabolism ; COVID-19/complications ; Disease Progression ; Female ; Humans ; Male ; Middle Aged ; SARS-CoV-2/pathogenicity ; Post-Acute COVID-19 Syndrome
Chemical Substances	Antifibrinolytic Agents ; Blood Coagulation Factors
Language	English
Publishing date	2021-08-23
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2093769-6
ISSN	1475-2840 ; 1475-2840
ISSN (online)	1475-2840
ISSN	1475-2840
DOI	10.1186/s12933-021-01359-7
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Erythrocyte, Platelet, Serum Ferritin, and P-Selectin Pathophysiology Implicated in Severe Hypercoagulation and Vascular Complications in COVID-19.

Venter, Chantelle / Bezuidenhout, Johannes Andries / Laubscher, Gert Jacobus / Lourens, Petrus Johannes / Steenkamp, Janami / Kell, Douglas B / Pretorius, Etheresia

International journal of molecular sciences

2020 Volume 21, Issue 21

Abstract: Progressive respiratory failure is seen as a major cause of death in severe acute respiratory syndrome coronavirus 2 (SARS-Cov-2)-induced infection. Relatively little is known about the associated morphologic and molecular changes in the circulation of ... ...

Abstract	Progressive respiratory failure is seen as a major cause of death in severe acute respiratory syndrome coronavirus 2 (SARS-Cov-2)-induced infection. Relatively little is known about the associated morphologic and molecular changes in the circulation of these patients. In particular, platelet and erythrocyte pathology might result in severe vascular issues, and the manifestations may include thrombotic complications. These thrombotic pathologies may be both extrapulmonary and intrapulmonary and may be central to respiratory failure. Previously, we reported the presence of amyloid microclots in the circulation of patients with coronavirus disease 2019 (COVID-19). Here, we investigate the presence of related circulating biomarkers, including C-reactive protein (CRP), serum ferritin, and P-selectin. These biomarkers are well-known to interact with, and cause pathology to, platelets and erythrocytes. We also study the structure of platelets and erythrocytes using fluorescence microscopy (using the markers PAC-1 and CD62PE) and scanning electron microscopy. Thromboelastography and viscometry were also used to study coagulation parameters and plasma viscosity. We conclude that structural pathologies found in platelets and erythrocytes, together with spontaneously formed amyloid microclots, may be central to vascular changes observed during COVID-19 progression, including thrombotic microangiopathy, diffuse intravascular coagulation, and large-vessel thrombosis, as well as ground-glass opacities in the lungs. Consequently, this clinical snapshot of COVID-19 strongly suggests that it is also a true vascular disease and considering it as such should form an essential part of a clinical treatment regime.
MeSH term(s)	Betacoronavirus/isolation & purification ; Blood Coagulation/physiology ; Blood Platelets/pathology ; Blood Platelets/virology ; COVID-19 ; Cardiovascular Diseases/blood ; Cardiovascular Diseases/pathology ; Cardiovascular Diseases/virology ; Coronavirus Infections/blood ; Coronavirus Infections/pathology ; Coronavirus Infections/virology ; Erythrocytes/pathology ; Erythrocytes/virology ; Female ; Ferritins/blood ; Humans ; Male ; Middle Aged ; P-Selectin/blood ; Pandemics ; Pneumonia, Viral/blood ; Pneumonia, Viral/pathology ; Pneumonia, Viral/virology ; SARS-CoV-2 ; Thrombosis/pathology ; Thrombosis/virology
Chemical Substances	P-Selectin ; Ferritins (9007-73-2)
Keywords	covid19
Language	English
Publishing date	2020-11-03
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2019364-6
ISSN	1422-0067 ; 1422-0067 ; 1661-6596
ISSN (online)	1422-0067
ISSN	1422-0067 ; 1661-6596
DOI	10.3390/ijms21218234
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article: Erythrocyte, Platelet, Serum Ferritin, and P-Selectin Pathophysiology Implicated in Severe Hypercoagulation and Vascular Complications in COVID-19

Venter, Chantelle Bezuidenhout Johannes Andries Laubscher Gert Jacobus Lourens Petrus Johannes Steenkamp Janami Kell Douglas B. / Pretorius, Etheresia

International Journal of Molecular Sciences

Abstract: Progressive respiratory failure is seen as a major cause of death in severe acute respiratory syndrome coronavirus 2 (SARS-Cov-2)-induced infection Relatively little is known about the associated morphologic and molecular changes in the circulation of ... ...

Abstract	Progressive respiratory failure is seen as a major cause of death in severe acute respiratory syndrome coronavirus 2 (SARS-Cov-2)-induced infection Relatively little is known about the associated morphologic and molecular changes in the circulation of these patients In particular, platelet and erythrocyte pathology might result in severe vascular issues, and the manifestations may include thrombotic complications These thrombotic pathologies may be both extrapulmonary and intrapulmonary and may be central to respiratory failure Previously, we reported the presence of amyloid microclots in the circulation of patients with coronavirus disease 2019 (COVID-19) Here, we investigate the presence of related circulating biomarkers, including C-reactive protein (CRP), serum ferritin, and P-selectin These biomarkers are well-known to interact with, and cause pathology to, platelets and erythrocytes We also study the structure of platelets and erythrocytes using fluorescence microscopy (using the markers PAC-1 and CD62PE) and scanning electron microscopy Thromboelastography and viscometry were also used to study coagulation parameters and plasma viscosity We conclude that structural pathologies found in platelets and erythrocytes, together with spontaneously formed amyloid microclots, may be central to vascular changes observed during COVID-19 progression, including thrombotic microangiopathy, diffuse intravascular coagulation, and large-vessel thrombosis, as well as ground-glass opacities in the lungs Consequently, this clinical snapshot of COVID-19 strongly suggests that it is also a true vascular disease and considering it as such should form an essential part of a clinical treatment regime
Keywords	covid19
Publisher	WHO
Document type	Article
Note	WHO #Covidence: #896573
Database	COVID19

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Article: Prevalence of readily detected amyloid blood clots in 'unclotted' Type 2 Diabetes Mellitus and COVID-19 plasma: a preliminary report

Pretorius, Etheresia / Venter, Chantelle / Laubscher, Gert Jacobus / Lourens, Petrus Johannes / Steenkamp, Janami / Kell, Douglas B

Cardiovasc Diabetol

Abstract: BACKGROUND: Type 2 Diabetes Mellitus (T2DM) is a well-known comorbidity to COVID-19 and coagulopathies are a common accompaniment to both T2DM and COVID-19. In addition, patients with COVID-19 are known to develop micro-clots within the lungs. The rapid ... ...

Abstract	BACKGROUND: Type 2 Diabetes Mellitus (T2DM) is a well-known comorbidity to COVID-19 and coagulopathies are a common accompaniment to both T2DM and COVID-19. In addition, patients with COVID-19 are known to develop micro-clots within the lungs. The rapid detection of COVID-19 uses genotypic testing for the presence of SARS-Cov-2 virus in nasopharyngeal swabs, but it can have a poor sensitivity. A rapid, host-based physiological test that indicated clotting severity and the extent of clotting pathologies in the individual who was infected or not would be highly desirable. METHODS: Platelet poor plasma (PPP) was collected and frozen. On the day of analysis, PPP samples were thawed and analysed. We show here that microclots can be detected in the native plasma of twenty COVID-19, as well as ten T2DM patients, without the addition of any clotting agent, and in particular that such clots are amyloid in nature as judged by a standard fluorogenic stain. Results were compared to ten healthy age-matched individuals. RESULTS: In COVID-19 plasma these microclots are significantly increased when compared to the levels in T2DM. CONCLUSIONS: This fluorogenic test may provide a rapid and convenient test with 100% sensitivity (P < 0.0001) and is consistent with the recognition that the early detection and prevention of such clotting can have an important role in therapy.
Keywords	covid19
Publisher	WHO
Document type	Article
Note	WHO #Covidence: #925887
Database	COVID19

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Article ; Online: Prevalence of amyloid blood clots in COVID-19 plasma

Pretorius, Etheresia / Venter, Chantelle / Laubscher, Gert J / Lourens, Petrus J / Steenkamp, Janami / Kell, Douglas B

medRxiv

Abstract: The rapid detection of COVID-19 uses genotypic testing for the presence of SARS-Cov-2 virus in nasopharyngeal swabs, but it can have a poor sensitivity. A rapid, host-based physiological test that indicated whether the individual was infected or not ... ...

Abstract	The rapid detection of COVID-19 uses genotypic testing for the presence of SARS-Cov-2 virus in nasopharyngeal swabs, but it can have a poor sensitivity. A rapid, host-based physiological test that indicated whether the individual was infected or not would be highly desirable. Coagulaopathies are a common accompaniment to COVID-19, especially micro-clots within the lungs. We show here that microclots can be detected in the native plasma of COVID-19 patient, and in particular that such clots are amyloid in nature as judged by a standard fluorogenic stain. This provides a rapid and convenient test (P<0.0001), and suggests that the early detection and prevention of such clotting could have an important role in therapy.
Keywords	covid19
Language	English
Publishing date	2020-07-29
Publisher	Cold Spring Harbor Laboratory Press
Document type	Article ; Online
DOI	10.1101/2020.07.28.20163543
Database	COVID19

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Article ; Online: Prevalence of Readily Detected Amyloid Blood Clots in ‘Unclotted’ COVID-19 Plasma

Pretorius, Etheresia / Venter, Chantelle / Laubscher, Gert Jacobus / Lourens, Petrus Johannes / Steenkamp, Janami / Kell, Douglas

SSRN Electronic Journal ; ISSN 1556-5068

2020

Keywords	covid19
Language	English
Publisher	Elsevier BV
Publishing country	us
Document type	Article ; Online
DOI	10.2139/ssrn.3670690
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Prevalence of amyloid blood clots in COVID-19 plasma

Pretorius, Etheresia / Venter, Chantelle / Laubscher, Gert J. / Lourens, Petrus J. / Steenkamp, Janami / Kell, Douglas B.

2020

Abstract: CITATION: Pretorius, E. et al. 2020. Prevalence of amyloid blood clots in COVID-19 plasma. medRxiv, doi:10.1101/2020.07.28.20163543. ... The original publication is available at https://www.medrxiv.org ... The rapid detection of COVID-19 uses genotypic ... ...

Abstract	CITATION: Pretorius, E. et al. 2020. Prevalence of amyloid blood clots in COVID-19 plasma. medRxiv, doi:10.1101/2020.07.28.20163543. The original publication is available at https://www.medrxiv.org The rapid detection of COVID-19 uses genotypic testing for the presence of SARS-Cov-2 virus in nasopharyngeal swabs, but it can have a poor sensitivity. A rapid, host-based physiological test that indicated whether the individual was infected or not would be highly desirable. Coagulaopathies are a common accompaniment to COVID-19, especially micro-clots within the lungs. We show here that microclots can be detected in the native plasma of COVID-19 patient, and in particular that such clots are amyloid in nature as judged by a standard fluorogenic stain. This provides a rapid and convenient test (P<0.0001), and suggests that the early detection and prevention of such clotting could have an important role in therapy. https://www.medrxiv.org/content/10.1101/2020.07.28.20163543v1 Pre-print
Keywords	COVID-19 (Disease) ; Blood -- Coagulation ; Blood plasma ; covid19
Language	English
Publisher	medRxiv
Publishing country	za
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: SARS-CoV-2 spike protein S1 induces fibrin(ogen) resistant to fibrinolysis: Implications for microclot formation in COVID-19

Grobbelaar, Lize M / Venter, Chantelle / Vlok, Mare / Ngoepe, Malebogo / Laubscher, Gert J / Lourens, Petrus J / Steenkamp, Janami / Kell, Douglas B / Pretorius, Etheresia

medRxiv

Abstract: Severe acute respiratory syndrome coronavirus 2 (SARS-Cov-2) -induced infection, the cause of coronavirus disease 2019 (COVID-19), is characterized by unprecedented clinical pathologies. One of the most important pathologies, is hypercoagulation and ... ...

Abstract	Severe acute respiratory syndrome coronavirus 2 (SARS-Cov-2) -induced infection, the cause of coronavirus disease 2019 (COVID-19), is characterized by unprecedented clinical pathologies. One of the most important pathologies, is hypercoagulation and microclots in the lungs of patients. Here we study the effect of isolated SARS-CoV-2 spike protein S1 subunit as potential inflammagen sui generis. Using scanning electron and fluorescence microscopy as well as mass spectrometry, we investigate the potential of this inflammagen to interact with platelets and fibrin(ogen) directly to cause blood hypercoagulation. Using platelet poor plasma (PPP), we show that spike protein may interfere with blood flow. Mass spectrometry also showed that when spike protein S1 is added to healthy PPP, it results in structural changes to β and γ fibrin(ogen), complement 3, and prothrombin. These proteins were substantially resistant to trypsinization, in the presence of spike protein S1. Here we suggest that, in part, the presence of spike protein in circulation may contribute to the hypercoagulation in COVID-19 positive patients and may cause substantial impairment of fibrinolysis. Such lytic impairment may result in the persistent large microclots we have noted here and previously in plasma samples of COVID-19 patients. This observation may have important clinical relevance in the treatment of hypercoagulability in COVID-19 patients.
Keywords	covid19
Language	English
Publishing date	2021-03-08
Publisher	Cold Spring Harbor Laboratory Press
Document type	Article ; Online
DOI	10.1101/2021.03.05.21252960
Database	COVID19

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