Viscoelasticity as a measurement of clot structure in poorly controlled type 2 diabetes patients : towards a precision and personalized medicine approach

OBJECTIVES : Type 2 diabetes patients (T2D) have a considerably higher cardiovascular risk, which is closely associated with systemic inflammation, and an accompanying pathologic coagulation system. Due to the complexity of the diabetic profile, we suggest that we need to look at each patient individually and particularly at his or her clotting profile; as the healthiness of the coagulation system gives us an indication of the success of clinical intervention. RESULTS : T2D coagulability varied markedly, although there were no clear difference in medication use and the standards of HbA1c levels. RESEARCH DESIGN AND METHODS : Our sample consisted of 90 poorly controlled T2D and 71 healthy individuals. We investigated the medication use and standards of HbA1c levels of T2D and we used thromboelastography (TEG) and scanning electron microscopy (SEM) to study their clot formation. CONCLUSION : The latest NIH guidelines suggest that clinical medicine should focus on precision medicine, and the current broad understanding is that precision medicine may in future, provide personalized targets for preventative and therapeutic interventions. Here we suggest a practical example where TEG can be used as an easily accessible point-of-care tool to establish a comprehensive clotting profile analysis for T2D patients ; and additionally may provide valuable information that may be used in the envisaged precision medicine approach. Only by closely following each individual patient’s progress and healthiness and thereby managing systemic inflammation, will we be able to reduce this pandemic.National Research Foundation (NRF) of South Africa (91548: Competitive Program) and Medical Research Council (MRC) of South Africa (Self-Initiated Research Program: A0X331). Grant holder: E Pretorius.http://www.impactjournals.com/oncotargethb2016Physiolog

Effects of IL-1β, IL-6 and IL-8 on erythrocytes, platelets and clot viscoelasticity

Complex interactions exist between cytokines, and the interleukin family plays a fundamental role in inflammation. Particularly circulating IL-1β, IL-6 and IL-8 are unregulated in systemic and chronic inflammatory conditions. Hypercoagulability is an important hallmark of inflammation, and these cytokines are critically involved in abnormal clot formation, erythrocyte pathology and platelet hyper-activation, and these three cytokines have known receptors on platelets. Although these cytokines are always unregulated in inflammation, we do not know how the individual cytokines act upon the structure of erythrocytes and platelets, and which of the viscoelastic clot parameters are changed. Here we study the effects of IL-1β, IL-6 and IL-8 at low physiological levels, representative of chronic inflammation, by using scanning electron microscopy and thromboelastography. All three interleukins caused the viscoelastic properties to display an increased hypercoagulability of whole blood and pathology of both erythrocytes and platelets. The most pronounced changes were noted where all three cytokines caused platelet hyper-activation and spreading. Erythrocyte structure was notably affected in the presence of IL-8, where the morphological changes resembled that typically seen in eryptosis (programmed cell death). We suggest that erythrocytes and platelets are particularly sensitive to cytokine presence, and that they are excellent health indicators.National Research Foundation (NRF) of South Africa (91548: Competitive Program) and Medical Research Council (MRC) of South Africa (Self-Initiated Research Program: A0× 331). Grant holder: E Pretorius.www.nature.com/scientificreports/am2016Physiolog

Pathophysiological changes in erythrocytes contributing to complications of inflammation and coagulation in COVID-19

Higher thrombotic burden in the acute phase of COVID-19 relies on a complex interplay between pro-inflammatory cytokine/chemokine release, increased endothelial dysfunction/ damage, and potential sepsis-induced coagulopathy development in severe cases, all promoting coagulation activation. Plasma levels of cytokines and chemokines are known to be increased in COVID-19 however, are much higher in severe infections. Increased levels of IL1β, IL-6, and IL-8 are known to play an important role in both acute and chronic inflammation, resulting in pathological clotting. However, little has been published on the effects of these interleukins on red blood cells (RBCs). Evidence shows that cytokines have a negative effect on the RBCs ultrastructure and introduce signs of eryptosis. Eryptosis can be described as a form of suicidal death of RBCs characterized by distinct findings of cell shrinkage, membrane blebbing, activation of proteases, and phosphatidylserine exposure at the outer membrane leaflet. Red blood cells from COVID-19 patients had increased levels of glycolytic intermediates, accompanied by oxidation and fragmentation of ankyrin, spectrin beta, and the N-terminal cytosolic domain of band 3 (AE1). Significantly altered lipid metabolism was also observed, in particular, short- and medium-chain saturated fatty acids, acyl-carnitines, and sphingolipids. Emerging research suggests that RBCs may contribute to a precision medicine approach to sepsis and have diagnostic value in monitoring complement dysregulation in COVID-19-sepsis and non-COVID sepsis as research indicates that complement activation products and viral antigens are present on RBCs in patients with COVID-19.https://www.frontiersin.org/journals/physiologydm2022AnatomyPhysiolog

The dormant blood microbiome in chronic, inflammatory diseases

Blood in healthy organisms is seen as a ‘sterile’ environment: it lacks proliferating microbes. Dormant or not-immediately-culturable forms are not absent, however, as intracellular dormancy is well established. We highlight here that a great many pathogens can survive in blood and inside erythrocytes. ‘Non-culturability’, reflected by discrepancies between plate counts and total counts, is commonplace in environmental microbiology. It is overcome by improved culturing methods, and we asked how common this would be in blood. A number of recent, sequence-based and ultramicroscopic studies have uncovered an authentic blood microbiome in a number of non-communicable diseases. The chief origin of these microbes is the gut microbiome (especially when it shifts composition to a pathogenic state, known as ‘dysbiosis’). Another source is microbes translocated from the oral cavity. ‘Dysbiosis’ is also used to describe translocation of cells into blood or other tissues. To avoid ambiguity, we here use the term ‘atopobiosis’ for microbes that appear in places other than their normal location. Atopobiosis may contribute to the dynamics of a variety of inflammatory diseases. Overall, it seems that many more chronic, non-communicable, inflammatory diseases may have a microbial component than are presently considered, and may be treatable using bactericidal antibiotics or vaccines.Biotechnology and Biological Sciences Research Council (grant BB/L025752/1) as well as the National Research Foundation (NRF) of South Africa.http://femsre.oxfordjournals.orghb201

Atypical erythrocytes and platelets in a patient with a pro-thrombin mutation

Prothrombin mutation G20210A, anti-phospholipid syndrome as well as iron overload has previously been shown to cause thrombotic events. The main reason for this is the involvement of these anomalies in causing hypercoagulability of the coagulation system, which frequently leads to venous and arterial thrombotic events. We report the case of a 37-year-old white female with prothrombin mutation G20210A, anti-phospholipid syndrome, as well as an increased serum ferritin level, who experienced two transient ischemic attacks and suffers from regular amaurosis fugax. We present an ultrastructural depiction of erythrocytes, platelets, and the fibrin network, to explain the clinical manifestations of the thrombotic state seen in this patient.http://informahealthcare.com/plt2015-06-30hb201

High ferritin levels have major effects on the morphology of erythrocytes in Alzheimer's disease

Introduction: Unliganded iron both contributes to the pathology of Alzheimer's disease (AD) and also changes the morphology of erythrocytes (RBCs). We tested the hypothesis that these two facts might be linked, i.e., that the RBCs of AD individuals have a variant morphology, that might have diagnostic or prognostic value. Methods: We included a literature survey of AD and its relationships to the vascular system, followed by a laboratory study. Four different microscopy techniques were used and results statistically compared to analyze trends between high and normal serum ferritin (SF) AD individuals. Results: Light and scanning electron microscopies showed little difference between the morphologies of RBCs taken from healthy individuals and from normal SF AD individuals. By contrast, there were substantial changes in the morphology of RBCs taken from high SF AD individuals. These differences were also observed using confocal microscopy and as a significantly greater membrane stiffness (measured using force-distance curves). Conclusion: We argue that high ferritin levels may contribute to an accelerated pathology in AD. Our findings reinforce the importance of (unliganded) iron in AD, and suggest the possibility both of an early diagnosis and some means of treating or slowing down the progress of this disease

Editorial : Pathological changes in erythrocytes during inflammation and infection

No abstract available.https://www.frontiersin.org/journals/physiologydm2022Physiolog

Profound Morphological Changes in the Erythrocytes and Fibrin Networks of Patients with Hemochromatosis or with Hyperferritinemia, and Their Normalization by Iron Chelators and Other Agents

It is well-known that individuals with increased iron levels are more prone to thrombotic diseases, mainly due to the presence of unliganded iron, and thereby the increased production of hydroxyl radicals. It is also known that erythrocytes (RBCs) may play an important role during thrombotic events. Therefore the purpose of the current study was to assess whether RBCs had an altered morphology in individuals with hereditary hemochromatosis (HH), as well as some who displayed hyperferritinemia (HF). Using scanning electron microscopy, we also assessed means by which the RBC and fibrin morphology might be normalized. An important objective was to test the hypothesis that the altered RBC morphology was due to the presence of excess unliganded iron by removing it through chelation. Very striking differences were observed, in that the erythrocytes from HH and HF individuals were distorted and had a much greater axial ratio compared to that accompanying the discoid appearance seen in the normal samples. The response to thrombin, and the appearance of a platelet-rich plasma smear, were also markedly different. These differences could largely be reversed by the iron chelator desferal and to some degree by the iron chelator clioquinol, or by the free radical trapping agents salicylate or selenite (that may themselves also be iron chelators). These findings are consistent with the view that the aberrant morphology of the HH and HF erythrocytes is caused, at least in part, by unliganded (‘free’) iron, whether derived directly via raised ferritin levels or otherwise, and that lowering it or affecting the consequences of its action may be of therapeutic benefit. The findings also bear on the question of the extent to which accepting blood donations from HH individuals may be desirable or otherwise

Ultrastructural, confocal and viscoelastic characteristics of whole blood and plasma after exposure to cadmium and chromium alone and in combination : an ex vivo study

BACKGROUND/AIMS : Heavy metal pollution is increasing in the environment, contaminating water, food and air supplies. This can be linked to many anthropogenic activities. Heavy metals are absorbed through the skin, inhalation and/or orally. Irrespective of the manner of heavy metal entry in the body, the blood circulatory system is potentially the first to be affected following exposure and adverse effects on blood coagulation can lead to associated thrombotic disease. Although the plasma levels and the effects of cadmium (Cd) and chromium (Cr) on erythrocytes and lymphocytes have been described, the environmental exposure to heavy metals are not limited to a single metal and often involves metal mixtures, with each metal having different rates of absorption, different cellular, tissue, and organ targets. Therefore the aim of this study is to investigate the effects of the heavy metals Cd and Cr alone and whether Cr synergistically increases the effect of Cd on physiological important processes such as blood coagulation. METHODS : Human blood was exposed to the heavy metals ex vivo, and thereafter morphological analysis was performed with scanning electron- and confocal laser scanning microscopy (CLSM) in conjunction with thromboelastography®. RESULTS : The erythrocytes, platelets and fibrin networks presented with ultrastructural changes, including varied erythrocytes morphologies, activated platelets and significantly thicker fibrin fibres in the metal-exposed groups. CLSM analysis revealed the presence of phosphatidylserine on the outer surface of the membranes of the spherocytic erythrocytes exposed to Cd and Cr alone and in combination. The viscoelastic analysis revealed only a trend that indicates that clots that will form after heavy metal exposure, will likely be fragile and unstable especially for Cd and Cr in combination. CONCLUSION : This study identified the blood as an important target system of Cd and Cr toxicity.The National Research Foundation (NRF) (Grant number: 92768).am2017AnatomyPhysiolog

The effect of systemic lipopolysaccharides on the cardiovascular system in Sprague-Dawley rats using Manuka honey as a treatment

BACKGROUND : It has been said that there is a probable “head-to-heart” link in resulting in the development of Alzheimer’s disease (AD). This is because cardiovascular (CV) and neurodegenerative diseases such as AD share many risk factors. Researchers have theorised that a significant contributor to the neurological and CV complications in AD may be due to exposure to the lipopolysaccharide (LPS) endotoxin that results in a low-grade systemic inflammatory immune response. Systemic inflammation impacts both the CV and neurological systems in harmful ways. The elderly is more vulnerable to exposure since they are increasingly likely to suffer from tooth loss or gastrointestinal tract impairments resulting in LPS introduction into the CV system. AIM : The aim of the study was to investigate the effect of low physiological levels of systemic LPS exposure on the CV system, of male Sprague-Dawley rats over a ten-day period. Manuka honey was introduced as a possible mopping agent for the effects of LPS on the CV system. STUDY DESIGN : Twenty-four 10-week-old male Sprague-Dawley rats (average weight: 250-300 g) were randomly divided into four groups. The groups included rats exposed to Phosphate buffer-saline, phosphate buffered-saline and Manuka honey, LPS only as well as LPS and Manuka honey. The animals were housed for 19 days, in which the animals were given daily subcutaneous injections. The Manuka honey administered to the control and experimental groups received the Manuka honey solution through oral gavage starting on the 11th day for a seven-day period. METHODS : In this study we examined the CV tissue of the animals through light microscopy and transmission electron microscopy. The total cholesterol levels were also measured by using the QuickDetectTM total cholesterol (Rat) ELISA assay. Results: Myocardial LPS exposure resulted in cardiac myofibril damage and mitochondrial cristae destruction. The results indicated that Manuka honey may have alleviated some of this damage. However, the treatment failed to prevent mitochondrial cristae destruction. Lipopolysaccharide exposure to aortic tissue resulted in cellular abnormalities, elastic fragmentation and collagen deposition. Manuka honey administration resulted in the reduction of cellular abnormalities and elastin fragmentation in the aorta however, the treatment induced collagen depletion. CONCLUSION : The aorta and cardiac muscle are vulnerable to the effects of low levels of systemic LPS exposure. Exposure resulted in elevation in cardiac and aortic cellular and elastin destruction that may possibly conclude in CV disease long term. In addition, Manuka honey serves as an insufficient remedy to counteract the harmful effects of LPS on the CV system.National research fundPhysiolog