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      The effect of physiological levels of South African puff adder (Bitis arietans) snake venom on blood cells: an in vitro model

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          Abstract

          A significant burden of illness is caused globally by snakebites particularly by the puff adder, Bitis arietans. Presently there is no reliable and rapid method to confirm envenomation on blood chemistry; although coagulation parameters like prothrombin time, partial thromboplastin time, international normalized ratio and also serum electrolytes are tested. Here, we found that direct in vitro exposure of physiological relevant whole venom levels to human healthy blood (N = 32), caused significant physiological changes to platelet activity using a hematology analyzer, and measuring occlusion time, as well as lyses time, with the global thrombosis test (GTT). Disintegrated platelets were confirmed by scanning electron microscopy (SEM). We also confirmed the pathologic effects on erythrocytes (RBCs) (visible as eryptotic RBCs), by looking at both light microscopy and SEM. Thromboelastography showed that no clot formation in whole blood could be induced after addition of whole venom. We propose further clinical studies to investigate the use of light microscopy smears and hematology analyzer results immediately after envenomation, as a possible first-stage of clinical confirmation of envenomation.

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          Changes in red blood cell membrane structure in type 2 diabetes: a scanning electron and atomic force microscopy study

          Red blood cells (RBCs) are highly deformable and possess a robust membrane that can withstand shear force. Previous research showed that in diabetic patients, there is a changed RBC ultrastructure, where these cells are elongated and twist around spontaneously formed fibrin fibers. These changes may impact erythrocyte function. Ultrastructural analysis of RBCs in inflammatory and degenerative diseases can no longer be ignored and should form a fundamental research tool in clinical studies. Consequently, we investigated the membrane roughness and ultrastructural changes in type 2 diabetes. Atomic force microscopy (AFM) was used to study membrane roughness and we correlate this with scanning electron microscopy (SEM) to compare results of both the techniques with the RBCs of healthy individuals. We show that the combined AFM and SEM analyses of RBCs give valuable information about the disease status of patients with diabetes. Effectiveness of treatment regimes on the integrity, cell shape and roughness of RBCs may be tracked, as this cell’s health status is crucial to the overall wellness of the diabetic patient.
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            The simultaneous occurrence of both hypercoagulability and hypofibrinolysis in blood and serum during systemic inflammation, and the roles of iron and fibrin(ogen).

            Although the two phenomena are usually studied separately, we summarise a considerable body of literature to the effect that a great many diseases involve (or are accompanied by) both an increased tendency for blood to clot (hypercoagulability) and the resistance of the clots so formed (hypofibrinolysis) to the typical, 'healthy' or physiological lysis. We concentrate here on the terminal stages of fibrin formation from fibrinogen, as catalysed by thrombin. Hypercoagulability goes hand in hand with inflammation, and is strongly influenced by the fibrinogen concentration (and vice versa); this can be mediated via interleukin-6. Poorly liganded iron is a significant feature of inflammatory diseases, and hypofibrinolysis may change as a result of changes in the structure and morphology of the clot, which may be mimicked in vitro, and may be caused in vivo, by the presence of unliganded iron interacting with fibrin(ogen) during clot formation. Many of these phenomena are probably caused by electrostatic changes in the iron-fibrinogen system, though hydroxyl radical (OH˙) formation can also contribute under both acute and (more especially) chronic conditions. Many substances are known to affect the nature of fibrin polymerised from fibrinogen, such that this might be seen as a kind of bellwether for human or plasma health. Overall, our analysis demonstrates the commonalities underpinning a variety of pathologies as seen in both hypercoagulability and hypofibrinolysis, and offers opportunities for both diagnostics and therapies.
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              Impaired endogenous thrombolysis in acute coronary syndrome patients predicts cardiovascular death and nonfatal myocardial infarction.

              Our objective was to assess endogenous thrombolytic activity in acute coronary syndrome (ACS) patients and relate this to their likelihood of future adverse cardiovascular events. Spontaneous lysis of platelet-rich thrombi is an important defense mechanism against lasting occlusion. Despite convincing evidence for the role of endogenous fibrinolysis in ACS, the prognostic value of plasma fibrinolytic markers in assessing risk is limited. We employed a novel global test which, in addition to platelet reactivity, allows assessment of endogenous thrombolytic activity to identify ACS patients who remain at risk of cardiovascular events. We used the global thrombosis test (GTT) to assess thrombotic and thrombolytic status in 300 ACS patients receiving dual-antiplatelet therapy. The test assesses the time required to form an occlusive thrombus, the occlusion time (OT), and the time to lyse this, the lysis time (LT). The end point of the study at 12 months' follow-up was the composite of death, nonfatal myocardial infarction, or stroke. The OT and LT were both prolonged in ACS patients compared with normal volunteers (p or =3,000 s occurred in 23% of ACS patients versus none of the normal subjects and was a significant and independent predictor of cardiovascular death and nonfatal myocardial infarction in a multivariate model adjusted for cardiovascular risk factors. LT > or =3,000 s was the optimal cutoff value for predicting 12-month major adverse cardiovascular events (hazard ratio [HR]: 2.52, 95% confidence interval: 1.34 to 4.71, p = 0.004) and cardiovascular death (HR: 4.2, 95% confidence interval: 1.13 to 15.62, p = 0.033). HR increased further as LT increased. No association was found between OT and the risk of major adverse cardiovascular events. Assessment of endogenous thrombolytic status based on the lysis of platelet-rich thrombi from native blood using the point-of-care GTT can identify ACS patients at risk of future cardiac events. Copyright 2010 American College of Cardiology Foundation. Published by Elsevier Inc. All rights reserved.
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                Author and article information

                Journal
                Sci Rep
                Sci Rep
                Scientific Reports
                Nature Publishing Group
                2045-2322
                24 October 2016
                2016
                : 6
                : 35988
                Affiliations
                [1 ]Department of Physiology, University of Pretoria , South Africa
                Author notes
                Article
                srep35988
                10.1038/srep35988
                5075924
                27775063
                83b0ea2f-cdee-4c61-accb-81d2aa466689
                Copyright © 2016, The Author(s)

                This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

                History
                : 02 August 2016
                : 10 October 2016
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