Department Of Zoology

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    Platelet-derived microvesicles activate human platelets via intracellular calcium mediated reactive oxygen species release
    (Academic Press Inc., 2022-08-28T00:00:00) Yadav, Pooja; Beura, Samir Kumar; Panigrahi, Abhishek Ramachandra; Bhardwaj, Taniya; Giri, Rajanish; Singh, Sunil Kumar
    Platelet-derived microvesicles (PMVs) are the most abundant microvesicles in circulation, originating from blood platelets via membrane blebbing. PMVs act as biological cargo carrying key molecules from platelets, including immunomodulatory molecules, growth factors, clotting molecules, and miRNAs that can regulate recipient cellular functions. Formation and release of PMVs play an essential role in the pathophysiology of vascular diseases such as hemostasis, inflammation, and thrombosis. Platelet activation is considered the critical event in thrombosis, and a growing number of evidence suggests that oxidative stress-mediated signaling plays a significant role in platelet activation. Ca2+ is a notable player in the generation of ROS in platelets. Reports have established that microvesicles exhibit dual nature in redox mechanisms as they possess both pro-oxidant and antioxidant machinery. However, the impact of PMVs and their ROS machinery on platelets is still a limited explored area. Here, we have demonstrated that PMVs mediate platelet activation via intracellular ROS generation. PMVs interacted with platelets and induced calcium-mediated intracellular ROS production via NADPH oxidase (NOX), leading to platelet activation. Our findings will open up new insights into the tangible relationship of PMVs with platelets and will further contribute to the therapeutic aspects of PMVs in vascular injury and tissue remodeling. � 2022
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    Mesoporous nanosilica: A thromboprotective nanomaterial for biomedical applications
    (Elsevier Ltd, 2022-06-17T00:00:00) Singh, Priti; Srivastava, Sameer; Singh, Sunil Kumar
    Nanosilica is widely employed in various biomedical applications because of their tailorable physiochemical properties and excellent biocompatibility. In the present study, we have evaluated interaction of nanosilica with important coagulation components, such as platelets, a highly sensitive cell found in the blood, and coagulation proteins. Mesoporous silica nanoparticles (MSNs) were prepared using sol-gel process and characterized by FESEM and TEM to find out the size and shape of the particles. Different platelet functional parameters including platelet adhesion, aggregation, activation, secretion, clot formation and clot retraction-based studies have been carried out to investigate the impact of synthesized nanosilica on the blood coagulation system. Besides, ROS generation and increase in intracellular calcium was also monitored as they play a pivotal role in regulating platelet functions. The complete detailed study revealed that MSNs neither has stimulatory action towards platelets nor do they show any effective interaction with coagulation proteins. � 2022 Elsevier Ltd
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    Role of Neurons and Glia Cells in Wound Healing as a Novel Perspective Considering Platelet as a Conventional Player
    (Springer, 2021-10-11T00:00:00) Beura, Samir K.; Panigrahi, Abhishek R.; Yadav, Pooja; Agrawal, Siwani; Singh, Sunil K.
    Wound healing is a complex physiological process in which the damaged or injured tissue is replaced or regenerated by new cells or existing cells respectively in their synthesized and secreted matrices. Several cells modulate the process of wound healing including macrophages, fibroblasts, and keratinocytes. Apart from these cells, platelet has been considered as a major cellular fragment to be involved in wound healing at several stages by secreting its granular contents including growth factors, thus resulting in coagulation, inflammation, and angiogenesis. A distant cell, which is gaining significant attention nowadays due to its resemblance with platelet in several aspects, is the neuron. Not only neurons but also glia cells are also confirmed to regulate wound healing at different stages in an orchestrated manner. Furthermore, these neurons and glia cells mediate wound healing inducing tissue repair and regeneration apart from hemostasis, angiogenesis, and inflammation by secreting various growth factors, coagulation molecules, immunomodulatory molecules as well as neurohormones, neuropeptides, and neurotrophins. Therefore, in wound healing platelets, neurons and glia cells not only contribute to tissue repair but are also responsible for establishing the wound microenvironment, thus affecting the proliferation of immune cells, fibroblast, and keratinocytes. Here in this review, we will enlighten the physiological roles of neurons and glia cells in coordination with platelets to understand various cellular and molecular mechanism in brain injury and associated neurocognitive impairments. � 2021, The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.
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    Pseudomonas aeruginosa quorum sensing molecule N-3-oxo-dodecanoyl-Lhomoserine lactone activates human platelets through intracellular calciummediated ROS generation
    (Elsevier, 2018) Yadav, V. K.; Singh, P. K.; Kalia, M.; Sharma, D.; Singh, S. K.; Agarwal, V.
    Pseudomonas aeruginosa, an opportunistic pathogen release N-3-oxo-dodecanoyl-l-homoserine lactone (3-oxo-C12HSL) and N-butyryl-l-homoserine lactone (C4-HSL) quorum sensing (QS) molecules to regulate various virulence factors responsible for infection in the host. 3-oxo-C12 HSL not only regulates the bacterial gene expression but also modulates the host cell system. Thus, it is pertinent to evaluate the effect of these QS molecules on blood platelets which is responsible for the maintenance of hemostasis and thrombus formation. Here, in the present study, we showed that 3-oxo-C12 HSL activates platelets in a dose-dependent manner and induces intracellular calcium-mediated reactive oxygen species (ROS) release, whereas no such effect was observed with C4-HSL. 3-oxo-C12 HSL stimulated ROS release was mediated by NADPH oxidase. Results confirmed the involvement of phospholipase C (PLC) and IP3 receptor behind intracellular calcium-mediated ROS generation. The impact of 3-oxo-C12 HSL on platelet activation suggests that it could interfere and alter the normal function of platelet in individuals infected with P. aeruginosa.
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    Development of lactadherin based electrochemical biosensor for the detection of platelet microvesicles
    (Central University of Punjab, 2018) Santra, Sneha; Singh, Sunil
    Platelet derived microvesicles (PMVs) are the most abundant membrane vesicles in the blood having a potent pro-inflammatory effect, promote coagulation and affect vascular function which are involved in the pathogenesis of cardiovascular disease including diabetes, thrombosis, and coronary artery diseases. Therefore, it is pertinent to detect PMVs level in blood of an individual which have prognostic potential for cardiovascular diseases. As per literature available, lactadherin is a small (53-66 kDa) multifunctional glycoprotein which plays an important role in the clearance of microvesicles. In the present study, lactadherin based electrochemical biosensor for the detection of PMVs was explored. Polythionine film (as a good electron mediator) was electrochemically deposited on ITO-coated glass through electrochemical process involving cyclic voltammetry (CV) and chronoamperometry. Electrochemically deposited electrode provides ideal adsorbing platform for immobilization of RGDS peptide sequences (Lactadherine binding motif) having binding affinity against active conformation of integrin on PMVs surface. CV and Diffusion Pulse Voltammetry (DPV) measurements showed gradual decrease of v current with the subsequent adsorption of microvesicles poor plasma (MPP), microvesicles rich plasma (MRP) and activated platelets. Decrease of current clearly depicts the presence of microvesicles in blood plasma. Our developed fabricated electrode can have a promising potential for its efficient application in clinical testing of various pathological conditions.