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Introduction: Oxidative stress plays a critical role in the progression of diabetes, arthritis, cancer, eryptosis, cardiovascular disease, and thrombosis. Currently, antioxidants from natural sources are in high demand due to their beneficial role in the management of said diseases. Aim: The purpose of the study was to evaluate the protective effect of sorghum protein buffer extract (SBE) on sodium nitrite-induced oxidative stress and thrombosis. Materials and methods: Protein characterization of SBE was done using SDS-PAGE. Oxidative stress in RBC was induced using sodium nitrite (NaNO2) and the key stress markers such as lipid peroxidation (LPO), protein carbonyl content (PCC), and the level of antioxidant enzymes (SOD and CAT) were measured. The anticoagulant effect of SBE was identified by employing in-vitro plasma recalcification time, activated partial thromboplastin time (APTT), prothrombin time (PT), and in-vivo mouse tail bleeding time. SBE antiplatelet activity was examined using agonist adenosine diphosphate (ADP) and epinephrine-induced platelet aggregation. Non-toxic property of SBE was identified using in-vitro direct haemolytic, haemorrhagic, and edema forming activities using experimental mice. Results: SBE revealed similar protein banding pattern under both reduced and non-reduced conditions on SDS-PAGE. Interestingly, SBE normalized the level of LPO, PCC, SOD, and CAT in stress-induced RBCs. Furthermore, SBE showed anticoagulant effect in platelet rich plasma by enhancing the clotting time from the control 250 s to 610 s and bleeding time from the control 200 s to more than 500 s (p Conclusions: This study demonstrates for the first time the anticoagulant, antiplatelet, and antioxidant properties of SBE. Thus, the observed results validate consumption of sorghum as good for health and well-being.

To understand the RBC protecting efficiency and antiplatelet activity of methanolic extract of Caesalpinia crista coat (MECCC). RBC-protecting activity of MECCC was evaluated using assays, such as DPPH, level of lipid peroxidation, protein carbonyl content, superoxide dismutase and catalase as a marker of oxidative stress whereas, platelet aggregation inhibition was performed using human platelet-rich plasma (PRP). MECCC showed about 76% of DPPH-scavenging activity, with an IC50 value of 71.89 μg/ml. The MECCC reduced the level of lipid peroxidation and protein carboxylation in RBC caused by NaNO2 in a dose-dependent manner. In addition, MECCC normalized the levels of superoxide dismutase (SOD) and catalase (CAT) in oxidative stress-induced RBC in a dose-dependent manner. This suggested the protective effect of MECCC on RBC against oxidative stress. Furthermore, MECCC also exhibited mild antiplatelet activity by inhibiting both ADP and epinephrine agonists that induced platelet aggregation. The noticed inhibition percentage was found to be 28 and 23%, respectively at the concentration of 150 μg. Interestingly, MECCC did not hydrolyse the RBC suggesting its nontoxic properties. MECCC possesses protective effect of RBC against NaNO2 (10 mmol/l) induce oxidative stress and inhibits platelet aggregation.

Suicidal erythrocyte death, or eryptosis, is the key event in eliciting anemia in numerous pathological conditions, including diabetes, chronic kidney disease, cancer, sepsis, etc. Oxidative stress is an important trigger in the acceleration of erythrocyte loss via eryptosis and an underlying mechanism of anemia emergence in the above pathologies. Therefore, there is an increasing demand for identification of antioxidants and anti-eryptotic agents for the management of stress-related ailments. Here, we demonstrated the antioxidant and anti-eryptotic properties of the tamarind seed coat ethanol extract (TSCEE) against 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH)-induced oxidative stress and eryptosis. The presence of probable secondary metabolites in the TSCEE extract was investigated by RP-HPLC. Active groups present in the TSCEE were studied by the Fourier-transform infrared spectroscopy. Cyclic voltammetric studies confirmed the antioxidant potential of TSCEE. The protective effect of TSCEE on red blood cells was confirmed by assessing various eryptotic markers, such as reactive oxygen species generation, intracellular calcium levels, and phosphatidylserine exposure. TSCEE reduced lipid peroxidation and protein carbonyl content and restored the levels of glutathione, antioxidant enzymes, and enzymes involved in glutathione replenishment. In conclusion, TSCEE was found to exhibit multiple therapeutic properties, which makes it a promising agent for treating oxidative stress-induced eryptosis and subsequent anemia in various pathologies.

The current study deals with the purification and characterization of non-enzymatic glycoprotein (NEGp) from flax seed buffer extract. Sephadex G-100 and DEAE-A25 column chromatography techniques were employed to isolate NEGp. NEGp showed single sharp band at 29 kDa region on 10% SDS-PAGE, and under reduced and non-reduced conditions revealed its monomeric nature. Besides, NEGp taken up the PAS stain at 29 kDa region reveals the presence of carbohydrate moiety. Purity of NEGp was adjudged by RP-HPLC, as it revealed a single sharp peak at the retention time of 3.4 min. The exact molecular mass of NEGp was found to be 26 kDa which was confirmed by MALDI-TOF. Circular di-chromism spectra of NEGp showed 12.0% α-helix, 24.3% α-helix turn and 63.7% random coils without beta pleated sheets. NEGp was found to exhibit anticoagulant activity by extending clotting time of both platelet rich plasma and platelet poor plasma from control 240 s to 1800 s and 280 s to 2100 s respectively at the concentration of 8 μg. NEGp inhibited the agonists such as ADP, epinephrine and arachidonic acid induced platelet aggregation in washed platelets. The percentage of inhibition was found to be 70%, 80% and 60% respectively. While, it did not interfere in thrombin, PAF and collagen induced platelet aggregation. NEGp did not hydrolyse RBC membrane, devoid of haemorrhagic and edema inducing properties in experimental mice.

Background: Flax seeds possess long array of medicinal qualities as it found to show beneficial effects on cardiovascular, hypertension, diabetes, cancer, inflammatory and autoimmune disorders. While, the therapeutic role of flax seed proteins on thrombotic disorder is least explored. Objective: The current study aims to identify the fibrin clot-dissolving, anticoagulant and antiplatelet properties of flaxseed buffer extract (FSBE) using Platelet Rich Plasma (PRP) and Platelet Poor Plasma (PPP). Materials and Methods: Flax Seed Buffer Extract (FSBE) proteins were characterized upon subjected to SDS-PAGE. The presence of cysteine protease in the extract was identified using colorimeter and zymography experiments. Anticoagulant activity was established using invitro recalcification time. While, the fibrinogenolytic, fibrinolytic and human plasma proteins degradation activities were proved using SDS-PAGE. Furthermore, antiplatelet activity was confirmed using Chronology dual channel whole blood/optical lumi aggregation system (Model-700). Toxicity studies were assayed using RBC cells and albino mice. Results: FSBE showed divergent protein banding pattern from the molecular mass ranging 200 kDa-14 kDa under both reduced and non-reduced conditions. The FSBE showed proteolytic activity as it hydrolyzed casein and gelatin with the specific activity 0.180 and 0.201 units/mg/min respectively. The proteolytic activity of FSBE was completely abolished only by IAA but PMSF, 1, 10 Phenanthroline and EDTA did not, revealing the presence of cysteine protease in the extract. FSBE showed anticoagulant activity as it enhanced the clotting time from control 220s to 320s. Furthermore, FSBE showed antiplatelet activity by inhibiting ADP and Epinephrine induced platelet aggregation and the observed aggregation inhibition was found to be 63.6% and 16.3% respectively. In addition, it hydrolyzed specifically Aα and Bβ chains of human fibrinogen and all the chains of human fibrin clot. The fibrinogenolytic activity was inhibited by IAA but PMSF, EDTA, 1, 10 Phenanthroline did not, suggesting the role of cysteine proteases. Interestingly, FSBE did not cause edema and hemorrhage in the experimental mice and did not hydrolyze RBC cells suggesting its nontoxic properties. Abbreviations used: FSBE: Flaxseed buffer extract; PMSF: Phenylmethylsulfonyl fluoride; IAA: Iodoacetic acid; EDTA: Ethylenediaminetetraacetic acid; PRP: Platelet-rich plasma; PPP: Platelet-poor plasma; INR: International normalized ratio; MED: Minimum edema dose; MHD: Minimum hemorrhagic dose; APTT: Activated partial thromboplastin time; PT: Prothrombin time; HMWK: High-molecular-weight kininogen.