Potential applications of Fe3+-activated Sr9Al6O18 nanophosphors for fingerprint detection, oxidative stress, and thrombosis treatment.

This study reports on the synthesis of Fe3+-activated Sr9Al6O18 nanophosphors (SAO:Fe NPs) using a simple solution combustion process, which emits a pale green light and possesses excellent fluorescence properties. An in-situ powder dusting method was utilized to extract unique ridge features of latent fingerprints (LFPs) on various surfaces using ultra-violet 254 nm excitation. The results showed that SAO:Fe NPs possess high contrast, high sensitivity, and no background interference, enabling the observation of LFPs for longer periods. Poroscopy, which is the examination of sweat pores on the skin's papillary ridges, is important in the identification process, and the YOLOv8x program based on deep convolutional neural networks was used to study the features visible in FPs. The potential of SAO:Fe NPs to ameliorate oxidative stress and thrombosis was analyzed. The results showed that SAO:Fe NPs have antioxidant properties by scavenging 2,2-diphenylpicrylhydrazyl (DPPH) and normalized the stress markers in NaNO2-induced oxidative stress in Red Blood Cells (RBC). In addition, SAO:Fe inhibited platelet aggregation induced by adenosine diphosphate (ADP). Therefore, SAO:Fe NPs may have potential applications in advanced cardiology and forensic sciences. Overall, this study highlights the synthesis and potential applications of SAO:Fe NPs, which can enhance the sensitivity and specificity of fingerprint detection and provide insights into developing novel treatments for oxidative stress and thrombosis.

Anticoagulant and antiplatelet activities of novel serine protease purified from seeds of Cucumis maderaspatensis.

In this study, we report the biochemical characterization of a novel serine protease from seeds of Cucumis maderaspatensis, aimed with assessing the anticoagulant and antiplatelet activities. The purified serine protease was obtained by subjecting the seed extract to ammonium sulphate precipitation followed by anion exchange and gel filtration chromatography. Twenty seven-fold purification with the specific activity of 884.2 U/mg of protease activity was obtained. The characterization of the novel protease enzyme activity for optimum temperature, pH and effect of different protease inhibitors and metal ions were measured using caseinolytic assay and casein zymogram. The relative molecular mass of the novel neutral serine protease (CmSP) is ~ 32 kDa. Its anticoagulant was determined by assessing the delay in plasma re-calcification time in both platelet-rich and platelet-poor plasma. The antiplatelet activity of serine protease was demonstrated by inhibition of agonists induced platelet aggregation; it was in the order of Epinephrine > Adenosine tri phosphate. Further studies would decipher the mechanism of action to understand its therapeutic potential as an antiplatelet and anticoagulant molecule.

Facile-one pot-green synthesis, antibacterial, antifungal, antioxidant and antiplatelet activities of lignin capped silver nanoparticles: A promising therapeutic agent.

The current work portrays the green synthesis of Lignin Capped Silver Nanoparticles (LCSN) and their antibacterial, antifungal, antioxidant and antiplatelet potential. The LCSN was synthesized in water using a carbohydrate based polymer 'lignin' as the reducing and capping agents. The peak at 406nm (λmax) in the UV-Vis., spectrum and EDX analysis confirmed 1.68% (w/w) of silver was found to be loaded on lignin. The characteristic sharp peaks appeared in the PXRD spectrum showed fcc crystalline structure LCSN. SEM and TEM images indicated that the spherical Ag-NPs were well dispersed on lignin with an average particle size of ~10-15nm. LCSN showed antibacterial and antifungal activity against human pathogens S. aureus, E. coli and A. niger and the percentage of zone of inhibition was found to be 10%, 12% and 80% respectively. Further, LCSN was evaluated for antioxidant potential using DPPH scavenging assay, interestingly it showed antioxidant activity and the percentage against positive control vitamin C was found to be 70%. Furthermore, LCSN did not interfere in plasma coagulation; however, it found to inhibit agonist ADP induced platelet aggregation of human platelet rich plasma. The observed inhibition was found to be 37% and the calculated IC50 value was found to be 9mg/mL. LCSN did not lyses RBC membrane when assayed hemolytic activity suggesting its non-toxic nature.

Propensity of crocin to offset Vipera russelli venom induced oxidative stress mediated neutrophil apoptosis: a biochemical insight.

Viper envenomation results in inflammation at the bitten site as well as target organs. Neutrophils and other polymorphonuclear leukocytes execute inflammation resolving mechanism and will undergo apoptosis after completing the task. However, the target specific toxins induce neutrophil apoptosis at the bitten site and in circulation prior to their function, thus reducing their number. Circulating activated neutrophils are major source of inflammatory cytokines and leakage of reactive oxygen species (ROS)/other toxic intermediates resulting in aggravation of inflammatory response at the bitten/target site. Therefore, neutralization of venom induced neutrophil apoptosis reduces inflammation besides increasing the functional neutrophil population. Therefore, the present study investigates the venom induced perturbances in isolated human neutrophils and its neutralization by crocin (Crocus sativus) a potent antioxidant carotenoid. Human neutrophils on treatment with venom resulted in altered ROS generation, intracellular Ca(2+) mobilization, mitochondrial membrane depolarization, cyt-c translocation, caspase activation, phosphatidylserine externalization and DNA damage. On the other hand significant protection against oxidative stress and apoptosis were evidenced in crocin pre-treated groups. In conclusion the viper venom induces neutrophil apoptosis and results in aggravation of inflammation and tissue damage. The present study demands the necessity of an auxiliary therapy in addition to antivenin therapy to treat secondary/overlooked complications of envenomation.

Therapeutic drug-induced platelet apoptosis: an overlooked issue in pharmacotoxicology.

The surfacing of the applied fields of biology such as, biotechnology, pharmacology and drug discovery was a boon to the modern man. However, it had its share of disadvantages too. The indiscriminate use of antibiotics and other biological drugs resulted in numerous adverse reactions including thrombocytopenia. One of the reasons for drug-induced thrombocytopenia could be attributed to an enhanced rate of platelet apoptosis, which is a less investigated aspect. The present essay sheds light on the adverse (pro-apoptotic) effects of some of the commonly used drugs and antibiotics on platelets viz. cisplatin, aspirin, vancomycin and balhimycin. Furthermore, the undesirable reactions resulting from chemotherapy could be attributed at least to some extent to the systemic stress induced by microparticles, which in turn are the byproducts of platelet apoptosis. Thereby, the essay aims to highlight the challenges in the emerging trend of cross-disciplinary implications, i.e., drug-induced platelet apoptosis, which is a nascent field. Thus, the different mechanisms through which drugs induce platelet apoptosis are discussed, which also opens up a new perspective through which the adverse effects of commonly used drugs could be dealt. The drug-associated platelet toxicity is of grave concern and demands immediate attention. Besides, it would also be appealing to examine the platelet pro-apoptotic effects of other commonly used therapeutic drugs.

Alleviation of viper venom induced platelet apoptosis by crocin (Crocus sativus): implications for thrombocytopenia in viper bites.

Viper envenomations are characterized by prominent local and systemic manifestations including hematological alterations. Snake venom metalloproteinases (SVMPs) and phospholipase A2 (PLA2) plays crucial role in the pathophysiology of hemorrhage by targeting/altering the platelets function which may result in thrombocytopenia. Platelets undergo the classic events of mitochondria-mediated apoptotic pathway due to augmented endogenous reactive oxygen species (ROS) levels. The observed anticoagulant effects during viper envenomations could be due to exacerbated platelet apoptosis and thrombocytopenia. Moreover, antivenin treatments are ineffective against the venom-induced oxidative stress; therefore, it necessitates an auxiliary therapy involving antioxidants which can effectively scavenge the endothelium-generated/endogenous ROS and protect the platelets. The present study explored the effects of viper venom on platelet apoptosis and its amelioration by a phytochemical crocin. The study evaluated the Vipera russelli venom-induced apoptotic events including endogenous ROS generation, intracellular Ca(2+) mobilization, mitochondrial membrane depolarization, cyt-c translocation, caspase activation and phosphatidylserine externalization which were effectively mitigated when the venom was pre-treated with crocin. The study highlights one of the less studied features of venom-induced secondary complications i.e. platelet apoptosis and sheds light on the underlying basis for venom-induced thrombocytopenia, systemic hemorrhage and in vivo anticoagulant effect.

Vipera russelli venom-induced oxidative stress and hematological alterations: amelioration by crocin a dietary colorant.

Snakebite is a serious medical and socio-economic problem affecting the healthy individuals and agricultural and farming populations worldwide. In India, Vipera russelli snakebite is common, ensuing high morbidity and mortality. The venom components persuade multifactorial stress phenomenon and alter the physiological setting by causing disruption of the blood cells and vital organs. The present study demonstrates the anti-ophidian property of Crocin (Crocus sativus), a potent antioxidant against viper venom-induced oxidative stress. The in vivo oxidative damage induced by venom was clearly evidenced by the increased oxidative stress markers and antioxidant enzymes/molecules along with the proinflammatory cytokines including IL-1ß, TNF-α and IL-6. Furthermore, venom depleted the hemoglobin, hematocrit, mean corpuscular volume and platelet count in experimental animals. Crocin ameliorated the venom-induced oxidative stress, hematological alteration and proinflammatory cytokine levels. At present, administration of antivenom is an effective therapy against systemic toxicity, but it offers no protection against the rapidly spreading oxidative damage and infiltration of pro-inflammatory mediators. These pathologies will continue even after antivenom administration. Hence, a long-term auxiliary therapy is required to treat secondary as well as neglected complications of snakebite.

Differential action of phytochemicals on platelet apoptosis: a biological overview.

Platelets are anuclear blood cells originating from bone megakaryocytes. Despite being anuclear, their number is maintained by apoptosis, a process of programmed cell death. The rate of apoptotic death of platelets is accelerated by oxidative and shear stress, ex vivo storage (blood banking conditions) and certain pathophysiological disorders. These factors initiate apoptotic events through the mitochondria- mediated intrinsic pathway. Besides, apoptotic platelets also release phosphatidylserine-positive membrane fractions called microparticles, which cause fibrin deposition and thrombus formation, and are involved in the promulgation of a host of disease conditions including cardiovascular diseases. In this context, several phytochemicals have been reported to be cardioprotective and work by inhibiting platelet aggregation or by dissolving the fibrin clots. Besides, ample reports focus on the positive effects of phytochemicals on normal physiology of platelets, but do not focus on their adverse effects on platelets. Moreover, platelets are reported to be extremely sensitive to therapeutic components in the blood. For example, resveratrol and thymoquinone are hitherto known compounds to possess proapoptotic effects on platelets. In contrast, cinnamtannin B1 and crocin exhibit antiapoptotic effects. Thus, the current review aims to elucidate the underlying mechanisms through which the phytochemicals mediate their effects on platelet apoptosis. Moreover, the need for scrutiny of therapeutic compounds for their effects on platelet functions before including them in treatment regimen is also being emphasized.

Daboia russelli venom hyaluronidase: purification, characterization and inhibition by ß-3-(3-hydroxy-4-oxopyridyl) α-amino-propionic Acid.

The present study describes the purification and characterization of a hyaluronidase (DRHyal-II) from Daboia/Vipera russelli venom and its inhibition by ß-3-(3-hydroxy-4-oxopyridyl) α-amino-propionic acid, the mimosine. Gel permeation and ion exchange chromatography were employed to isolate DRHyal-II. The molecular mass by MALDITOF mass spectrometry was found to be 28.3 kDa. Single band in reduced SDS-PAGE suggested the monomeric nature. It was optimally active at pH 5.5 and at 37C and require 150 mM NaCl in the reaction mixture. It was specific to hyaluronan substrate and belongs to class-I or the neutral active enzymes. DRHyal-II was non-toxic by itself but, it potentiated the myotoxicity of VRV-PL-VIII myotoxin and hemorrhagic activity of hemorrhagic complex (HC). In in vitro experiments, mimosine inhibited the activity of DRHyal-II and the hyaluronidase activity of whole venom dose dependently. In in vivo experiments, mimosine inhibited the DRHyal-II potentiated myotoxicity of VRV-PL-VIII myotoxin and hemorrhagic activity of HC. The inhibition was due to the formation of DRHyal-II-mimosine inhibitory complex that resulted in significant structural changes at secondary and tertiary levels as evidenced by fluorescence emission and CD spectral studies. Hence, in this study an attempt was made to establish the possible role of hyaluronidase activity in the pathology of Daboia/Vipera russelli venom and the beneficial effects of its inhibition with special emphasis on the management of local toxicity.

The Hag-protease-II is a fibrin(ogen)ase from Hippasa agelenoides spider venom gland extract: purification, characterization and its role in hemostasis.

The current study describes the biochemical, biophysical and pharmacological properties of Hag-protease-II from Hippasa agelenoides spider venom gland extract. The Hag-protease-II was purified to homogeneity using gel filtration and ion-exchange chromatography. The molecular mass was found to be 28.749 kDa by MALDI-TOF mass spectrometry. PMSF abolished the activity while EDTA, EGTA, IAA and 1, 10-phenanthrolene did not. Hag-protease-II hydrolyzed casein, fibrinogen and fibrin, however it did not hydrolyze gelatin, fibronectin and collagen types- I and IV. It was non-lethal and devoid of hemorrhagic, myotoxic and edema forming activities. It dose dependently reduced re-calcification time of citrated human plasma. Strikingly; the Hag-protease-II coagulated the factor X deficient congenital human plasma. It hydrolyzed Bß-chain but, did not degrade Aα- and γ-chains of fibrinogen while, it hydrolyzed α-polymer and α-chain but not the ß-chain and γ-γ dimers of partially cross-linked fibrin clot. The Hag-protease-II induced aggregation of human platelets in PRP dose dependently, however it did not interfere in collagen induced aggregation of PRP and washed human platelets.

Factor Xa-like and fibrin(ogen)olytic activities of a serine protease from Hippasa agelenoides spider venom gland extract.

In the recent past, a low molecular mass serine protease, the Hag-protease that caused pro-coagulant activity and as well as local toxicity was isolated and characterized from the Hippasa agelenoides spider venom gland extract (Devaraja et al., Toxicon 52:130-138, 2008). In the current study, the pro-coagulant property has been investigated further and the results are presented. The Hag-protease reduced the re-calcification time of citrated human plasma. It reduced the activated partial thromboplastin time (APTT), and prothrombin time (PT) suggesting its participation in common pathway of blood coagulation. Interestingly, it coagulated the citrated human plasma in the absence of CaCl(2) but, it was lacking thrombin-like activity as it did not clot the purified fibrinogen. Strikingly, the enzyme coagulated the factor X deficient congenital human plasma, suggesting the factor Xa-like activity. However, the cumulative augmented activity was observed in presence of CaCl(2) and phospholipids. Further, the Hag-protease preferentially hydrolyzed the Aalpha chain and then the Bbeta-chain, but not the gamma-chain. As a result, truncated fibrinogen generated was lacking in the polymerization property. It hydrolyzed all the subunits of partially cross-liked fibrin clot (alpha-polymer, alpha-chain, beta-chain, and gamma-gamma dimers). Further, at low concentrations, the Hag-protease stimulated the aggregation of human platelets in platelet rich plasma, but at high concentrations caused spontaneous clumping. In contrast, it inhibited the collagen induced aggregation of washed human platelets. In summary, the present study for the first time reporting the factor Xa-like activity of a serine protease especially from the spider venom that exhibited opposing effects on hemostasis, the pro-coagulant activity and the anti-coagulant activity including fibrin(ogen)olytic and platelet aggregation inhibition activities.

Inhibition of local effects of Indian Daboia/Vipera russelli venom by the methanolic extract of grape (Vitis vinifera L.) seeds.

Although anti-venom therapy is available for the treatment of fatal bite by snakes, it offers less or no protection against the local effects such as dermo- and myonecrosis, edema, hemorrhage and inflammation at the bitten region. The viper species are known for their violent local effects and such effects have been commonly treated with plant extracts without any scientific validation in rural India. In this investigation, the methanolic extract of grapes (Vitis vinifera L.) seed was studied against the Indian Daboia/Vipera russelli venom-induced local effects. The extract abolished the proteolytic and hyaluronidase activities and also efficiently neutralized the hemorrhage, edema-inducing and myonecrotic properties of the venom. In addition, the extract also inhibited partially the pro-coagulant activity of the venom and abolished the degradation of Aalpha and Bbeta chains of human fibrinogen. Thus, the extract possesses potent anti-snake venom property, especially against the local effects of viper bites.

Effect of anticoagulants on the plasma hyaluronidase activities.

Mammalian hyaluronidases (HAases) are an endo-beta-N-acetyl-hexosaminidases that degrade hyaluronan (HA) and have been implicated in diverse pathophysiological functions. Several pathological conditions, such as diabetes, monoclonal gammapathy, and bladder and prostate tumors, report the distorted plasma HAase activity. However, the plasma HAase (hHyal-1) activity has been presumed to change with the circulating HA level and serves as an early marker for several diseases. It has been generally practised to use the anticoagulants such as tri-sodium citrate/di-sodium EDTA/heparin for the preparation of plasma for both biochemical and clinical analyses. In the present investigation, the effect of anticoagulants on plasma HAaseactivity was evaluated and compared with the serum HAase activity that is devoid of anticoagulants as no study provides information in this regard. The results suggested that the plasma HAase activity in the presence of the recommended concentration of EDTA was highly comparable/similar to that of the serum HAase activity. In contrast, citrated or heparinized plasma recorded a significantly reduced level of activity than that of the serum HAase activity. In conclusion, our results suggested that the EDTA-treated plasma samples are a better choice compared with heparin and citrated samples to assess the HAase activity.

A low molecular weight serine protease: Purification and characterization from Hippasa agelenoides (funnel web) spider venom gland extract.

Despite the long history [Kaiser, E., 1956. Enzymatic activity of spider venoms. In: Buckley, E.E., Porges, N. (Eds.), Venoms. American Association for the Advancement of Science, Washington, DC, pp. 91-93] on proteolytic activity, no study so far claims the isolation of a serine protease from the spider venom/venom gland extract. Therefore, the present study describes the isolation and characterization of a low molecular weight serine protease from Hippasa agelenoides venom gland extract. The protease (Hag-protease) was purified to homogeneity using the combination of gel-permeation and ion-exchange chromatography. The molecular mass was found to be 16.350 kDa by matrix-assisted laser desorption ionization time of flight (MALDI-TOF) mass spectrometry. Hag-protease was optimally active at pH 7.5 and temperature of 37 degrees C. PMSF abolished the enzyme activity while EDTA, EGTA, IAA, 1, 10-phenanthrolene did not. It hydrolyzed proteins such as casein, fibronectin and collagen type-I dose dependently but did not degrade gelatin and collagen type-IV. The isolated protease was non-lethal and devoid of hemorrhagic, myotoxic and edema forming activities. The light microscopy of Hag-protease treated skin tissue sections at the site of injection showed extensive damage of extracellular matrix (ECM) of hypodermis without causing any damage to blood vessels and capillaries. Similar damage of ECM of muscle tissue sections without affecting myocytes was noticed. Hag-protease was found to be procoagulant in property when studied plasma recalcification time.