IN VITRO ANTIPLATELET AND ANTICOAGULANT ACTIVITY OF INDIGENOUS VEGETABLES FROM SOUTHERN THAILAND

Objective: Epidemiological studies have indicated that diets rich in fruits and vegetables help reduce the risk of cardiovascular diseases (CVDs).However, data about the antithrombotic activity of local vegetables is rare. The objective of this study was to evaluate antiplatelet and anticoagulantactivity in indigenous vegetables with high phenolic compounds collected from Southern Thailand.Methods: Five selected indigenous vegetables were crudely extracted by distilled water and 80% methanol. The extracts were screened for in vitroantiplatelet and anticoagulant activity at a concentration of 10 μg/μl. The antiplatelet activity was measured by inhibition of platelet adhesion tocollagen and thrombin-induced platelet aggregation, while the anticoagulant activity was assessed by the prothrombin time (PT) and activated partialthromboplastin time (APTT) tests.Results: Among the selected vegetables, the extracts of mon-pu (Glochidion perakense Hook.f.) and young cashew leaves (Anacardium occidentale L.)showed high antithrombotic properties. The highest antithrombotic activity was observed in the methanolic extract of mon-pu, which showed92.79±0.78% of platelet adhesion inhibition, 102.9±1.53% of platelet aggregation inhibition, and a prolonged APTT assay (48.92±0.94 s). Theprolonged APTT but normal PT results suggested that the extract could affect factors VIII, IX, XI, and XII of the intrinsic coagulation pathway.Conclusion: Our findings demonstrated antiplatelet and anticoagulation properties of indigenous vegetables from Southern Thailand. The multipotentialeffects of mon-pu extracts on antithrombosis evidently suggest that mon-pu can be considered as an excellent nutraceutical option in theprevention of thrombosis-related CVDs caused by different mechanisms

A survey of jellyfish sting knowledge among Thai divers in Thailand

Background: In tropical regions, jellyfish envenomation is a persistent hazard for people who spend time in the sea. Jellyfish stings can be dangerous, and among the people who face the greatest risk are scuba divers. This study therefore sought to determine the level of knowledge divers in Thailand have about the threat of jellyfish envenomation.  Materials and methods: In April 2018, a total of 238 divers responded to a questionnaire, thereby providing data for further statistical analysis. Results: The findings revealed that 31.91% of the study participants cited jellyfish stings as their most frequently encountered injury, with 68.09% having personal experience of the problem, or having seen others injured by jellyfish. However, 34.03% of the sample respondents believed their own level of knowledge to be “low” or “none”. The mean score was 71%, which can be considered satisfactory, but the scores for items concerning the recognition of signs of envenomation and items about first aid responses (52.74% and 59.13%, respectively) were not acceptable.  Conclusions: Divers frequently experience jellyfish stings, and diving personnel were highly rated for their knowledge in this area. However, very few were fully confident in their first aid capabilities, and therefore it can be argued that it is necessary to improve the level of medical education and to provide training to eliminate this weakness.

IN VITRO ANTICOAGULANT AND ANTIOXIDANT ACTIVITIES OF PRASAPLAI RECIPE AND ZINGIBER CASSUMUNAR ROXB. EXTRACTS

Objective: This present study aimed to evaluate the anticoagulant activity and antioxidant properties of Prasaplai recipe (PPR), a Thai traditionalmedicine, and its major ingredient, Zingiber cassumunar (ZC) Roxb. extracts, seeking new therapeutic purposes for the recipe.Methods: Aqueous extracts of PPR and ZC Roxb. were prepared by hot water decoction technique. The anticoagulant activity of the extracts wasevaluated by prothrombin time (PT) and activated partial thromboplastin time (APTT) tests. In addition to anticoagulant activity, total phenolcontent and antioxidant activity were investigated. Total phenol content was determined using the Folin–Ciocalteu assay. The antioxidant activity wasestimated by DPPH radical scavenging activity and ferric reducing antioxidant power assay.Results: The APTT of plasma samples mixed with the PPR and ZC Roxb. extracts was significantly prolonged (p<0.05) at the concentration of1.0 mg/ml and above comparing to the control (normal saline solution) but was no significantly different for the PT. These results suggested thatPPR and ZC Roxb. extracts showed anticoagulant activity affecting the function of coagulation factor in the intrinsic pathway. All aqueous extractspossessed considerable antioxidant activity and were rich in total polyphenol.Conclusion: This finding indicates that the aqueous extracts possess significant anticoagulant and antioxidant activities, thus showing the potentialPPR and ZC Roxb. as a new source of bioactive compounds for therapeutic purposes, with particular emphasis on the prevention and treatment ofthrombosis

Extracellular Vesicle-Mediated Macrophage Activation: An Insight Into the Mechanism of Thioredoxin-Mediated Immune Activation

Extracellular vesicles (EVs) generated from redox active anticancer drugs are released into the extracellular environment. These EVs contain oxidized molecules and trigger inflammatory responses by macrophages. Using a mouse model of doxorubicin (DOX)-induced tissue injury, we previously found that the major sources of circulating EVs are from heart and liver, organs that are differentially affected by DOX. Here, we investigated the effects of EVs from cardiomyocytes and those from hepatocytes on macrophage activation. EVs from H9c2 rat cardiomyocytes (H9c2 EVs) and EVs from FL83b mouse hepatocytes (FL83 b EVs) have different levels of protein-bound 4-hydroxynonenal and thus different immunostimulatory effects on mouse RAW264.7 macrophages. H9c2 EVs but not FL83 b EVs induced both pro-inflammatory and anti-inflammatory macrophage activation, mediated by NFκB and Nrf-2 pathways, respectively. DOX enhanced the effects of H9c2 EVs but not FL83 b EVs. While EVs from DOX-treated H9c2 cells (H9c2 DOXEVs) suppressed mitochondrial respiration and increased glycolysis of macrophages, EVs from DOX-treated FL83b cells (FL83b DOXEVs) enhanced mitochondrial reserve capacity. Mechanistically, the different immunostimulatory functions of H9c2 EVs and FL83 b EVs are regulated, in part, by the redox status of the cytoplasmic thioredoxin 1 (Trx1) of macrophages. H9c2 DOXEVs lowered the level of reduced Trx1 in cytoplasm while FL83b DOXEVs did the opposite. Trx1 overexpression alleviated the effect of H9c2 DOXEVs on NFκB and Nrf-2 activation and prevented the upregulation of their target genes. Our findings identify EVs as a novel Trx1-mediated redox mediator of immune response, which greatly enhances our understanding of innate immune responses during cancer therapy

Extracellular Vesicles Released After Cranial Radiation: An Insight into an Early Mechanism of Brain Injury

Cranial radiation is important for treating both primary brain tumors and brain metastases. A potential delayed side effect of cranial radiation is neurocognitive function decline. Early detection of CNS injury might prevent further neuronal damage. Extracellular vesicles (EVs) have emerged as a potential diagnostic tool because of their unique membranous characteristics and cargos. We investigated whether EVs can be an early indicator of CNS injury by giving C57BJ/6 mice 10 Gy cranial IR. EVs were isolated from sera to quantify: 1) number of EVs using nanoparticle tracking analysis (NTA); 2) Glial fibrillary acidic protein (GFAP), an astrocyte marker; and 3) protein-bound 4-hydroxy-2-nonenal (HNE) adducts, an oxidative damage marker. Brain tissues were prepared for immunohistochemistry staining and protein immunoblotting. The results demonstrate: 1) increased GFAP levels (p \u3c 0.05) in EVs, but not brain tissue, in the IR group; and 2) increased HNE-bound protein adduction levels (p \u3c 0.05). The results support using EVs as an early indicator of cancer therapy-induced neuronal injury

ANTI-COAGULANT PROPERTIES OF FLAVONOID COMPOUNDS: POTENTIAL STRUCTURE-FUNCTIONAL RELATIONSHIP

Objective: Flavonoids, naturally-occurring compounds in fruits and vegetables, possess anti-coagulant property. However, a very few studies wereattempted to understand how flavonoid structure influences its anti-coagulation property, such as clotting time. In this study, we investigatedstructurally similar flavonoid compounds which differ in the number of hydroxyl groups and compared their anti-coagulation properties.Methods: We selected and evaluated five flavonoid compounds, that is, chrysin, apigenin, luteolin, kaempferol, and quercetin, for their anti-coagulantproperties using in vitro prothrombin time (PT) assays and activated partial thromboplastin time (APTT) assay.Results: Our findings suggested that quercetin, kaempferol, and luteolin showed a significant anti-coagulant effect on APTT (p<0.05) in a dosedependentmanner. The dose of 500 μM quercetin showed potent prolong APTT with 37.43±1.60 s, followed by 500 μM of kaempferol and luteolin(34.63±1.29 s and 4.83±1.56 s, respectively). Furthermore, a combination of 500 μM of quercetin with 0.25 U/ml of heparin demonstrated prolongAPTT (52.16±5.18 s) when compared with individual effects of either 0.25 U/ml heparin (33.4±0.50 s) or 500 μM quercetin (37.43±1.62 s) alone.Conclusion: Our results demonstrated that numbers of the hydroxyl group on flavonoid compounds influence anti-coagulation properties. Inaddition, the prolonged APTT assay results suggested that quercetin, kaempferol, and luteolin could affect factors VIII, IX, XI, and XII of intrinsicpathway. Moreover, the synergistic effect of quercetin further enhances the heparin anti-coagulation effect. Based on our findings, we recommendthat the consumption of vegetables and fruits rich in quercetin, luteolin, and kaempferol could help prevent thrombotic stroke in high-risk patients

A Systematic Review and Meta-Analysis of the Proportion Estimates of Disseminated Intravascular Coagulation (DIC) in Malaria

Disseminated intravascular coagulation (DIC) is a potentially life-threatening condition that causes systemic coagulation to be turned on and coagulation factors to be used up. However, the evidence for DIC in malaria patients is still not clear, and small case series and retrospective studies have shown varying results. This meta-analysis was intended for the evaluation of the evidence of DIC among malaria patients using a meta-analysis approach. The protocol for the systematic review was registered at PROSPERO as CRD42023392194. Studies that investigated DIC in patients with malaria were searched in Ovid, Scopus, Embase, PubMed, and MEDLINE. The pooled proportion with 95% confidence intervals (CI) of DIC among malaria patients was estimated using a random-effects model. A total of 1837 articles were identified, and 38 articles were included in the meta-analysis. The overall proportion of DIC in malaria was 11.6% (95% CI: 8.9%–14.3%, I2: 93.2%, 38 studies). DIC in severe falciparum malaria and fatal malaria was 14.6% (95% CI: 5.0–24.3%, I2: 95.5%, 11 studies) and 82.2% (95% CI: 56.2–100%, I2: 87.3, 4 studies). The estimates of DIC among severe malaria patients who had multi-organ dysfunction with bleeding, cerebral malaria, acute renal failure, and ≥2 complications were 79.6% (95% CI: 67.1–88.2%, one study), 11.9% (95% CI: 7.9–17.6%, one study), 16.7% (95% CI: 10.2–23.3%, ten studies), and 4.8% (95% CI: 1.9–7.7%, nine studies), respectively. The proportion estimates of DIC among the patients with malaria depended on the Plasmodium species, clinical severity, and types of severe complications. The information from this study provided useful information to guide the management of malaria patients. Future studies are needed to investigate the association between Plasmodium infection and DIC and to understand the mechanism of malaria-induced DIC

Evaluation of Thrombolytic Activity of Zingiber cassumunar Roxb. and Thai Herbal Prasaplai Formula

The propose of this study was to investigate in vitro thrombolytic activity of Zingiber cassumunar Roxb. and Prasaplai, a Thai herbal formulation of Z. cassumunar Roxb. Herbs were extracted with boiling water and concentrated by lyophilization. To observe their thrombolytic potential, an in vitro clot lysis method was applied where streptokinase and sterile distilled water were used as positive and negative controls, respectively. Crude aqueous extracts from Z. cassumunar Roxb. and Prasaplai formula showed significant thrombolytic activity by clot lysis of 17.90% and 25.21%, respectively, compared to the negative control water (5.16%) while the standard streptokinase revealed 64.78% clot lysis. These findings suggest that Z. cassumunar Roxb. exhibits moderate thrombolytic activity and cloud play an important role in the thrombolytic properties of Prasaplai formula. However, further study should be done to observe in vivo clot dissolving potential and to isolate active component(s) of these extracts

Clinical severity of β-thalassaemia/Hb E disease is associated with differential activities of the calpain-calpastatin proteolytic system.

Earlier observations in the literature suggest that proteolytic degradation of excess unmatched α-globin chains reduces their accumulation and precipitation in β-thalassaemia erythroid precursor cells and have linked this proteolytic degradation to the activity of calpain protease. The aim of this study was to correlate the activity of calpain and its inhibitor, calpastatin, with different degrees of disease severity in β-thalassaemia. CD34(+) cells were enriched from peripheral blood of healthy individuals (control group) and patients with mild and severe clinical presentations of β(0)-thalassaemia/Hb E disease. By ex vivo cultivation promoting erythroid cell differentiation for 7 days, proerythroblasts, were employed for the functional characterization of the calpain-calpastatin proteolytic system. In comparison to the control group, enzymatic activity and protein amounts of μ-calpain were found to be more than 3-fold increased in proerythroblasts from patients with mild clinical symptoms, whereas no significant difference was observed in patients with severe clinical symptoms. Furthermore, a 1.6-fold decrease of calpastatin activity and 3.2-fold accumulation of a 34 kDa calpain-mediated degradation product of calpastatin were observed in patients with mild clinical symptoms. The increased activity of calpain may be involved in the removal of excess α-globin chains contributing to a lower degree of disease severity in patients with mild clinical symptoms

Identification of active calpastatin molecular species.

<p><b>A</b>) Activity profiles of samples from 10⁶ erythroid precursor cells isolated from control and patients with mild and severe clinical symptoms, respectively. Numbers at the bottom refer to the molecular weight of proteins present in the gel slices. <b>B</b>) Bar graph presentation of calpastatin activity in extracted fractions of SDS-PAGE gels. Activity is presented as percentage of total calpastatin activity measured in the sample. White, grey, and black bars represent activity of calpastatin forms in samples from control, mild, and severe clinical symptoms, respectively. Asterisks above the bars indicate a significance of <i>p</i><0.05.</p