Platelet Aggregation Activity Research Articles

Genome-based taxonomic identification and safety assessment of an Enterococcus strain isolated from a homemade dairy product.

The aim of this study was to explore the taxonomic identification and evaluate the safety of a bacterium, Enterococcus lactis IDCC 2105, isolated from homemade cheese in Korea, using whole genome sequence (WGS) analysis. It sought to identify the species level of this Enterococcus spp., assess its antibiotic resistance, and evaluate its virulence potential. WGS analysis confirmed the bacterial strain IDCC 2105 as E. lactis and identified genes responsible for resistance to erythromycin and clindamycin, specifically msrC, and eatAv, which are chromosomally located, indicating a minimal risk for horizontal gene transfer. The absence of plasmids in E. lactis IDCC 2105 further diminishes the likelihood of resistance gene dissemination. Additionally, our investigation into seven virulence factors, including hemolysis, platelet aggregation, biofilm formation, hyaluronidase, gelatinase, ammonia production, and β-glucuronidase activity, revealed no detectable virulence traits. Although bioinformatic analysis suggested the presence of collagen adhesion genes acm and scm, these were not corroborated by phenotypic virulence assays. Based on these findings, E. lactis IDCC 2105 presents as a safe strain for potential applications, contributing valuable information on its taxonomy, antibiotic resistance profile, and lack of virulence factors, supporting its use in food products.

Competitive Analysis of the Binding Affinity of Montelukast, Zafirlukast and Gemilukast to CysLTR1, P2Y12 and PPAR-γ and their Possible Cardioprotective Effect: Using in silico Methods

Background Asthma is a very common respiratory disorder, affecting more than 360 million people worldwide. It is a chronic inflammatory disorder of the airways with the symptoms of shortness of breath, coughing, chest tightness, wheezing, and sometimes chest pain. Leukotrienes play an important role in bronchoconstriction during the allergen or exercise-induced acute asthma attack. Aim The study aims to predict the interactions between leukotriene antagonist drugs and CysLT receptor-1 (CysLTR1), P2Y12 and peroxisome proliferator-activated receptor gamma (PPAR-γ) on a competitive basis. The study also has the objective of understanding the cardioprotective roles of the drugs. Introduction Asthma is strongly linked to the development of acute coronary syndrome by the leukotriene-induced activation of CysLTR1, platelet aggregation and thrombosis by activating P2Y12. PPAR-γ is considered to show benefits against atherosclerosis, diabetes, hypertension, obesity and dyslipidaemia, which are risk factors for the development of cardiovascular disorders. Leukotriene receptor inhibitors act with these three types of receptors to show therapeutic effects. Materials and Methods To predict the possible interactions between the drugs and the receptors, the study has used in silico methods. Results and Discussion Montelukast, Zafirlukast and Gemilukast are potential antagonists of CysLTR1 and P2Y12. They are also responsible for the upregulation of PPAR-γ. Thus, these drugs show a cardioprotective role in asthma-induced cardiac disorders. Conclusion A competitive in silico study of Montelukast, Zafirlukast and Gemilukast to predict their binding to CysLTR1, P2Y12 & PPAR-γ revealed that Montelukast is more effective than the other two drugs for showing a cardioprotective role.

Proinflammatory biomarkers and platelet aggregation activity in patients with coronary artery disease

Proinflammatory biomarkers and platelet aggregation activity in patients with coronary artery disease

Spontaneous and stimulated platelet aggregation activity in patients with atrial fibrillation and thrombotic complications

Aim: To study spontaneous and stimulated by adenosine diphosphate (ADP), adrenaline and collagen platelet aggregation activity, CD40-CD40 ligand system in patients with atrial fibrillation (AF) with previous thromboembolic complications (TEC) and newly diagnosed thrombosis.Material and Methods. The study included patients with «non-valvular» AF and anamnesis of TEC and newly identified thrombosis on anticoagulants in the period 01.2020-01.2023: 13 patients had anamnesis of TEC (group 1), in 18 patients were initially diagnosed thrombosis/spontaneous atrial echocontrast of high grade (group 2). The comparison group was the healthy individuals (n = 31) (group 3). The spontaneous aggregation ability of platelets and stimulated with solutions of ADP, adrenaline and collagen, the level of CD40 markers, soluble CD40 ligand (CD40L) were studied.Results. The spontaneous platelet aggregation activity was significantly higher in patients of group 2 compared with healthy individuals with a formation trend of larger aggregates in patients with TEC. When aggregation was induced by collagen, the rate of aggregate formation was significantly lower in the 1st and 2nd groups compared with healthy individuals. When using ADP, a comparable decrease in the degree and speed was noted, and when using adrenaline, only the rate of platelet aggregation was observed in groups 1 and 2. The concentration of soluble CD40L was significantly higher in patients with a history of TEC and newly thrombosis.Conclusion. Spontaneous platelet aggregation activity and the concentration of soluble CD40L are increased both in patients with past of TEC and in patients with newly diagnosed thrombosis due to AF and anticoagulant monotherapy.

Role of <i>ITGB3</i>, <i>GP1B1</i>, and <i>ITGA2</i> gene polymorphisms in platelet dysfunction in patients with COVID-19-associated lung damage

The aim of the work. To investigate platelet aggregation, polymorphism in the genes that ensure its implementation, and the association between these indicators in patients with COVID-19-associated lung damage, depending on the severity of the clinical course. Methodology. The study involved 75 patients with COVID-19, which, depending on the severity of lung involvement, were divided into two groups: patients with damage of up to 50 % of the lung parenchyma (n = 48) and with damage of more than 50 % (n = 27) respectively. The control group consisted of healthy people (n = 24), comparable in sex and age. In all individuals, the number of platelets, platelet aggregation induced by ADP, collagen and ristomycin were studied; polymorphisms rs6065 in the GP1BA gene, rs1126643 in the ITGA2 gene, and rs5918 in the ITGB3 gene were determined by polymerase chain reaction. The analysis of the obtained data was executed using the IBM SPSS Statistics v. 23 (IMB Corp., USA). Results and discussion. In patients with moderate and severe COVID-19-associated lung damage, platelet aggregation induced by ADP, collagen, and ristomycin accelerated; in severe cases, the number of platelets decreased. The frequency of variants of the rs6065 polymorphism did not change, the frequency of occurrence of the T/C genotype of the rs5918 polymorphism increased; with moderate severity, the frequency of occurrence of the C/T and T/T genotypes of the rs1126643 polymorphism increased; with severe lung damage, the frequency of occurrence of the mutant C/C genotype polymorphism rs5918 increased. In moderate lung damage, the presence of the mutant T/T polymorphism rs1126643 accelerated collagen-induced platelet aggregation; in severe cases, the presence of mutant C/C and heterozygous variant C/T polymorphism rs5918 accelerated ADP-induced platelet aggregation. There was no effect of the rs6065 polymorphism on platelet aggregation. The data obtained indicate the possible role of genetic predisposition in the activation of platelet aggregation in patients with COVID-19-associated lung damage.

L-carnitine contributes to enhancement of viability and quality of platelet concentrates through changing the apoptotic and anti-apoptotic associated microRNAs

BackgroundMicro RNAs are known as the main regulator of messenger RNA translation in platelets and have a vital role in process of apoptosis during platelet storage. Our pervious study revealed that the expression of miR-145 and miR-326 changed significantly in platelets under maintenance conditions. This study aimed to evaluate the effect of L-carnitine (LC) as an additive to augment platelet quality by changing the microRNA expression. MethodsWe used ten platelet concentrate (PC) bags and divided each into two equal parts, LC- treated, and LC free PC. The expression of miR-145 and miR-326 were determined using real-time PCR. Moreover, we measured platelet count, platelet aggregation, platelet viability, and lactate dehydrogenase activity in all samples. ResultsThe miR-326 expression significantly increased during platelet storage with mean fold changes of 3.2 for the control and 2.5 for LC- treated PC. The mean fold changes in miR-145 expression was less in the control PC (0.52) compared to the LC- treated PC (0.79). Increased levels of platelet count, platelet aggregation, and platelet viability were found in the LC-treated compared to the untreated PC. ConclusionLC has a protective effect on platelet apoptosis, reduces the expression of apoptotic microRNA, and prevents the reduction of anti-apoptotic microRNA.

A novel GATA1 variant p.G229D causing the defect of procoagulant platelet formation

IntroductionGATA1 is one of the master transcription factors in hematopoietic lineages development which is crucial for megakaryocytic differentiation and maturation. Previous studies have shown that distinct GATA1 variants are associated with varying severities of macrothrombocytopenia and platelet dysfunction. ObjectiveTo determine the underlying pathological mechanisms of a novel GATA1 variant (c. 686G > A, p. G229D) in a patient with recurrent traumatic muscle hematomas. MethodsComprehensive phenotypic analysis of the patient platelets was performed. Procoagulant platelet formation and function were detected using flow cytometry assay and thrombin generation test (TGT), respectively. The ANO6 expression was measured by qPCR and western blot. The intracellular supramaximal calcium flux was detected by Fluo-5N fluorescent assay. ResultsThe patient displayed mild macrothrombocytopenia with defects of platelet granules, aggregation, and integrin αIIbβ3 activation. The percentage of the procoagulant platelet formation of the patient upon the stimulation of thrombin plus collagen was lower than that of the healthy controls (40.9 % vs 49.0 % ± 5.1 %). The patient platelets exhibited a marked reduction of thrombin generation in platelet rich plasma TGT compared to the healthy controls (peak value: ∼70 % of the healthy controls; the endogenous thrombin potential: ∼40 % of the healthy controls). The expression of ANO6 and intracellular calcium flux were impaired, which together with abnormal granules of the patient platelets might contribute to defect of procoagulant platelet function. ConclusionsThe G229D variant could lead to a novel platelet phenotype characterized by defective procoagulant platelet formation and function, which extended the range of GATA1 variants associated platelet disorders.

Safety of Tailored Transfemoral Carotid Artery Stenting for Symptomatic Elderly Patients: A Single Center Observational Study

Transfemoral carotid artery stenting (TFCAS) in symptomatic elderly patients (≥70 years old) may have a high periprocedural stroke rate. This study was performed to examine whether tailored TFCAS for symptomatic elderly patients is as safe as that for symptomatic nonelderly patients. The subjects were 185 patients with symptomatic internal carotid artery stenosis. Tailored TFCAS including postoperative management was performed based on preoperative examinations of vascular anatomy, plaque imaging, platelet aggregation activity, and cerebral hemodynamic impairment. The major 30-day perioperative stroke rates were examined. The patients included 51 (27.6%) <70 (group Y) and 134 (72.4%) ≥70 (group E) years old. Group E included significantly more cases with an elongated aortic arch, tortuous target lesion, and longer plaques (all P < 0.05). Among all cases, 181 (97.8%) procedures were performed as per preoperative planning. Group E had more frequent use of a proximal embolic protection device and a closed-cell or dual-layer micromesh stent (all P < 0.05). Seven patients (3.8%) had major stroke. Rates of major ischemic stroke (2.0% vs. 3.0%, P= 1.00) and intracranial hemorrhage (2.0% vs. 0.8%, P= 0.48) were low and did not differ significantly between groups Y and E. Symptomatic elderly patients have several unfavorable factors. However, tailored TFCAS for each patient based on preoperative examinations in symptomatic elderly patients may be as safe as that in symptomatic nonelderly patients.

Exosome Liberation by Human Neutrophils under L-Amino Acid Oxidase of Calloselasma rhodostoma Venom Action.

L-Amino acid oxidase (LAAO) is an enzyme found in snake venom that has multifaceted effects, including the generation of hydrogen peroxide (H2O2) during oxidative reactions, leading to various biological and pharmacological outcomes such as apoptosis, cytotoxicity, modulation of platelet aggregation, hemorrhage, and neutrophil activation. Human neutrophils respond to LAAO by enhancing chemotaxis, and phagocytosis, and releasing reactive oxygen species (ROS) and pro-inflammatory mediators. Exosomes cellular nanovesicles play vital roles in intercellular communication, including immune responses. This study investigates the impact of Calloselasma rhodostoma snake venom-derived LAAO (Cr-LAAO) on human neutrophil exosome release, including activation patterns, exosome formation, and content. Neutrophils isolated from healthy donors were stimulated with Cr-LAAO (100 μg/mL) for 3 h, followed by exosome isolation and analysis. Results show that Cr-LAAO induces the release of exosomes with distinct protein content compared to the negative control. Proteomic analysis reveals proteins related to the regulation of immune responses and blood coagulation. This study uncovers Cr-LAAO's ability to activate human neutrophils, leading to exosome release and facilitating intercellular communication, offering insights into potential therapeutic approaches for inflammatory and immunological disorders.

The effect of γ-irradiation at a dose of 1 Gy on the adenylate- and guanylate cyclase systems of rat platelets at different times of the post-radiation period

Objective. To study the effect of total γ-irradiation of the organism of rats at a dose of 1 Gy on the adenylate- and guanylate cyclase systems and their relationship with Ca2+ levels in the platelet cytoplasm.Materials and methods. The studies were carried out on outbred white male rats of mature age (6-7 months) weighing 250±30 g. Animals were irradiated (once and evenly) on the IGUR unit with 137Сs γ-quanta at a dose of 1 Gy (dose rate 0.62 Gy/min, for 1.61 min). Animals of the corresponding age served as controls. To determine the content of cyclic nucleotides, RIO cAMP/cGMP-iodine125-M-IBOX reagent kits were used. The amount of calcium in platelets was determined using a Fura-2/AM fluorescent probe using spectrofluorimeter SM 2203 “SOLAR” (Minsk, Belarus).Results. In the short period after irradiation, an increase in the activity of the adenylate cyclase system is observed. The cAMP level rises 1.8-1.5 times during 3-30 days of the post-radiation period. However, the increase in [Ca2+]cyt on the 3rd day after irradiation leads to its predominance by 1.8 times in relation to cAMP, which explains the appearance of an increased aggregation activity of the blood platelets in the immediate post-radiation period. An increase in the activity of the guanylate cyclase system and an increase in the intracellular content of cGMP are observed on the 90th day, which leads to a decrease in the Ca2+/cGMP ratio by 1.83 times and corresponds to a reduced platelet aggregation activity and the probability of bleeding in the long term of the rehabilitation period.Conclusion. The effects of radiation on the platelets of rats irradiated at a dose of 1 Gy are due to systemic disorders of the main intracellular regulatory mechanisms: Ca2+, cAMP, cGMP. Based on the data obtained, it can be concluded that pharmacological correction in the short-term after irradiation should be aimed at inhibiting the effects of [Ca2+]cyt and adenylate cyclase system activity, and in the long-term, at inhibition of guanylate cyclase system activity.

Anti-thrombotic Effects Mediated by a Novel Dual-Target Peptide Inhibiting Both Platelet Aggregation and Thrombin Activity without Causing Bleeding.

Classical anticoagulants and antiplatelets are associated with high frequencies of bleeding complications or treatment failure when used as single agents. Thrombin plays an important role in the blood coagulation system. GP IIb/IIIa is the central receptor of platelets, which can recognize the Arg-Gly-Asp (RGD) sequence and activate platelets. Molecular simulation and homology modeling were performed to design a novel dual-target anticoagulant short peptide (PTIP ). The activities of PTIP on coagulation and platelet in vitro were analyzed. The antithrombotic activity of PTIP was determined by pulmonary thromboembolism model, ferric chloride injury model and arteriovenous bypass thrombosis model. Bleeding effect and toxicity of PTIP were evaluated. We have constructed a novel dual-target peptide (PTIP) based on the direct thrombin inhibitor peptide (DTIP). PTIP was expressed at high levels in Pichia pastoris. PTIP interfered with thrombin-mediated coagulation and ADP-induced platelet aggregation in vitro. When injected intravenously or subcutaneously, PTIP showed potent and dose-dependent extension of aPTT and PT which were similar to DTIP; but only PTIP was capable of inhibiting platelet aggregation. PTIP (1.0 mg/kg) decelerated thrombosis formation in venous and arterial vessels induced by FeCl3 injury. PTIP (1.0 mg/kg) also prevented deep venous thrombosis and increased the survival rate associated with pulmonary thromboembolism. And PTIP effectively reduced thrombus length in arteriovenous bypass thrombosis model. Moreover, the antithrombotic dose of PTIP could not induce bleeding. These data establish that PTIP represents a novel antithrombotic agent whose effects involve both inhibition of platelet activation and reduction of fibrin generation. And PTIP not only can be used in venous thrombosis and arterial thrombosis, it can also replace the combined treatment of antiplatelet and anticoagulant drugs in thrombotic diseases.

Resveratrol in Neurodegeneration, in Neurodegenerative Diseases, and in the Redox Biology of the Mitochondria.

Neurodegeneration is a process leading to the progressive loss of structure and functions of neurons. Many neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, and Huntington's disease have shown many common points at the subcellular level. Neurons are metabolically active cells and need a high amount of energy. Mitochondria are known as the energy synthesis center for cells, involved in the synthesis of adenosine triphosphate by oxidative phosphorylation. Rather than just being an energy synthesis center, it has critical importance for many cellular functions such as calcium homeostasis, cell proliferation, cell growth, and apoptosis. In the process of mitochondrial dysfunction, cellular functions are disrupted and cells enter the apoptotic or necrotic pathway. Resveratrol (trans-3,5,4-trihydoxystilbene), a plant-derived polyphenol found in the seed of grapes, berries, peanuts, and wine, has many biological effects such as inhibition of lipid peroxidation, scavenging of free radicals, changes in eicosanoid synthesis, inhibition of platelet aggregation, anti-inflammatory and anticancer activity, and regulation of lipid metabolism. Through the reviewed literature, the current study investigated the protective role of resveratrol in neurodegenerative diseases. Studies show that resveratrol moderates mitochondrial function, redox status, and cellular dynamics in both in vivo and in vitro experimental models of neurodegeneration. Resveratrol suppresses reactive oxygen species production by reducing the activity of complex III due to its competition effect with coenzyme Q. In the present work, we discussed the protective effects of resveratrol on neurodegeneration, neurodegenerative diseases, and the redox biology of the mitochondria.

#4878 STUDYING THE EFFECTIVENESS OF PRASUGREL AND CLOPIDOGEL IN PATIENTS WITH CHRONIC LOWER LIMB ISCHEMIA WITH STAGE 2 CKD

Abstract Background and Aims To compare the clinical and rheological efficacy of prasugrel and clopidogrel in patients with chronic lower limb ischemia with stage 2 chronic kidney disease. According to statistics, in Europe, peripheral arterial disease is common in more than 40 million Europeans. According to the developed tactics, not only stroke should be recognized as a serious, life-threatening condition, but other peripheral vascular diseases can lead to disability and death. Methods The study included 101 patients with critical lower limb ischemia of atherosclerotic origin. The mean age of the patients was 51.51±8.33 years. As a control group, 20 healthy volunteers (mean age 53.90 ± 9.05 years) were examined without signs of pathology of the cardiovascular system, including those of a vegetative nature. All patients included in the study were hospitalized in the Department of Interventional Cardiology of the Republican Specialized Scientific and Practical Center for Therapy and Medical Rehabilitation, Tashkent city. Results In 94 patients (93.07%), CLCI was accompanied by hemodynamically significant multifocal atherosclerotic lesions. The first place in frequency of combined lesions was occupied by the coronary vessels, the second place - by the arteries of the brachiocephalic basin. 82 patients (81.19%) had previously suffered arterial occlusion, also most often in the form of acute myocardial infarction, somewhat less often - acute cerebrovascular accident. 47 patients (46.53%) underwent revascularization earlier, mainly coronary artery stenting, coronary artery bypass grafting, carotid endarterectomy. 8 patients (7.92%) underwent revascularization of the contralateral leg. The study studied spontaneous (SAT) and ADP-induced platelet aggregation. In the CG, SAT was noted only in 50% of cases and in all patients of the CLCI group. After comparing the results in the groups of patients with CLCI and representatives of the CG, in whom SAT was noted, it was found that both the rate and degree of SAT in the group of patients were significantly higher than in healthy individuals (2.95±0.59% versus 0.55± 0.21% and 6.05±1.54% versus 1.24±0.21%, respectively, p&amp;lt;0.001 significance of the intergroup difference of both parameters). All patients due to CINC underwent endovascular revascularization of the affected leg. After the procedure, patients were prescribed an antiplatelet agent at a maintenance dose (prasugrel 10 mg/day, clopidogrel 75 mg/day). After 3 months, a control examination was carried out, during which the dynamics of platelet aggregation activity was studied. It was found that the indicators of CAT and ADP induced aggregation decreased by 22-37%. During the test, patients with resistance to antiplatelet agents were identified - in whom the decrease in the degree of ADP5 induced aggregation was less than 20%. Such resistance was found in 19 patients: 1 patient in the prasugrel group (1.69%) and 18 patients in the clopidogrel group (42.86%, chi square = 27.00, p&amp;lt;0.001). Isolation of patients with normal and reduced response to the loading dose of clopidogrel revealed some differences (Table 3.3). So, already initially in this group of patients, the degree of ADP5-induced aggregation was lower than in the group of patients with an antiplatelet response of more than 20%. After the application of a loading dose of clopidogrel, differences in relative dynamics were noted only in terms of the degree of ADP%-induced aggregation (p&amp;lt;0.001). Conclusions Thus, the present study revealed that in patients with CLCI, there is a significant activation of CAT and an increase in the aggregation response to the introduction of high doses of the inducer of ADP5 aggregation. The loading dose of antiplatelet agents contributed to a significant decrease in platelet aggregation activity after 2 hours with a large effect of prasugrel. Chronic antiplatelet therapy led to a decrease in the rate and degree of aggregation, both spontaneous and ADP-induced, a decrease in the endogenous ADP release response, also with a great effect in patients taking prasugrel.

Dental Caries a Risk Factor for Intracerebral Hemorrhage

Introduction: Streptococcus mutans is a known cause of dental caries that contains a collagen-binding protein, Cnm, and exhibits inhibition of platelet aggregation and matrix metalloproteinase-9 activation. This strain has been linked to aggravation of experimental intracerebral hemorrhage (ICH) and may be a risk factor for ICH. The purpose of this study was to test the association between dental caries and incident ICH. Methods: The presence of dental caries and periodontal disease was assessed in subjects from the Dental Atherosclerosis Risk in Communities (DARIC) study without prior stroke or ICH. This cohort was followed for incident ICH over a period of 10 years. Cox regression was used to compute crude and adjusted hazards ratio from the dental assessment. Results: Among 6,315 subjects, dental surface caries and/or root caries were recorded in 1,338 (27%) subjects. Of those, 7 (0.5%) had incident ICH over a period of 10 years following the visit 4 assessment. Of the remaining 4,977 subjects, 10 (0.2%) had incident ICH. Those with dental caries versus those without dental caries were slightly younger (mean age 62.0 ± 5.7 vs. 62.4 ± 5.6, p = 0.012), had a greater proportion of males (51 vs. 44%, p < 0.001), African Americans (44 vs. 10%, p < 0.001), and were hypertensive (42 vs. 31%, p < 0.001). The association between caries and ICH was significant (crude HR 2.69, 95% CI 1.02–7.06) and strengthened after adjustment for age, gender, race, education level, hypertension, and periodontal disease (adjusted HR 3.88, 95% CI 1.34–11.24). Conclusion: Dental caries is a potential risk for incident ICH after caries detection. Future studies are needed to determine if treatment of dental caries can reduce the risk of ICH.

Platelets and the Lectin Pathway of Complement Activation in Patients with Systemic Lupus Erythematosus or Antiphospholipid Syndrome.

Background Patients with systemic lupus erythematosus (SLE) have an increased risk of thrombosis even when they do not have antiphospholipid syndrome (APS). Interactions between complement activation and activated platelets have been suggested in SLE and APS and could play a role in the increased thrombosis risk. Objectives To explore factors potentially related to the prothrombotic pathophysiology in patients with SLE, primary APS, and healthy controls, by investigating lectin pathway proteins (LPPs), complement activation, platelet aggregation, and platelet activation. Methods This cross-sectional cohort study included 20 SLE patients, 17 primary APS, and 39 healthy controls. Flow cytometry and light transmission aggregometry were used to assess platelet activation and aggregation. Using time-resolved immunofluorometric assays, the plasma concentrations of 11 LPPs and C3dg, reflecting complement activation, were measured. Results H-ficolin plasma concentrations were higher in SLE and APS patients than in controls ( p = 0.01 and p = 0.03). M-ficolin was lower in SLE than in APS ( p = 0.01) and controls ( p = 0.03). MAp19 was higher in APS patients than in SLE patients ( p = 0.01) and controls ( p < 0.001). In APS patients, MASP-2 and C3dg correlated negatively with platelet activation. Platelet-bound fibrinogen after agonist stimulation and C3dg concentrations correlated negatively with platelet activation. Conclusion We observed significant differences between SLE and APS patients regarding complement proteins and platelet activation. Particularly the negative correlations between MASP-2 and C3dg with platelet activation only observed in APS patients suggest that interactions between complement activation and platelets differ in SLE and APS.

Morphofunctional state of platelets in their concentrates depending on storage time

Background.Evaluation of the functional state of platelets in the composition of concentrates is necessary to improve the methods of their production, optimize storage conditions and terms, increase therapeutic efficacy, and reduce the risk of transfusion complications.&#x0D; Aim.Comprehensive study of the morphofunctional state of platelets during the storage of their concentrates.&#x0D; Material and methods.202samples of human platelet apheresis concentrates were studied for storage periods of up to 17days at a temperature of 2224C. The functional state of platelets was assessed using flow cytometry for the expression of P-selectin, active IIb3 integrin, and phosphatidylserines before and after biochemical stimulation. In addition, the mitochondrial potential of platelets, the concentration of intracellular adenosine triphosphate, contractile properties, induced aggregation, and morphological characteristics were studied according to scanning electron microscopy. Statistical analysis was performed by analysis of variance (ANOVA or KruskalWallis).&#x0D; Results.In non-activated platelets during storage, the spontaneous expression of P-selectin increased by an average of 7times, and phosphatidylserines by 3times. The induced expression of P-selectin, integrin IIb3 and phosphatidylserines was the highest on the day of receiving the concentrate and gradually decreased during storage to 1/2of the initial level. Platelet aggregation activity in response to collagen stimulation progressively decreased by an average of 100times, in response to the TRAP peptide by 1.5times. In contrast to the expression of activation markers, the ability of platelets to compress plasma clots in most samples changed slightly, within a few percent; in single samples, the contractile function dropped to zero by 47days of storage, which was combined with platelet hyperactivation and depletion. The mitochondrial potential of platelets and the content of adenosine triphosphate were relatively constant. Morphologically, discoid platelets predominated, however, starting from the 1st day of storage, spindle-shaped platelets, and from the 3rd day spherical forms and microaggregates of platelets accumulated.&#x0D; Conclusion.Platelets in the composition of concentrates initially have a high functional potential, which gradually decreases due to progressive spontaneous activation with a simultaneous decrease in reactivity and structural changes.

Exostosin 1 Knockdown Induces Chemoresistance in MV3 Melanoma Cells by Upregulating JNK and MEK/ERK Signaling.

Heparan sulfate proteoglycans (HSPGs) possess various functions driving malignancy of tumors. However, their impact on tumor cell sensitivity to cytotoxic treatment is far less understood. Aiming to investigate this, we depleted HSPGs by downregulating Exostosin 1 (EXT1), a key enzyme in HS formation, or upregulating heparanase in human MV3 human melanoma cells, and investigated their response to cytotoxic drugs. Cytotoxicity of trametinib, doxorubicin, and mitoxantrone was detected by MTT assay. Insights into intracellular signaling was provided by kinome protein profiler array, and selected kinases were inhibited to investigate their impact on cell sensitization and migratory dynamics. EXT1 knockdown (EXT1kd) in MV3 cells affected the activity of doxorubicin and mitoxantrone, significantly increasing EC50 values two- or fourfold, respectively. Resistance formation was scarcely related to HSPG deficiency, suggested by enzymatic cleavage of HSPG in control cells. Notably, EXT1kd induced an upregulation of EGFR signaling via JNK and MEK/ERK, and hence blocking these kinases returned resistance to a sensitive level. JNK appeared as a key signal component, also inducing higher migratory activity of EXT1kd cells. Furthermore, EXT1kd upregulated thrombotic properties of MV3 cells, indicated by tissue factor and PAR-1 expression, functionally reflected by a stronger activation of platelet aggregation. EXT1 was confirmed to act as a tumor suppressor, shown here for the first time to affect chemosensitivity of melanoma cells.

Plant-food-derived Bioactives in Managing Hypertension: From Current Findings to Upcoming Effective Pharmacotherapies.

A sedentary lifestyle has evoked a high risk of cardiovascular (CV) disease, diabetes, and obesity, all of them with high morbimortality rates and with a common denominator, hypertension. Numerous pharmacological drugs have been used for the treatment of hypertension. However, the side effects associated with the use of existing pharmacological therapies have triggered a demand for plant-based medications. In this connection, the aim of this review was to provide an in-depth analysis of the use of plant-derived bioactives for the effective management of hypertension. Phytoconstituents from leaves, bark, stem, roots, seeds, and fruits of medicinal plants grown in our different regions of the globe have been highly searched. Among them, polyphenols (e.g., flavonoids as quercetin, anthocyanins as cyanidin, tannins as ellagic acid, stilbenes as resveratrol, lignans as honokiol and others as hydroxytyrosol or curcumin), organosulfur compounds (e.g. s-allyl cysteine and allicin), fatty acids (e.g. α-lipoic acid, DHA and oleic acid), alkaloids (e.g. berberine or tetrandrine) and some terpenes have been intensively investigated for the management of hypertension, with effective ability being stated in controlling high blood pressure and related health problems both in vivo and in vitro studies. Some of the activities presented by these bioactive compounds are reducing oxidative stress, renin-angiotensin system control, SIRT1 activation, regulating platelet aggregation and COX activity, anti-atherogenic effects, anti-inflammatory properties, vasorelaxation and other results that translate into the prevention or control of hypertension. The knowledge of these bioactive compounds is important in developing countries where traditional medicine is the majority, but it can also give rise to new approaches in hypertension therapy.

Гомоцистеїн як біомаркер судинної патології

The purpose of the study was to analyze professional literature sources devoted to the influence of homocysteine on the development of vascular disorders. The role of homocysteine as a biomarker of vascular pathology is considered. Materials and methods. Bibliosemantic and analytical methods were used in the study. The analysis of the specialized literature (47 sources) was carried out using information in the scientific-metric databases MEDLINE/PubMed, Scopus and Google Scholar. Results and discussion. Homocysteinemia is a pathological condition associated with quite serious consequences for the body; the endothelium of vessels suffers mostly: systemic endothelial dysfunction develops (including disruption of the synthesis and exchange of the endothelium-relaxing factor – nitric oxide), oxidative stress, activation of platelet aggregation, hypercoagulation occurs (due to a decrease in the activity of heparin, thrombomodulin and an increase in the activity of thromboxane A2). The prevalence of hyperhomocysteinemia in Ukraine is quite high and reaches 10% among healthy adults, 2% among young people and adolescents, from 13% to 43% in patients with cardiovascular pathology. The C677T polymorphism of the methylenetetrahydrofolate reductase gene is widespread among the population of Ukraine (40.7%). Researchers believe that hyperhomocysteinemia is an independent and modifiable risk factor for vascular pathology. Conclusion. Homocysteine is recognized as one of the markers of vascular pathology. Scientists consider the activation of oxidative stress, damage to the endothelium, stimulation of the proliferation of smooth muscle cells, and pro-inflammatory effects to be the main mechanisms of vascular damage in hyperhomocysteinemia. The described pathological changes are accompanied by a violation of the regulation of vascular tone, mostly due to a decrease in the synthesis of nitric oxide. Unbalanced synthesis of nitric oxide causes and potentiates oxidative stress, the processes of atherothrombogenesis. The professional literature presents enough scientific data that convincingly prove the role of homocysteine in the development of cardiovascular and neurological pathology. However, there is a small number of studies devoted to the relationship between hyperhomocysteinemia and periodontal diseases, and the pathogenetic mechanisms of the influence of hyperhomocysteinemia on the development of vascular disorders in coronavirus disease are not fully disclosed

Clinical and diagnostic significance of plasminogen activator inhibitor type 1 in the early development of coronary heart disease

Aim to determine the quantitative content of the tissue plasminogen activator inhibitor type 1 (PAI-1) and to determine its relationship with risk factors for coronary artery disease.&#x0D; Material and methods. The study included 67 male and female patients (36 men and 31 women) with coronary artery disease (CAD) and stable angina classes II-IV with the presence of hypertension. The mean age of the patients was 60.20.76 years. In all patients, clinical and anamnestic data were collected during the examination. For pain assessment, we used a verbal scale. We registered the anthropometric data and body mass index. Biochemical and instrumental methods of research were used according to the standards for diagnosing CAD. We assessed the occurrence of risk factors in patients included in the study. A complex of general clinical laboratory and instrumental tests was used, as well as the method of enzyme immunoassay ELISA, which determined the level of PAI-1, platelet aggregation activity, renin, aldosterone, cortisol and endothelin.&#x0D; Results. The group of patients demonstrated a reliable elevation of PAI-1 plasma level. Compared with practically healthy people, the level of this biomarker was 1.6 times higher in patients with coronary artery disease. A significant elevation of PAI-1 level indicates an increased risk of endothelial and hemostasiological disorders in patients, which, in turn, increases the risk of thrombogenic complications. The study revealed the correlation of PAI-1 protein in CAD patients with such risk factors as mixed anxiety-depressive disorder, smoking, hypertension, obesity, and hypercholesterolemia.