Utility of thromboelastography with platelet mapping (TEG-PM) for monitoring platelet transfusion in qualitative platelet disorders.

BACKGROUND: Patients with pathogenic variants in RASGRP2 (inherited platelet disorder (IPD)-18) have normal platelet counts but show impaired platelet aggregation due to diminished activation of αIIbß3 integrin. This defect results in moderate to severe bleeding episodes, especially following surgical procedures, which require patients to be transfused with platelets and/or pro-hemostatic agents. We recently demonstrated that hemostatic efficacy of transfused platelets is limited by dysfunctional endogenous platelets in a mouse model of IPD-18 (Rasgrp2-/- mice), as dysfunctional platelets were recruited to the forming hemostatic plug but did not participate in clot contraction. Consequently, higher amounts of transfused platelets were required to outcompete these dysfunctional cells and to reverse bleeding. OBJECTIVE: We here studied the usefulness of thromboelastography with platelet mapping (TEG-PM), a method to evaluate platelet-dependent clot contraction, for ex vivo monitoring of the hemostatic potential in Rasgrp2-/- mice transfused with various amounts of wild-type (WT) platelets. RESULTS: Rasgrp2-/- whole blood samples did not contract in TEG-PM, consistent with a critical role of this protein in αIIbß3 activation. Addition of WT platelets improved TEG parameters (K time, α-angle, MA) in a ratio dependent manner, consistent with our recent in vivo studies showing impaired hemostasis at a 5:1, but not at a 2:1 ratio of mutant to WT platelets. K and α values were identified as better predictors of transfusion efficacy than MA, the most platelet-dependent TEG parameter. CONCLUSION: This proof-of-concept study supports the use of TEG-PM to monitor platelet transfusion ratios and hemostatic potential in IPD-18 and potentially other platelet disorders.

New Findings Regarding the Effects of Selected Blue Food Colorants (Genipin, Patent Blue V, and Brilliant Blue FCF) on the Hemostatic Properties of Blood Components In Vitro.

Natural and synthetic colorants present in food can modulate hemostasis, which includes the coagulation process and blood platelet activation. Some colorants have cardioprotective activity as well. However, the effect of genipin (a natural blue colorant) and synthetic blue colorants (including patent blue V and brilliant blue FCF) on hemostasis is not clear. In this study, we aimed to investigate the effects of three blue colorants-genipin, patent blue V, and brilliant blue FCF-on selected parameters of hemostasis in vitro. The anti- or pro-coagulant potential was assessed in human plasma by measuring the following coagulation times: thrombin time (TT), prothrombin time (PT), and activated partial thromboplastin time (APTT). Moreover, we used the Total Thrombus formation Analysis System (T-TAS, PL-chip) to evaluate the anti-platelet potential of the colorants in whole blood. We also measured their effect on the adhesion of washed blood platelets to fibrinogen and collagen. Lastly, the cytotoxicity of the colorants against blood platelets was assessed based on the activity of extracellular lactate dehydrogenase (LDH). We observed that genipin (at all concentrations (1-200 µM)) did not have a significant effect on the coagulation times (PT, APTT, and TT). However, genipin at the highest concentration (200 µM) and patent blue V at the concentrations of 1 and 10 µM significantly prolonged the time of occlusion measured using the T-TAS, which demonstrated their anti-platelet activity. We also observed that genipin decreased the adhesion of platelets to fibrinogen and collagen. Only patent blue V and brilliant blue FCF significantly shortened the APTT (at the concentration of 10 µM) and TT (at concentrations of 1 and 10 µM), demonstrating pro-coagulant activity. These synthetic blue colorants also modulated the process of human blood platelet adhesion, stimulating the adhesion to fibrinogen and inhibiting the adhesion to collagen. The results demonstrate that genipin is not toxic. In addition, because of its ability to reduce blood platelet activation, genipin holds promise as a novel and valuable agent that improves the health of the cardiovascular system and reduces the risk of cardiovascular diseases. However, the mechanism of its anti-platelet activity remains unclear and requires further studies. Its in vivo activity and interaction with various anti-coagulant and anti-thrombotic drugs, including aspirin and its derivatives, should be examined as well.

CORRELATION OF NEUTROPHIL-LYMPHOCYTE RATIO (NLR) AND PLATELET-LYMPHOCYTE RATIO (PLR) WITH COMPLICATIONS AND POSTOPERATIVE SURVIVAL IN SURGERY FOR BONE METASTASIS OF THE APPENDICULAR SKELETON / CORRELAÇÃO DO NEUTROPHIL-LYMPHOCYTE RATIO (NLR) E PLATELET-LYMPHOCYTE RATIO (PLR) COM COMPLICAÇÕES E SOBREVIDA PÓS-OPERATÓRIAS NA CIRURGIA PARA METÁSTASE ÓSSEA DO ESQUELETO APENDICULAR

Introduction: Bone metastasis is the most common malignant neoplasm of the skeleton, and surgical decision-making depends on multiple factors, including postoperative complications and life expectancy. The identification of new prognostic factors can assist in decision making. Objective: In long bones metastases, to analyze the incidence of complications and postoperative survival up to 1 year, correlating them with NLR and PLR. Method: Review of 160 medical records of patients who underwent surgery for bone metastasis in the appendicular skeleton. In addition to epidemiological characteristics, NLR and PLR values were determined, correlating them with survival and complications. Result: Women represented 64.5% with a primary breast tumor in 62.6%; the proximal femur was the most affected; median survival was 13.2 months and in 1 year 34.7%. Tumor resection with endoprosthesis was more common. The post-surgical complication rate was 10% and the average time for post-operative complications to occur was 27.9 days (0-140). An association between the neutrophil variable and postoperative complications was found (p=0.04). For every 100 more units of neutrophils there was a 1% increase in the chances of post-surgical complications. Mean NLR and PLR values were, respectively, 5.3 (0.2-30.7) and 199.7 (32.1-676.7). Patients with NLR NLR ≥ 2 (p<0,001) showed a decrease in survival from 92,3% to 62,5% at the 3rd month, and from 61,5% to 31,3% at 1 year. Those with PLR ≥209 (p<0.001) showed a decrease in survival from 69% to 59.3% at the 3rd month, and from 40.2% to 25.9% at 1 year. Conclusion: There was no positive association between NLR and PLR with postoperative complications, but strongly yes with survival from the 3rd month after surgery.

Introdução: Metástase óssea é a neoplasia maligna mais comum do esqueleto, e a tomada de decisão cirúrgica depende de múltiplos fatores, incluindo as complicações pós-operatórias e a expectativa de vida. A identificação de novos fatores prognósticos pode auxiliar na tomada de decisão. Objetivo: Analisar em metástases de ossos longos, a incidência de complicações e sobrevida pós-operatórias até 1 ano correlacionando-as com NLR e PLR. Método: Revisão de 160 prontuários de operados por metástase óssea no esqueleto apendicular. Além de características epidemiológicas, foram determinados os valores de NLR e PLR correlacionando-os com sobrevida e complicações. Resultado: Mulheres representaram 64,5% com tumor primário na mama em 62,6%; o fêmur proximal foi o mais acometido; sobrevida média foi 13,2 meses e a de 1 ano 34,7%; ressecção tumoral com endoprótese foi mais comum. A taxa de complicação pós-cirúrgicas foi de 10% e o tempo médio para a ocorrência de complicações pós-operatórias foi de 27,9 dias (0-140). Foi encontrada associação da variável neutrófilos com a complicação pós-operatória (p=0,04). A cada 100 unidades a mais de neutrófilos houve aumento de 1% nas chances de complicações pós-cirúrgicas. Valores médios do NLR e PLR foram, respectivamente, 5,3 (0,2-30,7) e 199,7 (32,1-676,7).  Os pacientes com NLR ≥ 2 (p<0,001) apresentaram diminuição na sobrevida de 92,3% para 62,5% no 3° mês e de 61,5% para 31,3% em 1 ano. Aqueles com PLR ≥209 (p<0,001) apresentaram diminuição na sobrevida de 69% para 59,3% no 3° mês, e de 40,2% para 25,9% em 1 ano. Conclusão: Não foi verificada associação positiva entre o NLR e o PLR com complicações pós-operatórias, mas com sobrevida fortemente sim, a partir do 3° mês de pós-operatório.

CAN NEUTROPHIL-LYMPHOCYTE RATIO (NLR) AND PLATELET-LYMPHOCYTE RATIO (PLR) BE PROGNOSTIC INDICATORS FOR OPERATIVE COMPLICATIONS AND SURVIVAL IN BONE METASTASES? / NEUTROPHIL-LYMPHOCYTE RATIO (NLR) E PLATELET-LYMPHOCYTE RATIO (PLR) PODEM SER INDICADORES DE PROGNÓSTICO PARA COMPLICAÇÕES OPERATÓRIAS E SOBREVIDA EM METÁSTASES ÓSSEAS?

Introduction: In general, inflammation stimulates the production and release of neutrophils and, at the same time, decreases the production of lymphocytes. Lymphopenia reflects that cell-mediated immunity is impaired, while neutrophilia represents a response to systemic inflammation in these cancers. Objective: To review the incidence of complications and postoperative survival rates in patients with bone metastases in long bones, correlating them with markers NLR and PLR. Method: Narrative review carried out collecting information published on virtual platforms in Portuguese and English, initially carried out by searching for descriptors related to the topic, which were: "lower extremity, surgery, metastasis, epidemiology, postoperative complications, neutrophils, lymphocytes, platelets". The extension incorporated AND or OR, by title and/or summary, and full reading of the texts most related to the topic. Result: 21 articles were included. Conclusion: The higher both the NLR and PLR are associated with lower survival in patients with bone metastases when undergoing surgical treatment, especially after 3 months postoperatively. However, there is still no confirmation that they signal any outcome, favorable or not, in relation to postoperative complications.

Introdução : De um modo geral, a inflamação estimula a produção e liberação de neutrófilos e, ao mesmo tempo, diminui a produção de linfócitos. A linfopenia reflete que a imunidade mediada pelas células é prejudicada, enquanto a neutrofilia representa resposta à inflamação sistêmica nesses cânceres. Objetivo : Revisar nos pacientes com metástase óssea em ossos ao longo da incidência de complicações e taxas de sobrevida pós-operatória correlacionando-as com os marcadores NLR e PLR. Método : Revisão narrativa feita colhendo informações publicadas em plataformas virtuais em português e inglês inicialmente realizada por busca dos descritores relacionados ao tema que foram: "extremidade inferior, cirurgia, metástase, epidemiologia, complicações pós-operatórias, neutrófilos, linfócitos, plaquetas" e seus equivalentes em inglês " extremidade inferior, cirurgia, metástase, epidemiologia, sobrevivência, complicações, neutrófilos, linfócitos, plaquetas sanguíneas ". A extensão incorporou AND ou OR, pelo título e/ou resumo, e leitura na íntegra dos textos mais relacionados ao tema.  Resultado : Foram incluídos 21 artigos. Conclusão : Quanto maiores, tanto o NLR quanto o PLR estão associados à menor sobrevida em pacientes com MO quando submetidos ao tratamento cirúrgico, especialmente após 3 meses de pós-operatório. Contudo, ainda não há confirmação de que eles sinalizam algum estágio, positivo ou não, em relação às complicações pós-operatórias.

Investigating the role of rotenone on human blood platelets: Molecular insights into abnormal platelet functions in Parkinson's disease.

Parkinson's disease (PD) is a predominant neuromotor disorder characterized by the selective death of dopaminergic neurons in the midbrain. The majority of PD cases are sporadic or idiopathic, with environmental toxins and pollutants potentially contributing to its development or exacerbation. However, clinical PD patients are often associated with a reduced stroke frequency, where circulating blood platelets are indispensable. Although platelet structural impairment is evident in PD, the platelet functional alterations and their underlying molecular mechanisms are still obscure. Therefore, we investigated rotenone (ROT), an environmental neurotoxin that selectively destroys dopaminergic neurons mimicking PD, on human blood platelets to explore its impact on platelet functions, thus replicating PD conditions in vitro. Our study deciphered that ROT decreased thrombin-induced platelet functions, including adhesion, activation, secretion, and aggregation in human blood platelets. As ROT is primarily responsible for generating intracellular reactive oxygen species (ROS), and ROS is a key player regulating the platelet functional parameters, we went on to check the effect of ROT on platelet ROS production. In our investigation, it became evident that ROT treatment resulted in the stimulation of ROS production in human blood platelets. Additionally, we discovered that ROT induced ROS production by augmenting Ca2+ mobilization from inositol 1,4,5-trisphosphate receptor. Apart from this, the treatment of ROT triggers protein kinase C associated NADPH oxidase-mediated ROS production in platelets. In summary, this research, for the first time, highlights ROT-induced abnormal platelet functions and may provide a mechanistic insight into the altered platelet activities observed in PD patients.

Should HLA and HPA-matched platelet transfusions for patients with Glanzmann Thrombasthenia or Bernard-Soulier syndrome be standardized care? A Dutch survey and recommendations.

BACKGROUND: Glanzmann thrombasthenia (GT) and Bernard-Soulier syndrome (BSS) patients require frequent platelet transfusions and hence have an increased risk for alloimmunization against donor Human Leukocyte Antigens (HLA) when no HLA-matching is performed. Knowing that Human Platelet Antigens (HPA) are located on the platelet glycoproteins that can be absent in these patients, preventive HPA-matching may also be considered. Uniform recommendations on this topic lack in transfusion guidelines making standard practice unclear, therefore, we aimed to provide a framework for matched platelet transfusions. STUDY DESIGN AND METHODS: We conducted a targeted literature search and a national survey of Dutch (pediatric) hematologists from July to September 2021. RESULTS: We found 20 articles describing platelet transfusion policies in 483 GT-patients and 29 BSS-patients, both adults and children. Twenty surveys were returned for full analysis. All responders treated patients with platelet disorders, including GT (n = 36 reported) and BSS (n = 29 reported). Of respondents, 75% estimated the risk of antibody formation as "likely" for HLA and 65% for HPA. Formation of HLA antibodies was reported in 5 GT and in 5 BSS-patients, including one child. Fifteen respondents gave preventive HLA-matched platelets in elective setting (75%). Three respondents additionally matched for HPA in GT-patients (15%). Main argument for matched platelet transfusions was preventing alloimmunization to safeguard the effectivity of 'random' donor-platelets in acute settings. CONCLUSION: Elective HLA-matching for GT and BSS-patients is already conducted by most Dutch (pediatric) hematologists. HPA-matching is mainly applied when HPA-antibodies are formed. Based on the current literature and the survey, recommendations are proposed.

Use of crushed tranexamic acid tablets in water for paediatric patients with bleeding disorders.

BACKGROUND: Ε-Aminocaproic acid oral solution (EACA OS) is the only commercially available antifibrinolytic for patients who cannot swallow tablets. Insurance denials and high costs remain barriers to its use. OBJECTIVES: To determine the safety and efficacy of crushed tranexamic acid tablets in water (cTXAw) for children with bleeding disorders. METHODS: We retrospectively reviewed records of children (<10 years) with bleeding disorders who received cTXAw or EACA OS from 1 December 2018, through 31 July 2022, at Mayo Clinic (Rochester, Minnesota). Bleeding outcomes were defined according to ISTH criteria. RESULTS: Thirty-two patients were included (median age, 3 years; male, n = 23). Diagnoses were VWD (n = 17), haemophilia (n = 5), FVII deficiency (n = 3), inherited platelet disorder (n = 4), ITP (n = 2), and combined FV and FVII deficiencies (n = 1). Thirty-two courses of cTXAw (monotherapy 24/32; mean duration 6 days) and fifteen courses of EACA (monotherapy 12/15; mean duration 5 days) were administered. No surgical procedures (n = 28) were complicated by bleeding. Of the 19 bleeding events, 16 had effective haemostasis, two had no reported outcome, and one had no response. cTXAw and EACA were equally effective in preventing and treating bleeding (p value > .1). No patients had adverse effects. Eight of 19 patients (42%) who were initially prescribed EACA OS did not receive it because of cost or insurance denial. The estimated average wholesale price of one treatment was $94 for cTXAw and $905 for EACA OS. CONCLUSIONS: CTXAw appears to be an effective, safe, and low-cost alternative option to EACA OS for young children with bleeding disorders.

ISTH bleeding assessment tool and platelet function analyzer in children with mild inherited platelet function disorders.

OBJECTIVES: To evaluate the diagnostic performance of platelet function analyzer (PFA) and The International Society on Thrombosis and Hemostasis bleeding-assessment-tool (ISTH-BAT) in detecting mild inherited platelet function disorders (IPFDs) in children with suspected bleeding disorders. METHODS: Prospective single-center diagnostic study including consecutive patients <18 years with suspected bleeding disorder and performing a standardized workup for platelet function defects including ISTH-BAT, PFA, platelet aggregation testing, blood smear-based immunofluorescence, and next-generation sequencing-based genetic screening for IPFDs. RESULTS: We studied 97 patients, of which 34 von Willebrand disease (VWD, 22 type-1, 11 type-2), 29 IPFDs (including delta-/alpha-storage pool disease, Glanzmann thrombasthenia, Hermansky-Pudlak syndrome) and 34 with no diagnosis. In a model combining PFA-adenosine diphosphate (ADP), PFA-epinephrine (EPI), and ISTH-BAT overall performance to diagnose IPFDs was low with area under the curves of 0.56 (95% CI 0.44, 0.69) compared with 0.84 (95% CI 0.76, 0.92) for VWD. Correlation of PFA-EPI/-ADP and ISTH-BAT was low with 0.25/0.39 Spearman's correlation coefficients. PFA were significantly prolonged in patients with VWD and Glanzmann thrombasthenia. ISTH-BAT-scores were only positive in severe bleeding disorders, but not in children with mild IPFDs or VWD. CONCLUSION: Neither ISTH-BAT nor PFA or the combination of both help diagnosing mild IPFDs in children. PFA is suited to exclude severe IPFDs or VWD and is in this regard superior to ISTH-BAT in children.

Potential for modulation of platelet function via adenosine receptors during inflammation.

Traditionally, platelets are known to play an important role in haemostasis and thrombosis; however, they serve also as important modulators of inflammation and immunity. Platelets secrete adhesion molecules and cytokines, interact with leukocytes and endothelium, and express toll-like receptors involved in a direct interaction with pathogens. Platelets express A2A and A2B subtypes of receptors for adenosine. The activation of these receptors leads to an increase in cAMP concentration in the cytoplasm, thereby resulting in inhibited secretion of pro-inflammatory mediators and reduced cell activation. Therefore, platelet adenosine receptors could be a potential target for inhibiting platelet activation and thus down-regulating inflammation or immunity. The biological effects of adenosine are short-lasting, because the compound is rapidly metabolized; hence, its lability has triggered efforts to synthesize new, longer-lasting adenosine analogues. In this article, we have reviewed the literature regarding the pharmacological potential of adenosine and other agonists of A2A and A2B receptors to affect platelet function during inflammation. LINKED ARTICLES: This article is part of a themed issue on Platelet purinergic receptor and non-thrombotic disease. To view the other articles in this section visit http://onlinelibrary.wiley.com/doi/10.1111/bph.v181.4/issuetoc.

Immune Thrombocytopenic Purpura in an Adult Male: A Case Report.

Immune thrombocytopenic purpura (ITP) is an autoimmune disease characterized by immune-mediated destruction of platelets, resulting in a decreased blood platelet count (less than 100 x 109/L) in the absence of other known etiology of thrombocytopenia. ITP is uncommon in adult males. The signs and symptoms of ITP vary widely and are quite diverse. The degree of thrombocytopenia and bleeding are not always correlated. Timely diagnosis, intervention, and regular monitoring can easily prevent complications. We report a case of a 22-year-old male presented with gum bleeding along with purpura and ecchymosis over the upper limb, lower limb, trunk, and face.

Diversity of Megakaryocytes.

Megakaryocytes are commonly known as large, polyploid, bone marrow resident cells that contribute to hemostasis through the production of platelets. Soon after their discovery in the 19th century, megakaryocytes were described in tissue locations other than the bone marrow, specifically in the lungs and the blood circulation. However, the localization of megakaryocytes in the lungs and the contribution of lung megakaryocytes to the general platelet pool has only recently been appreciated. Moreover, the conception of megakaryocytes as uniform cells with the sole purpose of platelet production has been challenged. Here, we review the literature on megakaryocyte cell identity and location with a special focus on recent observations of megakaryocyte subpopulations identified by transcriptomic analyses.

Innovative i-PRF semisurgical method for gingival augmentation and root coverage in thin periodontal phenotypes: a preliminary study.

OBJECTIVES: The aim of the study was to evaluate the effect of injectable platelet-rich fibrin (i-PRF) on gingival thickness and gingival recession in individuals with thin periodontal phenotypes. METHOD AND MATERIALS: In this prospective study, i-PRF was applied via a semisurgical method to augment 53 tooth regions with thin periodontal phenotypes. In order to ensure that sufficient blood clot formed on the side of the gingiva facing the bone and that i-PRF reached the area, a minimal incision was made with the help of a scalpel in the apical region of the relevant region, and the periosteum was elevated with a microsurgical instrument. To ensure sustained exposure to angiogenetic growth factors and enhance the histoconductive properties, i-PRF injection was applied to the relevant areas in four sessions at 10-day intervals. RESULTS: An increase in gingival thickness was achieved in 92.5% of the areas treated with i-PRF, and the desired gingival thickness (0.8 mm) was achieved in 44.9% of these areas. In addition, significant reductions in the amount of recession were observed in 83.3% of the 12 gingival recession areas (P = .005). Moreover, complete coverage was achieved in 60% of these regions. CONCLUSION: With the new i-PRF semisurgical method, it was shown that gingival thickness can be increased in tooth regions with thin gingiva, and that areas of gingival recession can be covered. Further comprehensive studies are needed to fully understand the role of i-PRF in enhancing angiogenesis and the histoconductive properties of this fully autogenous blood concentrate.

Multicolor flow cytometry in clinical samples for platelet signaling assessment.

Background: Availability of multichannel cytometers and specific commercial antibodies makes flow cytometry a new option to simultaneously assess multiple intracellular platelet signaling pathways for clinical purposes, in small volume of blood or low platelet count. Objectives: To describe a multicolor flow cytometry with fluorescent barcoding technique for screening signaling pathways downstream membrane receptors of major platelet agonists (adenosine diphosphate, thrombin, thromboxane, and collagen). Methods: By comparison with immunoblotting, we first selected the target phosphoproteins, AKT, P38MAPK, LIMK, and SPL76; the times of stimulation; and phosphoflow barcoding conditions. We then performed a clinical study on whole blood of patients without evidence of blood platelet disorder on standard biological screening, consulting for trivial or occasionally provoked bleeds without familial antecedent (bleeding of unknown origin, n = 23) or type-1 von Willebrand disease (n = 9). In addition, we included a small group of patients with definite platelet disorders (Glanzmann thrombasthenia, δ-storage pool deficiency, and immune glycoprotein VI-related disease with granule secretion defect). Results: The range, kinetics, and distribution of fluorescence intensity were established for each agonist-target protein combination. Principal component analysis indicates a correlation in response to a target phosphoprotein (AKT and P38MAPK) to different agonists but no correlation in the response of different target phosphoproteins to the same agonist. The heterogeneity of individual responses in the whole population displayed was analyzed using clustering algorithm. Patients with platelet storage pool deficiency were positioned as lowest responders on the heatmap. Conclusion: In complement of functional tests, this study introduces a new approach for rapid platelet signaling profiling in clinical practice.

Platelet to lymphocyte and neutrophil to lymphocyte ratio in the first trimester of pregnancy, are they useful for predicting spontaneous miscarriage? A case-control study.

Background: In 15% of all clinical pregnancies, a miscarriage can occur, but the exact cause of this phenomenon is not fully understood. However, it is believed that a faulty placenta, which triggers an inflammatory response in the mother's body, may be one of the causes. Medical literature has increasingly focused on 2 indicators of inflammation, the platelet-lymphocyte ratio (PLR) and neutrophil-lymphocyte ratio (NLR). Despite this, there has yet to be a study conducted that examines the rates of PLR and NLR in cases of miscarriage. Objective: This study aims to determine whether there is an increase in complete blood count inflammatory parameters such as NLR and PLR in women who experience miscarriages. Materials and Methods: This retrospective case-control study was conducted from March 2021 to March 2022, across 3 academic hospitals in Tehran, Iran. A total of 240 participants were enrolled comprising individuals with either miscarriages or normal pregnancies (n = 120/each). Data were collected from the medical records of participants aged between 18-42 yr old, with gestational age ranging from 6-13 wk. The demographic information, including age, body mass index, parity, history of abortion, number of abortions, number of living children, hematocrit and hemoglobin levels, platelet distribution width (PDW), PLR, NLR, mean platelet volume, and platelet were extracted from their records. The gestational age was also recorded. Results: A total of 240 participants were recruited for the study. PDW, NLR, PLR, and lymphocyte values were higher in the miscarriage group compared to the healthy normal pregnant women (p < 0.001). Mean platelet volumes were found to be lower in the miscarriage group compared to the healthy normal pregnant women (p < 0.001). Conclusion: Although, no statistically significant difference was observed in the hemoglobin, hematocrit, platelets, and neutrophils in these 2 groups of pregnant women. The higher inflammatory markers including PDW, NLR, and PLR could potentially aid in the speculation of defective placentation as a contributing factor to the development of miscarriage. Measurement of these markers may be useful to predict pregnancy leading to miscarriage.

Prothrombin consumption as an indicator of hemorrhagic phenotype in mild platelet function disorders.

BACKGROUND: The bleeding risk of patients with mild platelet function disorders is difficult to assess and their phenotype remains ill-explored. AIM: This study was designed to establish a comprehensive biological phenotype of patients with mild platelet function disorders. METHODS: Twenty patients were included with persistent abnormal light transmission aggregometry (LTA). The ISTH bleeding assessment tool (ISTH-BAT) was assessed to identify laboratory analyses associated with an abnormal hemorrhagic score. RESULTS: The majority of patients had defects that might affect Gαi protein signaling pathways or minor abnormalities. No LTA nor flow cytometry parameters were associated with an above-normal hemorrhagic score. However, prothrombin consumption, which corresponds to the ratio of serum residual factor II to plasma residual factor II, was significantly higher (p = .006) in the abnormal ISTH-BAT group (mean = 14%, SD = 6) compared with the normal ISTH-BAT group (mean = 8%, SD 4). Prothrombin consumption was significantly associated with ISTH-BAT score (r = .5287, IC 95% 0.0986-0.7924, p = .0165). CONCLUSION: In this group of patients, there was an association between a pathological bleeding score and increased prothrombin consumption. This test could be used as an additional indicator of platelet function abnormality liable to be related to bleeding risk.

Clinical Cytometry for Platelets and Platelet Disorders.

Clinical flow cytometry tests for inherited and acquired platelet disorders are useful diagnostic tools but are not widely available. Flow cytometric methods are available to detect inherited glycoprotein deficiencies, granule release (secretion defects), drug-induced thrombocytopenias, presence of antiplatelet antibodies, and pharmacodynamic inhibition by antiplatelet agents. New tests take advantage of advanced multicolor cytometers and allow identification of novel platelet subsets by high-dimensional immunophenotyping. Studies are needed to evaluate the value of these new tests for diagnosis and monitoring of therapy in patients with platelet disorders.

Preoperative Glutamine Supplementation in Gastric Cancer-Thrombocyte Phagocytic Activity and Early Postoperative Outcomes.

BACKGROUND: The aim of this study was to determine the phagocytic activity of thrombocytes in patients with gastric cancer and to assess the effect of oral and parenteral preoperative glutamine-based immunonutrition on nutritional status, thrombocyte phagocytic activity, and early postoperative outcomes. METHODS: Patients suffering from invasive gastric cancer had been treated with preoperative immunonutrition with glutamine, and they were compared to patients without nutritional treatment. Nutritional status, percentage of weight loss, and BMI were assessed. Levels of total protein, albumin, cholesterol, triglycerides, platelets, and their phagocytic ability were measured twice. Postsurgical complications were assessed via the Clavien-Dindo classification. RESULTS: Group I consisted of 20 patients with an oral glutamine-10 g daily. Group II had 38 patients who received intravenous glutamine, 1.5 mL per kg body weight of Dipeptiven. Group III consisted of 25 patients who did not receive preoperative immunonutrition. In total, 47% of patients in Group I, 54% of patients in Group II, and 33% of patients in Group III were malnourished. In Group I, the percentage of phagocytizing platelet (%PhP) was 1.1 preoperatively and 1.2 postoperatively. The phagocytic index (PhI) was 1.0 and 1.1. In Group II, %PhP was 1.1 and 1.2 and PhI was 1.0 and 1.1. In Group III, the %PhP was 1.0 and 1.2 and PhI was 1.0 and 1.1. An increase in triglyceride level was observed in both immunonutrition groups. There was a decline in total protein and albumin level in Group II. In Group III, there was a decline in total protein, albumin, and cholesterol level. The total platelet count and PhI were increased in both immunonutrition groups. There was also a rise in %PhP in Group II. In Group III, there was a rise in blood platelet level, %PhP, and PhI. The complication rates were 53% in Group I, 29% in Group II, and 40% in Group III. CONCLUSIONS: In invasive gastric cancer, laboratory nutritional parameters are significantly reduced, causing malnutrition in 44.7% of patients. Oral glutamine supplementation inhibited the postoperative decline in protein metabolism parameters; however, this did not affect the reduction in the percentage of postoperative complications. Glutamine used preoperatively significantly reduced the percentage of serious surgical complications, regardless of the way it was supplemented. Patients with invasive gastric cancer have a significant decrease in platelet phagocytic activity. The administered preoperative parenteral nutrition and the surgical procedure itself influenced the improvement of the phagocytic activity of blood platelets. Glutamine did not have this effect, regardless of the route of administration.

Response of human blood platelets to preparations from leaves of Paulownia Clon in Vitro 112.

Paulownia Clon in Vitro 112, also called the Oxytree, is a fast-growing hybrid of two trees belonging to the Paulowniaceae family - P. elongata and P. fortunei. It demonstrates a wide range of biological effects (including antioxidant, anti-inflammatory, antibacterial, and neuroprotective) due to the high concentration of secondary metabolites. Our previous results showed an in vitro antioxidant and antiplatelet activity of the extract and four fractions (A-D) from the leaves of Paulownia Clon in Vitro 112 in human plasma and washed blood platelets. Here, we used a microchip flow chamber-based thrombus formation analysis system (T-TAS) and flow cytometry to assess the anticoagulant and antiplatelet activity of the extract and four fractions with different chemical content (A-D) from Paulownia Clon in Vitro 112 leaves in human whole blood. Two tested fractions: fraction C and D (at the concentrations of 5 and 50 µg/mL) inhibited the exposition of the active form of GPIIb/IIIa (integrin αIIbß3) on the surface of blood platelets stimulated by ADP and collagen. The antiplatelet activity of fraction C is likely due to its high verbascoside content and the presence of apigenin's derivatives. Fraction D contains triterpenoids, including ursolic, pomoleic, and maslinic acid, which could be responsible for decreased activation of ADP- and collagen-stimulated blood platelets. These results suggest that fractions C and D might be promising sources of phytochemicals with antiplatelet activity, which are important for prophylaxis and treatment of cardiovascular diseases associated with hyperactivation of blood platelets. However, further research is needed to ascertain which exact compounds and mechanisms are responsible for this phenomenon.

Discrimination Between Pre- and Postcapillary Pulmonary Hypertension Using Platelet RNA.

Background Appropriate treatment of pulmonary hypertension (PH) is critically dependent on accurate discrimination between pre- and postcapillary PH. However, clinical discrimination is challenging and frequently requires a right heart catheterization. Existing risk scores to detect postcapillary PH have suboptimal discriminatory strength. We have previously shown that platelet-derived RNA profiles may have diagnostic value for PH detection. Here, we hypothesize that platelet-derived RNAs can be employed to select unique biomarker panels for the discrimination between pre- and postcapillary PH. Methods and Results Blood platelet RNA from whole blood was isolated and sequenced from 50 patients with precapillary PH (with different PH subtypes) as well as 50 patients with postcapillary PH. RNA panels were calculated by ANOVA statistics, and classifications were performed using a support vector machine algorithm, supported by particle swarm optimization. We identified in total 4279 different RNAs in blood platelets from patients with pre- and postcapillary PH. A particle swarm optimization-selected RNA panel of 1618 distinctive RNAs with differential levels together with a trained support vector machine algorithm accurately discriminated patients with precapillary PH from patients with postcapillary PH with 100% sensitivity, 60% specificity, 80% accuracy, and 0.95 (95% CI, 0.86-1.00) area under the curve in the independent validation series (n=20). Conclusions This proof-of-concept study demonstrates that particle swarm optimization/support vector machine-enhanced classification of platelet RNA panels may be able to discriminate precapillary PH from postcapillary PH. This research provides a foundation for the development of a blood test with a high negative predictive value that would improve early diagnosis of precapillary PH and prevents unnecessary invasive testing in patients with postcapillary PH.

Unveiling the mechanism of platelet dysfunction in Parkinson's disease: The effect of 6-hydroxydopamine on human blood platelets.

INTRODUCTION: Parkinson's disease (PD) is a progressive neuronal illness often linked to increased cardiovascular complications, such as myocardial infarction, cardiomyopathy, congestive heart failure, and coronary heart disease. Platelets, which are the essential components of circulating blood, are considered potential players in regulating these complications, as platelet dysfunction is evident in PD. These tiny blood cell fragments are supposed to play a crucial role in these complications, but the underlying molecular processes are still obscure. METHODS: To gain a better understanding of platelet dysfunction in PD, we investigated the impact of 6-hydroxydopamine (6-OHDA), an analog of dopamine that simulates PD by destroying dopaminergic neurons, on human blood platelets. The levels of intraplatelet reactive oxygen species (ROS) were assessed using H2DCF-DA (20 µM), while mitochondrial ROS was evaluated using MitoSOX™ Red (5 µM), and intracellular Ca2+ was measured with Fluo-4-AM (5 µM). The data were acquired through the use of both a multimode plate reader and a laser-scanning confocal microscope. RESULTS: Our findings showed that 6-OHDA treatment increased the production of ROS in human blood platelets. The increase in ROS was confirmed by the ROS scavenger, NAC, and was also reduced by inhibiting the NOX enzyme with apocynin. Additionally, 6-OHDA potentiated mitochondrial ROS production in platelets. Furthermore, 6-OHDA triggered the intraplatelet Ca2+ elevation. This effect was mitigated by the Ca2+ chelator BAPTA, which decreased the ROS production triggered by 6-OHDA in human blood platelets, while the IP3 receptor blocker, 2-APB, reduced the formation of ROS induced by 6-OHDA. CONCLUSION: Our findings suggest that the 6-OHDA-induced ROS production is regulated by the IP3 receptor-Ca2+-NOX signaling axis in human blood platelets, where the platelet mitochondria also play a significant role. This observation provides a crucial mechanistic understanding of the altered platelet activities that are commonly observed in PD patients.