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BACKGROUND: Exercise-induced pulmonary haemorrhage (EIPH) in athletic horses is characterized by the presence of blood from the lungs in the tracheobronchial tree after intense exercise. Despite the high prevalence of EIPH in horses, the primary aetiology remains unknown. Variants in the genes encoding CD39 and CD39L1 (ENTPD1 and ENTPD2, respectively) were previously reported as potential genetic causes involved in EIPH pathogenesis. However, the role of these variants in haemostatic functions is unknown. RESULTS: To investigate the association between EIPH and missense variants in the ENTPD1 (rs1152296272, rs68621348, and rs68621347) and ENTPD2 genes (rs782872967), 76 Thoroughbred horses diagnosed with EIPH and 56 without clinical signs of EIPH (control group) by trachea-bronchial endoscopy were genotyped. The rs1152296272 and rs68621347 variants were linked, which explained why the same results were found in all horses. Approximately 96% and 95% of the EIPH and control horses, respectively, carried at least one nonreference allele for these variants. In contrast, 100% of the control horses and 96% of the EIPH horses were homozygous for the reference allele for the rs68621348 variant. In the EIPH group, 1.5% of the horses were homozygotes and 24% were heterozygous for the nonreference allele of the rs782872967 variant. In the control group, the nonreference allele of this variant was observed only in heterozygotes (16%). There were no significant differences between groups for any of the variants. CONCLUSIONS: The variants previously described in the genes encoding the CD39 and CD39L1 enzymes were highly present in the studied population. However, no association was found between the occurrence of EIPH and the presence of these variants in Thoroughbred horses in this study.

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Objectives: The aims of the study were to compare the consumption of blood products before and after the implementation of a bleeding management algorithm in patients undergoing liver transplantation and to determine the feasibility of a multicentre, randomized study. Background: Liver transplantation remains the only curative therapy for patients with end-stage liver disease, but it carries a high risk of surgical bleeding. Materials and Methods: Retrospective study of patients treated before (group 1) and after (group 2) implementation of a haemostatic algorithm guided by viscoelastic testing, including use of lyophilized coagulation factor concentrates (prothrombin complex and fibrinogen concentrates). Primary outcome was the number of units of blood products transfused in 24 h after surgery. Secondary outcomes included hospital stay, mortality, and cost. Results: Data from 30 consecutive patients was analysed; 14 in group 1 and 16 in group 2. Baseline data were similar between groups. Median total blood product consumption 24 h after surgery was 33 U (IQR: 11-57) in group 1 and 1.5 (0-23.5) in group 2 (p = 0.028). Significantly fewer units of red blood cells, fresh frozen plasma, and cryoprecipitate were transfused in group 2 versus group 1. There was no significant difference in complications, hospital stay, or in-hospital mortality between groups. The cost of haemostatic therapy was non-significantly lower in group 2 versus group 1 (7,400 vs. 15,500 USD; p = 0.454). Conclusion: The haemostatic management algorithm was associated with a significant reduction in blood product use during 24 h after liver transplantation. This study demonstrated the feasibility and provided a sample size calculation for a larger, randomized study.

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Introducción: La enfermedad trombótica asociada a las hemopatías malignas es subestimada en el paciente grave; en ocasiones queda a la sombra de las hemorragias que presentan estos enfermos. Su diagnóstico y tratamiento constituyen un desafío para lograr el aumento de la sobrevida en las unidades de cuidados críticos Objetivos: Analizar los aspectos generales de las alteraciones trombóticas en pacientes graves con hemopatías malignas. Métodos: Se realizó una investigación bibliográfico-documental acerca del tema. Se consultaron en las bases de datos de SciELO y Pubmed, los artículos publicados en los últimos diez años. Análisis y síntesis de la información: Se describen los aspectos generales vinculados con el mecanismo de la coagulación, sus criterios diagnósticos; así como la evolución del paciente con trombosis que ingresan en las unidades de cuidados intensivos. Conclusiones: El conocimiento de las alteraciones tromboembólicas en los pacientes con hemopatías malignas permite un monitoreo adecuado y la creación de estrategias individuales para mejorar la sobrevida de estos enfermos en la la unidad de cuidados intensivos.

Introduction: Thrombotic disease associated with malignant hemopathies is underestimated in critically ill patients; sometimes it remains in the shadow of the hemorrhages that these patients present. Its diagnosis and treatment constitute a challenge to achieve increased survival in these patients in critical care units Objectives: To analyze the general aspects of thrombotic alterations in seriously ill patients with malignant hemopathies. Methods: A bibliographical-documentary research on the subject was carried out. The SciELO and Pubmed databases of the last ten years were consulted. Analysis and synthesis of the information: General aspects related to the coagulation mechanism, its diagnostic criteria are described; as well as the evolution of the patient with thrombosis who is admitted to the intensive care unit. Conclusions: Knowledge of thromboembolic alterations in patients with malignant hemopathies allows adequate monitoring and the creation of individual strategies to improve the survival of these patients in the ICU.

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INTRODUCTION: The first external quality assessment (EQA) in Thrombosis and Haemostasis was elaborated over 20 years ago, and since then, several national and international EQA institutions have been established. AIM: Display the benefits of EQA programs. METHODS: The spectrum of EQA action was evaluated ranges from improving the performance of the local laboratory to highlighting inadequate diagnostic tests that need to be replaced by new technologies. RESULTS: The first result approach is related to a national management of quality in laboratories. In recent years, Brazil has invested in an EQA program to aid public policy in the laboratory area. During this period, a group of haemostasis laboratory specialists were invited to manage the results and help the Ministry of Health with applying these results as a strategy to improve laboratories. Thus, in collaboration with NEQAS-BC, the University of Campinas - UNICAMP, established a Brazilian EQA program for Blood Coagulation. The second result approach is related to FVIII inhibitor assay performance evaluation, which is another type of EQA program benefit. Despite the assay being considered the gold standard to measure neutralised immunoglobulins for FVIII since 1975, over 40 years ago, the test still has a high coefficient of variation. Results from NEQAS-BC and WFH IEQAS program demonstrate the inter-laboratory variation across the United Kingdom over the last years and among emergent countries. CONCLUSION: The EQA programs have an important educational role in helping countries manage their public policy and in the international inquiry regarding the necessity of new technologies in haemostasis.

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Abstract The haemostatic efficacy of different extract types of Satureja thymbra L., Thymbra spicata L. (Lamiaceae) and Verbascum fruticulosum Post. (Scrophulariaceae) was evaluated in this study via the Prothrombin time (PT) and Activated partial thromboplastin time (aPTT) analysis. Aqueous, methanol and ethanol extracts of the examined plant species leaves were prepared to a final concentration 50 mg/mL. In vitro PT and aPTT assays were conducted on normal platelet poor plasma blood samples by a digital coagulation analyzer. The obtained results revealed anticoagulation activity of all investigated plant species with observed variations among them. The aqueous and ethanol extracts of T. spicata as well as the aqueous extract of S. thymbra prolonged PT values significantly (p < 0.05). While, all V. fruticulosum extract types have had no significant effect on the PT values. The recorded aPTT data showed that all aqueous extracts have had a significant effect on the blood haemostasis as they increased aPTT values in all plant species under study. Out of which, both the ethanol and methanol extracts of T. spicata and methanol extract of S. thymbra showed similar effect. Of great concern, it was clearly noticed that the aqueous and ethanol extract of T. spicata and the aqueous extract of S. thymbra possess the strongest anticoagulation effect as they increased both PT and aPTT values significantly relative to the control (p < 0.05). The variable anticoagulation bioactivity among the studied plant species could be referred to the various solvents degrees of solubility of different phyto-constituents. Thus, the efficacy of the plant species extracts evaluation as anticoagulants or coagulants were related to the plant species and to the solvent of extraction.

Resumo A eficácia hemostática de diferentes tipos de extrato de Satureja thymbra L., Thymbra spicata L. (Lamiaceae) e Verbascum fruticulosum Post. (Scrophulariaceae) foi avaliada neste estudo pelo tempo de protrombina (TP) e tempo de tromboplastina parcial ativada (TTPa). Os extratos aquosos, metanólicos e etanólicos das folhas das espécies de plantas examinadas foram preparados para uma concentração final de 50 mg/mL. Os ensaios de TP e TTPa in vitro foram realizados em amostras normais de sangue, pobre em plaquetas, por um analisador de coagulação digital. Os resultados obtidos revelaram atividade anticoagulante de todas as espécies de plantas investigadas, com variações observadas dentre elas. Os extratos aquosos e etanólicos de T. spicata e o extrato aquoso de S. thymbra prolongaram significativamente os valores de TP (p <0,05). Entretanto, todos os tipos de extratos de V. fruticulosum não tiveram efeito significativo sobre os valores de TP. Os dados registrados do TTPa mostraram que todos os extratos aquosos tiveram um efeito significativo na hemostase do sangue, pois aumentaram os valores de TTPa em todas as espécies de plantas em estudo. Dos quais, ambos os extratos etanólicos e metanólicos de T. spicata e o extrato metanólico de S. thymbra mostraram efeito semelhante. De grande preocupação, notou-se claramente que os extratos aquoso e etanólico de T. spicata e o extrato aquoso de S. thymbra apresentam efeito anticoagulante mais forte, aumentando os valores de TP e TTPa significativamente em relação ao controle (p <0,05). A variável bioatividade anticoagulante dentre as espécies vegetais estudadas pôde ser referida aos vários graus de solventes de solubilidade de diferentes fitoconstituintes. Assim, a eficácia da avaliação de extratos de espécies vegetais como anticoagulantes ou coagulantes foi relacionada às espécies vegetais e ao solvente de extração.

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The haemostatic efficacy of different extract types of Satureja thymbra L., Thymbra spicata L. (Lamiaceae) and Verbascum fruticulosum Post. (Scrophulariaceae) was evaluated in this study via the Prothrombin time (PT) and Activated partial thromboplastin time (aPTT) analysis. Aqueous, methanol and ethanol extracts of the examined plant species leaves were prepared to a final concentration 50 mg/mL. In vitro PT and aPTT assays were conducted on normal platelet poor plasma blood samples by a digital coagulation analyzer. The obtained results revealed anticoagulation activity of all investigated plant species with observed variations among them. The aqueous and ethanol extracts of T. spicata as well as the aqueous extract of S. thymbra prolonged PT values significantly (p < 0.05). While, all V. fruticulosum extract types have had no significant effect on the PT values. The recorded aPTT data showed that all aqueous extracts have had a significant effect on the blood haemostasis as they increased aPTT values in all plant species under study. Out of which, both the ethanol and methanol extracts of T. spicata and methanol extract of S. thymbra showed similar effect. Of great concern, it was clearly noticed that the aqueous and ethanol extract of T. spicata and the aqueous extract of S. thymbra possess the strongest anticoagulation effect as they increased both PT and aPTT values significantly relative to the control (p < 0.05). The variable anticoagulation bioactivity among the studied plant species could be referred to the various solvents degrees of solubility of different phyto-constituents. Thus, the efficacy of the plant species extracts evaluation as anticoagulants or coagulants were related to the plant species and to the solvent of extraction.(AU)

A eficácia hemostática de diferentes tipos de extrato de Satureja thymbra L., Thymbra spicata L. (Lamiaceae) e Verbascum fruticulosum Post. (Scrophulariaceae) foi avaliada neste estudo pelo tempo de protrombina (TP) e tempo de tromboplastina parcial ativada (TTPa). Os extratos aquosos, metanólicos e etanólicos das folhas das espécies de plantas examinadas foram preparados para uma concentração final de 50 mg/mL. Os ensaios de TP e TTPa in vitro foram realizados em amostras normais de sangue, pobre em plaquetas, por um analisador de coagulação digital. Os resultados obtidos revelaram atividade anticoagulante de todas as espécies de plantas investigadas, com variações observadas dentre elas. Os extratos aquosos e etanólicos de T. spicata e o extrato aquoso de S. thymbra prolongaram significativamente os valores de TP (p < 0,05). Entretanto, todos os tipos de extratos de V. fruticulosum não tiveram efeito significativo sobre os valores de TP. Os dados registrados do TTPa mostraram que todos os extratos aquosos tiveram um efeito significativo na hemostase do sangue, pois aumentaram os valores de TTPa em todas as espécies de plantas em estudo. Dos quais, ambos os extratos etanólicos e metanólicos de T. spicata e o extrato metanólico de S. thymbra mostraram efeito semelhante. De grande preocupação, notou-se claramente que os extratos aquoso e etanólico de T. spicata e o extrato aquoso de S. thymbra apresentam efeito anticoagulante mais forte, aumentando os valores de TP e TTPa significativamente em relação ao controle (p < 0,05). A variável bioatividade anticoagulante dentre as espécies vegetais estudadas pôde ser referida aos vários graus de solventes de solubilidade de diferentes fitoconstituintes. Assim, a eficácia da avaliação de extratos de espécies vegetais como anticoagulantes ou coagulantes foi relacionada às espécies vegetais e ao solvente de extração.(AU)

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OBJECTIVE: Malignant wounds due to breast cancer can present with recurrent episodes of bleeding in the tumour tissue. This study will compare the efficacy of a calcium alginate dressing (Biatain, Coloplast A/S, Denmark) and a regenerated oxidised cellulose dressing (Surgicel, Ethicon, LLC, Puerto Rico). PROTOCOL: A total of 24 patients with breast cancer and bleeding, malignant wounds will be enrolled in the randomised, controlled, open study, conducted at a hospital specialising in breast cancer treatment and at another hospital specialising in palliative care. Patients over 18 years old, with bleeding and willing to undergo venipuncture for blood collection will be included. All enrolled patients will be randomised for allocation to an experimental group (regenerated oxidised cellulose dressing) or a control group (calcium alginate dressing). The main intervention will consist of the application of the haemostatic product, assessment of digital pressure and estimation of the time required for haemostasis. OUTCOMES: Key outcome measures will be the percentage of patients with haemostasis within 20 minutes, observation of haemostasis after three, five and 10 minutes, in addition to recurrence of bleeding and the quantity of product used. DISCUSSION: To our knowledge, this is the first study to evaluate the effectiveness of haemostatic products in malignant wounds. This type of wound is poorly explored in the literature and, among its signs and symptoms, bleeding is poorly studied. The completion of this study will provide a more robust rationale for clinical decision-making related to the control of bleeding in malignant breast cancer wounds in the context of evidence-based nursing practices.

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BACKGROUND Calotropis procera latex protein fraction (LP) was previously shown to protect animals from septic shock. Further investigations showed that LP modulate nitric oxide and cytokines levels. OBJECTIVES To evaluate whether the protective effects of LP, against lethal bacterial infection, is observed in its subfractions (LPPII and LPPIII). METHODS Subfractions (5 and 10 mg/kg) were tested by i.p. administration, 24 h before challenging with lethal injection (i.p.) of Salmonella Typhimurium. LPPIII (5 mg/kg) which showed higher survival rate was assayed to evaluate bacterial clearance, histopathology, leukocyte recruitment, plasma coagulation time, cytokines and NO levels. FINDINGS LPPIII protected 70% of animals of death. The animals given LPPIII exhibited reduced bacterial load in blood and peritoneal fluid after 24 h compared to the control. LPPIII promoted macrophage infiltration in spleen and liver. LPPIII restored the coagulation time of infected animals, increased IL-10 and reduced NO in blood. MAIN CONCLUSIONS LPPIII recruited macrophages to the target organs of bacterial infection. This addressed inflammatory stimulus seems to reduce bacterial colonisation in spleen and liver, down regulate bacterial spread and contribute to avoid septic shock.

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Objective: to describe the haemostatic characteristics of the venom as well as the potency appraisal of the polyvalent antiophidic serum against haemotoxicity from Porthidium lansbergii hutmani experimental envenomation. Methods: Evaluation was performed of the venom's lethality, haemorrhagic activity, effects on coagulation and platelet aggregation, proteolytic activity, and neutralization by the commercial antivenom available in the country. Results: Several components with haemostatic activities were found in Porthidium l. hutmanni venom when a study of fibrinogenolytic, haemorrhagic and proteolytic activities was conducted of a pool of P.l.h venom. Porthidium l. hutmanni venom lacked the coagulant and defibrinating activities that are characteristic of bothropic venoms. Porthidium l. hutmanni venom showed very high haemorrhagic and anticoagulant activities. These findings could be related to the presence of multiple metalloproteases, which was evidenced in this study, and also the possible presence of phospholipases or other anticoagulant activity proteins that were not defined here. They inhibited platelet aggregation, suggesting that the venom had some proteins with marked effects on haemostasis. The commercial antivenom proved to be of little effectiveness in neutralizing the crude venom haemorrhagic activity. Conclusions: These toxins cause many physiopathological alterations in bitten patients, creating a clinical picture characterized by oedema, local and systemic haemorrhages, and even necrosis, comparable to that seen in bothropic envenomation. Porthidium l. hutmanni venom has no in vitro procoagulant activity, typical of bothropic venoms, suggesting there are variances in its protein conformation. Porthidium l. hutmanni venom is used for horse immunization. However, in order to preserve the patient's life, it is necessary to improve the immunization process to produce antivenom containing high avidity and specificity antibodies against the major toxins present in this venom. Porthidium l. hutmanni venom has demonstrated being a venom with high lethal, haemorrhagic, proteolytic and procoagulant activities, whose description will have enormous utility among clinicians who deal with these accidents in its geographical distribution areas(AU)

Objetivo: Describir las características hemostáticas del veneno y evaluar la potencia del suero polivalente antiofídico contra la hemotoxicidad provocada por el envenenamiento experimental por Porthidium lansbergii hutmanni. Métodos: Se realizó una evaluación de la letalidad, actividad hemorrágica, efectos en la coagulación y agregación plaquetaria, actividad proteolítica y neutralización por el antiveneno disponible comercialmente en el país. Resultados: Se encontraron varios componentes con actividad hemostática en el veneno de Porthidium l. hutmanni al realizarse un estudio de la actividad fibrinogenolítica, hemorrágica y proteolítica de una muestra de veneno de Porthidium l. hutmanni. El veneno de Porthidium l. hutmanni no mostró la actividad coagulante o defibrinante característica de los venenos botrópicos. El veneno de Porthidium l. hutmanni mostró una elevada actividad hemorrágica y anticoagulante. Estos resultados podrían estar relacionados con la presencia de múltiples metaloproteasas, la que quedó demostrado en el estudio, y también a la posible presencia de fosfolipasas u otras proteínas de actividad anticoagulante que no se definen en el mismo. La inhibición de la agregación plaquetaria sugiere que el veneno contiene algunas proteínas con un marcado efecto sobre la hemostasis. El antiveneno comercial mostró poca efectividad en la neutralización de la actividad hemorrágica del veneno crudo. Conclusiones: Estas toxinas provocan muchas alteraciones fisiopatológicas en las víctimas de mordeduras, creando un cuadro clínico caracterizado por edema, hemorragias locales y sistémicas e incluso necrosis comparable con la que ocurre en el envenenamiento botrópico. El veneno de Porthidium l. hutmanni no tiene la actividad procoagulante in vitro típica de los venenos botrópicos, lo que apunta a variaciones en su conformación proteica. El veneno de Porthidium l. hutmanni de utiliza en la inmunización de los caballos. Sin embargo, para preservar la vida del paciente, es necesario mejorar el proceso de inmunización con vistas a producir un antiveneno que contenga anticuerpos de elevada avidez y especificidad contra las principales toxinas presentes en el veneno. El veneno de Porthidium l. hutmanni ha mostrado ser un veneno de elevada actividad letal, hemorrágica, proteolítica y procoagulante, cuya descripción tendrá una enorme utilidad para los médicos que atienden esos accidentes en sus áreas de distribución geográfica(AU)

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Natural enzyme inhibitors have been widely described in literature because of its pharmacological and cosmetic applications. Fungi found in caves represent a promising source of bioactive substances that are still little explored scientifically. Thus, the present work evaluated the presence of enzymatic modulators in a filtrate obtained from the cultivation of the cave fungus Lecanicillium aphanocladii (Family: Cordycipitaceae). Snake venoms from Bothrops alternatus and Bothrops atrox were used as an enzymatic source for the induction of the phospholipase, proteolytic, thrombolytic, cytotoxic and coagulant activities. Compounds present in the fungal filtrate inhibited 50, 23·8, 26·6, 50·9 and 52·5% of the proteolytic, phospholipase, haemolytic, thrombolytic and coagulant activities respectively. The filtrate was not cytotoxic on erythrocytes, but induced partial dissolution of thrombi. Fungal enzyme inhibitors that have low or no toxicity and can be obtained on a large scale and at low cost are relevant in the medical-scientific context. Therefore, the inhibition of phospholipases A2 and proteases observed in the present work highlights the potential of fungal metabolites for the development of drugs that can be used in the treatment of haemostasis and inflammation-related disorders. SIGNIFICANCE AND IMPACT OF THE STUDY: In this study, secondary metabolites synthesized by Lecanicillium aphanocladii, a fungus isolated from caves, demonstrated modulating action on proteases and phospholipases A2 present in snake venoms of the Bothrops genus, widely used as tools for the study of pathophysiology processes related to haemostasis and inflammation. The results suggest the possibility of future applications for these metabolites in the development of pharmaceuticals of medical-scientific interest.

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