Comparing modified Lee and White method against 20-minute whole blood clotting test as bedside coagulation screening test in snake envenomation victims

Abstract Background: Twenty-minute whole blood clotting test (20WBCT) and Modified Lee and White (MLW) method are the most routinely employed bedside tests for detecting coagulopathic snake envenomation. Our study compared the diagnostic utility of MLW and 20WBCT for snakebite victims at a tertiary care hospital in Central Kerala, South India. Methods: This single-center study recruited 267 patients admitted with snake bites. 20WBCT and MLW were performed simultaneously at admission along with the measurement of Prothrombin Time (PT). The diagnostic utility of 20WBCT and MLW was determined by comparing the sensitivity (Sn), specificity (Sp), positive and negative predictive values, likelihood ratios, and accuracy at admission with an INR value > 1.4. Results: Out of 267 patients, 20 (7.5%) patients had VICC. Amongst those who had venom-induced consumption coagulopathy (VICC), MLW was prolonged for 17 patients, (Sn 85% 95% confidence interval [CI]: 61.1-96.0) whereas 20WBCT was abnormal for 11 patients (Sn 55%, 95% CI: 32.04-76.17). MLW and 20WBCT were falsely positive for the same patient (Sp 99.6%, 95% CI: 97.4-99.9%). Conclusion: MLW is more sensitive than 20WBCT to detect coagulopathy at the bedside amongst snakebite victims. However, further studies are necessary for standardizing bedside coagulation tests in snakebite cases.

El papel de la enfermedad de von Willebrand en la hemorragia uterina anormal / Von Willebrand Disease in Abnormal Uterine Bleeding

RESUMEN La hemorragia uterina anormal es un término empleado para las alteraciones en la regularidad, duración y/o volumen de sangrado menstrual y es considerada una causa común de consulta médica y en ocasiones supone un reto diagnóstico para el médico tratante. Dentro del abordaje de la etiología de dicha patología, las coagulopatías afectan alrededor del 13 por ciento de las mujeres, y la más común es la enfermedad de von Willebrand. El objetivo de este trabajo fue realizar una revisión de la literatura científica actual sobre el papel que cumple la enfermedad de von Willebrand en la hemorragia uterina anormal. Esta es una patología hereditaria derivada de una deficiencia del factor von Willebrand encargado de la adhesión plaquetaria. La prevalencia de esta enfermedad puede ser baja, sin embargo, cuando se estudia la población de mujeres con menorragia, la frecuencia puede ir de 5 a 20 por ciento. Se han descrito diferentes problemas ginecológicos asociados a la enfermedad de von Willebrand, tales como menorragia, dismenorrea y una importante deficiencia de hierro asociada a esta, además de una mayor incidencia de quistes ováricos, endometriosis, hiperplasia endometrial y pólipos endometriales. La literatura actual sugiere que se realice tamizaje a aquellas mujeres con cuadro clínico sugestivo. Con respecto al tratamiento la literatura reporta el uso de ácido tranexámico y anticonceptivos orales, pero el que mayor utilidad ha demostrado es la desmopresina(AU)

ABSTRACT Abnormal uterine bleeding is a term used for alterations in the regularity, duration and / or volume of menstrual bleeding and it is considered a common cause of medical consultation; sometimes it is a diagnostic challenge for the treating physician. Within the aetiology approach of said pathology, coagulopathies affect around 13 percent of women, and the most common is von Willebrand disease. The objective is to review the current scientific literature on the influence of von Willebrand disease in abnormal uterine bleeding. This is an inherited pathology derived from a deficiency of the von Willebrand factor responsible for platelet adhesion. The prevalence of this disease may be low, however, when studying the population of women with menorrhagia, the frequency can range from 5 to 20 percent. Different gynecological problems associated with von Willebrand disease have been described, such as menorrhagia, dysmenorrhea and a significant iron deficiency associated with it, in addition to a higher incidence of ovarian cysts, endometriosis, endometrial hyperplasia and endometrial polyps. The current literature suggests that those women with suggestive clinical symptoms should be screened. Regarding treatment, the literature reports the use of tranexamic acid and oral contraceptives, nonetheless desmopressin has proven to be most useful(AU)

Hemofilia: abordaje diagnóstico y terapéutico. Revisión bibliográfica / Hemophilia: Diagnostic and Therapeutic Approach. A Literature review / Hemofilia: aproximação diagnóstica e terapêutica. Revisão bibliográfica

Resumen Antecedentes: La hemofilia es una enfermedad de origen genético, recesiva y ligada al cromosoma X; su principal manifestación clínica es la hemorragia, cuyo grado depende del nivel del factor VIII o IX que se halla en el plasma, usualmente secundaria a traumas en sitios de localización profunda, como articulaciones, músculos y sistema nervioso central. Objetivo: El objetivo es presentar una revisión bibliográfica clara y práctica de la hemofilia, donde se abordan aspectos generales de la fisiopatología, el diagnóstico y el manejo, al igual que las nuevas alternativas terapéuticas en desarrollo para su tratamiento. Metodología: Se llevó a cabo una búsqueda en las bases de datos PubMed, ScienceDirect y Scielo, a partir de las palabras clave en español e inglés. Se seleccionaron 32 artículos que fueron la base para la construcción de este manuscrito. Resultados: Se construyó una revisión bibliográfica que incluye conceptos básicos y prácticos para el enfoque y el abordaje de la hemofilia. Conclusión: La hemofilia es una entidad potencialmente mortal, que afecta la calidad de vida de los pacientes y de aquellos que lo rodean; es un reto hacer un enfoque adecuado para el diagnóstico y el tratamiento.

Abstract Background: Hemophilia is a genetic, recessive disorder linked to the X chromosome. Its main clinical symptom is he morrhage, whose degree depends on the level of factor VIII or IX found in plasma, usually following traumas in deep sites, such as joints, muscles and central nervous system. Objecti ve: The objective is to present a clear and practical literature review of hemophilia, with general aspects of its pathophysio logy, diagnosis and treatment, as well as the new therapeutic alternatives being developed for its treatment. Methodology: the PubMed, ScienceDirect and Scielo databases were chec ked, using keywords in Spanish and English. We selected 32 articles, which were the basis for constructing this text. Re sults: a bibliographic review including basic and practical concepts for the approach and treatment of hemophilia was constructed. Conclusion: hemophilia is a potentially deadly disorder, affecting the quality of life of patients and those around them as finding a proper approach for its diagnosis and treatment is challenging.

Resumo Antecedentes: a hemofilia é uma doença de origem genética, re cessiva e ligada ao cromossoma X. A sua principal manifestação clínica é a hemorragia, cujo grau depende do nível do fator VIII ou IX achado no plasma, usualmente secundária a traumas em lugares profundos, como articulações, músculos e o sistema ner voso central. Objetivo: apresentar uma revisão bibliográfica clara e prática da hemofilia, com aspetos gerais da fisiopatologia, do diagnóstico e da gestão, também de novas alternativas de terapia em desenvolvimento para o tratamento. Metodologia: pesquisou-se nas bases de dados PubMed, ScienceDirect e Scielo, com pala vras chaves no espanhol e no inglês. Selecionaram-se 32 artigos, que foram a base para construir este texto. Resultados: construiu-se uma revisão bibliográfica incluindo conceitos básicos e prá ticos para o enfoque e o tratamento da hemofilia. Conclusão: a hemofilia é uma entidade potencialmente mortal, afetando a qua lidade de vida dos pacientes e dos que estão ao seu redor, é um desafio fazer uma aproximação apropriada para o diagnóstico e para o tratamento.

Delayed double reading of whole blood clotting test (WBCT) results at 20 and 30 minutes enhances diagnosis and treatment of viper evenomation

The whole blood clotting test (WBCT) is a simple test of coagulation that is often used in the assessment, diagnosis, and therapeutic monitoring of snakebite patients in sub-Saharan Africa. WBCT requires only a clean glass tube and several milliliters of venous blood and is ideal for use in poorly equipped health centers throughout the rural areas where 95% of snakebites occur. However, questions surrounding the accuracy and reliability of the test remain unanswered due to variations in testing conditions and a lack of comparative research with which to validate them. This is the first study to evaluate WBCT results at both 20-min (WBCT20) and 30-min (WBCT30) reading times in the same group of snakebite patients. Methods In order to define the best reading time, the authors compared the results of serial WBCT evaluation at both 20 and 30 min after collection in 23 patients treated for snake envenomation in Bembèrèkè, northern Benin. Results WBCT results were identical at both reading times in patients without coagulopathy or when coagulation was restored permanently following a single dose of antivenom. Out of 17 patients with coagulopathy, 14 showed discrepancies between WBCT20 and WBCT30 results in at least one pair of serial evaluations. These could be completely contradictory results (e.g. normal clot at WBCT20 and no clot at WBCT30) or a marked difference in the quality of the clot (e.g. no clotting activity at WBCT20 and an unstable partial clot at WBCT30). WBCT discrepancies were encountered most frequently in three situations: initial normalization of hemostasis following antivenom therapy, detection of a secondary resumption of coagulopathy, or final restoration of hemostasis after a secondary resumption had occurred. Conclusions This study suggests that the WBCT is robust and that a sequential reading should improve the diagnosis and monitoring of venom-induced coagulopathies. It also indicates the possibility of discrepancies in the sensitivity of WBCT20 and WBCT30 for detecting the resolution or reoccurrence of coagulopathy and identifies how these findings, if confirmed, may be used to increase the efficacy and efficiency of antivenom treatment in the field.(AU)

Effects of four extracts of Crataegi Folium on blood stasis syndrome related indexes based on principal component analysis / 中草药

Objective To observe the effects of four kinds of extracts of Crataegi Folium on hemorheology, erythrocytes, platelets, coagulation time, and other indicators of 27 indicators in blood stasis rats from the aspects of whole and local double aspects by using the combination method of principal component and one-way ANOVA, and analysis of indicators contained in the information, in order to provide new analytical ideas for the mechanism of Crataegi Folium extract on activating blood and resolving stasis. Methods Forty-eight wistar rats were randomly divided into blank group, model group, and drug group. Blood stasis model was prepared by using adrenaline in ice water bath. Both blank group and model group were given normal saline, and drug group was given hawthorn leaves extract aqueous solution. Blood samples were collected and 27 indicators such as hemorheology, erythrocyte, platelet, and prothrombin time were selected. The principal component analysis (PCA) was used to decompose all the index data by dimensionless treatment and dimensionality reduction, and then the principal components were generated. The new variables (ie, principal component scores) were subsequently analyzed by one-way ANOVA. The results combined with the contribution rate of principal components and the correlation coefficient of each index was analyzed. Finally, the index groups and individual indexes in each group were analyzed synthetically, and the regularity conclusions were drawn. Results Eight principal components were extracted by principal component analysis, and the contribution rate was 77.992%. The results of theprincipal component analysis combined with the ANOVA showed that nine main components in the first principal component could replace the original 27 indicators to reflect the effect of Crataegi Folium extract on blood circulation and blood stasis, namely FIB, total number of red blood cells, total platelet count platelet volume, platelet count, blood viscosity at low shear rate, blood viscosity at middle shear rate, blood viscosity at high shear rate, and plasma viscosity. The results showed that the effects of the four extracts of Crataegi Folium on the index components were different. Conclusion The effect of activating blood and resolving stasis is mainly by reducing blood viscosity and plasma viscosity, promoting hemoglobin degradation and increasing the number of platelets, shortening the blood clotting and so on to achieve.

Delayed double reading of whole blood clotting test (WBCT) results at 20 and 30 minutes enhances diagnosis and treatment of viper evenomation

The whole blood clotting test (WBCT) is a simple test of coagulation that is often used in the assessment, diagnosis, and therapeutic monitoring of snakebite patients in sub-Saharan Africa. WBCT requires only a clean glass tube and several milliliters of venous blood and is ideal for use in poorly equipped health centers throughout the rural areas where 95% of snakebites occur. However, questions surrounding the accuracy and reliability of the test remain unanswered due to variations in testing conditions and a lack of comparative research with which to validate them. This is the first study to evaluate WBCT results at both 20-min (WBCT20) and 30-min (WBCT30) reading times in the same group of snakebite patients. Methods In order to define the best reading time, the authors compared the results of serial WBCT evaluation at both 20 and 30 min after collection in 23 patients treated for snake envenomation in Bembèrèkè, northern Benin. Results WBCT results were identical at both reading times in patients without coagulopathy or when coagulation was restored permanently following a single dose of antivenom. Out of 17 patients with coagulopathy, 14 showed discrepancies between WBCT20 and WBCT30 results in at least one pair of serial evaluations. These could be completely contradictory results (e.g. normal clot at WBCT20 and no clot at WBCT30) or a marked difference in the quality of the clot (e.g. no clotting activity at WBCT20 and an unstable partial clot at WBCT30)...

Accidente ofídico en pacientes del Hospital Básico de Jipijapa (Manabí-Ecuador) / Snake accident in patients of the Basic Hospital of Jipijapa (Manabí-Ecuador)

Introducción: el accidente ofídico es el cuadro clínico producido por la mordedura de una serpiente venenosa. En el Ecuador se describen 41 especies de serpientes venenosas. Objetivo: caracterizar clínica y epidemiológicamente el accidente ofídico en pacientes del Hospital Básico de Jipijapa (Manabí-Ecuador). Método: se realizó un estudio observacional retrospectivo durante enero 2008 - abril 2012, a partir de las historias clínicas de los pacientes hospitalizados con diagnóstico de accidente ofídico o mordedura de serpiente en el Servicio de Medicina Interna del Hospital Básico de Jipijapa (Manabí-Ecuador). Resultados: del total de 78 pacientes, 61 (78,2%) recibieron suero antiofídico (SAO) y 17 pacientes (21,8%) no recibieron. No se encontró relación entre los días de hospitalización y el hecho de haber o no recibido SAO (p= 0,8), ni tampoco entre la prueba de coagulación positiva o negativa con el tiempo de hospitalización (p= 0,7). Se encontró una correlación baja entre la variable prueba de coagulación y el grado de mordedura, siendo estadísticamente significativo (p= 0,04). Se observó incremento de mordeduras en ciertos meses, en relación con los hábitos migratorios y reproductivos de las serpientes. Conclusiones: existió dificultad en la aplicación de protocolos de atención para accidente ofídico y si bien esto no repercutió en la morbilidad y mortalidad, puede afectar en la optimización de los recursos disponibles.

Introduction: ophidian accident is the clinical picture caused by the bite of a venomous snake. Forty one species of poisonous snakes are described in Ecuador Objective: to describe clinically and epidemiologically the ophidian accident in patients of the Basic Hospital of Jipijapa (Manabí-Ecuador). Methods: a retrospective observational, descriptive study, based on medical records of hospitalized patients with a diagnosis of snake bite in Internal Medicine Basic Hospital, Jipijapa (Manabí-Ecuador) during January 2008 - April 2012 was done. Results: of the total 78 patients, 61 (78.2%) received antivenin and 17 patients (21.8%) did not receive. There was no relationship between days of hospitalization and the fact of having or not received antivenin, p=Ns (0.8) nor between test positive or negative coagulation time found hospitalization p=Ns (0.7). A low correlation was found between the coagulation test variable and the degree of bite, being statistically significant (p = 0.04). An increase of bites in certain months, in relation to the migratory and reproductive habits of the snakes was observed. Conclusions: there was difficulty in applying protocols of care for ophidian accidents and although this did not affect morbidity and mortality, it can affect the optimization of available resources.

Comprometimento hemostático em pacientes com icterícia obstrutiva / Haemostatic impairment in patients with obstructive jaundice

A doença hepática é associada à alterações hemostáticas devido à diminuição na síntese dos fatores de coagulação. Neste sentido, objetivou-se avaliar o comprometimento da hemostasia em pacientes com icterícia obstrutiva. Os pacientes foram analisados por gênero, idade, etiologia da doença, tempo de protrombina (TP) e tempo de tromboplastina parcial (TTP) antes e após a cirurgia. A icterícia obstrutiva ocorreu com maior frequência entre as mulheres (60,0%), sendo a coledocolitíase a causa mais comum da doença (80,0%). Os valores pré-cirúrgicos do TTP estavam dentro da faixa normal. No entanto, todos os pacientes apresentaram um TP prolongado na avaliação pré-operatória. Uma redução significativa (p<0,001) foi verificada na análise pós-operatória do TP, atingindo a faixa normal em 66,7% dos pacientes estudados. Verificou-se uma correlação positiva (r = 0,813 p<0,001) entre os valores pré-operatórios do TP e a idade dos pacientes. Além disso, os valores pré-operatórios e pós-operatórios do TP foram significativamente mais elevados (p<0,05) entre pacientes com icterícia obstrutiva de etiologia tumoral. Os resultados deste estudo demonstraram uma forte correlação entre a idade dos pacientes e a atividade dos fatores de coagulação vitamina K dependentes, além de uma diminuição significativa na atividade destes fatores em pacientes com icterícia obstrutiva de etiologia tumoral.

Liver disease is associated with haemostatic abnormalities due to the decreased synthesis of the clotting factors. Therefore, this study aimed to evaluate the impairment of haemostasis in patients with obstructive jaundice. Patients were analyzed by gender, age, cause of jaundice, prothrombin time (PT) and partial thromboplastin time (PTT) before and after surgery. Obstructive jaundice occurred more commonly amongst the females (60.0%), and choledocholithiasis was the most frequent cause of the disease (80.0%). Presurgical PTT values were within the normal range, whereas all the patients showed increased PT in the preoperative evaluation. A significant decrease (p<0.001) was found in the postoperative PT measurement, reaching the normal range in 66.7% of the studied patients. A positive correlation was found between the preoperative PT values and the age of the patients (r = 0.813 p<0.001). In addition, preoperative and postoperative PT values were significantly higher (p<0.05) among the patients with obstructive jaundice of tumoral etiology. Results from this study have shown that activity of vitamin K-dependent clotting factors in obstructive jaundice was strongly correlated with subject's age and was significantly decreased in patients with tumoral etiology.

Clinical and biological analysis in graftless maxillary sinus lift

Maxillary sinus lift for dental implant installation is a well-known and versatile technique; new techniques are presented based on the physiology of intrasinus bone repair. The aim of this review was to determine the status of graftless maxillary sinus lift and analyze its foundations and results. A search was conducted of the literature between 1995 and 2015 in the Medline, ScienceDirect, and SciELO databases using the keywords “maxillary sinus lift,”“blood clot,”“graftless maxillary sinus augmentation,” and “dental implant placement.” Ten articles were selected for our analysis of this technique and its results. Despite the limited information, cases that were followed for at least six months and up to four years had a 90% success rate. Published techniques included a lateral window, elevation of the sinus membrane, drilling and dental implant installation, descent of the membrane with variations in the installation of the lateral wall access and suturing. The physiology behind this new bone formation response and the results of the present research were also discussed. We concluded that this is a promising and viable technique under certain inclusion criteria.

Na2EDTA anticoagulant impaired blood samples from the teleost piaractus mesopotamicus / Anticoagulante Na2EDTA danifica amostras de sangue do teleósteo piaractus mesopotamicus

The present study aimed to evaluate the effects of Na heparin and Na2EDTA on blood of Piaractus mesopotamicus (360.7±42.4g, 26.4±1.0cm). Twenty fishes were sampled in two experiment trials, ten for erythrocyte fragility analysis and ten for hematologic and plasma biochemical study. The blood collected by venous-caudal puncture was fractioned and stored in anticoagulants solution: Na2EDTA 10%, Na2EDTA 3%, Na heparin 5000 IU and Na heparin 100 IU. Plasmatic levels of calcium presented in the Na2EDTA stored samples were about 80% lower than both heparin groups. Blood samples of P. mesopotamicus stored with Na2EDTA demonstrated increase in the hematocrit and MCV, and decrease in MCHC. The dose-response effect was observed in this study. The results are reinforced by the higher levels of plasmatic protein and hemolysis presented in the Na2EDTA 10% stored blood, confirming the deleterious effect of this anticoagulant treatment on the quality of blood samples. Na2EDTA is not indicated to store P. mesopotamicus blood samples, but sodium heparin at 100 IU is the most recommended anticoagulant, since this treatment presented the lower rate of alterations in the stored blood.

O presente estudo teve por objetivo avaliar os efeitos da heparina sódica e Na2EDTA no sangue de Piaractus mesopotamicus (360,7±42,4g, 26,4±1,0cm). Foram amostrados vinte peixes em dois ensaios experimentais, sendo dez peixes utilizados para análise da fragilidade dos eritrócitos e dez peixes para análise dos parâmetros hematológicos e estudo bioquímico do plasma. O sangue coletado por punção veno-caudal foi aliquotado e armazenado em diferentes soluções anticoagulantes: Na2EDTA 10%, Na2EDTA 3%, heparina sódica 5.000 UI e heparina sódica 100 UI. Níveis plasmáticos de cálcio apresentados nas amostras armazenados em Na2EDTA diminuíram cerca de 80% em relação aos dois grupos armazenados com heparina. Amostras de sangue de pacus armazenados com Na2EDTA demonstraram aumento do hematócrito e VCM, e diminuição na CHCM. O efeito dose-resposta foi observado neste estudo. Estes resultados são reforçados pelos níveis mais elevados de proteína plasmática e hemólise apresentado no sangue armazenado com Na2EDTA 10%, o que confirma o efeito deletério desse tratamento anticoagulante na qualidade de amostras de sangue. Na2EDTA não é indicada para armazenar amostras de sangue de P. mesopotamicus, e heparina sódica a 100 UI é o anticoagulante mais recomendado, uma vez que este tratamento apresentou a menor taxa de alterações no sangue armazenado.

Effect and Mechanism of Extracts of Nidus Collocaliae on Resisting Avian Influenza A H5N1 Virus / 广州中医药大学学报

Objective To observe the inhibitory effect of the extracts of Nidus Collocaliae on avian influenza A H5N1 virus in vitro. Methods Nidus Collocaliae water extract, artificial gastric juice digestion products of Nidus Collocaliae water extract, and artifitial intestinal juice digestion products of Nidus Collocaliae water extract were prepared for the experimental study. 293T cells transfection in vitro was carried out. The effects of 3 kinds of Nidus Collocaliae extracts on H5N1 pseudovirus and VSV-G pseudovirus were determined by luciferase detection kit. Blood clotting response to erythrocyte hemagglutinin subtypes H5, H7, H9 antigens and the inhibitory effects of 3 kinds of Nidus Collocaliae extracts were observed. The effects of 3 kinds of Nidus Collocaliae extracts on neuraminidase activity were determined by neuraminidase inhibitor screen kit. Results The 3 kinds of Nidus Collocaliae extracts had inhibitory effects on H5N1 avian influenza pseudovirus, the effects being enhanced with the increase of the concentrations of Nidus Collocaliae extracts. Of the 3 extracts, artificial intestinal digestion products had the strongest inhibitory effect, while Nidus Collocaliae water extract had the weakest effect. However, Nidus Collocaliae extracts had no obvious effect on VSV-G pseudovirus. The concentration of H5, H7 antigen for positive blood clotting response was 1 ∶ 128, and that of H9 antigen was 1 ∶ 256. The 3 kinds of Nidus Collocaliae extracts at certain concentrations could inhibit blood clotting response to H5, H7, H9 antigen, but had no obvious effect on neuraminidase. Conclusion The anti-H5N1 virus effect of Nidus Collocaliae extracts has been achieved probably through resisting hemagglutinin.

An Unusual Presentation of Vasculotoxic Snake Bite.

In India, viper bites are more common than any other poisonous snake bites. In these patients, the cardinal features are local pain, swelling and mild coagulation abnormalities; but gangrenous changes involving bilateral upper and lower limbs are uncommon. We are reporting a case of a 62-year-old man bitten by a Russell viper on the right thumb. He experienced local pain, swelling and later developed disseminated intravascular coagulopathy, pre-renal azotemia, sepsis, acute respiratory distress syndrome (ARDS) and peripheral gangrenous changes involving bilateral upper and lower limbs. He succumbed to his ailment 5 months after the bite. There have been very few cases of gangrenous changes involving distal part of extremities associated with snake bite.

Isolated biomolecules of pharmacological interest in hemostasis from Cerastes cerastes venom

Biomolecules from Cerastes cerastes venom have been purified and characterized. Two phospholipases isolated from Cerastes cerastes venom share 51% of homology. CC2-PLA2 exhibits antiplatelet activity that blocks coagulation. CCSV-MPase, a non-hemorrhagic Zn2+-metalloproteinase, significantly reduced the plasmatic fibrinogen level and hydrolyzes only its Bß chain. Serine proteinases such as RP34, afaâcytin and CC3-SPase hydrolyze the fibrinogen and are respectively α, αß and αß fibrinogenases. In deficient human plasma, afaâcytin replaces the missing factors VIII and IX, and activates purified human factor X into factor Xa. It releases serotonin from platelets and directly aggregates human (but not rabbit) blood platelets. RP34 proteinase also had no effect on both human and rabbit blood platelet aggregation. CC3-SPase revealed a pro-coagulant activity. However, the insolubility of the obtained clot indicates that CC3-SPase does not activate factor XIII. In addition, CC3-SPase clotting activity was carried out with human plasmas from volunteer patients deficient in clotting factors. Results showed that CC3-SPase shortens clotting time of plasma deficient in factors II and VII but with weaker clotting than normal plasma. The clotting time of plasma deficient in factor II is similar to that obtained with normal plasma; suggesting that CC3-SPase is able to replace both factors IIa and VII in the coagulation cascade and thus could be involved in the blood clotting process via an extrinsic pathway. These results imply that CC3-SPase and afaâcytin could repair hemostatic abnormalities and may replace some factors missing in pathological deficiency. Afaâcytin also exhibits α fibrinase property similar to a plasmin-like proteinase. Despite its thrombin-like characteristics, afaâcytin is not inhibited by plasmatic thrombin inhibitors. The procoagulant properties of afaâcytin might have potential clinical applications.(AU)

Isolated biomolecules of pharmacological interest in hemostasis from Cerastes cerastes venom

Biomolecules from Cerastes cerastes venom have been purified and characterized. Two phospholipases isolated from Cerastes cerastes venom share 51% of homology. CC2-PLA2 exhibits antiplatelet activity that blocks coagulation. CCSV-MPase, a non-hemorrhagic Zn2+-metalloproteinase, significantly reduced the plasmatic fibrinogen level and hydrolyzes only its Bβ chain. Serine proteinases such as RP34, afaâcytin and CC3-SPase hydrolyze the fibrinogen and are respectively α, αβ and αβ fibrinogenases. In deficient human plasma, afaâcytin replaces the missing factors VIII and IX, and activates purified human factor X into factor Xa. It releases serotonin from platelets and directly aggregates human (but not rabbit) blood platelets. RP34 proteinase also had no effect on both human and rabbit blood platelet aggregation. CC3-SPase revealed a pro-coagulant activity. However, the insolubility of the obtained clot indicates that CC3-SPase does not activate factor XIII. In addition, CC3-SPase clotting activity was carried out with human plasmas from volunteer patients deficient in clotting factors. Results showed that CC3-SPase shortens clotting time of plasma deficient in factors II and VII but with weaker clotting than normal plasma. The clotting time of plasma deficient in factor II is similar to that obtained with normal plasma; suggesting that CC3-SPase is able to replace both factors IIa and VII in the coagulation cascade and thus could be involved in the blood clotting process via an extrinsic pathway. These results imply that CC3-SPase and afaâcytin could repair hemostatic abnormalities and may replace some factors missing in pathological deficiency. Afaâcytin also exhibits α fibrinase property similar to a plasmin-like proteinase. Despite its thrombin-like characteristics, afaâcytin is not inhibited by plasmatic thrombin inhibitors. The procoagulant properties of afaâcytin might have potential clinical applications.

Heparina e K3EDTA como anticoagulantes para tambaqui (Colossoma macropomum Cuvier, 1816) / Heparin and K3EDTA as anticoagulants for tambaqui (Colossoma macropomum Cuvier, 1816)

Este estudo avaliou a eficácia da heparina sódica e do EDTA tripotássico como anticoagulantes e seus efeitos sobre os parâmetros hematológicos de tambaqui (Colossoma macropomum). Foram utilizados dez indivíduos de tambaqui com médias de 384,9 ± 85,71 g de peso e 27,90 ± 2,10 cm de comprimento total para avaliação da heparina 5.000 UI e 100 UI, bem como do K3EDTA 10 por cento. Foram analisados a inibição da coagulação por 10 h, eritrograma e teste de fragilidade osmótica dos eritrócitos. Os resultados foram submetidos à análise de variância e as médias comparadas pelo teste de Tukey (P < 0,05). Heparina 5000 UI, heparina 100 UI e K3EDTA 10 por cento foram eficazes na prevenção da coagulação por mais de 10 h, no entanto o EDTA tripotássico causou hemólise desde os primeiros momentos. No eritrograma não foi observada diferença (P > 0,05) na contagem de eritrócitos, hematócrito, taxa de hemoglobina e CHCM, no entanto, houve aumento do VCM (P < 0,05) nas amostras acondicionados com K3EDTA 10 por cento. Este anticoagulante causou incremento (P < 0,01) na fragilidade osmótica dos eritrócitos quando comparado com a heparina pura, heparina diluída, e grupo controle. A utilização da heparina como anticoagulante é mais apropriada para tambaqui (Colossoma macropomum), visto que foi eficiente na prevenção da coagulação por mais de 10 h, sem ocasionar hemólise, ou alterações nos parâmetros hematológicos e na fragilidade osmótica dos eritrócitos.

The efficacy of sodium heparin and tripotassium EDTA as anticoagulant and their effect on the hematological parameters of tambaqui (Colossoma macropomum) were evaluated in this study. Ten fish weighing 384.9 ± 85.71 g and measuring 27.90 ± 2.10 cm were used for heparin 5.000 IU, heparin 100 IU and K3EDTA 10 percent evaluation. Clotting inhibition after 10 h, erythrogram and osmotic fragility of erythrocytes were observed. The results were submitted to variance analysis and means compared by Tukey test (P < 0.05). Heparin 5.000 IU, heparin 100 IU and K3EDTA 10 percent were effective in preventing coagulation for more than 10 h. However, tripotassium EDTA caused hemolysis since first moments. In erythrogram there was no difference (P > 0.05) in erythrocyte count, hematocrit, hemoglobin and MCHC. On the other hand, an increase in MCV (P < 0.05) in samples kept with K3EDTA10 percent was observed. This anticoagulant provoked a significant increase (P < 0.01) in the osmotic fragility of erythrocytes when compared to pure heparin, diluted heparin and the control group. Heparin as an anticoagulant is more appropriate for tambaqui since it was effective in preventing coagulation for more than 10 h, without causing hemolysis, changes on hematological parameters or osmotic fragility of erythrocytes.

Hemostasia normal y coagulación intravascular diseminada en obstetricia / Normal hemostasis and disseminated intravascular clotting in Obstetrics

En el período gravido - puerperal, la coagulación sanguínea sufre cambios que se impone conocer para interpretar correctamente esos trastornos cuando se presentan en esta etapa, teniendo en cuenta las posibles complicaciones y el peligro para la vida que pueden presentarse. El objetivo del presente artículo es proporcionar a los obstetras una revisión bibliográfica que les permita actualizarse acerca del tema y facilite su modo de actuación ante pacientes con coagulación intravascular diseminada. Se concluye que es preciso diagnosticar tempranamente el proceso en la fase bioquímica de bajo grado y aplicar en las féminas el tratamiento expedito de la enfermedad de base para eliminar las complicaciones o disminuir su ocurrencia.

In the period from pregnancy to puerperium, blood clotting suffers changes which are important to be known in order to interpret correctly those dysfunctions when they appear in this stage, keeping in mind the possible complications and the danger for life which can take place. The objective of the present work is to provide the obstetricians a literature review that allows them to be updated about the topic and facilitate their performance in case of patients with disseminated intravascular clotting. It is concluded that it is necessary to have an early diagnosis on the process during the low grade biochemical phase and to apply in these women the expedite treatment of the base disease to eliminate complications or to decrease their occurrence.

Whole blood clotting time: Variation of practice in coagulation laboratory, members of Thailand national external quality assessment scheme.

Objective: To collect the necessary data for a perspective of Whole Blood Clotting Time (WBCT) practice in Thailand. Methods: In March 2007, 124 questionnaires were sent to laboratory members of the Thailand National External Quality Assessment Scheme (Thailand NEQAS) to obtain essential information about the WBCT practice. Results: From a dispatch of 124 questionnaires, 120 (96.77%) were returned. There were 101 (84.1%) hospitals performing WBCT in the laboratories and the mean number of WBCTs performed was 16.17 times/month. Eighty nine laboratories (88.11%) used the modified Lee-White methods. Seventy four laboratories (73.26%) used snake bite and other animal bites as the indication for WBCT. Thirty three laboratories (34.37%) had problems performing the WBCT. Conclusion: The WBCT methods among the practice of the Thailand NEQAS laboratory members were as various as problems concerning WBCT throughout Thailand. Their practice needs to be improved and standardized by proper education. It also emphasizes the need for an appropriate guidelines for WBCT in Thailand.

Experimental Assessment of Hemostatic Agents: Comparison with New Developed Chitosan-Based Material

PURPOSE: Many hemostatic agents and dressings have been tested with variable degree of success. Chitosan has a positive charge, it attracts red blood cells, which have a negative charge. Our goal is to test the efficacy of new developed chitosan-based hemostatic materials in providing durable hemostasis in a high-flow arterial wound model. METHODS: We compared each group with SD rats motality tests and in vitro blood compatibility test by blood clotting index (BCI). We devided the SD rats into 6 groups (N =15) by type of hemostatic agents. A: 100% nonwoven chitosan (degree of the deacetylation: 90%). B: 50% N-acetylation on nonwoven of chitosan gel (degree of the deacetylation: 50%). C: 60% N-acetylation on nonwoven of chitosan ge (degree of the deacetylation: 40%)l. D: Cutanplast(R). E: HemCon(R) F: Gauze. In vivo test, a proximal arterial injury was created in unilateral femoral arteries of 90 anesthetized SD rats. Each materials was made same size and thickness then applied to the injury site for 3 minutes. In vitro test, we compared each group with BCI in human blood. RESULTS: In vivo test, group A showed lower motality rate of 46% than any other groups, Group B and C showed lower motality rate of 60% than group D and E's motality rate of 66%. In vitro test, BCI of group A (30.6 +/- 1.2) and B (29.3 +/- 1.0) were showed nearly about group D (29.1 +/- 1.8) and E (27.4 +/- 1.6). Group C (37.1 +/- 2.0) showed higher BCI than group A and B, it means group C decreased blood clotting. CONCLUSION: In conclusion, this study suggests a newly developed chitosan-based hemostatic materials induced durable hemostasis and increased blood clotting, and are considered as effective biologic hemostatic agents.

Clotting mechanism in beagles irradiated by 4.5 Gy γ-rays / 中华放射医学与防护杂志

Objective To explore the clotting mechanism in beagles irradiated by 4.5 Gy γ-rays after treatment with supportive care,or supportive care and combined cytokines.Methods Sixteen beagles were divided into irradiation control group,Supportive care group and combined cytokines treatment group.Platelet aggregation test,thrombelagtography (TEG) and the time measurement were analyzed in vitro.Results In irradiation group and supportive care group,the platelet aggregation rates in beagles were decreased markedly and the k value of TEG was increased 7 d post-irradiation,while those indexes in combined cytokines treatment group changed little.At 14 d post-irradiation,each parameter of TEG in irradiated group changed obviously.The values of r,k,r+k and M were elevated significantly,clotting time and the maximum coagulation time of thrombus delayed,the Ma value was decreased markedly,and the maximum elasticity amplitude of thrombus was diminished.All parameters in combined cytokines treatment group were better than those in supportive care group.The thrombin time was prolonged obviously in irradiated group 14 d post-irradiation,while the thrombin time was the longest at 2-3 weeks post irradiation in supportive care group and combined cytokines treatment group(P>0.05).Conclusions Cytokines could improve the platelet aggregation and the blood clotting functions of beagles suffering from acute radiation sickness.

Procoagulant properties of human MV3 melanoma cells

A correlation between cancer and prothrombotic states has long been described. More recently, a number of studies have focused on the procoagulant mechanisms exhibited by tumor cells. In the present study, we dissected the molecular mechanisms responsible for the procoagulant activity of MV3, a highly aggressive human melanoma cell line. It was observed that tumor cells strongly accelerate plasma coagulation as a result of: i) expression of the blood clotting initiator protein, a tissue factor, as shown by flow cytometry and functional assays (factor Xa formation in the presence of cells and factor VIIa), and ii) direct activation of prothrombin to thrombin by cells, as evidenced by hydrolysis of the synthetic substrate, S-2238, and the natural substrate, fibrinogen. This ability was highly potentiated by the addition of exogenous factor Va, which functions as a co-factor for the enzyme factor Xa. In contrast, prothrombin activation was not observed when cells were previously incubated with DEGR-factor Xa, an inactive derivative of the enzyme. Moreover, a monoclonal antibody against bovine factor Xa reduced the prothrombin-converting activity of tumor cells. In conclusion, the data strongly suggest that MV3 cells recruit factor Xa from the culture medium, triggering an uncommon procoagulant mechanism.