Bothrops Moojeni Snake Venom: A Source of Potential Therapeutic Agents Against Hemostatic Disorders

Abstract Hemostasis is a complex set of biological processes responsible for blood fluidity within normal vessels and for the physiological interruption of bleeding in cases of vascular injury. Bothrops moojeni snake venom is rich in bioactive compounds of pharmacological and clinical interest since its protein components are capable of interfering with many points of the hemostatic process. Here, we present the B. moojeni venom proteins that affect hemostasis and discuss their pharmacological and clinical potential. This systematic review was conducted according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) protocol. Data were obtained from the CAPES Journal Portal database, using the terms "Bothrops" AND "hemostasis", in a search for scientific articles made available in the last 20 years. Many components isolated from B. moojeni snake venom are characterized for their effect on hemostasis and possible application in the diagnosis and treatment of hemostatic disorders.

Mordeduras de ofidios en mano pediátrica y sus complicaciones locales / Snake bites in pediatric hand and their local complications / Mordidas de serpentes de mão pediátricas e suas complicações locais

Introducción: el envenenamiento por mordedura de ofidios es reconocido como un problema de salud pública según la Organización Mundial de la Salud. La baja incidencia sumada a la diversidad de presentaciones clínicas, edades, topografías afectadas, así como los diferentes protocolos en la bibliografía existente sobre algunos aspectos del tratamiento, hacen difícil el manejo sistematizado de estos pacientes. El objetivo de este trabajo es realizar una revisión sistemática de la literatura sobre mordedura de serpientes en pacientes pediátricos con afectación en mano y miembro superior, haciendo hincapié en la conducta frente las complicaciones loco-regionales. Por importancia y frecuencia destacamos al síndrome compartimental, las flictenas y las infecciones. Metodología: se realizó una búsqueda bibliográfica en MedLine/PubMed con las palabras clave: "Snake Bite hand Children" y "Snake Bite compartimental syndrome". Se incluyeron los artículos publicados en los últimos 10 años (2012 al 2022). Resultados: la búsqueda de artículos ante las palabras "Snake Bite hand Children" resultó en 20 articulos y la busqueda ante las palabras "Snake Bite compartimental syndrome" derivó en 34. Luego de aplicar los criterios de inclusión y exclusión se obtuvieron 30 artículos para el análisis. Conclusiones: la población pediátrica se encuentra más expuesta a las mordeduras por serpientes y a su vez a presentar lesiones más severas. El tratamiento del síndrome compartimental continúa siendo un tema de debate. El veneno inoculado puede simular un síndrome compartimental que puede revertir sin fasciotomías con el tratamiento adecuado. Igualmente, ante síntomas y signos claros de síndrome compartimental se sugiere realizar fasciotomías frente a las graves secuelas potenciales. Ante la aparición de flictenas, el destechado cuidadoso de la misma es un tratamiento adecuado. La mayoría de los autores coinciden con el tratamiento profiláctico con antibioticoterapia.

Introduction: Ophidian bite poisoning is recognized as a public health problem by the World Health Organization. The low incidence added to the diversity of clinical presentations, ages, affected topographies, as well as the different protocols in the existing literature on some aspects of treatment, make the systematized management of these patients difficult. The aim of this work is to carry out a systematic review of the literature on snakebite in pediatric patients with hand and upper limb involvement, with emphasis on the management of loco-regional complications. In terms of importance and frequency, we highlight compartment syndrome, phlyctenas and infections. Methodology: a literature search was carried out in MedLine/PubMed with the keywords: "Snake Bite hand Children" and "Snake Bite compartment syndrome". Articles published in the last 10 years (2012 to 2022) were included. Results: the search for articles with the words "Snake Bite hand Children" resulted in 20 articles and the search for the words "Snake Bite compartment syndrome" resulted in 34 articles. After applying the inclusion and exclusion criteria, 30 articles were obtained for the analysis. Conclusions: the pediatric population is more exposed to snake bites and in turn to present more severe lesions. The treatment of compartment syndrome continues to be a subject of debate. Inoculated venom can simulate a compartment syndrome that can be reversed without fasciotomies with proper treatment. Likewise, in the presence of clear symptoms and signs of compartment syndrome, fasciotomies are suggested because of the serious sequelae generated. In the event of the appearance of phlyctenas, careful unroofing of the phlyctenas would be an appropriate treatment. Most authors agree with prophylactic treatment with antibiotic therapy.

Introdução: O envenenamento por mordidas ofídias é reconhecido como um problema de saúde pública pela Organização Mundial da Saúde. A baixa incidência, juntamente com a diversidade de apresentações clínicas, idades, topografias afetadas, bem como os diferentes protocolos da literatura existente sobre alguns aspectos do tratamento, tornam difícil o gerenciamento sistemático desses pacientes. O objetivo deste trabalho é realizar uma revisão sistemática da literatura sobre mordida de cobra em pacientes pediátricos com envolvimento de mãos e membros superiores, com ênfase no gerenciamento de complicações loco-regionais. Em termos de importância e freqüência, destacamos a síndrome compartimental, as flectenas e as infecções. Metodologia: foi realizada uma pesquisa bibliográfica no MedLine/PubMed com as palavras-chave: "Snake Bite hand Children" e "Snake Bite compartment syndrome". Os artigos publicados nos últimos 10 anos (2012 a 2022) foram incluídos. Resultados: a busca de artigos usando as palavras "Snake Bite hand Children" resultou em 20 artigos e a busca das palavras "Snake Bite compartment syndrome" resultou em 34 artigos. Após a aplicação dos critérios de inclusão e exclusão, foram obtidos 30 artigos para análise. Conclusões: a população pediátrica está mais exposta às picadas de cobra e, por sua vez, a lesões mais graves. O tratamento da síndrome compartimental continua a ser motivo de debate. O veneno inoculado pode simular uma síndrome de compartimento que pode ser revertida sem fasciotomias com tratamento apropriado. Da mesma forma, se houver sinais e sintomas claros de síndrome compartimental, são sugeridas fasciotomias por causa das severas seqüelas. Se as flectenas aparecerem, o desenrolamento cuidadoso das flectenas seria um tratamento apropriado. A maioria dos autores concorda com o tratamento profilático com a antibioticoterapia.

Massive acute ischemic stroke after Bothrops spp. envenomation in southwestern Colombia: Case report and literature review / Accidente cerebrovascular isquémico agudo masivo por veneno de Bothrops spp. en el suroccidente de Colombia: reporte de caso y revisión de la literatura

Bothrops spp. envenomation and its relationship with ischemic stroke has complex pathogenesis. Local effects such as edema, pain, redness, necrosis, and systemic manifestations like coagulation disorders, thrombosis, renal failure, and hemorrhage have been reported. Hemorrhagic stroke is a common neurological complication but ischemic stroke is poorly understood. We present here the case of a 50-year-old male with no comorbidities referred from a rural area in southwest Colombia with a Bothrops spp. snakebite on the left hand. On admission, the patient presented with a deterioration of consciousness and required mechanical ventilation assistance. The MRI showed multiple ischemic areas in the bilateral frontal- temporal and occipital regions. Two months later, the patient had a favorable resolution, although central paresis in the III and VI cranial nerves and positive Babinski's sign persisted. As already mentioned, the pathophysiology of ischemic stroke due to snakebite is complex but the procoagulant activity of the venom components, the hypovolemic shock, the endothelial damage, and the thromboinflammation can explain it, and although it rarely occurs, it should be considered as a complication of ophidian accidents caused by Bothrops spp.

La mordedura de serpientes Bothrops spp. y el ataque cerebrovascular isquémico tienen una patogenia compleja. Se reconocen efectos locales como edema, dolor, enrojecimiento y necrosis, así como manifestaciones sistémicas como trastornos de la coagulación, trombosis, insuficiencia renal y hemorragia, por lo que el accidente cerebrovascular hemorrágico es una complicación neurológica común, pero, en cambio, el accidente cerebrovascular isquémico es poco conocido. Se presenta el caso de un paciente de 50 años, sin comorbilidades, remitido de una zona rural del suroccidente de Colombia debido a la mordedura de una serpiente Bothrops spp. en su mano izquierda. En el momento del ingreso, el paciente presentaba deterioro de la conciencia y requirió asistencia respiratoria mecánica. Mediante resonancia magnética, se observaron múltiples áreas isquémicas bilaterales en la región fronto-temporal y en la occipital. Dos meses después, el paciente había evolucionado favorablemente, pero persistían la paresia en los pares craneales III y VI y el signo de Babinski. La fisiopatología del accidente cerebrovascular isquémico por mordedura de serpiente es compleja. La actividad procoagulante de los componentes del veneno, el choque hipovolémico, el daño endotelial y la tromboinflamación pueden explicar el accidente cerebrovascular isquémico que, aunque raro, debe considerarse como una complicación del accidente ofídico causado por serpientes Bothrops spp.

Thrombolysis, clotting, and genotoxic activities modulated by essential oils extracted from Lippia alba

Abstract The composition and pharmacological properties of Lippia alba (Mill.) (L. alba) (Verbenaceae) flower and leaf essential oils (EO) were determined in this study. The major constituents in the flower EO were geranial (49.83%) and neral (32.75%), and in the leaf EO were geranial (38.06%), neral (31.02%), and limonene (18.03%). Flower EO inhibited thrombolysis induced by Bothrops moojeni (B. moojeni) and Lachesis muta muta (L. muta muta) venoms (0.05-1.2 µL mL-1). When tested against L. muta muta venom, the protective effect was smaller in both EO. The EOs prolonged the clotting time induced by L. muta muta venom and a procoagulant effect was observed on B. moojeni. In the comet assay, the flower EO presented anti-genotoxic action (damage frequency of only 11.6 - 34.9%) against the L. muta muta venom. The positive control (Doxorubicin) and the venom alone presented a damage frequency of 80.3% and 70.7%, respectively. The flower EO protected DNA from damage induced by L. muta muta venom. L. alba leaf and flower EOs presented anti-genotoxic action

Profile of snakebite victims reported in a public teaching hospital: a cross-sectional study / Perfil de las víctimas de accidentes ofídicos notificados en un hospital público docente: estudio transversal / Perfil das vítimas de acidente ofídico notificadas em um hospital público de ensino: estudo transversal

ABSTRACT Objective: To understand the epidemiological and clinical profile of snakebite victims in a public teaching hospital. Method: Exploratory, descriptive, cross-sectional study with a quantitative approach conducted in a public hospital in the Triângulo Sul region of Minas Gerais state, Brazil. The data were collected from medical records and compulsory case report forms of patients admitted in the Emergency Service from June 2018 to May 2019. The data were descriptively analyzed and presented in tables. Results: The 137 patients were predominantly male (74.5%), rural workers (33.4%), in their working age (mean of 43.49 years), and in rural areas (87.6%). Bothrops was the most common genus (70.8%), and most occurrences were classified as mild (53.3%). The most performed treatment was snake antivenom administration (73%). Conclusion: Snakebite victims were mostly men in their working age and rural workers. Bothrops bites and mild occurrences were prevalent. Most treatments employed the appropriate snake antivenom. These occurred mostly in rural areas. The prevalent bitten body parts were feet, legs, and hands. Care was provided in less than six hours.

RESUMEN Objetivo: Conocer el perfil epidemiológico y clínico de las víctimas de accidentes ofídicos en un hospital público docente. Método: Estudio exploratorio, descriptivo, transversal, con enfoque metodológico cuantitativo, realizado en un hospital público de la región del Triângulo Sul del estado de Minas Gerais, Brasil. Los datos se recogieron de las historias clínicas y formularios de notificación obligatoria de los pacientes ingresados en los Servicios de Urgencia desde junio de 2018 hasta mayo de 2019. Los datos se analizaron de forma descriptiva y se presentaron en forma de tablas. Resultados: De un total de 137 pacientes, predominaron el sexo masculino (74,5%), los trabajadores rurales (33,4%), los individuos en edad productiva (media de 43,49 años) y la zona de ocurrencia rural (87,6%). El género más implicado en los accidentes fue el Bothrops (70,8%) y la clasificación de la mayoría fue leve (53,3%). El tratamiento más común fue la administración del suero antiofídico (73%). Conclusión: Los accidentes ofídicos se produjeron sobre todo en hombres en edad productiva y en trabajadores rurales. Prevalecieron los accidentes por el género Bothrops y los que se clasifican como leves. La mayoría de los tratamientos utilizaron el antiofídico adecuado. La zona más común de ocurrencia fue la rural. Las zonas corporales más afectadas por las mordeduras fueron los pies, las piernas y las manos, y los tratamientos se realizaron en menos de 6 horas.

RESUMO Objetivo: Conhecer o perfil epidemiológico e clínico das vítimas de acidente ofídico em um hospital público de ensino. Método: Estudo exploratório, descritivo, transversal, de abordagem metodológica quantitativa, realizado em um hospital público do Triângulo Sul de Minas Gerais. A coleta de dados foi realizada em prontuários e fichas de notificação compulsória de pacientes admitidos no Pronto Socorro de junho de 2018 a maio de 2019. Os dados foram analisados descritivamente e apresentados em forma de tabelas. Resultados: De um total de 137 pacientes, foram predominantes o sexo masculino (74,5%), trabalhadores rurais (33,4%), indivíduos em idade produtiva (média de 43,49 anos) e zona de ocorrência rural (87,6%). O gênero mais envolvido nos acidentes foi o Bothrops (70,8%) e a classificação da maioria foi leve (53,3%). O tratamento mais realizado foi a administração do soro antiofídico (73%). Conclusão: Os acidentes ofídicos aconteceram majoritariamente em homens em idade produtiva e trabalhadores rurais. Prevaleceram os acidentes pelo gênero Bothrops e classificados como leves. A maioria dos tratamentos utilizou soro antiofídico adequado. A zona de maior ocorrência foi a rural. As áreas corporais prevalentes de mordedura foram pé, perna e mão e os atendimentos foram realizados em menos de 6 horas.

In vivo and in vitro prospection of the anti-ophidic properties exercised by the extracts of Jacaranda decurrens L

The research and development of alternative treatments for snakebites (e.g., medicinal plants) is necessary due to the high costs of the existing ones. The effects of the aqueous extracts from Jacaranda decurrens leaves, roots, and xylopodium were analyzed upon the venom-induced (Bothrops spp. and Crotalus spp.) systemic and local toxicity. The extracts were able to partially inhibit the phospholipase activity of the venoms from Bothrops jararacussu and Crotalus durissus terrificus. The myotoxic, edema-inducing, coagulant, and hemorrhagic activities were also inhibited. The SDS-PAGE showed that the venom proteins were intact after their incubation with the extracts. This suggests that the possible mechanism of inhibition is not related to the degradation of the protein but rather to their binding to specific sites of the enzymes. The extracts significantly prolonged the survival time of animals in the lethality assay performed with Crotalus durissus terrificus venom and its toxin (crotoxin). The anti-ophidic activity of medicinal plants may aid in the management of snakebites in distant locations by reducing the victim’s local effects and time to heal.

Profile of snakebite victims reported in a public teaching hospital: a cross-sectional study / Perfil de las víctimas de accidentes ofídicos notificados en un hospital público docente: estudio transversal / Perfil das vítimas de acidente ofídico notificadas em um hospital público de ensino: estudo transversal

ABSTRACT Objective: To understand the epidemiological and clinical profile of snakebite victims in a public teaching hospital. Method: Exploratory, descriptive, cross-sectional study with a quantitative approach conducted in a public hospital in the Triângulo Sul region of Minas Gerais state, Brazil. The data were collected from medical records and compulsory case report forms of patients admitted in the Emergency Service from June 2018 to May 2019. The data were descriptively analyzed and presented in tables. Results: The 137 patients were predominantly male (74.5%), rural workers (33.4%), in their working age (mean of 43.49 years), and in rural areas (87.6%). Bothrops was the most common genus (70.8%), and most occurrences were classified as mild (53.3%). The most performed treatment was snake antivenom administration (73%). Conclusion: Snakebite victims were mostly men in their working age and rural workers. Bothrops bites and mild occurrences were prevalent. Most treatments employed the appropriate snake antivenom. These occurred mostly in rural areas. The prevalent bitten body parts were feet, legs, and hands. Care was provided in less than six hours.

RESUMEN Objetivo: Conocer el perfil epidemiológico y clínico de las víctimas de accidentes ofídicos en un hospital público docente. Método: Estudio exploratorio, descriptivo, transversal, con enfoque metodológico cuantitativo, realizado en un hospital público de la región del Triângulo Sul del estado de Minas Gerais, Brasil. Los datos se recogieron de las historias clínicas y formularios de notificación obligatoria de los pacientes ingresados en los Servicios de Urgencia desde junio de 2018 hasta mayo de 2019. Los datos se analizaron de forma descriptiva y se presentaron en forma de tablas. Resultados: De un total de 137 pacientes, predominaron el sexo masculino (74,5%), los trabajadores rurales (33,4%), los individuos en edad productiva (media de 43,49 años) y la zona de ocurrencia rural (87,6%). El género más implicado en los accidentes fue el Bothrops (70,8%) y la clasificación de la mayoría fue leve (53,3%). El tratamiento más común fue la administración del suero antiofídico (73%). Conclusión: Los accidentes ofídicos se produjeron sobre todo en hombres en edad productiva y en trabajadores rurales. Prevalecieron los accidentes por el género Bothrops y los que se clasifican como leves. La mayoría de los tratamientos utilizaron el antiofídico adecuado. La zona más común de ocurrencia fue la rural. Las zonas corporales más afectadas por las mordeduras fueron los pies, las piernas y las manos, y los tratamientos se realizaron en menos de 6 horas.

RESUMO Objetivo: Conhecer o perfil epidemiológico e clínico das vítimas de acidente ofídico em um hospital público de ensino. Método: Estudo exploratório, descritivo, transversal, de abordagem metodológica quantitativa, realizado em um hospital público do Triângulo Sul de Minas Gerais. A coleta de dados foi realizada em prontuários e fichas de notificação compulsória de pacientes admitidos no Pronto Socorro de junho de 2018 a maio de 2019. Os dados foram analisados descritivamente e apresentados em forma de tabelas. Resultados: De um total de 137 pacientes, foram predominantes o sexo masculino (74,5%), trabalhadores rurais (33,4%), indivíduos em idade produtiva (média de 43,49 anos) e zona de ocorrência rural (87,6%). O gênero mais envolvido nos acidentes foi o Bothrops (70,8%) e a classificação da maioria foi leve (53,3%). O tratamento mais realizado foi a administração do soro antiofídico (73%). Conclusão: Os acidentes ofídicos aconteceram majoritariamente em homens em idade produtiva e trabalhadores rurais. Prevaleceram os acidentes pelo gênero Bothrops e classificados como leves. A maioria dos tratamentos utilizou soro antiofídico adequado. A zona de maior ocorrência foi a rural. As áreas corporais prevalentes de mordedura foram pé, perna e mão e os atendimentos foram realizados em menos de 6 horas.

Caracterización bioquímica del veneno de la serpiente Bothrops roedingeri Mertens, 1942, y sus actividades edematógena, hemorrágica y miotóxica / Biochemical characterization of Bothrops roedingeri Mertens, 1942 snake venom and its edematogenic, hemorrhagic, and myotoxic activities

Resumen : Introducción. El envenenamiento por mordedura de serpiente es considerado por la Organización Mundial de la Salud (OMS) una enfermedad tropical desatendida. Si bien los venenos de otras serpientes Bothrops se vienen estudiado ampliamente, poco se conoce del de Bothrops roedingeri. Objetivos. Caracterizar bioquímicamente el veneno total de la serpiente B. roedingeri y evaluar su actividad miotóxica, edematógena y hemorrágica. Materiales y métodos. Se hizo la caracterización enzimática del veneno de B. roedingeri determinando la actividad de la fosfolipasa A2 (PLA2) y de las enzimas proteolíticas, así como su acción fibrinogenolítica mediante electroforesis en gel de poliacrilamida con duodecilsulfato sódico (sodium dodecyl sulfate polyacrylamide gel electrophoresis, SDS-PAGE), y la caracterización tóxica del veneno estableciendo la dosis hemorrágica mínima, la dosis edematógena mínima y el efecto miotóxico local y sistémico. Resultados. La actividad de las PLA2 del veneno total de B. roedingeri fue de 3,45 ± 0,11 nmoles/minuto, la proteolítica, de 0,145 ± 0,009 nmoles/minuto, en tanto que el índice de coagulación del fibrinógeno fue de 6,67 ± 1,33 segundos. Por otro lado, el veneno produjo una dosis hemorrágica mínima de 24,5 µg, una dosis edematógena mínima de 15,6 µg y un pronunciado efecto miotóxico local evidenciado por la elevación de los niveles plasmáticos de creatina cinasa después de la inoculación por vía intramuscular. No se registró miotoxicidad sistémica. Conclusiones. El veneno de B. roedingeri tiene efectos hemorrágicos, edematógenos y miotóxicos locales, así como una elevada actividad de la PLA2, que sería responsable de los efectos miotóxico y edematógeno. También presentó actividad proteolítica, la cual podría afectar la coagulación, dada su capacidad para degradar el fibrinógeno y producir hemorragia por acción de las metaloproteasas.

Abstract : Introduction: Snakebite envenoming is considered by the World Health Organization (WHO) as a neglected tropical disease. Currently, Bothrops snake venoms are being studied intensively, but there is little knowledge about Bothrops roedingeri venom. Objectives: To biochemically characterize B. roedingeri total venom and evaluate its myotoxic, edematogenic, and hemorrhagic activity. Materials and methods: We characterized B. roedingeri venom enzymatic activity by determining the phospholipase A2 and the proteolytic and fibrinogenolytic action using SDS-PAGE electrophoresis while we characterized its venom toxicity by determining the minimum hemorrhagic dose, the minimum edema dose, and the local and systemic myotoxic effects. Results: Bothrops roedingeri venom showed a PLA2 activity of 3.45 ± 0.11 nmoles/min, proteolytic activity of 0.145 ± 0.009 nmoles/min, and a fibrinogen coagulation index of 6.67 ± 1.33 seconds. On the other hand, it produced an minimum hemorrhagic dose of 24.5 µg, an minimum edema dose of 15.6 µg, and a pronounced local myotoxic effect evidenced by the elevation of plasma creatine kinase levels after intramuscular inoculation. The venom showed no systemic myotoxicity. Conclusions: Bothrops roedingeri venom has local hemorrhagic, edematogenic, and myotoxic activity. Enzymatically, it has high PLA2 activity, which would be responsible for the myotoxic and edematogenic effects. It also has proteolytic activity, which could affect coagulation given its ability to degrade fibrinogen, and it causes bleeding through the metalloproteases.

Effect of Bitis gabonica and Dendroaspis angusticeps snake venoms on apoptosis-related genes in human thymic epithelial cells

Certain environmental toxins permanently damage the thymic epithelium, accelerate immune senescence and trigger secondary immune pathologies. However, the exact underlying cellular mechanisms and pathways of permanent immune intoxication remain unknown. The aim of the present study was to demonstrate gene expressional changes of apoptosis-related cellular pathways in human thymic epithelial cells following exposure to snake venom from Bitis gabonica and Dendroaspis angusticeps. Methods: Snake venoms were characterized by analytical methods including reversed phase high-performance liquid chromatography and sodium dodecyl sulphate-polyacrylamide gel electrophoresis, then applied on human thymic epithelial cells (1889c) for 24 h at 10 μg/mL (as used in previous TaqMan Array study). Gene expressional changes restricted to apoptosis were assayed by TaqMan Array (Human Apoptosis Plate). Results: The most prominent gene expressional changes were shown by CASP5 (≈ 2.5 million-fold, confirmed by dedicated quantitative polymerase chain reaction) and CARD9 (0.016-fold) for B. gabonica, and BIRC7 (6.46-fold) and CASP1 (0.30-fold) for D. angusticeps. Conclusion: The observed apoptotic environment suggests that pyroptosis may be the dominant pathway through which B. gabonica and D. angusticeps snake venoms trigger thymic epithelial apoptosis following envenomation.(AU)

Oral Immunization for Anti-snake Venom Production in Equine Animals

The immunization process of current commercial manufacturing of anti-snake venom (ASV), uses injections of bentonite, complete Freund’s adjuvant, or incomplete Freund’s adjuvant, mixed with low doses of the snake venom in horses (but rarely in other large mammals), which frequently cause serious adverse effects in host animals. At the site of injection, horses may develop painful swelling, granuloma, abscess, scar, or systemic neurological and hematological defects, low antibody response, or death due to anaphylactic shock. We sought to investigate a novel alternate immunization strategy with oral administration of snake venom with adjuvants. We utilized M5904 mineral oil emulsion as an adjuvant that was mixed with sub-lethal doses (LD) of the snake venoms. Our preliminary experiments were initiated in March 2011 and the present data culminated in March 2018. In our initial experiments which were carried out in inbred mice, the LD100 was 10.36 ug/25 grams of mice for Naja. oxins and 10.0 ug/25 gram of Naja. karachians. We extrapolated the sub-LD dose to horses by cutting the LD100 in mice to 20%. This dose did not cause any apparent pathology in horses and therefore, we adopted that dose for the equine.

Antivenomic approach of different Crotalus durissus collilineatus venoms

Our group has previously performed a proteomic study verifying that individual variations can occur among Crotalus durissus collilineatus venoms. These variations may lead to differences in venom toxicity and may result in lack of neutralization of some components by antivenom. In this way, this study aimed to evaluate the Brazilian anticrotalic serum capacity in recognizing twenty-two Crotalus durissus collilineatus venoms, as well as their fractions. Methods: The indirect enzyme-linked immunosorbent assay (ELISA) was chosen to evaluate the efficacy of heterologous anticrotalic serum produced by Instituto Butantan (Brazil) in recognizing the twenty-two Crotalus durissus collilineatus venoms and the pool of them. Moreover, the venom pool was fractionated using reversed-phase fast protein liquid chromatography (RP-FPLC) and the obtained fractions were analyzed concerning antivenom recognition. Results: Evaluation of venom variability by ELISA showed that all venom samples were recognized by the Brazilian anticrotalic antivenom. However, some particular venom fractions were poorly recognized. Conclusion: This study demonstrated that the Brazilian anticrotalic serum recognizes all the different twenty-two venoms of C. d. collilineatus and their fractions, although in a quantitatively different way, which may impact the effectiveness of the antivenom therapy. These results confirm the need to use a pool of venoms with the greatest possible variability in the preparation of antivenoms, in order to improve their effectiveness.(AU)

Research progress of the clinical hemostatic effect of heamocoagulase agkistrodon / 中国基层医药

Haemocoagulase agkistrodon for injection is a kind of protein hemostatic which is purified from snake venom of Agkistrodon acutus , the hemostatic mechanism of the drug is it that the removal of fibrinogen from fibrinogen in blood can make it into monomeric fibrin after entering the body and enhance coagulation function . Preclinical results showed that the drug can significantly shortened the time of tail bleeding in mice and coagulation time of whole blood in rabbits .Clinical studies show that the medicine has a safe and effective hemostatic effect on incision in abdominal,thyroid,department of obstetrics and gynecology ,department of urology,and the hemostatic effect of thoracic incision is not obvious .In this review,the research progress on the efficacy and safety of Agkistrodon acutus thrombin in clinical hemostasis were reviewed .

Strategies in 'snake venomics' aiming at an integrative view of compositional, functional, and immunological characteristics of venoms

This work offers a general overview on the evolving strategies for the proteomic analysis of snake venoms, and discusses how these may be combined through diverse experimental approaches with the goal of achieving a more comprehensive knowledge on the compositional, toxic, and immunological characteristics of venoms. Some recent developments in this field are summarized, highlighting how strategies have evolved from the mere cataloguing of venom components (proteomics/venomics), to a broader exploration of their immunological (antivenomics) and functional (toxicovenomics) characteristics. Altogether, the combination of these complementary strategies is helping to build a wider, more integrative view of the life-threatening protein cocktails produced by venomous snakes, responsible for thousands of deaths every year.(AU)

Strategies in snake venomics aiming at an integrative view of compositional, functional, and immunological characteristics of venoms

Abstract This work offers a general overview on the evolving strategies for the proteomic analysis of snake venoms, and discusses how these may be combined through diverse experimental approaches with the goal of achieving a more comprehensive knowledge on the compositional, toxic, and immunological characteristics of venoms. Some recent developments in this field are summarized, highlighting how strategies have evolved from the mere cataloguing of venom components (proteomics/venomics), to a broader exploration of their immunological (antivenomics) and functional (toxicovenomics) characteristics. Altogether, the combination of these complementary strategies is helping to build a wider, more integrative view of the life-threatening protein cocktails produced by venomous snakes, responsible for thousands of deaths every year.

Reconhecimento de frações do veneno de cobras corais brasileiras pelo soro antielapídico / Recognition of venom fractions from Brazilian coral snakes by antielapid serum

Objetivos: Investigar a capacidade do soro antielapídico produzido no Brasil na identificação de frações do veneno de seis espécies, incluindo as que constituem o pool de inoculação: Micrurus brasiliensis, M. corallinus, M. frontalis, M. lemniscatus, M. spixii e M. surinamensis. Métodos: As amostras utilizadas fazem parte do banco de venenos do Centro de Estudos e Pesquisas Biológicas, da Pontifícia Universidade Católica de Goiás. O soro antielapídico foi cedido pela Fundação Ezequiel Dias. As metodologias empregadas foram eletroforese e imunoblotting. Resultados: Foi demonstrada uma variabilidade toxinológica e uma capacidade também variável de reconhecimento desses componentes pelo soro antielapídico. A partir da técnica de western-blotting o soro antielapídico da Fundação Ezequiel Dias foi capaz de reconhecer a maioria, mas não todos os componentes presentes nos venenos analisados. Conclusões: Os resultados sugerem uma eficácia restrita do soro antielapídico já que o mesmo possui limitações quanto as espécies Amazônicas, o que reforça a necessidade de uma revisão dos estudos intra e interespecíficos dos venenos micrúricos.

Aims: To investigate whether the antielapid serum produced in Brazil could identify venom fractions from six species of Micrurus, including those in the inoculation pool: Micrurus brasiliensis, M. corallinus, M. frontalis, M. lemniscatus, M. spixii, and M. surinamensis. Methods: The samples belong to the venom bank of the Center for Biological Studies and Research from the Pontifícia Universidade Católica de Goiás, Brazil. The antielapid serum was granted by the Ezequiel Dias Foundation. Both electrophoresis and immunoblotting methods were used. Results: Variability in venom components and in the ability to recognize such components was demonstrated by the antielapid serum. Based on the western-blotting technique, the antielapid serum from Ezequiel Dias Foundation was able to recognize most, but not all the components present in the analyzed venoms. Conclusions: The results suggest restricted efficacy of the antielapid serum, due to its limitations against species from the Amazon region, reinforcing the need for a review of intraspecific and interspecific studies of Micrurus venoms.

Purification procedure for the isolation of a P-I metalloprotease and an acidic phospholipase A 2 fromBothrops atrox snake venom

Background Snake venoms are complex mixtures of inorganic and organic components, mainly proteins and peptides. Standardization of methods for isolating bioactive molecules from snake venoms is extremely difficult due to the complex and highly variable composition of venoms, which can be influenced by factors such as age and geographic location of the specimen. Therefore, this study aimed to standardize a simple purification methodology for obtaining a P-I class metalloprotease (MP) and an acidic phospholipase A2 (PLA 2 ) from Bothrops atroxvenom, and biochemically characterize these molecules to enable future functional studies.Methods To obtain the toxins of interest, a method has been standardized using consecutive isolation steps. The purity level of the molecules was confirmed by RP-HPLC and SDS-PAGE. The enzymes were characterized by determining their molecular masses, isoelectric points, specific functional activity and partial amino acid sequencing.Results The metalloprotease presented molecular mass of 22.9 kDa and pI 7.4, with hemorrhagic and fibrin(ogen)olytic activities, and its partial amino acid sequence revealed high similarity with other P-I class metalloproteases. These results suggest that the isolated metalloprotease is Batroxase, a P-I metalloprotease previously described by our research group. The phospholipase A 2 showed molecular mass of 13.7 kDa and pI 6.5, with high phospholipase activity and similarity to other acidic PLA2 s from snake venoms. These data suggest that the acidic PLA2 is a novel enzyme from B. atrox venom, being denominated BatroxPLA 2 .Conclusions The present study successfully standardized a simple methodology to isolate the metalloprotease Batroxase and the acidic PLA 2 BatroxPLA2 from the venom of B. atrox, consisting mainly of classical chromatographic processes. These two enzymes will be used in future studies to evaluate their effects on the complement system and the inflammatory process, in addition to the thrombolytic potential of the metalloprotease.(AU)

Purification procedure for the isolation of a P-I metalloprotease and an acidic phospholipase A2 from Bothrops atrox snake venom

Background Snake venoms are complex mixtures of inorganic and organic components, mainly proteins and peptides. Standardization of methods for isolating bioactive molecules from snake venoms is extremely difficult due to the complex and highly variable composition of venoms, which can be influenced by factors such as age and geographic location of the specimen. Therefore, this study aimed to standardize a simple purification methodology for obtaining a P-I class metalloprotease (MP) and an acidic phospholipase A2 (PLA 2 ) from Bothrops atroxvenom, and biochemically characterize these molecules to enable future functional studies.Methods To obtain the toxins of interest, a method has been standardized using consecutive isolation steps. The purity level of the molecules was confirmed by RP-HPLC and SDS-PAGE. The enzymes were characterized by determining their molecular masses, isoelectric points, specific functional activity and partial amino acid sequencing.Results The metalloprotease presented molecular mass of 22.9 kDa and pI 7.4, with hemorrhagic and fibrin(ogen)olytic activities, and its partial amino acid sequence revealed high similarity with other P-I class metalloproteases. These results suggest that the isolated metalloprotease is Batroxase, a P-I metalloprotease previously described by our research group. The phospholipase A 2 showed molecular mass of 13.7 kDa and pI 6.5, with high phospholipase activity and similarity to other acidic PLA2 s from snake venoms. These data suggest that the acidic PLA2 is a novel enzyme from B. atrox venom, being denominated BatroxPLA 2 .Conclusions The present study successfully standardized a simple methodology to isolate the metalloprotease Batroxase and the acidic PLA 2 BatroxPLA2 from the venom of B. atrox, consisting mainly of classical chromatographic processes. These two enzymes will be used in future studies to evaluate their effects on the complement system and the inflammatory process, in addition to the thrombolytic potential of the metalloprotease.

Applications of snake venoms in treatment of cancer

Snake venoms are folk medicines used since ages. The components of snake venoms have high specific affinity and actions on cells and cell components. Also snake venoms are largely cytotoxic to tumor cells than normal cells. In addition to these, they have several therapeutic actions that make them an attractive option in the management of cancer. The advent of modern technologies has greatly helped in extracting and identifying new components of therapeutic interests in short time. The article highlights the importance of snake venoms in the management of cancer, so as to motivate curious researchers to devote their skills in this fascinating area. This in turn may bring hope, smile and relief to several cancer patients in future.

Snake venom L-amino acid oxidases: an overview on their antitumor effects

The L-amino acid oxidases (LAAOs) constitute a major component of snake venoms and have been widely studied due to their widespread presence and various effects, such as apoptosis induction, cytotoxicity, induction and/or inhibition of platelet aggregation, hemorrhage, hemolysis, edema, as well as antimicrobial, antiparasitic and anti-HIV activities. The isolated and characterized snake venom LAAOs have become important research targets due to their potential biotechnological applications in pursuit for new drugs of interest in the scientific and medical fields. The current study discusses the antitumor effects of snake venom LAAOs described in the literature to date, highlighting the mechanisms of apoptosis induction proposed for this class of proteins.

Comparative analysis of the venom proteome of four important Malaysian snake species

Naja kaouthia, Ophiophagus hannah, Bungarus fasciatus and Calloselasma rhodostoma are four venomous snakes indigenous to Malaysia. In the present study, their proteomic profile by two-dimensional gel electrophoresis (2-DE) have been separated and compared.