De novo venom gland transcriptomics of Calliophis bivirgata flaviceps: uncovering the complexity of toxins from the Malayan blue coral snake

The Malayan blue coral snake, Calliophis bivirgata flaviceps, is a medically important venomous snake in Southeast Asia. However, the complexity and diversity of its venom genes remain little explored. Methods: To address this, we applied high-throughput next-generation sequencing to profile the venom gland cDNA libraries of C. bivirgata flaviceps. The transcriptome was de novo assembled, followed by gene annotation, multiple sequence alignment and analyses of the transcripts. Results: A total of 74 non-redundant toxin-encoding genes from 16 protein families were identified, with 31 full-length toxin transcripts. Three-finger toxins (3FTx), primarily delta-neurotoxins and cardiotoxin-like/cytotoxin-like proteins, were the most diverse and abundantly expressed. The major 3FTx (Cb_FTX01 and Cb_FTX02) are highly similar to calliotoxin, a delta-neurotoxin previously reported in the venom of C. bivirgata. This study also revealed a conserved tyrosine residue at position 4 of the cardiotoxin-like/cytotoxin-like protein genes in the species. These variants, proposed as Y-type CTX-like proteins, are similar to the H-type CTX from cobras. The substitution is conservative though, preserving a less toxic form of elapid CTX-like protein, as indicated by the lack of venom cytotoxicity in previous laboratory and clinical findings. The ecological role of these toxins, however, remains unclear. The study also uncovered unique transcripts that belong to phospholipase A2 of Groups IA and IB, and snake venom metalloproteinases of PIII subclass, which show sequence variations from those of Asiatic elapids. Conclusion: The venom gland transcriptome of C. bivirgata flaviceps from Malaysia was de novo assembled and annotated. The diversity and expression profile of toxin genes provide insights into the biological and medical importance of the species.(AU)

Landscape of research, production, and regulation in venoms and antivenoms: a bibliometric analysis / Panorama de la investigación, la producción y la regulación de venenos y antivenenos: un análisis bibliométrico / Panorama de pesquisa, produção e regulação de venenos e antivenenos: análise bibliométrica

ABSTRACT Objectives. To assess the productivity and visibility in research, clinical studies, treatment, use and production of antivenoms against poisonous snakes, scorpions and spiders. Methods. Bibliometric analysis of research and other activities. Articles on venoms and antivenoms published between 2000 and 2020 were retrieved from the Scopus database. The records were analyzed by bibliometric indicators including number of documents per year, journals, authors, and citation frequency. VOSviewer® v.1.6.13 was used to construct bibliometric networks for country co-authorships and co-occurrence of terms. Results. Australia, Brazil, Costa Rica and India were among the six top countries with most documents and were selected for more detailed analysis. Costa Rica was the country with the largest percentage of its publications dedicated to antivenom production and venomics. Only a few papers dealt with the issues of quality, safety, and efficacy of antivenoms or the role of the national regulatory authorities. The use of VOSviewer® allowed visualization through joint publications of networking between countries. Visualization by co-occurrence of terms showed differences in the research carried out. Conclusions. Working in a collaborative and coordinated manner these four countries could have a major impact on envenoming globally. Attention should be given not only to antivenom production but also to strengthening regulatory oversight of antivenom products.

RESUMEN Objetivos. Evaluar la productividad y la visibilidad en la investigación, los estudios clínicos, el tratamiento, el uso y la producción de antivenenos contra las picaduras de serpientes, arañas y escorpiones venenosos. Métodos. Análisis bibliométrico de la investigación y de las otras actividades. Se tomaron los artículos sobre venenos y antivenenos publicados entre el 2000 y el 2020 en la base de datos de Scopus. Estos documentos se analizaron mediante indicadores bibliométricos como el número de documentos por año, revistas, autores o frecuencia en las citas. Se utilizó VOSviewer® v.1.6.13 para crear una red bibliométrica para coautorías de países y coapariciones de términos. Resultados. Australia, Brasil, Costa Rica e India estaban entre los seis primeros países con más documentos y se seleccionaron para un análisis más detallado. Costa Rica fue el país con el mayor porcentaje de sus publicaciones dedicadas a la producción de antivenenos y la venómica. Solo unos pocos artículos trataban los temas de la calidad, la seguridad y la eficacia de los antivenenos, o la función de las autoridades regulatorias nacionales. Gracias a VOSviewer® pudimos visualizar las publicaciones conjuntas de las colaboraciones entre países. La visualización por la coaparición de términos arrojó diferencias en la investigación realizada. Conclusiones. Si estos cuatro países trabajasen de forma colaborativa y coordinada, podrían tener una repercusión mayor en los envenenamientos por picaduras en el mundo. El foco no debe ponerse solo en la producción de antivenenos, sino también en fortalecer la supervisión regulatoria de estos productos.

RESUMO Objetivos. Avaliar a produtividade e visibilidade em pesquisa, estudos clínicos, tratamento, uso e produção de antivenenos contra peçonhas de serpentes, escorpiões e aranhas. Métodos. Análise bibliométrica de pesquisas e outras atividades. Artigos sobre venenos e antivenenos publicados entre 2000 e 2020 foram obtidos da base de dados Scopus. O conteúdo foi analisado segundo indicadores bibliométricos, como número de artigos por ano, periódicos, autores e frequência de citação. Utilizou-se o software VOSviewer® v.1.6.13 para construir redes bibliométricas de coautoria de países e co-ocorrência de termos. Resultados. Austrália, Brasil, Costa Rica e Índia figuraram entre os seis principais países com o maior número de artigos e, assim, foram selecionados para uma análise mais aprofundada. A Costa Rica teve a maior porcentagem de publicações dedicadas à produção de antivenenos e pesquisa em venômica. Apenas um pequeno número de artigos tratou de questões relacionadas à qualidade, segurança e eficácia dos antivenenos ou ao papel das autoridades nacionais reguladoras. O software VOSviewer® permitiu visualizar, através das publicações conjuntas, as redes formadas entre diferentes países. A visualização por co-ocorrência de termos revelou diferenças nas pesquisas realizadas. Conclusões. Trabalhando de forma colaborativa e coordenada, esses quatro países tiveram uma influência importante em nível mundial no campo de acidentes por animais peçonhentos. Deve-se atentar não apenas à produção de antivenenos, mas também ao fortalecimento da fiscalização regulatória destes.

Molecular design, pharmacology and toxicology optimization of antimicrobial peptide from Hydrophis cyanocinctus, Hc-CATH / 生物工程学报

Based on the cathelicidin family antimicrobial peptide Hc-CATH derived from sea snake, the Hc-16 and Hc-15 of 16 and 15 amino acid residues, were designed. By using CCK8, minimal inhibitory concentration, ELISA and bio-layer interferometry assays, their cytotoxicity, antibacterial activity, anti-inflammatory activity, and LPS neutralization activity was examined. Compared with Hc-15, Hc-16 had lower cytotoxicity and better broad-spectrum antibacterial activity against pathogens including clinically resistant bacteria, with the minimum inhibitory concentration of only 4.69 μg/mL. Hc-16 inhibited the expression of inflammatory cytokines of TNF-α and IL-6 induced by LPS, so as to significantly reduce the inflammatory response induced by infection. In addition, structure-activity relationship studies have shown that the phenylalanine at the C- and N-terminals of Hc-16 played a crucial role in its antibacterial and anti-inflammatory activity. Altogether, the designed Hc-16 has an excellent prospect to be developed into a novel antibiotic.

Identification of Chinese medicine sea snake based on cytochrome B (Cytb) / 中国中药杂志

The identification of species primordium has been one of the hot issues in the identification of traditional Chinese medicine. Sea snake is one of the most valuable Chinese medicinal materials in China. In order to understand the origin and varieties of sea snake in the market, we studied the molecular identification of 46 sea snakes by cytochrome B(Cytb). After comparison and manual correction, the sequence length was 582 bp, and the content of A+T(58.9%) was higher than that of G+C(41.1%). There exist 197 variable sites and 179 parsimony-informative sites of the sequence. There are 44 kinds of sequence alignment with consistency equal to 100%, and 2 kinds equal to 96%. A total of 408 Cytb effective sequences were downloaded from GenBank database, with a total of 68 species. Phylogenetic tree of a total of 454 sea snake sequences with the samples in this study were constructed by neighbor-joining trees and Bayesian inference method, respectively, which can identify 42 samples of medicinal materials, while 4 samples can not be identified because of their low node support. The results showed that the species of the sea snake medicine were at least from 2 genera and 5 species, namely, Aipysurus eydouxii, Hydrophis curtus, H. caerulescen, H. curtus, H. ornatus and H. spiralis. This study suggested that the original species of commercial sea snake are very complex and can provide insight into the identification of sea snakes.

Neurotoxical activity of Micrurus snake venom and methods for its analysis. A literature review / Actividad neurotóxica del veneno de serpientes del género Micrurus y métodos para su análisis. Revisión de la literatura

Abstract Introduction: Snakes of the genus Micrurus have fossorial habits, passive temperament and scarce production of powerful venom with neurotoxic characteristics that block the synaptic transmission at the neuromuscular junction. Objective: To present an overview of the neurotoxicity of the Micrurus snake venom, and its functional characterization by ex vivo analysis methods. Materials and methods: A literature review was conducted in MedLine and ScienceDirect using specific terms and their combinations. Search strategy: type of studies: articles on the neurotoxicity of Micrurus snake venom and techniques to determine its neurotoxic activity by in vitro, in vivo and ex vivo models; publication period: articles published until June 2018; publication language: English and Spanish. Results: Out of 88 studies identified in the initial search, 28 were excluded because they did not meet the inclusion criteria (based on reading their titles and abstracts). 8 additional articles (books and reports) were included, since, according to the authors' opinion, they complemented the information reported by the selected studies. The studies included in the review (n=68) were original research papers (n=44), review articles (n = 16), and book chapters, reports, guides and online consultations (n=8). Conclusions: Studies performed using ex vivo muscle and nerve preparations to evaluate the effect of neurotoxins provide a good model for the characterization of the pre-synaptic and post-synaptic effect of the venom produced by snakes of the genus Micrurus.

Resumen Introducción. Las serpientes del género Micrurus son animales de hábitos fosoriales, de temperamento pasivo y escasa producción de un potente veneno con características neurotóxicas que bloquean la transmisión sináptica en la placa neuromuscular. Objetivo. Presentar un panorama general de la neurotoxicidad del veneno de las serpientes Micrurus y su caracterización funcional mediante métodos de análisis ex vivo. Materiales y métodos. Se realizó una revisión de la literatura en MedLine y ScienceDirect usando términos específicos y sus combinaciones. Estrategia de búsqueda: tipo de estudios: artículos sobre la neurotoxicidad del veneno de serpientes Micrurus y técnicas para determinar su actividad neurotóxica mediante modelos in vitro, in vivo y ex vivo; periodo de publicación: sin límite inicial a junio de 2018; idiomas: inglés y español. Resultados. De los 88 estudios identificados en la búsqueda inicial, se excluyeron 28 por no cumplir los criterios de inclusión (basándose en la lectura de títulos y resúmenes); además, se incluyeron 8 documentos adicionales (libros e informes), que, a criterio de los autores, complementaban la información reportada por las referencias seleccionadas. Los estudios incluidos en la revisión (n=68) correspondieron a las siguientes tipologías: investigaciones originales (n=44), artículos de revisión (n=16) y capítulos de libros, informes, guías y consultas en internet (n=8). Conclusiones. Los estudios que describen el uso de preparaciones ex vivo de músculo y nervio para evaluar el efecto de neurotoxinas ofrecen un buen modelo para la caracterización del efecto presináptico y postsináptico del veneno producido por las serpientes Micrurus.

A Kunitz-type peptide from Dendroaspis polylepis venom as a simultaneous inhibitor of serine and cysteine proteases

Proteases play an important role for the proper physiological functions of the most diverse organisms. When unregulated, they are associated with several pathologies. Therefore, proteases have become potential therapeutic targets regarding the search for inhibitors. Snake venoms are complex mixtures of molecules that can feature a variety of functions, including peptidase inhibition. Considering this, the present study reports the purification and characterization of a Kunitz-type peptide present in the Dendroaspis polylepis venom as a simultaneous inhibitor of elastase-1 and cathepsin L. Methods: The low molecular weight pool from D. polylepis venom was fractionated in reverse phase HPLC and all peaks were tested in fluorimetric assays. The selected fraction that presented inhibitory activity over both proteases was submitted to mass spectrometry analysis, and the obtained sequence was determined as a Kunitz-type serine protease inhibitor homolog dendrotoxin I. The molecular docking of the Kunitz peptide on the elastase was carried out in the program Z-DOCK, and the program RosettaDock was used to add hydrogens to the models, which were re-ranked using ZRANK program. Results: The fraction containing the Kunitz molecule presented similar inhibition of both elastase-1 and cathepsin L. This Kunitz-type peptide was characterized as an uncompetitive inhibitor for elastase-1, presenting an inhibition constant (Ki) of 8 μM. The docking analysis led us to synthesize two peptides: PEP1, which was substrate for both elastase-1 and cathepsin L, and PEP2, a 30-mer cyclic peptide, which showed to be a cathepsin L competitive inhibitor, with a Ki of 1.96 µM, and an elastase-1 substrate. Conclusion: This work describes a Kunitz-type peptide toxin presenting inhibitory potential over serine and cysteine proteases, and this could contribute to further understand the envenomation process by D. polylepis. In addition, the PEP2 inhibits the cathepsin L activity with a low inhibition constant.(AU)

Trastorno de excoriación en una mujer con panarteritis nudosa. Reporte de caso / Skin Picking Disorder in a Woman with Polyarteritis Nodosa. Case Report

RESUMEN Introducción: El trastorno por excoriación está incluido en el DSM - 5 dentro de la categoría Recibido el 4 de abril de 2017 de trastorno obsesivo compulsivo y trastornos relacionados. Se define como la urgencia Aceptado el 18 de marzo de 2018 de tocar, rascar, frotar, restregar, friccionar, apretar, morder o excavar la piel de forma On-line el 7 de mayo de 2018 recurrente hasta producirse lesiones cutáneas. Es un trastorno poco frecuente (1.4 - 5.4% de la población) y se presenta principalmente en mujeres. Presentación de caso: Se presenta el caso de una mujer de 31 años quien fue valorada por dermatología y ortopedia por presencia de lesiones ulceradas e infectadas en miembros inferiores, junto con otras lesiones superficiales por rascado en tórax, brazos, antebrazos, espalda y cabeza; además reportando síntomas ansiosos, razón por la cual es valorada por el servicio de Psiquiatría de enlace Discusión: El rascado cutáneo, conducta normal en los mamíferos, cobra valor patológico desde el punto de vista psiquiátrico al ser un acto repetitivo y persistente, como la conducta que se presenta en el trastorno por excoriación. Dada la relación descrita con el espectro obsesivo - compulsivo, se recomienda el uso de inhibidores selectivos de la recaptación de serotonina y la terapia cognitivo conductual.

ABSTRACT Introduction: Excoriation (skin picking) disorder is included in the DSM-5 in the obsessive compulsive and related disorders category. It is defined as the recurrent urge to touch, scratch, scrape, scrub, rub, squeeze, bite or dig in the skin, leading to skin lesions. It is a rare disorder (1.4-5.4% of the population) and occurs mainly in women. Case report: this article reports the case of a 31-year-old female patient, initially assessed by dermatology and orthopaedics for the presence of infected ulcerated lesions on her lower limbs, with other superficial lesions from scratching on her chest, arms, forearms, back and head. The patient also reported symptoms of anxiety, so was assessed by consultation-liaison psychiatry. Discussion: skin picking, normal behaviour in mammals, becomes pathological from a psychiatric point of view when it is repetitive and persistent, as in the case of excoriation disorder. In view of the reported relationship with the obsessive-compulsive spectrum, use of selective serotonin reuptake inhibitors and cognitive behavioural therapy are recommended.

Molecular phylogenetics of Black Cobra (Naja naja) in Pakistan

Background: Snakes are found on every continent in the world except Antarctica, and on smaller land masses. Being ecologically important, they also cause a large number of bites, leading to millions of deaths. Mitochondrial and nuclear gene sequences are being used to identify, characterize, and infer genetic biodiversity among different snake species. Furthermore, phylogenetics helps in inferring the relationships and evolutionary histories among these species. Black cobra is one of the four most venomous snakes in Pakistan. Four mitochondrial (ND4, Cytochrome b, 12S rRNA, and 16S rRNA) and four nuclear (C-mos, RAG-1, BDNF, and NT3) genes were used to trace diversity and infer the phylogenetic relationship of black cobra in Pakistan. Results: Almost similar phylogenies were obtained through maximum likelihood and Bayesian inference, showing two species of cobra in Pakistan, namely, black cobra (Naja naja) and brown cobra (Naja oxiana). All Naja species were divided into three clades: black cobra (N. naja) and brown cobra (N. oxiana) cladding with different species of Naja; N. naja (Pakistan) cladding with N. naja from Nepal; and N. oxiana showed close relationship with Naja kaouthia from Thailand and Naja siamensis from Thailand. Conclusion: It was confirmed genetically that there are two cobra species in Pakistan, i.e., black and brown cobras. This study will help in not only genetic conservation but also developing anti-venom against snake species.

Divergences between the Brazilian national information systems for recording deaths from venomous animals

This paper aims to highlight and analyze discrepancies in reporting of deaths due to venomous animals in Brazil, from 2001 to 2015, between two national information systems: The Notifiable Diseases Information System (Sistema de Informação de Agravos de Notificação - SINAN) and the Mortality Information System (Sistema de Informações sobre Mortalidade - SIM). Methods: Descriptive and comparative study of the SINAN and SIM information systems, was conducted via the following steps: collecting the death notices from SINAN and SIM; constructing tables and comparative graphics; and, only in scorpion sting fatalities, analyzing the distribution of deaths by age group as described in the specialized literature. Results: While SINAN identifies strong growth in the number of deaths from scorpion stings, SIM shows greater increase in the number of reported deaths from bee stings, especially in the South and Southeast regions. Notably, bees are the sole etiological agent that received more reports in SIM than in SINAN for every year in the period studied. The age-group distribution of the data on deaths from scorpion stings reinforced the indication of problems occurring in their registration in SINAN, especially since 2007, which may have an effect on analyses based on these data. Conclusion: Comparative analysis of these databases permits identification of important differences between profiles presented by these systems, which have equal relevance for Brazil as a whole and for its regions. These differences may influence the construction of various scenarios.(AU)

Comparison of the Electromyographic Activity in the Lower Trapezius Muscle According to Four Different Types of Exercises in Healthy Adults

PURPOSE: This study examined the most effective exercise while performing shoulder abduction below ninety degrees. METHODS: Thirty two healthy individuals (17 males, 15 females) participated and performed four exercises, 1) Posterior fly, 2) Prone row, 3) Modified prone cobra, and 4) External rotation in the prone position. Surface electromyography (sEMG) was used to measure the electrical activities for the lower, middle and upper fiber of trapezius and serratus anterior. RESULTS: A significant difference in the muscle activities of the upper/middle/lower trapezius and serratus anterior was observed among the three different positions in terms of the PF (posterior fly), PR (prone row), and MPC (modified prone cobra) (p<0.05). In post-hoc analysis, the activities of the lower and upper trapezius were significantly higher than those of the upper trapezius and serratus anterior (p<0.05). In addition, in ERP (external rotation in prone), there was a significant difference in each activity of the muscles. Post-hoc results indicated that the upper trapezius showed greater EMG activity than the other three muscles. CONCLUSION: External rotation in the prone position revealed the highest activation of the lower trapezius compared to upper trapezius muscle activity. This may be particularly useful in isolating the lower trapezius in cases where excessive scapular elevation is noted. Therefore, the most effective lower trapezius exercise should be performed below ninety degrees of shoulder abduction.

Epidemiological study of snakebite cases in Brazilian Western Amazonia

Abstract INTRODUCTION Brazil has the largest number of snakebite cases in South America, of which the large majority is concentrated in the Midwest and North. METHODS In this descriptive observational study, we assessed the epidemiological and clinical snakebite cases referred to the Centro de Medicina Tropical de Rondônia from September 2008 to September 2010. RESULTS We followed up 92 cases from admission until discharge, namely 81 (88%) men and 11 (12%) women, with a mean age of 37 years, and mainly from rural areas (91.3%). The snakebites occurred while performing work activities (63%) during the Amazon rainy season (78.3%). The vast majority of individuals presented from the Porto Velho microregion (84.7%). Approximately 95.6% of the snakebites were caused by snakes of the genus Bothrops, followed by two lachetics and two elapidics cases. Surgery was performed in 10 cases (9 fasciotomies in the lower limb and 1 amputation). No deaths were reported in this study, but 4 cases (4.3%) developed sequelae in the lower limb. CONCLUSIONS This study can contribute to a better understanding of envenomation in the state of Rondônia and thus can be useful for identifying real conditions that can increase the incidence of snakebites in this region. Moreover, the study results can serve as a basis for improving educational campaigns designed to prevent these types of snakebites, as well as for preserving snakes.

Semiología y diagnóstico diferencial de la insuficiencia cardíaca crónica / Semiology and differential diagnosis of chronic heart failure

Resumen: la insuficiencia cardiaca es un síndrome clínico de etiología multifactorial de alta prevalencia en la población mundial y latinoamericana debido al aumento en la expectativa de vida. Actualmente el diagnóstico de este se basa en los hallazgos de pruebas diagnósticas como el ecocardiograma o pépticos natriuréticos, pero es importante para el personal médico conocer e identificar de manera clara los signos y síntomas que presentan los pacientes con insuficiencia cardiaca, pues es una tendencia actual el desconocer datos semiológicos importantes que fueron descritos y refinados durante los siglos XIX y XX, y que ahora son validados por la medicina basada en la evidencia. Lo anterior cobra importancia pues el papel que debe tener la creciente tecnología no es acabar o desplazar el conocimiento clínico, si no ser una herramienta complementaria para el buen cuidado del paciente y así llegar al mejor desenlace posible. Este artículo pretende brindar datos basados en la evidencia para hacer de la clínica la parte fundamental del proceso diagnóstico y de la relación médico paciente. También intenta ser una guía útil para estudiantes de medicina o médicos que se desenvuelven en sitios con pocos recursos económicos, ya que la elección adecuada de un examen diagnóstico solo puede ser llevada a cabo mediante un buen interrogatorio y examen físico.

Abstract: heart failure is a clinical syndrome with multifactorial etiology of high prevalence in the world and Latin American population due to the increase in life expectancy. Currently the diagnosis of this is based on the findings of diagnostic tests such as echocardiogram or natriuretic peptic, but it is important for the medical staff to know and clearly identify the signs and symptoms that patients with heart failure present, as it is a current trend To ignore important semiological data that were described and refined during the nineteenth and twentieth centuries, and which are now validated by evidence-based medicine. This is important because the role of growing technology is not to end or displace clinical knowledge, if not to be a complementary tool for good patient care and thus reach the best possible outcome. This article aims to provide data based on the evidence to make the clinic the fundamental part of the diagnostic process and the patient medical relationship. It is also intended to be a useful guide for medical students or physicians operating in poorly funded areas, as the proper choice of a diagnostic examination can only be carried out through a thorough examination and physical examination.

Actiflagelin, a new sperm activator isolated from Walterinnesia aegyptia venom using phenotypic screening

Background Sperm contains a wealth of cell surface receptors and ion channels that are required for most of its basic functions such as motility and acrosome reaction. Conversely, animal venoms are enriched in bioactive compounds that primarily target those ion channels and cell surface receptors. We hypothesized, therefore, that animal venoms should be rich enough in sperm-modulating compounds for a drug discovery program. Our objective was to demonstrate this fact by using a sperm-based phenotypic screening to identify positive modulators from the venom of Walterinnesia aegyptia. Methods Herein, as proof of concept that venoms contain interesting compounds for sperm physiology, we fractionated Walterinnesia aegyptia snake venom by RP-HPLC and screened for bioactive fractions capable of accelerating mouse sperm motility (primary screening). Next, we purified each compound from the positive fraction by cation exchange and identified the bioactive peptide by secondary screening. The peptide sequence was established by Edman sequencing of the reduced/alkylated compound combined to LC-ESI-QTOF MS/MS analyses of reduced/alkylated fragment peptides following trypsin or V8 protease digestion. Results Using this two-step purification protocol combined to cell phenotypic screening, we identified a new toxin of 7329.38 Da (actiflagelin) that activates sperm motility in vitro from OF1 male mice. Actiflagelin is 63 amino acids in length and contains five disulfide bridges along the proposed pattern of disulfide connectivity C1-C5, C2-C3, C4- C6, C7-C8 and C9-C10. Modeling of its structure suggests that it belongs to the family of three finger toxins with a noticeable homology with bucandin, a peptide from Bungarus candidus venom. Conclusions This report demonstrates the feasibility of identifying profertility compounds that may be of therapeutic potential for infertility cases where motility is an issue.

True or false coral snake: is it worth the risk? A Micrurus corallinus case report

Background Bites provoked by the genus Micrurus represent less than 1% of snakebite cases notified in Brazil, a tiny fraction compared with other genus such as Bothrops and Crotalus, which together represent almost 80% of accidents. In addition to their less aggressive behavior, habits and morphology of coral snakes are determinant factors for such low incidence of accidents. Although Micrurus bites are rare, victims must be rescued and hospitalized in a short period of time, because this type of envenoming may evolve to a progressive muscle weakness and acute respiratory failure. Case Presentation We report an accident caused by Micrurus corallinus involving a 28-year-old Caucasian sailor man bitten on the hand. The accident occurred in a recreational camp because people believed the snake was not venomous. The victim presented neurological symptoms 2 h after the accident and was taken to the hospital, where he received antielapidic serum 10 h after the bite. After the antivenom treatment, the patient presented clinical evolution without complications and was discharged 4 days later. Conclusions We reinforce that it is essential to have a health care structure suitable for the treatment of snakebite. Besides, the manipulation of these animals should only be carried out by a team of well-equipped and trained professionals, and even so with special attention.

Perfil de resistencia antimicrobiana en bacilos Gram negativos no fermentadores aislados en fuentes hídricas / Antimicrobial resistance profile in non-fermenting gram negative bacilli isolated in water resources

La amplia distribución de los bacilos gram negativos no fermentadores en medios ambientales como el agua y especies vegetales cobra importancia al ser reconocidos como agentes causales de enfermedades en pacientes inmunocomprometidos, de allí la relevancia del porque debemos conocer la prevalencia y perfil de susceptibilidad de estos microorganismos en ambientes no hospitalarios. Materiales y Métodos: Estudio transversal, realizado en muestras hídricas de fuentes naturales y artificiales de almacenamiento para el consumo humano en la ciudad de Bogotá y municipios aledaños. La identificación se realizó a través de pruebas IMVIC y el perfil de resistencia a través del método de kirby bauer o E-TEST®. Resultados: Se obtuvieron 42 muestras, 7 (16,6%) con aislamientos de interés: 3 (60%) Pseudomonas spp, 2 (20%) Acinetobacter spp, 1 (10%) Sphingomonas paucimobilis y 1 (10%) Pantoea spp. El 70% presento resistencia a la ceftriaxona, el 30% a cefoxitina, 20% a gentamicina, 10% a ciprofloxacina y 10% a piperacilina-tazobactam. No se presentó resistencia a imipenem. Conclusión: 5 de 7 aislamientos revelaron un BGNNF de importancia en infección en humanos, siendo importante la resistencia encontrada a la ceftriaxona.

The wide distribution of non-fermenting gram negative bacilli in environmental media such as water and plants becomes important as they are recognized a cause of diseases in immunocompromised patients, that’s the reason why we should to know the prevalence and the susceptibility profile of these microorganisms in non-hospital environments. Materials and Methods: Cross-sectional study done with samples of natural and artificial water storage for human consumption in the city of Bogotá and surrounding municipalities. The identification was made through IMVIC tests and the resistance profile through the kirby bauer or E-TEST® method. Results: 42 samples were obtained, 7 (16.6%) with isolates of interest: 3 (60%) Pseudomonas spp, 2 (20%) Acinetobacter spp, 1 (10%) Sphingomonas paucimobilis and 1 (10%) Pantoea spp. The 70% had resistance to ceftriaxone, 30% to cefoxitin, 20% to gentamicin, 10% to ciprofloxacin and 10% to piperacillin-tazobactam. No resistance to imipenem was shown. Conclusion: 5 of 7 isolates revealed a BGNNF of importance in infection in humans, with an important resistance to ceftriaxone.

Biological and molecular properties of yellow venom of the Amazonian coral snake Micrurus surinamensis

Abstract INTRODUCTION: The coral snake Micrurus surinamensis, which is widely distributed throughout Amazonia, has a neurotoxic venom. It is important to characterize the biological and molecular properties of this venom in order to develop effective antitoxins. METHODS: Toxins from the venom of M. surinamensis were analyzed by two-dimensional polyacrylamide gel electrophoresis and their neurotoxic effects in vivo were evaluated. RESULTS AND CONCLUSIONS: Most proteins in the venom had masses < 14kDa, low phospholipase A2 activity, and no proteolytic activity. The toxins inhibited the coagulation cascade. The venom had neurotoxic effects in mice, with a median lethal dose upon intravenous administration of 700 µg/kg. Immunogenic studies revealed abundant cross-reactivity of antielapidic serum with 14kDa toxins and limited cross-reactivity with toxins < 10kDa. These results indicate that antielapidic serum against M. surinamensis venom has weak potency (0.35mg/ml) in mice.

Guangxi cobra venom-derived NGF promotes the osteogenic and therapeutic effects of porous BCP ceramic

Neuro-osteological interactions have an important role in the regulation of bone metabolism and regeneration. Neuropeptides combined with porous biphasic calcium phosphates (BCP) using protein adsorption may contribute to the acceleration of bone formation. In the present study, we investigated the effect of BCP combined with nerve growth factor (NGF) on the growth of osteoblasts in vitro and the combinational therapeutic effect on the repair of calvarial defects in vivo. NGF was separated and purified from Chinese cobra venom using a simplified three-step chromatography method. BCP combined with NGF exerted a potent effect on osteoblast differentiation, as evidenced by enhanced cell proliferation, increased ALP activity and the up-regulated expression of osteogenesis-related genes and proteins. Further, combinational therapy with BCP and NGF improved calvarial regeneration, which was superior to treatment with therapy alone, as observed using imageological and morphological examination and histological and immunohistochemical staining. The results confirmed the effect of neuro-osteological interactions through combinatorial treatment with NGF and BCP to promote osteogenesis and bone formation, which may provide an effective and economical strategy for clinical application.

Hearing loss screening tool (COBRA score) for newborns in primary care setting / 소아과

PURPOSE: To develop and evaluate a simple screening tool to assess hearing loss in newborns. A derived score was compared with the standard clinical practice tool. METHODS: This cohort study was designed to screen the hearing of newborns using transiently evoked otoacoustic emission and auditory brain stem response, and to determine the risk factors associated with hearing loss of newborns in 3 tertiary hospitals in Northern Thailand. Data were prospectively collected from November 1, 2010 to May 31, 2012. To develop the risk score, clinical-risk indicators were measured by Poisson risk regression. The regression coefficients were transformed into item scores dividing each regression-coefficient with the smallest coefficient in the model, rounding the number to its nearest integer, and adding up to a total score. RESULTS: Five clinical risk factors (Craniofacial anomaly, Ototoxicity, Birth weight, family history [Relative] of congenital sensorineural hearing loss, and Apgar score) were included in our COBRA score. The screening tool detected, by area under the receiver operating characteristic curve, more than 80% of existing hearing loss. The positive-likelihood ratio of hearing loss in patients with scores of 4, 6, and 8 were 25.21 (95% confidence interval [CI], 14.69–43.26), 58.52 (95% CI, 36.26–94.44), and 51.56 (95% CI, 33.74–78.82), respectively. This result was similar to the standard tool (The Joint Committee on Infant Hearing) of 26.72 (95% CI, 20.59–34.66). CONCLUSION: A simple screening tool of five predictors provides good prediction indices for newborn hearing loss, which may motivate parents to bring children for further appropriate testing and investigations.

Heterologous expression, protein folding and antibody recognition of a neurotoxin from the Mexican coral snake Micrurus laticorallis

Background The cysteine-rich neurotoxins from elapid venoms are primarily responsible for human and animal envenomation; however, their low concentration in the venom may hamper the production of efficient elapid antivenoms. Therefore, the aim of the present study was to produce fully active elapid neurotoxic immunogens for elapid antivenom production. Method Cysteine-rich neurotoxins showed recombinant expression in two strains of E. coli, and were purified using affinity chromatography and reverse-phase HPLC (rpHPLC). Results The cDNA of the four disulfide-bridged peptide neurotoxin Mlat1 was cloned into a modified expression vector, pQE30, which was transfected into two different E. coli strains. The recombinant toxin (HisrMlat1) was found only in inclusion bodies in M15 strain cells, and in both inclusion bodies and cytoplasm in Origami strain cells. The HisrMlat1 from inclusion bodies from M15 cells was solubilized using guanidine hydrochloride, and then purified by rpHPLC. It showed various contiguous fractions having the same molecular mass, indicating that HisrMlat1 was oxidized after cell extraction forming different misfolded disulfide bridge arrangements without biological activity. In vitro folding conditions of the misfolded HisrMlat1 generated a biologically active HisrMlat1. On the other hand, the HisrMlat1 from the cytoplasm from Origami cells was already soluble, and then purified by HPLC. It showed a single fraction with neurotoxic activity; so, no folding steps were needed. The in vitro folded HisrMlat1 from M15 cells and the cytoplasmic soluble HisrMlat1from Origami cells were indistinguishable in their structure and neurotoxicity. Rabbit polyclonal antibodies raised up against biologically active HisrMlat1 recognized the native Mlat1 (nMlat1) from the whole venom of M. laticorallis. In addition, HisrMlat1 was recognized by horse polyclonal antibodies obtained from the immunization of elapid species from sub-Saharan Africa. Conclusion HisrMlat1 shows increased biological activities compared to the native peptide, and may be used as an immunizing agent in combination with other toxic components such phospholipases type A2 for elapid antivenom production.(AU)

Assessment of the Precision and Functional Sensitivity of Two Thyroglobulin Assays: Comparison of the Second-Generation Roche Electrochemiluminescent Immunoassay and BRAHAMS Radioimmunoassay

BACKGROUND: Thyroglobulin (Tg) is the primary biochemical marker used to monitor patients with differentiated thyroid cancer (DTC) for residual or recurrent disease after total thyroidectomy, as only normal or well-differentiated malignant thyroid cells produce Tg. Here, we evaluated the precision and functional sensitivity (FS) of a recently developed highly sensitive Tg (hsTg) electrochemiluminescent immunoassay (ECLIA) and compared it to that of the radioimmunoassay (RIA) method using pooled human serum with low levels of Tg. METHODS: For the ECLIA method, the Elecsys Tg II kit (Roche Diagnostics, Germany) was used with an E170 analyzer (Roche Diagnostics). For the RIA method, the Tg-plus-RIA kit (BRAHAMS, Germany) was used with a Cobra Quantum gamma counter (Packard Instrument Company, USA). The precision and limit of detection (LOD) were determined according to the Clinical and Laboratory Standards Institute (CLSI) guidelines. FS was determined using a modification of the CLSI guideline. RESULTS: The total precision of the hsTg ECLIA and RIA methods was 9.6% and 48.2%, respectively. The manufacturer-reported LOD was verified by the hsTg ECLIA (0.04 ng/mL), but not by the RIA method (>0.08 ng/mL). The hsTg ECLIA showed better FS (0.04 ng/mL at a coefficient of variation [CV] of 10%) than the RIA method (0.37 ng/mL at a CV of 20%). CONCLUSIONS: Thus, the hsTg ECLIA performed better than the RIA method in terms of FS, which is extremely important for the early detection of residual or recurrent disease in DTC patients after total thyroidectomy. The excellent performance of the hsTg ECLIA could allow for clinical Tg measurement without thyroid-stimulating hormone stimulation, in contrast to the insufficient performance of the RIA method.