Alteraciones en la visión cromáticas por agentes neurotóxicos en pacientes de 20 a 40 años / Visual chromatic alteration by neurotoxics in patients of 20 ­ 40 age / Alteracao visual cromática por neurotóxicos em pacientes de 20 a 40 anos

El estudio busca determinar las alteraciones de la visión cromática por gentes neurotóxicos en pacientes de 20 a 40 años en la parroquia Checa ­ Ecuador y cuál de ellas genera mayor vulnerabilidad en las células nerviosas fotorreceptoras de la retina. MATERIALES Y MÉTODOS: se realizó un estudio evaluativo - descriptivo de pruebas en donde participaron 54 personas de edades entre 20 a 40 años que pertenecen al programa social bachillerato acelerado y se les aplicó la historia clínica de detección para determinar si están dentro de los criterios de inclusión. A los 54 pacientes que cumplieron con el requisito, a parte del examen visual completo, se les aplicó el test de Farnsworth que es la prueba más certera y completa para el diagnóstico de las distintas alteraciones de la visión cromática. RESULTADOS: al realizar la tabulación de datos, se observa que la alteración cromática más frecuente es la deuteranomalía, y que el agente más neurotóxico para las células fotorreceptoras retinales, es el alcohol, afectando mayormente, a la población masculina de entre 31 a 35 años, el segundo neurotóxico más agresivo, es el cigarrillo, afectando a la misma población. CONCLUSIONES: se determina que los agentes neurotóxicos inciden directamente en el daño de fotorreceptores especializados al color, el daño puede ser imperceptible al principio, pero puede afectar al paciente principalmente en el desarrollo laboral, que es cuando el paciente cae en cuenta que está presentando una alteración cromática, la disminución de la exposición a los mismos, puede revertir el daño si la frecuencia no ha sido prolongada(AU)

The study seeks to determine the alterations of color vision by neurotoxic people in patients aged 20 to 40 years in the Czech parish - Ecuador and which of them generates greater vulnerability in the photoreceptor nerve cells of the retina. MATERIALS AND METHODS: an evaluative-descriptive study of tests was carried out in which 54 people between the ages of 20 and 40 who belong to the accelerated high school social program participated and the clinical history of detection was applied to them to determine if they were within the inclusion criteria. To the 54 patients who fulfilled the requirement, apart from the complete visual examination, the Farnsworth test was applied, which is the most accurate and complete test for the diagnosis of the different color vision alterations. RESULTS: when performing the data tabulation, it is observed that the most frequent chromatic alteration is deuteranomaly, and that the most neurotoxic agent for retinal photoreceptor cells is alcohol, affecting mainly the male population between 31 and 35 years of age. The second most aggressive neurotoxic is cigarettes, affecting the same population. CONCLUSIONS: it is determined that neurotoxic agents directly affect the damage of specialized photoreceptors to color, the damage may be imperceptible at first, but it can affect the patient mainly in work development, which is when the patient realizes that he is presenting a chromatic alteration, decreasing the exposure to them, can reverse the damage if the frequency has not been prolonged.(AU)

O estudo visa determinar as alterações da visão de cores por pessoas neurotóxicas em pacientes de 20 a 40 anos na freguesia checa-Equador e qual delas gera maior vulnerabilidade nas células nervosas fotorreceptoras da retina. MATERIAIS E MÉTODOS: foi realizado um estudo descritivo-avaliativo de testes em que participaram 54 pessoas entre 20 e 40 anos que pertencem ao programa social do ensino médio acelerado e foi aplicado o histórico clínico de detecção para determinar se estavam dentro dos critérios de inclusão. Aos 54 pacientes que preencheram o requisito, além do exame visual completo, foi aplicado o teste de Farnsworth, que é o teste mais preciso e completo para o diagnóstico das diferentes alterações na visão de cores. RESULTADOS: ao realizar a tabulação dos dados, observa-se que a alteração cromática mais frequente é a deuteranomalia, e que o agente mais neurotóxico para as células fotorreceptoras da retina é o álcool, afetando principalmente a população masculina entre 31 e 35 anos. O segundo neurotóxico mais agressivo é cigarros, afetando a mesma população. CONCLUSÕES: é determinado que os agentes neurotóxicos afetam diretamente o dano dos fotorreceptores especializados à cor, o dano pode ser imperceptível a princípio, mas pode afetar o paciente principalmente no desenvolvimento do trabalho, que é quando o paciente percebe que está apresentando uma alteração cromática, diminuindo a exposição a eles pode reverter o dano se a frequência não for prolongada. (AU)

Receptores metabotrópicos de glutamato e sua relação com a Esclerose Lateral Amiotrófica (ELA) / Glutamate metabotropic receptors and its relationship with Amiotrophic Lateral Sclerosis (ALS)

A Esclerose Lateral Amiotrófica (ELA), uma doença neurodegenerativa fatal, que afeta neurônios motores superiores e inferiores, tem como fisiopatologia mais aceita a excitotoxicidade mediada por glutamato. O atual estudo tem como objetivo estabelecer a relação entre esse neurotransmissor e a ELA, a partir de uma revisão de literatura nas bases de dados Pubmed e Medline. O glutamato é o principal neurotransmissor do Sistema Nervoso Central (SNC) e a excitotoxicidade gerada pelo seu acúmulo nas fendas sinápticas é tida como um dos principais mecanismos envolvidos na fisiopatologia da ELA. Os indivíduos afetados pela ELA apresentam diminuição da expressão de determinados grupos de receptores metabotrópicos de glutamato (mGlu) nos neurônios e nas células da glia desses pacientes. Os mGlu possuem um papel de destaque na modulação da excitotoxicidade por glutamato e são subdivididos em três grupos. Os mGlus do grupo 1 amplificam as transmissões sinápticas excitatórias rápidas, e os dos grupos 2 e 3 atuam como neuroprotetores inibindo a liberação do glutamato na fenda sináptica. Os mGlus são, portanto, considerados alvos terapêuticos para a atuação de drogas que combatem a excitotoxicidade e induzem a produção de fatores neurotróficos, constituindo importante atuação no combate à ELA.

Amyotrophic Lateral Sclerosis (ALS), a fatal neurodegenerative disease that affects upper and lower motor neurons, has as the most accepted pathophysiology the glutamate-mediated excitotoxicity. The present study aims to establish the relationship between this neurotransmitter and ALS, based on a literature review in the PubMed and Medline databases. Glutamate is the main neurotransmitter of the central nervous system (CNS) and the excitotoxicity generated by its accumulation in the synaptic clefts is considered one of the main mechanisms involved in the pathophysiology of ALS. People affected by ALS present a decrease in expression of certain metabotropic glutamate receptor (mGlu) groups in neurons and glial cells of these patients. mGlu has a prominent role in modulating glutamate excitotoxicity and are subdivided into three groups. Group 1 mGlu amplifies rapid excitatory synaptic transmissions, while groups 2 and 3 act as neuroprotective agents, since among other functions they inhibit glutamate release into the synaptic cleft. Finally, mGlu are considered therapeutic targets for the action of drugs that fight excitotoxicity and induce the production of neurotrophic factors, constituting an important action in the fight against ALS.

MS/MS analysis of four scorpion venoms from Colombia: a descriptive approach

Background: Scorpions are widely known for the neurotoxic effects of their venoms, which contain peptides affecting ionic channels. Although Colombia is recognized for its scorpion diversity, only a few studies are available describing the venom content. Methods: In this descriptive study, we analyzed the MS/MS sequence, electrophoretic and chromatographic profile linked to a bioinformatics analysis of the scorpions Chactas reticulatus (Chactidae), Opisthacanthus elatus (Hormuridae), Centruroides edwardsii (Buthidae) and Tityus asthenes (Buthidae) from Colombia. Results: Each scorpion showed a specific electrophoretic and chromatographic profile. The electrophoretic profiles indicate the presence of high molecular mass compounds in all venoms, with a predominance of low molecular mass compounds in the Buthidae species. Chromatographic profiles showed a similar pattern as the electrophoretic profiles. From the MS/MS analysis of the chromatographic collected fractions, we obtained internal peptide sequences corresponding to proteins reported in scorpions from the respective family of the analyzed samples. Some of these proteins correspond to neurotoxins affecting ionic channels, antimicrobial peptides and metalloproteinase-like fragments. In the venom of Tityus asthenes, the MSn analysis allowed the detection of two toxins affecting sodium channels covering 50% and 84% of the sequence respectively, showing 100% sequence similarity. Two sequences from Tityus asthenes showed sequence similarity with a phospholipase from Opisthacanthus cayaporum indicating the presence of this type of toxin in this species for the first time. One sequence matching a hypothetical secreted protein from Hottentotta judaicus was found in three of the studied venoms. We found that this protein is common in the Buthidae family whereas it has been reported in other families - such as Scorpionidae - and may be part of the evolutionary puzzle of venoms in these arachnids. Conclusion: Buthidae venoms from Colombia can be considered an important source of peptides similar to toxins affecting ionic channels. An interesting predicted antimicrobial peptide was detected in three of the analyzed venoms.(AU)

Exploring the five-paced viper (Deinagkistrodon acutus) venom proteome by integrating a combinatorial peptide ligand library approach with shotgun LC-MS/MS

Snake venoms are complex mixtures of toxic proteins or peptides encoded by various gene families that function synergistically to incapacitate prey. In the present study, in order to unravel the proteomic repertoire of Deinagkistrodon acutus venom, some trace abundance components were analyzed. Methods Shotgun proteomic approach combined with shotgun nano-LC-ESI-MS/MS were employed to characterize the medically important D. acutus venom, after collected samples were enriched with the combinatorial peptide ligand library (CPLL). Results This avenue helped us find some trace components, undetected before, in D. acutus venom. The results indicated that D. acutus venom comprised 84 distinct proteins from 10 toxin families and 12 other proteins. These results are more than twice the number of venom components obtained from previous studies, which were only 29 distinct proteins obtained through RP-HPLC for the venom of the same species. The present results indicated that in D. acutus venom, the most abundant components (66.9%) included metalloproteinases, serine proteinases, and C-type lectin proteins; the medium abundant components (13%) comprised phospholipases A2 (PLA2) and 5'-nucleotidases and nucleases; whereas least abundant components (6%) were aminopeptidases, L-amino acid oxidases (LAAO), neurotoxins and disintegrins; and the trace components. The last were undetected before the use of conventional shotgun proteomics combined with shotgun nano-LC-ESI-MS/MS, such as cysteine-rich secretory proteins Da-CRPa, phospholipases B-like 1, phospholipases B (PLB), nerve growth factors (NGF), glutaminyl-peptide cyclortransferases (QC), and vascular non-inflammatory molecules 2 (VNN2). Conclusion These findings demonstrated that the CPLL enrichment method worked well in finding the trace toxin proteins in D. acutus venom, in contrast with the previous venomic characterization of D. acutus by conventional LC-MS/MS. In conclusion, this approach combined with the CPLL enrichment was effective for allowing us to explore the hidden D. acutus venomic profile and extended the list of potential venom toxins.(AU)

Proteomic analysis of Red Sea Conus taeniatus venom reveals potential biological applications

Diverse and unique bioactive neurotoxins known as conopeptides or conotoxins are produced by venomous marine cone snails. Currently, these small and stable molecules are of great importance as research tools and platforms for discovering new drugs and therapeutics. Therefore, the characterization of Conus venom is of great significance, especially for poorly studied species. Methods: In this study, we used bioanalytical techniques to determine the venom profile and emphasize the functional composition of conopeptides in Conus taeniatus, a neglected worm-hunting cone snail. Results: The proteomic analysis revealed that 84.0% of the venom proteins were between 500 and 4,000 Da, and 16.0% were > 4,000 Da. In C. taeniatus venom, 234 peptide fragments were identified and classified as conotoxin precursors or non-conotoxin proteins. In this process, 153 conotoxin precursors were identified and matched to 23 conotoxin precursors and hormone superfamilies. Notably, the four conotoxin superfamilies T (22.87%), O1 (17.65%), M (13.1%) and O2 (9.8%) were the most abundant peptides in C. taeniatus venom, accounting for 63.40% of the total conotoxin diversity. On the other hand, 48 non-conotoxin proteins were identified in the venom of C. taeniatus. Moreover, several possibly biologically active peptide matches were identified, and putative applications of the peptides were assigned. Conclusion: Our study showed that the composition of the C. taeniatus-derived proteome is comparable to that of other Conus species and contains an effective mix of toxins, ionic channel inhibitors and antimicrobials. Additionally, it provides a guidepost for identifying novel conopeptides from the venom of C. taeniatus and discovering conopeptides of potential pharmaceutical importance.(AU)

Analgesic effects of Phα1ß toxin: a review of mechanisms of action involving pain pathways

Phα1ß is a neurotoxin purified from spider venom that acts as a high-voltage-activated (HVA) calcium channel blocker. This spider peptide has shown a high selectivity for N-type HVA calcium channels (NVACC) and an analgesic effect in several animal models of pain. Its activity was associated with a reduction in calcium transients, glutamate release, and reactive oxygen species production from the spinal cord tissue and dorsal ganglia root (DRG) in rats and mice. It has been reported that intrathecal (i.t.) administration of Phα1ß to treat chronic pain reverted opioid tolerance with a safer profile than ω-conotoxin MVIIA, a highly selective NVACC blocker. Following a recent development of recombinant Phα1ß (CTK 01512-2), a new molecular target, TRPA1, the structural arrangement of disulphide bridges, and an effect on glial plasticity have been identified. CTK 01512-2 reproduced the antinociceptive effects of the native toxin not only after the intrathecal but also after the intravenous administration. Herein, we review the Phα1ß antinociceptive activity in the most relevant pain models and its mechanisms of action, highlighting the impact of CTK 01512-2 synthesis and its potential for multimodal analgesia.

Analgesic effects of Phalfa1 beta toxin: a review of mechanisms of action involving pain pathways

Phα1ß is a neurotoxin purified from spider venom that acts as a high-voltage-activated (HVA) calcium channel blocker. This spider peptide has shown a high selectivity for N-type HVA calcium channels (NVACC) and an analgesic effect in several animal models of pain. Its activity was associated with a reduction in calcium transients, glutamate release, and reactive oxygen species production from the spinal cord tissue and dorsal ganglia root (DRG) in rats and mice. It has been reported that intrathecal (i.t.) administration of Phα1ß to treat chronic pain reverted opioid tolerance with a safer profile than ω-conotoxin MVIIA, a highly selective NVACC blocker. Following a recent development of recombinant Phα1ß (CTK 01512-2), a new molecular target, TRPA1, the structural arrangement of disulphide bridges, and an effect on glial plasticity have been identified. CTK 01512-2 reproduced the antinociceptive effects of the native toxin not only after the intrathecal but also after the intravenous administration. Herein, we review the Phα1ß antinociceptive activity in the most relevant pain models and its mechanisms of action, highlighting the impact of CTK 01512-2 synthesis and its potential for multimodal analgesia.

Transtorno do espectro autista: uma revisão sistemática sobre aspectos nutricionais e eixo intestino-cérebro / Autism spectrum disorder: a systematic review about nutritional aspects and gut-brain axis

O número de indivíduos diagnosticados com o transtorno do espectro autista (TEA) registrou aumento evidente na última década. Os principais sintomas, apresentados pelo portador, são neurológicos e digestórios, estando às intervenções nutricionais dentre as terapêuticas mais promissoras para amenizar a sintomatologia clínica. Assim, objetivou-se revisar sistematicamente os estudos sobre distúrbios alimentares e do trato gastrointestinal apresentado pelo indivíduo portador do TEA, a fim de compreender como o comportamento alimentar influência na etiopatogênese e manifestações clínicas da doença, com foco no eixo intestinocérebro. Para isso realizou-se uma revisão sistemática, seguindo as diretrizes PRISMA. A partir de uma busca estruturada e abrangente em bases de dados eletrônicas, 23 estudos foram recuperados e incluídos na revisão. Os critérios de inclusão definiam ser artigos originais relacionando o TEA com alterações nutricionais e/ou com o eixo intestino-cérebro. Após análise da composição da microbiota intestinal, os estudos mostraram um quadro de desequilíbrio. Foram encontradas, também, alterações na barreira de muco e permeabilidade intestinal e alterações em proteínas envolvidas na digestão e absorção de alimentos. Dietas restritivas e a modulação da microbiota, com uso de probióticos e de antibióticos específicos, são apresentadas como estratégias terapêuticas adjuvantes promissoras. Conclui-se que o eixo intestino-cérebro está envolvido tanto na etiologia, quanto nas manifestações clínicas do TEA. Porém, não sendo certo se alterações intestinais são causa ou consequência das alterações neurológicas. Até o presente momento, a comunidade científica não tem conclusões suficientes para indicar o uso de dietas restritivas, e uso de probióticos e de antibióticos como terapêutica para o TEA.

The number of individuals diagnosed with autism spectrum disorder (ASD) had an evident increase in the last decade. The primary symptoms exhibited amongst these patients were mostly digestive and neurological disorders; with nutritional interventions being one of the most promising therapies to assuage this clinical symptomology. As such, following the PRISMA guidelines, we systematically reviewed the research studies apropos of the ASD patients manifesting said digestive disorders, to comprehend how dietary behavior can influence the etiopathogenesis and clinical manifestations of the disease, with primary focus on the gut-brain axis. From a comprehensive and structured search through electronic databases, 23 studies were retrieved and admitted in this review. The inclusion criteria defined that there be original articles consociating ASD with nutritional disorders and/or with the gut-brain axis. These studies analyzed the composition of the intestinal flora of diagnosed patients, subsequently discerning cases of varying imbalances. Alterations in the gene expression of the proteins involved in the digestion and absorption of food, the mucous barrier and the intestinal permeability were described. Accordingly, restrictive diets and the modulation of the microbiota by administering specific anti- & probiotics were initially identified as promissory therapies. In conclusion, the gut-brain axis was observed to be a determinant factor in both the etiology and clinical symptomology of ASD - though it is still debatable the correlation of intestinal alterations with neurological changes. At present, there is no concrete scientific proof accrediting to restrictive diets and the use of specific anti- & probiotics, as successful treatments for ASD.

Epidemiologia e fatores de riscos relacionados à intoxicação alimentar causada por Clostridium Botulinum: uma revisão narrativa / Epidemiology and risk factors related to food intoxication caused by Clostridium Botulinum: a narrative review

O botulismo é uma doença resultante da ação de uma toxina produzida pelo Clostridium botulinum. Devido à sua gravidade e alta mortalidade é considerado um problema de saúde pública. Nesta revisão apresentamos os principais fatores de riscos associados à intoxicação alimentar provocada pelo Clostridium botulinum, bem como realizamos um levantamento epidemiológico sobre o botulismo alimentar e infantil. A busca bibliográfica considerou as bases de dados Scielo, Medline, Lilacs e PubMed. Foram selecionados artigos originais e relatos de caso publicados em inglês, espanhol e português, incluindo publicações dos últimos dez anos. A partir das análises dos títulos, resumos e artigos, um total de 26 artigos foram incluídos nesta revisão. Verificou-se predomínio de 54% dos casos referentes ao botulismo alimentar, dos quais aproximadamente 58% confirmaram a ocorrência da toxina tipo A; e 35% referente ao botulismo infantil. Na literatura consultada os principais sintomas, relacionados ao botulismo alimentar, identificados foram: visão turva, vômito, paralisia flácida, náuseas, tontura, diplopia, dificuldade respiratória, disatria, disfagia, fraqueza muscular, boca seca, ptose e cefaleia. Dentre as principais fontes de contaminação, 65% das publicações selecionadas identificaram as conservas como principal causa do botulismo alimentar. Embora o mel (42%) seja a única fonte registrada de alimento veiculador do agente causador do botulismo infantil, alguns relatos na literatura (25%) associaram à doença com a inalação de poeira contendo esporos do Clostridium botulinum, bem como o uso de plantas medicinais (25%). Os sintomas mais comuns observados na literatura foram: constipação dificuldade respiratória e dificuldade de sucção. Apesar de vários relatos na literatura acerca das duas doenças, o botulismo ainda é muito subnotificado dado ao diagnóstico muitas vezes equivocado, ressaltando-se a importância do diagnóstico precoce no tratamento da doença pelos profissionais de saúde, bem como a disponibilidade de informações relevantes para a investigação epidemiológica de doenças de notificação compulsória. Os dados apresentados também demonstram a importância de sensibilizar a população dos principais riscos e medidas de prevenção, já que a maioria dos casos relatados está relacionada a práticas inadequadas de preparo dos alimentos. (AU)

Botulism is a disease resulting from the action of a toxin produced by Clostridium botulinum. Because of its severity and high mortality, it is considered a public health problem. In this review, we present the main risk factors associated with food poisoning caused by Clostridium botulinum, as well as an epidemiological survey on foodborne and infant botulism. A bibliographic search was conducted in SciELO, MEDLINE, LILACS and PubMed databases. Original articles and case reports published in English, Spanish and Portuguese in the past ten years were selected. After analyzing titles, abstracts and articles, 26 articles were used in this review. In total, 54% of the cases were related to foodborne botulism, of which approximately 58% had confirmed type A botulism, and 35% were related to infant botulism. In the literature consulted, the main symptoms related to foodborne botulism were blurred vision, vomiting, flaccid paralysis, nausea, dizziness, diplopia, respiratory distress, dysarthria, dysphagia, muscle weakness, dry mouth, ptosis and headache. Among the sources of contamination, 65% of the published studies reported home-canned foods as the main cause of foodborne botulism. Although honey (42%) is the only reported food source for the agent causing infant botulism, some reports in the literature (25%) associated the disease with inhalation of dust containing Clostridium botulinum spores, as well as use of medicinal plants (25%). The most common symptoms observed in the literature were constipation, difficulty breathing and difficulty suckling. Although several reports on the two forms of the disease exist, botulism remains under-reported because of often incorrect diagnosis. Thus, early diagnosis is important for an adequate treatment provided by health professionals, as well as availability of relevant information for the epidemiological investigation of notifiable diseases. The data presented in this study also demonstrate the importance of raising people's awareness to main risks and prevention measures, as most reported cases were related to inadequate food preparation practices. (AU)

Proteome of fraction from Tityus serrulatus venom reveals new enzymes and toxins

Tityus serrulatus venom (Ts venom) is a complex mixture of several compounds with biotechnological and therapeutical potentials, which highlights the importance of the identification and characterization of these components. Although a considerable number of studies have been dedicated to the characterization of this complex cocktail, there is still a limitation of knowledge concerning its venom composition. Most of Ts venom studies aim to isolate and characterize their neurotoxins, which are small, basic proteins and are eluted with high buffer concentrations on cation exchange chromatography. The first and largest fraction from carboxymethyl cellulose-52 (CMC-52) chromatography of Ts venom, named fraction I (Fr I), is a mixture of proteins of high and low molecular masses, which do not interact with the cation exchange resin, being therefore a probable source of components still unknown of this venom. Thus, the present study aimed to perform the proteome study of Fraction I from Ts venom, by high resolution mass spectrometry, and its biochemical characterization, by the determination of several enzymatic activities. Methods: Fraction I was obtained by a cation exchange chromatography using 50 mg of crude venom. This fraction was subjected to a biochemical characterization, including determination of L-amino acid oxidase, phospholipase, hyaluronidase, proteases activities and inhibition of angiotensin converting enzyme (ACE) activity. Fraction I was submitted to reduction, alkylation and digestion processes, and the tryptic digested peptides obtained were analyzed in a Q-Exactive Orbitrap mass spectrometer. Data analysis was performed by PEAKS 8.5 software against NCBI database. Results: Fraction I exhibits proteolytic activity and it was able to inhibit ACE activity. Its proteome analysis identified 8 different classes of venom components, among them: neurotoxins (48%), metalloproteinases (21%), hypotensive peptides (11%), cysteine-rich venom protein (9%), antimicrobial peptides (AMP), phospholipases and other enzymes (chymotrypsin and lysozymes) (3%) and phosphodiesterases (2%). Conclusions: The combination of a proteomic and biochemical characterization strategies leads us to identify new components in the T. serrulatus scorpion venom. The proteome of venom´s fraction can provide valuable direction in the obtainment of components in their native forms in order to perform a preliminary characterization and, consequently, to promote advances in biological discoveries in toxinology.(AU)

Diseño de un programa de análisis de peligros y puntos críticos control (HACCP) para la producción de toxoide tetánico / Design of a program of hazard analysis and critical control point (HACCP) for production of Tetanus Toxoid

El toxoide tetánico es una neurotoxina modificada que induce la formación de una antitoxina protectora contra la enfermedad denominada tétanos. Este antígeno es obtenido a partir de procesos fermentativos con la bacteria anaerobia Clostridium tetani y es utilizado para la formulación de vacunas simples y combinadas inactivadas. Con el propósito de atender a las recomendaciones y regulaciones de la Organización Mundial de la Salud (OMS), el objetivo de este trabajo fue diseñar un Programa de Análisis de Peligros y Puntos Críticos de Control (HACCP) en la producción del antígeno Toxoide Tetánico, desde la recepción de la cepa certificada en el área de producción hasta el almacenamiento del toxoide tetánico purificado. Para ello, inicialmente se evaluó el cumplimiento de los prerequisitos (BPM, POES, BPL). Posteriormente, se procedió al diseño del plan HACCP mediante la ejecución de las 5 tareas preliminares y la aplicación de los 7 principios, conforme a la metodología descrita por la OMS. A partir del análisis de peligros en todas las etapas del proceso de producción del toxoide tetánico se identificaron 3 puntos críticos de control: detoxificación, filtración estéril final y almacenamiento de toxoide tetánico purificado. Se establecieron los límites críticos, los procedimientos de vigilancia, las acciones correctivas, los procedimientos de verificación y de documentación. La propuesta tiene como fin garantizar la calidad e inocuidad del producto elaborado, la protección del personal involucrado en el proceso y del medio ambiente con miras a la obtención de la certificación como laboratorio productor de vacunas

Tetanus toxoid is a modified neurotoxin that induces the formation of protective antitoxin of the disease called tetanus. This antigen is obtained from fermentation processes with anaerobic bacteria Clostridium tetani and it is used to formulate simple and combined inactivated vaccines. In order to meet the recommendations and regulations of World Health Organization (WHO), the aim of this work was to design a Program of Hazard Analysis and Critical Control Points (HACCP) in the production of antigen Tetanus Toxoid, starting from the receipt of the certified strain in the production area through the storage of purified tetanus toxoid. For this, initially fulfilling the prerequisites (GMP, SSOP and GLP) was evaluated. Subsequently, we proceeded to design HACCP plan by running 5 preliminary tasks and application of the 7 principles, according to the methodology described by WHO. From the hazard analysis at all stages of the production process of tetanus toxoid three critical control points were identified: detoxification, final sterile filtration and storage of purified tetanus toxoid. Critical limits, monitoring procedures, corrective actions, verification and documentation procedures were established. The proposal aims to assure the quality and safety of the final product, the protection of personnel involved in the process and the environment, with a view to obtaining certification as a vaccine production laboratory

Surto de botulismo tipo C em equinos no Rio Grande do Sul / Outbreak of type C botulism in horses in Rio Grande do Sul

O botulismo nos animais domésticos é uma intoxicação fatal ocasionada pela ingestão da neurotoxina de Clostridium botulinum. Este trabalho descreve os aspectos epidemiológicos, clínicos e laboratoriais de um surto de botulismo tipo C em equinos de uma propriedade rural no município de Porto Alegre, Rio Grande do Sul. Do total de 20 equinos, sete (35%) foram acometidos e morreram, e um eutanasiado após apresentar prognóstico desfavorável. Os cavalos, criados extensivamente, apresentaram sinais clínicos semelhantes, caracterizados por dificuldade de locomoção, letargia, paralisia flácida progressiva, tremores musculares, sudorese, ataxia e quedas frequentes. Quatro equinos foram necropsiados e não apresentaram lesões macroscópicas e microscópicas significativas. Amostras de encéfalo e medula espinhal foram enviados para realização do exame de imunofluorescência direta e foram negativas para o vírus da raiva. Fragmentos de fígado dos quatro equinos necropsiados, amostras de sedimento do solo e água suspeita de dois açudes da propriedade foram avaliados pelo exame direto para a verificação da presença de toxina botulínica e/ou esporos de Clostridium botulinum. No bioensaio em camundongos e soroneutralização pode-se detectar toxina botulínica tipo C em uma amostra de fígado, ao passo que as amostras de sedimento do solo e água foram negativas em todos os ensaios. Com base nos dados epidemiológicos, clínicos e laboratoriais pode-se estabelecer o diagnóstico etiológico de botulismo causado pela toxina botulínica do tipo C, e sugere-se, a água estagnada dos açudes como aprovável fonte da toxina.(AU)

Botulism in domestic animals is a fatal poisoning caused by ingestion of Clostridium botulinum neurotoxin. This paper describes epidemiological, clinical and laboratorial aspects of a botulism type C outbreak in horses in a farm in Porto Alegre, Rio Grande do Sul. From a total of 20 horses, seven (35%) were affected and died, and one was euthanized after presenting poor prognosis. The horses, which were raised extensively, showed similar clinical signs, that included limited mobility, lethargy, progressive flaccid paralysis, muscle tremors, sweating, ataxia, and frequent falls. Four horses were necropsied and did not show significant macroscopic and microscopic lesions. Brain and spinal cord samples were analyzed by direct immunofluorescence and were negative for rabies. Liver fragments from the four necropsied horses, soil sediment samples, and suspect water from two reservoirs were evaluated by direct exam to verify the botulinum toxin and/or Clostridium botulinum spores presence. In the serum neutralization bioassay in mice, the type C botulinum toxin was detected in one liver sample, at the same time that the soil sediment and water samples were negative in all assays. Based on the epidemiological, clinical and laboratory data the etiological diagnosis of botulism was established by being caused by botulinum toxin type C, and it is suggested that the stagnant water of the dams is the likely source of the toxin.(AU)

Botulinum toxin for treating unilateral apraxia of eyelid opening in a patient with congenital myotonia / Tratamento da apraxia de abertura palpebral unilateral em portador de miotonia congênita com botulinum toxin

ABSTRACT A 37-year-old female presented with severe apraxia of lid opening (ALO) affecting the right upper lid associated with Becker congenital myotonia (MC). The patient had a history of right upper lid ptosis for 25 years that was exacerbated over the previous month with severe incapacity to open her right eye. No other associated neurological or ophthalmic symptoms were observed. The patient was treated with botulinum toxin (BoNT-A) injection into the pretarsal and lateral canthus region of the orbicularis oculi of the affected eyelid. Treatment with BoNT-A is an effective method of managing ALO in Becker MC. This is the first case of unilateral ALO in the course of Becker MC that was successfully treated with injections of botulinum toxin.

RESUMO Trata-se de uma mulher de 37 anos apresentando grave apraxia de abertura da pálpebra (AAP) superior direita associada com miotomia congênita de Becker (MC). A paciente há 25 anos apresentava ptose palpebral a direita e há um mês desenvolveu incapacidade de abertura do olho direito. Não havia associação com outro sintoma neurológico ou oftalmológico. A paciente recebeu injeção de botulinum toxin (BoNT-A) no músculo orbicular a direita, na região pretarsal e no canto lateral. A BoNT-A foi efetiva para o tratamento da AAP associada com miotomia congênita de Becker.

La toxina botulínica como adyuvante en el tratamiento de la sonrisa gingival / Botulinum toxin as an adjuvant in the treatment of gummy smile

RESUMEN: La búsqueda de la excelencia estética es un objetivo importante de la odontología. La sonrisa gingival es una de las principales quejas estéticas, ya que perjudica la autoestima y las relaciones sociales del paciente. Nuevas técnicas como la aplicación de la toxina botulínica (TB) pueden tornarse una opción terapéutica más conservadora, y ayudar a disminuir las proporciones de las intervenciones quirúrgicas resectivas. El propósito de este trabajo es describir la aplicación de la TB como adyuvante de la cirugía gingival resectiva, a través del reporte de un caso clínico de discrepancia dentogingival y sonrisa gingival. Con la cirugía resectiva el equilibrio dentogingival fue mejorado, y la aplicación de TB tipo A causó la dehiscencia uniforme del labio superior, devolviendo la armonía facial. La TB es un complemento útil y conservador en la mejora estética de la sonrisa, y puede aumentar los resultados de la cirugía gingival resectiva.

ABSTRACT: The search for aesthetic excellence is an important goal in dentistry. The gummy smile is one of the main aesthetic complaints as it can affect self-esteem, and prejudice the patients' social relationships. New techniques, such as the application of botulinum toxin (BT) may become a more conservative treatment option and help to reduce the proportions of resective surgery. The purpose of this paper is to describe the application of BT, used as an adjuvant to gingival resection surgery, using a case report of a dento-gingival discrepancy and gummy smile. The resection surgery improved the dento-gingival equilibrium and the application of BT led to a uniform dehiscence of the upper lip, and facial harmony. BT is a useful and conservative adjuvant in the aesthetic enhancement of the smile, and can improve the outcomes of gingival resection surgery.

Crotalus durissus terrificus crotapotin naturally displays preferred positions for amino acid substitutions

Background: Classically, Crotalus durissus terrificus (Cdt) venom can be described, according to chromatographic criteria, as a simple venom, composed of four major toxins, namely: gyroxin, crotamine, crotoxin and convulxin. Crotoxin is a non-covalent heterodimeric neurotoxin constituted of two subunits: an active phospholipase A2 and a chaperone protein, termed crotapotin. This molecule is composed of three peptide chains connected by seven disulfide bridges. Naturally occurring variants/isoforms of either crotoxin or crotapotin itself have already been reported. Methods: The crude Cdt venom was separated by using RP-HPLC and the toxins were identified by mass spectrometry (MS). Crotapotin was purified, reduced and alkylated in order to separate the peptide chains that were further analyzed by mass spectrometry and de novo peptide sequencing. Results: The RP-HPLC profile of the isolated crotapotin chains already indicated that the α chain would present isoforms, which was corroborated by the MS and tandem mass spectrometry analyses. Conclusion: It was possible to observe that the Cdt crotapotin displays a preferred amino acid substitution pattern present in the α chain, at positions 31 and 40. Moreover, substitutions could also be observed in ß and γ chains (one for each). The combinations of these four different peptides, with the already described chains, would produce ten different crotapotins, which is compatible to our previous observations for the Cdt venom.(AU)

O uso da toxina botulínica na correção do sorriso gengival- revisão de literatura / Use of the botulinum toxin in correction of the gingival smile - Literature review

A toxina botulínica inicialmente era utilizada apenas para tratamentos terapêuticos, depois de alguns estudos sua aplicação passou a ser também utilizada para tratamentos estéticos. Esta substância é produzida pela bactéria Clostridium botulinum, responsável pelo botulismo, doença que provoca intoxicação por alimentos mal conservados. Utilizada na medicina para tratar inicialmente patologias, hoje seu uso passa a ser também estético na odontologia. Alguns pacientes se queixam do sorriso gengival causado por uma exposição excessiva da gengiva e buscam tratamentos mais rápidos e com menor morbidade, neste caso a toxina botulínica é muito eficaz para a correção do sorriso gengival diagnosticado por uma hiperfunção muscular. Quando causado por excesso vertical maxilar, extrusão ou erupção passiva alterada dos dentes ântero-superiores, o sorriso gengival pode ser tratado com a cirurgia ortognática, tratamento ortodôntico, miectomia, reposicionamento labial e/ou gengivoplastia. O uso desta substância na odontologia, pode ser mais uma opção para amenizar ou corrigir o sorriso gengival por hiperfunção muscular ou associar a outros tipos de tratamentos. Desta forma, o objetivo deste trabalho é fazer uma revisão literária sobre o uso da toxina botulínica na correção do sorriso gengival e descrever seu protocolo de uso (AU)

Botulinum toxin initially was used only for therapeutic treatments, after some studies its application was also used for aesthetic treatments. This substance was produced by the bacterium Clostridium botulinum, responsible for botulism, a disease that causes intoxication by poorly preserved foods. Used in medicine to treat initially pathologies, today its use is now also aesthetic in dentistry. Some patients complain of the gingival smile caused by excessive exposure of the gums and seek faster treatments with lower morbidity; in this case the botulinum toxin is very effective for the correction of the gingival smile diagnosed by a muscular hyperfunction. When caused by excessive vertical jaw, extrusion or passive eruption of the upper anterior teeth, the gingival smile may be treated with orthognathic surgery, orthodontic treatment, myectomy, lip repositioning and / or gingivoplasty. The use of this substance in dentistry may be another option to soften or correct the gingival smile due to muscular hyperfunction or to associate with other types of treatments. In this way, the objective of this work is to make a literary review on the use of botulinum toxin in the correction of the gingival smile and to describe its protocol of use. (AU)

Phoneutrismo en Piura: reporte de un caso clínico / Phoneutrism in Piura: a case report

Se presenta un caso de mordedura por Phoneutria nigriventer atendido en el C.S. I-1 ôEl Ingenioõ en el año 2015. El caso fue de sintomatología leve a moderada y de muy buena evolución en un paciente escolar por entrada accidental de la especie en su calzado. El phoneutrismo es un aracneismo poco habitual en el Perú debido al gran desconocimiento por parte del personal médico y paramédico con el consecuente subregistro de dichos casos; sobre todo en zonas rurales dedicadas a la producción de plátanos, observándose adaptación de la especie en la costa norte; por ello presentamos este caso, el cual permitirá identificar éstos accidentes para su apropiado manejo y control...

A case of a bite by Phoneutria nigriventer was treated in El Ingenio Health Center in 2015. The case had mild to moderate symptoms and it had a very good outcome in a schoolaged patient who had the bite because of an accidental entry of the spider in her shoes. Phoneutrism is a rare arachnidism in Peru, and medical and paramedical personnel are hardly aware of this condition, particularly in rural areas dedicated to banana production. There is adaptation of this species in the Peruvian Northern coast, and this is the reason why we present this case, so our colleagues may properly identify these cases for their proper management and control...

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)

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

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.

A comparison between the recombinant expression and chemical synthesis of a short cysteine-rich insecticidal spider peptide

Background:The choice between heterologous expression versus chemical synthesis for synthesizing short cysteine-rich insecticidal peptides from arthropods may impact the obtainment of yields and well-folded bioactive molecules for scientific research. Therefore, two recombinant expression systems were compared to that of chemical synthesis for producing Ba1, a cysteine-rich spider neurotoxin.Methods:The transcription of the insecticidal neurotoxin Ba1 was obtained from a cDNA library of venom glands of the spider Brachypelma albiceps.It was cloned into the pCR®2.1-TOPO® cloning vector and then introduced in two different expression vectors, pQE40 and pET28a+. Each vector was transfected into E. coli M15 and BL21 cells, respectively, and expressed under induction with isopropyl thiogalactoside (IPTG). The chemical synthesis of Ba1 was performed in an Applied Biosystems 433A peptide synthesizer.Results:Both expression systems pQE40 and pET28a+ expressed the His-tagged recombinant protein products, HisrDFHRBa1 and HisrBa1, respectively, as inclusion bodies. The recombinant proteins HisrDFHRBa1 and HisrBa1 presented respective molecular masses of 28,289 and 8274.6 Da, and were not biologically active. These results suggested that both HisrDFHRBa1 and HisrBa1 were oxidized after cell extraction, and that their insecticidal activities were affected by their N-terminal pro-peptides and different disulfide bridge arrangements. The respective protein expression yields for HisrDFHRBa1 and HisrBa1 were 100 μg/L and 900 μg/L of culture medium. HisrBa1 was reduced and folded under in vitroconditions. The in vitro folding of HisrBa1 produced several isoforms, one of which, after removing its N-terminal pro-peptide by enzymatic cleavage, presented elevated insecticidal activities compared to the native Ba1. Furthermore, the His-tagged protein HisrDFHRBa1 underwent enzymatic cleavage to obtain recombinant Ba1 (rBa1). As expected, the molecular mass of rBa1 was 4406.4 Da. On the other hand, Ba1 was chemically synthesized (sBa1) with a yield of 11 mg per 0.1 mmol of amino acid assembly.Conclusions:The two recombinant insecticidal peptides and the one synthesized chemically were as active as the native Ba1; however, toxin yields differed drastically.(AU)