Determination of cyanide in whole seeds and brans of linseed (Linum usitatissimum Linn) by molecular spectrophotometry

Abstract The addition of linseed (Linum usitatissimum Linn) in the diet, as a functional food, has increased over the years. However, it possesses cyanogenic glycosides. This study aimed to quantify and compare cyanide concentration in whole seed and bran of brown and golden types to establish a safe limit of intake. Three commercial labels, from brown and golden whole seed types (Ab, Ag, Bb, Bg, Cb and Cg), and six commercial labels of brown and golden bran (1b, 2g, 3g, 4b, 5g, and 6b), were selected, totalizing twelve samples. Total cyanide concentration was quantified by a colorimetric method employing alkaline picrate, after acid hydrolysis. The whole seed cyanide values were between 348.4 and 473.20 µg/g and the bran cyanide values were between 459.53 and 639.35 µg/g. The analyzed bran presented increased cyanide concentrations than the whole seeds with no differences between brown and golden types. Food able to produce cyanide less than 90 µg/kg body weight, daily, is considered secure for consumption. Considering this limit and analyzed samples, it is safe to eat approximately two tablespoons of seeds or one tablespoon of bran. These results point out the importance of cyanide amount daily intake information to be in linseed packaging, to ensure secure consumption

Studies on the enzymatic specificity of diguanylate cyclases domains and the production of new second messengers / Estudos na especificidade enzimática de diguanilato ciclases e na produção de novos segundos mensageiros

In the first chapter, studies on substrate recognition and enzymatic activity of GGDEF domains are presented. Many proteins containing GGDEF domains are diguanylate cyclases (DGCs, EC 2.7.7.65), enzymes that catalyze the conversion of 2 GTP molecules into the second messenger c-di-GMP in prokaryotes. This molecule is primarily implicated in the transition between motile and sessile lifestyles, as well several other phenotypes. Redundancy and diversity of GGDEF domain sequences in many bacterial genomes raises the possibility that other enzymatic functions may yet be discovered. To test this hypothesis, i) the effect of point mutations on the structure and enzymatic activity of GGDEF domains is analyzed, ii) the enzymatic specificity of wild-type GGDEF domains from different proteins is also tested, and iii) when non-canonical products are detected, enzymatic models are studied to understand its preferential production. The principal results obtained from these studies are as follows. Seven mutants of the DGC PleD (a GGDEF containing-protein from Caulobacter crescentus) were constructed and the crystallographic structure of two of them was solved, showing that they are unlikely to bind the guanine moiety in its active site. Additionally, five mutants of XAC0610, another DGC from Xanthomonas citr, were constructed and their substrate specificities were evaluated. None of those mutants were able to use ATP as a substrate. Finally, seven different GGDEF domain-containing DGCs from different sources were expressed and purified and their enzymatic specificities were tested with several nucleotide triphosphates. One enzyme, GSU1658 from Geobacter sulfurreducens was particularly promiscuous and shown to produce c-di-GMP, c-di-AMP, c-di-IMP, c-di-2´dGMP, cGAMP, c-GIMP, and c-AIMP. Interestingly, XAC0610 was able to recognize 2´dGTP as substrate. Analysis of enzyme kinetics of XAC0610 in presence of 2´dGTP and/or GTP showed the preferential formation of the hybrid linear product pppGp2´dG. The second chapter present studies on cyanide metabolism in Bacillus with focus on the cyanide dihydratase of Bacillus safensis. Cyanide is widely used in industries due to its high affinity for metals. This same ability confers potent toxicity to this compound. Thus, industries must reduce the cyanide concentration from wastewater before its final disposal. Physical, chemical, and biological methods have been developed to achieve this goal, but knowledge about metabolic pathways and the biology of enzymes involved in cyanide degradation is still scarce. Here, the isolation of a Bacillus safensis strain from mine tailings in Peru is described. Classification of this strain was done through a comparative analysis of 132 core genomes of strains from the Bacillus pumilus group. Sequence analysis determined that a cyanide dihydratase (CynD, EC 3.5.5.1)) encoded in the genome of the isolated strain was likely the enzyme responsible for cyanide degradation. Confirmation of the cyanide degrading activity of CynD from this strain was achieved by cloning, expression and purification of the enzyme and its enzymatic characterization. CynD from this strain was active up to pH 9 and oligomerization patterns analyzed by SEC-MALS and electron microscopy showed that the enzyme forms large helical structures at pH 8 and smaller structures at higher pHs. Finally, we show that CynD expression is strongly induced in the presence of cyanide. The last two years of graduate studies were carried out in the context of the COVID-19 pandemic. Thanks to the large amount of publicly available genomic data, we were able to carry out studies on the worldwide dynamics of the spread of SARS-CoV-2 mutants forms. In the first year of the pandemic, genomic classification of 171,461 genomes showed the presence of five major haplotypes based on nine mutations. The worldwide distribution and the temporal evolution of frequency of these haplotypes was carefully analyzed. All the haplotypes were identified in the six regions analyzed (South America, North America, Europe, Asia, Africa, and Oceania); however, the frequency of each of them was different in each of these regions. As of September 30, 2020, haplotype 3 (or operational taxonomic unit 3, OTU_3) was the most prevalent in four regions (South America, Asia, Africa, and Oceania). OTU_5 was the most prevalent in North America and OTU_2 in Europe. Temporal dynamics of the haplotypes showed that OTU_1 became nearly extinct after 8 months of pandemic (November 2020). Other OTUs are still present in different frequencies all around the world, while currently generating new variants. Based on their temporal dynamics, a classification scheme of 115 SARS-CoV-2 mutations identified from 1,058,020 SARS-COV-2 genomes was also performed. Three types of temporal dynamics of mutations were identified: i) High-Frequency mutations are characterized by a rapid increase in frequency upon its appearance, ii) medium and iii) low-frequency mutations maintain mid or low-frequencies for several months and can be region-specific. Finally, we performed a correlation analysis of the effective reproduction number (Rt) of SARS-CoV-2 harboring the high-frequency mutation N501Y with the level of control measures adopted in specific jurisdictions. We show that Rt is negatively correlated with the level of control measures in eight of the nine countries analyzed. This negative correlation was similar when we analyzed the Rt of SARS-CoV-2 not-harboring N501Y. Thus, the control measures likely diminish the Rt of both SARSCoV-2 wild-type and N501Y

O presente trabalho está dividido em três capítulos sobre linhas de pesquisa diferentes desenvolvidas pelo autor durante o período de doutorado No primeiro capítulo, são apresentados estudos relacionados ao reconhecimento estrutural de substratos e análise enzimática de domínios GGDEF com atividade diguanilato ciclase (EC 2.7.7.65). As proteínas contendo domínios GGDEF estão relacionados à produção enzimática do segundo mensageiro c-di-GMP, a partir de duas moléculas de GTP, em procariotos. Esta molécula está principalmente envolvida na transição entre os estilos de vida móveis e sésseis, bem como vários outros fenótipos. Redundância e diversidade de sequências de domínio GGDEF aumentam a possibilidade de que outras funções enzimáticas ainda possam ser descobertas. Para testar esta hipótese, i) o efeito de mutações pontuais na estrutura e atividade enzimática dos domínios GGDEF é analisado, ii) a especificidade enzimática de domínios GGDEF de enzimas diferentes também é testada e iii) quando produtos não canônicos são detectados, modelos enzimáticos são estudados para entender sua produção preferencial. Como resultados mais importantes, sete mutantes do PleD (uma proteína contendo GGDEF) foram construídos e a estrutura cristalográfica de dois delas foi resolvida, mostrando que é improvável que eles liguem à porção guanina em seu sítio ativo. Além disso, cinco mutantes da proteína XAC0610 de Xanthomonas citri foram construídos e sua capacidade de usar ATP ou GTP como substrato foi avaliada. Nenhum desses mutantes foi capaz de usar ATP como substrato. Finalmente, sete outras proteínas contendo GGDEF foram purificadas e sua especificidade enzimática foi avaliada com vários trifosfatos de nucleotídeos. Uma enzima promíscua chamada GSU1658 mostrou produzir c-di-GMP, c-di-AMP, c-di-IMP, c-di-2´dGMP, c-GAMP, cGIMP e c-AIMP. Curiosamente, o XAC0610 foi capaz de reconhecer 2´dGTP como substrato. A análise da cinética enzimática de XAC0610 na presença de 2´dGTP e GTP mostrou a formação preferencial do produto linear híbrido pppGp2´dG. O segundo capítulo aborda estudos sobre o metabolismo do cianeto em Bacillus com foco na cianeto dihidratase de Bacillus safensis. O cianeto é amplamente utilizado nas indústrias devido à sua alta afinidade com os metais. Esta mesma capacidade confere toxicidade potente a este composto. Assim, as indústrias têm que reduzir a concentração de cianeto das águas residuais antes de sua disposição final. Métodos físicos, químicos e biológicos têm sido desenvolvidos para atingir esse objetivo, mas o conhecimento sobre as vias metabólicas e a biologia das enzimas envolvidas na degradação do cianeto ainda é escasso. Aqui, é descrito o isolamento de uma cepa de Bacillus safensis de rejeitos de minas no Peru. A classificação desta cepa foi feita através de uma análise comparativa de 132 core genomes de cepas do grupo de Bacillus pumilus. Em seguida, determinamos que uma cianeto dihidratase (CynD, EC 3.5.5.1) codificada no genoma da cepa isolada era provavelmente a enzima responsável pela degradação do cianeto. A confirmação da atividade degradante de cianeto de CynD desta cepa foi feita por clonagem, expressão e purificação da enzima e realização de caracterização enzimática. O CynD desta cepa é ativo até pH 9 e os padrões de oligomerização analisados por SEC-MALS mostraram que a enzima forma longas estruturas helicoidais em pH 8 e estruturas menores enquanto o pH aumenta. Finalmente, foi demonstrado que a expressão de CynD é fortemente induzida na presença de cianeto. Os últimos dois anos do doutorado foram realizados no contexto da pandemia COVID- 19. Vários laboratórios se dedicaram a gerar conhecimento para ajudar no combate à pandemia. Nesta situação e graças à grande quantidade de dados genômicos disponíveis publicamente, estudos sobre a dinâmica das mutações do SARS-CoV-2 foram realizados. No primeiro ano da pandemia, a classificação genômica de 171.461 genomas mostrou a presença de cinco haplótipos principais com base em nove mutações. A distribuição mundial e a mudança de frequência desses haplótipos foram analisadas cuidadosamente. Todos os haplótipos foram identificados nas seis regiões analisadas (América do Sul, América do Norte, Europa, Ásia, África e Oceania); no entanto, a frequência de cada um deles foi diferente em cada uma dessas regiões. Em 30 de setembro de 2020, o haplótipo 3 (ou unidade taxonômica operacional 3, OTU_3) era o mais prevalente em quatro regiões (América do Sul, Ásia, África e Oceania). OTU_5 foi o mais prevalente na América do Norte e OTU_2 na Europa. A dinâmica temporal dos haplótipos mostrou que OTU_1 parece perto da extinção após 8 meses de pandemia (novembro de 2020). Outros OTUs ainda estão presentes em diferentes frequências em todo o mundo, mesmo atualmente gerando novas variantes. Com base em sua dinâmica temporal, um esquema de classificação de 115 mutações SARS-CoV-2 identificadas a partir de 1.058.020 genomas SARS-COV-2 também foi feito. Três tipos de dinâmica temporal de mutações foram identificados: i) Mutações de alta frequência, ii) mutações de média frequência e iii) mutações de baixa frequência. Finalmente, foi analisada a correlação do número de reprodução efetiva (Rt) do SARS-CoV-2 que contém a mutação de alta frequência N501Y com o nível de medidas de controle, mostrando que seu Rt está negativamente correlacionado com o nível de medidas de controle em oito dos nove países analisados. Esta correlação negativa foi semelhante quando foi analisado o Rt de SARS-CoV-2 sem a mutação N501Y. Assim, as medidas de controle provavelmente diminuirão o Rt de SARS-CoV-2 tipo selvagem e N501Y

Work process and hazard analysis in small-scale gold mining in Northern Philippines

Introduction@#Small-scale mining (SSM) is mining by individuals, groups, families, or cooperatives with minimal or no mechanization, often in the market's informal (illegal) sector. According to the Mines and Geosciences Bureau (MGB) in the Philippines, the gross production value of small-scale mining as of the 1st to 3rd quarter of 2020 was 0.5 billion pesos (1.05 billion USD). @*Objectives@#This study investigated the work process in small-scale mining in the northern part of the Philippines. It documented the occupational hazards that small-scale gold miners are exposed to in each of the work processes. @*Methods@#The target population is a community in the northern Philippines where the majority of the males are engaged in small-scale gold mining. This qualitative study used work observation and hazards analysis tools to investigate small-scale miners' work processes and hazard exposures. @*Results@#The most widely employed mining technique in the target community is dog-hole mining consisting of several sub-processes: tunneling, ball milling, and gravity concentration, cyanide leaching, and smelting. The occupational hazards identified were noise exposure from the dynamite blast, temperature extremes, and exposure to dust from dynamite blasting. The small-scale miners were subjected to prolonged crouching and bending, handling tools, and carrying heavy sacks filled with mineral ores. The miners resorted to improvised protective equipment such as wearing sleeveless shirts and drinking water for temperature extremes, distancing themselves from the mining blasts during dynamite blasting, and intermittently used carts with manual handling to transport ores packed in sacks. In the ball milling and gravity concentration process, machine-related accidents such as cuts from the crusher were observed. In cyanide leaching, which uses massive amounts of cyanide, the most prevalent hazards were heat, dust, and chemicals such as cyanide fumes. The risks included smoke from burning ore and coal and exposure to borax and nitric acid fumes in the smelting process. @*Conclusion@#This study documented the work process in small-scale gold mining and the hazard exposures in this type of informal industry. It is suggested that the local and national governments implement intervention programs for occupational health and safety measures, and social security nets are provided for the small-scale miners in Itogon, Benguet.

Intoxicación por cianuro como método suicida: reporte de caso / Cyanide poisoning as a suicide method: case report

Resumen Las intoxicaciones derivan de la presencia en el organismo de un tóxico o veneno, la muerte por intoxicación es una muerte violenta y por tanto requiere de la realización de una autopsia medico legal, la misma puede darse en el contexto de una exposición accidental ya sea en el hogar o laboral o sucitada por un intento de autoeliminación. La intoxicación por cianuro puede ser intencional (suicidio u homicidio) o accidental, los hallazgos en la autopsia medico legal son inespecíficos por lo que son importantes los datos aportados en el informe sobre muerte en investigación, el informe del escenario de muerte en caso de que un médico forense se hiciera presente al mismo y el resultado de los análisis toxicológicos, los cuales actualmente no se realizan en la sección de toxicología del poder judicial. Se realizó una revisión bibliográfica en diferentes bases de datos, de los artículos publicados referentes al tema de los últimos cinco años, con el objetivo de revisar las caracteristicas del químico, el metabolismo y la intoxicación como tal, tanto por sus secuelas como por sus implicaciones letales. Se concluye que para mejorar la pericia médico legal ante casos de intoxicacion por cianuro es fundamental conocer el mecanismo de acción y los posibles hallazgos presentes tanto al examen externo como interno, así como implementar que dicho escrutinio se incluya dentro del listado de sustancias a analizar.

Abstract Poisoning derives from the presence of a toxic substance or poison in the body, death by poisoning is a violent death and requires a legal medical autopsy, it may occur in the context of an accidental exposure at home or work, or caused by an attempt of self-elimination. Cyanide poisoning can be intentional (suicide / homicide) or accidental, the findings in the autopsy are unspecific, so data provided in the report of death in investigation, the report of the death scene (in case a forensic doctor was present) and the result of the toxicological analyzes, which are not currently performed in the toxicology section of the judiciary, are important. A bibliographic review was carried out in different databases of articles published in the last five years on the subject, with the objective of reviewing the chemical characteristics, the metabolism and the intoxication, as well, including their sequels and lethal implications. It is concluded that to improve the medical legal expertise in cases of cyanide poisoning, it is essential to know the mechanism of action and the possible findings in the external and internal examination; and to implement such scrutiny in the list of substances to be analyzed.

Intoxicación por cianuro, perspectiva desde urgencias: reporte de dos casos y revisión de la literatura / Cyanide poisoning, perspective from the emergency room: report of two cases and literature review / Intoxicação por cianeto, perspectiva desde urgências: relato de dois casos e revisão da literatura

Aunque poco común, la intoxicación por cianuro acarrea una mortalidad alta, sobre todo, cuando no se administra el tratamiento oportuno. La exposición a este tóxico puede darse por inhalación o por ingestión oral. Lo que se ha descrito sobre el manejo de esta intoxicación está basado en reportes de casos y pequeñas series, dado que en la literatura no se encuentran estudios que den un sustento claro. Dentro de estos reportes, y según los aspectos fisiopatológicos, son especialmente relevantes: el nitrito de sodio, el nitrito de amilo, el tiosulfito de sodio y la hidroxicobalamina, como arsenal terapéutico de esta intoxicación.

Cyanide intoxication is a rare condition. Yet, it leads to a high mortality, especially when proper treatment is not available. The exposure to this substance may occur via inhalation or oral ingestion. The basis of this intoxication´s treatment is on little case series and reports; this is due to the fact that the literature to support it remains scant and unclear. Based on physiopathological aspects, within this evidence the following are particularly relevant: sodium nitrile, amile nitrile, sodium tiosulphite, and hidroxicobalamin, as part of this intoxication´s treatment.

Embora pouco comum, a intoxicação por cianeto provoca uma mortalidade alta, sobre tudo, quando não se administra o tratamento oportuno. A exposição a este tóxico pode dar-se por inalação ou por ingestão oral. O que se há descrito sobre o manejo desta intoxicação está baseado em reportes de casos e pequenas séries, dado que na literatura não se encontram estudos que deem um sustento claro. Dentro destes relatos, e segundo os aspectos fisiopatológicos, são especialmente relevantes: o nitrito de sódio, o nitrito de amilo, o tiossulfato de sódio e a hidroxicobalamina, como arsenal terapêutico desta intoxicação.

Impacto percibido en la salud de los mineros artesanales del municipio de Quinchía (Colombia) por el uso de mercurio y cianuro en el proceso de amalgamiento de oro / Perceived impact on the artisanal miner's health from quinchias municipality (Colombia) by the use of cyanide and mercury in the amalgamation process of gold

RESUMEN Objetivo Determinar el impacto percibido en la salud de los mineros artesanales del municipio de Quinchía (Colombia) por el uso de mercurio y cianuro en el proceso de amalgamiento de oro. Métodos Investigación cuantitativa, descriptiva, en una muestra de 28 mineros activos expuestos a cianuro y mercurio. Se caracterizó el proceso de amalgamiento de oro, se realizó una encuesta de percepción de síntomas y se efectuaron exámenes de laboratorio para detectar la concentración de mercurio y cianuro en la sangre. Resultados El 50% de los mineros llevan en el oficio entre 21 y 30 años, con una exposición de 2,6 horas/día al mercurio y cianuro. El 33% de los mineros presentan niveles de concentración de mercurio en la sangre entre 4 y 4,9 microgramos por litro (μg/L) y refieren síntomas en los sistemas nervioso y digestivo. Los resultados de cianuro en la sangre fueron negativos. Conclusiones Las concentraciones de mercurio en la sangre se asocian con el tiempo en el oficio, horas de exposición al mercurio y percepción de síntomas. Los resultados de cianuro en la sangre no son coincidentes con los síntomas referidos por los mineros. Pese a que, se reconoce la toxicidad de estas sustancias químicas, aún se emplean métodos tradicionales y prohibidos sin tener en cuenta los protocolos de seguridad y el uso de elementos de protección adecuados. Los mineros constantemente están expuestos a procedimientos que implican riesgos para la salud; refieren síntomas no diagnosticados y que podrían afectar el organismo a largo plazo. No se adoptan medidas de prevención y promoción.(AU)

ABSTRACT Objective Determine the Perceived Impact on the Artisanal Miner's Health from Quinchía's Municipality (Colombia) by the Use of Cyanide and Mercury in the Amalgamation Process of Gold. Methods Quantitative, descriptive research in a sample of 28 active miners exposed to cyanide and mercury. The process of gold amalgamation was characterized, a survey of symptom perception was carried out and laboratory tests were carried out to detect the concentration of mercury and cyanide in blood. Results 50% of the miners are in the trade between 21 and 30 years, with an exposure of 2.6 hours/day to mercury and cyanide. 33% of miners have levels of mercury concentration in blood between 4-4.9 micrograms per liter (μg/L), as well as refer to symptoms in the nervous and digestive systems. The results of cyanide in blood are negative. Conclusions Blood mercury concentrations are associated with time in the trade, hours of exposure to mercury and perception of symptoms. The results of cyanide in blood are not coincident with the symptoms reported by the miners. Although the toxicity of these chemical substances is recognized, traditional and prohibited methods are still used without taking into account safety protocols and the use of appropriate protection elements. Miners are constantly exposed to procedures that involve health risks, refer undiagnosed symptoms and that could affect the organism in the long term. Prevention and promotion measures are not adopted.(AU)

Simultaneous biodegradation of phenol and cyanide present in coke-oven effluent using immobilized Pseudomonas putida and Pseudomonas stutzeri

ABSTRACT Discharge of coke-oven wastewater to the environment may cause severe contamination to it and also threaten the flora and fauna, including human beings. Hence before dumping it is necessary to treat this dangerous effluent in order to minimize the damage to the environment. Conventional technologies have inherent drawbacks however, biological treatment is an advantageous alternative method. In the present study, bacteria were isolated from the soil collected from the sites contaminated by coke-oven effluent rich in phenol and cyanide. Nucleotides sequence alignment and phylogenetic analysis showed the identity of the selected phenol and cyanide degrading isolates NAUN-16 and NAUN-1B as Pseudomonas putida and Pseudomonas stutzeri, respectively. These two isolates tolerated phenol up to 1800 mg L-1 and cyanide up to 340 mg L-1 concentrations. The isolates were immobilized on activated charcoal, saw dust and fly ash. The effluent was passed through the column packed with immobilized cells with a flow rate of 5 mL min-1. The isolates showed degradation of phenol up to 80.5% and cyanide up to 80.6% and also had the ability to reduce biological oxygen demand, chemical oxygen demand and lower the pH of effluent from alkaline to near neutral. The study suggests the utilization of such potential bacterial strains in treating industrial effluent containing phenol and cyanide, before being thrown in any ecosystem.

Hidroxocobalamina no tratamento de intoxicações por cianeto / Hydroxocobalamin in the treatment of cyanide poisoning

CONTEXTO: A intoxicação por cianeto pode ser considerada uma intoxicação rara porém de extrema gravidade. A causa mais comum de exposição aguda ao cianeto é a inalação de fumaça em incêndios. Nos casos de intoxicação, além das medidas de suporte clínico, como suplementação de oxigênio, a terapia com antídotos deve ser realizada. Dentre os antídotos disponíveis (hidroxocobalamina, nitrito de amila, nitrito de sódio, tiossulfato de sódio, 4-dimetilaminofenol e edetato de dicobalto), a hidroxocobalamina é apontada como o antídoto de primeira linha. Atualmente, tais medicamentos não estão disponíveis no Brasil. EVIDÊNCIAS CIENTÍFICAS: Dentre as melhores evidências recuperadas, após buscas por revisões sistemáticas, encontram-se 4 estudos observacionais realizados na França, sendo um de delineamento prospectivo e os demais retrospectivos (um total de 345 pacientes estudados). A maioria dos pacientes foi exposta ao cianeto por inalação de fumaça em incêndios domésticos, exceto por um estudo que avaliou principalmente tentativas de suicídio com ingestão de cianeto. Como intervenção, os estudos preconizaram uma dose inicial de 5g de hidroxocobalamina em infusão de 15-30 min, a qual foi administrada em aproximadamente 20 min após o contato com o serviço de emergência ainda em âmbito pré-hospitalar, com a possibilidade de doses adicionais em pacientes não responsivos (até um total de 15g). Como resultados, a mortalidade variou de 28-42% considerando todos os indivíduos que receberam a hidroxocobalamina. Entre os indivíduos com intoxicação confirmada laboratorialmente, 33-36% vieram a falecer, sendo poupados até mesmo indivíduos com níveis plasmáticos potencialmente letais, nestes a morte ocorreu em 36-39% dos casos. A parada cardíaca se apresentou como uma complicação comum (38%). A presença de sequelas no momento da alta hospitalar foi de 10-14%, sendo confusão, perda de memória e síndrome cerebelar as mais comuns. A hidroxocobalamina apresentou um perfil de segurança favorável, apenas com incidência de efeitos adversos leves. Dentre eles, o mais comum foi a apresentação de coloração vermelho-rosa na pele e urina e, mais raramente, aumento da pressão arterial. Após a avaliação crítica com a proposta do sistema GRADE, as evidências de eficácia atualmente disponíveis foram classificadas com qualidade muito baixa e as evidências de segurança com qualidade moderada. DISCUSSÃO: A hidroxocobalamina se apresenta como um agente potencialmente efetivo no tratamento de intoxicações por cianeto. Suas evidências devem ser interpretadas com cautela devido às limitações de suas fontes. O delineamento descritivo não permite o testes das variadas hipóteses, sem, portanto, ser quantificada a influência do acaso sobre os resultados obtidos. Da mesma forma, a ausência de controles e ajustes estatísticos não afasta a influência de fatores de confusão, sendo esses, importantes fontes de viés nos estudos observacionais. Com os custos considerados nas análises econômicas, ela se apresenta também como uma opção potencialmente custo-efetiva e com baixo impacto orçamentário. Todavia, outros fatores além da qualidade das evidências, como as barreiras para a sua devida implementação, devem ser considerados na elaboração de uma recomendação sobre seu uso. DECISÃO FINAL: Após as considerações provenientes da Consulta Pública, os membros da CONITEC presentes na 40ª reunião do plenário do dia 08/09/2015 deliberaram, por unanimidade, recomendar a incorporação do Cloridrato de hidroxocobalamina na concentração de 5 g injetável no tratamento de intoxicações por cianeto. Foi assinado o Registro de Deliberação nº 149/2015. DECISÃO: Incorporar a hidroxocobalamina no tratamento de intoxicações por cianeto, no âmbito do Sistema Único de Saúde ­ SUS, dada pela Portaria nº 9 de 28 de janeiro de 2016 publicado no DOU nº 20 de 29 de janeiro de 2016.

Farinha da casca do fruto de Passiflora edulis f. flavicarpa Deg (maracujá-amarelo): do potencial terapêutico aos efeitos adversos / Peel flour of the Passiflora edulis f. flavicarpa Deg (yellow passion fruit): From therapeutic potentials to side effects.

RESUMO O maracujá-amarelo (Passiflora edulis f. flavicarpa Deg) é considerado um alimento funcional, visto que, além de funções nutricionais, desempenha ações em vias metabólicas específicas. A farinha da casca do maracujá-amarelo (Passiflora edulis f. flavicarpa Deg) é rica em pectina. Esta fibra reduz a absorção de glicídios e lipídios, influenciando no metabolismo destes nutrientes. Portadores de patologias associadas às alterações no metabolismo glicêmico e lipidêmico têm utilizado este recurso de forma indiscriminada. Porém, substâncias designadas glicosídeos cianogênicos, presentes na casca do fruto são tóxicas ao organismo e prejudiciais à saúde. Ademais, o uso exacerbado de agrotóxicos na produção do maracujá é preocupante e a ingestão destes compostos também pode acarretar complicações à saúde. Portanto, o objetivo do presente trabalho foi sintetizar o conhecimento disponível referente aos efeitos terapêuticos e colaterais da suplementação com a farinha da casca do maracujá-amarelo (Passiflora edulis f. flavicarpa Deg). Foi realizada uma revisão bibliográfica nas bases de dados indexadas SCIELO, LILACS e PUBMED, sem limitação quanto ao período de publicação. Os estudos inclusos no trabalho, de forma unânime, apresentam eficácia da suplementação com a farinha da casca do maracujá-amarelo. A trituração da casca do fruto, realizada no processo de obtenção da farinha, é incapaz de promover redução de glicosídeos cianogênicos e agrotóxicos, permitindo que os indivíduos submetidos à suplementação estejam susceptíveis aos efeitos tóxicos destes compostos. Dessa forma, o desenvolvimento de estudos que avaliem a toxicidade da suplementação por períodos crônicos é de suma importância. Referente ao uso de agrotóxicos, a produção da farinha com frutos orgânicos pode ser uma alternativa para atenuar a toxicidade, sendo necessária a elucidação desta hipótese através de estudos.

ABSTRACT The yellow passion fruit (Passiflora edulis f. flavicarpa Deg) is considered a functional food, because, in addition to nutritional functions, it performs actions in specific metabolic pathways. The yellow passion fruit peel flour (Passiflora edulis f. flavicarpa Deg) is rich in pectin. This fiber reduces the absorption of carbohydrates and lipids, influencing the metabolism of these nutrients. Individuals affected by pathologies associated with alterations in the metabolism of lipids and carbohydrates have used this resource indiscriminately, however, substances called cyanogenic glycosides, present in the fruit peel, are toxic to the body and harmful to health. In addition, the overuse of pesticides in passion fruit production is becoming worrisome and the ingestion of these compounds can also cause negative health implications. Therefore, the aim of this study was to synthesize the available knowledge regarding the therapeutic effects and side effects of supplementation with the yellow passion fruit peel flour (Passiflora edulis f. flavicarpa Deg). A literature review was conducted from the indexed databases SCIELO, LILACS and PUBMED, without limitation to the publication period. The studies included in the work, unanimously, have effectiveness of supplementation with yellow passion fruit peel flour (Passiflora edulis f. flavicarpa Deg). The crushing of the fruit peel, performed in the process of obtaining flour, is unable to promote a reduction of cyanogenic glycosides and pesticides, causing individuals undergoing supplementation to be susceptible to the toxic effects of these compounds. Thus, this study’s development to evaluate the toxicity of supplementation for chronic periods is very important. With reference to the use of pesticides, flour production with organic fruits can be an alternative to attenuate toxicity and it is necessary to elucidate this hypothesis through this study.

Cyanide toxicity during cardiopulmonary bypass with small dose of nitroprusside: a case report / 대한마취과학회지

Sodium nitroprusside (SNP) is an anti-hypertensive drug, commonly used to decrease the systemic vascular resistance and lower the blood pressure. When the amount of cyanide generated by the SNP exceeds the metabolic capacity for detoxification, cyanide toxicity occurs. Under general anesthesia and cardiopulmonary bypass (CPB), it may be difficult to detect the development of cyanide toxicity. In cardiac surgical patients, hemolysis, hypothermia and decreased organ perfusion, which emphasize the risk of cyanide toxicity, may develop as a consequence of CPB. In particular, hemolysis during CPB may cause an unexpected overproduction of cyanide due to free hemoglobin release. We experienced a patient who demonstrated SNP tachyphylaxis and cyanide toxicity during CPB, even though the total amount of SNP administered was much lower than the recommended dose. We therefore report this case with a review of the relevant literature.

Aerobic cyanide degradation by bacterial isolates from cassava factory wastewater

Ten bacterial strains that utilize cyanide (CN) as a nitrogen source were isolated from cassava factory wastewater after enrichment in a liquid media containing sodium cyanide (1 mM) and glucose (0.2% w/v). The strains could tolerate and grow in cyanide concentrations of up to 5 mM. Increased cyanide levels in the media caused an extension of lag phase in the bacterial growth indicating that they need some period of acclimatisation. The rate of cyanide removal by the strains depends on the initial cyanide and glucose concentrations. When initial cyanide and glucose concentrations were increased up to 5 mM, cyanide removal rate increased up to 63 and 61 per cent by Bacillus pumilus and Pseudomonas putida. Metabolic products such as ammonia and formate were detected in culture supernatants, suggesting a direct hydrolytic pathway without an intermediate formamide. The study clearly demonstrates the potential of aerobic treatment with cyanide degrading bacteria for cyanide removal in cassava factory wastewaters.

Management of Cyanide Intoxication with Extracorporeal Membrane Oxygenation and Continuous Renal Replacement Therapy / 대한중환자의학회지

Cyanide intoxication results in severe metabolic acidosis and catastrophic prognosis with conventional treatment. Indications of extracorporeal membrane oxygenation (ECMO) and continuous renal replacement therapy (CRRT) are expanding to poisoning cases. A 50-year-old male patient arrived in the emergency room due to mental change after ingestion of cyanide as a suicide attempt 30 minutes prior. He was comatose, and brain stem reflexes were absent. Initial laboratory analysis demonstrated severe metabolic acidosis with increased lactic acid of 25 mM/L. Shock and acidosis were not corrected despite a large amount of fluid resuscitation with highdose norepinephrine and continuous renal replacement therapy. We decided to apply ECMO and CRRT to allow time for stabilization of hemodynamic status. After administration of antidote infusion, although the patient had the potential to progress to brain death status, vital signs were improved with correction of acidosis. We considered the evaluation for organ donation. We report a male patient who showed typical cyanide intoxication as lethal metabolic acidosis and cardiac impairment, and the patient recovered after antidote administration during vital organ support through ECMO and CRRT.

Management of Cyanide Intoxication with Extracorporeal Membrane Oxygenation and Continuous Renal Replacement Therapy / 대한구급학회지

Cyanide intoxication results in severe metabolic acidosis and catastrophic prognosis with conventional treatment. Indications of extracorporeal membrane oxygenation (ECMO) and continuous renal replacement therapy (CRRT) are expanding to poisoning cases. A 50-year-old male patient arrived in the emergency room due to mental change after ingestion of cyanide as a suicide attempt 30 minutes prior. He was comatose, and brain stem reflexes were absent. Initial laboratory analysis demonstrated severe metabolic acidosis with increased lactic acid of 25 mM/L. Shock and acidosis were not corrected despite a large amount of fluid resuscitation with highdose norepinephrine and continuous renal replacement therapy. We decided to apply ECMO and CRRT to allow time for stabilization of hemodynamic status. After administration of antidote infusion, although the patient had the potential to progress to brain death status, vital signs were improved with correction of acidosis. We considered the evaluation for organ donation. We report a male patient who showed typical cyanide intoxication as lethal metabolic acidosis and cardiac impairment, and the patient recovered after antidote administration during vital organ support through ECMO and CRRT.

Sulfane sulfur deficiency in malignant cells, increasing the inhibiting action of acetone cyanohydrin in tumor growth

PURPOSE: To demonstrate the irreversible poisoning action of the acetone cyanohydrin (AC) in malignant cells. METHODS: Thirty male Swiss mice were inoculated with 1x10³ Ehrlich tumor (ET) cells. The mice were divided into three groups (n=10): CG (saline); ACG1 (1.864 mg/Kg of AC) and ACG2 (2.796 mg/Kg of AC), treated every 48 hours from day 3 until day 13. On day 15 the mice were euthanized and the number of viable cells in ascites was determined. In the meantime, ET cells were incubated with AC (0.5, 1.0, 2.0 μg/mL). Cell viability and percentage of growth inhibition (PGI) were checked after one, two, three, four, 18 and 24 hours. RESULTS: There was reduction in volume and number of viable cells in ACG1 and ACG2 compared to CG. In ACG1 one of the animals did not present ascites. In ACG2 two mice did not present ascites and in CG none of the mice present ascites. The action of AC was dose and time dependent and there was no significant difference among the three doses. CONCLUSION: The acetone cyanohydrin promoted reduction of the tumor and also prevented tumor development in 20% of the treated animals.

Amygdalin inhibits angiogenesis in the cultured endothelial cells of diabetic rats.

Background: Angiogenesis contributes to different physiological and pathological conditions. The aim of this study was to investigate for the first time the antiangiogenic effects of amygdalin on the cultured endothelial cells of diabetic rats. Materials and Methods: A total of 20 streptozotocin-induced diabetic rats were divided into two equal groups of control and amygdalin-treated animals. Eight weeks after the induction of diabetes, amygdalin was injected intraperitoneally (3 mg/kg) to the rats of the treatment group. One day later, rats were sacrificed; the aortic arteries were excised and cut as 2 mm rings. Each aortic ring was incubated in a cell-culture well for 7 days. The process of angiogenesis was monitored by counting the number of microvessels and primary microtubules in each well. Results: Optic microscopy showed proliferation and migration of new endothelial cells to the fibrin gels. The endothelial cells produced primary microtubules which gradually made several branches and finally made a vascular matrix. The number of the primary microtubules and microvessels were significantly lower in the amygdalin-treated vs. control group (P < 0.01). Conclusion: Therefore, amygdalin exerts inhibitory effects on angiogenesis in aortic rings of diabetic rats and may pave a new way for treatment of unfavorable angiogenic conditions.

Toxic effects of zinc cyanide on some protein metabolites in fresh water fish, Cirrhinus mrigala [hamilton]

Effect of zinc cyanide on protein and certain metabolites content and some enzymes activities was evaluated in liver, muscle and gill tissues of the freshwater fish, Cirrhinus mrigala, during exposure and following cessation of exposure. Fish exhibited significant alterations in the biochemical parameters in all tissues studied. Levels of total protein decreased in all tissues, where as free amino acids [FAA] and protease activities significantly increased [P<0.05]. Similarly decrease was observed in the ammonia level with increased urea and glutamine levels at all exposure periods. The enzymes involved in the protein metabolism altered under the zinc cyanide intoxication. Increase in the protease and aminotransferases revealed amplified transamination processes. Significant increase of phosphatases indicated increased rate of phosphorylation and transport of molecules across the cell membrane. Withdrawal study also exhibited significant recovery in all above biochemical parameters, in all tissue after the 7th day post recovery treatment. Present study exhibited negative effects of zinc cyanide on protein metabolism. Fish with low protein content were not fit for human consumption. Patterns of effects on intermediary metabolism suggest avenues to determine the mechanisms by which such effects occur

Measurement of acetonitrile in blood and urine by head-space gas chromatography / 法医学杂志

OBJECTIVE@#To establish the method for measurement of acetonitrile in blood and urine by head-space gas chromatography.@*METHODS@#DB-ALC1 (30 m x 320 microm x 1.8 microm) and DB-ALC2 (30 m x 320 microm x 1.2 microm) capillary column were used to measure the acetonitrile in blood and urine with the isopropanol as internal standard reference.@*RESULTS@#The limits of detection of acetonitrile in both blood and urine were 0.5 microg/mL, with a linear range of 5-1000 microg/mL (r = 0.999).The accuracy of this method was 93.2%-98.0%. The RSD for the intra-day and inter-day were less than 3.7%.@*CONCLUSION@#The method is capable for measurement analysis of acetonitrile in blood and urine.

Biotratamiento de residuos cianurados y su relación con la salud pública / Cyanide residues biotreatment and their relation with public health

Objetivo: Proponer un biotratamiento para los residuos cianurados generados por procedimientos medico-forenses a identificar los factores de riesgo procedentes de su manipulación. Metodología: Los residuos de cianuro procedentes del Instituto de Medicina legal, sede Medellín, se caracterizaron según su estado físico y determinaron su manejo, y los factores de riesgo. Para la degradación de los residuos se realizaron preensayos y 9 bioensayos con Pseudomonas aeruginosa y thiobacillus ferrooxidans para diferentes concentraciones de cianuro. Resultados: Se observa deficiencia en todos los factores de riesgo analizados, siendo el componente químico el que predominó en fuentes de contaminación. En los ensayos de degradación, Pseudomonas aeruginosa demostró un porcentaje de remoción del 87% para una concentración 50mg/L y thiobacillus ferrooxidans, un 79% de remoción para una concentración de 500mg/L. Se realizó una guía técnica para el manejo de los residuos de cianuro. Conclusiones: En la evaluación de las rutas de cianuro, desde la incautación hasta su disposición final, se encontró que el Instituto de Medicina Legal no contaba con pautas documentadas sobre la gestión de éstos. Los tratamientos biológicos con bacterias como Pseudomonas aeruginosa y thiobacillus ferrooxidans fueron eficientes en la degradación de cianuro.

Objective: to propose a bio-treatment for cyanide residues generated by medico-legal procedures and to identify risk factors from handling. Methodology: cyanide residues, from the Institute of Legal Medicine, Medellin, were characterized by their physical state and identified its management, and risk factors. For the residues degradation 22 pre-essays and 9 bioassays were done with Pseudomonas aeruginosa and thiobacillus ferrooxidans at different cyanide concentrations. Results: there were failures in all risk factors analyzed, predominating the chemical component which sources of pollution. In studies of degradation, Pseudomonas aeruginosa showed a 87% removal percentage at a concentration of 50mg/L and thiobacillus ferrooxidans, 79% removal at a concentration of 500mg/L. We conducted a technical guide for the management of cyanide residues. Conclusions: The evaluation of the routes of cyanide from seizure to final disposal, were found that the Institute of Forensic Medicine had no documented guidelines on the management of them. Biological treatment with bacteria such as Pseudomonas aeruginosa and thiobacillus ferrooxidans were efficient in the degradation of cyanide.