Production of extracellular L-asparaginase : from bioprospecting to the engineering of an antileukemic biopharmaceutical

The L-asparaginase (L-asparagine amino hydrolase, E.C.3.5.1.1) catalyzes the hydrolysis of L-aspargine into L-aspartic acid and ammonia. The effective depletion of L-asparine results in cytotoxicity for leukemic cell. Therefore the enzyme has been a clinically acceptable anti-tumour agent for the effective treatment of acute lymphoblastic leukemia (ALL) and lymphosarcoma. L-asparaginase production using microbial system had attracted considerable attention, owing to the cost effective and eco friendly nature. A wide range of microorganisms such as filamentous fungi, yeasts and bacteria have proved to be the good sources of the enzyme L-asparaginase. Thus, in this review mainly focuses on the biochemical aspects of L-asparaginase production, aiming to comprehend the physiochemical characteristics, such as stability, bioavailability, toxicity, allergenic aspects, application, and enzyme properties and kinetics of recombinant enzyme production by fermentation. Processes central to these biochemical aspects, including fermentation of L-asparaginase producing organisms and downstream processing of the enzyme are also discussed

Differentially regulated induced resistance marker enzymes in soybean genotypes resistant and susceptible to Asian soybean rust

O objetivo deste trabalho foi avaliar, por meio de enzimas marcadoras, a indução de resistência à ferrugem‑asiática‑da‑soja em genótipos de soja contrastantes quanto à suscetibilidade a Phakopsora pachyrhizi. A proteína total e as atividades de cinco enzimas marcadoras da indução de resistência (lipoxigenases, peroxidases, fenilalanina amônia‑liase, quitinases e β‑1,3‑glucanases) foram avaliadas em extratos de folhas de plantas de soja dos genótipos Embrapa 48 (suscetível) e PI 561356 (resistente), submetidas à inoculação ou não com o patógeno. Foram observadas respostas de defesa discrepantes entre os dois genótipos e entre os tempos de coleta (12, 72 e 168 horas após inoculação). A resposta de indução dessas enzimas assemelha-se à defesa bifásica, para Embrapa 48, e é consistente com o observado para outros patossistemas. No entanto, o genótipo PI 561356 respondeu com diminuição da concentração de proteína total e das atividades enzimáticas, o que indica reduçãodo metabolismo geral das plantas infectadas. Há um importante mecanismo de resistência do genótipo PI 561356, ainda não relatado, embasado em vias que envolvem essas enzimas marcadoras e em mecanismos que utilizam menor concentração de proteínas, como os de via metabólica de resposta em cascata.The objective of this work was to evaluate induced resistance to Asian soybean rust by means of enzymeactivities in soybean genotypes contrasting as to their susceptibility to Phakopsora pachyrhizi. Total protein and the activities of five induced resistance marker enzymes (lipoxygenases, peroxidases, phenylalanine ammonia‑lyase, chitinases and β‑1,3‑glucanases) were evaluated in leaf extracts of soybean plants of the genotypes Embrapa 48 (susceptible) and PI 561356 (resistant), inoculated or not with the pathogen. Discrepant defense responses were obtained between the two genotypes and among the leaf harvest times (12, 72, and 168 hours after inoculation). The induction response of these enzymes resembles the biphasic defense in Embrapa 48 and is consistent with that observed in other pathological systems. However, the genotype PI 561356 responded with a decrease in total protein concentration and in enzymatic activities, indicating a general reduction in the metabolism of the infected plants. There is an important mechanism of resistance for the genotype PI 561356, not yet reported, which is grounded on the metabolic ways involving these induced resistance marker enzymes and on the mechanisms that use lower concentrations of total protein, such as the ones with metabolic pathways in response cascade

Enzimas marcadoras de indução de resistência diferencialmente reguladas em soja resistente e suscetível à ferrugem-asiática-da-soja

O objetivo deste trabalho foi avaliar, por meio de enzimas marcadoras, a indução de resistência à ferrugem-asiática-da-soja em genótipos de soja contrastantes quanto à suscetibilidade a Phakopsora pachyrhizi. Aproteína total e as atividades de cinco enzimas marcadoras da indução de resistência (lipoxigenases, peroxidases, fenilalanina amônia-liase, quitinases e β-1, 3-glucanases) foram avaliadas em extratos de folhas de plantas de soja dos genótipos Embrapa 48 (suscetível) e PI 561356 (resistente), submetidas à inoculação ou não com o patógeno. Foram observadas respostas de defesa discrepantes entre os dois genótipos e entre os tempos de coleta (12, 72 e 168 horas após inoculação). A resposta de indução dessas enzimas assemelha-se à defesa bifásica, para Embrapa 48, e é consistente com o observado para outros patossistemas. No entanto, o genótipo PI 561356 respondeu com diminuição da concentração de proteína total e das atividades enzimáticas, o que indica redução do metabolismo geral das plantas infectadas. Há um importante mecanismo de resistência do genótipo PI 561356, ainda não relatado, embasado em vias que envolvem essas enzimas marcadoras e em mecanismos que utilizam menor concentração de proteínas, como os de via metabólica de resposta em cascata.The objective of this work was to evaluate induced resistance to Asian soybean rust by means of enzyme activities in soybean genotypes contrasting as to their susceptibility to Phakopsora pachyrhizi. Total protein and the activities of five induced resistance marker enzymes (lipoxygenases, peroxidases, phenylalanine ammonia-lyase, chitinases and β-1, 3-glucanases) were evaluated in leaf extracts of soybean plants of the genotypes Embrapa 48 (susceptible) and PI 561356 (resistant), inoculated or not with the pathogen. Discrepant defense responses were obtained between the two genotypes and among the leaf harvest times (12, 72, and 168 hours after inoculation). The induction response of these enzymes resembles the biphasic defense in Embrapa 48 and is consistent with that observed in other pathological systems. However, the genotype PI 561356 responded with a decrease in total protein concentration and in enzymatic activities, indicating a general reduction in the metabolism of the infected plants. There is an important mechanism of resistance for the genotype PI 561356, not yet reported, which is grounded on the metabolic ways involving these induced resistance marker enzymes and on the mechanisms that use lower concentrations of total protein, such as the ones with metabolic pathways in response cascade

NEOTROPICAL XENARTHRANS: a data set of occurrence of xenarthran species in the Neotropics

Xenarthrans – anteaters, sloths, and armadillos – have essential functions for ecosystem maintenance, such as insect control and nutrient cycling, playing key roles as ecosystem engineers. Because of habitat loss and fragmentation, hunting pressure, and conflicts with 24 domestic dogs, these species have been threatened locally, regionally, or even across their full distribution ranges. The Neotropics harbor 21 species of armadillos, ten anteaters, and six sloths. Our dataset includes the families Chlamyphoridae (13), Dasypodidae (7), Myrmecophagidae (3), Bradypodidae (4), and Megalonychidae (2). We have no occurrence data on Dasypus pilosus (Dasypodidae). Regarding Cyclopedidae, until recently, only one species was recognized, but new genetic studies have revealed that the group is represented by seven species. In this data-paper, we compiled a total of 42,528 records of 31 species, represented by occurrence and quantitative data, totaling 24,847 unique georeferenced records. The geographic range is from the south of the USA, Mexico, and Caribbean countries at the northern portion of the Neotropics, to its austral distribution in Argentina, Paraguay, Chile, and Uruguay. Regarding anteaters, Myrmecophaga tridactyla has the most records (n=5,941), and Cyclopes sp. has the fewest (n=240). The armadillo species with the most data is Dasypus novemcinctus (n=11,588), and the least recorded for Calyptophractus retusus (n=33). With regards to sloth species, Bradypus variegatus has the most records (n=962), and Bradypus pygmaeus has the fewest (n=12). Our main objective with Neotropical Xenarthrans is to make occurrence and quantitative data available to facilitate more ecological research, particularly if we integrate the xenarthran data with other datasets of Neotropical Series which will become available very soon (i.e. Neotropical Carnivores, Neotropical Invasive Mammals, and Neotropical Hunters and Dogs). Therefore, studies on trophic cascades, hunting pressure, habitat loss, fragmentation effects, species invasion, and climate change effects will be possible with the Neotropical Xenarthrans dataset

Diretriz da Sociedade Brasileira de Cardiologia sobre Diagnóstico e Tratamento de Pacientes com Cardiomiopatia da Doença de Chagas

This guideline aimed to update the concepts and formulate the standards of conduct and scientific evidence that support them, regarding the diagnosis and treatment of the Cardiomyopathy of Chagas disease, with special emphasis on the rationality base that supported it.  Chagas disease in the 21st century maintains an epidemiological pattern of endemicity in 21 Latin American countries. Researchers and managers from endemic and non-endemic countries point to the need to adopt comprehensive public health policies to effectively control the interhuman transmission of T. cruzi infection, and to obtain an optimized level of care for already infected individuals, focusing on diagnostic and therapeutic opportunistic opportunities.   Pathogenic and pathophysiological mechanisms of the Cardiomyopathy of Chagas disease were revisited after in-depth updating and the notion that necrosis and fibrosis are stimulated by tissue parasitic persistence and adverse immune reaction, as fundamental mechanisms, assisted by autonomic and microvascular disorders, was well established. Some of them have recently formed potential targets of therapies.  The natural history of the acute and chronic phases was reviewed, with enhancement for oral transmission, indeterminate form and chronic syndromes. Recent meta-analyses of observational studies have estimated the risk of evolution from acute and indeterminate forms and mortality after chronic cardiomyopathy. Therapeutic approaches applicable to individuals with Indeterminate form of Chagas disease were specifically addressed. All methods to detect structural and/or functional alterations with various cardiac imaging techniques were also reviewed, with recommendations for use in various clinical scenarios. Mortality risk stratification based on the Rassi score, with recent studies of its application, was complemented by methods that detect myocardial fibrosis.  The current methodology for etiological diagnosis and the consequent implications of trypanonomic treatment deserved a comprehensive and in-depth approach. Also the treatment of patients at risk or with heart failure, arrhythmias and thromboembolic events, based on pharmacological and complementary resources, received special attention. Additional chapters supported the conducts applicable to several special contexts, including t. cruzi/HIV co-infection, risk during surgeries, in pregnant women, in the reactivation of infection after heart transplantation, and others.     Finally, two chapters of great social significance, addressing the structuring of specialized services to care for individuals with the Cardiomyopathy of Chagas disease, and reviewing the concepts of severe heart disease and its medical-labor implications completed this guideline.Esta diretriz teve como objetivo principal atualizar os conceitos e formular as normas de conduta e evidências científicas que as suportam, quanto ao diagnóstico e tratamento da CDC, com especial ênfase na base de racionalidade que a embasou. A DC no século XXI mantém padrão epidemiológico de endemicidade em 21 países da América Latina. Investigadores e gestores de países endêmicos e não endêmicos indigitam a necessidade de se adotarem políticas abrangentes, de saúde pública, para controle eficaz da transmissão inter-humanos da infecção pelo T. cruzi, e obter-se nível otimizado de atendimento aos indivíduos já infectados, com foco em oportunização diagnóstica e terapêutica. Mecanismos patogênicos e fisiopatológicos da CDC foram revisitados após atualização aprofundada e ficou bem consolidada a noção de que necrose e fibrose sejam estimuladas pela persistência parasitária tissular e reação imune adversa, como mecanismos fundamentais, coadjuvados por distúrbios autonômicos e microvasculares. Alguns deles recentemente constituíram alvos potenciais de terapêuticas. A história natural das fases aguda e crônica foi revista, com realce para a transmissão oral, a forma indeterminada e as síndromes crônicas. Metanálises recentes de estudos observacionais estimaram o risco de evolução a partir das formas aguda e indeterminada e de mortalidade após instalação da cardiomiopatia crônica. Condutas terapêuticas aplicáveis aos indivíduos com a FIDC foram abordadas especificamente. Todos os métodos para detectar alterações estruturais e/ou funcionais com variadas técnicas de imageamento cardíaco também foram revisados, com recomendações de uso nos vários cenários clínicos. Estratificação de risco de mortalidade fundamentada no escore de Rassi, com estudos recentes de sua aplicação, foi complementada por métodos que detectam fibrose miocárdica. A metodologia atual para diagnóstico etiológico e as consequentes implicações do tratamento tripanossomicida mereceram enfoque abrangente e aprofundado. Também o tratamento de pacientes em risco ou com insuficiência cardíaca, arritmias e eventos tromboembólicos, baseado em recursos farmacológicos e complementares, recebeu especial atenção. Capítulos suplementares subsidiaram as condutas aplicáveis a diversos contextos especiais, entre eles o da co-infecção por T. cruzi/HIV, risco durante cirurgias, em grávidas, na reativação da infecção após transplante cardíacos, e outros.    Por fim, dois capítulos de grande significado social, abordando a estruturação de serviços especializados para atendimento aos indivíduos com a CDC, e revisando os conceitos de cardiopatia grave e suas implicações médico-trabalhistas completaram esta diretriz.&nbsp

Proteomic analysis of Aedes aegypti midgut during post-embryonic development and of the female mosquitoes fed different diets

In this work we analyzed protein expression in the Aedes aegypti midgut during the larval (fourth instar, L4), pupal, and adult stages [including newly emerged (NE), sugar-fed (SF) and blood-fed (BF) females]. Two-dimensional electrophoresis showed 13 spots in the midgut of larvae, 95 in the midgut of pupae, 90 in the midgut of NE, and 76 in the midgut of SF or BF females. In the larval midguts, high serpin expression was noted, while in the pupae, protein abundance was lower than in the NE, SF, and BF females. The spots related to proteins linked to energy production, protein metabolism, signaling, and transport were highly expressed in the NE stage, while spots related proteins involved in translation were abundant in SF and BF females. The differential abundance of proteins in the midgut of A. aegypti at different developmental stages supports the necessity for midgut development during immature stage followed by the necessity of proteins related to digestion in adults

Plantas de pimentão submetidas à injúria mecânica modificam a expressão de proteínas em plantas vizinhas não injuriadas

DOI: não consta, por isso foi disponibilizado o link de acesso.A proteômica avalia eventos biológicos por análise das proteínas expressas em células ou tecidos em situações fisiológicas diferentes. Na proteômica de plantas, visando manter padrões de cultivo, plantas tratadas e não tratadas são muitas vezes cultivadas no mesmo ambiente, sem considerar alterações por plantas tratadas sobre a expressão de proteínas de plantas não tratadas. O objetivo desse trabalho foi, por proteômica, avaliar a expressão diferencial de proteínas em folhas de plantas de pimentão (Capsicum annuum L. ́Magali R‘) não-feridas (não-tratadas) e feridas (tratadas) por injúria mecânica, quando cultivadas ou não próximas fisicamente. Aos 40 dias após o plantio, parte das plantas foi submetida à injúria por ferimentos nas folhas totalmente expandidas (feridas, F), e parte foi reservada como plantas não-feridas, ou cultivadas perto das plantas F (NFP, a 0,15 m) ou cultivadas longe das plantas F (NFL, a 10 m). As folhas das plantas NFP e NFL foram coletadas às 12, 48 e 168 h após ferimentos das plantas F, e os extratos NFL e NFP foram analisados por eletroforese bidimensional (2-DE). Os perfis proteicos foram comparados por Image Master, e as proteínas identificadas usando espectrometria de massas. Houve menor concentração de proteína em extratos de NFP 12 h; em 48 h e 168 h, a proteína foi semelhante entre NFL e NFP. Por 2-DE, não houve diferenças significativas entre NFP e NFL 12 h, porém para NFP 48 h foi observada maior expressão de proteínas envolvidas na defesa de plantas. Para NFP 168 h, houve aumento da expressão de proteínas envolvidas no metabolismo normal. Esses resultados indicam ter havido respostas de defesa das plantas NFP, decorrentes da proximidade do cultivo dessas plantas com as plantas F, possivelmente por ação de voláteis. Coloca-se assim a importância da escolha das condições de cultivo dos tratamentos-controle, não apenas para análises proteômicas.Proteomics evaluates biological events by analysis of the expression of proteins in cells or tissues under different physiological situations. For plant proteomics, in order to maintain similar cultivation conditions, treated and non-treated plants are generally cultivated under the same environment without considering changes by the treated plants in protein expression of non-treated neighboring plants. The aim of this work was, by proteomics, assess the differential expression of proteins in non-wounded (non-treated) and wounded (treated) leaves of bell pepper (Capsicum annuum L. ‘Magali R’) by mechanical injury, when plants were grown physically nearby or not. At 40 days of cultivation, some plants were wounded by mechanical injury in all expanded leaves (wounds, F), some plants were maintained as non-injured plants, or being grown near to F plants (NFP, 0.15 m ) or being grown away from F (NFL, 10 m). NFP and NFL plant leaves were harvested at 12, 48, and 168 h after injury of F plants, and the NFL and NFP extracts were analyzed by two-dimensional electrophoresis (2-DE). The protein profiles were compared using Image Master and the proteins were identified using mass spectrometry. NFP 12 h extract presented a lower protein concentration; for 48 h and 168 h, the protein was similar for NFL and NFP. For 2-DE, no significant difference was observed between NFP 12 h and NFL 12 h. However, NFP 48 h showed a higher expression of proteins involved in plant defense. For NFP 168 h, an increased expression of proteins involved in normal metabolism was detected. These results indicated that defense responses were developed by NFP plants considering the proximity of non-wounded plants from wounded plants during cultivation, with volatile compounds involvement. In this sense, it should be emphasized the importance of the choice of the growth conditions for the control-treatments, not only for proteomics

Enzimas marcadoras de indução de resistência diferencialmente reguladas em soja resistente e suscetível à ferrugem-asiática-da-soja

O objetivo deste trabalho foi avaliar, por meio de enzimas marcadoras, a indução de resistência à ferrugem-asiática-da-soja em genótipos de soja contrastantes quanto à suscetibilidade a Phakopsora pachyrhizi. Aproteína total e as atividades de cinco enzimas marcadoras da indução de resistência (lipoxigenases, peroxidases, fenilalanina amônia-liase, quitinases e β-1, 3-glucanases) foram avaliadas em extratos de folhas de plantas de soja dos genótipos Embrapa 48 (suscetível) e PI 561356 (resistente), submetidas à inoculação ou não com o patógeno. Foram observadas respostas de defesa discrepantes entre os dois genótipos e entre os tempos de coleta (12, 72 e 168 horas após inoculação). A resposta de indução dessas enzimas assemelha-se à defesa bifásica, para Embrapa 48, e é consistente com o observado para outros patossistemas. No entanto, o genótipo PI 561356 respondeu com diminuição da concentração de proteína total e das atividades enzimáticas, o que indica redução do metabolismo geral das plantas infectadas. Há um importante mecanismo de resistência do genótipo PI 561356, ainda não relatado, embasado em vias que envolvem essas enzimas marcadoras e em mecanismos que utilizam menor concentração de proteínas, como os de via metabólica de resposta em cascata