A strategy for the rapid identification of fungal metabolites and the discovery of the antiviral activity of pyrenocine a and harzianopyridon

The isolation and identification of bioactive metabolites from complex extracts obtained from microbial growth media is a time consuming, costly, and labor-intensive task. A strategy to rapidly identify secondary metabolites isolated from extracts obtained from the culture media of marine-derived and endophytic fungal strains is described. Identification was achieved by HPLC-UV-MS and 1H NMR analyses in combination with data obtained from the Dictionary of Natural Products. Among the compounds identified, (-)-naphthoquinoneimine, citreorosein, emodin, pyrenocine A and harzianopyridone displayed moderate to potent antiviral activity. (-)-Naphthoquinoneimine was isolated as the enantiomer of its previously reported dextrorotatory congener, while 6,7-dihydroxy-2,2-dimethyl-4-chromanone is herein reported for the first time as a natural product396720731CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESPBEX 4498-14-32010/50190-2; 2013/50228-8; 2011/08064-2; 2008/00331-9; 2013/23153-

Discovery and biosynthetic studies of non-ribosomal peptides involved in Pseudovibrio brasiliensis Ab134 motility

Neste trabalho foram investigados metabólitos envolvidos na motilidade da α-Proteobactéria marinha Pseudovibrio brasiliensis Ab134, utilizando-se dados de sequenciamento do genoma, experimentos de inativação gênica e técnicas de cultivo em laboratório. O uso de análises de bioinformática permitiu identificar um gene cluster de biossíntese mista de peptídeo não-ribossomal sintetases e policetídeo sintase conservado em linhagens de Pseudovibrio spp. e Pseudomonas spp.. A inativação de genes por recombinação homóloga em P. brasiliensis Ab134, levou a descoberta dos heptapeptídeos pseudovibriamidas A1-6 e dos depsipeptídeos pseudovibriamidas B1-6, codificados por esse gene cluster. Estes compostos apresentam uma conexão via uma ureia, ao invés da uma ligação peptídica conectando os primeiros dois resíduos de aminoácidos. Outras modificações incluem um resíduo de ácido desidroaminobutírico, e um heterociclo tiazol. Bioensaios de motilidade mostraram que o mutante pppA, o qual não produz nenhum dos compostos, tem sua habilidade de swarm reduzida e formação de biofilme aumentada. Já os mutantes pppD, produtores apenas das pseudovibriamidas A, mantiveram sua habilidade de swarm, porém a formação de biofilme foi reduzida. Estes experimentos indicaram que os peptídeos produzidos por P. brasiliensis Ab134 podem atuar nos fenótipos relacionados à colonização de seu hospedeiro, a esponja marinha Arenosclera brasiliensis.The present study investigated the metabolites involved on Pseudovibrio brasiliensis Ab134 motility, a marine alpha-Proteobacteria, by using genome sequencing data, gene inactivation experiments and laboratory cultivation techniques. Bioinformatics analyzes allowed the identification of a mixed biosynthetic gene cluster of non-ribosomal peptide syntethases and polyketide synthase conserved in Pseudovibrio and Pseudomonas strains. Gene knock-out in P. brasiliensis Ab134 led the discovery of the compounds encoded by this cluster gene, the heptapeptides pseudovibriamide A1-6 and the depsipeptides pseudovibriamides B1-6. These compounds have a ureido-linkage instead of a peptide bond connecting the first two amino acid residues. Other modifications include a dehydroaminobutyric acid residue, and a thiazole heterocycle. Motility bioassays have shown that the pppA mutant, which does not produce any of the compounds, has reduced swarm motility and increased biofilm formation. The pppD mutants, which produce only pseudovibriamides A, maintained their swarm phenotype, but biofilm formation was impaired. These experiments suggest that the peptides produced by P. brasiliensis Ab134 can influence on phenotypes related to the colonization of the eukaryotic host, the marine sponge Arenosclera brasiliensis

Study of the metabolism of fungi using isotopically 13C-labeled precursors

Este trabalho objetivou o estudo de rotas de formação de metabólitos secundários utilizando precursores isotopicamente marcados com 13C. Os experimentos de crescimento com adição de [1-13C]acetato, [1,2-13C2]acetato e [U-13C315N1]-L-cisteína para o fungo do ambiente marinho Penicillium sp. DRF2 mostrou que as ciclotiocurvularinas são provenientes da rota de formação de policetídeos e pela incorporação de L-cisteína, depois da transformação desta em 3-mercaptopiruvato. Os resultados sugerem que a formação das ciclotiocurvularinas provém de um processo de detoxificação da α,β-desidrocurvularina. O estudo do metabolismo secundário de Aspergillus sp. DLM3-8, também do ambiente marinho, mostrou que o seu perfil metabólico produzido em experimentos de crescimento sob diferentes condições é constante. Os experimentos de incorporação de precursores isotopicamente marcados com 13C na naftoquinonaimina, produzida por Aspergillus sp. DLM3-8 foram inconclusivos, indicando que outras abordagens experimentais devem ser realizadas para se investigar a biossíntese deste metabólito.This investigation aimed investigated the formation routes of secondary metabolites using 13C-labelled precursors. Feeding experiments with [1-13C]acetate, [1,2-13C2]acetate and [U-13C315N1]-L-cysteine within the growth medium of the marine-derived fungi Penicillium sp. DRF2 showed that cyclothiocurvularins are derived from polyketides and from the incorporation of a L-cysteine residue, after its transformation into 3-mercaptopyruvate. The results suggest that the formation of cyclothiocurvularins is derived from a detoxification process ofα,β-dehydrocurvularin. Investigation of the secondary metabolism of a marine-derived Aspergillus sp. DLM3-8 indicated a stable metabolic profile under a variety of growth conditions. Feeding experiments with 13C-labelled precursors for the biosynthesis investigation of naphthoquinoneimine were inconclusive, indicating that other methodologies should be envisaged in order to investigate the biosynthesis of this metabolite

Natural Products from Marine Invertebrates and Microorganisms in Brazil between 2004 and 2017: Still the Challenges, More Rewards

<div><p>The Brazilian marine biodiversity represents a unique, yet underexplored resource of biologically active compounds. This review provides an analysis of the development of marine natural products chemistry in Brazil within the period comprised between 2004 and 2017. Emphasis is directed towards marine invertebrate and marine microorganisms metabolites, including isolation, structure analysis, biosynthesis, bioactivities and total synthesis. An overview of the research on marine natural products by Brazilian researchers is also discussed, as well as perspectives for the development of the chemistry of marine natural products in Brazil.</p></div

Características demográficas e incidência de fatores de risco associados à Hipertensão Arterial na 17ª SAFE em Araraquara-SP

Hypertension is a multifactorial clinical condition characterized by chronic high levels of arterial blood pressure (BP). It has extensive prevalence, high risk of mortality and morbidity and need for constant control and monitoring for therapeutic success. Based on this, students of pharmacy at UNESP structured an event in order to advise the population of the city of Araraquara about the risk factors and measure the people’s BP. This study aimed to identify the demographic characteristics and the incidence of risk factors for hypertension in the assisted population. Our data indicate that there are large numbers of individuals with high BP mainly in people diagnosed with hypertension, suggesting possible treatment failure. Moreover, men seem to have the BP less controlled than women and smoking and family history have been associated to the diagnosis of hypertension. Also, the frequent use of alcohol was correlated with high BP both in persons already diagnosed with hypertension, as undiagnosed.A hipertensão arterial é uma condição clínica multifatorial caracterizada por níveis crônicos elevados de pressão sanguínea arterial (PA). Ela possui ampla prevalência, elevado risco de mortalidade e morbidade e necessidade de constante controle e acompanhamento para o sucesso terapêutico. Com base nisso, os alunos do curso de farmácia-bioquímica da UNESP realizaram um evento com o intuito de alertar a população da cidade de Araraquara sofre os fatores de risco e fazer a aferição da pressão arterial. Esse trabalho visou identificar as características demográficas e a incidência dos fatores de risco para Hipertensão Arterial na população atendida. Nossos dados indicam que há um grande número de indivíduos com PA elevada principalmente em pessoas diagnosticadas com hipertensão, sugerindo possível falha no tratamento. Além disso, homens apresentam a PA menos controlada que mulheres e o tabagismo e histórico familiar apresentaram-se associados ao diagnóstico de hipertensão arterial. Além disso, o consumo frequente de bebidas alcoólicas correlaciona-se com a PA elevada tanto em indivíduos já diagnosticados com hipertensão, como os não diagnosticados

Early Oxidative Transformations During the Biosynthesis of Terrein and Related Natural Products

The mycotoxin terrein is derived from the C(10)‐precursor 6‐hydroxymellein (6‐HM) via an oxidative ring contraction. Although the corresponding biosynthetic gene cluster (BGC) has been identified, details of the enzymatic oxidative transformations are lacking. Combining heterologous expression and in vitro studies we show that the flavin‐dependent monooxygenase (FMO) TerC catalyzes the initial oxidative decarboxylation of 6‐HM. The reactive intermediate is further hydroxylated by the second FMO TerD to yield a highly oxygenated aromatic species, but further reconstitution of the pathway was hampered. A related BGC was identified in the marine‐derived Roussoella sp. DLM33 and confirmed by heterologous expression. These studies demonstrate that the biosynthetic pathways of terrein and related (polychlorinated) congeners diverge after oxidative decarboxylation of the lactone precursor that is catalyzed by a conserved FMO and further indicate that early dehydration of the side chain is an essential step

A Family of Nonribosomal Peptides Modulate Collective Behavior in Pseudovibrio Bacteria Isolated from Marine Sponges**

Although swarming motility and biofilms are opposed collective behaviors, both contribute to bacterial survival and host colonization. Pseudovibrio bacteria have attracted attention because they are part of the microbiome of healthy marine sponges. Two-thirds of Pseudovibrio genomes contain a member of a nonribosomal peptide synthetase-polyketide synthase gene cluster family, which is also found sporadically in Pseudomonas pathogens of insects and plants. After developing reverse genetics for Pseudovibrio, we isolated heptapeptides with an ureido linkage and related nonadepsipeptides we termed pseudovibriamides A and B, respectively. A combination of genetics and imaging mass spectrometry experiments showed heptapetides were excreted, promoting motility and reducing biofilm formation. In contrast to lipopeptides widely known to affect motility/biofilms, pseudovibriamides are not surfactants. Our results expand current knowledge on metabolites mediating bacterial collective behavior

A importância da promoção do uso racional de medicamentos realizada pelo Projeto de Assistência Farmacêutica Estudantil

The group of Proper Use of Medicines, inserted into the Student Pharmaceutical Care Project (PAFE), from the college of Pharmaceutical Sciences of UNESP – Araraquara, has been conducting orientation activities and promoting the Rational Use of Drugs (URM) since 1998. This work aims to emphasize the importance of the promotion of URM made by the students, taking into account the practice of self-medication for a large part of the population, and other issues related to improper treatment. The results obtained through questionnaires during the XVII Week of Pharmaceutical Care Student (SAFE), which show that self-medication is performed in parallel with the use of prescripted drugs.O estande de Uso Correto de Medicamentos, inserido no Projeto de Atenção Farmacêutica Estudantil (PAFE) da Faculdade de Ciências Farmacêuticas da Unesp – Araraquara, tem realizado atividades de orientação e promoção do Uso Racional de Medicamentos (URM) desde 1999. Este trabalho visa enfatizar a importância da promoção do URM feita pelos estudantes, levando em consideração a prática da automedicação por uma grande parte da população brasileira, e outras questões relacionadas à terapêutica inadequada. São utilizados os resultados obtidos através de questionários aplicados durante a XVII Semana de Atenção Farmacêutica Estudantil (SAFE), que revelam que a prática da automedicação é realizada paralelamente ao uso de medicamentos prescritos