Mass Spectrometry and Metabolomics—New Approaches for Helminth Biochemical Studies

Metabolomics, the study of the endogenously synthesized small molecules repertoire (nonproteinaceous), is of great relevance for establishing a wide view of cell physiology at specific moments, linking metabolic profiles to phenotypes and genotypes. To better understand biological systems, such as helminths life cycle, helminthic infection, and host-parasite interaction, metabolomics studies are crucial. For that, mass spectrometry-based metabolomics is the most popular strategy. Nontargeted metabolomics allows researchers to profile entire metabolomes present in cells, tissues, biofluids, or even samples as complex as stools. Through different mass spectrometric techniques, it is possible to unveil chemical markers for helminths, such as Schistosoma mansoni (a trematode) and Ascaris lumbricoides (a nematode), in addition to study mechanisms of action for different drugs, which targets parasites. Therefore, mass spectrometry allows designing biochemical pathways that may clarify the processes of parasite life cycle, helminthic infection, and host-parasite interaction, providing targets to further interference for parasite control or even infection treatment

Catalase vs Peroxidase Activity of a Manganese(II) Compound: Identification of a Mn(III)-(μ-O)2-Mn(IV) Reaction Intermediate by Electrospray Ionization Mass Spectrometry and Electron Paramagnetic Resonance Spectroscopy

Herein, we report reactivity studies of the mononuclear water-soluble complex [Mn(II)(HPClNOL)(η1-NO3)(η2-NO3)] 1, where HPClNOL ) 1-(bis-pyridin-2-ylmethyl-amino)-3-chloropropan-2-ol, toward peroxides (H2O2 and tertbutylhydroperoxide). Both the catalase (in aqueous solution) and peroxidase (in CH3CN) activities of 1 were evaluated using a range of techniques including electronic absorption spectroscopy, volumetry (kinetic studies), pH monitoring during H2O2 disproportionation, electron paramagnetic resonance (EPR), electrospray ionization mass spectrometry in the positive ion mode [ESI(+)-MS], and gas chromatography (GC). Electrochemical studies showed that 1 can be oxidized to Mn(III) and Mn(IV). The catalase-like activity of 1 was evaluated with and without pH control. The results show that the pH decreases when the reaction is performed in unbuffered media. Furthermore, the activity of 1 is greater in buffered than in unbuffered media, demonstrating that pH influences the activity of 1 toward H2O2. For the reaction of 1 with H2O2, EPR and ESI(+)-MS have led to the identification of the intermediate [Mn(III)Mn(IV)(μ- O)2(PClNOL)2]+. The peroxidase activity of 1 was also evaluated by monitoring cyclohexane oxidation, using H2O2 or tert-butylhydroperoxide as the terminal oxidants. Low yields (<7%) were obtained for H2O2, probably because it competes with 1 for the catalase-like activity. In contrast, using tert-butylhydroperoxide, up to 29% of cyclohexane conversion was obtained. A mechanistic model for the catalase activity of 1 that incorporates the observed lag phase in O2 production, the pH variation, and the formation of a Mn(III)-(μ-O)2-Mn(IV) intermediate is proposed

Coenzyme Q10 or Creatine Counteract Pravastatin-Induced Liver Redox Changes in Hypercholesterolemic Mice

Statins are the preferred therapy to treat hypercholesterolemia. Their main action consists of inhibiting the cholesterol biosynthesis pathway. Previous studies report mitochondrial oxidative stress and membrane permeability transition (MPT) of several experimental models submitted to diverse statins treatments. The aim of the present study was to investigate whether chronic treatment with the hydrophilic pravastatin induces hepatotoxicity in LDL receptor knockout mice (LDLr-/-), a model for human familial hypercholesterolemia. We evaluated respiration and reactive oxygen production rates, cyclosporine-A sensitive mitochondrial calcium release, antioxidant enzyme activities in liver mitochondria or homogenates obtained from LDLr-/- mice treated with pravastatin for 3 months. We observed that pravastatin induced higher H2O2 production rate (40%), decreased activity of aconitase (28%), a superoxide-sensitive Krebs cycle enzyme, and increased susceptibility to Ca2+-induced MPT (32%) in liver mitochondria. Among several antioxidant enzymes, only glucose-6-phosphate dehydrogenase (G6PD) activity was increased (44%) in the liver of treated mice. Reduced glutathione content and reduced to oxidized glutathione ratio were increased in livers of pravastatin treated mice (1.5- and 2-fold, respectively). The presence of oxidized lipid species were detected in pravastatin group but protein oxidation markers (carbonyl and SH- groups) were not altered. Diet supplementation with the antioxidants CoQ10 or creatine fully reversed all pravastatin effects (reduced H2O2 generation, susceptibility to MPT and normalized aconitase and G6PD activity). Taken together, these results suggest that 1- pravastatin induces liver mitochondrial redox imbalance that may explain the hepatic side effects reported in a small number of patients, and 2- the co-treatment with safe antioxidants neutralize these side effects

Green and roasted arabica coffees differentiated by ripeness, process and cup quality via electrospray ionization mass spectrometry fingerprinting

Direct infusion electrospray ionization mass spectrometry in both the negative ESI(-)-MS and positive ESI(+)-MS ion modes are investigated to differentiate green and roasted Arabica coffees with different stages of ripeness (green, ripe and overripe), post-harvesting process (dry, wet and semi-wet) and coffees with different cup qualities. In the ESI(-)-MS of green coffees, ions from deprotonated fatty acids and chlorogenic acids are the most important for ripeness discrimination. In the ESI(+)-MS, maturity is differentiated by ions from protonated caffeine, chlorogenic acids and K+ adducts of fatty acids. To differentiate between post-harvesting process in both ionization modes, ions from fatty acids, chlorogenic acids, sugars and carboxylic acids generated in the fermentation process are the most representative. Roasted Arabica coffees are also well discriminated: in the ESI(-)-MS, ions from chlorogenic acids and short-chain organic acids derived from sugars are important. In the ESI(+)-MS, discrimination are mainly performed by low m/z ions such as protonated pyridine and alkylpiridines formed via trigonelline degradation. Both ESI(+)-MS and ESI(-)-MS are able to differentiate cup quality for Arabica roasted coffees and the ions used to perform discrimination are the same ones described in ripeness and post-harvesting processes.A habilidade da técnica de espectrometria de massas com infusão direta e ionização por eletronebulização (IES-EM), nos modos de íons positivos e negativos, foi avaliada na diferenciação de cafés Arábica verdes e torrados e com diferentes estágios de amadurecimento (verde, maduro e passado), processo pós-colheita (seco, úmido e semi-úmido) e cafés classificados por prova de xícara. No modo negativo, a análise dos cafés verdes mostrou que os íons correspondentes aos ácidos graxos e ácidos clorogênicos desprotonados são os mais importantes para a discriminação da maturidade. No modo positivo, a maturidade é diferenciada através de íons correspondentes a cafeína, ácidos clorogênicos protonados e adutos de K+ de ácidos graxos. Na diferenciação da pós-colheita, em ambos os modos de ionização, são mais importantes os íons correspondentes aos ácidos graxos, ácidos clorogênicos, açúcares e ácidos carboxílicos formados da fermentação. Cafés Arábica torrados também são discriminados com eficiência. No modo negativo, são importantes os íons correspondentes aos ácidos clorogênicos e ácidos orgânicos de cadeia curta, derivados de açúcares. No modo positivo, a discriminação é realizada por íons de baixa m/z tais como piridina e alquil piridinas protonadas, formadas através da degradação da trigonelina. Ambos os IES(+)-EM e IES(-)-EM são capazes de discriminar diferentes cafés Arábica torrados classificados por prova de xícara e os íons que permitem esta diferenciação são os mesmos descritos para a maturidade e processos pós-colheita.313321Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES

ATLANTIC EPIPHYTES: a data set of vascular and non-vascular epiphyte plants and lichens from the Atlantic Forest

Epiphytes are hyper-diverse and one of the frequently undervalued life forms in plant surveys and biodiversity inventories. Epiphytes of the Atlantic Forest, one of the most endangered ecosystems in the world, have high endemism and radiated recently in the Pliocene. We aimed to (1) compile an extensive Atlantic Forest data set on vascular, non-vascular plants (including hemiepiphytes), and lichen epiphyte species occurrence and abundance; (2) describe the epiphyte distribution in the Atlantic Forest, in order to indicate future sampling efforts. Our work presents the first epiphyte data set with information on abundance and occurrence of epiphyte phorophyte species. All data compiled here come from three main sources provided by the authors: published sources (comprising peer-reviewed articles, books, and theses), unpublished data, and herbarium data. We compiled a data set composed of 2,095 species, from 89,270 holo/hemiepiphyte records, in the Atlantic Forest of Brazil, Argentina, Paraguay, and Uruguay, recorded from 1824 to early 2018. Most of the records were from qualitative data (occurrence only, 88%), well distributed throughout the Atlantic Forest. For quantitative records, the most common sampling method was individual trees (71%), followed by plot sampling (19%), and transect sampling (10%). Angiosperms (81%) were the most frequently registered group, and Bromeliaceae and Orchidaceae were the families with the greatest number of records (27,272 and 21,945, respectively). Ferns and Lycophytes presented fewer records than Angiosperms, and Polypodiaceae were the most recorded family, and more concentrated in the Southern and Southeastern regions. Data on non-vascular plants and lichens were scarce, with a few disjunct records concentrated in the Northeastern region of the Atlantic Forest. For all non-vascular plant records, Lejeuneaceae, a family of liverworts, was the most recorded family. We hope that our effort to organize scattered epiphyte data help advance the knowledge of epiphyte ecology, as well as our understanding of macroecological and biogeographical patterns in the Atlantic Forest. No copyright restrictions are associated with the data set. Please cite this Ecology Data Paper if the data are used in publication and teaching events. © 2019 The Authors. Ecology © 2019 The Ecological Society of Americ

Ácido fólico em leite e bebida láctea enriquecidos: estudo da vida-de-prateleira

O ácido fólico é uma vitamina, que desempenha funções importantes, o que justifica seu uso nos processos de enriquecimento dos alimentos. O objetivo deste trabalho foi avaliar a estabilidade do ácido fólico, utilizado no enriquecimento, durante a vida-de-prateleira de amostras dos seguintes produtos: leites em pó, leites esterilizados e bebidas lácteas achocolatadas prontas para o consumo. A determinação desta vitamina foi feita por cromatografia líquida de alta eficiência. O ácido fólico foi extraído com solução básica e a limpeza do extrato foi realizada com ácido tricloroacético concentrado seguida de filtração. No processo cromatográfico utilizou-se coluna C18, sistema de eluição por gradiente, sendo a fase móvel composta por tampão acetato e acetonitrila. A detecção foi feita na região do ultravioleta, a 290nm, e a quantificação por padronização externa. Nas amostras de leites em pó durante o período de estocagem de um ano ocorreram perdas acentuadas desta vitamina (60% em média), enquanto nas de leites esterilizados e bebidas lácteas achocolatadas as perdas foram, em média, de 25 e 29%, respectivamente, durante os quatro meses de estocagem