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GenX (hexafluoropropylene oxide dimer acid) belongs to the group of per- and poly-fluoroalkyl substance (PFAS) compounds introduced to replace perfluorooctanoic acid (PFOA), which has been phased out in industrial and consumer product formulations. While GenX has been investigated in lab animals, there is limited information available regarding its uptake and translocation in wheat and rice. This study reports on a greenhouse experiment in which wheat and rice grown under flooded and non-flooded conditions were exposed to two GenX concentrations in the soil (0.4 mg kg-1 and 2 mg kg-1). GenX was analysed in the soil, porewater and shoots using targeted liquid chromatography-tandem mass spectroscopy (LC-MS/MS) analysis. Extractable organic fluorine (EOF) was determined using high-resolution continuum source graphite furnace molecular absorption spectrometry (HR-GFMAS) instrument. Results showed that different species took up different amounts of GenX. The GenX concentration in rice shoots was found to be 2.34 (± 0.45) and 4.11 (± 0.87) µg g-1 under flooded and non-flooded conditions, respectively, at a low exposure level. At high exposure, the GenX concentrations in flooded and non-flooded rice shoots increased threefold to 10.4 (± 0.41) and 13.4 (± 0.72) µg g-1, respectively. Wheat shoots showed similar concentrations and increases between low- and high-level exposure. The translocation factor was significantly higher (P = 0.013) in non-flooded rice compared to flooded rice. The GenX bioaccumulation behaviours under the same culture conditions (e.g. temperature, humidity, light, same GenX concentration in the soil) were significantly different in non-flooded and flooded rice (P < 0.001). Non-flooded rice plants displayed a higher level of GenX bioaccumulation than flooded ones. Following exposure to GenX, flooded rice plants showed a reduction in biomass (25%) compared to the control plants (P < 0.014). Our findings indicate that GenX is a bioaccumulative compound, the presence of which likely inhibits the growth of plants.
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Oryza , Poluentes do Solo , Animais , Solo , Triticum , Cromatografia Líquida , Espectrometria de Massas em Tandem , Poluentes do Solo/análiseRESUMO
Genome mining and metabolomics have become valuable tools in natural products research to evaluate and identify potential new chemistry from bacteria. In the search for new compounds from the deep-sea organism, Pseudomonas sp. SST3, from the South Shetland Trough, Antarctica, a co-cultivation with a second deep-sea Pseudomonas zhaodongensis SST2, was undertaken to isolate pseudomonassin, a ribosomally synthesised and post-translationally modified peptide (RiPP) that belongs to a class of RiPP called lasso peptides. Pseudomonassin was identified using a genome-mining approach and isolated by means of mass spectrometric guided isolation. Extensive metabolomics analysis of the co-cultivation of Pseudomonas sp. SST3 and P. zhaodongensis SST2, Pseudomonas sp. SST3 and Escherichia coli, and P. zhaodongensis SST2 and E. coli were performed using principal component analysis (PCA) and orthogonal projections to latent structures discriminant analysis (OPLS-DA), which revealed potential new metabolites in the outlier regions of the co-cultivation, with other metabolites identified previously from other species of Pseudomonas. The sequence of pseudomonassin was completely deduced using high collision dissociation tandem mass spectrometry (HCD-MS/MS). Preliminary studies on its activity against the pathogenic P. aeruginosa and its biofilm formation have been assessed and produced a minimum inhibitory concentration (MIC) of 63 µg/mL and 28 µg/mL, respectively.
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BACKGROUND: Following a well-balanced diet ensures that a person gets all the essential elements for health sustenance. However, in the United Kingdom an increasing proportion of people are transiting to become vegans who exclude animal-based products in their diets. Consequently, people may have a deficit of essential elements such as iodine which is not present in most plant-based meals, additionally iodide fortified table salt is not commonly used in the UK. Without iodine people consuming a vegan diet risk developing iodine deficiency and diseases like goiter. METHODS: The objective of this study is to determine the difference in iodine content and iodine speciation between plant-based and dairy products. More than 100 market samples of plant-based and dairy milk products were collected in Scotland, UK. RESULTS: Iodine concentrations in dairy milk is ten times higher compared to plant-based milks. Similar differences were also apparent for butter, yogurt and cheese. A total of 20% of plant-based milk products were fortified with iodine, however these products had lower iodine concentrations compare to the equivalent dairy products. In this study we calculated that people with average diet have an iodine intake of 226 + /- 103 µg day-1 from dairy products which satisfies the WHO recommended intake of adults and 90% of the recommend intake for pregnant and breast-feeding women. A diet from substituted dairy products gives only 21.8 µg day-1 for the respective WHO guideline intake values, which accounts only 15% of the iodine intake for adults and 9% for pregnant and lactating women. Iodine fortified diet could increase the iodine intake to 55% or 33% of the WHO recommended daily intake respectively. CONCLUSION: Plant-based dairy consumers are encouraged to use iodine fortified dairy products or use of iodized salt in the UK for home cooking, otherwise there are at risk to get iodine deficient.
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Iodo , Desnutrição , Gravidez , Animais , Feminino , Lactação , Laticínios , Leite/química , Dieta , Iodo/análise , Estado NutricionalRESUMO
Marine biofouling is the undesired accumulation of organic molecules, microorganisms, macroalgae, marine invertebrates, and their by-products on submerged surfaces. It is a serious challenge for marine vessels and the oil, gas, and renewable energy industries, as biofouling can cause economic losses for these industries. Natural products have been an abundant source of therapeutics since the start of civilisation. Their use as novel anti-fouling agents is a promising approach for replacing currently used, harmful anti-fouling agents. Anthraquinones (AQs) have been used for centuries in the food, pharmaceutical, cosmetics, and paint industries. Citreorosein and emodin are typical additives used in the anti-fouling paint industry to help improve the global problem of biofouling. This study is based on our previous study, in which we presented the promising activity of structurally related anthraquinone compounds against biofilm-forming marine bacteria. To help uncover the anti-fouling potential of other AQ-related structures, 2194 compounds from the COCONUT natural products database were analysed. Molecular docking analysis was performed to assess the binding strength of these compounds to the LuxP protein in Vibrio carchariae. The LuxP protein is a vital binding protein responsible for the movements of autoinducers within the quorum sensing system; hence, interrupting the process at an early stage could be an effective strategy. Seventy-six AQ structures were found to be highly docked, and eight of these structures were used in structure-based pharmacophore modelling, resulting in six unique pharmacophore features.
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Incrustação Biológica , Produtos Biológicos , Incrustação Biológica/prevenção & controle , Simulação de Acoplamento Molecular , Biofilmes , Produtos Biológicos/farmacologia , Antraquinonas/farmacologiaRESUMO
In this study, we explored a fungal strain UIAU-3F identified as Aspergillus fumigatus isolated from soil samples collected from the River Oyun in Kwara State, Nigeria. In order to explore its chemical diversity, the fungal strain UIAU-3F was cultured in three different fermentation media, which resulted in different chemical profiles, evidenced by LC-ESI-MS-based metabolomics and multivariate analysis. The methanolic extract afforded two known compounds, fumitremorgin C (1) and pseurotin D (2). The in vitro antiparasitic assays of 1 against Trypanosoma cruzi and Plasmodium falciparum showed moderate activity with IC50 values of 9.6 µM and 2.3 µM, respectively, while 2 displayed IC50 values > 50 µM. Molecular docking analysis was performed on major protein targets to better understand the potential mechanism of the antitrypanosomal and antiplasmodial activities of the two known compounds.
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Cowpox is caused by a DNA virus known as the cowpox virus (CPXV) belonging to the Orthopoxvirus genus in the family Poxviridae. Cowpox is a zoonotic disease with the broadest host range among the known poxviruses. The natural reservoir hosts of CPXV are wild rodents. Recently, the cases of orthopoxviral infections have been increasing worldwide, and cowpox is considered the most common orthopoxviral infection in Europe. Cowpox is often a self-limiting disease, although cidofovir or anti-vaccinia gammaglobulin can be used in severe and disseminated cases of human cowpox. In this computational study, a molecular docking analysis of thymine- and arabinofuranosyl-thymine-related structures (1-21) on two cowpox-encoded proteins was performed with respect to the cidofovir standard and a 3D ligand-based pharmacophore model was generated. Three chemical structures (PubChem IDs: 123370001, 154137224, and 90413364) were identified as potential candidates for anti-cowpox agents. Further studies combining in vitro and in silico molecular dynamics simulations to test the stability of these promising compounds could effectively improve the future design of cowpox virus inhibitors, as molecular docking studies are not sufficient to consider a ligand a potential drug.
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Vírus da Varíola Bovina , Varíola Bovina , Animais , Humanos , Vírus da Varíola Bovina/genética , Vírus da Varíola Bovina/metabolismo , Timina/metabolismo , Cidofovir/farmacologia , Ligantes , Simulação de Acoplamento Molecular , RoedoresRESUMO
The Nigerian Niger-Delta crude oil exploration often results in spills that affect indigenous medicinal plant biodiversity, likely changing the phytochemical profile of surviving species, their bioactivity or toxicity. In crude oil-rich Kokori and crude oil-free Abraka, classic examples of indigenous plants occupying the medicine-food interface include Vernonia amygdalina (VAL) and Ocimum gratissimum leaves (OGL). These plants are frequently utilised during pregnancy and in anaemia. To date, no scientific investigation has been reported on the potential changes to the phytochemical or bioactivity of the study plants. To discuss the similarities and dissimilarities in antisickling bioactivity and phytochemicals in VAL and OGL collected from Kokori (VAL-KK and OGL-KK) and Abraka (VAL-AB and OGL-AB), in silico, in vitro and comparative UPLC-QTOF-MS analysis was performed. Nine unique compounds were identified in OGL-KK, which have never been reported in the literature, while differences in antisickling potentials were observed in VAL-KK, OGL-KK and, VAL-AB, OGL-AB. Our findings show that VAL-AB and OGL-AB are richer and more diverse in phytochemicals and displayed a slightly higher antisickling activity than VAL-KK and OGL-KK. Ligand-based pharmacophore modelling was performed to understand the potential compounds better; this study may provide a basis for explaining the effect of crude oil spills on secondary metabolites and a reference for further research.
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Ocimum , Petróleo , Plantas Medicinais , Vernonia , Ocimum/química , Vernonia/química , Folhas de Planta , Extratos Vegetais/farmacologiaRESUMO
Monkeypox is caused by a DNA virus known as the monkeypox virus (MPXV) belonging to the Orthopoxvirus genus of the Poxviridae family. Monkeypox is a zoonotic disease where the primary significant hosts are rodents and non-human primates. There is an increasing global incidence with a 2022 outbreak that has spread to Europe in the middle of the COVID-19 pandemic. The new outbreak has novel, previously undiscovered mutations and variants. Currently, the US Food and Drug Administration (FDA) approved poxvirus treatment involving the use of tecovirimat. However, there has otherwise been limited research interest in monkeypox. Mitoxantrone (MXN), an anthracycline derivative, an FDA-approved therapeutic for treating cancer and multiple sclerosis, was previously reported to exhibit antiviral activity against the vaccinia virus and monkeypox virus. In this study, virtual screening, molecular docking analysis, and pharmacophore ligand-based modelling were employed on anthracene structures (1-13) closely related to MXN to explore the potential repurposing of multiple compounds from the PubChem library. Four chemical structures (2), (7), (10) and (12) show a predicted high binding potential to suppress viral replication.
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COVID-19 , Mpox , Animais , Humanos , Monkeypox virus , Mpox/diagnóstico , Mpox/tratamento farmacológico , Simulação de Acoplamento Molecular , Mitoxantrona/farmacologia , Reposicionamento de Medicamentos , Pandemias , Receptores de Droga , Primatas , RoedoresRESUMO
Bacteria belonging to the phylum Actinobacteria are a very good source of antibiotics, and indeed dominate the current clinical antibiotic space. This paper reports Mutactimycin AP, a new compound belonging to an anthracycline-type family of antibiotics, isolated from a Saccharothrix sp. This actinobacterial strain was isolated from the rhizosphere of lupine plants growing in the extreme hyper-arid Atacama Desert. Structural characterization was carried out using electrospray ionization-mass spectrometry (ESI-MS) and NMR spectroscopy in combination with molecular modelling. The compound was tested against the ESKAPE pathogens, where it showed activity against MRSA and five strains associated with bovine mastitis, where it showed activity against Enterococcus pseudoavium and Staphylycoccus Aureus subsp. Aureus.
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Actinobacteria , Actinomycetales , Bovinos , Animais , Feminino , Actinobacteria/química , Microbiologia do Solo , Bactérias , Antibacterianos/farmacologia , Clima DesérticoRESUMO
Natural products derived from marine sponges have exhibited bioactivity and, in some cases, serve as potent quorum sensing inhibitory agents that prevent biofilm formation and attenuate virulence factor expression by pathogenic microorganisms. In this study, the inhibitory activity of the psammaplin-type compounds, psammaplin A (1) and bisaprasin (2), isolated from the marine sponge, Aplysinellarhax, are evaluated in quorum sensing inhibitory assays based on the Pseudomonas aeruginosa PAO1 lasB-gfp(ASV) and rhlA-gfp(ASV) biosensor strains. The results indicate that psammaplin A (1) showed moderate inhibition on lasB-gfp expression, but significantly inhibited the QS-gene promoter, rhlA-gfp, with IC50 values at 14.02 µM and 4.99 µM, respectively. In contrast, bisaprasin (2) displayed significant florescence inhibition in both biosensors, PAO1 lasB-gfp and rhlA-gfp, with IC50 values at 3.53 µM and 2.41 µM, respectively. Preliminary analysis suggested the importance of the bromotyrosine and oxime functionalities for QSI activity in these molecules. In addition, psammaplin A and bisaprasin downregulated elastase expression as determined by the standard enzymatic elastase assay, although greater reduction in elastase production was observed with 1 at 50 µM and 100 µM. Furthermore, the study revealed that bisaprasin (2) reduced biofilm formation in P. aeruginosa.
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Pseudomonas aeruginosa , Percepção de Quorum , Antibacterianos/farmacologia , Proteínas de Bactérias/metabolismo , Biofilmes , Dissulfetos , Elastase Pancreática , Pseudomonas aeruginosa/fisiologia , Tirosina/análogos & derivados , Fatores de Virulência/metabolismoRESUMO
Vernonia amygdalina (VA) and Ocimum gratissimum (OG) are among the most frequently consumed vegetables in Kokori and Abraka communities of Delta State, Nigeria. However, the continuous crude oil exploration and spillages in Kokori may threaten their safety for use as food and medicine. Twelve samples of VA and OG obtained from crude oil-rich and crude oil-free communities were comparatively analysed for proximate composition, heavy metals, and cytotoxicity. Data obtained were subjected to various multivariate statistical techniques, including principal component analysis (PCA), biplot, and analysis of variance (ANOVA), to investigate the correlations between the vegetables from the different communities and the effect of crude oil exploration and spill on plant biomass. Results obtained indicate a significant difference (p < 0.05) in the proximate composition of VA and OG and higher heavy metal content for VA from the crude oil-spill Kokori. Two VA collections from Kokori were exceptionally toxic to cellular crustaceans.
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Five new phenolic siderophores 1-5 were isolated from the organic extract of a culture broth in a modified SGG medium of Pseudomonas sp. UIAU-6B, obtained from sediments collected from the Oyun river in North Central Nigeria. The structure of the new compounds, pseudomonin A-C (1-3) and pseudomobactin A and B (4 and 5) isolated alongside two known compounds, pseudomonine (6) and salicylic acid (7), were elucidated based on high-resolution mass spectrometry, 1D and 2D NMR analyses. The absolute configuration of the threonine residue in compounds 1-5 was determined by Marfey analysis. The antimicrobial evaluation of compound 4 exhibited the most potent activity against vancomycin-sensitive Enterococcus faecium VS144754, followed by 3 and 5, with MIC values ranging from 8 to 32 µg/mL. Compounds 2 and 3 exhibited moderate activity against Mycobacterium tuberculosis H37Rv, with MIC values of 7.8 and 15.6 µg/mL, respectively. Plausible biosynthetic hypotheses toward the new compounds 1-5 were proposed.
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The fatty acid (FA) compositions of ten seaweeds representative of Chlorophyta, Rhodophyta, and Ochrophyta from Kuwait in the Arabian Gulf region were determined and are discussed in the context of their potential nutritional perspectives for seaweed valorization. All the seaweeds had higher saturated fatty acid (SFA) and lower monounsaturated (MUFA) and polyunsaturated fatty acid (PUFA) contents than those typical of tropical environments. Palmitic, myristic, stearic, oleic, linoleic, α-linolenic, and stearidonic acids were the major FAs detected. Arachidonic, eicosapentaenoic, and docosahexaenoic acids were detected in minor amounts. Conserved fatty acid patterns revealed phylogenetic relationships among phyla, classes, and orders matching the molecular phylogenies at higher taxonomic ranks. Hierarchical clustering analyses clearly segregated different seaweeds (except Codium papillatum and Iyengaria stellata) into distinct groups based on their FA signatures. All but one species (Chondria sp.) had health-beneficial n6/n3 PUFAs (0.33:1-2.94:1) and atherogenic (0.80-2.52) and thrombogenic indices (0.61-5.17). However, low PUFA/SFA contents in most of the species (except Ulva spp.) may limit their utilization in the formulation of PUFA-rich functional foods. Ulva spp. had substantially high PUFAs with PUFA/SFA > 0.4, n6/n3 (0.33-0.66) and atherogenic (0.80-1.15) and thrombogenic indices (0.49-0.72), providing substantial potential for their utilization in food and feed applications.
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Epicotripeptin (1), a new cyclic tripeptide along with four known cyclic dipeptides (2-5) and one acetamide derivative (6) were isolated from seagrass-associated endophytic fungus Epicoccum nigrum M13 recovered from the Red Sea. Additionally, two new compounds, cyclodidepsipeptide phragamide A (7) and trioxobutanamide derivative phragamide B (8), together with eight known compounds (9-16), were isolated from plant-derived endophyte Alternaria alternata 13A collected from a saline lake of Wadi El Natrun depression in the Sahara Desert. The structures of the isolated compounds were determined based on the 1D and 2D NMR spectroscopic data, HRESIMS data, and a comparison with the reported literature. The absolute configurations of 1 and 7 were established by advanced Marfey's and Mosher's ester analyses. The antimicrobial screening indicated that seven of the tested compounds exhibited considerable (MIC range of 2.5-5 µg/mL) to moderate (10-20 µg/mL) antibacterial effect against the tested Gram-positive strains and moderate to weak (10-30 µg/mL) antibacterial effect against Gram-negative strains. Most of the compounds exhibited weak or no activity against the tested Gram-negative strains. On the other hand, four of the tested compounds showed considerable antibiofilm effects against biofilm forming Gram-positive and Gram-negative strains.
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Alternaria/metabolismo , Antibacterianos/farmacologia , Ascomicetos/metabolismo , Biofilmes/efeitos dos fármacos , Bactérias Aeróbias Gram-Negativas/efeitos dos fármacos , Bactérias Gram-Positivas/efeitos dos fármacos , Antibacterianos/isolamento & purificação , Biofilmes/crescimento & desenvolvimento , Fermentação , Bactérias Aeróbias Gram-Negativas/crescimento & desenvolvimento , Bactérias Gram-Positivas/crescimento & desenvolvimento , Testes de Sensibilidade Microbiana , Estrutura Molecular , Relação Estrutura-AtividadeRESUMO
Dehydroalanine (Dha) and dehydrobutyrine (Dhb) display considerable flexibility in a variety of chemical and biological reactions. Natural products containing Dha and/or Dhb residues are often found to display diverse biological activities. While the (Z) geometry is predominant in nature, only a handful of metabolites containing (E)-Dhb have been found thus far. Here we report discovery of a new antimicrobial peptide, albopeptide, through NMR analysis and chemical synthesis, which contains two contiguous unsaturated residues, Dha-(E)-Dhb. It displays narrow-spectrum activity against vancomycin-resistant Enterococcusâ faecium. In-vitro biochemical assays show that albopeptide originates from a noncanonical NRPS pathway featuring dehydration processes and catalysed by unusual condensation domains. Finally, we provide evidence of the occurrence of a previously untapped group of short unsaturated peptides in the bacterial kingdom, suggesting an important biological function in bacteria.
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Antibacterianos/biossíntese , Peptídeos Catiônicos Antimicrobianos/química , Alanina/análogos & derivados , Alanina/química , Aminobutiratos/química , Antibacterianos/química , Antibacterianos/farmacologia , Peptídeos Catiônicos Antimicrobianos/biossíntese , Peptídeos Catiônicos Antimicrobianos/farmacologia , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Avaliação Pré-Clínica de Medicamentos , Farmacorresistência Bacteriana/efeitos dos fármacos , Enterococcus faecium/efeitos dos fármacos , Família Multigênica , Ressonância Magnética Nuclear Biomolecular , Biossíntese de Peptídeos Independentes de Ácido Nucleico , Peptídeo Sintases/genética , Peptídeo Sintases/metabolismo , Estereoisomerismo , Streptomyces/enzimologia , Streptomyces/metabolismoRESUMO
Siderophores are iron-chelating compounds that aid iron uptake, one of the key strategies for microorganisms to carve out ecological niches in microbially diverse environments. Desferrioxamines are the principal siderophores produced by Streptomyces spp. Their biosynthesis has been well studied and as a consequence, the chemical potential of the pathway continues to expand. With all of this in mind, our study aimed to explore extremotolerant and lupine rhizosphere-derived Streptomyces sp. S29 for its potential antifungal capabilities. Cocultivation of isolate S29 was carried out with Aspergillus niger and Botrytis cinerea, both costly fungal phytopathogens in the wine industry, to simulate their interaction within the rhizosphere. The results indicate that not only is Streptomyces sp. S29 extraordinary at producing hydroxamate siderophores but uses siderophore production as a means to 'starve' the fungi of iron. High resolution LC-MS/MS followed by GNPS molecular networking was used to observe the datasets for desferrioxamines and guided structure elucidation of new desferrioxamine analogues. Comparing the new chemistry, using tools like molecular networking and MS2LDA, with the known biosynthesis, we show that the chemical potential of the desferrioxamine pathway has further room for exploration.
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Desferroxamina/metabolismo , Ferro/metabolismo , Lupinus/microbiologia , Rizosfera , Streptomyces/metabolismo , Antifúngicos/química , Antifúngicos/farmacologia , Cromatografia Líquida , Desferroxamina/química , Desferroxamina/farmacologia , Redes e Vias Metabólicas , Espectrometria de Massas em TandemRESUMO
Marine endophytic fungi from under-explored locations are a promising source for the discovery of new bioactivities. Different endophytic fungi were isolated from plants and marine organisms collected from Wadi El-Natrun saline lakes and the Red Sea near Hurghada, Egypt. The isolated strains were grown on three different media, and their ethyl acetate crude extracts were evaluated for their antimicrobial activity against a panel of pathogenic bacteria and fungi as well as their antioxidant properties. Results showed that most of the 32 fungal isolates initially obtained possessed antimicrobial and antioxidant activities. The most potent antimicrobial extracts were applied to three different cellulose containing fabrics to add new multifunctional properties such as ultraviolet protection and antimicrobial functionality. For textile safety, the toxicity profile of the selected fungal extract was evaluated on human fibroblasts. The 21 strains displaying bioactivity were identified on molecular basis and selected for chemical screening and dereplication, which was carried out by analysis of the MS/MS data using the Global Natural Products Social Molecular Networking (GNPS) platform. The obtained molecular network revealed molecular families of compounds commonly produced by fungal strains, and in combination with manual dereplication, further previously reported metabolites were identified as well as potentially new derivatives.
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Lasso peptides are a diverse class of ribosomally synthesized and post-translationally modified peptides (RiPPs). Their proteolytic and thermal stability alongside their growing potential as therapeutics has increased attention to these antimicrobial peptides. With the advent of genome mining, the discovery of RiPPs allows for the accurate prediction of putatively encoded structures, however, MSn experiments only provide partial sequence confirmation, therefore 2D NMR experiments are necessary for characterisation. Multiple MS/MS techniques were applied to two structurally characterized lasso peptides, huascopeptin and leepeptin, and one uncharacterized lasso peptide, citrulassin C, which was not isolable in sufficient quantity for NMR analysis. We have shown that MS2 can be used to elucidate the full amino acid sequences previously predicted with genome mining for this compound class. HCD was able to open the macrocycles and fragment the newly opened linear peptides, confirming the complete amino acid sequences of the characterised lasso peptides. In addition, to determine if this technique could be applied at the earliest stages of the isolation process, we targeted a lasso peptide found by genome mining, citrulassin C, and were able to fully elucidate the amino acid sequence using only MS2 from a semi-crude extract of Streptomyces huasconensis HST28T.
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Espectrometria de Massas/métodos , Peptídeos Cíclicos/genética , Análise de Sequência de Proteína/métodos , Sequência de Aminoácidos , Peptídeos Cíclicos/químicaRESUMO
An amendment to this paper has been published and can be accessed via a link at the top of the paper.