NP3 MS Workflow: An Open-Source Software System to Empower Natural Product-Based Drug Discovery Using Untargeted Metabolomics.

Natural products (or specialized metabolites) are historically the main source of new drugs. However, the current drug discovery pipelines require miniaturization and speeds that are incompatible with traditional natural product research methods, especially in the early stages of the research. This article introduces the NP3 MS Workflow, a robust open-source software system for liquid chromatography-tandem mass spectrometry (LC-MS/MS) untargeted metabolomic data processing and analysis, designed to rank bioactive natural products directly from complex mixtures of compounds, such as bioactive biota samples. NP3 MS Workflow allows minimal user intervention as well as customization of each step of LC-MS/MS data processing, with diagnostic statistics to allow interpretation and optimization of LC-MS/MS data processing by the user. NP3 MS Workflow adds improved computing of the MS2 spectra in an LC-MS/MS data set and provides tools for automatic [M + H]+ ion deconvolution using fragmentation rules; chemical structural annotation against MS2 databases; and relative quantification of the precursor ions for bioactivity correlation scoring. The software will be presented with case studies and comparisons with equivalent tools currently available. NP3 MS Workflow shows a robust and useful approach to select bioactive natural products from complex mixtures, improving the set of tools available for untargeted metabolomics. It can be easily integrated into natural product-based drug-discovery pipelines and to other fields of research at the interface of chemistry and biology.

Machine learning-driven serum metabolomics: insights into the transition to early-stage heart failure with preserved ejection fraction

Introduction: Metabolomics has emerged as a powerful tool in providing readouts of early disease states before clinical manifestation. Here we used the predictive power of Unsupervised Hierarchical Clustering Analysis (UHCA) and Automated Machine Learning (AutoML) algorithms to identify serum metabolic panels in a population at risk of developing HFpEF. Methods: We studied 215 subjects staged as non-HF, pre-HFpEF and early-stage HFpEF(es-HFpEF). We evaluated clinical, laboratory, echocardiographic, and NMR-based metabolomics of blood serum data. UHCA and AutoML were used to explore metabolic fingerprints potentially related to clinical features or HFpEF. We used Metabolite Set Enrichment Analysis to explore biochemical pathways. Results: The UHCA identified three major patients (P) and two metabolites (M) clusters (Figure). The P clusters were associated with HFpEF stages, cardiac remodeling, diastolic dysfunction, and sex (Pearson Chi-square, p < 0.05) and M clusters with glycine and serine metabolism and urea cycle pathways (FDR-adjusted p-value < 0.002). Considering non-HFpEF and es-HFpEF groups, AUROC mean for feature subset combinations was 0.897 and the highest AUROC (0.995) combined metabolites, clinical, laboratory and echo features. Of the 64 models trained that included metabolites as input, serine (25), uridine (17), 2-oxoglutarate (14), citrate (14), 2-aminobutyrate (13) and taurine (13) were observed more frequently with feature importance value greater than zero. The metabolites with higher sum values of feature importance were serine (0.173), uridine (0.131), 2-aminobutyrate (0.123), choline (0.098) and dimethylamine (0.087). Conclusions: This study revealed characteristic metabolite profiles in the sera of patients at risk of developing HFpEF. These metabolite panels can add information for classificatory algorithms development and contribute to the understanding of HFpEF pathophysiology.

Flavored Amazonic pirarucu (Arapaima giga) waste flour (salted and sweet) for inclusion in food products.

This study aimed to develop flavored flours (salty and sweet) from Amazonic pirarucu waste (Arapaima gigas), include them in extruded snacks, and evaluate the nutritional, physicochemical, microbiological, and acceptance characteristics of these products. A standard flour was elaborated with pirarucu carcass, which presented 54.42% of protein and 7.24% of lipids, and from this, flavored flours were elaborated (salty and sweet). The standard flour had higher levels of protein, calcium, and phosphorus; and the salted one had higher levels of lipids. The fatty acids present in greater quantities were oleic (average of 32.21%), linolenic (average of 20.74%), and palmitic (average of 17.81%). The flavored flours were better accepted than the standard flour, for all sensory attributes and purchase intention. The snacks with sweet flour, despite better results in the sensory attributes of color, aroma, and flavor, were the ones that presented the lowest content of protein and ash, when compared to those with inclusion of standard flour. It is concluded that the pirarucu waste can be used for producing flavored flours and extruded snacks, with the purpose of improved food products.

Production of Epoxyketone Peptide-Based Proteasome Inhibitors by Streptomyces sp. BRA-346: Regulation and Biosynthesis.

Streptomyces sp. BRA-346 is an Actinobacteria isolated from the Brazilian endemic tunicate Euherdmania sp. We have reported that this strain produces epoxyketone peptides, as dihydroeponemycin (DHE) and structurally related analogs. This cocktail of epoxyketone peptides inhibits the proteasome chymotrypsin-like activity and shows high cytotoxicity to glioma cells. However, low yields and poor reproducibility of epoxyketone peptides production by BRA-346 under laboratory cultivation have limited the isolation of epoxyketone peptides for additional studies. Here, we evaluated several cultivation methods using different culture media and chemical elicitors to increase the repertoire of peptide epoxyketone production by this bacterium. Furthermore, BRA-346 genome was sequenced, revealing its broad genetic potential, which is mostly hidden under laboratory conditions. By using specific growth conditions, we were able to evidence different classes of secondary metabolites produced by BRA-346. In addition, by combining genome mining with untargeted metabolomics, we could link the metabolites produced by BRA-346 to its genetic capacity and potential regulators. A single biosynthetic gene cluster (BGC) was related to the production of the target epoxyketone peptides by BRA-346. The candidate BGC displays conserved biosynthetic enzymes with the reported eponemycin (EPN) and TMC-86A (TMC) BGCs. The core of the putative epoxyketone peptide BGC (ORFs A-L), in which ORF A is a LuxR-like transcription factor, was cloned into a heterologous host. The recombinant organism was capable to produce TMC and EPN natural products, along with the biosynthetic intermediates DH-TMC and DHE, and additional congeners. A phylogenetic analysis of the epn/tmc BGC revealed related BGCs in public databases. Most of them carry a proteasome beta-subunit, however, lacking an assigned specialized metabolite. The retrieved BGCs also display a diversity of regulatory genes and TTA codons, indicating tight regulation of this BGC at the transcription and translational levels. These results demonstrate the plasticity of the epn/tmc BGC of BRA-346 in producing epoxyketone peptides and the feasibility of their production in a heterologous host. This work also highlights the capacity of BRA-346 to tightly regulate its secondary metabolism and shed light on how to awake silent gene clusters of Streptomyces sp. BRA-346 to allow the production of pharmacologically important biosynthetic products.

Guia prático para o manejo da dor / Guia prático para o manejo da dor

O estudo da dor e suas particularidades é de grande importância para o tratamento de diversas patologias e para a melhora na qualidade de vida dos pacientes. A maioria das disfunções orgânicas tem a dor como um ponto importante da sua manifestação. Dessa maneira, é justificável a elaboração de conteúdo para auxiliar os profissionais da saúde no entendimento e tratamento das principais causas de dores agudas e crônicas. Este livro foi elaborado com o objetivo de servir como um guia prático para o manejo da dor por profissionais e acadêmicos de Medicina. Engloba temas como conceitos e aspectos biopsicossociais da dor, além de questões mais complexas como a fisiologia da dor e o tratamento medicamentoso com o arsenal terapêutico existente. Finalmente, também trata dos diversos tipos de dor mais prevalentes e o conhecimento básico que envolve seu manejo.

Seasonal and spatial variations in limnological conditions of a floodplain lake (lake Catalão) connected to both the Solimões and Negro Rivers, Central Amazonia / Variações espaciais e temporais nas condições limnológicas de um lago de planície de inundação (lago Catalão) conectado aos rios Solimões e Negro, Amazônia Central

Lakes play an important role in biogeochemical, ecological and hydrological processes in the river-floodplain system. The aim of this study was to evaluate the dynamics of the limnological conditions of Catalمo Lake, an Amazon floodplain lake. Thus, some of the main limnological environment variables (O2, temperature, pH, nutrient, electrical conductivity) of the Catalمo Lake were analyzed under temporal and spacial scales. The study was conducted between November/2004 and August/2005. Sampling excursion were carried out every three months; one excursion for each of the four different hydrological periods (low water, rising water, high water and falling water). Sampling points were chosen so that it could be obtained a gradient of the distance from Negro River. Limnological profiles in Catalمo Lake showed generally acidic to slightly alcaline water, with low levels of dissolved oxygen and low concentrations of soluble reactive phosphorous. The Negro River seems to exert the main influence during the rising water period, while the Solimُes River is the principal controlling river during peak water. The Principal Component Analysis (PCA) grouped the seasonal collections by hydrological period, showing the formation of a north-south spatial gradient within the lake in relation to the limnological variables. Multivariate dispersion analysis based on distance-to-centroid method demonstrated an increase in similarity over the course of the hydrological cycle, as the lake was inundated in response to the flood pulse of the main river channels. However, the largest spatial homogeneity in the lake was observed in the epilimnion layer, during the falling water period. The daily analysis of variation indicated an oligomitic pattern during the years in which the lake was permanently connected to the Negro River...

Os lagos exercem um papel importante nos processos biogeoquímicos, ecológicos e hidrológicos no sistema rio-planície de inundação. O objetivo do presente estudo foi avaliar a dinâmica das condições limnológicas do lago Catalão, um lago da planície de inundação amazônica. Algumas das principais variáveis limnológicas (por exemplo, O2, temperatura, pH, condutividade elétrica, nutrientes) do lago Catalão foram analisadas em uma escala temporal e espacial. O presente estudo foi realizado no período de novembro de 2004 a agosto de 2005. As amostragens foram trimestrais, com um total de quatro coletas, uma em cada fase do ciclo hidrológico (Seca, Enchente, Cheia e Vazante). Os pontos de amostragem foram escolhidos de modo a obter um gradiente de distância em relação ao rio Negro. No geral, as águas do lago Catalão mostraram-se pouco oxigenadas, ácidas a levemente alcalinas e com baixas concentrações de Fósforo Solúvel Reativo (FSR). A Análise de Componentes Principais (ACP) agrupou as estações de coleta por fases do ciclo hidrológico e evidenciou a formação de um gradiente espacial na distribuição das variáveis limnológicas, que vai da região mais ao norte até mais ao sul do lago. A influência do rio Negro sobre as águas do lago parece ser maior na enchente e a do rio Solimões na cheia. A análise de dispersão multivariada com abordagem baseada na distância ao centróide evidenciou o aumento da similaridade espacial em função da inundação. Contudo, a maior homogeneidade espacial do lago foi registrada no epilímnio, na fase de vazante. Os resultados da análise de variação diária indicaram que o lago Catalão apresenta comportamento oligomítico nos anos em que permanece conectado permanentemente ao rio Negro. Apesar de receber grande aporte de águas pretas (rio Negro) e brancas (rio Solimões), as características físicas e químicas das águas do Catalão permitem classificá-lo como um sistema mais próximo dos lagos de várzea típicos do que daqueles de águas preta.

Hibernoma do couro cabeludo / Hibernoma of the hairy scalp

O hibernoma constitui-se em tumor benigno raro, derivado de gordura marrom que usualmente se manifesta como massa de crescimento lento e indolor. Este relato descreve a presença de hibernoma localizado no couro cabeludo em paciente adulta, feminino.

Hibernoma is a rare benign tumor of brown fat origin which usually manifests as a slowly growing, pain- less, soft-tissue mass. This case report describes the presence of hibernoma located on the scalp in an adult woman, female.