Quali-Quantitative Characterization of Volatile and Non-Volatile Compounds in Protium heptaphyllum (Aubl.) Marchand Resin by GC-MS Validated Method, GC-FID and HPLC-HRMS2.

Protium heptaphyllum (Aubl.) Marchand (PH) trees are endemic to the tropical region of South America, mostly Brazil. Antibacterial, antinociceptive, anti-inflammatory, anxiolytic, antidepressant and anti-hyperlipidemic/anti-hypercholesterolemic effects were reported for its resinous exudate Protiumheptaphyllum resin (PHR). This work aims to provide a qualitative and quantitative consistent chemical profiling of the major constituents of this resin and two extracts enriched in acid (acidic triterpene concentrated extract, ATCE) and neutral triterpenes (α and ß-amyrin concentrated extract, AMCE). GC-MS/GC-FID was used for volatile terpene fraction, a validated GC-MS method was developed for quantification of neutral α and ß-amyrin and HPLC-APCI HRMS2 was used for acidic triterpenes analysis. The chemical investigation reported 29 molecules, including 14 volatile terpenes, 6 neutral triterpenes and 11 acid triterpenes. The most abundant compounds were α-amyrin (251.28 g kg-1, 123.98 g kg-1 and 556.82 g kg-1 in PHR, ATCE and AMCE, respectively), ß-amyrin (172.66 g kg-1, 95.39 g kg-1 and 385.58 g kg-1 in PHR, ATCE and AMCE, respectively), 3-oxo-tirucalla-7,24-dien-21-oic acid (80.64 g kg-1, 157.10 g kg-1 and 15.31 g kg-1 in PHR, ATCE and AMCE, respectively) and 3α-hydroxy-tirucalla-8,24-dien-21-oic acid (77.71 g kg-1, 130.40 g kg-1 and 11.64 g kg-1 in PHR, ATCE and AMCE, respectively). Results showed specific enrichment of acidic and neutral triterpenoids in the two respective extracts.

Bioactive Triterpenes of Protium heptaphyllum Gum Resin Extract Display Cholesterol-Lowering Potential.

Hypercholesterolemia is one of the major causes of cardiovascular disease, the risk of which is further increased if other forms of dyslipidemia occur. Current therapeutic strategies include changes in lifestyle coupled with drug administration. Statins represent the most common therapeutic approach, but they may be insufficient due to the onset of resistance mechanisms and side effects. Consequently, patients with mild hypercholesterolemia prefer the use of food supplements since these are perceived to be safer. Here, we investigate the phytochemical profile and cholesterol-lowering potential of Protium heptaphyllum gum resin extract (PHE). Chemical characterization via HPLC-APCI-HRMS2 and GC-FID/MS identified 13 compounds mainly belonging to ursane, oleanane, and tirucallane groups. Studies on human hepatocytes have revealed how PHE is able to reduce cholesterol production and regulate the expression of proteins involved in its metabolism. (HMGCR, PCSK9, LDLR, FXR, IDOL, and PPAR). Moreover, measuring the inhibitory activity of PHE against HMGR, moderate inhibition was recorded. Finally, molecular docking studies identified acidic tetra- and pentacyclic triterpenoids as the main compounds responsible for this action. In conclusion, our study demonstrates how PHE may be a useful alternative to contrast hypercholesterolemia, highlighting its potential as a sustainable multitarget natural extract for the nutraceutical industry that is rapidly gaining acceptance as a source of health-promoting compounds.

Differentiation of black and white pitch (Burseraceae) oleoresins: A mass spectrometry-based chemoethnotaxonomic study.

ETHNOPHARMACOLOGICAL RELEVANCE: In the quilombola communities of the municipality of Oriximiná (Pará State, Brazil), Protium spp. (Burseraceae) oleoresins are distinguished in black and white pitch. White pitch oleoresins may be superior to black pitch in terms of quality, but the criteria used for their differentiation are fairly subjective. AIM OF THE STUDY: This study was designed to provide a scientific rationale for the traditional differentiation of black and white pitch oleoresins based on their non-volatile fraction. MATERIALS AND METHODS: Black and white pitch oleoresin samples collected in quilombola territories in Oriximiná were analysed by GC-EI-MS and UPLC-APCI-MS. The feasibility of EI and APCI mass spectrometry-based pattern recognition methods PLS-DA and Random Forest Analysis (RFA) for black and white pitch oleoresins differentiation was demonstrated. RESULTS: The UPLC-APCI-MS method allowed the separation of 43 triterpenoids. Assessment of the triterpenoid fingerprints by GC-EI-MS led to the tentative identification of ursa-9(11),12-dien-3-ol as a potential marker for black pitch oleoresins. PLS-DA and RFA applied to the APCI-MS and EI-MS data gave good models for black and white pitch oleoresins classification. The most important ions for the classifications of black pitch oleoresins by APCI-MS/PLS-DA and APCI-MS/RFA likely represented triterpenoid acids. CONCLUSIONS: The triterpenoid pattern differs between black and white pitch oleoresins. The characteristic presence of ursa-9(11),12-dien-3-ol and triterpenoids acids in black pitch oleoresins, along with other field observations, suggest that black pitch oleoresins are actually aged white pitch oleoresins.

Spatial distribution pattern and diameter structure of Protium Burm. f. in Iratapuru river sustainable development reserve, Amapá, Brazil / Padrão de distribuição espacial e estrutura diamétrica de Protium Burm. f. na reserva de desenvolvimento sustentável do Rio Iratapuru, Amapá, Brasil

The present work aims to determine the spatial distribution pattern of Protium Burm. f and the diametric structure of these species in a dense tropical submontane rainforest along the Sustainable Development Reserve (SDR) of the Iratapuru river. The area of study was divided into three plateau compartments called Mane Preto Sierra, Banco Grande Sierra and Alaska Sierra. In each plateau compartment it was settled a single area transect with 1,000 x 20 m, where DAP ≥ 10 cm of the entire Protium population within the transects was measured. The data was processed in the specific software Mata Nativa 2. There were inventoried Protium tenuifolium, Protium altsoni, Protium polybotryum subsp. blackii and Protium paniculatum var. riedelianum. Most individuals concentrated in smaller diametric classes, decreasing progressively to a lesser extent in larger classes. It can be concluded that the population has medium size and its vertical structure is characterized by the presence of three strata, however, its phytocenoses is defined by two clear constants: the first groups individuals of up to 15 m of height and forms the uniform and dominant stratum; the second is composed by emerging up to 35 m high and whose diameter structure shows a population in dynamic balance.

Este trabalho tem como objetivo determinar o padrão de distribuição espacial de Protium Burm. f e estrutura diamétrica dessas espécies em uma floresta tropical densa submontana ao longo da reserva de desenvolvimento sustentável (RDS) do rio Iratapuru. A área estudada foi dividida em três compartimentos de planaltos denominados Serra Mane Preto, Serra Banco Grande e Serra Alaska. Em cada compartimento de planalto instalou-se transecto de área única com 1.000 x 20 m, onde foi mensurado o DAP ≥ 10 cm de toda população de Protium dentro dos transectos. Os dados foram processados em softwares específicos Mata Nativa 2. Foram inventariados Protium tenuifolium, Protium altsonii, Protium polybotryum subsp. blackii e Protium paniculatum var. riedelianum. A maior parte dos indivíduos concentrou-se nas menores classes diamétricas, diminuindo progressivamente até atingir menor proporção nas maiores classes. Pode-se concluir que a população apresenta porte mediano, sua estrutura vertical caracteriza-se pela presença de três estratos; no entanto, sua fitocenose define-se por dois contínuos bem evidentes; o primeiro agrupa indivíduos com até 15 m de altura e forma o estrato uniforme e dominante; o segundo é composto por emergentes com até 35 m de altura e que a estrutura diamétrica evidencia uma população em equilíbrio dinâmico.

Chemical Composition, Enantiomeric Distribution, and Antifungal Activity of the Oleoresin Essential Oil of Protium amazonicum from Ecuador.

Background:Protium species (Burseraceae) have been used in the treatment of various diseases and conditions such as ulcers and wounds. Methods: The essential oil from the oleoresin of Protium amazonicum was obtained by hydrodistillation and analyzed by GC-MS, GC-FID, and chiral GC-MS. P. amazonicum oleoresin oil was screened for antifungal activity against Candida albicans, Aspergillus niger, and Cryptococcus neoformans. Results: A total of 54 components representing 99.6% of the composition were identified in the oil. The essential oil was dominated by δ-3-carene (47.9%) with lesser quantities of other monoterpenoids α-pinene (4.0%), p-cymene (4.1%), limonene (5.1%), α-terpineol (5.5%) and p-cymen-8-ol (4.8%). Chiral GC-MS revealed most of the monoterpenoids to have a majority of levo enantiomers present with the exceptions of limonene and α-terpineol, which showed a dextro majority. P. amazonicum oleoresin oil showed promising activity against Cryptococcus neoformans, with MIC = 156 µg/mL. Conclusions: This account is the first reporting of both the chemical composition and enantiomeric distribution of the oleoresin essential oil of P. amazonicum from Ecuador. The oil was dominated by (-)-δ-3-carene, and this compound, along with other monoterpenoids, likely accounts for the observed antifungal activity of the oil.

Report on the Malungo expedition to the Erepecuru river, Oriximiná, Brazil. Part I: is there a difference between black and white breu?

ABSTRACT Species belonging to Burseraceae produce an oleoresin known in the north of Brazil as breu. They comprise an essential oil with a complex composition, and are used in Amazonia for smoking the environment, to caulk boats and for medicinal purposes. Depending on its organoleptic characteristics and on the breu-producing species, they are called white or black breu. In this work, we provide data about the breu-producing species occurring in the quilombola region of the Erepecuru river, the chemical composition, and whether it is possible to differentiate them based on their chemical composition and/or botanical identification. Aerial samples from breu trees and oleoresins were collected from 10 different individuals at 6 different sites on the Erepecuru river under the guidance of the quilombolas. Essential oils were extracted by hydrodistillation and characterized by GC–MS. From the analysis, 126 different substances were identified, with a large quantitative and qualitative variation. To better understand the chemical variations within the samples and to sort the variation into the categories of white or black breu as identified by the quilombola, we sorted the oil samples into five different sets according to their major compounds (A: δ-3-carene; B: p-cymene; C: γ-cadinene/p-cymene; D: limonene, β-phellandrene/α-terpineol; E: α-pinene/limonene). Essential oils from samples of white breu had the highest concentration of α-pinene, while a similarity in chemical composition could not be established for the black breu samples (sets A, B and C). Furthermore, a chemical similarity between a black breu (Protium heptaphyllum (Aubl.) Marchand) and a white breu (Protium decandrum (Aubl.) Marchand) sample was evidenced. In conclusion, it is difficult to establish definitions for white and black breu based on chemical, botanical or regional names. This designation is more cultural and regional than scientific and is based on the oleoresin production volume, its color aspect and scent.

ElemeNT: a computational tool for detecting core promoter elements.

Core promoter elements play a pivotal role in the transcriptional output, yet they are often detected manually within sequences of interest. Here, we present 2 contributions to the detection and curation of core promoter elements within given sequences. First, the Elements Navigation Tool (ElemeNT) is a user-friendly web-based, interactive tool for prediction and display of putative core promoter elements and their biologically-relevant combinations. Second, the CORE database summarizes ElemeNT-predicted core promoter elements near CAGE and RNA-seq-defined Drosophila melanogaster transcription start sites (TSSs). ElemeNT's predictions are based on biologically-functional core promoter elements, and can be used to infer core promoter compositions. ElemeNT does not assume prior knowledge of the actual TSS position, and can therefore assist in annotation of any given sequence. These resources, freely accessible at http://lifefaculty.biu.ac.il/gershon-tamar/index.php/resources, facilitate the identification of core promoter elements as active contributors to gene expression.

TRF2: TRansForming the view of general transcription factors.

Transcriptional regulation is pivotal for development and differentiation of organisms. Transcription of eukaryotic protein-coding genes by RNA polymerase II (Pol II) initiates at the core promoter. Core promoters, which encompass the transcription start site, may contain functional core promoter elements, such as the TATA box, initiator, TCT and downstream core promoter element. TRF2 (TATA-box-binding protein-related factor 2) does not bind TATA box-containing promoters. Rather, it is recruited to core promoters via sequences other than the TATA box. We review the recent findings implicating TRF2 as a basal transcription factor in the regulation of diverse biological processes and specialized transcriptional programs.

Comparative overview of RNA polymerase II and III transcription cycles, with focus on RNA polymerase III termination and reinitiation.

In eukaryotes, RNA polymerase (RNAP) III transcribes hundreds of genes for tRNAs and 5S rRNA, among others, which share similar promoters and stable transcription initiation complexes (TIC), which support rapid RNAP III recycling. In contrast, RNAP II transcribes a large number of genes with highly variable promoters and interacting factors, which exert fine regulatory control over TIC lability and modifications of RNAP II at different transitional points in the transcription cycle. We review data that illustrate a relatively smooth continuity of RNAP III initiation-elongation-termination and reinitiation toward its function to produce high levels of tRNAs and other RNAs that support growth and development.

Mel Amargo de Breu (Protium sp., Burseraceae)

A sample of bitter a honey collected in the Mucajaí county (Roraima, Brazil) forest of Apiaú, was analysed. The pollen analysis revealed a predominance of Protium pollen, comprising 60,9% of the total of pollen grains.

Foi analisada uma amostra de mel amargo procedente do Município de Mucajaí (Roraima, Brasil), Floresta de Apiaú. A análise polínica constatou a dominância de pólen de breu (Protium sp.) compreendendo 60,9% do total de grãos de pólen.