Chemical Composition, Repellent, and Phytotoxic Potentials of the Fractionated Resin Essential Oil from Araucaria heterophylla Growing in Tunisia.

The resin essential oil (REO) of the Tunisian Araucaria heterophylla trunk bark was investigated for its chemical composition. Then, it was evaluated for its insecticidal and allelopathic activities. The REO was obtained by hydrodistillation for 9 h (yield of 4.2 % w/w). Moreover, fractional hydrodistillation was carried out at 3-hour intervals, resulting in 3 fractions (R1-R3), to facilitate chemical identification and localization of the aforementioned biological activities. GC/MS analysis of the obtained samples allowed the identification of 25 compounds, representing between 91.2 and 96.3 % of their total constituents, which consisted predominantly of sesquiterpene hydrocarbons, oxygenated sesquiterpenes and diterpene hydrocarbons. α-Copaene (10.8 %), γ-muurolene (5.8 %), α-copaen-11-ol (7.8 %), spathulenol (10.5 %), 15-copaenol (8.2 %), ylangenal (10.3 %), dehydrosaussurea lactone (7.7 %), and sandaracopimaradiene (11.4 %) were identified as major compounds. The second part aimed to assess the impact of the A. heterophylla EO and its three fractions for their insecticidal and repellent activity against Tribolium castaneum (Herbst), a stored grain pest, of which a strong repellent activity was noted. In addition, the studied samples showed high phytotoxic effects against Lactuca sativa. The third fraction (R3) performed a total inhibitory potential on seed germination and seedling growth of the target plant. Furthermore, alongside this discovery, an estimation was conducted through molecular docking analysis. Wherein the main compounds of the studied samples were docked into the active pocket of protoporphyrinogen IX oxidase (PDB: 1SEZ), a key enzyme in chlorophyll biosynthesis. Thus, it is recommended to use the REO of A. heterophylla as a natural herbicide.

Bioactivity screening of pinhão (Araucaria Angustifolia (Bertol.) Kuntze) seed extracts: the inhibition of cholinesterases and α-amylases, and cytotoxic and anti-inflammatory activities.

The objective of this work was to determine the potential bioactive properties of extracts from bio-residues of pinhão (Araucaria angustifolia (Bertol.) Kuntze) seeds, namely the α-amylase and cholinesterase inhibition, cytotoxicity, and anti-inflammatory properties. The pinhão extracts evaluated were obtained from cooking water (CW) and as an ethanolic extract from residual pinhão seed shells (PS). Catechin was the major compound found in both extracts. The PS extract presented higher antioxidant levels and the better inhibition of human salivary and porcine pancreatic α-amylases when compared to the CW extract. Also, based on in vivo evaluations, the PS extract did not differ significantly from acarbose when compared to a control group. The most potent inhibitor of cholinesterases was the CW extract. No cytotoxicity toward normal cells was detected, and neither extract showed anti-inflammatory activity. The PS extract presented cytotoxic activity toward non-small-cell lung, cervical, hepatocellular and breast carcinoma cell lines. Overall, the results demonstrated the potential bioactivity of extracts obtained from pinhão bio-residues.

Oral and Topical Anti-Inflammatory and Antipyretic Potentialities of Araucaria bidiwillii Shoot Essential Oil and Its Nanoemulsion in Relation to Chemical Composition.

Different parts of Araucaria bidiwillii (bunya pin) trees, such as nuts, seeds, bark, and shoots, are widely used in cooking, tea, and traditional medicines around the world. The shoots essential oil (EO) has not yet been studied. Herein, the chemical profile of A. bidiwillii shoots EO (ABSEO) was created by GC-MS analysis. Additionally, the in vivo oral and topical anti-inflammatory effect against carrageenan-induced models, as well as antipyretic potentiality of ABSEO and its nanoemulsion were evaluated. Forty-three terpenoid components were identified and categorized as mono- (42.94%), sesqui- (31.66%), and diterpenes (23.74%). The main compounds of the ABSEO were beyerene (20.81%), α-pinene (16.21%), D-limonene (14.22%), germacrene D (6.69%), ß-humulene (4.14%), and sabinene (4.12%). The ABSEO and its nanoemulsion exhibited significant inflammation suppression in carrageenan-induced rat paw edema model, in both oral (50 and 100 mg/kg) and topical (5% in soyabean oil) routes, compared to the control and reference drugs groups. All the results demonstrated the significant inflammation reduction via the inflammatory cytokines (IL-1ß and IL8), nitrosative (NO), and prostaglandin E2 (PGE2) supported by the histopathological studies and immunohistochemical assessment of MMP-9 and NF-κß levels in paw tissues. Moreover, the oral administration of ABSEO and its nanoemulsion (50 and 100 mg/kg) exhibited antipyretic activity in rats, demonstrated by the inhibition of hyperthermia induced by intramuscular injection of brewer's yeast. These findings advised that the use of ABSEO and its nanoemulsion against numerous inflammatory and hyperthermia ailments that could be attributed to its active constituents.

Pinhão coat extract encapsulated in starch ultrafine fibers: Thermal, antioxidant, and antimicrobial properties and in vitro biological digestion.

This study aimed to produce soluble potato starch ultrafine fibers for the encapsulation of pinhão coat extract (PCE), evaluating their relative crystallinity (RC), thermal stability, antioxidant activity, antimicrobial activity against Escherichia coli and Staphylococcus aureus, as well as in vitro biological digestion. In the simulation of in vitro biological digestion, the phenolic compounds release profile was also evaluated. The ultrafine fibers were produced by electrospinning, based on a polymeric solution composed of soluble potato starch (50% w/v) and formic acid. Then, PCE was incorporated at various concentrations (0.5%, 1.0%, and 1.5%, w/w, dry basis). The endothermic event of free PCE was not observed in the ultrafine fibers, which suggests its encapsulation. The RC decreased according to the increase in PCE concentration in the ultrafine fibers. The PCE resisted thermal treatments when encapsulated into the ultrafine fibers (100 and 180°C), and the ultrafine fibers with 1% PCE presented the highest amount of preserved phenolic compounds. Regarding antioxidant activity, the free PCE presented 85% of DPPH inhibition and the ultrafine fibers had 18% inhibition, not differing among the PCE concentrations (p < 0.05). The free PCE and the ultrafine fibers with 0.5% PCE showed inhibitory effect against S. aureus and the ones with 1.5% PCE showed controlled release of phenolic compounds during the simulation of in vitro digestion. Starch ultrafine fibers showed potential to be applied in food industries due to their capacity of protecting phenolic compounds when submitted to high temperatures or gastrointestinal conditions. Nevertheless, their application depends on the end use of the product. PRACTICAL APPLICATION: The encapsulation of pinhão coat extract (PCE) in ultrafine starch fibers promotes greater preservation of phenolic compounds. Thus, it can be incorporated into different foods that are produced using the ultra-high temperature (UHT) process-at 135-145°C for 5 to 10 s, or some other equivalent time/temperature combination. Another possibility is the incorporation of ultrafine fibers in active packaging: compounds can migrate to food, improving sensory characteristics, increasing shelf life, preventing chemical and microbiological deterioration, and ensuring food safety.

Aerogels based on corn starch as carriers for pinhão coat extract (Araucaria angustifolia) rich in phenolic compounds for active packaging.

The objective of this study was to produce renewable aerogels from native and anionic corn starches loaded with pinhão coat extract (PCE) with water absorbent capacity, antioxidant activity and controlled release of phenolic compounds in a hydrophilic food simulant media. Starch aerogels were produced with different concentrations of PCE, 5 and 10%, and evaluated for FT-IR spectra, relative crystallinity, thermal properties, water absorption capacity (WAC), density, antioxidant activity and in vitro release. Thermal stability of the compounds was improved by the incorporation of PCE. The aerogels presented high WAC of 541 to 731% and low-density values of 0.03 g.cm-1. The highest inhibition of DPPH and ABTS radicals was presented to anionic starch aerogels with 10% PCE rendering 26% of inhibition of ABTS and 24% of DPPH. The maximum in vitro releases for native and anionic starch aerogels with 5% of PCE were 28.70 and 29.44%, respectively, and for aerogels with 10% of PCE they were 34.27 and 35.94%, respectively. The anionic starch aerogels had the highest amount of phenolic compounds released when compared to the native starch aerogels. The starch-based bioactive aerogels showed potential to be applied in food packaging as water absorbent and as a carrier of phenolic compounds.

Extraction, Purification, and Characterization of Polysaccharides of Araucaria heterophylla L and Prosopis chilensis L and Utilization of Polysaccharides in Nanocarrier Synthesis.

BACKGROUND: Plant gums consist of polysaccharides which can be used in the preparation of nanocarriers and provide a wide application in pharmaceutical applications including as drug delivery agents and the matrices for drug release. The objectives of the study were to collect plant gums from Araucaria heterophylla L and Prosopis chilensis L and to extract and characterize their polysaccharides. Then to utilize these plant gum-derived polysaccharides for the formulation of nanocarriers to use for drug loading and to examine their purpose in drug delivery in vitro. METHODS: Plant gum was collected, polysaccharide was extracted, purified, characterized using UV-Vis, FTIR, TGA and GCMS and subjected to various bioactive studies. The purified polysaccharide was used for making curcumin-loaded nanocarriers using STMP (sodium trimetaphosphate). Bioactivities were performed on the crude, purified and drug-loaded nanocarriers. These polysaccharide-based nanocarriers were characterized using UV-Vis spectrophotometer, FTIR, SEM, and AFM. Drug release kinetics were performed for the drug-loaded nanocarriers. RESULTS: The presence of glucose, xylose and sucrose was studied from the UV-Vis and GCMS analysis. Purified polysaccharides of both the plants showed antioxidant activity and also antibacterial activity against Bacillus sp. Purified polysaccharides were used for nanocarrier synthesis, where the size and shape of the nanocarriers were studied using SEM analysis and AFM analysis. The size of the drug-loaded nanocarriers was found to be around 200 nm. The curcumin-loaded nanocarriers were releasing curcumin slow and steady. CONCLUSION: The extracted pure polysaccharide of A. heterophylla and P. chilensis acted as good antioxidants and showed antibacterial activity against Bacillus sp. These polysaccharides were fabricated into curcumin-loaded nanocarriers whose size was below 200 nm. Both the drug-loaded nanocarriers synthesized using A. heterophylla and P. chilensis showed antibacterial activity with a steady drug release profile. Hence, these natural exudates can serve as biodegradable nanocarriers in drug delivery.

Assessment of the use of cooked Araucaria angustifolia seed coats extract as food for brine shrimp.

The use of vegetable waste and its screening for potential cytotoxicity is of utmost importance to ensure its safe use in the feed industry for fish and other animals. We evaluated aqueous and ethanolic extracts of cooked Araucaria angustifolia seed coats. The Stiasny index for the aqueous and ethanolic extracts was 2.87% ± 0.03% and 60.53% ± 4.79%, respectively. Condensed tannins were 11-fold higher in the ethanolic extract than the aqueous extract. The flavonoid and polyphenol contents were 1.7- and 1.8-fold higher in the ethanolic extract than in the aqueous extract, respectively. The 36 h EC50 for brine shrimp hatchability was 300.32 µg/mL for the aqueous extract, and 76.60 µg/mL for the ethanolic extract. The 24 h LC50 was 1405.96 µg/mL for the aqueous extract, and it was 356.32 µg/mL for the ethanolic extract. The aqueous extract was nontoxic to A. salina nauplii, and therefore, it can be used as a possible food additive in fish feed. The results also demonstrated that the different solvents used in the extraction affected the yield and the total phenolic, total flavonoid, and condensed tannin content. Further in vivo and cell line cytotoxicity testing is recommended to substantiate these findings.

Evaluation of volatile compounds, phenolic acids, antioxidant potential and DFT study of essential oils from different parts of Araucaria columnaris (G. Forst.) Hook. from India.

This study is aimed to investigate the aroma composition, phenolic acid, antioxidant activities and computational analysis of essential oils of five different parts of A. columnaris (G. Forst.) Hook. from India. The essential oils were obtained from the fresh root, stem, leaves, bark, and branch of A. columnaris by hydro-distillation using a Clevenger apparatus: the volatile composition was examined by GC-MS; the phenolic acid quantification by RP-HPLC-DAD and the estimation of antioxidant potential by TAC, FRAP, CUPRAC, MCA, DRSC, and NOSC assays. Total 136 chemical compounds i.e. root-60, stem-55, leaves-50, bark-55 and branch-42 were identified in essential oils by GC-MS and major constituents were ß-pinene, terpinolene, 2-nonanone, pinocarveol, trans-verbenol, dihydrocarvone, dodecane, thymol, isolongifolene, α-copaene, α-gurjunene, ß-caryophyllene, isoledene, γ-muurolene, γ-cadinene, etc. The five phenolic acid contents were determined by RP-HPLC-DAD with AR: 91-99%, LOD: 0.010269-0.119857 mg/g and LOQ: 0.039553-0.003388 mg/g. The Density Functional Theory (DFT) study was carried out on the main phytochemicals present in essential oils of A. columnaris in a manner to support the experimental results and give detailed of antioxidants and reactivity parameters. The results suggested that the essential oils of A. columnaris have high antioxidant potential due to the presence of phenolic acids and aroma compositions.

Updating embryonic ontogenesis in Araucaria angustifolia: from Burlingame (1915) to the present.

This study addresses gaps in our understanding of pre-fertilization and archegonia development and reinterprets embryonic ontogenesis from Burlingame (Bot Gaz 59:1-39, 1915) to the present based on timescale and structural features allowing us to determine functionally and developmentally accurate terminology for all these stages in A. angustifolia. Different from previous reports, only after pollination, pre-fertilization tissue development occurs (0-13 months after pollination (MAP)) and gives rise to a mature megagametophyte. During all this period, pollen is in a dormant state at the microphyla, and pollen tube germination in nucellus tissue is only observed at the stage of archegonia formation (13 MAP) and not at the free nuclei stage as reported before. For the first time, 14 months after pollination, a fertilization window was indicated, and at 15 MAP, the polyzygotic polyembryony from different archegonia was also seen. After that, subordinated proembryo regression occurs and at least three embryonic developmental stages of dominant embryo were characterized: proembryogenic, early embryogenic, and late embryogenic (15-23 MAP). Along these stages, histochemical and ultrastructural analyses suggest the occurrence of cell death in suspensor and in cap cells of dominant embryo that was not previously reported. The differentiation of meristems, procambium, pith, and cortex tissues in late embryogenic stage was detailed. The morphohistological characterization of pre-fertilization and embryonic stages, together with the timescale of megastrobili development, warranted a referential map of female reproductive structure in this species.

Utilization of gum polysaccharide of Araucaria heterophylla and Azadirachta indica for encapsulation of cyfluthrin loaded super paramagnetic iron oxide nanoparticles for mosquito larvicidal activity.

Surface engineering of super paramagnetic iron oxide nanoparticles (SPIONs) favor the tagging of any molecule or compound onto it, encapsulating them with a biopolymer make them biocompatible and favor slow release of loaded molecules. Recovery of SPIONs is easier as they obey to external magnetic field. In this study, SPIONS were used for mosquito larvicidal activity after surface engineered with oleic acid to favor the tagging of Cyfluthrin (mosquito larvicidal agent), it was then encapsulated with gum polysaccharide derived from Azadirachta indica and Araucaria heterophylla. Every stage of coreshell formation was microscopically and spectroscopically characterized. The coreshell SPIONs produced using Azadirachta indica and Araucaria heterophylla gum derived polysaccharide encapsulation were found to be the size around 80 nm. Thus, prepared coreshell SPIONs was subjected for mosquito larvicidal activity against Culex sp. The coreshell SPIONs was efficiently killing the mosquito larva and its impact was studied by percentage mortality studies.

Araucaria angustifolia (Bertol.) Kuntze extract as a source of phenolic compounds in TPS/PBAT active films.

There is growing interest in the development of biodegradable packaging materials containing natural antioxidant extracts. In this sense, the use of extracts obtained from agro-industrial byproducts has proved to be a sustainable alternative. In this study, Pinhão extract, a byproduct of Pinhão (Araucaria angustifolia (Bertol.) Kuntze) seed consumption, was characterized by HPLC-DAD-ESI/MSn, demonstrating the presence of eight phenolic compounds, (+)-catechin and (-)-epicatechin being the most abundant molecules. TPS/PBAT films containing Pinhão extract were produced by blown extrusion and their properties (tensile properties, thermal characteristics and microstructure) were evaluated in order to determine the effect of the presence of extracts. Results suggested that the interaction between the phenolic compounds of the extract and the polymeric matrix caused the reduction in the crystallinity degree, and an increase in the starch glass transition temperature. The presence of Araucaria angustifolia (Bertol.) Kuntze extract significantly (p < 0.05) affected the color and opacity of the film. Regarding water vapor permeation, no significant difference (p > 0.05) was detected. However, the water solubility and the contact angle with water (polar solvent) and diiodomethane (non-polar solvent) significantly changed due to the extract addition. Moreover, the Pinhão extract conferred significant antioxidant capacity to the TPS/PBAT films as determined by DPPH, suggesting that this material can be applied as an active packaging material.

Purification, characterization and utilization of polysaccharide of Araucaria heterophylla gum for the synthesis of curcumin loaded nanocarrier.

In this study, gum of Araucaria heterophylla was collected. The collected gum was subjected for extraction of polysaccharide using solvent extraction system. Thus, extracted polysaccharide was further purified using solvent method and was characterized using UV-Vis spectroscopy, Phenol sulfuric acid assay, FTIR, TGA, TLC and GC-MS. The gum derived polysaccharide was found to have the following sugars Rhamnose, Allose, Glucosinolate, Threose, Idosan, Galactose and Arabinose. The extracted polysaccharide was tested for various in-vitro bioactive studies such as antibacterial activity, antioxidant activity and anticancer activity. The polysaccharide was found to have antioxidant and anticancer activity. Further, the polysaccharide was subjected for carboxymethylation to favor the nanocarrier synthesis, where it was chelated using Sodium Tri Meta Phosphate (STMP) to form nanocarriers. The nanocarriers so formed were loaded with curcumin and were characterized using FTIR, SEM, EDX and AFM. Both the loaded and unloaded nanocarriers were studied for its in-vitro cytotoxic effect against the MCF7 human breast cancer cell lines. The nanocarriers were found to deliver the drug efficiently against the cancer cell line used in this study.