Phytochemicals Determination, and Antioxidant, Antimicrobial, Anti-Inflammatory and Anticancer Activities of Blackberry Fruits.

A comprehensive characterization of the phytochemicals present in a blackberry fruit extract by HPLC-TOF-MS has been carried out. The main compounds in the extract were ursane-type terpenoids which, along with phenolic compounds, may be responsible for the bioactivity of the extract. In vitro antioxidant capacity was assessed through Folin-Ciocalteu (31.05 ± 4.9 mg GAE/g d.w.), FRAP (637.8 ± 3.2 µmol Fe2+/g d.w.), DPPH (IC50 97.1 ± 2.4 µg d.w./mL) and TEAC (576.6 ± 8.3 µmol TE/g d.w.) assays. Furthermore, the extract exerted remarkable effects on in vitro cellular antioxidant activity in HUVEC cells at a concentration of 5 mg/mL. Antimicrobial activity of the extract was also tested. Most sensible microorganisms were Gram-positive bacteria, such as E. faecalis, B. cereus and Gram-negative E. coli (MBC of 12.5 mg/mL). IC50 values against colon tumoral cells HT-29 (4.9 ± 0.2 mg/mL), T-84 (5.9 ± 0.3 mg/mL) and SW-837 (5.9 ± 0.2 mg/mL) were also obtained. Furthermore, blackberry extract demonstrated anti-inflammatory activity inhibiting the secretion of pro-inflammatory IL-8 cytokines in two cellular models (HT-29 and T-84) in a concentration-dependent manner. These results support that blackberry fruits are an interesting source of bioactive compounds that may be useful in the prevention and treatment of different diseases, mainly related to oxidative stress.

Terpenic Constituents of Essential Oils with Larvicidal Activity against Aedes Aegypti: A QSAR and Docking Molecular Study.

Aedes aegypti is a vector for the arbovirus responsible for yellow fever, Zika and Chikungunya virus. Essential oils and their constituents are known for their larvicidal properties and are strong candidates for mosquito control. This work aimed to develop a quantitative structure-activity study and molecular screening for the search and design of new larvicidal agents. Twenty-five monoterpenes with previously evaluated larvicidal activity were built and optimized using computational tools. QSAR models were constructed through genetic algorithms from the larvicidal activity and the calculation of theoretical descriptors for each molecule. Docking studies on acetylcholinesterase (AChE) and sterol carrier protein (SCP-2) were also carried out. Results demonstrate that the epoxide groups in the structure of terpenes hinder larvicidal activity, while lipophilicity plays an important role in enhancing biological activity. Larvicidal activity correlates with the interaction of the sterol-carrier protein. Of the 25 compounds evaluated, carvacrol showed the highest larvicidal activity with an LC50 of 8.8 µg/mL. The information included in this work contributes to describing the molecular, topological, and quantum mechanical properties related to the larvicidal activity of monoterpenes and their derivatives.

Terpene-Containing Analogues of Glitazars as Potential Therapeutic Agents for Metabolic Syndrome.

Metabolic syndrome is a complex of abnormalities involving impaired glucose and lipid metabolism, which needs effective pharmacotherapy. One way to reduce lipid and glucose levels associated with this pathology is the simultaneous activation of nuclear PPAR-alpha and gamma. For this purpose, we synthesized a number of potential agonists based on the pharmacophore fragment of glitazars with the inclusion of mono- or diterpenic moiety in the molecular structure. The study of their pharmacological activity in mice with obesity and type 2 diabetes mellitus (C57Bl/6Ay) revealed one substance that was capable of reducing the triglyceride levels in the liver and adipose tissue of mice by enhancing their catabolism and expressing a hypoglycemic effect connected with the sensitization of mice tissue to insulin. It has also been shown to have no toxic effects on the liver.

Influence of genetic and interannual factors on bioactive compounds of olive pomace determined through a germplasm survey.

Olive mill wastes, generated in the extraction of virgin olive oil (VOO), are of important concern for the industry owing to the produced volume and polluting load, mainly associated with the presence of organic compounds. Among them, it is worth mentioning bioactive compounds, mainly phenols and triterpenes, which could be potentially isolated for further use in the cosmetic, pharmaceutical, or food industries. This research analyzed the olive pomace after extraction of VOO from fruits harvested of 43 international olive cultivars during three consecutive seasons. The cultivar was identified as the most determinant factor to explain the variability in the relative concentration of phenols and terpenic acids in the extracts. In addition, the characterization of olive pomace extracts allowed clustering cultivars according to the profile of bioactive compounds. Finally, we identified the components responsible for the observed discrimination that was explained according to biosynthetic metabolic pathways.

Chemical Composition and Assessment of Antimicrobial Activity of Lavender Essential Oil and Some By-Products.

The producers of essential oils from the Republic of Moldova care about the quality of their products and at the same time, try to capitalize on the waste from processing. The purpose of the present study was to analyze the chemical composition of lavender (Lavanda angustifolia L.) essential oil and some by-products derived from its production (residual water, residual herbs), as well as to assess their "in vitro" antimicrobial activity. The gas chromatography-mass spectrometry analysis of essential oils produced by seven industrial manufacturers led to the identification of 41 constituents that meant 96.80-99.79% of the total. The main constituents are monoterpenes (84.08-92.55%), followed by sesquiterpenes (3.30-13.45%), and some aliphatic compounds (1.42-3.90%). The high-performance liquid chromatography analysis allowed the quantification of known triterpenes, ursolic, and oleanolic acids, in freshly dried lavender plants and in the residual by-products after hydrodistillation of the essential oil. The lavender essential oil showed good antibacterial activity against Bacillus subtilis, Pseudomonas fluorescens, Xanthomonas campestris, Erwinia carotovora at 300 µg/mL concentration, and Erwinia amylovora, Candida utilis at 150 µg/mL concentration, respectively. Lavender plant material but also the residual water and ethanolic extracts from the solid waste residue showed high antimicrobial activity against Aspergillus niger, Alternaria alternata, Penicillium chrysogenum, Bacillus sp., and Pseudomonas aeroginosa strains, at 0.75-6.0 µg/mL, 0.08-0.125 µg/mL, and 0.05-4.0 µg/mL, respectively.

Lipase-Catalyzed Esterification of Geraniol and Citronellol for the Synthesis of Terpenic Esters.

This article describes the synthesis of terpenic esters derived from geraniol and citronellol (geranyl and citronellyl alkanoates) through esterification reactions catalyzed by the immobilized lipases from Thermomyces lanuginosus (Lipozyme TL IM®) and Candida antarctica (Novozym 435®). Geraniol was esterified with oleic, lauric, and stearic acids; and citronellol was esterified with oleic and stearic acids. For all the synthesized flavor esters, the best conditions were 35 °C, and the molar ratio between acid and alcohol was 1:1. Geranyl and citronellyl alkanoates reached yields between 80-100% within 4 h of reaction. For the synthesis of the citronellyl and geranyl oleate, higher yields were obtained in the absence of organic solvents. For the esters from lauric and stearic acids, using solvent was indispensable to improve the miscibility between the substrates. The reuse of Novozym 435® and Lipozyme TL IM® was performed for two more cycles after the first use, with yields higher than 60%. The results demonstrated the efficiency of the reaction catalyzed by these two commercial enzymes and the feasibility of the methodology for the production of synthetic flavor esters through enzymatic catalysis. The flavor esters synthesized were not described in the literature up to the date, giving this research an innovative feature.

Unveiling the lager beer volatile terpenic compounds.

Beer volatile terpenic compounds arise from a network of variables, namely from plant raw-materials metabolism, yeast metabolism, among others, and may suffer modifications during brewing. In order to increase the molecular understanding of beer volatile terpenic compounds, this work intends to perform a comprehensive characterization of these molecules on lager beer, using an advanced multidimensional chromatographic methodology. This research comprises the most detailed screening of lager beer terpenic compounds through the putative identification of 94 mono and sesquiterpenic compounds, distributed over 6 chemical families: alcohols, aldehydes, esters, ketones, hydrocarbons, and oxides. The terpenic profiles allowed the beer terpen-typing according to samples' category clustering: macro and micro-brewer beers. This study adds further insight to the lager beer terpenic volatile composition, and could be applied in large-scale studies in different contexts, namely for understanding the distinctive beer styles or for beer typing.

New Metabolites Isolated from a Laurencia obtusa Population Collected in Corsica.

The chemical investigation of an ethyl acetate extract (EtOAc) obtained from Laurencia obtusa, collected in Corsica, allowed for the identification of three new compounds (1, 2, and 4) and six known compounds. Compounds 1 to 4 were isolated and fully characterized by a detailed spectroscopic analysis. Compounds 1 and 2 are two C15-acetogenins sharing the same ring system: a tetrahydropyran linked by a methylene to a tetrahydrofuran ring. Compound 1 exhibits a bromoallene unit whereas compound 2 possesses an uncommon α-bromo-α,ß-unsaturated aldehyde terminal unit. Compound 4 is the first diterpene exhibiting a 19(4 → 3)abeo-labdane skeleton isolated from a Laurencia species. Isolation of concinndiol (compound 3) together with compound 4 suggests a common biosynthetic origin. Additionally, five known compounds, namely sagonenyne, laurene, α-bromocuparene, microcladallene A, and ß-snyderol were identified in chromatographic fractions by NMR analysis using a computerized method that was developed in our laboratory.

Adsorption of terpenic compounds onto organo-palygorskite.

Essential oils from aromatic plants are currently mentioned as suitable tools for excellent protection of stored grains from insect pest attacks. The present work aimed to study the processes of the synthesis insecticidal formulation with clay. The active terpenic compounds of essential oil of Eucalyptus globulus (Eg) were fixed in the palygorskite by adsorption process. Two sample types of palygorskites were used: raw and organo-palygorskite. The palygorskite clays were characterized by different physicochemical techniques including X-ray diffraction (XRD), Fourier transform infrared (FTIR) analyses, thermogravimetric analysis (TG), differential thermal analysis (DTA), Brunauer-Emmet-Teller (BET), and scanning electron microscope (SEM). Results reveal that the raw clay has a fibrous structure with impurities essential calcite. These structures and physicochemical properties of raw palygorskite and organo-palygorskite give it the potential of material adsorbent. Results show that the adsorption capacity strongly depends on affinity between terpenic compounds and organic cations rather than on interlayer distance of organo-palygorskite. The highest adsorption capacity of terpenic compounds is acquired with palygorskite interlaced by didodecyldimethylammonium bromide (DDDMA). These results validated the potential utility of the Paly-DDDMA as adsorbent fibrous clay for the retention of terpenic compounds in application of environmental preservation.

Unveiling elderflowers (Sambucus nigra L.) volatile terpenic and norisoprenoids profile: Effects of different postharvest conditions.

The volatile terpenic and norisoprenoids profile from elderflowers (Sambucus nigra L.) was established for two cultivars by multidimensional gas chromatography. From 47 monoterpenic, 13 sesquiterpenes and 5 norisoprenoids components, 38 are reported for the first time on elderflowers. Elderflower seasonality implies proper handling and storage conditions, for further processing, thus the impact of freezing, freeze-drying, air drying and vacuum packing, was evaluated on these potential aroma metabolites. The most suitable preservation methods, regarding the total metabolites content, were vacuum packing and freezing for intermediary storage times (24-32weeks) with a reported overall decrease of the volatile terpenic and norisoprenoids of up to 58.6%; and freezing, for longer period (52weeks), with a decrease of up to 47.4% (compared to fresh elderflowers). This study presents the most detailed terpenic and norisoprenoids elderflower profiling, and linalool oxides were proposed as markers for a more expedite assess to the impact of postharvest conditions.

Floral aroma improvement of Muscat spirits by packed column distillation with variable internal reflux.

The organoleptic quality of wine distillates depends on raw materials and the distillation process. Previous work has shown that rectification columns in batch distillation with fixed reflux rate are useful to obtain distillates or distillate fractions with enhanced organoleptic characteristics. This study explores variable reflux rate operating strategies to increase the levels of terpenic compounds in specific distillate fractions to emphasize its floral aroma. Based on chemical and sensory analyses, two distillate heart sub-fractions obtained with the best operating strategy found, were compared with a distillate obtained in a traditional alembic. Results have shown that a drastic reduction of the reflux rate at an early stage of the heart cut produced a distillate heart sub-fraction with a higher concentration of terpenic compounds and lower levels of negative aroma compounds. Therefore, this sub-fraction presented a much more noticeable floral aroma than the distillate obtained with a traditional alembic.

Metabolomic-Based Strategy for Fingerprinting of Sambucus nigra L. Berry Volatile Terpenoids and Norisoprenoids: Influence of Ripening and Cultivar.

The integration of plant metabolomics to support preharvest fruit development studies can provide important insights into the biochemical mechanisms involved and lately support producers on harvesting management. A metabolomic-based strategy for fingerprinting of volatile terpenoids and norisoprenoids from Sambucus nigra L. berries from three cultivars, through ripening, was established. From 42 monoterpenic, 20 sesquiterpenic, and 14 norisoprenoid compounds, 48 compounds are reported for the first time as S. nigra berries components. Chemometric tools revealed that ripening was the factor that influenced more the volatile fraction profile and physicochemical parameters (pH, TS, and TSS), followed by cultivar. For the unripe stages, a higher overall content of the studied metabolites was observed, which gradually decreased over the ripening stages, being consistent for the three cultivars. These trends were mainly ruled by limonene, p-cymene, aromadendrene, ß-caryophyllene, and dihydroedulan, which might therefore be used by producers as an additional simple decision making tool in conjunction with physicochemical parameters.

Natural products: Perspectives in the pharmacological treatment of gastrointestinal anisakiasis.

OBJECTIVE: To evaluate the activity of different natural products against L3 larvae of Anisakis type I. METHODS: Information on investigations into the activity of natural products against the L3 larvae of Anisakis simplex was collected from scientific journals, books, theses and reports via a library and electronic search (using Pubmed, Scopus, Medline, Web of Science and ScienceDirect). The search terms included: natural products, medicinal plants, essential oils, terpenic derivatives, Anisakis, antinematodal activity. RESULTS: In the literature reviewed numerous papers were found concerning the in vitro and in vivo activity against Anisakis type I of various natural products (plant extracts, essential oils and their major components). Analysis of the results showed that in vitro tests the Melaleuca alternifolia essential oil and the ar-turmerone isolated from Curcuma longa displayed the greatest activity. In vivo, the most active compound was perillaldehyde, the main component of Perilla frutescens essential oil. CONCLUSIONS: This study shows that some natural products exhibited promising antianisakis properties.

Quantitation of selected terpenoids and mercaptans in the dual-purpose hop varieties Amarillo, Citra, Hallertau Blanc, Mosaic, and Sorachi Ace.

Free terpenoids and both free and bound polyfunctional thiols were investigated in five selected dual-purpose hop cultivars. Surprisingly, the dual-purpose Sorachi Ace variety was found to contain higher amounts of farnesene (2101 mg/kg) than aromatic hops such as Saaz but only traces of 3-methylbutylisobutyrate, a compound that usually distinguishes all bitter varieties. All five cultivars investigated here showed an exceptional citrus-like potential explained by either monoterpenic alcohols or polyfunctional thiols. Among the monoterpenic alcohols, ß-citronellol at concentrations above 7 mg/kg distinguished Amarillo, Citra, Hallertau Blanc, Mosaic, and Sorachi Ace from Nelson Sauvin and Tomahawk, two previously investigated dual-purpose hops, while linalool (312 mg/kg) and geraniol (211 mg/kg) remained good discriminating compounds for Nelson Sauvin and Tomahawk, respectively. Regarding polyfunctional thiols, higher amounts of 3-sulfanylhexyl acetate (27 µg/kg) characterized the Citra variety. Free 4-sulfanyl-4-methylpentan-2-one proved discriminant for Sorachi Ace, while the bound form is predominant in Nelson Sauvin. On the other hand, an S-conjugate of 3-sulfanylhexan-1-ol was found in Sorachi Ace at levels not far from those previously reported for Cascade, although the free form was undetected here. Both free and bound grapefruit-like 3-sulfanyl-4-methylpentan-1-ol (never evidenced before the present work) emerged as discriminating compounds for the Hallertau Blanc variety. The apotryptophanase assay also allowed us to evidence for the first time an S-conjugate of 2-sulfanylethan-1-ol.

Genetic and metabolic engineering of microorganisms for the development of new flavor compounds from terpenic substrates.

Throughout human history, natural products have been the basis for the discovery and development of therapeutics, cosmetic and food compounds used in industry. Many compounds found in natural organisms are rather difficult to chemically synthesize and to extract in large amounts, and in this respect, genetic and metabolic engineering are playing an increasingly important role in the production of these compounds, such as new terpenes and terpenoids, which may potentially be used to create aromas in industry. Terpenes belong to the largest class of natural compounds, are produced by all living organisms and play a fundamental role in human nutrition, cosmetics and medicine. Recent advances in systems biology and synthetic biology are allowing us to perform metabolic engineering at the whole-cell level, thus enabling the optimal design of microorganisms for the efficient production of drugs, cosmetic and food additives. This review describes the recent advances made in the genetic and metabolic engineering of the terpenes pathway with a particular focus on systems biotechnology.

Natural products: Perspectives in the pharmacological treatment of gastrointestinal anisakiasis

Objective: To evaluate the activity of different natural products against L3 larvae of Anisakis type I. Methods: Information on investigations into the activity of natural products against the L3 larvae of Anisakis simplex was collected from scientific journals, books, theses and reports via a library and electronic search (using Pubmed, Scopus, Medline, Web of Science and ScienceDirect). The search terms included: natural products, medicinal plants, essential oils, terpenic derivatives, Anisakis, antinematodal activity. Results: In the literature reviewed numerous papers were found concerning the in vitro and in vivo activity against Anisakis type I of various natural products (plant extracts, essential oils and their major components). Analysis of the results showed that in vitro tests the Melaleuca alternifolia essential oil and the ar-turmerone isolated from Curcuma longa displayed the greatest activity. In vivo, the most active compound was perillaldehyde, the main component of Perilla frutescens essential oil. Conclusions: This study shows that some natural products exhibited promising antianisakis properties.

Phenolic profiles and antioxidant and anticarcinogenic activities of Greek herbal infusions; balancing delight and chemoprevention?

Total phenolic content, antioxidant activity and phenolic profiles of six herbal infusions - namely rosemary, Cretan dittany, St. John's Wort, sage, marjoram and thyme were assayed. Additionally, the infusion anticarcinogenic effect as to their ability to (a) scavenge free radicals, (b) inhibit cell growth, (c) decrease IL-8 levels and (d) regulate p65 subunit in epithelial colon cancer (HT29) and prostate (PC3) cancer cells was investigated. LC-DAD-MS and GC-MS analyses showed major qualitative and quantitative differences in phenolic profiles of the infusions. All herbal infusions exhibited antiradical activity which correlated strongly with their total phenolic content. Infusions exhibited the potential to inhibit cell growth and to reduce IL-8 levels in HT29 colon and PC3 prostate cancer cells. The regulation reported in p65 subunit in HT29 treated with St John's Wort and in PC3 treated with thyme might point to the NF-κB as the molecular target underlying the effect of these infusions.

Efectos antibacterianos de extractos de especies del género Ugni que crecen en Chile / Antibacterial effects of extracts of species of the genus Ugni that grow in Chile

Introducción: Ugni molinae Turcz., Ugni candollei (Barnéoud) O. Berg y Ugni selkirkii (Hook. & Arn.) Berg son arbustos que se distribuyen en el centro-sur de Chile, incluido el archipiélago de Juan Fernández. Dada su similitud morfológica parece que estas especies están estrechamente relacionadas, por lo tanto, podrían compartir compuestos químicos similares, como sustancias de tipo fenólico y del tipo terpénico, con reconocida actividad antimicrobiana, los que han sido identificados en Ugni molinae, la especie con mayor distribución en Chile. Objetivo: evaluar la composición química y la actividad antibacteriana de los extractos de las especies chilenas del género Ugni, y determinar si existen variaciones entre poblaciones que están creciendo separadas geográficamente y en diferentes hábitats, como en Chile continental y el archipiélago de Juan Fernández. Métodos: se determinó el contenido y la composición de compuestos químicos por métodos espectrofotométricos en extractos obtenidos con solventes de polaridad creciente. La actividad antibacteriana se evaluó a través de inclusión en agar tripticasa frente a Pseudomonas aeruginosa, Staphyloccoccus aureus y Enterobacter aerogenes. Resultados: Ugni candollei y Ugni selkirkii presentan la mayor concentración de fenoles, flavonoides y taninos totales. Con respecto a las poblaciones de Ugni molinae, la población del archipiélago de Juan Fernández presentó la mayor concentración de compuestos fenólicos y saponinas totales. Si bien existe una tendencia en la actividad antibacteriana en los extractos metanólicos, que se correlaciona con la composición química, no se consideran grandes diferencias entre estas. Conclusiones: dado que Ugni molinae comparte características morfológicas con otras especies del género que crecen en Chile, Ugni candollei y Ugni selkirkii, también comparte características químicas. Con respecto a las poblaciones de Ugni molinae, se observan diferencias en el contenido de compuestos fenólicos y saponinas totales, quizá por las características de hábitat donde estas especies y poblaciones están creciendo. La mezcla activa de los extractos metanólicos de especies del género Ugni y las poblaciones de Ugni molinae, llamada fitocomplejo, concentra las acciones antimicrobianas para las familias de componentes descritas, sin observarse grandes diferencias entre estas.

Introduction: Ugni molinae Turcz., Ugni candollei (Barneoud) O. Berg and Ugni selkirkii (Hook. & Arn.) Berg are shrubs growing in southern central Chile, including Juan Fernandez Islands. Given their morphological likeness, these species seem to be closely related. Therefore, they could share similar chemical compounds, such as phenolic and terpenic substances with well-known antimicrobial activity. These have been identified in Ugni molinae, the species with the broadest distribution in Chile. Objective: evaluate the chemical composition and antibacterial activity of extracts of Chilean species of the genus Ugni, and determine the possible differences between populations growing in geographically separate areas with dissimilar habitats, such as Continental Chile and Juan Fernandez Islands. Methods: the content and chemical composition of chemical compounds were determined by applying spectrophotometric methods to extracts obtained with increasing polarity solvents. Antibacterial activity against Pseudomonas aeruginosa, Staphylococcus aureus and Enterobacter aerogenes was evaluated by inclusion in trypticase agar. Results: Ugni candollei and Ugni selkirkii exhibit the greatest concentration of total tannins, phenols and flavonoids. With respect to Ugni molinae, the populations on Juan Fernandez Islands showed the greatest concentration of total saponins and phenolic compounds. Although there is a trend in the antibacterial activity of methanolic extracts that is consistent with their chemical composition, no great differences are found between them. Conclusions: Ugni molinae not only shares some morphological features with other species of the genus growing in Chile Ugni candollei and Ugni selkirkii. These species also share some chemical characteristics. With respect to Ugni molinae populations, differences are found in the content of total saponins and phenolic compounds which are maybe due to the specific habitat where these species and populations grow. The active mixture of methanolic extracts of species from the genus Ugni and Ugni molinae populations, known as phytocomplex, concentrates the antimicrobial activities of the families of components described above, and no great differences are found between them.