Molecular Characterization of Thymus capitellatus Extracts and Their Antioxidant, Neuroprotective and Anti-Proliferative Activities

Thymus capitellatus Hoffmanns & Link is an endemic species of the Iberian Peninsula listed as near-threatened, due to its restricted geographical distribution, occurring mainly in Portugal’s mainland. In this work, we detail for the first time T. capitellatus extracts’ phytochemical composition, as well as an evaluation of bioactivities to point out potential health benefits. Aqueous decoction (AD) and hydroethanolic (HE) extracts were obtained, both rich in flavonoids. However, quercetin-(?)-O-hexoside was identified as the main compound in T. capitellatus HE extract, while the phenolic acid rosmarinic acid was the main component of AD extracts. In addition, HE extract presents significant amounts of salvianolic acids and of the terpenoids oleanolic and ursolic acid. Both extracts showed antioxidant activity, evaluated by their capacity to scavenge ABTS and superoxide radicals, as well as an ability to prevent lipid peroxidation. AD extracts were also effective in scavenging hydroxyl and nitric oxide radicals. As potential functional foods, T. capitellatus extracts presented neuroprotective and anti-diabetic activity, in addition to time- and dose-dependent anti-proliferative activity against Caco-2 (colorectal adenocarcinoma) and HepG2 (hepatic carcinoma) cells. HE extract presented higher cytotoxicity than AD extract, and HepG2 cells were more resistant than Caco-2 cells. After 24 h exposure to HE extract, the IC$_{50}$ values were 330 μg/mL and 447 μg/mL for Caco-2 and HepG2 cells, respectively. T. capitellatus has potential as a functional food or as a source of bioactive molecules. These results also highlight the need to preserve species with as yet unknown molecular compositions and potential medicinal applications

Chemical composition and potential biological activity of melanoidins from instant soluble coffee and instant soluble barley: A comparative study

In this work a comparative study of the chemical composition and potential biological activity of high molecular weight (HMW) melanoidins isolated from instant soluble coffee (ISC) and instant soluble barley (ISB) was performed. ISB HMW melanoidins were almost exclusively composed by an ethanol soluble (EtSn) melanoidin fraction composed by glucose (76%w/w) partially susceptible to in vitro digestion, whereas ISC was composed mainly by arabinogalactans (∼41% w/w) and lower amounts of galactomannans (∼14%w/w) presenting a range of ethanol solubilities and resistant to in vitro digestion. Melanoidins from ISC presented a significantly higher content of condensed phenolic compounds (17/100 g) when compared to ISB (8/100 g) showing also a higher in vitro scavenging of ABTS•+ (329 mmol Trolox/100 g vs. 124 mmol Trolox/100 g) and NO radicals (inhibition percentage of 57 and 26%, respectively). Nevertheless, ISB EtSn melanoidins presented, on average a higher inhibitory effect on NO production from LPS-stimulated macrophages. ISB melanoidins, up to 1 mg/mL, did not induce toxicity in Caco-2, HepG2 and RAW 264.7 cell lines while at the highest concentration ISC slightly reduced cell viability. Thus, consumption of a diet rich in ISC and ISB melanoidins may reduce the oxidative stress, the inflammatory levels and increase the protective effects against chronic inflammatory diseases

Organic compounds isolated from Juniperus brevifolia bark

10º Encontro Nacional de Química Orgânica e 1º Simpósio Luso-Brasileiro de Química Orgânica, Lisboa, 4-6 Setembro de 2013In continuation of our study on the chemical characterization of endemic plants of the Azores archipelago, we have examined Juniperus brevifolia. Our interest in this plant was stimulated by the wide range of biological activities reported for other species of this genus and/or of their constituents. Previous work showed that leaves extracts, for instance dichloromethane extract and chloroform-soluble fraction of the acetone extract, showed high activity against HeLa and Hep-2 tumour cell lines. Chemical investigation of these extracts afforded more than thirty compounds, mainly abietane and pimarane derivatives, eight of them were new natural compounds and other exhibit interesting antitumor activity.These findings prompted us to study bark extracts.Thanks are due to Fundação para a Ciência e a Tecnologia (FCT, Portugal), the European Union, QREN, FEDER, COMPETE, for funding the Organic Chemistry Research Unit (QOPNA) (project PEst-C/QUI/UI0062/2011) and the Portuguese National NMR Network (RNRMN). M. Martins is grateful to CNPq, Conselho Nacional de Desenvolvimento Científico e Tecnológico – Brasil, for the scholarship granted

An implantable microsystem for electrical resistance and temperature measurements in cows with wireless capabilities suitable for reproductive management

In the cattle breeding industry, where artificial insemination techniques are employed, the successful detection of oestrus onset leads to considerable cost saving in herd management. One of the most reliable approaches is based on the determination of progesterone concentration in milk. However, these methods rely on the biosensor concept where a biological substrate is used in a chemical binding reaction to, directly or indirectly; produce some effect (electrical or light) that is used at the transducer level. These methods present several drawbacks concerning real-time measurements due to the complexity of the reactions involved and reagent/waste handling. Another approach is to combine measurements of electrical resistance of vaginal mucus and temperature to predict estrus. Using a low-power microsystem with wireless capabilities it is possible to take these measurements in-situ and more frequently. The proposed microsystem comprises a second-order delta-sigma modulator for analog to digital conversion and a class-E radio-frequency (RF) transmitter operating in the ISM-band of 433 MHz to transfer acquired data to a collar. Electrical resistance is measured by using a modified Wenner array and temperature by the on-chip temperature sensor. System (including battery and antenna) package is made of a tissue compatible material to allow implantation in the cow vulvar muscle

Phytochemical Composition and Antioxidant, Anti-Acetylcholinesterase, and Anti-α-Glucosidase Activity of Thymus carnosus Extracts: A Three-Year Study on the Impact of Annual Variation and Geographic Location

Thymus carnosus Boiss. is a near-threatened species, and, as for many species, its potential for medicinal purposes may be lost if measures towards plant protection are not taken. A way of preserving these species is to increase knowledge about their medicinal properties and economic potential. Thus, with the objective of studying the potentiality of introducing T. carnosus as a crop, the stability of the phytochemical profile of T. carnosus was studied during a period of three years by comparing the phytochemical profile of extracts obtained from plants harvested in two different edaphoclimatic locations, as well as by comparing the respective bioactivities, namely, antioxidant, antidiabetic, antiaging, and neuroprotective activities. It was reported, for the first time, the effect of annual variation and geographic location in the phytochemical composition of aqueous decoction and hydroethanolic extracts of T. carnosus. In addition, the presence of two salvianolic acid B/E isomers in T. carnosus extracts is here described for the first time. Despite the variations in phytochemical composition, according to harvesting location or year, T. carnosus extracts maintain high antioxidant activity, assessed by their capacity to scavenge ABTS•+, •OH , NO•, O2•− radicals, as well as to prevent β-carotene bleaching. All extracts presented significant potential to inhibit acetylcholinesterase (AChE), tyrosinase, and α-glucosidase, denoting neuroprotective, anti-aging, and anti-diabetic potential. In conclusion, the vegetative stage and location of harvest are key factors to obtain the maximum potential of this species, namely, a phytochemical profile with health benefit bioactivities

Genotoxicity assessment of metal-based nanocomposites applied in drug delivery

Nanocomposites as drug delivery systems (e.g., metal nanoparticles) are being exploited for several applications in the biomedical field, from therapeutics to diagnostics. Green nanocomposites stand for nanoparticles of biocompatible, biodegradable and non-toxic profiles. When using metal nanoparticles for drug delivery, the question of how hazardous these virus-sized particles can be is posed, due to their nanometer size range with enhanced reactivity compared to their respective bulk counterparts. These structures exhibit a high risk of being internalized by cells and interacting with the genetic material, with the possibility of inducing DNA damage. The Comet Assay, or Single-Cell Gel Electrophoresis (SCGE), stands out for its capacity to detect DNA strand breaks in eukaryotic cells. It has huge potential in the genotoxicity assessment of nanoparticles and respective cells interactions. In this review, the Comet assay is described, discussing several examples of its application in the genotoxicity evaluation of nanoparticles commonly administered in a set of routes (oral, skin, inhaled, ocular and parenteral administration). In the nanoparticles boom era, where guidelines for their evaluation are still very limited, it is urgent to ensure their safety, alongside their quality and efficacy. Comet assay or SCGE can be considered an essential tool and a reliable source to achieve a better nanotoxicology assessment of metal nanoparticles used in drug delivery.This work was funded by the Portuguese Science and Technology Foundation (FCT) from the Ministry of Science and Technology (MCTES), through the projects UIDB/04469/2020 (CEB strategic fund) and UIDB/04033/2020 (CITAB), co-funded by European Funds (PRODER/COMPETE) and FEDER, under the Partnership Agreement PT2020. The work was also supported by the National Science Centre within the MINIATURA 4 for single research activity (grant No: 2020/04/X/ST5/00789) and by the START 2021 Program of the Foundation for Polish Science (FNP) granted to Dr. Aleksandra Zielińskainfo:eu-repo/semantics/publishedVersio

Major anthocyanins in elderberry effectively trap methylglyoxal and reduce cytotoxicity of methylglyoxal in HepG2 cell line

The accumulation of advanced glycation end-products (AGEs) in the body is implicated in numerous diseases, being methylglyoxal (MGO) one of the main precursors. One of the strategies to reduce AGEs accumulation might be acting in an early stage of glycation by trapping MGO. Thus, this work aimed to evaluate, for the first time, the potential of elderberries polyphenols to trap MGO, access the formation of MGO adducts, and evaluate the cytoprotection effect in HepG2 and Caco-2 cells. The results demonstrated that monoglycosylated anthocyanins (cyanidin-3-glucoside and cyanidin-3-sambubioside) are very efficient in trapping MGO, forming mono- and di-adducts. Quercetin-3-glucoside and quercetin-3-rutinoside reacted slowly, while diglycosylated anthocyanins did not react. The trapping of MGO by elderberry monoglycosylated anthocyanins significantly decreased the MGO cytotoxicity in HepG2 cells (∼70 % of cell viability), while the effect in Caco-2 cells was lower (∼50 %). Thus, elderberry phenolics present antiglycation potential by trapping MGOpublishe

Thymus zygis subsp. zygis an endemic portuguese plant: phytochemical profiling, antioxidant, anti-proliferative and anti-inflammatory activities

Thymus zygis subsp. zygis is an endemic Portuguese plant belonging to the Thymus zygis species. Although T. zygis is commonly used as a condiment and as a medicinal herb, a detailed description of the polyphenol composition of hydroethanolic (HE) and aqueous decoction (AD) extracts is not available. In this work, we describe for the first time a detailed phenolic composition of Thymus zygis subsp. zygis HE and AD extracts, together with their antioxidant, anti-proliferative and anti-inflammatory activities. Unlike other Thymus species, T. zygis subsp. zygis extracts contain higher amounts of luteolin-(?)-O-hexoside. However, the major phenolic compound is rosmarinic acid, and high amounts of salvianolic acids K and I were also detected. T. zygis subsp. zygis extracts exhibited significant scavenging activity of ABTS+, hydroxyl (•OH), and nitric oxide (NO) radicals. Regarding the anti-proliferative/cytotoxic effect, tested against Caco-2 and HepG2 cells, the AD extract only slightly reduced cell viability at higher concentrations (IC50 > 600 µg/mL, 48 h exposure), denoting very low toxicity, while the HE extract showed a high anti-proliferative effect, especially at 48 h exposure (IC50 of 85.01 ± 15.10 μg/mL and 82.19 ± 2.46 μg/mL, for Caco-2 and HepG2, respectively). At non-cytotoxic concentrations, both extracts reduced the nitric oxide (NO) release by lipopolysaccharide (LPS)-stimulated RAW 264.7 cells (at 50 μg/mL, HE and AD extracts inhibited NO release in ~89% and 48%, respectively). In conclusion, the results highlight the non-toxic effect of aqueous extracts, both resembling the consumption of antioxidants in foodstuff or in functional food. Furthermore, the HE extract of T. zygis subsp. zygis is a source of promising molecules with antioxidant, anti-inflammatory and anticancer activities, highlighting its potential as a source of bioactive ingredients for nutraceutical and pharmaceutical industries.This work was supported by the INTERACT project–“Integrative Research in Environment, Agro-Chains and Technology”, no. NORTE-01-0145- FEDER-000017, in its line of research entitled ISAC, co-financed by the European Regional Development Fund (ERDF) through NORTE 2020 (North Regional Operational Program 2014/2020). By funds from the Portuguese Science and Technology Foundation, Ministry of Science and Education (FCT/MEC) through national funds, under the projects UIDB/04033/2020 (CITAB), UIDB/00616/2020 (CQ-VR) and UIDB/04469/2020 (CEB). FCT is also acknowledged for the grant to C.M.G. (SFRH/BD/145855/2019).info:eu-repo/semantics/publishedVersio

Labdanum resin from Cistus ladanifer L. as a source of compounds with anti-diabetic, neuroprotective and anti-proliferative activity

Labdanum resin or “gum” can be obtained from Cistus ladanifer L. by two different extraction methods: the Zamorean and the Andalusian processes. Although its main use is in the fragrance and perfumery sectors, ethnobotanical reports describe its use for medicinal purposes in managing hyperglycemia and mental illnesses. However, data concerning the bioactivities and pharmacological applications are scarce. In this work, it was found that the yield of labdanum resin extracted by the Andalusian process was 25-fold higher than the Zamorean one. Both resins were purified as absolutes, and the Andalusian absolute was purified into diterpenoid and flavonoid fractions. GC-EI-MS analysis confirmed the presence of phenylpropanoids, labdane-type diterpenoids, and methylated flavonoids, which are already described in the literature, but revealed other compounds, and showed that the different extracts presented distinct chemical profile. The potential antidiabetic activity, by inhibition of α-amylase and α-glucosidase, and the potential neuroprotective activity, by inhibition of acetylcholinesterase, were investigated. Diterpenoid fraction produced the higher α-amylase inhibitory effect (~30% and ~40% at 0.5 and 1 mg/mL, respectively). Zamorean absolute showed the highest α-glucosidase inhibitory effect (~14% and ~24%, at 0.5 and 1 mg/mL, respectively). Andalusian absolute showed the highest acetylcholinesterase inhibitory effect (~70% and ~75%, at 0.5 and 1 mg/mL, respectively). Using Caco-2 and HepG2 cell lines, Andalusian absolute and its purified fractions showed moderate cytotoxic/anti-proliferative activity at 24 h exposure (IC50 = 45–70 µg/mL, for Caco-2; IC50 = 60–80 µg/mL, for HepG2), whereas Zamorean absolute did not produce cytotoxicity (IC50 ≥ 200.00 µg/mL). Here we show, for the first time, that labdanum resin obtained by the Andalusian process, and its fractions, are composed of phytochemicals with anti-diabetic, neuroprotective and anti-proliferative potential, which are worth investigating for the pharmaceutical industry. However, toxic side-effects must also be addressed when using these products by ingestion, as done traditionally.N/

Biosurfactants: properties and applications in drug delivery, biotechnology and ecotoxicology

Surfactants are amphiphilic compounds having hydrophilic and hydrophobic moieties in their structure. They can be of synthetic or of microbial origin, obtained respectively from chemical synthesis or from microorganisms activity. A new generation of ecofriendly surfactant molecules or biobased surfactants is increasingly growing, attributed to their versatility of applications. Surfactants can be used as drug delivery systems for a range of molecules given their capacity to create micelles which can promote the encapsulation of bioactives of pharmaceutical interest; besides, these assemblies can also show antimicrobial properties. The advantages of biosurfactants include their high biodegradability profile, low risk of toxicity, production from renewable sources, functionality under extreme pH and temperature conditions, and long-term physicochemical stability. The application potential of these types of polymers is related to their properties enabling them to be processed by emulsification, separation, solubilization, surface (interfacial) tension, and adsorption for the production of a range of drug delivery systems. Biosurfactants have been employed as a drug delivery system to improve the bioavailability of a good number of drugs that exhibit low aqueous solubility. The great potential of these molecules is related to their auto assembly and emulsification capacity. Biosurfactants produced from bacteria are of particular interest due to their antibacterial, antifungal, and antiviral properties with therapeutic and biomedical potential. In this review, we discuss recent advances and perspectives of biosurfactants with antimicrobial properties and how they can be used as structures to develop semisolid hydrogels for drug delivery, in environmental bioremediation, in biotechnology for the reduction of production costs and also their ecotoxicological impact as pesticide alternative.This work was supported by the Portuguese Science and Technology Foundation (FCT/MCT) and from European Funds (PRODER/COMPETE) through the grants M-ERA-NET/0004/2015 (PAIRED), UIDB/04469/2020 (strategic fund), and UIDB/04033/2020 (CITAB), cofinanced by FEDER, under the Partnership Agreement PT2020. T. Pashirova acknowledges the financial support received from the government assignment for FRC Kazan Scientific Center of Russian Academy of Sciences.info:eu-repo/semantics/publishedVersio