P80 natural essence spray and lozenges provide respiratory protection against Influenza A, B, and SARS-CoV-2.

Seasonally circulating viruses, such as Influenza, as well as newly emerging viruses and variants thereof, and waning immunity urge the need for safe, easy-to-use and inexpensive drugs to protect from these challenges. To prevent transmission of these viruses and subsequent excessive inflammatory reactions on mucous membranes, we tested the efficacy of the natural essence P80 as spray and in form of lozenges against respiratory infections caused by SARS-CoV-2 variants of concern (VoCs), influenza A (H3N2) and influenza B (Victoria). P80 natural essence, a Dimocarpus longan extract, shielded highly differentiated human airway epithelia from SARS-CoV-2 wildtype and Omicron variant as well as Influenza A and B infection and dampened inflammation by down-modulating pro-inflammatory cytokine and anaphylatoxin secretion. A single application of P80 natural essence spray maintained tissue integrity long-term. This also significantly reduced the release of infectious viral particles and the secretion of IP10, MCP1, RANTES and C3a, all of which mediate the migration of immune cells to the sites of infection. Even P80 lozenges dissolved in distilled water or non-neutralizing saliva efficiently prevented SARS-CoV-2 and Influenza-induced tissue destruction. Consequently, our in vitro data suggest that P80 natural essence can act as antiviral prophylactic, both in form of nasal or oral spray and in form of lozenges, independent of circulating respiratory challenges.

Phytochemical Profiling, Antioxidant and Tyrosinase Regulatory Activities of Extracts from Herb, Leaf and In Vitro Culture of Achillea millefolium (Yarrow).

Achillea millefolium L. is one of the most known medicinal plants with a broad spectrum of applications in the treatment of inflammation, pain, microbial infections and gastrointestinal disorders. In recent years, the extracts from A. millefolium have also been applied in cosmetics with cleansing, moisturizing, shooting, conditioning and skin-lightening properties. The growing demand for naturally derived active substances, worsening environmental pollution and excessive use of natural resources are causing increased interest in the development of alternative methods for the production of plant-based ingredients. In vitro plant cultures are an eco-friendly tool for continuous production of desired plant metabolites, with increasing applicability in cosmetics and dietary supplements. The purpose of the study was to compare phytochemical composition and antioxidant and tyrosinase inhibitory properties of aqueous and hydroethanolic extracts from A. millefolium obtained from field conditions (AmL and AmH extracts) and in vitro cultures (AmIV extracts). In vitro microshoot cultures of A. millefolium were obtained directly from seeds and harvested following 3 weeks of culture. Extracts prepared in water, 50% ethanol and 96% ethanol were compared for the total polyphenolic content, phytochemical content using the ultra-high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-hr-qTOF/MS), antioxidant activity by DPPH scavenging assay and the influence on the activity of mushroom and murine tyrosinases. The phytochemical content of AmIV extracts was significantly different from AmL and AmH extracts. Most of the polyphenolic compounds identified in AmL and AmH extracts were present in AmIV extracts only in trace amounts and the major constituents presented in AmIV extracts were fatty acids. The total content of polyphenols in AmIV exceeded 0.25 mg GAE/g of dried extract, whereas AmL and AmH extracts contained from 0.46 ± 0.01 to 2.63 ± 0.11 mg GAE/g of dried extract, depending on the solvent used. The low content of polyphenols was most likely responsible for the low antioxidant activity of AmIV extracts (IC50 values in DPPH scavenging assay >400 µg/mL) and the lack of tyrosinase inhibitory properties. AmIV extracts increased the activity of mushroom tyrosinase and tyrosinase present in B16F10 murine melanoma cells, whereas AmL and AmH extracts showed significant inhibitory potential. The presented data indicated that microshoot cultures of A. millefolium require further experimental research before they can be implemented as a valuable raw material for the cosmetics industry.

Ambient mass spectrometry and near-infrared spectroscopy - a direct comparison of methods for the quantification of sucralose in e-liquids.

E-liquids have become increasingly popular in society in recent years. A wide variety of flavors and nicotine strengths make it possible for every user to get a product according to their wishes. Many of these e-liquids are marketed with countless different flavors, which are often characterized by a strong and sweet smell. Sweeteners, such as sucralose, are therefore commonly added as sugar substitutes. However, recent studies have shown the potential formation of highly toxic chlorinated compounds. This can be explained by the high temperatures (above 120 °C) within the heating coils and the used basic composition of these liquids. Nevertheless, the legal situation is composed of proposals without clear restrictions, only recommendations for tobacco products. For this reason, a high level of interest lies within the establishment of fast, reliable and cost-effective methods for the detection of sucralose in e-liquids. In this study, a number of 100 commercially available e-liquids was screened for sucralose in order to identify the suitability of ambient mass spectrometry and near-infrared spectroscopy for this application. A highly sensitive high-performance liquid chromatography coupled to a tandem mass spectrometer method was used as reference method. Furthermore, the advantages and limitations of the two mentioned methods are highlighted in order to provide a reliable quantification of sucralose. The results clearly revile the necessity for product quality due to the absence of declaration on many of the used products. Further on, it could be shown, that both methods are suitable for the quantification of sucralose in e-liquids, with beneficial economic and ecological aspects, over classical analytical tools including high-performance liquid chromatography. Clear correlations between the reference and novel developed methods are displayed. In summary, these methods enable an important contribution to ensure consumer protection and elimination of confuse package labelling.

GlyPerA™ effectively shields airway epithelia from SARS-CoV-2 infection and inflammatory events.

New SARS-CoV-2 variants of concern (VOCs) and waning immunity illustrate that quick and easy-to-use agents are needed to prevent infection. To protect from viral transmission and subsequent inflammatory reactions, we applied GlyperA™, a novel antimicrobial formulation that can be used as mouth gargling solution or as nasal spray, to highly differentiated human airway epithelia prior infection with Omicron VOCs BA.1 and BA.2. This formulation fully protected polarized human epithelium cultured in air-liquid interphase (ALI) from SARS-CoV-2-mediated tissue destruction and infection upon single application up to two days post infection. Moreover, inflammatory reactions induced by the Omicron VOCs were significantly lowered in tissue equivalents either pre-treated with the GlyperA™ solution, or even when added simultaneously. Thus, the GlyperA™ formulation significantly shielded epithelial integrity, successfully blocked infection with Omicron and release of viral particles, and decreased intracellular complement C3 activation within human airway epithelial cell cultures. Crucially, our in vitro data imply that GlyperA™ may be a simple tool to prevent from SARS-CoV-2 infection independent on the circulating variant via both, mouth and nose.

Rapid quality control of black truffles using Direct Analysis in Real Time Mass Spectrometry and Hydrophilic Interaction Liquid Chromatography Mass Spectrometry.

This work is the first to describe the application of Direct Analysis in Real Time Mass Spectrometry for rapid and simple quality control of black truffles. Multivariate analyses (PCA and LDA) of mass spectra provided highly efficient models for the discrimination of four truffle species, namely Tuber melanosporumVittad., Tuber aestivumVittad., Tuber uncinatumChatin and Tuber indicum Cooke etMassee. Full-cross validations showed prediction accuracies up to 99%. Hydrophilic Interaction Liquid Chromatography Mass Spectrometry was used as a reference method and the results of both methods were compared to each other. Multivariate models of the chromatograms also showed excellent results with prediction accuracies of 100%. In a direct comparison of methods, Direct Analysis in Real Time Mass Spectrometry showed minimally lower prediction accuracies, but was significantly faster, more robust and easier to use, making it an excellent choice for rapid quality control of black truffles.

Gastroprotective effects of Caragana ambigua stocks on ethanol-induced gastric ulcer in rats supported by LC-MS/MS characterization of formononetin and biochanin A.

BACKGROUND: Caragana ambigua has been the part of the dietary routines of the regional people in south-west Pakistan and has traditionally been used for the treatment of diabetes there. There is an increased production of reactive oxygen species in diabetics, leading to gastrointestinal disorders. Natural antioxidants exhibit gastroprotective effects owing to their free-radical scavenging action. C. ambigua possesses appreciable phenolic and flavonoid content; thus, it has the potential to protect against gastrointestinal disorders (e.g. gastric ulcer). RESULTS: This study reports the anti-ulcer potential of C. ambigua. Four different fractions (chloroform, ethyl acetate, butanol, and aqueous) of plant were compared against omeprazole. Ulcer index, ulcer inhibition percentage, gastric pH and volume, total acidity, gastric protein, gastric wall mucus, and histopathology of gastric walls of rats were assessed. All fractions exhibited a reduction in ulcer index and promotion of percentage of ulcer inhibition compared with the ulcer control group. Furthermore, the fractions revealed a significant (P < 0.001) diminution in gastric volume and total acidity with an increase in pH. Among the fractions investigated, the chloroform fraction unveiled the most promising anti-ulcer activity, which is comparable to omeprazole. Liquid chromatography-tandem mass spectrometry screening of fractions revealed the presence of formononetin and biochanin A (isoflavones reported to have anti-ulcer properties) in the chloroform fraction. CONCLUSION: This study establishes that C. ambigua possesses significant potential in reducing gastric ulcer progression. Formononetin and biochanin A are chiefly responsible for the stated bioactivity due to the fact that these compounds were solely present in the chloroform fraction. © 2022 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Anti-Inflammatory Extract from Soil Algae Chromochloris zofingiensis Targeting TNFR/NF-κB Signaling at Different Levels.

Inflammatory skin diseases, including atopic dermatitis (AD) and psoriasis, are increasing in populations worldwide. The treatment of patients with AD and other forms of skin inflammation is mainly based on the use of topical corticosteroids or calcineurin inhibitors, which can cause significant side effects with long-term use. Therefore, there is a great need for the development of more effective and less toxic anti-inflammatory agents suitable for the treatment of chronic skin lesions. Here, we screened a number of strains from the ASIB 505 terrestrial algae collection and identified a green algae Chromochloris zofingiensis with pronounced anti-inflammatory properties. We found that a crude nonpolar extract of C. zofingiensis (ID name NAE_2022C), grown upon nitrogen deprivation, acts as a bioactive substance by inhibiting TNFR/NF-κB responses in human skin keratinocyte HaCaT cells. We also found that NAE_2022C suppressed the secretion of pro-inflammatory cytokine tumor necrosis factor α (TNFα) and several Th1- and Th2-related chemokines in a reconstituted human epidermis. The TNFR/NF-κB pathway analysis showed multiple inhibitory effects at different levels and disclosed a direct targeting of IKKß by the extract. Bioassay-guided fractionation followed by high-resolution mass spectrometry detected diacylglyceryl-trimethylhomoserine (DGTS), Lyso-DGTS (LDGTS), 5-phenylvaleric acid, theophylline and oleamide as leading metabolites in the active fraction of NAE_2022C. Further analysis identified betaine lipid DGTS (32:0) as one of the active compounds responsible for the NAE_2022C-mediated NF-κB suppression. Overall, this study presents an approach for the isolation, screening, and identification of anti-inflammatory secondary metabolites produced by soil algae.

A validated method for the rapid quantification of melatonin in over-the-counter hypnotics by the atmospheric pressure solid analysis probe (ASAP).

Melatonin is a hormone that regulates the biological day and night cycle. It is mainly produced by the pineal gland during the night. People suffering from insomnia use it as a soporific drug. The aim of this study was to develop a method for the rapid quantification of melatonin in hypnotics. For that purpose, atmospheric pressure solid analysis probe-assisted mass spectrometry was applied, where no chromatographic separation is needed. Thereby, one single analysis takes less than 1 min. Reference measurements were performed with ultra-high-performance liquid chromatography coupled with a quadrupole-time-of-flight mass spectrometer. Both methods were validated and real sample extracts were tested. The coefficients of determination were above 0.97 for both methods. The limits of detection and quantification were below 1 mg kg-1. Both methods gave comparable results. Moreover, the content of melatonin differed from the specified value in many samples. The highest and lowest observed deviations were 78% and 1%, respectively.

Fast and semiquantitative screening for sildenafil in herbal over-the-counter formulations with atmospheric pressure solid analysis probe (ASAP) to prevent medicinal adulteration.

Herbal medicines are commonly used in many countries all around the world. In Western countries they are now gaining more and more popularity, whereas in countries like China and India they have been entrenched for millenniums. Some of these perceived herbal medicines claim to help when suffering from erectile dysfunction. Nevertheless, many of these products are adulterated with PDE5 inhibitors like sildenafil or α-blockers. Patients who suffer from high blood pressure sometimes resort to herbal products, as they are not allowed to take sildenafil because of negative drug-drug interactions with nitrates (often utilized as treatment for coronary diseases). Products which are then adulterated with PDE5 inhibitors, can seriously harm patients. Therefore, this study reports the instant screening of alleged herbal products by employing atmospheric pressure solids analysis probe and high-resolution mass spectrometry to determine adulterants. Three out of 12 investigated products contained sildenafil in ranges from 0.5% to 18%. Multivariate analysis of ambient mass spectrometry measurements revealed encouraging outcomes for distinguishing non-sildenafil and sildenafil adulterated samples. Atmospheric pressure solids analysis probe is therefore a promising method for the rapid determination of sildenafil in herbal products with possible downstream semiquantitative analysis.

Phenylpropanoid Glycoside and Phenolic Acid Profiles and Biological Activities of Biomass Extracts from Different Types of Verbena officinalis Microshoot Cultures and Soil-Grown Plant.

Different types of microshoot cultures (agar, stationary liquid, agitated, and bioreactors) of Verbena officinalis were optimized for biomass growth and the production of phenylpropanoid glycosides and phenolic acids. Using ultra-high performance liquid chromatography with high-resolution time-of-flight mass spectrometry, the presence of verbascoside, isoverbascoside, leucoseptoside A/isomers, and cistanoside D/isomer was confirmed in the methanolic extracts obtained from all types of in vitro cultures. The compound's content was determined by ultra-high-performance liquid chromatography. The main metabolites in biomass extracts were verbascoside and isoverbascoside (maximum 4881.61 and 451.80 mg/100 g dry weight (DW)). In the soil-grown plant extract, verbascoside was also dominated (1728.97 mg/100 g DW). The content of phenolic acids in the analyzed extracts was below 24 mg/100 g DW. The highest radical scavenging activity was found in the biomass extract from agitated cultures, the most effective reducing power in agar culture extract, and the highest chelating activity in extract from bioreactor cultures. The extracts showed significantly stronger bacteriostatic and bactericidal activity against Gram-positive bacteria (minimum inhibitory concentration (MIC) of 0.3-2.2 mg/mL and minimum bactericidal concentration (MBC) of 0.6-9 mg/mL) than against Gram-negative bacteria (MIC 0.6-9 mg/mL, MBC of 0.6-18 mg/mL). The biomass extract from liquid stationary culture showed the strongest antibacterial activity, while the extract from soil-grown herb had the lowest.

Rapid discrimination of Curcuma longa and Curcuma xanthorrhiza using Direct Analysis in Real Time Mass Spectrometry and Near Infrared Spectroscopy.

This study describes a newly developed method for the fast and straightforward differentiation of two turmeric species using Direct Analysis in Real Time mass spectrometry and miniaturized Near Infrared spectroscopy. Multivariate analyses (PCA and LDA) were performed on the mass spectrometric data, thus creating a powerful model for the discrimination of Curcumalonga and Curcumaxanthorrhiza. Cross-validation of the model revealed correctness-scores of 100% with 20-fold as well as leave-one-out validation techniques. To further estimate the models prediction power, seven retail samples of turmeric powder were analyzed and assorted to a species. Looking for a fast, non-invasive, cost-efficient and laboratory independent method, miniaturized NIR spectrometers offer an alternative for quality control of turmeric species. However, different technologies implemented to compensate for their small size, lead to different applicability of these spectrometers. Therefore, we investigated the three handheld spectrometers microPHAZIR, MicroNIR 2200 and MicroNIR 1700ES for their application in spice analysis in hyphenation to PCA, LDA and ANN methods used for the discriminant analysis. While microPHAZIR proved to be the most valuable device for differentiating C.longa and C.xanthorrhiza, MicroNIR 1700ES offered the worst results. These findings are interpreted on the basis of a quantum chemical simulation of the NIR spectrum of curcumin as the representative constituent. It was found that the information accessible to MicroNIR 1700ES that is relevant to the analyzed constituents is located in the spectral region prone to interferences with the matrix, likely limiting the performance of this spectrometer in this analytical scenario.

Rapid differentiation and quality control of tobacco products using Direct Analysis in Real Time Mass Spectrometry and Liquid Chromatography Mass Spectrometry.

This work is the first to describe the use of Direct Analysis in Real Time mass spectrometry (DART-MS) for the rapid and cost-effective quality control of cigars. Multivariate analyses (PCA and LDA) on mass spectra led to highly efficient models for the discrimination of tobacco products. In addition to the geographic origin of the cigars, different pretreatments of the tobacco leaves could also be determined. Cross-validations of the models yielded prediction accuracies up to 92.18%. All results were compared using LC-MS as reference method. Multivariate analyses of chromatograms also yielded powerful models with correctness scores up to 88.1%. However, the models generated with LC-MS were neither able to determine the geographic origin of the tobacco leaves nor different pretreatments. In addition to qualitative analysis, the nicotine content of tobacco leaves was also determined via both methods. While precise quantification was not possible with DART-MS, a correlation coefficient of >0.96 was achieved in direct comparison with LC-MS, which allows semi-quantitative statements about the nicotine content without any problems.

Quantification and cytotoxicity of degradation products (chloropropanols) in sucralose containing e-liquids with propylene glycol and glycerol as base.

Electronic cigarettes (e-cigarettes) have gained increasing popularity in recent years, mostly because they are supposed to be less harmful than regular cigarettes. Therefore, it is highly imperative to investigate possible noxious effects to protect the consumers. E-liquids consist of propylene glycol, glycerol, aroma compounds and sweeteners. One of these sweeteners is a chlorinated version of sucrose, namely sucralose. The aim of this work was to investigate degradation products of sucralose in the presence of propylene glycol and glycerol at different temperatures of commercially available e-cigarettes. Chemical analysis and biological tests were simultaneously performed on e-liquid aerosol condensates. The results of the chemical analysis, which was executed by employing GC-MS/GC-FID, demonstrated high amounts of various chloropropanols. The most abundant one is extremely toxic, namely 3-chloropropane-1,2-diol, which can be detected at concentrations ranging up to 10,000 mg/kg. Furthermore, a cytotoxicity investigation of the condensates was performed on HUVEC/Tert2 cells in which metabolic activity was determined by means of resazurin assay. The cellular metabolic activity significantly decreased by treatment with e-liquid aerosol condensate. Due to the results of this study, we advise against the use of sucralose as sweetener in e-liquids.

Near-Infrared Spectroscopy as a Rapid Screening Method for the Determination of Total Anthocyanin Content in Sambucus Fructus.

Elderberry (Sambucus nigra L., fructus) is a very potent herbal drug, deriving from traditional European medicine (TEM). Ripe elderberries are rich in anthocyanins, flavonols, flavonol esters, flavonol glycosides, lectins, essential oils, unsaturated fatty acids and vitamins. Nevertheless, unripe elderflower fruits contain a certain amount of sambunigrin, a toxic cyanogenic glycoside, whose concentration decreases in the ripening process. Therefore, quality assurance must be carried out. The standard method described in literature is the photometric determination (pH-differential method) of the total anthocyanin content (TAC) that is the highest when the berries are ripe. The drawback of the pH-differential method is the extensive sample preparation and the low accuracy of the method. Therefore, the goal of this publication was to develop a fast non invasive near-infrared (NIR) method for the determination of TAC in whole berries. TAC of elderberries was measured using pH-differentiation method where TAC values of 632.87 mg/kg to 4342.01 mg/kg were measured. Additionally, cyanidin-3-O-glucoside, cyanidin-3-O-sambubioside and cyanidin-3-O-sambubioside-5-O-glucoside which represent more than 98% of TAC in elderberry were quantified using ultra high performance liquid chromatography-multiple wavelength detection-ultra high resolution-quadrupole-time of flight-mass spectrometry (UHPLC-MWD-UHR-Q-TOF-MS) and their sum parameter was determined, ranging between 499.43 mg/kg and 8199.07 mg/kg. Using those two methods as reference, whole elderberries were investigated by NIR spectroscopy with the Büchi NIRFlex N-500 benchtop spectrometer. According to the constructed partial least squares regression (PLSR) models the performance was as follows: a relative standard deviation (RSDPLSR) of 13.5% and root mean square error of calibration (RMSECV/RMSEC) of 1.31 for pH-differentiation reference and a RSDPLSR of 12.9% and RMSECV/RMSEC of 1.28 for the HPLC reference method. In this study, we confirm that it is possible to perform a NIR screening for TAC in whole elderberries. Using quantum chemical calculations, we obtained detailed NIR band assignments of the analyzed compounds and interpreted the wavenumber regions established in PLSR models as meaningful for anthocyanin content. The NIR measurement turned out to be a fast and cost-efficient alternative for the determination of TAC compared to pH-differential method and UHPLC-MWD-UHR-Q-TOF-MS. Due to the benefit of no sample preparation and extraction the technology can be considered as sustainable green technology. With the above mentioned inversely proportional ratio of TAC to total amount of toxic cyanogenic glycosides, NIR proves to be a reliable screening method for the ideal harvest time with maximal content of TAC and lowest content of cyanogenic glycosides in elderberry.

Complementary α-arrestin-ubiquitin ligase complexes control nutrient transporter endocytosis in response to amino acids.

How cells adjust nutrient transport across their membranes is incompletely understood. Previously, we have shown that S. cerevisiae broadly re-configures the nutrient transporters at the plasma membrane in response to amino acid availability, through endocytosis of sugar- and amino acid transporters (AATs) (Müller et al., 2015). A genome-wide screen now revealed that the selective endocytosis of four AATs during starvation required the α-arrestin family protein Art2/Ecm21, an adaptor for the ubiquitin ligase Rsp5, and its induction through the general amino acid control pathway. Art2 uses a basic patch to recognize C-terminal acidic sorting motifs in AATs and thereby instructs Rsp5 to ubiquitinate proximal lysine residues. When amino acids are in excess, Rsp5 instead uses TORC1-activated Art1 to detect N-terminal acidic sorting motifs within the same AATs, which initiates exclusive substrate-induced endocytosis. Thus, amino acid excess or starvation activate complementary α-arrestin-Rsp5-complexes to control selective endocytosis and adapt nutrient acquisition.

Determination of phototoxic furanocoumarins in natural cosmetics using SPE with LC-MS.

Furanocoumarins are a class of naturally occurring phototoxic substances widely spread throughout the Umbelliferae and Rutacea families of plants. Plant extract of these families (e.g. citrus extracts) are commonly used as ingredients in fragrances and cosmetic products. Due to their phototoxicity these substances are regulated within the European Union according to the European Regulation (EC) No. 1223/2009 on cosmetic products. The proposed analytical workflow aims to swiftly screen out and determine the amount of phototoxic furanocoumarins in natural cosmetics employing solid-phase extraction (SPE). Cosmetic samples were first extracted in methanol and subsequently diluted in 35% (v/v) methanol before being applied to solid-phase extraction. The extraction procedure allows extraction of the eight target compounds including the six marker-substances proposed by the International Fragrance Association (IFRA) from complex cosmetic samples with satisfactory recovery levels ranging from 84 to 116% with the exception of bergamottin (68-89%). HPLC-UV and ESI-MS were used for the analysis of the target compounds. Furanocoumarin quantification was performed by standard addition. The workflow was successfully validated according to the ISO 12787 international standard ("Cosmetics - Analytical methods- Validation criteria for analytical results using chromatographic techniques"). Results of the validation on standard solutions highlight a LOQ of 0.01 mg kg-1, acceptable linearity up to 1 mg/kg as well as good accuracy (Bias <15%) and precision (RSD <15%). Total furanocoumarin levels of the examined natural cosmetic products were in general in agreement with the European Regulation. However, it was shown that total levels of furanocoumarins might be significantly higher than assumed using the six proposed marker substances only.

Comparison survey of EVOO polyphenols and exploration of healthy aging-promoting properties of oleocanthal and oleacein.

Olive oil is widely accepted as a superior edible oil. Great attention has been given lately to olive oil polyphenols which are linked to significant health beneficial effects. Towards a survey of Greek olive oil focusing on polyphenols, representative extra virgin olive oils (EVOOs) from the main producing areas of the country and the same harvesting period have been collected and analyzed. Significant differences and interesting correlations have been identified connecting certain polyphenols namely hydroxytyrosol, tyrosol, oleacein and oleocanthal with specific parameters e.g. geographical origin, production procedure and cultivation practice. Selected EVOOs polyphenol extracts, with different oleacein and oleocanthal levels, as well as isolated oleacein and oleocanthal were bio-evaluated in mammalian cells and as a dietary supplement in the Drosophila in vivo model. We found that oleocanthal and oleacein activated healthy aging-promoting cytoprotective pathways and suppressed oxidative stress in both mammalian cells and in flies.

Identification and quantification and antioxidant activity of flavonoids in different strains of silk cocoon, Bombyx mori.

Silk cocoon is produced from silkworm (Bombyx mori) to protect itself from outer environment. Various strains of cocoon contain different forms and amounts of flavonoids, which may affect on their antioxidant activity. Moreover, the extraction method would influence the amount of flavonoids extracted. Therefore, the objectives of this study were to identify and quantify the flavonoids in 3 strains of bivoltine Bombyx mori silk cocoon (Chul 1/1; white cocoon, Chul 3/2; greenish cocoon, and Chul 4/2; yellow cocoon) extracted by 6 different solvents including acetone, ethyl acetate, dimethyl sulfoxide (DMSO), ethanol, methanol, and purified water. The flavonoids extracted were identified and quantified by liquid chromatography-mass spectrometry (LC-MS). The antioxidant activity of flavonoids extracted was also investigated by visible spectroscopy at 517 nm. The results showed that Chul 3/2 silk cocoon contained the highest amount of flavonoids. Purified water seemed to be the best solvent that preserved most antioxidant activity of the flavonoids extracted. Flavonoids in Chul 1/1 and Chul 4/2 silk cocoon were rarely found, however they contained some antioxidant activities. The data from this study can provide basic information for flavonoid extraction from silk cocoon which can also apply for other flavonoid-containing natural biomaterials.

Vapours of US and EU Market Leader Electronic Cigarette Brands and Liquids Are Cytotoxic for Human Vascular Endothelial Cells.

The present study was conducted to provide toxicological data on e-cigarette vapours of different e-cigarette brands and liquids from systems viewed as leaders in the e-cigarette market and to compare e-cigarette vapour toxicity to the toxicity of conventional strong high-nicotine cigarette smoke. Using an adapted version of a previously constructed cigarette smoke constituent sampling device, we collected the hydrophilic fraction of e-cigarette vapour and exposed human umbilical vein endothelial cells (HUVECs) to the mixture of compounds present in the vapour of 4 different single-use e-cigarettes, 6 different liquid vapours produced by the same refillable e-cigarette, and one e-cigarette with an exchangeable liquid cartridge. After incubation of cells with various concentrations and for various periods of time we analysed cell death induction, proliferation rates, the occurrence of intra-cellular reactive oxygen species, cell morphology, and we also measured e-cigarette heating coil temperatures. Overall, conventional cigarette smoke extract showed the most severe impact on endothelial cells. However, some e-cigarette vapour extracts showed high cytotoxicity, inhibition of cell proliferation, and alterations in cell morphology, which were comparable to conventional high-nicotine cigarettes. The vapours generated from different liquids using the same e-cigarette show substantial differences, pointing to the liquids as an important source for toxicity. E-cigarette vapour-mediated induction of oxidative stress was significant in one out of the 11 analysed vapours. There is a high variability in the acute cytotoxicity of e-cigarette vapours depending on the liquid and on the e-cigarettes used. Some products showed toxic effects close to a conventional high-nicotine cigarette. Liquid nicotine, menthol content, and the formation of acute intracellular reactive oxygen species do not seem to be the central elements in e-cigarette vapour toxicity.

Poly(N-vinylimidazole/ethylene glycol dimethacrylate) for the purification and isolation of phenolic acids.

In this study we report the novel polymeric resin poly(N-vinyl imidazole/ethylene glycol dimethacrylate) for the purification and isolation of phenolic acids. The monomer to crosslinker ratio and the porogen composition were optimized for isolating phenolic acids diluted in acetonitrile at normal phase chromatography conditions, first. Acetonitrile serves as polar, aprotic solvent, dissolving phenolic acids but not interrupting interactions with the stationary phase due to the approved Hansen solubility parameters. The optimized resin demonstrated high loading capacities and adsorption abilities particularly for phenolic acids in both, acetonitrile and aqueous solutions. The adsorption behavior of aqueous standards can be attributed to ion exchange effects due to electrostatic interactions between protonated imidazole residues and deprotonated phenolic acids. Furthermore, adsorption experiments and subsequent curve fittings provide information of maximum loading capacities of single standards according to the Langmuir adsorption model. Recovery studies of the optimized polymer in the normal-phase and ion-exchange mode illustrate the powerful isolation properties for phenolic acids and are comparable or even better than typical, commercially available solid phase extraction materials. In order to prove the applicability, a highly complex extract of rosemary leaves was purified by poly(N-vinyl imidazole/ethylene glycol dimethacrylate) and the isolated compounds were identified using UHPLC-qTOF-MS.