Phenolic Composition and Bioactivities of Invasive Ailanthus altissima (Mill.) Swingle Leaf Extracts Obtained by Two-Step Sequential Extraction.

Ailanthus altissima, a highly invasive species, contains valuable compounds in different plant parts, indicating great practical potential. This paper proposes the use of non-polar (n-hexane) and polar (ethanol) solvents for the extraction of antioxidant compounds from A. altissima (family Simaroubaceae) leaves in a sequential two-step process. Fresh and dried leaves were examined for their microstructure by scanning electron microscopy, and for color changes in the CIELAB color space co-ordinates. An investigation of the harvesting season, processing (freezing and drying), and solvent indicates ethanol can be used for the highly efficient extraction of phenolics, flavonoids, tannins, and carotenoids. Statistically significant differences were found between the autumn and summer samples for phenolic content, and between dried and frozen samples for tannin content. The HPLC phenolic profile indicates more phenolics (nine polyphenols) in dried leaves harvested in both seasons compared to those in frozen ones (five to six polyphenols). Frozen leaves showed a higher antioxidant activity in a ferric-reducing antioxidant power assay than that of the dried samples, which exhibited a higher antioxidant activity using the 1, 1-diphenyl-2-picryl-hydrazyl assay, but it was not statistically significant. The phenolic, flavonoid, and carotenoid contents significantly influenced the antioxidant activities. Among the ethanolic extracts, those from dried leaves showed better antibacterial activity, in particular, on Staphylococcus aureus and Enterococcus faecalis. The high bioactive content and activity of A. altissima leaves make them suitable natural raw materials for various applications.

In Vitro Screening of Ecotoxic and Cytotoxic Activities of Ailanthus altissima Leaf Extract against Target and Non-Target Plant and Animal Cells.

Ailanthus altissima, an invasive plant species, exhibits pharmacological properties, but also some allergic effects on humans. This study aimed to evaluate the potential toxicity of A. altissima leaves, using a complex approach towards different organisms. The ecotoxic impact of a crude extract was investigated on seeds germination and brine shrimp lethality. Cytotoxicity was studied in vitro using non-target (haemolysis, liposomal model, fibroblast), and target (cancer cells) assays. Leaf extract at 1000 µg/mL significantly inhibited wheat and tomato germination, while no significant effects were found on parsley germination. A slight stimulatory effect on wheat and tomato germination was found at 125 µg/mL. In a brine shrimp-test, the extract showed a low toxicity at 24 h post-exposure (LC50 = 951.04 ± 28.26 µg/mL), the toxic effects increasing with the exposure time and extract concentration. Leaf extract caused low hematotoxicity. The extract was biocompatible with human gingival fibroblasts. No anti-proliferative effect was found within the concentration range of 10-500 µg/mL on malignant melanoma (MeWo) and hepatocellular carcinoma (HepG2). In a liposomal model-test, the extract proved to possess low capability to alter the eukaryotic cell-mimicking membranes within the tested concentration range. Given the low to moderate toxicity on tested organisms/cells, the A. altissima autumn leaves may find useful applications.

Relevance of amphiphilicity and helicity on the antibacterial action of a histatin 5-derived peptide.

Peptide dhvar4, derived from the active domain of our salivary peptide histatin 5, bears a Phe residue in the middle of its hydrophilic face when folded into an α-helix. We then synthesized an analog with this Phe replaced by Lys and two analogs preserving Phe but bearing two and three α-aminoisobutyric acid (Aib) residues to stabilize the helical structure. The aim of this design was to verify which of the two features is more favorable to the biological activity. We performed a conformational study by means of circular dichroism and nuclear magnetic resonance, made antibacterial tests, and assessed the stability of the peptides in human serum. We observed that amphiphilicity is more important than helix stability, provided a peptide can adopt a helical conformation in a membrane-mimetic environment.

A Study of Hyaluronic Acid's Theoretical Reactivity and of Magnetic Nanoparticles Capped with Hyaluronic Acid.

Hyaluronic acid (HA) has attracted much attention in tumor-targeted drug delivery due to its ability to specifically bind to the CD44 cellular receptor, which is widely expressed on cancer cells. We present HA-capped magnetic nanoparticles (HA-MNPs) obtained via the co-precipitation method, followed by the electrostatic adsorption of HA onto the nanoparticles' surfaces. A theoretical study carried out with the PM3 method evidenced a dipole moment of 3.34 D and negatively charged atom groups able to participate in interactions with nanoparticle surface cations and surrounding water molecules. The ATR-FTIR spectrum evidenced the hyaluronic acid binding to the surface of the ferrophase, ensuring colloidal stability in the water dispersion. To verify the success of the synthesis and stabilization, HA-MNPs were also characterized using other investigation techniques: TEM, EDS, XRD, DSC, TG, NTA, and VSM. The results showed that the HA-MNPs had a mean physical size of 9.05 nm (TEM investigation), a crystallite dimension of about 8.35 nm (XRD investigation), and a magnetic core diameter of about 8.31 nm (VSM investigation). The HA-MNPs exhibited superparamagnetic behavior, with the magnetization curve showing saturation at a high magnetic field and a very small coercive field, corresponding to the net dominance of single-domain magnetic nanoparticles that were not aggregated with reversible magnetizability. These features satisfy the requirement for magnetic nanoparticles with a small size and good dispersibility for long-term stability. We performed some preliminary tests regarding the nanotoxicity in the environment, and some chromosomal aberrations were found to be induced in corn root meristems, especially in the anaphase and metaphase of mitotic cells. Due to their properties, HA-MNPs also seem to be suitable for use in the biomedical field.

Influence of Convective and Vacuum-Type Drying on Quality, Microstructural, Antioxidant and Thermal Properties of Pretreated Boletus edulis Mushrooms.

Freshly harvested Boletus edulis mushrooms are subjected to rapid loss of quality due to the high moisture content and enzymatic activity. Drying time, quality characteristics, microstructural and thermal properties were studied in mushrooms ground to puree subjected to hot air drying (HAD), freeze drying (FD) and centrifugal vacuum drying (CVD). The influence of hot water blanching and UV-C pretreatments was additionally investigated. The rehydration ability of mushroom powders was improved by FD, especially without pretreatment or combined to UV-C exposure. The HAD and CVD, with no pretreatment or combined to UV-C, ensured good preservation of phenolics and antioxidant activity of dried mushrooms. The total difference in color of mushroom pigments extracted in acetone was lower in samples dried by CVD and higher in ones by FD. Blanching before HAD produced whiter product probably due to the reduced polyphenoloxidase activity. Scanning Electron Microscopy (SEM) analysis showed fewer physical changes in FD-samples. Heat-induced structural changes were noticed by Differential Scanning Calorimetry (DSC), Thermogravimetry (TG) and Derivative Thermogravimetry (DTG) analysis, in particular of biopolymers, confirmed by ATR-FTIR analysis. Based on our complex approach, the UV pretreatment of mushrooms could be a better alternative to water blanching. Centrifugal vacuum emerged as a new efficient drying method in terms of bioactive compounds, color and thermal stability, while FD led to better rehydration ability and microstructure.

Impact of Magnetite Nanoparticles Coated with Aspartic Acid on the Growth, Antioxidant Enzymes Activity and Chlorophyll Content of Maize.

In recent decades, magnetite nanoparticles received greater attention in nanobiotechnology due to wide applications. This study presents the influence of the oxidative stress caused by magnetite nanoparticles coated with aspartic acid (A-MNP) of 9.17 nm mean diameter size, on maize (Zea mays) seedlings, in terms of growth, enzymatic activity and chlorophyll content as evaluated in exposed plant tissues. Diluted suspensions of colloidal magnetite nanoparticles stabilized in water were added to the culture medium of maize seeds, such as to equate nanoparticle concentrations varying from 0.55 mg/L to 11 mg/L. The obtained results showed that the growth of maize was stimulated by increasing the level of A-MNPs. Plant samples treated with different concentrations of A-MNP proved increased activities of catalase and peroxidase, and chlorophyll content, as well. The exposure of plants to magnetite nanoparticles may induce oxidative stress, which activates the plant defense/antioxidant mechanisms.

Low-Molecular-Weight Synthetic Antioxidants: Classification, Pharmacological Profile, Effectiveness and Trends.

Mounting research has been performed and published on natural antioxidants, more so than on synthetic ones, as key molecules that control oxidative damage and its pathway to disease. Since the discovery of vitamins, various fully synthetic or natural-identical compounds have been developed as stable small molecules translated into constantly active and completely controlled products which are widely exploited in the food and pharmaceutical industries. There is currently a debate within the literature about their mechanism of action, bioavailability, safety and real benefit for human health. Using a semiquantitative method and eligible criteria of selection, this review aimed to provide a very useful classification of antioxidants and a comprehensive cross-disciplinary description of 32 approved synthetic/natural-identical antioxidants, in terms of regulatory, antioxidant mechanism of action, safety issues, pharmacological properties, effectiveness in human health, timeline and future trends. Enriched interpretation of the data was obtained from summary bibliometrics, useful to portray the "good antioxidant" within the period 1966-2021 and, hopefully, to encourage further research.

Aspartic Acid Stabilized Iron Oxide Nanoparticles for Biomedical Applications.

Aspartic acid stabilized iron oxide nanoparticles (A-IONPs) with globular shape and narrow size distribution were prepared by the co-precipitation method in aqueous medium. A quantum-mechanical approach to aspartic acid optimized structure displayed negative charged sites, relatively high dipole moment, and hydrophilicity, which recommended it for interaction with iron cations and surrounding water electrical dipoles. A-IONPs were characterized by TEM, XRD, ATR-FTIR, EDS, DSC, TG, DLS, NTA, and VSM techniques. Theoretical study carried out by applying Hartree-Fock and density functional algorithms suggested that some aspartic acid properties related to the interaction can develop with nanoparticles and water molecules. The results of experimental investigation showed that the mean value of particle physical diameters was 9.17 ± 2.2 nm according to TEM image analysis, the crystallite size was about 8.9 nm according to XRD data, while the magnetic diameter was about 8.8 nm, as was determined from VSM data interpretation with Langevin's theory. The A-IONP suspension was characterized by zeta-potential of about -11.7 mV, while the NTA investigation revealed a hydrodynamic diameter of 153.9 nm. These results recommend the A-IONP suspension for biomedical applications.

A Review of the Current Knowledge of Thermal Stability of Anthocyanins and Approaches to Their Stabilization to Heat.

Anthocyanins are colored valuable biocompounds, of which extraction increases globally, although functional applications are restrained by their limited environmental stability. Temperature is a critical parameter of food industrial processing that impacts on the food matrix, particularly affecting heat-sensitive compounds such as anthocyanins. Due to the notable scientific progress in the field of thermal stability of anthocyanins, an analytical and synthetic integration of published data is required. This review focuses on the molecular mechanisms and the kinetic parameters of anthocyanin degradation during heating, both in extracts and real food matrices. Several kinetic models (Arrhenius, Eyring, Ball) of anthocyanin degradation were studied. Crude extracts deliver more thermally stable anthocyanins than purified ones. A different anthocyanin behavior pattern within real food products subjected to thermal processing has been observed due to interactions with some nutrients (proteins, polysaccharides). The most recent studies on the stabilization of anthocyanins by linkages to other molecules using classical and innovative methods are summarized. Ensuring appropriate thermal conditions for processing anthocyanin-rich food will allow a rational design for the future development of stable functional products, which retain these bioactive molecules and their functionalities to a great extent.

Kinetic studies on the oxidative stabilization effect of red onion skins anthocyanins extract on parsley (Petroselinum crispum) seed oil.

Cold pressed parsley seed oil gained special interest for the development of new functional foods. Similar to other edible oils it needs protection against oxidation. The present study evaluated the protective effects of anthocyanins extracted from outer skins of red onion on parsley seed oil. The natural extract-oil samples and control have been subjected to storage at 45 °C for 10 days. The results of thermal analysis by DSC showed an optimal concentration of anthocyanins of 3 mL/100 g oil. Kinetic studies using the Ozawa-Flynn-Wall and Kissinger-Akahira-Sunose methods were performed on the sample optimally prepared. The results regarding the activation energy indicated improved stability of parsley seed oil in the presence of anthocyanins. The kinetic studies based on peroxide values also suggested the protective role of red onion anthocyanins. The results are important for the development of natural alternatives useful for the inhibition of oxidation in parsley seed oil.

Conformational properties, membrane interaction, and antibacterial activity of the peptaibiotic chalciporin A: Multitechnique spectroscopic and biophysical investigations on the natural compound and labeled analogs.

In this work, an extensive set of spectroscopic and biophysical techniques (including FT-IR absorption, CD, 2D-NMR, fluorescence, and CW/PELDOR EPR) was used to study the conformational preferences, membrane interaction, and bioactivity properties of the naturally occurring synthetic 14-mer peptaibiotic chalciporin A, characterized by a relatively low (≈20%), uncommon proportion of the strongly helicogenic Aib residue. In addition to the unlabeled peptide, we gained in-depth information from the study of two labeled analogs, characterized by one or two residues of the helicogenic, nitroxyl radical-containing TOAC. All three compounds were prepared using the SPPS methodology, which was carefully modified in the course of the syntheses of TOAC-labeled analogs in view of the poorly reactive α-amino function of this very bulky residue and the specific requirements of its free-radical side chain. Despite its potentially high flexibility, our results point to a predominant, partly amphiphilic, α-helical conformation for this peptaibiotic. Therefore, not surprisingly, we found an effective membrane affinity and a remarkable penetration propensity. However, chalciporin A exhibits a selectivity in its antibacterial activity not in agreement with that typical of the other members of this peptide class.

Reliability of a decision-tree model in predicting occupational lead poisoning in a group of highly exposed workers.

OBJECTIVE: This study aimed to provide the toxicological profile of some lead-exposed workers and obtain a predictive model for lead poisoning. METHODS: Data regarding external and absorbed exposure were collected from 585 subjects employed in ten metallurgical production departments. Airborne lead concentration, blood lead level (BLL), cumulative blood lead index (CBLI), urine delta-aminolevulinic acid (DALA), age, workplace/section, exposure period, and whether reported lead poisoning as occupational disease were examined using ANOVA, and, post-ANOVA, Pearson correlation matrix, PCA (principal component analysis), decision-tree modeling, and logistic modeling. RESULTS: BLL was less sensitive than CBLI in predicting poisoning. Decision-tree modeling highlighted the importance of CBLI ≥1,041 µg.years/dl and air lead concentration ≥0.3 mg/m(3) in the occurrence of occupational poisoning. Age ≥48 years and DALA ≥19.3 mg/L were also factors. CONCLUSIONS: Workers were at risk of poisoning as a result of their long term unacceptable exposure. Decision-tree modeling is potentially useful for risk management. Am. J. Ind. Med. 59:575-582, 2016. © 2016 Wiley Periodicals, Inc.

The effect of ultrasonic pretreatment and sample preparation on the extraction yield of antioxidant compounds and activity of black currant fruits.

The aim of the study was to evaluate the efficiency of an ultrasonic pretreatment at different amplitudes and extraction times, on the content of antioxidant compounds (phenolics, anthocyanins, ascorbic acid) and total antioxidant capacity of black currant fruits. Additionally, the influence of sample preparation (frozen storage/drying) was evaluated. Extraction was performed in 60% ethanol with 0.15% HCl at a solvent-to-sample ratio of 15/1. Our results show that the ultrasonic pretreatment proved particularly useful for the recovery of high amounts of total anthocyanins in freeze-dried samples, ascorbic acid in frozen and oven air-dried samples, and total antioxidant capacity in freeze-dried and oven air-dried samples. The total phenolics content was not significantly influenced by the ultrasonic pretreatment. Freeze-drying increased the content of targeted compounds, to a very high significant level (p < 0.001) for anthocyanins and ascorbic acid. Oven air-drying at 45 °C drastically decreased the ascorbic acid content. Ultrasonication enhanced the extraction yield of black currant valuable compounds in strong relation to the operating conditions.

Solution synthesis, conformational analysis, and antimicrobial activity of three alamethicin F50/5 analogs bearing a trifluoroacetyl label.

We prepared, by solution-phase methods, and fully characterized three analogs of the membrane-active peptaibiotic alamethicin F50/5, bearing a single trifluoroacetyl (Tfa) label at the N-terminus, at position 9 (central region) or at position 19 (C-terminus), and with the three Gln at positions 7, 18, and 19 replaced by Glu(OMe) residues. To add the Tfa label at position 9 or 19, a γ-trifluoroacetylated α,γ-diaminobutyric acid (Dab) residue was incorporated as a replacement for the original Val(9) or Glu(OMe)(19) amino acid. We performed a detailed conformational analysis of the three analogs (using FT-IR absorption, CD, 2D-NMR, and X-ray diffraction), which clearly showed that Tfa labeling does not introduce any dramatic backbone modification in the predominantly α-helical structure of the parent peptaibiotic. The results of an initial solid-state (19)F-NMR study on one of the analogs favor the conclusion that the Tfa group is a very promising reporter for the analysis of peptaibioticmembrane interactions. Finally, we found that the antimicrobial activities of the three newly synthesized analogs depend on the position of the Tfa label in the peptide sequence.

Conventional and ultrasound-assisted extraction of anthocyanins from blackberry and sweet cherry cultivars.

Blackberry and sweet cherry are important plant foods rich in anthocyanins well-known for their pharmacological and antioxidant effects. The aim of the present paper was to comparatively investigate conventional and ultrasound-assisted extraction procedures in order to isolate an enriched crude anthocyanin extract from blackberry (Thornfree cultivar) and sweet cherry (Black Gold cultivar). Hydroethanolic solution and acidified ethanol were used to conventionally extract anthocyanins by a discontinuous process at 4 degreeC for 2/ 24 hours. Added hydrochloric acid in ethanol of different concentrations proved to be more efficient in both type of samples. In the ultrasound-assisted extraction, the highest recovered anthocyanin content in blackberry (107.81 mg 100 g(-1) FM) was obtained with a 10/1 solvent/solid ratio (v/w) at 30 degreeC for 5 minutes, while a 15/1 solvent/solid ratio (v/w) at 30 degreeC for 20 minutes lead to an increased antioxidant capacity as determined by ferric reducing antioxidant power in the extract using 0.1% HCI in 80% ethanol. The optimum conditions obtained for ultrasound-assisted extraction from sweet cherry in 0.1% HCI in 60% ethanol at 30 degree C include a 15/1 solid/solvent ratio (w/v) and 5 minutes for the maximum yield (36.05 mg 100(-1) FM). The final crude anthocyanin extracts may find useful application as dietary supplements, or may be further purified for application as food ingredients.

Spectroscopically labeled peptaibiotics. Synthesis and properties of selected trichogin GA IV analogs bearing a side-chain-monofluorinated aromatic amino acid for (19) F-NMR analysis.

We prepared by solid-phase methods, chromatographically purified, and characterized three analogs of the ten-amino acid-residue, membrane-active, lipopeptaibiotic trichogin GA IV, each containing a single (4-fluorophenyl)alanine in position 3, 7, or 10, where it replaces the hydrophobic residue Leu(3) , Leu(7) , or Ile(10) , respectively. We incorporated the fluorine probe based on the observation that the (19) F-NMR technique has been extensively utilized to analyze peptidemembrane interactions in biological systems. A detailed conformational investigation in solution, including a membrane-mimetic environment, was performed on these compounds using FT-IR absorption, CD, and 2D-NMR, combined with molecular-dynamics calculations. The experimentally observed, mixed 310 /α-helical structures unequivocally show that the principal conformational features of trichogin GA IV are preserved in all three analogs. Analogies and differences between the behavior of the natural lipopeptaibiotic and those of the peptides characterized by the side-chain monofluorinated aromatic amino acid were found in membrane-permeabilization experiments and antimicrobial assays. The results of a preliminary solution (19) F-NMR study support the view that the (19) F label is an excellent reporter for changes in the helical environment of the peptide.

Spectroscopically labeled peptaibiotic analogs: the 4-nitrophenylalanine infrared absorption probe inserted at different positions into trichogin GA IV.

A set of three analogs of the 10-residue, membrane-active lipopeptaibiotic trichogin GA IV, labeled with the promising 4-nitrophenylalanine IR absorption probe for local polarity, was synthesized by the solid-phase methodology, chromatographically purified, and extensively characterized. A single residue modification was inserted near the N-terminus, in the central region, or at the C-terminus. A solution conformational analysis, carried out by FT-IR absorption, CD, and 2D-NMR combined with molecular dynamics calculations, indicates that the mono-labeled analogs maintain the overall helical properties of the parent compound. Membrane permeabilization measurements and antimicrobial tests revealed that they possess membrane-modifying properties and in vitro antibacterial activities analogous to those of the natural lipopeptaibiotic. Our IR absorption and attenuated total reflectance investigations in various environments, from which water was excluded for solubility reasons, showed that the 1350 cm(-1) 4-nitrobenzyl band is a reporter group of rather limited sensitivity.

Partial thioamide scan on the lipopeptaibiotic trichogin GA IV. Effects on folding and bioactivity.

Backbone modification is a common chemical tool to control the conformation of linear peptides and to explore potentially useful effects on their biochemical and biophysical properties. The thioamide, ψ[CS-NH], group is a nearly isosteric structural mimic of the amide (peptide) functionality. In this paper, we describe the solution synthesis, chemical characterization, preferred conformation, and membrane and biological activities of three, carefully selected, peptide analogues of the lipopeptaibiotic [Leu(11)-OMe] trichogin GA IV. In each analogue, a single thioamide replacement was incorporated. Sequence positions near the N-terminus, at the center, and near the C-terminus were investigated. Our results indicate that (i) a thioamide linkage is well tolerated in the overall helical conformation of the [Leu(11)-OMe] lipopeptide analogue and (ii) this backbone modification is compatible with the preservation of its typical membrane leakage and antibiotic properties, although somewhat attenuated.

Synthesis and preliminary conformational analysis of TOAC spin-labeled analogues of the medium-length peptaibiotic tylopeptin B.

A set of analogues of the 14-residue peptaibol tylopeptin B, containing the stable free-radical 4-amino-1-oxyl-2,2,6,6,-tetramethylpiperidine-4-carboxylic acid (TOAC) at one or two selected positions, was synthesized by the solid-phase methodology. A solution conformational analysis performed by FTIR absorption and CD suggests that, in membrane-mimicking solvents, the labeled tylopeptin B analogues preserve the helical propensity of the parent peptide, with a preference for the α-helix or the 3(10) -helix type depending upon the nature of the solvent. In aqueous environment, the spin-labeled analogues present a higher content of helical conformation as a consequence of the strong helix promoter effect of the conformationally constrained TOAC residue. We observed a progressive increase of the quenching effect of the nitroxyl radical on the fluorescence of the N-terminal tryptophan as TOAC replaces the Aib residue at positions 13, 8, and 4, respectively. A membrane permeabilization assay performed on two selected analogues, TOAC(8) - and TOAC(13) -tylopeptin B, showed that the labeled peptides exhibit membrane-modifying properties comparable with those of the natural peptaibiotic. We conclude that our TOAC paramagnetic analogues of tylopeptin B are good models for a detailed ESR investigation of the mechanism of membrane permeabilization induced by medium-length peptaibiotics.

Immunodetection of added glycomacropeptide in milk formulas and milk powders.

The present study aimed the detection of fraudulent manipulation of milk powder with a low cost component--whey powder, by applying the immunochromatographic assay to identify glycomacropeptide. Five commercial milk powder samples of various brands from the national market were analyzed: lactose enriched milk powder type 26, two whole milk powders, vitamin enriched milk powder and full cream milk powder. Our results showed additional whey (1-2%) in 60% of the selected samples after casein removal by precipitation with 20% trichloracetic acid. Another investigated sample--the enriched UHT milk for children aged 4-12 years--proved addition of whey. Other two commercial toddler formula milk powder samples of different brands were used for comparison for the presence of glycomacropeptide. The first sample which was regularly labeled as containing whey protein concentrate was found positive for glycomacropeptide in accordance with the label information, while the second one not containing whey proteins as specified by the product label, was found negative for glycomacropeptide, these two samples being in accordance with the actual legislation.