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Abstract Photosynthetic organisms, fungi, and animals comprise distinct pathways for vitamin C biosynthesis. Besides this diversity, the final biosynthetic step consistently involves an oxidation reaction carried out by the aldonolactone oxidoreductases. Here, we study the origin and evolution of the diversified activities and substrate preferences featured by these flavoenzymes using molecular phylogeny, kinetics, mutagenesis, and crystallographic experiments. We find clear evidence that they share a common ancestor. A flavin-interacting amino acid modulates the reactivity with the electron acceptors, including oxygen, and determines whether an enzyme functions as an oxidase or a dehydrogenase. We show that a few side chains in the catalytic cavity impart the reaction stereoselectivity. Ancestral sequence reconstruction outlines how these critical positions were affixed to specific amino acids along the evolution of the major eukaryotic clades. During Eukarya evolution, the aldonolactone oxidoreductases adapted to the varying metabolic demands while retaining their overarching vitamin C-generating function.

Recently, as the interest in cannabidiol (CBD) has grown due to its therapeutic potential, e-cigarette liquids containing CBD have proliferated on the market. Typically, e-liquids contain variable concentrations of CBD (from 2 mg∙mL−1 to 20 mg∙mL−1) in propylene glycol or 70:30 propylene glycol:glycerol mixture and are eventually flavored with food-grade flavors. In this work, carried out by a GC-MS analysis of the condensed smoke produced by a real e-cig, we have demonstrated the actual amount of CBD that can survive the smoking process, and we found that negligible amounts of THCs are formed during the smoking process (i.e., the amount formed was 9-THC is around 2.5 mg (smoked or ingested per os), it is reasonable to conclude that accidental THC intoxication is unlikely, which is a very important issue from a forensic point of view, as in some court cases the use of e-cig liquids containing CBD has been argued as being the source of THC intoxication. Furthermore, all the other cannabinoids considered in this study and potentially derived from CBD thermal degradation have concentrations below the instrumental LOD.

Abstract Background Waiting lists that continue to grow and the lack of organs available for transplantation necessitate the use of marginal livers, such as fatty livers. Since steatotic livers are more susceptible to damage from ischemia and reperfusion, it was investigated whether fatty livers with different lipidomic profiles show a different outcome when subjected to long-term cold storage preservation. Methods Eight-week-old male Wistar rats fed for 2 weeks by a methionine-choline-deficient (MCD) diet or control diet were employed in this study. Livers were preserved in a University of Wisconsin (UW) solution at 4 °C for 6, 12 or 24 h and, after washout, reperfused for 2 h with a Krebs-Henseleit buffer at 37 °C. Hepatic enzyme release, bile production, O2-uptake, and portal venous pressure (PVP) were evaluated. The liver fatty acid profile was evaluated by a gas chromatography-mass spectrometry (GC/MS). Results MCD rats showed higher LDH and AST levels with respect to the control group. When comparing MCD livers preserved for 6, 12 or 24 h, no differences in enzyme release were found during both the washout or the reperfusion period. The same trend occurred for O2-uptake, PVP, and bile flow. A general decrease in SFA and MUFA, except for oleic acid, and a decrease in PUFA, except for arachidonic, eicosadienoic, and docosahexanaeoic acids, were found in MCD rats when compared with control rats. Moreover, the ratio between SFA and the various types of unsaturated fatty acids (UFA) was significantly lower in MCD rats. Conclusions Although prolonged cold ischemia negatively affects the graft outcome, our data suggest that the quality of lipid constituents could influence liver injury during cold storage: the lack of an increased hepatic injury in MCD may be justified by low SFA, which likely reduces the deleterious tendency toward lipid crystallization occurring under cold ischemia.

The autofluorescence of specific fatty acids, retinoids, and bilirubin in crude serum can reflect changes in liver functional engagement in maintaining systemic metabolic homeostasis. The role of these fluorophores as intrinsic biomarkers of pharmacological actions has been investigated here in rats administered with obeticholic acid (OCA), a Farnesoid-X Receptor (FXR) agonist, proven to counteract the increase of serum bilirubin in hepatic ischemia/reperfusion (I/R) injury. Fluorescence spectroscopy has been applied to an assay serum collected from rats submitted to liver I/R (60/60 min ± OCA administration). The I/R group showed changes in the amplitude and profiles of emission spectra excited at 310 or 366 nm, indicating remarkable alterations in the retinoid and fluorescing fatty acid balance, with a particular increase in arachidonic acid. The I/R group also showed an increase in bilirubin AF, detected in the excitation spectra recorded at 570 nm. OCA greatly reversed the effects observed in the I/R group, confirmed by the biochemical analysis of bilirubin and fatty acids. These results are consistent with a relationship between OCA anti-inflammatory effects and the acknowledged roles of fatty acids as precursors of signaling agents mediating damaging responses to harmful stimuli, supporting serum autofluorescence analysis as a possible direct, real-time, cost-effective tool for pharmacological investigations.

The stability of cannabidiol (CBD) under thermal (heating, microwave irradiation, presence of Bronsted and Lewis acids) and photochemical conditions has been tested in organic solvents, including glycerol and propylene glycol that are traditionally employed in the preparation of CBD-containing formulation (e-liquid) for e-cigarettes.The results obtained in the present investigation point out the instability of CBD in the realistic scenario of pharmaceutical preparations and commercial products (e.g., e-liquids). In simulated e-liquid, among the by-products, the psychotropic THCs were found in negligible amounts (

Retinol is transported in vertebrate plasma bound to a protein called retinol-binding protein (RBP4) so far believed to be specific for the vitamin. When the protein is saturated with retinol it binds tightly to another plasma protein, transthyretin while when not saturated with retinol it does not bind to TTR (Goodman, 1984). The X-ray structures of human RBP4, holo and devoid of retinol in its binding site are known to resolutions of 2.0 and 2.5 Å (Cowan et al., 1990; Zanotti et al., 1993) [2,3]. We have shown that RBP4 is not specific for retinol but it is also found in plasma, urine and amniotic fluid bound to fatty acids. Here we present 1.5 Å resolution crystal data on human plasma retinol-binding protein bound to retinol and fatty acids. These are the highest resolution data available in the Protein Data Bank for this protein.For further details and experimental findings please refer to the article “ Human plasma retinol-binding protein (RBP4) is also a fatty acid-binding protein” (Perduca et al., 2018) [4].

The reuse of byproducts from agricultural and food industries represents the key factor in a circular economy, whose interest has grown in the last two decades. Thus, the extraction of bioactives from agro-industrial byproducts is a potential source of valuable molecules. The aim of this work was to investigate the in vitro capacity of byproducts from a new Italian corn variety, named Moradyn, to inhibit the accumulation of advanced glycation end products (AGEs) involved in several chronic age-related disorders. In addition, the hypoglycemic effect of Moradyn was tested by in vitro enzymatic systems. A Moradyn phytocomplex and its purified anthocyanin fraction were able to inhibit fructosamine formation and exhibited antiglycative properties when tested using BSA-sugars and BSA-methylglyoxal assays. These properties could be attributed to the polyphenols, mainly anthocyanins and flavonols, detected by RP-HPLC-DAD-ESI-MSn. Finally, a Moradyn phytocomplex was submitted to a simulated in vitro digestion process to study its bioaccessibility. Moradyn could be considered as a promising food ingredient in the context of typical type 2 diabetes risk factors and the study will continue in the optimization of the ideal formulation to preserve its bioactivities from digestion.

Hydrocarbons can have very harmful effects on organisms and the environment, and conventional techniques for their removal are expensive and require the use of chemicals and long-term actions. Trichoderma is an ascomycete genus known to be active on different recalcitrant substrates, since it can produce a set of nonspecific extracellular enzymes generally involved in the degradation of lignin. However, the literature concerning the use of Trichoderma to degrade hydrocarbons is still limited. In this work we aimed to investigate the ability of Trichoderma to exploit used engine oil as its sole carbon source for prospective bioremediation of contaminated substrates. Four different strains belonging to Trichoderma asperellum and Trichoderma harzianum species were tested. The fungi were inoculated in direct contact with used engine oil, and after 45 days the samples were analyzed by gas chromatography/mass spectrometry (GC/MS). The results showed that all strains (except Trichoderma asperellum F1020) significantly changed the oil composition, decreasing the aromatic fraction in favor of the aliphatic one. T. harzianum F26, especially, showed a significant reduction of the BTEX (benzene, toluene, ethylbenzene, and the three xylene isomers) and alkylbenzenes fraction and an increase in short-chain aliphatics C1–C20. Enzymatic tests for laccase and peroxidase were also carried out, demonstrating that every strain seems to express a different mode of action.

Hericium erinaceus is a medicinal mushroom that contains many molecules promising a plethora of therapeutic properties. In this study, the strain H.e.2 (MicUNIPV, University of Pavia, Italy) was isolated from a sporophore collected in Tuscany (Italy). Mycelium, primordium, and wild type and cultivated sporophores were analyzed by HPLC-UV-ESI/MS. Erinacine A in the mycelium and hericenones C and D in the sporophores were quantified by comparison with their standard molecules. For the first time, H. erinaceus primordium was also investigated for the presence of these molecules. Comparing with the literature data, hericenes, molecules structurally similar to hericenones, were present in all our samples. The highest contents of hericenones C and D were detected in cultivated sporophores, compared to the wild type. The comparison of these data with those of another Italian H. erinaceus strain (H.e.1 MicUNIPV) was discussed. The results led us to select H. erinaceus strains more suitable for mycelium production or sporophore cultivation to obtain extracts with a higher content of bioactive compounds. This work provides a further step towards standardizing the procedures in the development of dietary supplements made from mushrooms.

Multiple myeloma (MM) belongs to hematological cancers and its incidence is increasing worldwide. Despite recent advances in its therapy, MM still causes many deaths every year. In fact, current therapies sometimes fail and are associated with severe adverse effects, including neurotoxicity. As a part of our ongoing efforts to discover new potential therapies against MM, we prepared Hibiscus sabdariffa extracts obtained by a microwave-assisted solvent extraction and investigate their activity by in vitro assays on the RPMI-8226 cell line. The bioguided fractionation of the crude ethanolic extract allowed the identification of HsFC as the most effective extract. We assessed cell viability (MTT and Tripan blue test), cell migration (Boyden chamber assay), and neurotoxicity (DRG neurotoxicity assay). The promising results prompted us to further fractionate HsFC and we obtained two molecules effective against RPMI-8226 cells without neurotoxic effects at their active concentrations. Moreover, both compounds are able to significantly reduce cell migration.

Hydrodeaminated and monodeuterated aromatics were obtained via a visible-light driven reaction of arylazo sulfones. Deuteration occurs efficiently in deuterated media such as isopropanol-d8 or in THF-d8/water mixtures and exhibits a high tolerance to the nature and the position of the aromatic substituents.

Plastic material is used for a wide variety of commercial packaging due to being inexpensive, lightweight, and due to its resistance. In pharmaceutics, container-content compatibility studies are required for product authorization. Many guidelines and publications are available; however, the information is often only related to the raw materials used to produce packaging. During the manufacturing process, substances can be added to improve the product characteristics and performance, resulting in a processed material that is considerably different from the unprocessed material. In this study, the mechanical properties of low-density polyethylene (LDPE) and linear low-density polyethylene (LLDPE) specimens fabricated according to standard ISO 527 and specimens fabricated with the same materials, but obtained from final packaging, were evaluated. Furthermore, we examined the interaction between a semisolid formulation and LLDPE and LDPE as a final packaging, by subjecting two samples to accelerated degradation testing. Then, mechanical properties and volatile organic extractable were evaluated. Simulated solar radiation did not induce changes in the packaging mechanical properties and no extracts were detectable. The thermal shock strongly influenced the mechanical behavior, and interactions between packaging contents were identified. The present work underlines the difference between analyzing the standard ISO specimens versus samples obtained from final packaging in order to evaluate the packaging under real use conditions. An evaluation on the final packaging, instead on standard specimens, can provide information about the plastic material after the manufacturing process and the interaction between packaging and content.

Sunscreens play a fundamental role in skin cancer prevention and in protection against photo-aging. UV filters are often photo-unstable, especially in relation to their vehicles and, being lipophilic substances, they are able to interact with plastic packaging. Finally, UV filter stability can be significantly affected by the routine use of the product at high temperatures. This work aims to study the stability of sunscreen formulations in polyethylene packaging. Butyl methoxydibenzoylmethane and octocrylene, both in a free form and as encapsulated filters were chosen as UV filters. Stability evaluations were performed both in the packaging and on the formulations. Moreover, a further two non-destructive techniques, near-infrared (NIR) spectroscopy and a multiple light scattering technique, were also used to evaluate the stability of the formulation. Results demonstrated clearly that all of the pack underwent significant changes in its elastic/plastic behavior and in external color after solar irradiation. From the evaluation of the extractable profile of untreated and treated packaging material an absorption of 2-phenoxyethanol and octocrylene were shown. In conclusion, the results highlighted clearly that a reduction of the UV filter in the formulation packed in high-density polyethylene/low-density polyethylene (HDPE/LDPE) material can occur over time, reducing the protective effect of the product when applied to the skin.

In the Regulation 1223/2009, evaluation of packaging has become mandatory to assure cosmetic product safety. In fact, the safety assessment of a cosmetic product can be successfully carried out only if the hazard deriving from the use of the designed packaging for the specific product is correctly evaluated. Despite the law requirement, there is too little information about the chemical-physical characteristics of finished packaging and the possible interactions between formulation and packaging; furthermore, different from food packaging, the cosmetic packaging is not regulated and, to date, appropriate guidelines are still missing. The aim of this work was to propose a practical approach to investigate commercial polymeric containers used in cosmetic field, especially through mechanical properties’ evaluation, from a safety point of view. First of all, it is essential to obtain complete information about raw materials. Subsequently, using an appropriate full factorial experimental design, it is possible to investigate the variables, like polymeric density, treatment, or type of formulation involved in changes to packaging properties or in formulation-packaging interaction. The variation of these properties can greatly affect cosmetic safety. In particular, mechanical properties can be used as an indicator of pack performances and safety. As an example, containers made of two types of polyethylene with different density, low-density polyethylene (LDPE) and high-density polyethylene (HDPE), are investigated. Regarding the substances potentially extractable from the packaging, in this work the headspace solid-phase microextraction method (HSSPME) was used because this technique was reported in the literature as suitable to detect extractables from the polymeric material here employed.

In this study, we report the synthesis and characterization of three novel “push–pull” chromophores, in which multiple phenylenevinylene units are endcapped by ferrocene as donor units and malonate moieties as acceptor units. These chromophores have spectroscopic and electrochemical characteristics which consistently change according to the extension of the conjugated bridge, thus to the variation of the HOMO–LUMO band gap. The 1,3-dicarbonyl units, directly incorporated into the conjugated molecular structures, are able to coordinate Lewis acid-like cations, such as lanthanides, as confirmed by UV/Vis, 1H NMR and cyclic voltammetry studies. The UV/Vis spectroscopic response upon complexation with Sc3+or Eu3+as the triflate salts is rather unselective and nonlinear in going from the least to the most π-extended chromophore. Binding studies in MeCN, analyzed viaequilibrium-restricted factor analysis, give values between log Ka= 1.21 and 3.07 and affirm a 1 : 1 stoichiometry of the host:guest complexes in all cases. On the other hand, cyclic voltammetry reveals a selectivity in the response to Sc3+coordination over Ln3+(Eu3+, but also Lu3+and Er3+were tested) for the two shorter chromophores, whereas the ligand with the longest π-bridge is able to sense Er3+(ΔE1/2complexed/uncomplexed chromophore = 20 mV) selectively over the other lanthanides. [ABSTRACT FROM AUTHOR]