Different Strategies to Obtain Corn (Zea mays L.) Germ Extracts with Enhanced Antioxidant Properties

Maize ( Zea mays L.) germs are by-products from the milling industry. The objective of this work was to compare the phenolic and lipophilic antioxidant fractions of yellow and white corn varieties, provided by Corn Valley S.r.l. (Piumbega, Mantova, Italy) and among the raw materials most processed by the company. The phenolic fraction, extracted with ultrasound-assisted extraction, alone and in combination with chemical and enzymatic hydrolyses, was analyzed with high-performance thin-layer chromatography and reversed-phase high-performance liquid chromatography-diode array detector. Among the various extraction techniques used, the combination of sonication and alkaline hydrolysis proved to be an effective method for the extraction of phenolic compounds from yellow and white germs, with the highest ferulic acid concentrations (636.54 ± 3.71 and 569.23 ± 1.69 mg FA/g dried extract, respectively), total phenolic contents (844.5 ± 64.6 and 742.8 ± 15.44 mg gallic acid equivalents/g dried extract, respectively), and the best antioxidant activity (14.33 ± 0.48 and 11.41 ± 1.1 µg/mL, respectively). The lipophilic fraction, extracted using supercritical carbon dioxide was analyzed by gas chromatography-mass spectrometry. The unsaponifiable fractions were found to be 2.41% ± 0.24% in yellow corn and 1.85% ± 0.08% in white corn; β-sitosterol, campesterol, and stigmasterol were identified as the main phytosterols characterizing both lipophilic extracts which showed the most effective antioxidant activity (1.29 ± 0.26 mg/mL and 1.33 ± 0.21 mg/mL, respectively) compared with the control. Finally, the phenolic and lipophilic extracts obtained from maize by-products may be reintroduced into the health-oriented market as extracts enriched of high-added value biomolecules with antioxidant activity both as active molecules and as additives of natural origin

polyphenols from vitis vinifera lambrusco by products leaves from pruning extraction parameters evaluation through design of experiment

Vitis vinifera L. leaves from pruning are by-products of the wine industry and represent an important source of secondary raw material, thanks to their polyphenols content. Optimization of the extraction processes is a key factor for their valorization, and Design of Experiment (DOE) could be a tool to obtain the most performing extract in terms of polyphenols quality/quantity and bioactivity. Vitis vinifera Lambrusco leaves were subjected to ultrasound-assisted extractions guided by a 23 factorial design. Three independent parameters (% solvent, time of extraction, and solvent:solid ratio) were considered to evaluate the extraction process by analyzing the extraction yield, the total phenolic content (Folin-Ciocalteu assay), and the antioxidant capacity (DPPH assay). Moreover, the content of the main molecules was identified and quantified by reversed-phase high-performance liquid chromatography coupled with diode array detection and mass spectrometry. The DOE highlighted the best extraction conditions that showed slight changes considering the different evaluating parameters. The highest extraction yield was obtained by extraction with 100% water, 60 minutes of extraction time, and 30:1 solvent:solid ratio, but it was neither the richest in polyphenols nor antioxidant capacity. The latter 2 characteristics were associated with the extraction performed using 50% ethanol, 35 minutes of extraction time, and a 20:1 solvent:solid ratio. That extract also exhibited the highest quantity of flavonols

Antimicrobial peptides active in in vitro models of endodontic bacterial infections modulate inflammation in human cardiac fibroblasts

Endodontic and periodontal disease are conditions of infectious origin that can lead to tooth loss or develop into systemic hyperinflammation, which may be associated with a wide variety of diseases, including cardiovascular. Endodontic and periodontal treatment often relies on antibiotics. Since new antimicrobial resistances are a major threat, the use of standard antibiotics is not recommended when the infection is only local. Antimicrobial peptides were recently demonstrated to be valid alternatives for dental treatments. The antimicrobial peptide M33D is a tetrabranched peptide active against Gram-negative and Gram-positive bacteria. It has a long life, unusual for peptides, because its branched form provides resistance to proteases. Here the efficacy of M33D and of its analog M33i/l as antibiotics for local use in dentistry was evaluated. M33D and M33i/l were active against reference strains and multidrug-resistant clinical isolates of Gram-negative and Gram-positive species. Their minimum inhibitory concentration against different strains of dental interest was between 0.4 and 6.0 mu M. Both peptides acted rapidly on bacteria, impairing membrane function. They also disrupted biofilm effectively. Disinfection of the root canal is crucial for endodontic treatments. M33D and M33i/l reduced E. faecalis colonies to one-twentieth in a dentin slices model reproducing root canal irrigation. They both captured and neutralized lipopolysaccharide (LPS), a bacterial toxin responsible for inflammation. The release of IL-1 beta and TNF alpha by LPS-stimulated murine macrophages was reduced by both peptides. Human cardiac fibroblasts respond to different insults with the release of proinflammatory cytokines, and consequently, they are considered directly involved in atherogenic cardiovascular processes, including those triggered by infections. The presence of M33D and M33i/l at MIC concentration reduced IL6 release from LPS- stimulated human cardiac fibroblasts, hence proving to be promising in preventing bacteria-induced atherogenesis. The two peptides showed low toxicity to mammalian cells, with an EC50 one order of magnitude higher than the average MIC and low hemolytic activity. The development of antimicrobial peptides for dental irrigations and medication is a very promising new field of research that will provide tools to fight dental infections and their severe consequences, while at the same time protecting standard antibiotics from new outbreaks of antimicrobial resistance. © 2022 by the authors

Biological activity of new bioactive steroids deriving from biotransformation of cortisone

Cortisone is a metabolite belonging to the corticosteroid class that is used pharmaceutically directly as a drug or prodrug. In addition to its large consumption, its use is linked to several side effects, so pharmaceutical research aims to develop effective drugs with low or no side effects, alternative compounds to cortisone are part of an active investment in ongoing research on drug discovery. Since biotransformation can be considered a source of new molecules with potential therapeutic use, the present work focuses on a preliminary in vitro study aimed at evaluating the mutagenic, anti-inflammatory, antioxidant and neuroprotective activity of SCA and SCB molecules obtained from the biotransformation of cortisone using Rh. Rhodnii strain DSM 43960. The results obtained are very encouraging due to the safety of biotransformed compounds with reference to genotoxicity checked by Ames test, to the very high antioxidant capacity and to the anti-inflammatory activity. In fact, thecompounds inhibited both the TNF alpha-stimulated expression and secretion of NFkB target cytokines, and COX activity, and can activate the glucocorticoid receptor. Finally SCA and SCB exhibited neuroprotective properties

Biological and chemo-diverse characterization of Amazonian (Ecuador) Citrus petitgrains

Six Amazonian petitgrains samples from C. nobilis Lour., C. aurantium L., C. limon L. and mixture of Citrus spp.(Rutaceae), named CN, CA, CL1, CL2, C1 and C2, were chemically characterized by GC-MS and 13C NMR and evaluated for antioxidant acitivity (DPPH and b-carotene bleaching tests), for antimicrobial properties (disk diffusion method) and for antifungal capacity (agar vapour assay). CN, C1, C2 samples evidenced the most interesting results: CN (g-terpinene/linalool chemotype: 14.3%/41.6%, with a considerable amount of thymol: 9.0%), and C1 (linalool, 18.3%; sabinene, 11.6%; thymol, 5.5%), showed relevant antioxidant activity with both DPPH (IC50=3.52 and 5.48 mg/ml, respectively) and b-carotene (IC50=0.387and 0.491 mg/ml, respectively). Antibacterial properties of CN and C1 against P. mirabilis (MIC=0.61 mg/ml for both)and B. subtilis (MIC=0.61 and 0.44 mg/ml, respectively) were most probably due to thymol.C2 (geranial: 34.7%, neral: 33.1%) evidenced a valuable bioactivity against Candida albicans (MIC=0.44 mg/ml).The 50% growth inhibition (IC50) of the dermatophytes T. mentagrophytes and N. cajetani was reached with amounts ofC1, C2 and CN less than 4 ml/plate. Bioactivity of Amazonian Citrus spp. CN, C1 and C2 essential oils suggests their potential use as food preservatives or additives in cosmeceuticals as preventive against dermatophytic fungal infections

Caratterizzazione chimica e biologica di oli essenziali dell’Amazzonia orientale e definizione di un loro profilo applicativo in un contesto salutistico

L’attività di ricerca svolta in questa tesi di dottorato è stata incentrata sulla valorizzazione della biodiversità della foresta amazzonica tramite lo studio di piante aromatiche dell’Ecuador. Tali piante sono particolarmente ricche di oli essenziali, metaboliti secondari costituiti principalmente da molecole volatili odorose, note per essere spesso biologicamente attive. Proprio per via della loro fragranza, gli oli essenziali sono sovente contemplati nelle tradizioni etnomediche delle popolazioni indigene. Questo studio ha come punto di partenza le informazioni della medicina tradizionale con lo scopo di investigare piante poco conosciute potenzialmente interessanti per la salute dell’uomo (in campo farmaceutico, cosmetico ed alimentare). Dopo un’accurata ricerca in letteratura, sono state selezionate per essere studiate le seguenti piante: Chenopodium ambrosioides, Dacryodes peruviana, Piper carpunya, Schinus molle, Tagetes minuta, Endlicheria klugii, Ocotea cernua, Cymbopogon citratus, Ocimum micranthum e Ocotea quixos. Queste specie vegetali fanno parte di differenti famiglie proprio per abbracciare una maggior biodiversità possibile. Grazie ad una collaborazione con l’Universidad Politecnica Salesiana (Quito, Ecuador) e con l’Universidad Estatal Amazonica (Puyo, Ecuador), sono state distillate le droghe fresche delle piante ottenendone gli oli essenziali per arrivare poi ad un “fingerprinting” chimico e biologico. Gli oli essenziali sono stati caratterizzati chimicamente tramite gascromatografia accoppiata alla massa (GC-MS) e risonanza magnetica nucleare (NMR) per una determinazione qualitativa e quantitativa dei composti in essi presenti e successivamente sono stati testati per determinare diverse attività biologiche. Si è partiti dallo studio dell’attività antiossidante mediante il saggio del DPPH, dove l’olio essenziale di O. micranthum ha confermato eccellenti risultati. Sono stati eseguiti test di attività antimicrobica con batteri e funghi sia patogeni per l’uomo che fitopatogeni ed è stata approfondita la loro attività di sinergismo con farmaci di sintesi. Particolarmente interessante si è rilevata l’attività su funghi dermatofiti dove molti dei campioni sono risultati efficaci e il sinergisno con il fluconazolo degli oli essenziali di P. carpunya e S. molle nei confronti di vari ceppi di Candida spp. Sono stati effettuati anche esperimenti di attività antinfiammatoria, nei quali i risultati migliori si sono ottenuti con l’olio essenziale di C. ambrosioides. Per verificare invece la sicurezza per l’uomo sono stati eseguiti test di attività mutagena, dove tutti gli oli essenziali presi in considerazione sono risultati negativi. Infine, è stata realizzata una formulazione cosmentica contenente l’olio essenziale di O. micranthum, ossia quello che si è mostrato più attivo. Lo scopo era quello di trovare un’applicazione per i campioni più promettenti al fine di valorizzare la biodiversità della foresta Amazzonica.The research activities performed in this PhD thesis focuses to promotion of Amazonian rainforest biodiversity by studying aromatic plants of Ecuador. Aromatics plant are reach of essential oils, secondary metabolites consisting mainly in fragrant volatile molecules known to be usually biologically active. Just for their fragrance essential oils are often known by ethnomedicine of indigenous populations. This study starts from information of traditional medicine to investigate poorly known plants potentially interesting for human health (in pharmaceutical, cosmetic and food environment). After careful literature research were choosen these plants Chenopodium ambrosioides, Dacryodes peruviana, Piper carpunya, Schinus molle, Tagetes minuta, Endlicheria klugii, Ocotea cernua, Cymbopogon citratus, Ocimum micranthum and Ocotea quixos. All of different families to consider greater biodiversity possible. Collaborating with the Universidad Politecnica Salesiana (Quito, Ecuador) and with Universidad Estatal Amazonica (Puyo, Ecuador), fresh crude drugs have been distilled to obtain essential oils (EOs) for a phytochemical and biological fingerprinting. The EOs have been chemically characterized through GC-MS and NMR for qualitative and quantitative determination of the compounds and tested for various biological activities. Was performed antioxidant activity with DPPH assay, where O. micranthum essential oil confirmed excellent results. Was performed antimicrobial assays for human or plant pathogens bacteria and fungi, and synergistic studies between essential oil and synthetic drugs where essential oils have been particular efficient against dermatophyte and P. carpunya and S. molle essential oils shown good synergism with fluconazole in Candida spp. Strains. Were conducted antinflamatory tests in which C. ambrosioides essential oil gave best results and mutagenic experiments to verify human safety that excluded potential mutagenic activities. Finally was made a cosmetics formulation with O. micranthum, the more active essential oil. The aims was precisely to find an application for best samples to increase in value the Amazonian biodiversity

Cytotoxic Effect and TLC Bioautography-Guided Approach to Detect Health Properties of Amazonian Hedyosmum sprucei Essential Oil

Bioautography has been used as rapid and easy strategy to detect and identify bioactive fractions/molecules in the never before investigated Hedyosmum sprucei Solms (Chloranthaceae) essential oil (EO). The antioxidant activity, performed through DPPH bioautographic assay and spectrophotometric evaluations (IC50 = 230 ± 10 g/mL), seemed to be mainly due to - cadinol and -muurolol. (HP)TLC bioautography, focused on antimicrobial capacities, pointed out -cadinol, -muurolol, - muurolol, caryophyllene oxide, and methyleugenol as the most effective compounds against Staphylococcus aureus, considered as testing strain. Moreover, the microdilution method, assessed among a wide panel of microorganisms, revealed Listeria grayi and Staphylococcus aureus as the most sensitive among human tested strains and Clavibacter michiganensis among phytopathogens. GCMS chemical profile showed that bioactive molecules represented only a small quantity of the whole EO: germacrene D (23.16%), -caryophyllene (15.53%), -cadinene (5.50%), -copaene (5.08%), and -phellandrene (3.48%) were the main compounds, highlighting an uncommon composition among the genus Hedyosmum. Finally, H. sprucei EO was checked for cytotoxic potential against A549 (lung cancer) and MCF-7 (breast cancer) cell lines showing promising cytotoxic effects against both cell lines after 48 h (IC50 A549 = 44.05 ± 2.35 g/mL; IC50 MCF-7 = 32.76 ± 4.92 g/mL) and 72 h (IC50 A549 = 43.55 ± 2.80 g/mL; IC50 MCF-7 = 33.64 ± 0.43 g/mL

Influence of microbrush on efficacy of bonding into root canals

PURPOSE: Recently, several 'one-bottle' systems have been introduced for bonding into root canals. This study evaluated the influence of two brushes used as carriers of a primer-adhesive solution in the formation of resin tags, adhesive lateral branches and hybrid layer when used to bond fiber posts. MATERIALS AND METHODS: Twenty root canal treated teeth, extracted for periodontal reasons, were selected for this study. The samples were randomly divided into two groups of 10 samples each: Group 1: Microbrush + One Step + Dual Link resin cement; Group 2: One Step + Dual Link resin cement. The adhesive system and resin cement were used strictly following manufacturers' instructions. The priming-adhesive solution of the 'one-bottle' system was light-cured before placing the resin cement and the post. Twenty translucent fiber posts were used. Then, root samples were processed for SEM observation. RESULTS: The adhesive system showed resin dentin interdiffusion zone (RDIZ), resin tag and adhesive lateral branch formation. Microscopic examination of restored interfaces of Group 1 showed a higher % (P < 0.05) of RDIZ than those found in samples of Group 2. In Group 2 samples RDIZ morphology was well detectable and uniform in the first two thirds of root canals while in the apical third the RDIZ was not uniformly present while in Group 1 RDIZ was visible at the apical third too. No statistically significant differences were found among the two groups coronally and at the middle third, while the apical third of Group 1 showed significantly more resin tags than in Group 2. The characteristic reverse cone shape of resin tags was always noted in the coronal and middle third of the root canals of both groups and at the apical third of Group 1. In the apical third of Group 2 root canals, the resin tags showed a less uniform morphology and a shorter length than those found in the other observed thirds