Synthesis and Characterization of a Biopolymer Pectin/Ethanolic Extract from Olive Mill Wastewater: In Vitro Safety and Efficacy Tests on Skin Wound Healing.

Wound-healing delay is one of the major problems of type 2 diabetes, representing also a clinical emergency in non-healing chronic wounds. Natural antioxidants show interesting wound-healing properties, including those extracted from waste derived from olive oil production. Olive mill wastewater is one of the main by-products of the olive oil-making process, and it is rich in high-value secondary metabolites, mainly hydroxytyrosol. We proposed an eco-friendly extraction method, employing both ultrasound-assisted and Soxhlet techniques and ethanol as a solvent, to recover valuable molecules from Roggianella cv (Olea europea L.) olive mill wastewater, which was further entrapped in a pectin polymer via an enzymatic reaction using porcine pancreatic lipase. Pectin, in combination with other substances, promoted and accelerated wound healing and demonstrated good potential to produce a biomedical conjugate for wound treatment. The antioxidant activity of the extracts and conjugate were evaluated against lipophilic (IC50 equal to 0.152 mg mL-1) and hydrophilic (IC50 equal to 0.0371 mg mL-1) radical species as well as the in vitro cytotoxicity via NRU, h-CLAT, and a wound-healing scratch assay and assessment. The pectin conjugate did not exert hemolytic effects on the peripheral blood, demonstrating interesting wound-healing properties due to its ability to stimulate cell proliferation in a dose-dependent manner.

Overview of the Potential Beneficial Effects of Carotenoids on Consumer Health and Well-Being.

Well-known experimental research demonstrates that oxidative stress is the leading cause of the onset and progression of major human health disorders such as cardiovascular, neurological, metabolic, and cancer diseases. A high concentration of reactive oxygen species (ROS) and nitrogen species leads to damage of proteins, lipids, and DNA associated with susceptibility to chronic human degenerative disorders. Biological and pharmaceutical investigations have recently focused on exploring both oxidative stress and its defense mechanisms to manage health disorders. Therefore, in recent years there has been considerable interest in bioactive food plant compounds as naturally occurring antioxidant sources able to prevent, reverse, and/or reduce susceptibility to chronic disease. To contribute to this research aim, herein, we reviewed the beneficial effects of carotenoids on human health. Carotenoids are bioactive compounds widely existing in natural fruits and vegetables. Increasing research has confirmed that carotenoids have various biological activities, such as antioxidant, anti-tumor, anti-diabetic, anti-aging, and anti-inflammatory activities. This paper presents an overview of the latest research progress on the biochemistry and preventative and therapeutic benefits of carotenoids, particularly lycopene, in promoting human health. This review could be a starting point for improving the research and investigation of carotenoids as possible ingredients of functional health foods and nutraceuticals in the fields of healthy products, cosmetics, medicine, and the chemical industry.

Discrimination of Olive Oil and Extra-Virgin Olive Oil from Other Vegetable Oils by Targeted and Untargeted HRMS Profiling of Phenolic and Triterpenic Compounds Combined with Chemometrics.

Extra-virgin olive oil (EVOO) and virgin olive oil (VOO) are valuable natural products of great economic interest for their producing countries, and therefore, it is necessary to establish methods capable of proving the authenticity of these oils on the market. This work presents a methodology for the discrimination of olive oil and extra-virgin olive oil from other vegetable oils based on targeted and untargeted high-resolution mass spectrometry (HRMS) profiling of phenolic and triterpenic compounds coupled with multivariate statistical analysis of the data. Some phenolic compounds (cinnamic acid, coumaric acids, apigenin, pinocembrin, hydroxytyrosol and maslinic acid), secoiridoids (elenolic acid, ligstroside and oleocanthal) and lignans (pinoresinol and hydroxy and acetoxy derivatives) could be olive oil biomarkers, whereby these compounds are quantified in higher amounts in EVOO compared to other vegetable oils. The principal component analysis (PCA) performed based on the targeted compounds from the oil samples confirmed that cinnamic acid, coumaric acids, apigenin, pinocembrin, hydroxytyrosol and maslinic acid could be considered as tracers for olive oils authentication. The heat map profiles based on the untargeted HRMS data indicate a clear discrimination of the olive oils from the other vegetable oils. The proposed methodology could be extended to the authentication and classification of EVOOs depending on the variety, geographical origin, or adulteration practices.

Overview of the Polyphenols in Salicornia: From Recovery to Health-Promoting Effect.

Nowadays, there has been considerable attention paid toward the recovery of waste plant matrices as possible sources of functional compounds with healthy properties. In this regard, we focus our attention on Salicornia, a halophyte plant that grows abundantly on the coasts of the Mediterranean area. Salicornia is used not only as a seasoned vegetable but also in traditional medicine for its beneficial effects in protecting against diseases such as obesity, diabetes, and cancer. In numerous research studies, Salicornia consumption has been highly suggested due to its high level of bioactive molecules, among which, polyphenols are prevalent. The antioxidant and antiradical activity of polyphenols makes Salicornia a functional food candidate with potential beneficial activities for human health. Therefore, this review provides specific and compiled information for optimizing and developing new extraction processes for the recovery of bioactive compounds from Salicornia; focusing particular attention on polyphenols and their health benefits.

Determination of hydroxytyrosol and tyrosol in human urine after intake of extra virgin olive oil produced with an ultrasounds-based technology.

Extra virgin olive oil (EVOO) is a known source of antioxidants, such as phenolic compounds, useful in the prevention of non-infectious diseases (atherosclerosis, diabetes, cancer, and other diseases). In the present study, EVOO obtained using an innovative ultrasounds-based technology was found richer in total polyphenols, hydroxytyrosol and tyrosol, than EVOO obtained using a conventional mechanical technology. The urinary excretion in humans of hydroxytyrosol and tyrosol, after the administration of ultrasounds and mechanical EVOOs, respectively, was assessed and compared. The analytes were determined in urine samples, collected for 24 h, of six healthy people (3 men and 3 women, age 22-70 years and body mass index <30 kg/m2) who ingested 20 g of oil for six consecutive days. A commercial refined olive oil was also used in the study to determine the baseline excretion levels of the two metabolites. High correlation coefficients (≥0.9311) were found between the amounts of the analytes ingested daily with EVOOs and those determined in the 24-h urines. The results clearly indicated that the EVOO obtained with the ultrasound process was characterized by the highest concentration of biophenols which were consequently available in greater quantities after ingestion, indicating that it represents a high-quality product containing high levels of beneficial compounds such as biophenols readily assimilable by the human body.

Extra Virgin Olive Oil Phenol Extracts Exert Hypocholesterolemic Effects through the Modulation of the LDLR Pathway: In Vitro and Cellular Mechanism of Action Elucidation.

This study was aimed at investigating the hypocholesterolemic effects of extra virgin olive oil (EVOO) phenols and the mechanisms behind the effect. Two phenolic extracts were prepared from EVOO of different cultivars and analyzed using the International Olive Council (IOC) official method for total phenols, a recently validated hydrolytic procedure for total hydroxytyrosol and tyrosol, and 1H-NMR analysis in order to assess their secoiridoid profiles. Both of the extracts inhibited in vitro the 3-hydroxy-3-methylglutaryl co-enzyme A reductase (HMGCoAR) activity in a dose-dependent manner. After the treatment of human hepatic HepG2 cells (25 µg/mL), they increased the low-density lipoprotein (LDL) receptor protein levels through the activation of the sterol regulatory element binding proteins (SREBP)-2 transcription factor, leading to a better ability of HepG2 cells to uptake extracellular LDL molecules with a final hypocholesterolemic effect. Moreover, both of the extracts regulated the intracellular HMGCoAR activity through the increase of its phosphorylation by the activation of AMP-activated protein kinase (AMPK)-pathways. Unlike pravastatin, they did not produce any unfavorable effect on proprotein convertase subtilisin/kexin 9 (PCSK9) protein level. Finally, the fact that extracts with different secoiridoid profiles induce practically the same biological effects suggests that the hydroxytyrosol and tyrosol derivatives may have similar roles in hypocholesterolemic activity.

Optimization of Microwave-Assisted Extraction of Antioxidants from Bamboo Shoots of Phyllostachys pubescens.

Bamboo is a well-known medicinal plant in Southeast Asia that recently has attracted attention for its high polyphenol content and its medical and nutraceutical applications. In this work, polyphenols have been recovered for the first time by microwave-assisted extraction (MAE) from an unusual Italian cultivar of Phyllostachys pubescens bamboo shoots. The effects of three independent variables, such as extraction time, temperature, and solid/liquid ratio, on polyphenol recovery yield were investigated and successfully optimized through the response surface methodology. We demonstrated that MAE is an excellent polyphenols extraction technique from bamboo shoots because the total phenolic content obtained under microwave irradiation optimal conditions (4 min at 105 °C with 6.25 mg/mL ratio) was about eight-fold higher than that obtained with the conventional extraction method. Furthermore, higher total flavonoid content was also obtained under MAE. Consistent with these results, MAE enhanced the extract antioxidant properties with significant improved DPPH, ABTS, and FRAP scavenging ability. Therefore, this innovative extraction process enhances the recovery of biologically active compounds from Phyllostachys pubescens bamboo shoots with a dramatic reduction of time and energy consumption, which paves the way for its industrial application in functional food production.

Effects of Sweet Cherry Polyphenols on Enhanced Osteoclastogenesis Associated With Childhood Obesity.

Childhood obesity is associated with the development of severe comorbidities, such as diabetes, cardiovascular diseases, and increased risk of osteopenia/osteoporosis and fractures. The status of low-grade inflammation associated to obesity can be reversed through an enhanced physical activity and by consumption of food enrich of anti-inflammatory compounds, such as omega-3 fatty acids and polyphenols. The aim of this study was to deepen the mechanisms of bone impairment in obese children and adolescents through the evaluation of the osteoclastogenic potential of peripheral blood mononuclear cells (PBMCs), and the assessment of the serum levels of RANKL and osteoprotegerin (OPG). Furthermore, we aimed to evaluate the in vitro effects of polyphenol cherry extracts on osteoclastogenesis, as possible dietary treatment to improve bone health in obese subjects. High RANKL levels were measured in obese with respect to controls (115.48 ± 35.20 pg/ml vs. 87.18 ± 17.82 pg/ml; p < 0.01), while OPG levels were significantly reduced in obese than controls (378.02 ± 61.15 pg/ml vs. 436.75 ± 95.53 pg/ml, respectively, p < 0.01). Lower Ad-SoS- and BTT Z-scores were measured in obese compared to controls (p < 0.05). A significant elevated number of multinucleated TRAP+ osteoclasts (OCs) were observed in the un-stimulated cultures of obese subjects compared to the controls. Interestingly, obese subjects displayed a higher percentage of CD14+/CD16+ than controls. Furthermore, in the mRNA extracts of obese subjects we detected a 2.5- and 2-fold increase of TNFα and RANKL transcripts compared to controls, respectively. Each extract of sweet cherries determined a dose-dependent reduction in the formation of multinucleated TRAP+ OCs. Consistently, 24 h treatment of obese PBMCs with sweet cherry extracts from the three cultivars resulted in a significant reduction of the expression of TNFα. In conclusion, the bone impairment in obese children and adolescents is sustained by a spontaneous osteoclastogenesis that can be inhibited in vitro by the polyphenol content of sweet cherries. Thus, our study opens future perspectives for the use of sweet cherry extracts, appropriately formulated as nutraceutical food, as preventive in healthy children and therapeutic in obese ones.

Elucidation of the synergistic action of Mentha Piperita essential oil with common antimicrobials.

Mentha piperita L. essential oil (EO) is employed for external use as antipruritic, astringent, rubefacient and antiseptic. Several studies demonstrated its significant antiviral, antifungal and antibacterial properties. The aim of this work is the study of the synergistic effects of M. piperita EO with antibacterials and antifungals that are widely available and currently prescribed in therapies against infections. The observed strong synergy may constitute a potential new approach to counter the increasing phenomenon of multidrug resistant bacteria and fungi. In vitro efficacy of the association M. piperita EO/drugs was evaluated against a large panel of Gram-positive and Gram-negative bacteria and yeast strains. The antimicrobial effects were studied by checkerboard microdilution method. The synergistic effect of M. piperita EO with gentamicin resulted in a strong growth inhibition for all the bacterial species under study. The synergistic effect observed for M. piperita EO and antifungals was less pronounced.

Working towards the development of innovative ultrasound equipment for the extraction of virgin olive oil.

Malaxation has been recognized as one of the most critical points in the mechanical extraction process for virgin olive oil (VOO). It is a low and continuous kneading of olive paste at a carefully monitored temperature. Through this essential technological operation the small droplets of the oil formed during the milling merge into large drops that can be easily separated with a decanter centrifuge. During this technological phase, a complex and necessary bioprocess takes place in order to determine the quality and composition of the final product. The malaxer is a heat exchanger characterized by a low overall heat transfer coefficient because the ratio of surface area to volume is disadvantageous, so it is important to find an innovative technology to improve heat-exchange. As matter of fact, the malaxing step is the only discontinuous phase in a continuous extraction process. In the next future, the essential challenge of VOO industrial plant manufacturing sector is to design and build advanced machines in order to transform the discontinuous malaxing step in a continuous phase and improve the working capacity of the industrial plants. In order to reduce the malaxing time enhancing the quality of the product, two ultrasound-assisted virgin olive oil extraction processes were tested against the traditional method. The sonication treatment was applied on olives submerged in a water bath (before the crushing) and on olive paste (after the crushing). The ultrasound technology provides a reduction of the malaxing duration improving VOO yields and its minor compounds content. Better extractibility and higher minor compounds contents were obtained by sonicating the olives submerged in a water bath than olive paste. After experimental trials the results were employed to suggest innovative scaling up solutions of the process and new applications of ultrasounds in the VOO industry.