Barbara Albertini

Effect of fermentation and drying on cocoa polyphenols.

Cocoa seed polyphenols have demonstrated interesting beneficial effects in humans. Most polyphenols contained in fresh seeds are chemically modified during fermentation, drying, and cocoa powder or chocolate production. The improvement of these procedures to obtain a high-polyphenol-content cocoa is highly desirable. To this aim, a field investigation on the effect of fermentation and natural drying on fine flavor National cocoa (cacao Nacional) was performed. Cocoa seeds were fermented for 6 days and, every day, samples were sun-dried and analyzed for polyphenol content and antioxidant power. During the first 2 days of fermentation, Folin-Ciocalteu and FRAP tests evidenced a significant reduction of polyphenol content and antioxidant capacity, respectively. Changes during the following days of fermentation were less significant. Epicatechin, the most studied member of the catechin family, followed a similar pathway of degradation. Data confirmed the high impact of fermentation and drying on cocoa seed polyphenols. Fermentation and drying are, on the one hand, necessary to obtain cocoa flavor and palatability but, on the other hand, are responsible for greatly compromising polyphenol content. To obtain high-polyphenol-content cocoa, the existing fermentation, drying, and manufacturing protocols should be scientifically reviewed to understand and modify the critical steps.

Antioxidant activity of phenolic extracts from different cultivars of Italian onion (Allium cepa) and relative human immune cell proliferative induction.

Abstract Context: The total antioxidant activity (TAC) may vary considerably between onion cultivars. Immunological effects of onion phenolic compounds are still underestimated. Objective: The objective of this study is to determine the total phenol content (TPC) and the relative TAC of three Allium cepa L. (Liliaceae) onion cultivars cultivated in Cannara (Italy): Rossa di Toscana, Borettana di Rovato, and Dorata di Parma, and to evaluate the phenol extracts ability to induce human immune cell proliferation. Materials and methods: TPC was determined by the Folin–Ciocalteu method, TAC with FRAP, TEAC/ABTS, and DPPH methods. Peripheral blood mononuclear cells from healthy human donors were incubated for 24 h at 37 °C with 1 ng/mL of phenolic extract in PBS, immunostained, and then analyzed by 4-color flow cytometry for the phenotypic characterization of T helper cells (CD4+ cells), cytotoxic T lymphocytes (CD8+ cells), T regulatory cells (CD25high CD4+ cells), and natural killer cells/monocytes (CD16+ cells). Results: Rossa di Toscana displayed the highest TPC (6.61 ± 0.87 mg GA equivalents/g onion bulb DW) and the highest TAC with the experienced methods: FRAP, 9.19 ± 2.54 μmol Trolox equivalents/g onion bulb DW; TEAC/ABTS, 21.31 ± 0.41 μmol Trolox equivalents/g onion bulb DW; DPPH, 22.90 ± 0.01 μmol Trolox equivalents/g onion bulb DW. Incubation with Rossa di Toscana extract determined an increase in the frequency of the antitumor/anti-infection NK CD16+ immune cells (23.0 ± 0.4%). Discussion and conclusions: Content of health-promoting phenols and the deriving antioxidant and immunostimulating activity vary considerably among the investigated cultivars. Rossa di Toscana can be considered as a potential functional food.

Artificial apolipoprotein corona enables nanoparticle brain targeting

Many potential therapeutic compounds for brain diseases fail to reach their molecular targets due to the impermeability of the blood-brain barrier, limiting their clinical development. Nanotechnology-based approaches might improve compounds pharmacokinetics by enhancing binding to the cerebrovascular endothelium and translocation into the brain. Adsorption of apolipoprotein E4 onto polysorbate 80-stabilized nanoparticles to produce a protein corona allows the specific targeting of cerebrovascular endothelium. This strategy increased nanoparticle translocation into brain parenchyma, and improved brain nanoparticle accumulation 3-fold compared to undecorated particles (119.8 vs 40.5 picomoles). Apolipoprotein decorated nanoparticles have high clinical translational potential and may improve the development of nanotechnology-based medicine for a variety of neurological diseases.

Nanoparticles at the neurovascular unit: In vitro and in vivo studies to assess the blood-brain barrier permeability and function

Objective: Nanoparticle-based imaging and nanocarriers therapies have emerged as essential tools for many fields of modern medicine, in order to track the fate of cells and optimize drug delivery. Up to now, however, there are only few reports on the effect of nanocarriers of different types on oxygen delivery, even though this would be of great interest for the design of high impact therapies in several cardiovascular diseases (CVDs). In particular, Cyclodextrin Nanosponges (C-NS) can be envisioned as innovative tools to improve the delivery of oxygen in a controlled manner in CVDs. Methods: We tested oxygenated C-NS (OX-C-NS) at different concentrations (0.2, 2 and 20 μg/ml) for their capability to reduce cell mortality during hypoxia and reoxygenation (H/R) protocols. For comparative purpose, we also tested “blank materials” (C-NS filled with nitrogen gas without oxygen) and the effects of C-NS in Normoxia. To test the effectiveness of C-NS, we used H9c2, a cardiomyoblast cell line derived from rat heart, exposed to Normoxia (5% CO2 and 21% O2) or Hypoxia (5% CO2 and 95%N2) in a Hypoxic Chamber. The cellular mortality was measured with MTT assay. Results: In Normoxia, regardless of OX-C-NS formulation, the H9c2 cells displayed a tendency to an increased proliferation, which seemed somewhat correlated to the concentration of OX-C-NS used. The different concentration of OX-C-NS, applied before Hypoxia, induced a significant reduction of cell mortality compared to C-NS without oxygen. Also, the application of OX-C-NS at the beginning of reoxygenation induced a marked reduction of cell death. Conclusions: OX-C-NS may induce H9c2 cell proliferation in Normoxia and may protect H9c2 from H/R injury in vitro. The administration of oxygen in a controlled manner during or after an ischemic event may be an innovative approach for reduction of Ischemia/Reperfusion injury, with consequent reduction of chronic CVDs. Our preliminary results, and in particular the observation of a remarkable efficacy in reoxygenation, suggest an interesting potentiality for medical application of C-NS during the treatment of myocardial infarction. Further studies are required to ascertain the protective potential of C-NS on cardiac I/R injury under in vivo conditions.

Innovative Nanoparticles Enhance N-Palmitoylethanolamide Intraocular Delivery

Nanostructured lipid carriers (NLCs) loaded with palmitoylethanolamide (PEA) were formulated with the aim to enhance ocular bioavailability of PEA, particularly to the back of the eye. Technological characterization (e.g., size, charge) of NLC loaded with PEA formulation (PEA-NLC) was performed, and NLC morphology was characterized by electron microscopy. Ocular pharmacokinetic study, after topical administration of the formulation, was carried out in rabbit eye. Ultra-high performance liquid chromatography tandem mass spectrometry analysis was carried out to detect PEA levels in ocular tissues. Finally, the ocular tolerability of PEA-NLC formulation was assessed in rabbit eye. The novel formulation significantly increased PEA levels in ocular tissues compared to PEA suspension. Vitreous and retinal levels of PEA were significantly higher in the group treated with PEA-NLC formulation versus PEA suspension (PEA-NLC Cmax 5919 ± 541 pmol/g and 315 ± 70 pmol/g in vitreous and retina, respectively). The PEA-NLC formulation was characterized by high stability and robust ocular bioavailability. Therefore, this innovative formulation may be useful in clinical practice to manage retinal diseases.

Nanoparticles at the neurovascular unit: in vitro and in vivo studies to assess the blood-brain barrier permeability and function

Margarethe Geiger; Nandu Goswami; Michael Fischer; Markus Ritter; Marjan Slak Rupnik; Johann Wojta