Mediterranean Shrub Species as a Source of Biomolecules against Neurodegenerative Diseases.

Neurodegenerative diseases are associated with oxidative stress, due to an imbalance in the oxidation-reduction reactions at the cellular level. Various treatments are available to treat these diseases, although they often do not cure them and have many adverse effects. Therefore, it is necessary to find complementary and/or alternative drugs that replace current treatments with fewer side effects. It has been demonstrated that natural products derived from plants, specifically phenolic compounds, have a great capacity to suppress oxidative stress and neutralize free radicals thus, they may be used as alternative alternative pharmacological treatments for pathological conditions associated with an increase in oxidative stress. The plant species that dominate the Mediterranean ecosystems are characterized by having a wide variety of phenolic compound content. Therefore, these species might be important sources of neuroprotective biomolecules. To evaluate this potential, 24 typical plant species of the Mediterranean ecosystems were selected, identifying the most important compounds present in them. This set of plant species provides a total of 403 different compounds. Of these compounds, 35.7% are phenolic acids and 55.6% are flavonoids. The most relevant of these compounds are gallic, vanillic, caffeic, chlorogenic, p-coumaric, and ferulic acids, apigenin, kaempferol, myricitrin, quercetin, isoquercetin, quercetrin, rutin, catechin and epicatechin, which are widely distributed among the analyzed plant species (in over 10 species) and which have been involved in the literature in the prevention of different neurodegenerative pathologies. It is also important to mention that three of these plant species, Pistacea lentiscus, Lavandula stoechas and Thymus vulgaris, have most of the described compounds with protective properties against neurodegenerative diseases. The present work shows that the plant species that dominate the studied geographic area can provide an important source of phenolic compounds for the pharmacological and biotechnological industry to prepare extracts or isolated compounds for therapy against neurodegenerative diseases.

Influences of Yogurt with Functional Ingredients from Various Sources That Help Treat Leaky Gut on Intestinal Barrier Dysfunction in Caco-2 Cells.

The impact of yogurts made with starter culture bacteria (L. bulgaricus and S. thermophilus) and supplemented with ingredients (maitake mushrooms, quercetin, L-glutamine, slippery elm bark, licorice root, N-acetyl-D-glucosamine, zinc orotate, and marshmallow root) that can help treat leaky gut were investigated using the Caco-2 cell monolayer as a measure of intestinal barrier dysfunction. Milk from the same source was equally dispersed into nine pails, and the eight ingredients were randomly allocated to the eight pails. The control had no ingredients. The Caco-2 cells were treated with isoflavone genistein (negative control) and growth media (positive control). Inflammation was stimulated using an inflammatory cocktail of cytokines (interferon-γ, tumor necrosis factor-α, and interleukin-1ß) and lipopolysaccharide. The yogurt without ingredients (control yogurt) was compared to the yogurt treatments (yogurts with ingredients) that help treat leaky gut. Transepithelial electrical resistance (TEER) and paracellular permeability were measured to evaluate the integrity of the Caco-2 monolayer. Transmission electron microscopy (TEM), immunofluorescence microscopy (IM), and real-time quantitative polymerase chain reaction (RTQPCR) were applied to measure the integrity of tight junction proteins. The yogurts were subjected to gastric and intestinal digestion, and TEER was recorded. Ferrous ion chelating activity, ferric reducing potential, and DPPH radical scavenging were also examined to determine the yogurts' antioxidant capacity. Yogurt with quercetin and marshmallow root improved the antioxidant activity and TEER and had the lowest permeability in fluorescein isothiocyanate (FITC)-dextran and Lucifer yellow flux among the yogurt samples. TEM, IM, and RTQPCR revealed that yogurt enhanced tight junction proteins' localization and gene expression. Intestinal digestion of the yogurt negatively impacted inflammation-induced Caco-2 barrier dysfunction, while yogurt with quercetin, marshmallow root, maitake mushroom, and licorice root had the highest TEER values compared to the control yogurt. Yogurt fortification with quercetin, marshmallow root, maitake mushroom, and licorice root may improve functionality when dealing with intestinal barrier dysfunction.

Effect of Microwave and Ultrasound-Assisted Extraction on the Phytochemical and In Vitro Biological Properties of Willow (Salix alba) Bark Aqueous and Ethanolic Extracts.

White willow (Salix alba) is a medicinal plant used in folk medicine. In this study, aqueous and ethanolic willow bark extracts were obtained via ultrasonic-assisted extraction (UAE) and microwave-assisted extraction (MAE), and analyzed regarding their phytochemical (total phenolics, phenolic acids, flavonoids, and tannins) content and in vitro biological properties (antibacterial and antifungal activity, acetylcholinesterase AChE inhibitory activity and anti-inflammatory effects). The highest phenolic, tannin, and flavonoid contents were found for willow bark extracts obtained via microwave-assisted extraction using ethanol as a solvent (SA-ME). The polyphenol load of all MAE and UAE extracts was higher when conventional solid-liquid extraction was applied (ρ < 0.05). The antioxidant capacities were stronger for microwave-assisted ethanolic extracts, with the lowest IC50 values of 12 µg/mL for DPPH• and a value of 16 µg/mL for ABTS•+, whereas the conventional extraction had the highest IC50 values (22 µg/mL and 28 µg/mL, respectively). Willow bark extract showed antibacterial activity against Gram-positive bacteria S. aureus and P. aeruginosa. AChE inhibitory activity was dependent on the extraction method and solvent used, and the highest inhibition among samples was observed for SA-ME. Taken altogether, our findings suggest that willow (Salix alba) bark extract obtained via ethanolic microwave-assisted extraction is a phytochemical-rich resource with in vitro, anti-inflammatory, and AchE inhibitory properties and, therefore, potential multiple medicinal end-uses.