Antioxidant Effect of the Castanea sativa Mill. Leaf Extract on Oxidative Stress Induced upon Human Spermatozoa

This study was aimed at evaluating in vitro the effects of a 75% v/v ethanolic extract of leaves of Castanea sativa Mill. (var. Bastarda Rossa, Mount Amiata, Tuscany, Italy) on ejaculated human sperm. Total polyphenols and total flavonoids contained in the extract were determined by a colorimetric assay and HPLC-DAD. The DPPH test and electrochemistry were utilized to study the antioxidant activity of the extract. Swim-up-selected sperm from 8 healthy men were treated with the C. sativa leaf extract at different dilutions (1: 100, 1: 200, and 1: 500), and sperm motility was assessed following the WHO guidelines. Swim-up-selected spermatozoa were incubated with 100 μM H2O2 to induce lipid peroxidation (LPO) and with H2O2 and the leaf extract (1: 100, 1: 200, and 1: 500) to test the antioxidant activity of the extract. The levels of LPO were determined by measuring malondialdehyde (MDA) concentrations. The treated samples were also analyzed by transmission electron microscopy (TEM) for ultrastructural evaluation. The chemical analysis showed that one-third ca. of the polyphenols in the C. sativa extract were made up of flavonoids, with hyperoside present in high concentration. A good antioxidant activity was demonstrated by both the DPPH test and electrochemical analysis. The C. sativa leaf extract did not decrease sperm motility at all tested dilutions. Treatment with H2O2 alone caused a significant increment in MDA levels (P=0.006993), while the treatment with H2O2 plus C. sativa extract diluted to 1: 100 and 1: 200 significantly reduced MDA levels (P=0.01476 and P=0.01571, respectively), with respect to H2O2 alone. TEM analysis confirmed the protective effect of the extract on damage induced by LPO, in particular that occurring at the plasma membrane level. The C. sativa leaf extract could be used in human and farm animal protocols for gamete handling, such as techniques of assisted reproduction and cryopreservation of semen, all conditions in which oxidative stress is exacerbated

Eudermic Properties and Chemical–Physical Characterization of Honeys of Different Botanical Origin

Honey is a natural product that, thanks to its composition, particularly the high sugar content, is highly appreciated as an energy nourishment. In addition to sugars, it contains many other substances (carbohydrates, free amino acids, enzymatic proteins, organic acids, polyphenols) from which the therapeutic properties of honey arise: hydrating and osmotic activity, antimicrobial action, and antioxidant and anti-inflammatory power. Objectives: The present work aims to deepen our knowledge/understanding of the activity of skin protection exerted by honey, as a synergic result of its multiple therapeutic effects. Moreover, this study wants to find possible correlations between biological properties and the chemical–physical traits of honey. Methods: To carry out this research, five varieties of citrus honey, one of acacia honey, one of chestnut honey, and one of multifloral honey were used. The honeys were first characterized by chemical–physical analysis and then were subjected to qualitative melissopalynological analysis. Tests were also carried out to evaluate both their antioxidant power and the effect on collagenase, an enzyme involved in the degradation of collagen present in the extracellular matrix and, therefore, in the processes of skin aging. Finally, honey samples were then used in in vitro experiments to assess their action in stimulating cell viability and proliferation on human keratinocytes. Results: Chemical–physical analysis demonstrated a good water content (about 17%), an important sugar content (with the monosaccharides glucose and fructose being the most represented in all the honey samples), various amino acids (with proline remarkably being the highest in all honeys), and a high concentration of polyphenols and total flavonoids (the maximum in chestnut honey, 762 mg/kg and 514 mg/kg, respectively). Conclusions The results obtained in this work confirm the ethnopharmacological use of honey in wound care, bring new scientific knowledge on the use of honey in dermatology, and highlight two fields of excellence, particularly incitrus and chestnut honey

Gradients of Variation in the At-Vessel Mortality Rate between Twelve Species of Sharks and Skates Sampled through a Fishery-Independent Trawl Survey in the Asinara Gulf (NW Mediterranean Sea)

Elasmobranchs are priority species for conservation due to their rapid decline determined by the unbalanced struggle between a fragile bio-ecology and strong anthropogenic impacts, such as bycatch from professional fishing. In this context, measuring species resistance to catch of poorly selective gear is of paramount importance. During June-October 2022, five experimental fishing campaigns were carried out in the Asinara Gulf (northern Sardinia) through 35 geographically and bathymetrically representative hauls of an area between 30 and 600 m in depth. Skates prevailed over sharks in the number of species, with seven and five species, respectively. We first evaluated the status of each individual with respect to stress due to the trawl's catch using a three-graded scale. We also recorded individual biometrics (total and disk length, weight and sex, and maturity for males) on board by implementing the best practices in manipulating individuals for physiological recovery and release at sea. After capture, skates resulted in generally better conditions than sharks, although deepwater species of both groups exhibited a worse state than coastal species. The estimated vitality rates also depended on the size of the individuals. This work provides standardized data on the intermingled effect of size, species type, and inhabited depth on the resistance response of some elasmobranch species against capture by trawl fishery activities

A Novel Perilla frutescens (L.) Britton Cell-Derived Phytocomplex Regulates Keratinocytes Inflammatory Cascade and Barrier Function and Preserves Vaginal Mucosal Integrity In Vivo

: In the last years, the medicinal plant Perilla frutescens (L.) Britton has gained scientific interest because leaf extracts, due to the presence of rosmarinic acid and other polyphenols, have shown anti-allergic and skin protective potential in pre-clinical studies. Nevertheless, the lack of standardized extracts has limited clinical applications to date. In this work, for the first time, a standardized phytocomplex of P. frutescens, enriched in rosmarinic acid and total polyphenols, was produced through innovative in vitro cell culture biotechnology and tested. The activity of perilla was evaluated in an in vitro inflammatory model of human keratinocytes (HaCaT) by monitoring tight junctions, filaggrin, and loricrin protein levels, the release of pro-inflammatory cytokines and JNK MAPK signaling. In a practical health care application, the perilla biotechnological phytocomplex was tested in a multilayer model of vaginal mucosa, and then, in a preliminary clinical observation to explore its capacity to preserve vaginal mucosal integrity in women in peri-menopause. In keratinocytes cells, perilla phytocomplex demonstrated to exert a marked activity in epidermis barrier maintenance and anti-inflammatory effects, preserving tight junction expression and downregulating cytokines release through targeting JNK activation. Furthermore, perilla showed positive effects in retaining vaginal mucosal integrity in the reconstructed vaginal mucosa model and in vivo tests. Overall, our data suggest that the biotechnological P. frutescens phytocomplex could represent an innovative ingredient for dermatological applications

Study of the effects of Lemna minor extracts on human immune cell populations

OBJECTIVE: Lemna minor is a plant with a huge repertoire of secondary metabolites. The literature indicates that extracts of Lemna minor have antioxidant, antiradical, immunomodulatory and anti-inflammatory properties. The objective of the present study was to find a suitable technique to extract active compounds from this plant and verify whether these extracts have immunomodulatory activity. MATERIALS AND METHODS: We grew L. minor on a standard medium with Gamborg B5 and vitamins. We extracted compounds from the plant by maceration and decoction. The phytochemical profile of the extracts was characterized by chromatography, spectrophotometry, and spectroscopy. The extracts were tested on cultures of mononuclear cells from four human subjects. These cells were pulsed with carboxyfluorescein succinimidyl ester, grown in triplicate in standard culture medium without (control) and with increasing concentrations of Lemna extracts. Flow cytometry was used to evaluate cell death and proliferation of the total mononuclear cell population and of CD4+, CD8+, B cell and monocyte populations. RESULTS: The Lemna extracts were not cytotoxic and did not cause cell necrosis or apoptosis in immune cells. At low concentrations, they induced very limited proliferation of CD4+ cells within 48 hours. At high concentrations, they induced proliferation of CD8+ cells and B lymphocytes within 48 hours. CONCLUSIONS: Unfortunately, we failed to confirm any immunomodulatory activity of Lemna extracts. Growth and death rates of human immune cells were not significantly affected by adding Lemna extracts to the culture medium

Quality of Reporting on the Vegetative State in Italian Newspapers. The Case of Eluana Englaro

Background: Media coverage of the vegetative state (VS) includes refutations of the VS diagnosis and describes behaviors inconsistent with VS. We used a quality score to assess the reporting in articles describing the medical characteristics of VS in Italian newspapers. Methodology/Principal Findings: Our search covered a 7-month period from July 1, 2008, to February 28, 2009, using the online searchable databases of four major Italian newspapers: Corriere della Sera, La Repubblica, La Stampa, and Avvenire. Medical reporting was judged as complete if three core VS characteristics were described: patient unawareness of self and the environment, preserved wakefulness (eyes open), and spontaneous respiration (artificial ventilator not needed). We retrieved 2,099 articles, and 967 were dedicated to VS. Of these, 853 (88.2%) were non-medical and mainly focused on describing the political, legal, and ethical aspects of VS. Of the 114 (11.8%) medical articles, 53 (5.5%) discussed other medical problems such as death by dehydration, artificial nutrition, neuroimaging, brain death, or uterine hemorrhage, and 61 (6.3%) described VS. Of these 61, only 18 (1.9%) reported all three CORE characteristics and were judged complete. We found no differences among the four investigated newspapers (Fisher’s exact = 0.798), and incomplete articles were equally distributed between journalistic pieces and expert opinions (x 2 = 1.8854, P = 0.170). Incorrect descriptions of VS were significantly more common among incomplete articles (13 of 43 vs. 1 of 18; Fisher’s exact P = 0.047)

Olea europaea leaf extract: antioxidant properties and supplement in human sperm cryopreservation

Background: Semen handling and cryopreservation increase reactive oxygen species (ROS) production, exposing spermatozoa to oxidative stress (OS) that can be minimised by antioxidant supplementation. We studied if Olea europaea leaf extract (OE) could have antioxidant and protective activity during sperm manipulation.Methods: The extract was characterized by high-performance liquid chromatography with diode array detection (HPLC-DAD) and antioxidant activity was determined by 2,2-diphenyl-1-picrylhydrazyl (DPPH) and oxygen radical absorbance capacity (ORAC) assays. Then, spermatozoa of 35 normozoospermic donors were treated with OE. First, swim-up selected spermatozoa were incubated with OE (1:100-1:400) and sperm motility and DNA integrity (acridine orange test) were assessed. Then, swim-up selected sperm were treated with 100 mu M H2O2 to induce OS with and without OE; motility, DNA integrity and F-2-Isoprostanes (F-2-IsoPs quantified by gas chromatography/negative ion chemical ionization tandem mass spectrometry analysis), an OS marker, were assessed. Finally, sperm were frozen with and without OE. Beside the previous endpoints, acrosome shape was evaluated by Tetramethylrhodamine (TRITC)-conjugated Pisum sativum agglutinin (PSA).Results: OE was enriched in polyphenols (0.34%) and triterpenes (0.72%) and the synergistic action of phytocomplex components was responsible for antioxidant activity. Since OE was not toxic for spermatozoa, 1:100 dilution was used for the other experiments. OE protected motility, DNA integrity and reduced F-2-IsoPs (p < 0.001) in in vitro experiment with OS induction. OE treated frozen-thawed spermatozoa showed increased motility, DNA integrity, reduced F-2-IsoP concentration (p < 0.001) and normal acrosomes versus non-supplemented samples.Conclusions: OE is characterized by high concentration of polyphenols and exhibits protective properties against oxidative damage induced by H2O2 in human ejaculated sperm. After cryopreservation, the samples supplemented with the extract showed increased sperm quality. OE could represent a supplement of culture media during semen handling where OS is exacerbated