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

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)

Safety and Efficacy of Red Yeast Rice Phytocomplex and Lovastatin: a Comparative Analysis

In the last 20 years, the demand for red yeast rice-based food supplements has significantly increased as consumers have been looked for natural alternatives to manage blood cholesterol levels without the adverse effects associated with synthetic statins. Actually, monacolin K, derived from red yeast rice (RYR) and chemically identical to lovastatin, inhibits 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase, offering a natural alternative to synthetic drugs. In 2018, despite the health claim approved in 2011, the European Food Safety Authority (EFSA) raised concerns about the safety of monacolin K consumption, especially for vulnerable populations, and highlighted the lack of knowledge on RYR phytocomplex. Consequently, in 2021, the European Commission established a new maximum intake limit < 2.99 mg/day for monacolins. This study aimed to investigate the safety and efficacy of RYR phytocomplex compared to the sole monacolin K (=lovastatin) by analyzing eight different RYR samples with different monacolin K and secondary monacolins content. Efficacy was evaluated through a validated cell-free enzymatic assay, while an integrated in vitro simulated digestion and in silico ADME prediction were employed to compare the pharmacokinetics of different samples and lovastatin. The safety of RYR was assessed by monitoring cytotoxicity in intestinal, hepatic, kidney, and skeletal muscle cells using cell viability assays. Furthermore, muscle damage-related targets and myokines were measured by qRT-PCR in myoblasts exposed to prolonged non-toxic stimuli. Results demonstrated that RYR samples have a large chemical variability not only related to monacolin K content, in lactone and hydroxy-acid form, but also as regards pigments, secondary monacolins, polyphenols, and triterpenes. The enzymatic test revealed that all samples were more effective in inhibiting HMG-CoA activity than lovastatin at equivalent monacolin K content. Although secondary monacolins had a weaker effect than monacolin K, a synergistic effect was observed within the phytocomplex. Additionally, the phytocomplex facilitated higher bioaccessibility of monacolin K in RYR compared to lovastatin. Regarding cell viability, lovastatin and RYR samples exhibited no toxic effects up to 150 g/mL on intestinal, hepatic, and kidney cells; however, variable effects were observed on skeletal muscle cells at medium-high concentrations. Nevertheless, at physiological non-toxic concentrations, RYR samples did not impact the expression levels of myokine IL-6, autophagy factors, and catabolic markers of protein balance, unlike synthetic statin. These results suggest that RYR and lovastatin share similar biological activities, but the phytocomplex's contribution from secondary monacolins, pigments, polyphenols, and triterpenes ensure a superior safety profile for this botanical food supplement. However, it is important to note that the composition of RYR phytocomplex can vary significantly between different products on the market. This variability highlights the need for standardization and quality control measures to ensure the safety and efficacy of this botanical food supplement. In conclusion, our study provides evidence supporting the enhanced efficacy, bioaccessibility, and safety profile of RYR phytocomplex compared to lovastatin demonstrating its potential as a natural and effective option for hypercholesterolemia management

In vitro neuraminidase inhibitory effect and activity against influenza virus A H1N1 of herbal drugs used for common cold

Influenza viruses are causative agents of common respiratory infections, often underestimated and badly managed. Monomolecular anti-inflammatory and anti-pyretic drugs are extensively used in infection related symptoms, but they do not trigger virus replication. Also herbal drugs are traditionally used [1], although specific anti influenza activities are rarely recorded and little investigated. By means of validated in vitro tests, in this study we investigated the effect of ten herbal extracts and poplar propolis commonly used for respiratory infections. We tested their effectiveness against an influenza A H1N1 strain in pre- and post cell infection and neuraminidase activity inhibition. Camellia sinensis Kuntze leaves dried extract 60% catechins (CSE), Pelargonium sidoides DC. roots standardized d.e. (PS) and Cistus creticus L. subsp. eriocephalus (Viv.) Greuter & Burdet (formerly Cistus incanus L.) aerial parts commercial d.e. 20% polyphenols (CIS) showed a strong anti-neuraminidase effect (IC50<25 μg/ml). Only these extracts were efficacious in in vitro inhibiting influenza virus entry and (in a weaker manner) virus replication in MDCK cells; toxic/effective concentration ratio was >4 only for CIS. Chemical analyses showed that galloil derivatives and flavan-3-ols oligomers mainly occur in CIS and, similarly to PS and CSE, these constituents are likely to contribute more to the antiviral efficacy [2], [3]. Our findings suggest that C. creticus subsp. eriocephalus is worth to be better investigated for its anti-influenza efficacy and ongoing researches are aiming to investigate molecular mechanisms of CIS constituents on viral targets such as the influenza virus glycoproteins, emoagglutinin and neuraminidase