ESTRATTI DI MACROALGHE ERICARIA CRINITA, ERICARIA BRACHYCARPA E ASPARAGOPSIS TAXIFORMIS: DAGLI EFFETTI SUL RICCIO DI MARE ARBACIA LIXULA (LINNEO, 1758) ALLE POTENZIALI APPLICAZIONI

Lo studio delle macroalghe è essenziale per la comprensione degli ecosistemi marini e per lo sfruttamento delle risorse economiche che esse rappresentano come fonte di biomolecole ad alto valore biologico. Questo studio ha comportato l'estrazione e la caratterizzazione chimica dei metaboliti secondari presenti nelle specie di macroalghe Ericaria crinita, Ericaria brachycarpa e Asparagopsis taxiformis e la valutazione delle loro proprietà biologiche. La caratterizzazione dei metaboliti secondari ha permesso l'identificazione dei composti: prevalentemente ossilipine negli estratti di E. brachycarpa, meroterpenoidi in E. crinita e composti bromurati in A. taxiformis. In particolare, le analisi hanno permesso di determinare un elevato contenuto fenolico negli estratti della specie E. crinita. Successivamente, gli estratti sono stati utilizzati per valutare l’attività antimicrobica contro alcuni ceppi batterici di Listeria monocytogenes e Staphylococcus aureus e l’attività citotossica verso eritrociti di mammifero, cellule di neuroblastoma ed epatocarcinoma. In particolare, per quanto riguarda l'attività antibatterica, gli estratti di E. brachycarpa e A. taxiformis agiscono ad ampio spettro nei confronti dei ceppi di Listeria monocytogenes e Staphylococcus aureus. Questi estratti hanno mostrato una bassa attività emolitica nei confronti degli eritrociti di mammifero (< 5%) a concentrazioni ≤ 40 μg/ml.Successivamente gli estratti di E. crinita ed E. brachycarpa sono stati testati per valutare l’attività antitumorale nei confronti delle linee cellulari tumorali HepG2 e SH-SY5Y. I risultati ottenuti hanno dimostrato che alle concentrazioni testate gli estratti di E. crinita ed E. brachycarpa hanno esercitato attività antitumorale verso la linea tumorali HepG2, mentre il dosaggio antitumorale degli estratti di E. brachycarpa ha mostrato l’importante attività citotossica verso la linea tumorale SH-SY5Y.Infine, sulla base dei risultati precedentemente ottenuti, è stata valutata l'attività miorilassante (ex vivo) degli estratti della macroalga A. taxiformis sulla muscolatura liscia del colon di Rattus norvegicus. L'estratto di A. taxiformis è stato in grado di regolare la motilità intestinale di R. norvegicus mostrando attività antispasmodica.Sulla base dei risultati ottenuti si è deciso di valutare gli effetti di questi estratti utilizzando l'organismo modello A. lixula, uno dei responsabili della degradazione delle Cistoseire in ambiente marino. La conta cellulare, i livelli di ROS, NO, GSH, la concentrazione di proteine totali e l'attività enzimatica di esterasi e fosfatasi alcalina sono stati utilizzati come parametri biologici del fluido celomatico negli animali di controllo e trattati. I risultati hanno mostrato importanti cambiamenti dei livelli di ROS, NO, GSH e delle attività enzimatiche dopo 3 ore di trattamento in vivo. L'attività citotossica degli estratti algali sullo sviluppo embrionale di A. lixula è stata analizzata solo dopo aver studiato la loro attività antitumorale nei confronti delle linee cellulari tumorali HepG2 e SH-SY5Y, testando 4 concentrazioni (1,4,20,40 μg/ml) a 24, 48 e 72 ore dopo la fecondazione di ricci di mare. I risultati ottenuti hanno mostrato come l'estratto di E. brachycarpa, alla concentrazione di 40 μg/ml, provoca una mortalità (apoptosi) del 100% degli embrioni di A. lixula. Infatti, gli embrioni esposti alla concentrazione più alta di E. brachycarpa (40 μg/ml) degenerano, mentre a concentrazioni più basse (1-4 μg/ml) l'estratto manifesta attività, ritardando lo sviluppo embrionale dei ricci e mostrando variazioni significative morfologiche nelle diverse fasi di sviluppo degli embrioni.Sulla base della letteratura e dei risultati precedentemente ottenuti su A. lixula, è stato condotto uno studio sull'interazione trofica tra il riccio di mare Arbacia lixula e due specie di macroalghe, E. crinita e Caulerpa taxifolia var. disticophilla. È emerso che A. lixula è in grado di pascolare entrambe le specie di macroalghe, mostrando una preferenza per quella non autoctona. Questo risultato ha fornito una base importante per comprendere il possibile ruolo di A. lixula nel contenere la diffusione delle macroalghe aliene.In conclusione, i risultati ottenuti dimostrano come i composti presenti nelle specie di macroalghe testate possiedano proprietà antiossidanti che potrebbero essere utilizzate per l’industria alimentare e zootecnica, potenzialmente in grado di migliorare il livello di antiossidanti e consentire una maggiore conservabilità dei prodotti alimentari. Inoltre, le potenziali applicazioni, per quanto riguarda l'industria farmaceutica, sono di notevole interesse, poiché questi composti permetterebbero il potenziale sviluppo di farmaci con azione terapeutica antinfiammatoria, antitumorale, antispasmodica intestinale e antibiotica. Infine, i risultati ottenuti hanno mostrato per la prima volta la presenza di ossilipine nell'estratto di E. brachycarpa e la sua possibile influenza sullo sviluppo embrionale di A. lixula, suggerendo un potenziale meccanismo di difesa che potrebbe essere messo in atto dalla Cystoseira contro i suoi erbivori.The study of macroalgae is essential for the understanding of marine ecosystems, and for the exploitation of the economic resources that they represent as a source of biomolecules with high biological value. This study involved the extraction and chemical characterization of secondary metabolites present in the macroalgae species Ericaria crinita, Ericaria brachycarpa and Asparagopsis taxiformis and the evaluation of their biological properties. The characterization of secondary metabolites allowed the identification of compounds: predominantly oxylipin in E. brachycarpa extracts, meroterpenoids in E. crinita and brominated compounds in A. taxiformis. In particular, the analyses allowed us to determine a high phenolic content in the extracts in the E. crinita species. Subsequently, the extracts were used to evaluate the antimicrobial activity against some bacterial strains of Listeria monocytogenes and Staphylococcus aureus and cytotoxic activity vs mammalian eritrocytes, neuroblastoma and hepatocarcinoma cells. Regarding antibacterial activity, the extracts of E. brachycarpa and A. taxiformis they act across a broad spectrum towards Listeria monocytogenes and Staphylococcus aureus strains. These extracts showed a low haemolytic activity towards mammalian erythrocytes (< 5%) at concentrations ≤ 40 μg/ml. Subsequently, E. crinita and E. brachycarpa extracts were tested to evaluate the antitumor activity against HepG2 and SH-SY5Y tumor cell lines. The results obtained demonstrated that at the tested concentrations the E. crinita and E. brachycarpa extracts exerted anti-tumor activity towards HepG2 tumor cell line, while the anti-tumor dosage of E. brachycarpa extracts showed the important cytotoxic activity towards tumor cell line SH-SY5.Finally, based on the results previously obtained, the muscle relaxant activity (ex vivo) of the extracts of the macroalgae A. taxiformis on the smooth muscles of the colon of Rattus norvegicus was evaluated. A. taxiformis extract was able to regulate the intestinal motility of R. norvegicus showing antispasmodic activity.Based on the results obtained, it was decided to evaluate the effects of these extracts using the model organism Arbacia lixula, one of those responsible for the degradation of Cystoseire in the marine environment. Cell counts, levels of ROS, NO, GSH, total protein concentration and enzymatic activity of esterase and alkaline phosphatase were used as biological parameters of the haemolymph in control and treated animals. The results showed important changes in the enzymatic activities after 3 hours of in vivo treatment. The cytotoxic activity of algal extracts on the embryonic development of A. lixula was analyzed only after studying their anti-tumor activity against the tumor cell lines HepG2 and SH-SY5Y, testing 4 concentrations (1,4,20,40 μg/ml) at 24, 48 and 72 hours after fertilization of sea urchins. The results obtained showed how the E. brachycarpa extract, at a concentration of 40 μg/ml, causes 100% mortality (apoptosis) of A. lixula embryos. In fact, the embryos exposed to the highest concentration of E. brachycarpa (40 μg/ml) degenerate, while at lower concentrations (1-4 μg/ml) the extract shows activity, delaying the embryonic development of the urchins and showing significant characteristics morphological variations in the different stages of development of the embryos. Based on the literature and results previously obtained on A. lixula, a study was conducted on the trophic interaction between the sea urchin Arbacia lixula and two species of macroalgae, E. crinita and Caulerpa taxifolia var. disticophilla. It emerged that A. lixula is capable of grazing both species of macroalgae, showing a preference for the non-native one. This result provided an important basis for understanding the possible role of A. lixula in containing the spread of alien macroalgae. In conclusion, the results obtained demonstrate how the compounds present in the tested macroalgae species possess antioxidant properties that could be used for the food and livestock industry, potentially capable of improving the level of antioxidants and allowing a greater shelf life of food products. Furthermore, the potential applications, regarding the pharmaceutical industry, are considerable interest, since these compounds would allow the potential development of drugs with anti-inflammatory, anti-tumor, intestinal antispasmodic, and antibiotic therapeutic action. Finally, the results obtained for the first time showed the presence of oxylipin in the E. brachycarpa extract and his possible influence on the embryonic development of A. lixula, suggesting a potential defence mechanism that could be implemented by the Cystoseira against its herbivores

Environmental DNA: The First Snapshot of the Vertebrate Biodiversity in Three Sicilian Lakes

Freshwater ecosystems play a key role in global diversity and are subject to a series of anthropic impacts, often leading to biodiversity loss. The organisms inhabiting these sites continuously release DNA into the environment through cells, excrement, gametes and/or decomposing matter; thus, evaluation of this eDNA could revolutionize the monitoring of biodiversity. In this study, environmental DNA metabarcoding was used for the first time in three Sicilian lakes: Lake Poma, Piana degli Albanesi Lake and Lake Scanzano. Results obtained provide the first snapshot of vertebrate biodiversity in these three lakes, where little is known, to provide valuable information useful for creating a baseline of knowledge regarding the biodiversity in these three lakes. Another important result was the detection of marine species, most likely due to some kind of anthropogenic contamination. Environmental DNA is a useful tool to evaluate both biodiversity and the ecological status of the environment; it has the potential to complement traditional methods, and the use of both approaches may offer a more comprehensive understanding of the ecosystem

Chemical Compositions and Antioxidant Activities of Essential Oils, and Their Combinations, Obtained from Flavedo By-Product of Seven Cultivars of Sicilian Citrus aurantium L.

In this work, seven Citrus aurantium essential oils (EOs) derived from flavedo of cultivars ‘Canaliculata’, ‘Consolei’, ‘Crispifolia’, ‘Fasciata’, ‘Foetifera’, ‘Listata’, and ‘Bizzaria’ were investigated. EOs were also combined in 1:1 (v/v) ratio to identify possible synergism or antagonism of actions. GC-MS analysis was done to investigate Eos’ phytochemical profiles. The antioxidant activity was studied by using a multi-target approach based on FRAP, DPPH, ABTS, and β-carotene bleaching tests. A great difference was observed in EOs’ phytochemical profiles. d-limonene (33.35–89.17%) was the main monoterpene hydrocarbon, and α-Pinene, β-myrcene, and β-linalool were identified in almost all samples. Among EOs, only C3 showed high quantitative and qualitative variability in its chemical composition. The chemical diversity of EOs was also demonstrated by PCA and HCA statistical analysis. Samples C2, C4, C5, C6, and C7 were statistically similar to each other, while C1 and C3 were characterized as having a different amount of other compounds and oxygenated monoterpenes, respectively, with respect to the other EOs mentioned. The global antioxidant score (GAS) revealed that among the tested EOs, C. aurantium ‘Fasciata’ EO had the highest antioxidant potential, with a GAS value of −0.47, whereas among combinations, the EO obtained by mixing ‘Canaliculata’ + ‘Bizzaria’ was the most active. Comparison by theoretical and real data on inhibitory concentration (IC50) and FRAP values did not reveal any significant effect of synergism or antagonism of actions to be valid in all biological applied tests. These findings, considered together, represent an important starting point to understand which compounds are responsible for the activities and their future possible industrial application

The Effects of Red-Grape Seed and Pomace-Flour Dietary Supplementation on Broiler Chickens

Wine by-products, generated in large quantities and wasted with serious environmental and economic consequences, are an exceptional alternative to conventional and non-sustainable food sources. The aim of the study was to understand if these by-products could be reused in feed for broiler chickens, improving certain blood parameters important in immune responses, by testing different concentrations and administration times. The effects of the grape-seed and pomace flour of red-wine waste were evaluated. Three different diets were tested containing pomace (0%, 3%, and 6%), grape seeds (0%, 3%, and 6%), and a mix of pomace and grape seeds (0 and 3%) for 7, 21, and 42 days. Esterase, alkaline phosphatase, and peroxidase activity, along with glucose, reactive oxygen species, and glutathione levels, were evaluated in blood samples. The results showed significant changes in the parameters analyzed, especially after 21 and 42 days of administration. All diets administered showed significant effects based on exposure time and by-product concentration used. The greatest effects seem to have come from grape-seed flour. All the by-products of wine production analyzed in this study showed important antioxidant activities and can be included in the food context of farm animals in different growth phases at different concentrations

Shedding light on typical species : implications for habitat monitoring

Habitat monitoring in Europe is regulated by Article 17 of the Habitats Directive, which suggests the use of typical species to assess habitat conservation status. Yet, the Directive uses the term “typical” species but does not provide a definition, either for its use in reporting or for its use in impact assessments. To address the issue, an online workshop was organized by the Italian Society for Vegetation Science (SISV) to shed light on the diversity of perspectives regarding the different concepts of typical species, and to discuss the possible implications for habitat monitoring. To this aim, we inquired 73 people with a very different degree of expertise in the field of vegetation science by means of a tailored survey composed of six questions. We analysed the data using Pearson's Chi-squared test to verify that the answers diverged from a random distribution and checked the effect of the degree of experience of the surveyees on the results. We found that most of the surveyees agreed on the use of the phytosociological method for habitat monitoring and of the diagnostic and characteristic species to evaluate the structural and functional conservation status of habitats. With this contribution, we shed light on the meaning of “typical” species in the context of habitat monitoring

Effect of angiotensin-converting enzyme inhibitor and angiotensin receptor blocker initiation on organ support-free days in patients hospitalized with COVID-19

IMPORTANCE Overactivation of the renin-angiotensin system (RAS) may contribute to poor clinical outcomes in patients with COVID-19. Objective To determine whether angiotensin-converting enzyme (ACE) inhibitor or angiotensin receptor blocker (ARB) initiation improves outcomes in patients hospitalized for COVID-19. DESIGN, SETTING, AND PARTICIPANTS In an ongoing, adaptive platform randomized clinical trial, 721 critically ill and 58 non–critically ill hospitalized adults were randomized to receive an RAS inhibitor or control between March 16, 2021, and February 25, 2022, at 69 sites in 7 countries (final follow-up on June 1, 2022). INTERVENTIONS Patients were randomized to receive open-label initiation of an ACE inhibitor (n = 257), ARB (n = 248), ARB in combination with DMX-200 (a chemokine receptor-2 inhibitor; n = 10), or no RAS inhibitor (control; n = 264) for up to 10 days. MAIN OUTCOMES AND MEASURES The primary outcome was organ support–free days, a composite of hospital survival and days alive without cardiovascular or respiratory organ support through 21 days. The primary analysis was a bayesian cumulative logistic model. Odds ratios (ORs) greater than 1 represent improved outcomes. RESULTS On February 25, 2022, enrollment was discontinued due to safety concerns. Among 679 critically ill patients with available primary outcome data, the median age was 56 years and 239 participants (35.2%) were women. Median (IQR) organ support–free days among critically ill patients was 10 (–1 to 16) in the ACE inhibitor group (n = 231), 8 (–1 to 17) in the ARB group (n = 217), and 12 (0 to 17) in the control group (n = 231) (median adjusted odds ratios of 0.77 [95% bayesian credible interval, 0.58-1.06] for improvement for ACE inhibitor and 0.76 [95% credible interval, 0.56-1.05] for ARB compared with control). The posterior probabilities that ACE inhibitors and ARBs worsened organ support–free days compared with control were 94.9% and 95.4%, respectively. Hospital survival occurred in 166 of 231 critically ill participants (71.9%) in the ACE inhibitor group, 152 of 217 (70.0%) in the ARB group, and 182 of 231 (78.8%) in the control group (posterior probabilities that ACE inhibitor and ARB worsened hospital survival compared with control were 95.3% and 98.1%, respectively). CONCLUSIONS AND RELEVANCE In this trial, among critically ill adults with COVID-19, initiation of an ACE inhibitor or ARB did not improve, and likely worsened, clinical outcomes. TRIAL REGISTRATION ClinicalTrials.gov Identifier: NCT0273570

LA SCALA DEL BAGNO DI SIRACUSA NELLA CHIESA DI SAN FILIPPO APOSTOLO, DALLA RISCOPERTA AL RILIEVO DIGITALE

This article analyzes the helicoidal underground staircase of the Bath of Syracuse, accessible from the church of St. Philip the Apostol, carried out through the study of its eighteenth-century "rediscovery" and the accurate survey of the artifact, made with digital tools. The article interpretates the sources that demonstrate the archaeological attention addressed to this staircase, hypothesizing its direct knowledge already before the 18th century, that could testify its role as a model for other constructions

The Portuguese Man-of-War Has Always Entered the Mediterranean Sea—Strandings, Sightings, and Museum Collections

A search of records from different kinds of sources namely, scientific and grey literature, social media, and zoological museum collections, has been carried out to review the incidence of Physalia physalis (Linnaeus, 1758), the Portuguese man-of-war, in the Mediterranean Sea. The temporal frame of the records, considered valid if documented with images or collected specimens, ranged from the second half of the eighteenth century to the year 2021. Thanks to colonies preserved in some Italian historical museum collections, originating from the western basin, it was possible to date the putative first documented record of P. physalis of the Mediterranean Sea in 1850. The dataset shows some massive strandings that occurred in localities of the Strait of Gibraltar and Alboran Sea, the area of the entrance, from the Atlantic toward the Mediterranean waters, and the starting point from where the species spread toward the western and central basin. Physalia physalis does not reach the eastern area of the Mediterranean Sea. As the records of this species from the Italian maritime regions were abundant in the summertime and considering the danger related to contact with humans, they were subdivided into three categories of risk according to the months of occurrence. These categories were created to assign a level of danger for swimmers. The increasing sightings of such a poisonous cnidarian in coastal waters can represent a risk to human health, and a threat to all those activities linked to the marine tourism sector. The overview given goes beyond scientific purposes and aims to reach society and public administrators. The involvement of citizens and touristic structures for the early detection of P. physalis can play a key role in preventing encounters with the species, allowing marine tourist facilities to operate within a range of reasonable security

The Portuguese Man-of-War Has Always Entered the Mediterranean Sea—Strandings, Sightings, and Museum Collections

A search of records from different kinds of sources namely, scientific and grey literature, social media, and zoological museum collections, has been carried out to review the incidence of Physalia physalis (Linnaeus, 1758), the Portuguese man-of-war, in the Mediterranean Sea. The temporal frame of the records, considered valid if documented with images or collected specimens, ranged from the second half of the eighteenth century to the year 2021. Thanks to colonies preserved in some Italian historical museum collections, originating from the western basin, it was possible to date the putative first documented record of P. physalis of the Mediterranean Sea in 1850. The dataset shows some massive strandings that occurred in localities of the Strait of Gibraltar and Alboran Sea, the area of the entrance, from the Atlantic toward the Mediterranean waters, and the starting point from where the species spread toward the western and central basin. Physalia physalis does not reach the eastern area of the Mediterranean Sea. As the records of this species from the Italian maritime regions were abundant in the summertime and considering the danger related to contact with humans, they were subdivided into three categories of risk according to the months of occurrence. These categories were created to assign a level of danger for swimmers. The increasing sightings of such a poisonous cnidarian in coastal waters can represent a risk to human health, and a threat to all those activities linked to the marine tourism sector. The overview given goes beyond scientific purposes and aims to reach society and public administrators. The involvement of citizens and touristic structures for the early detection of P. physalis can play a key role in preventing encounters with the species, allowing marine tourist facilities to operate within a range of reasonable security.Open Access Funding provided by Università degli Studi di Palermo. This work was carried out within the framework of the Ph.D. project of RB funded by Università degli Studi di Palermo. This research was also supported by the Project Agreement “Sistema de Observación y Predicción de Medusas en el Mar Balear” among Govern des Illes Balears, SOCIB and CSIC (Disposición 15052, BOE núm. 310, 2020).Peer reviewe

Sicilian Populations of <i>Capparis spinosa</i> L. and <i>Capparis orientalis</i> Duhamel as Source of the Bioactive Flavonol Quercetin

The genus Capparis is a taxon of difficult delimitation that has several species and ecotypes due to its wide heterogeneity, its extreme phenotypic diversity, and the presence of intermediate forms linked to hybridization phenomena. The Sicilian territory hosts numerous wild and cultivated populations of two spp. Capparis spinosa L. and Capparis orientalis Duhamel, which are ecologically and morphologically distinct. The caper has considerable interest and economic value for its medicinal properties, culinary uses, and cultivation characteristics. It is one of the foods with the highest quercetin content. Quercetin is a flavonol with antioxidant, anti-inflammatory, and immunostimulant properties. Recently, patents and clinical studies have highlighted the inhibitory effect of this compound against several SARS-CoV-2 enzymes (MPro, PLPro, and RdRp). Therefore, the aim of this study was to quantify the amount of quercetin in C. spinosa and C. orientalis by LC-ESI/QTrap/MS/MS and to correlate it with the pedoclimatic features. The results obtained showed that quercetin is more abundant in C. orientalis than in C. spinosa. The highest values of quercetin were recorded in C. orientalis flowers, leaves, and flower buttons of volcanic islands with southwest and east warm exposures. In conclusion, the data acquired can provide a good basis for further scientific investigations to support the identification of possible ecotypes as a source of quercetin for food or pharmaceutical purposes