The relevant effect of marine salt and epiphytes on Posidonia oceanica waste pyrolysis: Removal of SO2/HCl emissions and promotion of O/HCOOH formation.

Significant quantities of Posidonia oceanica deposit on some beaches and coastlines every year, which generates high costs associated with the disposal of this waste. Pyrolysis may be an adequate way for its valorization. However, it would imply to know how the process takes place and if the removal of its natural detrital inorganic matter (epiphytes, marine salt and sand) is necessary, which are the objectives of this research. Pyrolysis by thermogravimetry-mass spectrometry was carried out on both the washed and unwashed samples. During this waste pyrolysis, the following occurs: (i) the high alkali metal chloride content promotes fragmentation reactions of carbohydrates and O formation, which increases HCOOH intensities at temperatures between 250 and 360 °C; (ii) from 500 °C to 650 °C, Fe2O3 and decomposition of carbonates seem to be involved in reactions that produce O release and steam and CO2 reforming of hydrocarbons and oxygenated organic compounds with H2 generation; (iii) from 650 °C to 750 °C, Fe2O3, high alkali metal content and carbonate decomposition generate char gasification, an increase in O release, SO2 capture and HCOOH formation. In general, the abundance of inorganic matter (chlorides, carbonates, etc.) minimizes the release of various compounds during pyrolysis, including SO2 and HCl, while increasing HCOOH production. Thus, this high content of inorganic matter may represent an advantage for its pyrolysis, producing value-added chemical products with a reduced environmental impact. Therefore, this study may be the starting point for defining the optimal pyrolysis conditions for this waste valorisation.

The power of Posidonia oceanica meadows to retain microplastics and the consequences on associated macrofaunal benthic communities.

In the coastal environment, a large amount of microplastics (MPs) can accumulate in the sediments of seagrass beds. However, the potential impact these pollutants have on seagrasses and associated organisms is currently unknown. In this study, we investigated the differences in MPs abundance and composition (i.e., shape, colour and polymer type) in marine sediments collected at different depths (-5 m, -15 m, -20 m) at two sites characterized by the presence of Posidonia oceanica meadows and at one unvegetated site. In the vegetated sites, sediment samples were collected respectively above and below the upper and lower limits of the meadow (-5 m and -20 m), out of the P. oceanica meadow, and in the central portion of the meadow (-15 m). By focusing on the central part of the meadow, we investigated if the structural features (i.e. shoots density and leaf surface) can affect the amount of MPs retained within the underlying sediment and if these, in turn, can affect the associated benthic communities. Results showed that the number of MPs retained by P. oceanica meadows was higher than that found at the unvegetated site, showing also a different composition. In particular, at vegetated sites, we observed that MPs particles were more abundant within the meadow (at - 15 m), compared to the other depths, on unvegetated sediment, with a dominance of transparent fragments of polypropylene (PP). We observed that MPs entrapment by P. oceanica was accentuated by the higher shoots density, while the seagrass leaf surface did not appear to have any effect. Both the abundance and richness of macrofauna associated with P. oceanica rhizomes appear to be negatively influenced by the MPs abundance in the sediment. Overall, this study increases knowledge of the potential risks of MPs accumulation in important coastal habitats such as the Posidonia oceanica meadows.

Effects of acidification on the biogeochemistry of unvegetated and seagrass marine sediments.

Many studies addressed ocean acidification (OA) effects on marine life, whereas its effects on sedimentary organic matter (OM) have received less attention. We investigated differences in OM features in sediments from unvegetated and seagrass (Posidonia oceanica) beds in a shallow hydrothermal area (Aeolian Archipelago, Mediterranean Sea), under natural (8.1-8.0) and acidified (7.8-7.9) conditions. We show that a pH difference of -0.3 units have minor effects on OM features in unvegetated sediments, but relevant consequences within acidified seagrass meadows, where OM quantity and nutritional quality are lower than those under natural pH conditions. Effects of acidified conditions on OM biogeochemistry vary between unvegetated and seagrass sediments, with lower C degradation rates and longer C turnover time in the former than in the latter. We conclude that OA, although with effects not consistent between unvegetated and vegetated sediments, can affect OM quantity, composition, and degradation, thus having possible far-reaching consequences for benthic trophic webs.

Undisturbed Posidonia oceanica meadows maintain the epiphytic bacterial community in different environments.

Seagrasses harbour different and rich epiphytic bacterial communities. These microbes may establish intimate and symbiotic relationships with the seagrass plants and change according to host species, environmental conditions, and/or ecophysiological status of their seagrass host. Although Posidonia oceanica is one of the most studied seagrasses in the world, and bacteria associated with seagrasses have been studied for over a decade, P. oceanica's microbiome remains hitherto little explored. Here, we applied 16S rRNA amplicon sequencing to explore the microbiome associated with the leaves of P. oceanica growing in two geomorphologically different meadows (e.g. depth, substrate, and turbidity) within the Limassol Bay (Cyprus). The morphometric (leaf area, meadow density) and biochemical (pigments, total phenols) descriptors highlighted the healthy conditions of both meadows. The leaf-associated bacterial communities showed similar structure and composition in the two sites; core microbiota members were dominated by bacteria belonging to the Thalassospiraceae, Microtrichaceae, Enterobacteriaceae, Saprospiraceae, and Hyphomonadaceae families. This analogy, even under different geomorphological conditions, suggest that in the absence of disturbances, P. oceanica maintains characteristic-associated bacterial communities. This study provides a baseline for the knowledge of the P. oceanica microbiome and further supports its use as a putative seagrass descriptor.

Sustainable Exploitation of Posidonia oceanica Sea Balls (Egagropili): A Review.

Posidonia oceanica (L.) Delile is the main seagrass plant in the Mediterranean basin that forms huge underwater meadows. Its leaves, when decomposed, are transported to the coasts, where they create huge banquettes that protect the beaches from sea erosion. Its roots and rhizome fragments, instead, aggregate into fibrous sea balls, called egagropili, that are shaped and accumulated by the waves along the shoreline. Their presence on the beach is generally disliked by tourists, and, thus, local communities commonly treat them as waste to remove and discard. Posidonia oceanica egagropili might represent a vegetable lignocellulose biomass to be valorized as a renewable substrate to produce added value molecules in biotechnological processes, as bio-absorbents in environmental decontamination, to prepare new bioplastics and biocomposites, or as insulating and reinforcement materials for construction and building. In this review, the structural characteristics, and the biological role of Posidonia oceanica egagropili are described, as well as their applications in different fields as reported in scientific papers published in recent years.

New Bioactive Peptides from the Mediterranean Seagrass Posidonia oceanica (L.) Delile and Their Impact on Antimicrobial Activity and Apoptosis of Human Cancer Cells.

The demand for new molecules to counter bacterial resistance to antibiotics and tumor cell resistance is increasingly pressing. The Mediterranean seagrass Posidonia oceanica is considered a promising source of new bioactive molecules. Polypeptide-enriched fractions of rhizomes and green leaves of the seagrass were tested against Gram-positive (e.g., Staphylococcus aureus, Enterococcus faecalis) and Gram-negative bacteria (e.g., Pseudomonas aeruginosa, Escherichia coli), as well as towards the yeast Candida albicans. The aforementioned extracts showed indicative MIC values, ranging from 1.61 µg/mL to 7.5 µg/mL, against the selected pathogens. Peptide fractions were further analyzed through a high-resolution mass spectrometry and database search, which identified nine novel peptides. Some discovered peptides and their derivatives were chemically synthesized and tested in vitro. The assays identified two synthetic peptides, derived from green leaves and rhizomes of P. oceanica, which revealed interesting antibiofilm activity towards S. aureus, E. coli, and P. aeruginosa (BIC50 equal to 17.7 µg/mL and 70.7 µg/mL). In addition, the natural and derivative peptides were also tested for potential cytotoxic and apoptosis-promoting effects on HepG2 cells, derived from human hepatocellular carcinomas. One natural and two synthetic peptides were proven to be effective against the "in vitro" liver cancer cell model. These novel peptides could be considered a good chemical platform for developing potential therapeutics.

Enhanced Streptomyces roseochromogenes melanin production by using the marine renewable source Posidonia oceanica egagropili.

Since the possibility to biotechnologically produce melanin by Streptomycetes using plant biomass has been so far poorly investigated, Posidonia oceanica egagropili, a marine waste accumulating along the Mediterranean Sea coasts, was explored as a renewable source to enhance extracellular melanin production by Streptomyces roseochromogenes ATCC 13400. Therefore, different amounts of egagropili powder were added to a culture medium containing glucose, malt extract, and yeast extract, and their effect on the melanin biosynthesis was evaluated. A 2.5 g·L-1 supplementation in 120-h shake flask growths at 26 °C, at pH 6.0 and 250 rpm, was found to enhance the melanin production up to 3.94 ± 0.12 g·L-1, a value 7.4-fold higher than the control. Moreover, 2-L batches allowed to reach a concentration of 9.20 ± 0.12 g·L-1 in 96 h with a productivity of 0.098 g·L-1·h-1. Further studies also demonstrated that the melanin production enhancement was due to the synergistic effect of both the lignin carbohydrate complex and the holocellulose components of the egagropili. Finally, the pigment was purified from the broth supernatant by acidic precipitation and reversed-phase chromatography, characterized by UV absorbance and one- and two-dimensional NMR, and also tested for its chemical, antioxidant, and photo-protective properties. KEY POINTS: • S. roseochromogenes ATCC 13400 produces extracellular soluble melanin. • Egagropili added to the growth medium enhances melanin production and productivity. • Both the lignin carbohydrate complex and the holocellulose egagropili components influence the melanin biosynthesis.

Environmental settings of seagrass meadows control rare earth element distribution and transfer from soil to plant compartments.

The role of seagrass meadows in the cycling and accumulation of rare earth elements and yttrium (REEY) is unknown. Here, we measured the concentration of REEY in the different compartments of Halodule wrightii (shoots, rhizomes, and roots) and soils in seagrass meadows near sandy beaches, mangroves, and coral reefs in the Todos os Santos Bay, Brazil. We provide data on the accumulation dynamics of REEY in seagrass compartments and demonstrate that plant compartments and soil properties determine accumulation patterns. The ∑REEY in soils were ~1.7-fold higher near coral reefs (93.0 ±â€¯5.61 mg kg-1) than near mangrove sites (53.9 ±â€¯31.5 mg kg-1) and were slightly higher than in sandy beaches (81.7 ±â€¯49.1 mg kg-1). The ∑REEY in seagrasses varied between 35.4 ±â€¯28.1 mg kg-1 near coral reefs to 59.2 ±â€¯21.3 mg kg-1 near sandy beaches, respectively. The ∑REE bioaccumulation factor (BAF) was highest in seagrass roots near sandy beaches (BAF = 0.67 ±â€¯0.48). All values of ∑REE translocation are <1, indicating inefficient translocation of REE from roots to rhizome to shoot. PAAS normalized REE was enriched in light REE (LREE) over heavy REE (HREE). The REEY accumulation in Halodule wrightii revealed a low potential of the seagrass to act as a sink for these elements. However, their bioavailability and potential uptake may change with soil properties. Our results serve as a basis for a better understanding of REE biogeochemical cycling and its fate in the marine environment. REE have experienced increased use as they are central to new technologies revealing an urgent need for further investigations of potential impacts on coastal ecosystems.

Early evidence of the impacts of microplastic and nanoplastic pollution on the growth and physiology of the seagrass Cymodocea nodosa.

Microplastics (MPs) and nanoplastics (NPs) are ubiquitous in natural habitats and the risks their presence poses to marine environments and organisms are of increasing concern. There is evidence that seagrass meadows are particularly prone to accumulate plastic debris, including polystyrene particles, but the impacts of this pollutant on seagrass performance are currently unknown. This is a relevant knowledge gap as seagrasses provide multiple ecosystem services and are declining globally due to anthropogenic impact and climate-change-related stressors. Here, we explored the potential effects of a 12 day-exposure of seagrasses to one concentration (68 µg/L) of polystyrene MPs and NPs on the growth, oxidative status, and photosynthetic efficiency of plants using the foundation species Cymodocea nodosa as a model. Among plant organs, adventitious roots were particularly affected by MPs and NPs showing complete degeneration. The number of leaves per shoot was lower in MPs- and NPs-treated plants compared to control plants, and leaf loss exceeded new leaf production in MPs-treated plants. MPs also reduced photochemical efficiency and increased pigment content compared to control plants. Shoots of NPs-treated plants showed a greater oxidative damage and phenol content than those of control plants and MPs-treated plants. Biochemical data about oxidative stress markers were consistent with histochemical results. The effects of MPs on C. nodosa could be related to their adhesion to plant surface while those of NPs to entering tissues. Our study provides the first experimental evidence of the potential harmful effects of MPs/NPs on seagrass development. It also suggests that the exposure of seagrasses to MPs/NPs in natural environments could have negative consequences on the functioning of seagrass ecosystems. This stresses the importance of implementing cleaning programs to remove all plastics already present in marine habitats as well as of undertaking specific actions to prevent the introduction of these pollutants within seagrass meadows.

Low-cost Posidonia oceanica bio-adsorbent for efficient removal of antibiotic oxytetracycline from water.

The presence of antibiotics as micro-contaminants in the water and aqueous environments is a health concern to humans and the ecosystem. Therefore, their elimination by adsorption to available and cheap materials in water treatment plants is a research topic of high relevance. The present paper reports on the adsorption behavior of oxytetracycline on a bio-adsorbent prepared from Posidonia oceanica; an abundant Mediterranean biomass. Characterization of the pretreated Posidonia biomaterial was achieved using several analyses such as Boehm acid-base titration method, pHPZC determination, and analysis techniques (FTIR, 13C CP-MAS NMR, optical microscopy, and TGA). The pHPZC occurred around pH 2.11. Posidonia biomaterial showed a fast and high uptake rate throughout the adsorption process, which is a definite advantage for analytical applications such as water decontamination. The experimental kinetic data fitted very rightly the pseudo-second-order kinetic model and the equilibrium uptake can adopt the bi-Langmuir isotherm model for all studied pH values which assumes adsorptions at the two localized sites. Maximum adsorption capacities of 11.8 mg∙g-1 and 4.4 mg∙g-1 for the two adsorption sites are reached at pH 6. The oxytetracycline adsorption process onto Posidonia bio-adsorbent is spontaneous (ΔadsG0 < 0), exothermic (ΔadsH0 < 0), and entropically favorable (ΔadsS0 > 0). The effect of pH on adsorption behavior and the thermodynamic parameters of adsorption are consistent with a possible origin of adsorption of oxytetracycline by means of hydrogen bonding interactions between surface hydroxyl and phenolic groups of the biomaterial and oxytetracycline. The proposed green and environmentally friendly biomaterial offers potential benefits as a bio-adsorbent in the remediation of aquatic environments contaminated by various organic materials.

Profiling the cell walls of seagrasses from A (Amphibolis) to Z (Zostera).

BACKGROUND: The polyphyletic group of seagrasses shows an evolutionary history from early monocotyledonous land plants to the marine environment. Seagrasses form important coastal ecosystems worldwide and large amounts of seagrass detritus washed on beaches might also be valuable bioeconomical resources. Despite this importance and potential, little is known about adaptation of these angiosperms to the marine environment and their cell walls. RESULTS: We investigated polysaccharide composition of nine seagrass species from the Mediterranean, Red Sea and eastern Indian Ocean. Sequential extraction revealed a similar seagrass cell wall polysaccharide composition to terrestrial angiosperms: arabinogalactans, pectins and different hemicelluloses, especially xylans and/or xyloglucans. However, the pectic fractions were characterized by the monosaccharide apiose, suggesting unusual apiogalacturonans are a common feature of seagrass cell walls. Detailed analyses of four representative species identified differences between organs and species in their constituent monosaccharide composition and lignin content and structure. Rhizomes were richer in glucosyl units compared to leaves and roots. Enhalus had high apiosyl and arabinosyl abundance, while two Australian species of Amphibolis and Posidonia, were characterized by high amounts of xylosyl residues. Interestingly, the latter two species contained appreciable amounts of lignin, especially in roots and rhizomes whereas Zostera and Enhalus were lignin-free. Lignin structure in Amphibolis was characterized by a higher syringyl content compared to that of Posidonia. CONCLUSIONS: Our investigations give a first comprehensive overview on cell wall composition across seagrass families, which will help understanding adaptation to a marine environment in the evolutionary context and evaluating the potential of seagrass in biorefinery incentives.

Specialized compounds across ontogeny in the seagrass Posidonia oceanica.

Differences in phenolic composition across different ontogenic stages can be crucial in determining the interaction outcomes between plants and their surrounding biotic environment. In seagrasses, specific phenolic compounds have rarely been analyzed and remain unexplored in ontogenic stages other than non-reproductive adults. Furthermore, it is generally accepted that plants would prioritize defense (e.g., through increased phenolic content) on tissues or stages that are critical for plant fitness but how this affects nutritional quality or plant resources has been scarcely explored. We analyzed how phenolic composition, N and C content and carbohydrate resources varied among different life stages (i.e. old and young leaves of reproductive and non-reproductive plants, and leaves of seedlings) in the seagrass Posidonia oceanica. We identified five phenolic compounds, whose structures were established as hydroxycinnamate esters of tartaric acid. Also, our results show that in all examined ontogenic stages phenolic compounds have the same qualitative composition but inflorescences exhibit higher contents than vegetative tissues. We did not find a reduction in stored resources in reproductive plants, pointing to some kind of compensatory mechanism in the production or storage of resources. In contrast, seedlings seemed to have less phenolic compounds than reproductive plants, perhaps due to limited resources available to allocate to phenolic production. Our results demonstrate how different ontogenic stages change their investment in specialized phenolic compounds prioritizing different functions according to the needs and limitations of that stage.

Lignin/Carbohydrate Complex Isolated from Posidonia oceanica Sea Balls (Egagropili): Characterization and Antioxidant Reinforcement of Protein-Based Films.

A lignin fraction (LF) was extracted from the sea balls of Posidonia oceanica (egagropili) and extensively dialyzed and characterized by FT-IR and NMR analyses. LF resulted water soluble and exhibited a brownish-to-black color with the highest absorbance in the range of 250-400 nm, attributed to the chromophore functional groups present in the phenylpropane-based polymer. LF high-performance size exclusion chromatography analysis showed a highly represented (98.77%) species of 34.75 kDa molecular weight with a polydispersity index of 1.10 and an intrinsic viscosity of 0.15. Quantitative analysis of carbohydrates indicated that they represented 28.3% of the dry weight of the untreated egagropili fibers and 72.5% of that of LF. In particular, eight different monosaccharides were detected (fucose, arabinose, rhamnose, galactose, glucose, xylose, glucosamine and glucuronic acid), glucuronic acid (46.6%) and rhamnose (29.6%) being the most present monosaccharides in the LF. Almost all the phenol content of LF (113.85 ± 5.87 mg gallic acid eq/g of extract) was water soluble, whereas around 22% of it consisted of flavonoids and only 10% of the flavonoids consisted of anthocyanins. Therefore, LF isolated from egagropili lignocellulosic material could be defined as a water-soluble lignin/carbohydrate complex (LCC) formed by a phenol polymeric chain covalently bound to hemicellulose fragments. LCC exhibited a remarkable antioxidant activity that remained quite stable during 6 months and could be easily incorporated into a protein-based film and released from the latter overtime. These findings suggest egagropili LCC as a suitable candidate as an antioxidant additive for the reinforcement of packaging of foods with high susceptibility to be deteriorated in aerobic conditions.

Multifunctional cellulosic aerogels from Posidonia oceanica waste biomass with antioxidant properties for meat preservation.

Posidonia oceanica waste biomass has been valorized to develop bioactive multifunctional cellulosic aerogels (HCAG) by simpler and greener protocols. Hydrophobization of cellulosic aerogels was achieved through PLA coating, while bioactivity was imparted by the incorporation of hydrophilic (E2) and hydrophobic extracts (E3) produced from the same biomass. The incorporation of extracts led to denser aerogels, with less porous structures. These aerogels showed outstanding water and oil sorption capacities (1500-1900%), being able to release the adsorbed liquid almost completely after 7 days. Interestingly, all the aerogels showed a positive inhibition effect (23-91%) on the ß-carotene bleaching assay. Moreover, the aerogels loaded with extracts, especially when combining E2 and E3, were able to reduce the oxidation of lipids and oxymyoglobin in red meat after 10 days of storage. This evidences the potential of these multifunctional aerogels as bioactive adsorbing pads to preserve the quality of fresh packaged foods.

Seagrass Posidonia is impaired by human-generated noise.

The last hundred years have seen the introduction of many sources of artificial noise in the sea environment which have shown to negatively affect marine organisms. Little attention has been devoted to how much this noise could affect sessile organisms. Here, we report morphological and ultrastructural changes in seagrass, after exposure to sounds in a controlled environment. These results are new to aquatic plants pathology. Low-frequency sounds produced alterations in Posidonia oceanica root and rhizome statocysts, which sense gravity and process sound vibration. Nutritional processes of the plant were affected as well: we observed a decrease in the number of rhizome starch grains, which have a vital role in energy storage, as well as a degradation in the specific fungal symbionts of P. oceanica roots. This sensitivity to artificial sounds revealed how sound can potentially affect the health status of P. oceanica. Moreover, these findings address the question of how much the increase of ocean noise pollution may contribute in the future to the depletion of seagrass populations and to biodiversity loss.

Inhibition of MMP2-PEX by a novel ester of dihydroxy cinnamic and linoleic acid from the seagrass Cymodocea serrulata.

Matrix metalloproteinases (MMPs) are pivotal for cancer cell migration and metastasis which are generally over-expressed in such cell types. Many drugs targeting MMPs do so by binding to the conserved catalytic domains and thus exhibit poor selectivity due to domain-similarities with other proteases. We report herein the binding of a novel compound [3-(E-3,4-dihydroxycinnamaoyloxyl)-2-hydroxypropyl 9Z, 12Z-octadeca-9, 12-dienoate; Mol. wt: 516.67 Da], (C1), isolated from a seagrass, Cymodocea serrulata to the unconserved hemopexin-like (PEX) domain of MMP2 (- 9.258 kcal/mol). MD simulations for 25 ns, suggest stable ligand-target binding. In addition, C1 killed an ovarian cancer cell line, PA1 at IC50: 5.8 µM (lesser than Doxorubicin: 8.6 µM) and formed micronuclei, apoptotic bodies and nucleoplasmic bridges whilst causing DNA laddering, S and G2/M phase dual arrests and MMP disturbance, suggesting intrinsic apoptosis. The molecule increased mRNA transcripts of BAX and BAD and down-regulated cell survival genes, Bcl-xL, Bcl-2, MMP2 and MMP9. The chemical and structural details of C1 were deduced through FT-IR, GC-MS, ESI-MS, 1H and 13C NMR [both 1D and 2D] spectra.

Halodule pinifolia (Seagrass) attenuated lipopolysaccharide-, carrageenan-, and crystal-induced secretion of pro-inflammatory cytokines: mechanism and chemistry.

OBJECTIVES: The study aimed to explore the anti-inflammatory effect, underlying mechanism, and chemistry of Halodule pinifolia extract. METHODS: The ethyl acetate (EHP) and methanol (MHP) extracts of Halodule pinifolia were screened for pro-inflammatory cytokine inhibition effect under various in vitro (LPSand crystal-induced inflammation) and in vivo models (LPS-induced endotoxaemia model, carrageenan-induced paw oedema model, and oxalate-induced renal nephropathy model of inflammation). The effect of EHP on the expression of inflammatory markers using western blot analysis (in vitro) was investigated. Chemical constituents of bioactive EHP were isolated through chromatography and characterised using NMR spectroscopy. Furthermore, EHP was standardised for rosmarinic acid, vanillic acid, and ethyl protocatechuate using HPLC. Also, total phytosterols, phenolic, and flavonoid content of EHP were determined by UV spectroscopy. KEY FINDINGS: EHP was comparatively more effective than MHP in inhibiting cytokines secretions under LPS-induced in vitro models. Furthermore, EHP was screened under endotoxaemia in vivo model, EHP (250 mg/kg) reduced plasma IL-6, TNF-α, and IL-1ß levels by 88.3%, 78.2%, and 74.5%, respectively. In the carrageenan-induced oedema model, EHP (200 mg/kg) reduced paw volume and release of TNF-α (69.3%) and IL-1ß (43.1%). EHP (200 mg/kg) further controlled renal nephropathy by inhibiting plasma IL-1ß and BUN levels. Also, a significant reduction of mRNA expressions of TNF-α and IL-1ß and KIM-1 in renal tissues was observed. Through western blot, EHP was identified to regulate the expression of pro-form as well as mature-form of IL-1ß and caspase-1. EHP constituted rosmarinic acid (RA), vanillic acid (VA), ethyl protocatechuate (EP), sitosterol, stigmasterol, campesterol, and dihydrobrassicasterol. It was determined that 4.6 mg/g of RA, 2.92 mg/g of VA, 0.76 mg/g of EP, 21.7 mg/g of total phenolics, 29.8 mg/g of total flavonoids, and 48.2 mg/g of total phytosterols were present in dry EHP. The presence of anti-inflammatory constituents such as RA, VA, and PE in EHP corroborated the in vitro and in vivo anti-inflammatory activity of EHP. CONCLUSION: The anti-inflammatory property of EHP and its action through attenuation of pan-cytokines suggest that it can be developed into an oral pharmaceutical drug.

Alisol A is potentially therapeutic in human breast cancer cells.

Recent developments in breast cancer therapy have significantly improved patient survival rate; however, recurrence remains a major problem. Systemic treatment of breast cancer with available therapies is not curative. Natural products can be potentially used for treating cancer. Recently, a wide range of pharmacological activities has been reported for Alismatis Rhizoma, a popular traditional Chinese medicine. However, the mechanisms via which its compounds act on breast cancer remain unclear. The present study aimed to investigate the potential of natural therapeutic agents from Alismatis Rhizoma for treating breast cancer. Human breast cancer MDA­MB­231 cells were treated with four main protostane triterpenes from Alismatis Rhizoma, including alisol A, alisol A 24­acetate, alisol B and alisol B 23­acetate. Among these, alisol A significantly inhibited cell viability. Alisol A induced cell apoptosis, G1 phase cell cycle arrest, autophagy, and intracellular reactive oxygen species (ROS) generation in MDA­MB­231 cells. The number of APE1­/γH2AX­/LC3­II positive cells was also significantly higher compared with that of negative control cells. All these results were dose­dependent. Cleaved caspase­3, cleaved caspase 9, Bcl­2, and p­p38 expression indicated cell apoptosis after alisol A treatment. The changes in cyclin A and cyclin D1 expression was associated with cell cycle arrest upon alisol A treatment. Furthermore, LC3­II expression upon alisol A treatment was indicative of autophagy. Alisol A treatment can induce autophagy­dependent apoptosis in human breast cancer cells via induction of ROS and DNA damage. Thus, Alisol A might serve as a new therapeutic agent against breast cancer.

In vitro anti-glycation activity of the marine plant Posidonia oceanica (L.) Delile.

ETHNOPHARMACOLOGICAL RELEVANCE: The marine plant Posidonia oceanica (L.) Delile is traditionally used by villagers of the west coast of Anatolia as a remedy for diabetes and hypertension. AIM OF THE STUDY: The aim of this study was to explore the role of the P. oceanica hydroalcoholic leaves extract (POE) against human serum albumin glycation. MATERIAL AND METHODS: Advanced glycation end products (AGEs) were obtained with the albumin-glucose in vitro assay. The AGEs intrinsic fluorescence intensity and the electrophoretic migration under native conditions allowed us to verify the effective glycation of albumin. The presence of POE during glycation process was intended to evaluate its anti-glycation role. RESULTS: POE exhibited a strong in vitro anti-glycation ability which occurred independently from its known antioxidant property. CONCLUSIONS: Overall, the antidiabetic, antioxidant, anti-inflammatory and anti-glycation properties of POE could be exploited as an effective tool against diabetes and related complications.

Sink trap: duckweed and dye attractant reduce mosquito populations.

Duckweeds, such as Lemna minor Linnaeus (Alismatales: Lemnaceae), are common in aquatic habitats and have been suggested to reduce larval mosquito survivorship via mechanical and chemical effects. Furthermore, pond dyes are used increasingly in aquatic habitats to enhance their aesthetics, although they have been shown to attract mosquito oviposition. The present study examined the coupled effects of L. minor and black pond dye on the oviposition selectivity of Culex pipiens Linnaeus (Diptera: Culicidae) mosquitoes in a series of laboratory choice tests. Subsequently, using outdoor mesocosms, the combined influence of duckweed and pond dye on mosquito abundances in aquatic habitats was quantified. Mosquitoes were strongly attracted to duckweed, and oviposited significantly greater numbers of egg rafts in duckweed-treated water compared with untreated controls, even when the duckweed was ground. The presence of pond dye interacted with the duckweed and further enhanced positive selectivity towards duckweed-treated water. The presence of duckweed caused significant and sustained reductions in larval mosquito numbers, whereas the relative effects of dye were not evident. The use of floating aquatic plants such as duckweed, combined with dye, may help reduce mosquito populations via the establishment of population sinks, characterized by high rates of oviposition coupled with high levels of larval mortality.