Fatty Acids in Waste Tissues: The Nutraceutical Value of Gonads and Livers from the Moroccan Hypophthalmichthys molitrix and Cyprinus carpio Fishes.

Fishes are an important component of human nutrition, mainly acting as source of essential fatty acids in the prevention of cardiovascular disorders. The increase in their consumption has led to a growth of fishes waste; therefore, the disposal and recycling of waste has become a key issue to address, in accordance with circular economy principles. The Moroccan Hypophthalmichthys molitrix and Cyprinus carpio fishes, living in freshwater and marine environments, were collected at mature and immature stages. The fatty acid (FA) profiles of liver and ovary tissues were investigated by GC-MS and compared with edible fillet tissues. The gonadosomatic index, the hypocholesterolemic/hypercholesterolemic ratio, and the atherogenicity and thrombogenicity indexes were measured. Polyunsaturated fatty acids were found to be abundant in the mature ovary and fillet of both species, with a polyunsaturated fatty acids/saturated fatty acids ratio ranging from 0.40 to 1.06 and a monounsaturated fatty acids/polyunsaturated fatty acids ratio between 0.64 and 1.84. Saturated fatty acids were found to be highly abundant in the liver and gonads of both species (range 30-54%), as well as monounsaturated fatty acids (range 35-58%). The results suggested that the exploitation of fish wastes, such as the liver and ovary, may represent a sustainable strategy for the achievement of high value-added molecules with nutraceutical potential.

Taste Compounds and Polyphenolic Profile of Tomato Varieties Cultivated with Beneficial Microorganisms: A Chemical Investigation on Nutritional Properties and Sensory Qualities.

There is a strong need to develop eco-sustainable agricultural techniques to improve crop yields while preserving biomolecule contents and reducing the adverse environmental impact of agro-chemicals. The use of microorganisms in agriculture represents an attractive and innovative solution. Herein, a chemical study on the nutritional and sensory qualities of San Marzano Cirio 3 (SMC3), Corbarino (CO) and Brandywine (BW) tomato varieties cultivated with and without effective microorganisms (EM) is reported. LC-MS analysis of the methanolic extracts allowed for the identification of 21 polyphenol derivatives. In different proportions among the studied varieties, the two main polyphenols were rutin and naringenin chalcone; the latter was isolated and chemically identified by complementary HR-ESIMS/MS and NMR methods. SMC3 and CO were richer in naringenin chalcone. Conversely, BW showed higher proportions of rutin; however, in all cases, the relative amounts of the two polyphenols considered together increased over the other minor components after the EM treatment. The qualitative and quantitative HPLC analyses of taste-active compounds (aspartic acid, glutamic acid, AMP and GMP) revealed a significant difference in aspartic and glutamic acids and ribonucleotide contents according to the cultivation condition (±EM), particularly in BW. This study provides chemical data in support of the use of EM green technology for the cultivation of edible agricultural products, such as tomato preserves, and may even improve nutritional and sensory qualities while safeguarding the environment.

Comparative Fatty Acid Profiling of Edible Fishes in Kuala Terengganu, Malaysia.

The aim of this study was to compare the relative nutritional benefit of edible Malaysian fishes from the coast of Terengganu in Malaysia, as well as to perform a taxonomical characterization and metal assessment. Discrimination between species was carried out by a morphological and molecular approach by evaluating the total concentrations of metals by ICP-MS analyses and the fatty acids (FA) composition using the GC-MS approach on the fish fillet tissues. The taxonomical studies detected fishes of 11 families and 13 species. The heavy metal assessment showed that all detected elements did not exceed the regulatory limit stated by Malaysian Food Regulations. The proportion of saturated fatty acids (SFA) ranged from 33 to 58.34%, followed by the polyunsaturated fatty acids (PUFA) values from 24 to 51.8%, and the lowest proportion was of monounsaturated fatty acids (MUFA), ranging from 12.7 to 35.9%. The ω-3/ω-6 PUFA and PUFA/SFA ratios were determined in the range 1.1 to 7.4 and 0.35 to 1.6, respectively. The C20:5 ω-3 and C22:6 ω-3 acids were detected at levels comparable to those found in the corresponding species from similar tropical marine ecosystems. The high FA values can be useful biochemical tools for comparing the relative nutritional benefits of these biodiverse and non-toxic edible Malaysian fishes.

Determination of flavor-potentiating compounds in different Italian tomato varieties.

Umami taste, known as appetizing sensation, is mainly imparted by monosodium glutamate (MSG, the first identified umami factor) in synergistic combination with some 5' ribonucleotides such as inosine 5'-monophosphate, IMP, guanosine 5'-monophosphate, GMP, and adenosine 5'-monophoshate, AMP. The level of free glutamic acid in tomatoes is higher than in other vegetables or fruits and increases with ripening and industrial processing. In addition, due to the presence of bioactive metabolites, tomatoes and tomato-based products are among the most consumed healthy food items. The levels of the major umami compounds of tomato, that is, glutamate and 5'-ribonucleotides (GMP and AMP) were assessed in different parts (skin, outer flesh, and inner pulp) of known tomato varieties from southern Italy: San Marzano Originale, San Marzano 245, Black Tomato, Corbarino Corbara, Corbarino Nocera, and Superpomodoro (tomato hybrid). Such varieties were also investigated for their antioxidant properties through DMPD, DPPH, and ABTS assays, with San Marzano Originale showing the highest antioxidant power both in lipophilic and methanolic fractions. The concentration of umami compounds in tomato differs with the part of the fruit analyzed and is greatly dependent on the variety, being Corbarino Nocera the cultivar richest in glutamate and Superpomodoro in ribonucleotides. As for nutritional aspect, results confirm the great nutraceutical feature of San Marzano tomato, the most known variety used in industrial processes. PRACTICAL APPLICATIONS: This study was planned to develop a method to quantify the major umami compounds that strongly influence the organoleptic properties of many different tomato varieties. It is known that the sensory quality of fruits and vegetables is an important factor in consumer's choice. The analytical methods described here enabled the evaluation of the glutamate and 5'-ribonucleotides contents in six selected varieties of tomato from Campania region, and can be easily used to determine the sensory profile of commercial varieties, for example, those perceived as not very tasteful by consumers.

Extremophilic Natrinema versiforme Against Pseudomonas aeruginosa Quorum Sensing and Biofilm.

Pseudomonas aeruginosa is an opportunistic pathogen that causes high morbidity and mortality rates due to its biofilm form. Biofilm formation is regulated via quorum sensing (QS) mechanism and provides up to 1000 times more resistance against conventional antibiotics. QS related genes are expressed according to bacterial population density via signal molecules. QS inhibitors (QSIs) from natural sources are widely studied evaluating various extracts from extreme environments. It is suggested that extremely halophilic Archaea may also produce QSI compounds. For this purpose, we tested QS inhibitory potentials of ethyl acetate extracts from cell free supernatants and cells of Natrinema versiforme against QS and biofilm formation of P. aeruginosa. To observe QS inhibition, all extracts were tested on P. aeruginosa lasB-gfp, rhlA-gfp, and pqsA-gfp biosensor strains and biofilm inhibition was studied using P. aeruginosa PAO1. According to our results, QS inhibition ratios of cell free supernatant extract (CFSE) were higher than cell extract (CE) on las system, whereas CE was more effective on rhl system. In addition, anti-biofilm effect of CFSE was higher than CE. Structural analysis revealed that the most abundant compound in the extracts was trans 4-(2-carboxy-vinyl) benzoic acid.

Comparative Correlation Between Chemical Composition and Cytotoxic Potential of the Coral-Associated Fungus Aspergillus sp. 2C1-EGY Against Human Colon Cancer Cells.

Cancer is a leading cause of death in several countries. In the search for new anticancer drugs, marine organisms have played an important role in the discovery of lead compounds and the development of new pharmaceuticals for their wide diversity of chemical structures and biological activities. In the present study, the cytotoxicity on colorectal cancer cells HCT116 exerted by marine fungus Aspergillus sp. 2C1-EGY extracts associated with the soft coral Sinularia sp. was investigated; the sub-fractions Fr 2c and Fr 2d had significantly high cytotoxic activity (88 and 85%, respectively). Moreover, the major hexadecanoic, octadecanoic, and octadecenoic acids as well as their methyl esters were isolated. GC/MS analysis revealed the identification of 46 major and minor compounds, from which 19 saturated and unsaturated fatty acids and eight fatty acid esters were identified.

Antioxidant activity and bioactive compound contents before and after in vitro digestion of new tomato hybrids.

BACKGROUND: The antioxidant properties and bioactive compound contents of fresh new tomato hybrids before and after in vitro digestion were investigated. To this aim, the antioxidant activities of lipophilic, hydrophilic and polyphenolic extracts of tomato hybrids were determined by ABTS (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)), DMPD (N,N-dimethyl-p-phenylenediamine dihydrochloride) and DPPH (2,2-diphenyl-1-picrylhydrazyl) methods respectively, while the bioactive compound contents were estimated via Folin-Ciocalteu (polyphenols), pH differential (anthocyanins) and high-performance liquid chromatography (lycopene and ß-carotene) methods. RESULTS: After the digestion process, a marked loss (ranging from 37 to 77%) of antioxidant capacity linked to the hydrophilic fraction was observed. In contrast, the lipophilic and methanolic fractions showed an increase in antioxidant activity (ranging from 9 to 40%) after gastric digestion, and a rapid decrease was observed after total digestion. Moreover, the presence of anthocyanins and carotenoids after simulated digestion was a notable result. CONCLUSION: The bioavailability of bioactive metabolites from nutraceutical food and their healthful properties in humans are strictly dependent on the digestion process. © 2017 Society of Chemical Industry.

The lignicolous fungus Trametes versicolor (L.) Lloyd (1920): a promising natural source of antiradical and AChE inhibitory agents.

This study aimed to determine antiradical (DPPH• and •OH) and acetylcholinesterase (AChE) inhibitory activities along with chemical composition of autochtonous fungal species Trametes versicolor (Serbia). A total of 38 phenolic compounds with notable presence of phenolic acids were identified using HPLC/MS-MS. Its water extract exhibited the highest antiradical activity against •OH (3.21 µg/mL), among the rest due to the presence of gallic, p-coumaric and caffeic acids. At the concentration of 100 µg/mL, the same extract displayed a profound AChE inhibitory activity (60.53%) in liquid, compared to donepezil (89.05%), a drug in clinical practice used as positive control. The flavonoids baicalein and quercetin may be responsible compounds for the AChE inhibitory activity observed. These findings have demonstrated considerable potential of T. versicolor water extract as a natural source of antioxidant(s) and/or AChE inhibitor(s) to be eventually used as drug-like compounds or food supplements in the treatment of Alzheimer's disease.

Investigating on the Correlation Between Some Biological Activities of Marine Sponge-Associated Bacteria Extracts and Isolated Diketopiperazines.

Marine organisms have been considered as the richest sources of novel bioactive metabolites, which can be used for pharmaceutical purposes. In the last years, the interest for marine microorganisms has grown for their enormous biodiversity and for the evidence that many novel compounds isolated from marine invertebrates are really synthesized by their associated bacteria. Nevertheless, the discovery of a chemical communication Quorum sensing (QS) between bacterial cells and between bacteria and host has gained the researchers to expand the aim of their study toward the role of bacteria associated with marine invertebrates, such as marine sponge. In the present paper, we report the evaluation of biological activities of different extracts of bacteria Vibrio sp. and Bacillus sp. associated with marine sponges Dysidea avara and Ircinia variabilis, respectively. Moreover, we evaluated the biological activities of some diketopiperazines (DKPs), previously isolated, and able to activate QS mechanism. The results showed that all extracts, fractions, and DKPs showed low scavenging activity against DPPH and superoxide anion, low cytotoxic and anti-tyrosinase activities, but no antimicrobial and acetylcholinesterase inhibitory activities. One DKP [cyclo-(trans-4-hydroxy-L-prolyl-L-leucine)] has the highest α-glucosidase inhibitory activity even than the standard acarbose.

Avarol derivatives as competitive AChE inhibitors, non hepatotoxic and neuroprotective agents for Alzheimer's disease.

Avarol is a marine sesquiterpenoid hydroquinone, previously isolated from the marine sponge Dysidea avara Schmidt (Dictyoceratida), with antiinflammatory, antitumor, antioxidant, antiplatelet, anti-HIV, and antipsoriatic effects. Recent findings indicate that some thio-avarol derivatives exhibit acetylcholinesterase (AChE) inhibitory activity. The multiple pharmacological properties of avarol, thio-avarol and/or their derivatives prompted us to continue the in vitro screening, focusing on their AChE inhibitory and neuroprotective effects. Due to the complex nature of Alzheimer's disease (AD), there is a renewed search for new, non hepatotoxic anticholinesterasic compounds. This paper describes the synthesis and in vitro biological evaluation of avarol-3'-thiosalicylate (TAVA) and thiosalycil-prenyl-hydroquinones (TPHs), as non hepatotoxic anticholinesterasic agents, showing a good neuroprotective effect on the decreased viability of SHSY5Y human neuroblastoma cells induced by oligomycin A/rotenone and okadaic acid. A molecular modeling study was also undertaken on the most promising molecules within the series to elucidate their AChE binding modes and in particular the role played by the carboxylate group in enzyme inhibition. Among them, TPH4, bearing a geranylgeraniol substituent, is the most significant Electrophorus electricus AChE (EeAChE) inhibitor (IC50 = 6.77 ± 0.24 µM), also endowed with a moderate serum horse butyrylcholinesterase (eqBuChE) inhibitory activity, being also the least hepatotoxic and the best neuroprotective compound of the series. Thus, TPHs represents a new family of synthetic compounds, chemically related to the natural compound avarol, which has been discovered for the potential treatment of AD. Findings prove the relevance of TPHs as a new possible generation of competitive AChE inhibitors pointing out the importance of the salycilic substituents on the hydroquinone ring. Since these compounds do not belong to the class of alkaloids, which are notorious for their capability to inhibit AChE while exhibiting side effects, they may constitute novel active AChE inhibitors with fewer side effects.

In vitro evaluation of cytotoxic and mutagenic activity of avarol.

The cytotoxicity of avarol, a main secondary metabolite of the Mediterranean sponge Dysidea avara, was in vitro screened by MTT assay against four human tumour cell lines. The colon HT-29 tumour cells practically showed to be the only sensitive ones towards this organic compound. No toxicity was found against the fetal lung fibroblast MRC-5 cells at the concentrations tested. In comparison with doxorubicin, used as a positive control, avarol actually exhibited at least 588-fold less toxicity towards normal MRC-5 cells. Finally, comet assay indicated that DNA fragmentation was almost fivefold higher upon the treatment with doxorubicin, compared to avarol. The obtained results have actually confirmed that avarol scaffold may contribute to development of new cytostatics inspired by nature.

Further in vitro biological activity evaluation of amino-, thio- and ester-derivatives of avarol.

The acetylcholinesterase inhibitory and/or antitumour activities of amino-, thio- and ester-derivatives of avarol selected were evaluated for the first time at in vitro conditions. Avarol-3',4'-dithioglycol (1) and avarol-4'-(3)mercaptopropionic acid (3) were shown to be the best inhibitors of the enzyme tested (0.50 µg and IC50 0.05 mM and 0.50 µg and IC50 0.12 mM, respectively), while 4'-tryptamine-avarone (9) and avarol-3'-(3)mercaptopropionic acid (2) exhibited the highest cytotoxicity against the human breast T-47D cancer cell line (IC50 0.66 µg/mL and 1.25 µg/mL, respectively). According to experimental data obtained, the sesquiterpenoid hydroquinone structure of bioactive avarol derivatives may inspire development of new pharmacologically useful substances to be used in the treatment of Alzheimer's disease and/or human breast tumour.

Effects of Industrial Processes on Antioxidant Power and Polyphenols Profile in Cherry Tomato Cultivar.

The antioxidant capacity and the polyphenolic profile of fresh and processed cherry tomatoes were analyzed with the aim of investigating the effect of industrial processes on the nutritional qualities of fruits. The results exhibited a decrease of antioxidant activity mainly in the lipophilic fraction of processed tomatoes compared with fresh products. No great difference in the antioxidant capacity was detected in the hydrophilic and methanolic extracts of fresh tomatoes and processed tomato juices. Moreover, a decrease of polyphenolic content, estimated by means of high-performance liquid chromatography analysis and Folin-Ciocalteu method, was observed in all tomato juices. The overall polyphenolic profile of both fresh and processed tomatoes did not change significantly and, among all juices, a higher polyphenolic content was detected in juice containing peels and seeds in comparison to those without. From our data on antioxidant power and the bioactive metabolite content, tomato juice could be used as a valid and easily available source of antioxidants in everyday diet to preserve human health.

Further in vitro biological activity evaluation of amino-, thio- and ester-derivatives of avarol.

The acetylcholinesterase inhibitory and/or antitumour activities of amino-, thio- and ester-derivatives of avarol selected were evaluated for the first time at in vitro conditions. Avarol-3',4'-dithioglycol (1) and avarol-4'-(3)mercaptopropionic acid (3) were shown to be the best inhibitors of the enzyme tested (0.50 µg and IC50 0.05 mM and 0.50 µg and IC50 0.12 mM, respectively), while 4'-tryptamine-avarone (9) and avarol-3'-(3)mercaptopropionic acid (2) exhibited the highest cytotoxicity against the human breast T-47D cancer cell line (IC50 0.66 µg/mL and 1.25 µg/mL, respectively). According to experimental data obtained, the sesquiterpenoid hydroquinone structure of bioactive avarol derivatives may inspire development of new pharmacologically useful substances to be used in the treatment of Alzheimer's disease and/or human breast tumour.

Further in vitro evaluation of antimicrobial activity of the marine sesquiterpene hydroquinone avarol.

This work extends in vitro screening of antimicrobial activity of the sesquiterpene hydroquinone avarol, a main secondary metabolite of the Mediterranean sponge species Dysidea avara. The antimicrobial activity was in part evaluated by microdilution method against selected bacterial and fungal strains. Additionally, the screening included evaluation of anti-quorum sensing (anti-QS) effects. At a different extent avarol was proven to be active against all the microorganisms tested (MIC 0.002-0.008 mg/mL and MBC 0.004-0.016 mg/mL; MIC 0.004-0.015 mg/mL and MFC 0.008-0.030 mg/mL; respectively). This marine natural product also showed moderate anti-QS effects, reducing Pseudomonas aeruginosa PAO1 biofilm formation (75%), its twitching and protrusions motilities as well as pyocianin production (39%). According to the best of our knowledge, this is the first report both on avarol anti Gram-negative bacterial activity and anti-QS effects.

Cyclic dipeptides produced by marine sponge-associated bacteria as quorum sensing signals.

Four bacterial strains belonging to the genera Vibrio, Pseudoalteromonas and Photobacterium were isolated from the marine sponges Dysidea avara and Geodia cynodium. A Bacillus strain was isolated from Ircinia variabilis. A screening of molecules involved in quorum sensing (QS) was carried out by TLC-overlay and a new "plate T-streak" test. To analyze quorum quenching (QQ), a plate T-streak was performed with Chromobacterium violaceum. Strains of Vibrio isolated from both marine sponges and a strain of Photobacterium isolated from G. cynodium, activated QS bioreporters. A strain of Pseudoalteromonas isolated from D. avara showed QQ activity. Finally, it is reported that cyclic dipeptides isolated from strains of Vibrio sp. and Bacillus sp. (isolated from D. avara and I. variabilis, respectively) were involved in the QS mechanism. The simultaneous presence of bacteria that showed contrasting responses in bioassays for QS signal molecule synthesis in marine sponges could add an interesting dimension to the signalling interactions which may be happening in sponges.

Further in vitro evaluation of cytotoxicity of the marine natural product derivative 4'-leucine-avarone.

The cytotoxicity of 4'-leucine-avarone, amino derivative of the sponge Dysidea avara secondary metabolite avarone, was evaluated by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide assay in vitro against seven human solid tumours for the first time. The compound tested induced dose-dependent cytotoxic response in all cancer cells showing better activity towards the lung A-549 and colon HT-29 cell lines (IC50 7.40 µM and 9.62 µM, respectively) than towards the breast adenocarcinoma ER positive MCF-7 and ER negative MDA-MB-231 cells (IC50 11.64 µM and 17.31 µM, respectively), the prostate adenocarcinoma PC-3 and epiteloid cervix carcinoma HeLa cells (IC50 14.24 µM and 15.54 µM, respectively). No toxicity was found towards the foetal lung fibroblast MRC-5 cell line at the concentrations used. According to experimental data obtained, the sesquiterpenoid quinone structure of avarone may inspire development of new drug-like substances with improved cytotoxicity on lung cancer in humans.

A new depsidone of Lobaria pulmonaria with acetylcholinesterase inhibition activity.

The phytochemical investigation conducted on a foliose lichen, Lobaria pulmonaria(L.) Hoffm. (Lobariaceae), led to the isolation of a new depsidone in the form of its diacetate derivative which showed a moderate acetylcholinesterase inhibition activity (1 µg) in vitro. This is the first record of identified depsidone structure in searching for these inhibitors.

Cacospongionolide and scalaradial, two marine sesterterpenoids as potent apoptosis-inducing factors in human carcinoma cell lines.

Apoptosis, a form of programmed cell death, is a critical defence mechanism against the formation and progression of cancer and acts by eliminating potentially deleterious cells without causing such adverse effects, as inflammatory response and ensuing scar formation. Therefore, targeting apoptotic pathways becomes an intriguing strategy for the development of chemotherapeutic agents. In last decades, marine natural products, such as sesterterpenoids, have played an important role in the discovery and development of new drugs. Interestingly, many of these compounds have a strong potential as anticancer drugs by inhibiting cell proliferation and/or inducing cell death. In the present study, we investigated the effects of scalaradial and cacospongionolide, two sesterterpenoids from Cacospongia scalaris and Fasciospongia cavernosa marine sponges, on the apoptotic signalling pathway in three different human tumoral cells. Results were obtained by using DNA fragmentation, comet and viability assays, quantification of the mitochondrial transmembrane potential and Western blot. The T47D (human breast carcinoma), A431 (human epidermoid carcinoma), HeLa (human cervix carcinoma) and HCT116 (human colon carcinoma) cells were incubated for 24 h with scalaradial or cacospongionolide. Treatment of T47D cells with scalaradial or cacospongionolide for 24 h brought about a significant increase in DNA migration as well as fragmentation. Moreover, incubation of HCT116 and HeLa cells with scalaradial or cacospongionolide for 24 h caused an increased expression of pro-apoptotic proteins. Furthermore, scalaradial or cacospongionolide, added to HCT116 and HeLa cells overnight, induced a significant and concentration-dependent loss of mitochondrial transmembrane potential, an early apoptosis signalling event. These effects paralleled with those achieved with p50 and p65, NF-κB subunits, nuclear level. In conclusion, scalaradial and cacospongionolide, by determining human cancer cell apoptosis, may represent new promising compounds to inhibit cancer cell proliferation.

Acetylcholinesterase inhibition activity of acetylated depsidones from Lobaria pulmonaria.

As part of our ongoing project of new acetylcholinesterase inhibitors from lower marine and terrestrial species, a phytochemical investigation was conducted on a foliose lichen, Lobaria pulmonaria (L.) Hoffm. (Lobariaceae), from Bosnia and Herzegovina. The study led to the isolation of a mixture of acetylated depsidones which showed a moderate activity (0.5 µg) in the acetylcholinesterase inhibition test on Thin-layer chromatography plate. Our results indicate for the first time the significance of depsidones, highly specific metabolites from lichen species, in searching for these inhibitors which still represent the best drugs currently available for the management of Alzheimer's disease.