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... The cytotoxic effects of ZER were evaluated in cancer Caco-2 (undifferentiated), HeLa, and B16F10 cells by the MTT assay. Cancer cells, cultured as previously reported, 25,28 were incubated for 24 h with various concentrations of ZER (0.25 to 100 μM, dissolved in DMSO) in complete culture medium (treated cells). Treated cells were compared for viability to untreated cells (control cells, receiving no treatment) and vehicle-treated cells (incubated for 24 h with an equivalent volume of DMSO; maximal final concentration, 1%). ...
... At the end of the incubation time, cells were subjected to the MTT test as previously reported. 28 Preliminary evaluation of the morphologies of cancer cells after 24 h of incubation with various amounts of ZER was also performed by microscopic observation. Different concentrations of CUR (0.25 to 100 μM, from DMSO solution) were incubated for 24 h in B16F10 cells for cytotoxicity determination. ...
... Both undifferentiated Caco-2 cells, a line of human colon adenocarcinoma, and HeLa cells, a cell line derived from a human cervical epithelioid carcinoma, are widely used as models for oncological studies. 25,[28][29][30]39,40 Our observations of ZER cytotoxicity to cancer cells are in line with previous studies. ...
Article
The dietary sesquiterpene dienone zerumbone (ZER) targets selectively cancer cells inducing mitochondrial dysfunction and apoptosis, but protects non-cancerous cells towards oxidative stress and insult. This study examines the in vitro effect of ZER on lipid peroxidation in biological systems (cholesterol and phospholipid membrane oxidation) and explores its antitumor action in terms of its ability to modulate cancer cell lipid profile. Evaluation of the antioxidant activity of ZER showed that this compound is per se unable to trap lipoperoxyl radicals. ZER significantly modulated the total lipid and fatty acid profile in cancer cells, inducing marked changes in the phospholipid/cholesterol ratio, with a significant decrease in the level of oleic and palmitic acids and a marked increase of the stearic acid amount. Cell based fluorescent measurements of intracellular membranes and lipid droplets using the Nile Red staining technique showed that in cancer cells ZER induced a significant accumulation of cytosolic lipid droplets and altered cell membrane organization/protein dynamics, depolarizing the mitochondrial membranes and inducing apoptosis and alteration on nuclear morphology. The modulatory activity on total lipid and fatty acid profile and lipid droplets could therefore represent another possible mechanism for the anticancer properties of ZER.
... The cytotoxic effect of the nanoparticle formulation MO/ PF108 was evaluated in HeLa cells by the MTT assay (Rosa et al., 2013;Schiller et al., 1992). HeLa cells were seeded in 24-well plates at density of 3 Â 10 4 cells/well in 500 mL of serum-containing media. ...
... After treatment, the cells were scraped and centrifuged at 1200 g at 4 C for 5 min. After centrifugation, pellets were separated from supernatants and used for lipid extraction and analyses (Rosa et al., 2013). ...
... An aliquot of the CHCl 3 fraction from each cell sample was collected, dried down, and dissolved in 5 mL of EtOH. Separation of fatty acids was obtained by mild saponification (Rosa et al., 2013) as follows: 100 mL of Desferal solution (25 mg/mL of H 2 O), 1 mL of a water solution of ascorbic acid (25% w/v), and 0.5 mL of 10 N KOH were added to lipids in EtOH solution. The mixtures were left in the dark at room temperature for 14 h. ...
Article
Monoolein-based cubosomes are promising drug delivery nanocarriers for theranostic purposes. Nevertheless, a small amount of research has been undertaken to investigate the impact of these biocompatible nanoparticles on cell lipid profile. The purpose of the present investigation was to explore changes in lipid components occurring in human carcinoma HeLa cells when exposed to short-term treatments (2 and 4h) with monoolein-based cubosomes stabilized by Pluronic F108 (MO/PF108). A combination of TLC and reversed-phase HPLC with DAD and ELSD detection was performed to analyze cell total fatty acid profile and levels of phospholipids, free cholesterol, triacylglycerols, and cholesteryl esters. The treatments with MO/PF108 cubosomes, at non-cytotoxic concentration (83μg/mL of MO), affected HeLa fatty acid profile, and a significant increase in the level of oleic acid 18:1 n-9 was observed in treated cells after lipid component saponification. Nanoparticle uptake modulated HeLa cell lipid composition, inducing a remarkable incorporation of oleic acid in the phospholipid and triacylglycerol fractions, whereas no changes were observed in the cellular levels of free cholesterol and cholesteryl oleate. Moreover, cell-based fluorescent measurements of intracellular membranes and lipid droplet content were assessed on cubosome-treated cells with an alternative technique using Nile red staining. A significant increase in the amount of the intracellular membranes and mostly in the cytoplasmic lipid droplets was detected, confirming that monoolein-based cubosome treatment influences the synthesis of intracellular membranes and accumulation of lipid droplets. Copyright © 2015. Published by Elsevier Ireland Ltd.
... The cytotoxic effect of compounds was evaluated in cancer cells by the MTT assay (Schiller et al., 1992;Rosa et al., 2013). Cancer cells were seeded in 96-well plates at a density of 10 3 cells/mL (HeLa), 5 Â 10 4 cells/mL (Caco-2) and 3 Â 10 3 cells/mL (B16F10) in 100 mL of medium, and cultured overnight (Caco-2) or for 48 h (HeLa and B16F10). ...
... Cells were subsequently incubated for 24 h with various concentrations of Arz (Caco-2, HeLa, B16F10) and Me-Arz (Caco-2) in ethanol solution in complete culture medium; an equivalent volume of EtOH was added to the controls. After the cell medium removing and washing, cells were subjected to MTT test (Rosa et al., 2013). After incubation (4 h), color development was measured at 570 nm with an Infinite 200 auto microplate reader (Infinite 200, Tecan, Austria); the absorbance was proportional to the viable cell number. ...
... The influence of methylation was also investigated on cell viability. Arz and Me-Arz were compared for cytotoxicity in differentiated Caco-2 cell monolayers, an intestinal epithelial cell model extensively used in studies of toxicity, uptake, and metabolism of phenolic compounds (O'Brien et al., 2000;Tian et al., 2009;Rosa et al., 2011bRosa et al., , 2014, and in undifferentiated colonic epithelial cancer Caco-2 cells (Rosa et al., 2013). The structural modification of the lead compound didn't influence the effect on cell viability. ...
Article
The heterodimeric phloroglucinyl pyrone arzanol (Arz) has raised considerable interest because of its antiviral, anti-inflammatory, and antioxidant activity. We have investigated the effect of methylation of the pyrone moiety on the antioxidant activity and cytotoxicity of Arz. This manoeuvre, that left the polyphenolic moiety unscathed, was nevertheless detrimental for antioxidant activity in both the cholesterol thermal degradation- and the Cu²⁺-induced liposome oxidation assays, providing evidence of structure-activity relationships that go beyond the preservation of the polyphenolic pharmacophore. The antioxidant activity of Arz was retained also in the Fe-NTA model of in vivo oxidative stress, with protective effect on the oxidative degradation of plasmatic lipids, unsaturated fatty acids and cholesterol. Both Arz and methylarzanol (Me-Arz) were devoid of toxic effect on colonic differentiated Caco-2 cells up to 100 μM, but significantly reduced cancer Caco-2 cell viability at lower dosages. Arz could also selectively reduce viability of other cancer cell lines, [murine melanoma cells (B16F10 cells), human cervical carcinoma cells (HeLa cells)], suggesting that it can act as a selective modulator of cell processes typical of cancer cells. Taken together, our results qualify Arz as a lead structure for further in vivo investigation of its pharmacological potential.
... Bottarga is generally sold as whole ovaries (undamaged ovarian sacs) under vacuum packaging or grated (in jars or bags) [5,9]. We have previously studied the chemical composition [5,[8][9][10], stability to the oxidative degradation [9][10][11], non-enzymatic browning process [9][10][11], biological profile in normal and cancer cells [11][12][13], and bioavailability in cell systems and rat model [11][12][13][14] of Sardinian ...
... Bottarga is generally sold as whole ovaries (undamaged ovarian sacs) under vacuum packaging or grated (in jars or bags) [5,9]. We have previously studied the chemical composition [5,[8][9][10], stability to the oxidative degradation [9][10][11], non-enzymatic browning process [9][10][11], biological profile in normal and cancer cells [11][12][13], and bioavailability in cell systems and rat model [11][12][13][14] of Sardinian ...
... The chloroform/methanol/water (CHCl 3 /MeOH/H 2 O) mixture (in a ratio 2:1:1) was used for the extraction of total lipids from aliquots (40 mg) of grated bottarga samples (B1, B2 ad B3) as previously reported [9]. A dried aliquot of the CHCl 3 fractions from each bottarga sample was dissolved in methanol and injected into the high-performance liquid chromatograph (HPLC) system for the qualitative analysis of lipid components, while another dried aliquot, dissolved in ethanol, was subjected to mild saponification according to literature [9,12]. The unsaponifiable and saponifiable fractions were separated and total cholesterol (TC), total FA, and conjugated diene fatty acid hydroperoxides (HP) were analyzed by HPLC system [9,12]. ...
Article
Full-text available
A taste component is implicated in the oro-sensory detection of dietary lipids and free fatty acids seem to be involved in fatty food recognition. Bottarga, the salted and semi-dried ovary product of mullet (Mugil spp.), is a rich-fat food. A comparative sensory assessment of different commercial bottarga samples was performed in insect and human models in relation to their lipid composition. The bottarga attractant effect to Ceratitis capitata was assessed by behavioral tests. The subjective odor and taste perception of bottarga samples was investigated in human determining the rate of pleasantness, familiarity, and intensity dimensions using the 7-points Likert-type scale. Bottarga samples showed similar lipid profiles, but differences emerged in total and free fatty acid levels. Significant differences were observed in the attractant effect/acceptability of samples to medflies, negatively correlated to their total and free fatty acids. Insect female exhibited the ability to select among bottarga samples based on their visual and olfactory properties. In the human model, a potential contribution of free fatty acid amount in the pleasantness and familiarity dimensions of taste of bottarga samples was evidenced. Women exhibited a greater ability than men to select bottarga samples based on their better olfactory perception. Our results increase the knowledge about this outstanding product with nutritional and nutraceutical properties.
... The final product can be sold as whole ovaries under vacuum packaging or grated in jars. We have extensively studied the composition, 17,[22][23][24][25][26] oxidative stability, [23][24][25] browning process, 24,25 bioavailability 25,26 and biological profile 25,27,28 of bottarga samples manufactured in Sardinia. Bottarga represents an important natural, stable source of n-3 PUFA, 17,18,[22][23][24][25] in particular eicosapentaenoic acid (EPA, 20:5 n-3) and docosahexaenoic acid (DHA, 22:6 n-3), and a significant amount of these health beneficial fatty acids in mullet roe are in the form of wax esters. ...
... At the end of the incubation time, cells were subjected to the MTT test as previously reported. 27,28 Treated cells were compared for viability to untreated cells (control cells, receiving no treatment) and vehicle-treated cells (incubated for 24 h with an equivalent volume of EtOH; maximal final concentration, 1.2%). The cytotoxic effect of different concentrations (50-500 μg mL −1 , from a 40 mg mL −1 solution in EtOH) of BO, previously obtained from a grated bottarga, 28 was also tested in 3T3 cells for comparison. ...
... The total lipid content of mullet raw roes is estimated to be in the range 150-200 g kg −1 of fresh weight, 23 while the oil content of cured roes ranges from 230 to 325 g kg −1 of edible portion in whole bottarga 23 to 270-380 g kg −1 in grated bottarga samples. 17,[24][25][26][27] To date, several studies have been carried out for the extraction of oils from fish and fish waste using the SFE technique. 11,29,32 Supercritical CO 2 is an inert, non-toxic, environmentally safe solvent that allows the extraction at low temperatures/pressures and the obtained extracts are generally recognized as safe to be used in food product. ...
Article
BACKGROUND The Sardinian food delicacy “bottarga” is the final product of a number of treatments (salting and drying) on the ovaries of mullet (Mugil spp) and represents an important natural source of n‐3 polyunsaturated fatty acids (n‐3 PUFA) with nutraceutical properties. During the salting process of mullet roes to obtain bottarga, huge amounts of waste salt are generated, rich in residual ovary material. RESULTS We evaluated the lipid composition (main lipid components and fatty acids) and bioactivity of oil obtained from the ovary material separated from waste salt (waste salt oil). Oil was obtained by supercritical fluid extraction with CO2 (SFE‐CO2), an environmentally friendly separation technique. The lipid composition of waste salt oil was determined by ¹³C NMR spectroscopy and reversed‐phase HPLC‐DAD/ELSD chromatography. The oil was characterized by a relatively high level of n‐3 PUFA (122 ± 7 g kg‐1 of oil), and these beneficial health compounds were mainly present in the form of wax esters. Waste salt oil showed a marked cytotoxic effect (MTT assay) in cancer B16F10 melanoma cells, with a slight cytotoxic effect in normal cells (3T3 fibroblasts). Waste salt and its derivatives (salt oil and residual material after oil extraction) were also tested for the attractant effect and acceptability to insects (Ceratitis capitata ) to gain preliminary information about their potential application for animal supplementation. CONCLUSION The results qualify waste salt as a potential resource for veterinary dietary supplements, nutraceuticals, and pharmaceutical applications. This article is protected by copyright. All rights reserved.
... Among these, the salted and semi-dried mullet (Mugil genus) ovary product is a food delicacy produced in several world countries, with the name of bottarga in Italy, 14,15 avgotaracho in Greece, 16 and karasumi in Japan. 12 In recent works we have studied the lipid composition, 14,[17][18] oxidative stability, [18][19] browning processes, 20 water-soluble low molecular weight metabolite profile, 21,22 and biological activity 19,21 of bottarga (as whole and grated products) manufactured in Sardinia (Italy), that has a long tradition in making a high quality bottarga. The total lipid content of bottarga samples is estimated to be in the range 220-325 mg/g edible portion and its lipid fraction is characterized by a high portion of wax esters (ca. ...
... Among these, the salted and semi-dried mullet (Mugil genus) ovary product is a food delicacy produced in several world countries, with the name of bottarga in Italy, 14,15 avgotaracho in Greece, 16 and karasumi in Japan. 12 In recent works we have studied the lipid composition, 14,[17][18] oxidative stability, [18][19] browning processes, 20 water-soluble low molecular weight metabolite profile, 21,22 and biological activity 19,21 of bottarga (as whole and grated products) manufactured in Sardinia (Italy), that has a long tradition in making a high quality bottarga. The total lipid content of bottarga samples is estimated to be in the range 220-325 mg/g edible portion and its lipid fraction is characterized by a high portion of wax esters (ca. ...
... 14,18 Mullet bottarga represents an important natural, stable source of health beneficial EPA and DHA, that amount to 10-13 mg/g and 20-33.5 mg/g of edible portion, respectively (13-25% of have also shown the ability of bottarga lipids to reduce viability in colon adenocarcinoma cells and to induce a significant modification of the fatty acid composition in normal and cancer colon cells with a selective increase in n-3 PUFA levels, indicating the cellular bioavailability of these bioactive food components. 19,21 The bioavailability of n-3 PUFA may be influenced by the lipid structures in which they are incorporated (free fatty acids bound in ethyl esters, triacylglycerols, or phospholipids), and by the food matrix. 23,24 Several studies have reported the in vivo bioavailability (in humans or animal models) of n-3 PUFA from fish oils, algal oils, enriched foods, and fish roe oils. ...
Article
The effect of a diet enriched with mullet bottarga on the lipid profile (total lipids, total cholesterol, unsaturated fatty acids, α-tocopherol, and hydroperoxides) of plasma, liver, kidney, brain, and perirenal adipose tissues of healthy rats was investigated. Rats fed a 5 days 10% bottarga enriched-diet showed body weights and tissue total lipid and cholesterol levels similar to those of animals fed control diet. Univariate and multivariate results showed that bottarga enriched-diet modified the fatty acid profile in all tissues, except brain. Significant increases of n-3 PUFA, particularly EPA, were observed together with 20:4 n-6 decrease in plasma, liver, and kidney. Perirenal adipose tissue showed a fat accumulation that reflected diet composition. The overall data suggest that mullet bottarga may be considered a natural bioavailable source of n-3 PUFA and qualify it as a traditional food product with functional properties and potential functional ingredient for preparation of n-3 PUFA enriched foods.
... In our study, the attention was focused on the salted and dried mullet ovary product, a marine fat-rich food with nutritional and nutraceutical properties, produced in numerous world countries with different names, in Italy, is called "bottarga" (Scano et al., 2008), in Greece, "avgotaracho" (Kalogeropoulos, Nomikos, Chiou, Fragopoulou, & Antonopoulou, 2008), and in Japan, "karasumi" (Bledsoe, Bledsoe, & Rasco, 2003). Our previous studies analyzed bottarga chemical composition (Rosa et al., 2009(Rosa et al., , 2011Scano et al., 2008), stability to the oxidative degradation (Rosa et al., 2009(Rosa et al., , 2011, non-enzymatic browning process (Rosa et al., 2009(Rosa et al., , 2011, the effect on viability and lipid profile in normal and cancer cells (Rosa et al., 2011(Rosa et al., , 2016Rosa, Scano, Atzeri, Deiana, & Falchi, 2013), and bioavailability in cell systems and rat model (Rosa et al., 2011(Rosa et al., , 2013(Rosa et al., , 2016. Bottarga is considered a naturally rich source of n-3 polyunsaturated fatty acids (n-3 PUFA or ω-3 PUFA), such as eicosapentaenoic acid (EPA, 20:5 n-3) and docosahexaenoic acid (DHA, 22:6 n-3) with beneficial health effects (Rosa et al., 2009(Rosa et al., , 2016. ...
... In our study, the attention was focused on the salted and dried mullet ovary product, a marine fat-rich food with nutritional and nutraceutical properties, produced in numerous world countries with different names, in Italy, is called "bottarga" (Scano et al., 2008), in Greece, "avgotaracho" (Kalogeropoulos, Nomikos, Chiou, Fragopoulou, & Antonopoulou, 2008), and in Japan, "karasumi" (Bledsoe, Bledsoe, & Rasco, 2003). Our previous studies analyzed bottarga chemical composition (Rosa et al., 2009(Rosa et al., , 2011Scano et al., 2008), stability to the oxidative degradation (Rosa et al., 2009(Rosa et al., , 2011, non-enzymatic browning process (Rosa et al., 2009(Rosa et al., , 2011, the effect on viability and lipid profile in normal and cancer cells (Rosa et al., 2011(Rosa et al., , 2016Rosa, Scano, Atzeri, Deiana, & Falchi, 2013), and bioavailability in cell systems and rat model (Rosa et al., 2011(Rosa et al., , 2013(Rosa et al., , 2016. Bottarga is considered a naturally rich source of n-3 polyunsaturated fatty acids (n-3 PUFA or ω-3 PUFA), such as eicosapentaenoic acid (EPA, 20:5 n-3) and docosahexaenoic acid (DHA, 22:6 n-3) with beneficial health effects (Rosa et al., 2009(Rosa et al., , 2016. ...
Article
Full-text available
In this study, the role of sex and body weight in the olfactory and gustatory dimensions (pleasantness, intensity, and familiarity) of mullet cured roes, a marine rich-fat food with peculiar sensory attributes, was evaluated. One hundred seventy-seven participants were enrolled. Positive correlations were observed between all food taste and odor dimensions. Women reported a significantly higher odor and taste intensity ratings than men. Multivariate linear regression analyses evidenced that body weight in women was negatively correlated to the food odor and taste pleasantness and positively correlated to odor intensity. These negative correlations were due to different women gustatory performance in relation to body weight. A significantly lower perception of salty and bitter taste was observed in women with a body weight >60 kg compared to those with a body weight ≤60 kg. Our results underline the important role of sex and body weight in the food products sensory evaluation. Practical applications This study evidenced higher intensity ratings in women than men for the evaluation of olfactory and gustatory dimensions (pleasantness, intensity, and familiarity) of the salted and dried mullet roes, a lipid-rich food, and the role of body weight in women sensory perception. Therefore, our data highlight the importance of taking into consideration sex and body weight when consumers panels are selected and constituted for the evaluation of sensory properties and acceptance of lipid-rich foods, but also applicable to other types of foods.
... The leerfish Lichia amia is particularly known for its lipid profile, which is rich in monounsaturated and polyunsaturated fats, making it potentially important for cardiovascular health, but like Belone belone, it is a source of cholesterol, and therefore, it is necessary to consume the right amount to obtain positive health effects in this case as well [38]. Studies carried out on Mugil cephalus have paid particular attention to the importance of omega-3s in preventing colon cancer [119]. They are valid adjuvants of 5-fluorouracil, an anti-cancer drug [119]. ...
... Studies carried out on Mugil cephalus have paid particular attention to the importance of omega-3s in preventing colon cancer [119]. They are valid adjuvants of 5-fluorouracil, an anti-cancer drug [119]. Minimum levels of cholesterol are present in the surmullet Mullus surmulentus, which, on the other hand, shows high concentrations of vitamins of groups B and A but also of important minerals, such as zinc, which is important for processes involved in growth, tissue repair, and sexual development, and selenium, essential for its antioxidant and anti-cancer properties [38]. ...
Article
Full-text available
Lagoons play an important socio-economic role and represent a precious natural heritage at risk from fishing pressure and chemical and biological pollution. Our research focused on better understanding the discrimination of fish biodiversity, the detection of non-indigenous species, and the valorization of commercial indigenous species at Mellah lagoon (Algeria). Taxonomic characterization and barcoding for all fish species and Inkscape schematic drawings for the most common species are provided. A total of 20 families and 37 species were recorded. The thermophilic species Coris julis, Thalassoma pavo, and Aphanius fasciatus and tropical species such as Gambusia holbrooki and Parablennius pilicornis were identified. Numerous Mediterranean species of socio-economic importance are highlighted, and detailed information is summarized for the lagoon’s sustainability. This short-term evaluation goes hand in hand with long-term programs documenting the interaction between indigenous and non-indigenous species in the lagoon and will allow the development of a provisional relationship model for future studies. Thermophilic and tropical species patterns in the Mellah lagoon are presented. Taken together, we provide useful data that can guide future investigations and may become a potential management tool for achieving the Sustainable Development Goals and protecting species with large socio-economic roles from potential thermal stress impact.
... The leerfish Lichia amia is particularly known for its lipid profile, which is rich in monounsaturated and polyunsaturated fats, making it potentially important for cardiovascular health, but like Belone belone, it is a source of cholesterol, and therefore, it is necessary to consume the right amount to obtain positive health effects in this case as well [38]. Studies carried out on Mugil cephalus have paid particular attention to the importance of omega-3s in preventing colon cancer [119]. They are valid adjuvants of 5-fluorouracil, an anti-cancer drug [119]. ...
... Studies carried out on Mugil cephalus have paid particular attention to the importance of omega-3s in preventing colon cancer [119]. They are valid adjuvants of 5-fluorouracil, an anti-cancer drug [119]. Minimum levels of cholesterol are present in the surmullet Mullus surmulentus, which, on the other hand, shows high concentrations of vitamins of groups B and A but also of important minerals, such as zinc, which is important for processes involved in growth, tissue repair, and sexual development, and selenium, essential for its antioxidant and anti-cancer properties [38]. ...
Poster
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Aquatic resources are geographically defined but they are not always protected in their environment, cultural and economic value. In this context, the present overview aims to throws light on the biota background of the Algerian Basin El Mellah with a focus on its alieutic source which transit from the Mediterranean Sea. As reported in many studies, there is an alarming fall of marine organism catches, and this leads to a research activity about the aulietic species identification for a correct monitoring and development of the basin. Since 2016, basin biodiversity has been studied with a morphological approach and on an evolutionary scale (nucleotide sequences analysis) in order to report (i) the taxonomic category of benthic and pelagic macrofauna; (ii) the traceability of gene product in GenBank (ww.ncbi.com); (iii) the potential of barcoding (mathematical approach to taxonomy support); (iv) the degree of species vulnerability (ww.iucn.com); (v) the discrimination of endemic and species of commercial interest; (vi) the construction of species phylogenetic relations. Five phylum have been identified, as well as Echinoderms (2 species), Polychaete (14 species), Molluscs (14 species), Crustaceans (10 species) and Fish (38 species). Three organisms found are into endangered species category such as Gammarus aequicauda, Anguilla anguilla, and Hippocampus ramulosu. Further separation based on sequence discrimination showed missing barcoding percentage ([n° of species found with unknown sequence / n° of total species found]*100) showed 50% for Echinoderms, 64.3% for Polychaete and Mollusc respectively, 60% for Crustaceans, 15.8% for total Fish founded and 47.4% only for commercial Fish. Thus, it is necessary to provide training approaches and theoretical methods for a correct species identification in order to increase missing barcoding percentage with a useful marker for discriminate phylogenetic relationships and above all the reproductive species health, which is a necessary condition for the their sustainability and the ecosystem equilibrium of El Mellah Basin.
... They reported that the n-3 PUFA showed a high stability and very minor oxidation during processing and storage, indicating that mullet roe may be regarded as stable natural source of n-3 PUFA and, therefore, beneficial for human health. Other results showed interesting anti-tumour properties of bottarga lipids, and qualify this fish product as a food with nutraceutical properties and potential benefits in colon cancer prevention (Rosa et al., 2013). ...
Article
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This short review compiles the data concerning the quality of flesh, processed roe and smoked fillets of grey mullets (Mugilidae), including when available the data concerning the fish from freshwater and in particular those from Tunisian reservoirs. The biochemical and mineral composition of the fish, the nutritional benefits and the potential health risks related to the consumption of fish products are discussed with regard to human health, taking into consideration the nutritional recommendations and normative toxicological limits defined by leading health authorities. Flesh proximal compositions of fish from freshwater are relatively scarce, but the few available lipid data are within the very large proximal range (from less than 1% to more than 11%) reported for grey mullets in general. Most of the studies reveal the predominance of unsaturated fatty acids with a substantial proportion v3 type not only for fish from marine environment but also for those from freshwater. Flesh constitutes a source of essential amino acids and mineral nutrients too. The mullets are generally reported to be safe for human consumption except the fish from heavily contaminated zones. Globally, it should be regarded as an interesting contribution to a healthy diet. The mullets are also greatly appreciated for the production of salted and dried roe as well as smoked fillets, which allow increasing their economic value while preserving products health beneficial attributes. Data concerning processing mullet's roe from freshwater are missing, but it may be supposed that these roes have acceptable texture, taste and flavour as they are highly sought by connoisseurs. It is recommended to perform studies on the qualities of freshwater mullet's roe and to look for labelling the origin for both the roes and the smoked fillets of mullet from Tunisian reservoirs dedicated to the production of drinking water as it would guarantee that the fish come from unpolluted environments.
... Biogenic amines, caused by amino acids degradation, were determined on commercial dried roes (Kung et al., 2008) also during a 180 day storage period (Restuccia et al., 2015). The Mugil cephalus roe oil was found to have functional properties, being a potential bioavailable source of omega-3 PUFA (Rosa, Atzeri, Putzu, & Scano, 2016) with potential benefits in cancer prevention Rosa, Scano, Atzeri, Deiana, & Falchi, 2013). ...
Article
The aim of this work was to measure the physico-chemical and the colorimetric parameters of ovaries from Mugil cephalus caught in the Tortolì lagoon (South-East coast of Sardinia) along the steps of the manufacturing process of Bottarga, together with the rheological parameters of the final product. A lowering of all CIELab coordinates (lightness, redness and yellowness) was observed during the manufacture process. All CIELab parameters were used to build a Linear Discriminant Analysis (LDA) predictive model able to determine in real time if the roes had been subdued to a freezing process, with a success in prediction of 100%. This model could be used to identify the origin of the roes, since only the imported ones are frozen. The major changes of all the studied parameters (p < 0.05) were noted in the drying step rather than in the salting step. After processing, Bottarga was characterized by a pH value of 5.46 (CV=2.8) and a moisture content of 25% (CV=8), whereas the typical per cent amounts of proteins, fat and NaCl, calculated as a percentage on the dried weight, were 56 (CV=2), 34 (CV=3) and 3.6 (CV=17), respectively. The physical chemical changes of the roes during the manufacturing process were consistent for moisture, which decreased by 28%, whereas the protein and the fat contents on the dried weight got respectively lower of 3% and 2%. NaCl content increased by 3.1%. Principal Component Analyses (PCA) were also performed on all data to establish trends and relationships among all parameters. Hardness and consistency of Bottarga were negatively correlated with the moisture content (r=−0.87 and r=−0.88, respectively), while its adhesiveness was negatively correlated with the fat content (r=−0.68).
... In a preliminary study, we showed that oil obtained from MM collected in Sardinia induced cytotoxicity on tumor cells (cancer Caco-2) with no cytotoxic action on normal cells (differentiated Caco-2 cells) under the conditions employed [12]. In this study we compared the effect of MM fixed oil on cell viability and lipid composition in two different cancer cell lines: the undifferentiated Caco-2 cells, a line of human colon adenocarcinoma, amply used for oncological studies investigating the anticarcinogenic effects of food constituents [28][29][30], and the B16F10 murine melanoma cells, a highly metastatic cancer cell line abundantly used to screen antitumor natural substances and extracts [17,[19][20][21]31]. ...
Article
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The present study aimed to examine the potential anticancer properties of fixed oil obtained from Maltese mushroom (Cynomorium coccineum L.), an edible, non-photosynthetic plant, used in traditional medicine of Mediterranean countries to treat various ailments and as an emergency food during the famine. We investigated the effect of the oil, obtained from dried stems by supercritical fractioned extraction with CO2, on B16F10 melanoma and colon cancer Caco-2 cell viability and lipid profile. The oil, rich in essential fatty acids (18:3n-3 and 18:2n-6), showed a significant growth inhibitory effect on melanoma and colon cancer cells. The incubation (24 h) with non-toxic oil concentrations (25 and 50 μg/mL) induced in both cancer cell lines a significant accumulation of the fatty acids 18:3n-3 and 18:2n-6 and an increase of the cellular levels of eicosapentaenoic acid (20:5n-3) with anticancer activity. Moreover, the oil exhibited the ability to potentiate the growth inhibitory effect of the antitumor drug 5-fluorouracil in Caco-2 cells and to influence the melanin content in B16F10 cells. The results qualify C. coccineum as a resource of oil, with potential benefits in cancer prevention, for nutraceutical and pharmaceutical applications.
... Epidemiological studies have shown that populations consuming large amounts of LC n-3 PUFA-rich fish oil have a lower risk of CRC [14]. In vitro studies have found that EPA and DHA exert their effects on cancer cells via several mechanisms including changing the membrane composition, altering intracellular Ca ++ concentrations as well as intracellular pH, modifying mitochondrial membrane potential/permeability, changing cellular resistance to ROS damage, and by direct actions on DNA and gene expression [15][16][17][18][19][20][21][22][23]. Animal studies have also shown that fish oil supplementation reduced the number and size of tumours, angiogenesis and metastasis [24][25][26][27][28]. ...
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Background: Currently available treatments for colorectal cancer (CRC) associate with numerous side-effects that reduce patients' quality of life. The effective nutraceuticals with high anti-proliferative efficacy and low side-effects are desirable. Our previous study has reported that free fatty acids extract (FFAE) of krill oil induced apoptosis of CRC cells, possibly associated with changes in mitochondrial membrane potential (MMP). The aims of this study were to compare the anti-proliferative efficacy of FFAE from krill oil on CRC cells with commonly used chemotherapeutic drug, Oxaliplatin, and to investigate the molecular mechanisms underlying the anti-proliferative effects of krill oil with a focus on intrinsic mitochondrial death pathway. Methods: Three human CRC cell lines, including DLD-1, HT-29 and LIM-2405, and one mouse CRC cell line, CT-26, were treated with FFAE of KO and the bioactive components of krill oil, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) for 24 h and 48 h. Similarly, these cell lines were treated with Oxaliplatin, a commonly used drug for CRC treatment, for 24 h. The effects of FFAE of KO, EPA, DHA and Oxaliplatin on cell proliferation, mitochondrial membrane potential and reactive oxygen species (ROS) were determined via WST-1, JC-10, and ROS assays respectively. The expression of caspase-3, caspase-9 and DNA damage following treatments of FFAE of KO was investigated via western blotting and immunohistochemistry. Results: The FFAE of KO, EPA and DHA significantly inhibited cell proliferation and increased formation of ROS in all four cell lines (P < 0.01). A small dose of FFAE from KO ranged from 0.06 μL/100 μL to 0.12 μL/100 μL containing low concentrations of EPA (0.13-0.52 μM) and DHA (0.06-0.26 μM) achieved similar anti-proliferative effect as Oxaliplatin (P > 0.05). Treatments with the FFAE of KO, EPA and DHA (2:1 ratio) resulted in a significant increase in the mitochondrial membrane potential (P < 0.001). Furthermore, the expression of active forms of caspase-3 and caspase-9 was significantly increased following the treatment of FFAE of KO. Conclusions: The present study has demonstrated that the anti-proliferative effects of krill oil on CRC cells are comparable with that of Oxaliplatin, and its anti-proliferative property is associated with the activation of caspase 3/9 in the CRC cells.
... The cytotoxic effects of methanol extracts, alone or in the presence of TBH, were preliminarily evaluated in Caco-2 cells by the MTT assay (Rosa, Scano, Atzeri, Deiana, & Falchi, 2013). Caco-2 cells were seeded, at a passage number of 75-77, in 96-well plates at a density of 5 × 10 5 cells/mL in 100 μL of medium and cultured at confluence. ...
... Compared to tumor-bearing mice alone, treatment with Cis (40 mg/Kg) injection, HD-H post-EAC-cells inoculation increased the number of splenocytes significantly, and the number of splenocytes in EAC-bearing mice treated with Cis (40 mg/Kg) was higher than this number in mice bearing tumor and treated with HD-H (100 µg/Kg). The present data werein agreement with the previous studies, which demonstrated increased splenocytes count in tumor-bearing mice treated with chemotherapeutic agents (Merritt et al., 2003;Park et al., 2009;Rosa et al., 2013;Zhu et al., 2019;Hashem et al., 2020). ...
... The cytotoxic effects of methanol extracts, alone or in the presence of TBH, were preliminarily evaluated in Caco-2 cells by the MTT assay (Rosa, Scano, Atzeri, Deiana, & Falchi, 2013). Caco-2 cells were seeded, at a passage number of 75-77, in 96-well plates at a density of 5 × 10 5 cells/mL in 100 μL of medium and cultured at confluence. ...
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... Cells were cultured in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal calf serum (FCS), 2 mM L-glutamine, and penicillin (100 units/mL)-streptomycin (100 µg/mL), at 37 °C in 5% CO 2 . The cytotoxic effect of saffron extracts was evaluated in cancer Caco-2 cells by the MTT assay (Rosa, Scano, Atzeri, Deiana, & Falchi, 2013;Schiller, Klainz, Mynett, & Gescher, 1992). Caco-2 cells were seeded in 96-well plates at a density of 5 × 10 4 cells/mL in 100 µL of medium and cultured overnight. ...
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Fish roe is a prized food that has been recognized to have substantial value since ancient times, especially in the Mediterranean region. Roes from mullet, catfish, and carp are appreciated in Middle Eastern and African markets due to their availability, nutritional and sensory properties. Adding value to roe and extending shelf life has been directed through methods of processing, preservation, and achieving sensory attributes desired by consumers. For example, there has been increasing interest in mullet Bottarga all over the world as a promising food in terms of nutritional value, nutraceutical properties and potential health benefits. Appropriate procedures for processing and storage have been the necessary strategy to preserve the safety and quality of this healthy food. Typically, eviscerated mullet roe is salted, pressed and dried under controlled humidity and temperature. Many biochemical changes are observed during the salting and drying process, and as roe products are susceptible to browning due to oxidation, appropriate manufacturing conditions are required. Processed roe, such as mullet Bottarga represents an important natural health source of long-chain omega-3 polyunsaturated fatty acids (n-3PUFA), in particular eicosapentaenoic acid (EPA, 20:5n-3) and docosahexaenoic acid (DHA, 22:6n-3), which are reported to have health benefiting potential. Further fish roes commonly reported discussed.
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(1)H NMR spectroscopy was employed to study the modifications over time of the water-soluble low molecular weight metabolites extracted from samples of salted and dried mullet (Mugil cephalus) roes (mullet bottarga) stored at different conditions. Samples of grated mullet bottarga were stored for 7 months at -20 °C, at 3 °C, and at room temperature in the presence and in the absence of light and then timely extracted and analyzed by NMR. Principal component multivariate data analysis applied to the spectral data indicated that samples stored at -20 °C maintained similar features over time whereas, along PC1, samples stored at room temperature in the presence and in the absence of light showed, over time, marked metabolite modifications. The comparative analysis of the integrated areas of the selected regions of the (1)H NMR spectra indicated that the major compositional changes due to storage conditions were (i) the increase of the derivatives of the breakdown of phosphatidylcholine (choline, phosphorylcholine, and glycerol), (ii) the breakdown of nucleosides, (iii) the decrease of methionine, tryptophan, and tyrosine, and (iv) the cyclization of creatine. These changes were observed at different storage conditions, with more pronounced trends in the samples stored at room temperature. The role of metabolites in food aging is discussed.
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Unlabelled: The salted and semidried mullet (Mugil cephalus) ovary product (bottarga) is proposed as an important source of the long-chain n-3 polyunsaturated fatty acids (PUFA), eicosapentaenoic acid, and docosahexaenoic acid. In this work, we investigated the extent of lipid oxidation and browning of grated bottarga samples during 7 mo of storage at -20 °C, 2 to 3 °C in the absence of light, and at room temperature in the presence or absence of light. Modifications of the levels of total choline (as index of phospholipid breakdown), total sugars, and free amino acids such as lysine, methionine, and tryptophan (involved in nonenzymatic browning) were also studied at different storage conditions. Storage of bottarga did not significantly affect the n-3 PUFA and cholesterol levels with respect to the control; nevertheless, a significant hydroperoxide increase was observed during 7 mo in bottarga samples at all the storage conditions, while low malondialdeyde levels were measured. Samples placed at room temperature in the absence and in the presence of light showed over time a marked browning process, lipid breakdown, a sensible decrease in the levels of total sugars, tryptophan, and methionine with respect to control and samples stored at -20 °C and 2 to 3 °C. The resistance against the oxidation of the isolated bottarga lipids was also assessed in dry state at several temperatures (37, 75, and 140 °C). Practical application: We evaluated the change in lipid compounds and color of dried and salted mullet roes under different storage conditions. The obtained results suggest the importance of the low temperatures to preserve the nutritional properties of this fish product during long storage.
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Nuclear magnetic resonance (NMR) spectroscopy combined with multivariate data analysis (MVA) was used to investigate the molecular components of the aqueous extract of samples of bottarga, that is, salted and dried mullet (Mugil cephalus) roe, manufactured in Sardinia (Italy) from mullets of known and unknown geographical provenience. Principal component analysis (PCA) applied to the processed (1)H NMR spectra indicated that samples tend to cluster according to their geographical origin and also on the basis of storage and manufacturing procedures. The most important metabolites that characterized grouping of samples are the free amino acids methionine (Met), glutamate (Glu), histidine (His), phenylalanine (Phe), tyrosine (Tyr), and isoleucine (Ile); trimethylamine (TMA) and dimethylamine (DMA), both biomarkers of degradation; nucleotides and derivatives; choline (Cho) and phosphorylcholine (P-cho); and lactate (Lac).
Article
The importance of n-3 polyunsaturated fatty acid (n-3 PUFA) intake has long been recognized in human nutrition. Although health benefits, n-3 PUFA are subject to rapid and/or extensive oxidation during processing and storage, resulting in potential alteration in nutritional composition and quality of food. Bottarga, a salted and semi-dried mullet ( Mugil cephalus ) ovary product, is proposed as an important source of n-3 PUFA, having high levels of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). In this work, we investigated the extent of lipid oxidation of grated bottarga samples during 7 months of storage at -20 °C and room temperature under light exposure. Cell viability, lipid composition, and lipid peroxidation were measured in intestinal differentiated Caco-2 cell monolayers after 6-48 h of incubation with lipid and hydrophilic extracts obtained from bottarga samples at different storage conditions. The storage of bottarga did not affect the n-3 PUFA level, but differences were observed in hydroperoxide levels in samples from different storage conditions. All tested bottarga extracts did not show a toxic effect on cell viability of differentiated Caco-2 cells. Epithelial cells incubated with bottarga oil had significant changes in fatty acid composition but not in cholesterol levels with an accumulation of EPA, DHA, and 22:5. Cell hydroperoxides were higher in treated cells, in relation to the oxidative status of bottarga oil. Moreover, the bottarga lipid extract showed an in vitro inhibitory effect on the growth of a colon cancer cell line (undifferentiated Caco-2 cells).
Article
The viability and state of proliferation of cells in culture is conveniently assessed using MTT, which is metabolically reduced to stain functionally intact cells. The hypothesis was tested that this assay can also be used in the quantitation of effects of toxicants on hepatocytes in suspension. Hepatocytes isolated from BALB/c mice were incubated without or with menadione, rotenone, N-methylformamide or paracetamol. Cellular damage was measured by either MTT assay or release into the medium of lactate dehydrogenase (LDH). Results obtained with the two tests were compatible in the case of toxicity inflicted by menadione or N-methylformamide. Rotenone decreased cell viability as indicated by the MTT assay immediately after addition of the agent, whereas measurement of LDH release did not detect this rapid toxic effect. The MTT assay detected paracetamol-induced damage within the first hour of exposure, but this was not detected by the LDH assay until 3 hr had elapsed.
Article
An in vitro study showed that a lipid emulsion containing fish oil (FO) slows the growth of colon cancer cells and enhances their sensitivity to 5-fluorouracil (FU). The aim was to confirm this finding and to compare such an emulsion with an alternative to lowered n-6 fatty acid exposure. We determined the number of viable cells, apoptosis and cell cycle distribution of HT-29 cells after exposure to one of three lipid emulsions. Cell cycle distribution was also assessed after treatment with lipid emulsions and FU. The lipid emulsion containing FO induced a significant growth inhibitory effect without changing the percentage of apoptotic cells. Exposure to the other lipid emulsions had no effect on growth and decreased apoptosis. Each lipid emulsion potentiated the S phase-halting effect of 1 and 10microM FU. This effect also occurred at 0.1microM FU when the cells were exposed to the FO containing lipid emulsion. A lipid emulsion containing FO has a growth inhibitory effect on a human colon adenocarcinoma cell line, an effect not due to the induction of apoptosis, and potentiated the S phase-halting effect of FU. Thus, an FO lipid emulsion may be of benefit in colorectal cancer.
Article
Attention to the role of n-3 long-chain fatty acids in human health and disease has been continuously increased during recent decades. Many clinical and epidemiologic studies have shown positive roles for n-3 fatty acids in infant development; cancer; cardiovascular diseases; and more recently, in various mental illnesses, including depression, attention-deficit hyperactivity disorder, and dementia. These fatty acids are known to have pleiotropic effects, including effects against inflammation, platelet aggregation, hypertension, and hyperlipidemia. These beneficial effects may be mediated through several distinct mechanisms, including alterations in cell membrane composition and function, gene expression, or eicosanoid production. A number of authorities have recently recommended increases in intakes of n-3 fatty acids by the general population. To comply with this recommendation a variety of food products, most notably eggs, yogurt, milk, and spreads have been enriched with these fatty acids. Ongoing research will further determine the tissue distribution, biological effects, cost-effectiveness, and consumer acceptability of such enriched products. Furthermore, additional controlled clinical trials are needed to document whether long-term consumption or supplementation with eicosapentaenoic acid/docosahexaenoic acid or the plant-derived counterpart (alpha-linolenic acid) results in better quality of life.
Article
We report that the dye nile red, 9-diethylamino-5H-benzo[alpha]phenoxazine-5-one, is an excellent vital stain for the detection of intracellular lipid droplets by fluorescence microscopy and flow cytofluorometry. The specificity of the dye for lipid droplets was assessed on cultured aortic smooth muscle cells and on cultured peritoneal macrophages that were incubated with acetylated low density lipoprotein to induce cytoplasmic lipid overloading. Better selectivity for cytoplasmic lipid droplets was obtained when the cells were viewed for yellow-gold fluorescence (excitation, 450-500 nm; emission, greater than 528 nm) rather than red fluorescence (excitation, 515-560 nm; emission, greater than 590 nm). Nile red-stained, lipid droplet-filled macrophages exhibited greater fluorescence intensity than did nile red-stained control macrophages, and the two cell populations could be differentiated and analyzed by flow cytofluorometry. Such analyses could be performed with either yellow-gold or red fluorescence, but when few lipid droplets per cell were present, the yellow-gold fluorescence was more discriminating. Nile red exhibits properties of a near-ideal lysochrome. It is strongly fluorescent, but only in the presence of a hydrophobic environment. The dye is very soluble in the lipids it is intended to show, and it does not interact with any tissue constituent except by solution. Nile red can be applied to cells in an aqueous medium, and it does not dissolve the lipids it is supposed to reveal.
Article
The popularity of epithelial cell lines in studies of drug transport processes can probably be explained by the ease with which new information is derived from these rather simple in vitro models. Drug transport studies in epithelial cell monolayers grown on permeable supports are easy to perform under controlled conditions, resulting in the generation of a wealth of information. This information can generally be relatively easily translated into fundamental principles, since the homogenous epithelial cell cultures lack the complexity and variability found in more complex whole tissue models. In fact, much of our more recent knowledge about active and passive drug transport mechanisms has been obtained from studies in various epithelial cell cultures (1). The good correlation between passive drug transport through various epithelial cell monolayers and that seen across the human intestine in vivo now makes it possible to use the cell cultures for studies on structure-absorption relationships (examples are found in refs. 2-4). Indeed, both cell culture and transport experiments across epithelial cell monolayers have been automated, and these models are now used as a screening tool in drug discovery programs in many drug companies for the prediction of intestinal drug permeability.
Article
Fish roe products are extremely valuable and currently enjoy expanding international and domestic markets. Caviars represent the best-known form of fish roe products; however, several other product forms are also consumed, including whole skeins and formulations with oils and cheese bases. Caviars are made from fish roe after the eggs have been graded, sorted, singled-out, salted or brined, and cured. Most caviar is marketed as a refrigerated or frozen food. Several types of caviar from different fish species are marketed as shelf-stable products. Market preferences for lower salt content have raised food safety concerns. Descriptions of and processing technologies for many delightful fish roe and caviar food products are presented here.
Article
Epidemiological studies suggest that polyunsaturated fatty acids may protect against colorectal neoplasia. In order to explore this observation, cell proliferation and viability, lipid composition, membrane fluidity, and lipid peroxidation were measured in Caco-2 cells after 48h incubation with various fatty acids. Saturated and monounsaturated fatty acids incorporated less well in the membranes than polyunsaturated fatty acids (PUFAs). All of the PUFAs tested had an inhibitory effect on cell proliferation/viability whereas the saturated and monounsaturated fatty acids did not. Addition of palmitic acid had no significant effect on membrane fluidity whereas unsaturated fatty acids increased membrane fluidity in a dose-dependent manner. PUFAs strongly increased tumor cell lipid peroxidation in a dose-dependent manner. Saturated and monounsaturated fatty acids increased lipid peroxidation in this cell line only at high concentration. Preincubation of Caco-2 cells with vitamin E prevented the inhibition of proliferation/viability, the elevation of the MDA concentration and the increased membrane fluidity induced by PUFAs. Our data indicate that PUFAs are potent inhibitors of the growth of colon cancer cells in vitro.
Article
The incidence of colon cancer in industrialised countries has increased since the early 1970s. It is estimated that more than one-third of cases are associated with factors related to a Western diet. Both the type and amount of dietary fats consumed have been implicated in colon cancer aetiology. Recent studies have demonstrated that n-3 polyunsaturated fatty acids (PUFAs), commonly found in fish oil (FO), could prevent colon cancer development. Evidences show that n-3 PUFAs act at different stages of cancer development and through several mechanisms including the modulation of arachidonic acid-derived prostaglandin synthesis, and Ras protein and protein kinase C expression and activity. As a result, n-3 PUFAs limit tumour cell proliferation, increase apoptotic potential along the crypt axis, promote cell differentiation and possibly limit angiogenesis. The modulatory actions of n-3 PUFAs on the immune system and their anti-inflammatory effects might also play a role in reducing colon carcinogenesis. There remains, nevertheless, some ambiguity over the safety of n-3 PUFAs with respect to secondary tumour formation. However, it appears that n-3 PUFAs may be of use in colon cancer prevention.
Article
N-6 polyunsaturated fatty acids (PUFAs) may be associated with increased risk of colon cancer, whereas n-3 PUFAs may have a protective effect. We examined the effects of docosahexaenoic acid (DHA), eicosapentaenoic acid and arachidonic acid on the colon carcinoma cell lines SW480 derived from a primary tumour, and SW620 derived from a metastasis of the same tumour. DHA had the strongest growth-inhibitory effect on both cell lines. SW620 was relatively more growth-inhibited than SW480, but SW620 also had the highest growth rate in the absence of PUFAs. Flow cytometry revealed an increase in the fraction of cells in the G2/M phase of the cell cycle, particularly for SW620 cells. Growth inhibition was apparently not caused by increased lipid peroxidation, reduced glutathione or low activity of glutathione peroxidase. Transmission electron microscopy revealed formation of cytoplasmic lipid droplets after DHA treatment. In SW620 cells an eightfold increase in total cholesteryl esters and a 190-fold increase in DHA-containing cholesteryl esters were observed after DHA treatment. In contrast, SW480 cells accumulated DHA-enriched triglycerides. Arachidonic acid accumulated in a similar manner, whereas the nontoxic oleic acid was mainly incorporated in triglycerides in both cell lines. Interestingly, nuclear sterol regulatory element-binding protein 1 (nSREBP1), recently associated with cell growth regulation, was downregulated after DHA treatment in both cell lines. Our results demonstrate cell-specific mechanisms for the processing and storage of cytotoxic PUFAs in closely related cell lines, and suggest downregulation of nSREBP1 as a possible contributor to the growth inhibitory effect of DHA.
Article
Polyunsaturated free fatty acids (PUFAs) participate in normal functioning of the cell, particularly in control intracellular cell signalling. As nutritional components they compose a human diet with an indirect promoting influence on tumourogenesis. The PUFAs level depends on the functional state of the membrane. This work is focused on changes only of free unsaturated fatty acids amount (AA - arachidonic acid, LA - linoleic acid, ALA - alpha-linolenic acid, palmitoleic acid (PA) and oleic acid) in cell membranes of colorectal cancer of pT3 stage, G2 grade without metastasis. Qualitative and quantitative composition of free unsaturated fatty acids in the membrane was determined by high-performance liquid chromatography. It was shown that the malignant transformation was accompanied by a decrease in amount of LA and ALA while arachidonic and oleic acids increased. It is of interest that free AA levels are elevated in colon cancer, as AA is the precursor to biologically active eicosanoids.
Article
Polyunsaturated fatty acid (PUFA) rich micro-algal oil was tested in vitro and compared with fish oil for antiproliferative properties on cancer cells in vitro. Oils derived from Crypthecodinium cohnii, Schizochytrium sp. and Nitzschia laevis, three commercial algal oil capsules, and menhaden fish oil were used in cell viability and proliferation tests with human colon adenocarcinoma Caco-2 cells. With these tests no difference was found between algal oil and fish oil. The nonhydrolysed algal oils and fish oil showed a much lower toxic effect on cell viability, and cell proliferation in Caco-2 cells than the hydrolysed oils and the free fatty acids (FFAs). Eicosapentaenoic acid (EPA; C20:5n-3) and docosahexaenoic acid (DHA; C22:6n-3) were used as samples for comparison with the tested hydrolysed and nonhydrolysed oils. The hydrolysed samples showed comparative toxicity as the free fatty acids and no difference between algal and fish oil. Oxidative stress was shown to play a role in the antiproliferative properties of EPA and DHA, as alpha-tocopherol could partially reverse the EPA/DHA-induced effects. The results of the present study support a similar mode of action of algal oil and fish oil on cancer cells in vitro, in spite of their different PUFA content.
Article
In this study we examined the effects of a fish oil-based lipid emulsion (FO) rich in omega-3 fatty acids, which is used in humans as a component of parenteral nutrition, on the growth of the colon cancer cell line Caco-2. The aim of the present study was to investigate whether the FO influences growth and chemosensitivity of the colon cancer cell line Caco-2. FO was tested alone and in combination with the anticancer drug 5-fluorouracil (5-FU). Cell numbers were determined with crystal violet staining, cell cycle distribution was assessed using a flow cytometer and apoptosis was visualized by staining nuclei with diamino-phenylindole hydrochloride. FO inhibited growth of Caco-2 cells in a time and dose dependent manner. FO treatment evoked apoptosis as confirmed by cell morphology. Cell cycle analysis identified an accumulation of cells in the G(2)/M phase after incubation with FO. The combined treatment of the cells with FO and 5-FU resulted in a significant enhancement of the growth inhibition seen after exposure to either substance alone. Treatment of the cells with 5-FU specifically blocked the cell cycle in the S phase. The combined treatment of 5-FU with FO showed a further increase in the accumulation of cells in the S phase. In conclusion, FO has a potent antiproliferative effect on Caco-2 cells, at least in part, due to a decrease in the progression of the cell cycle and the induction of apoptosis. The combination of FO with 5-FU results in an additive growth inhibitory effect.
Article
A growing body of epidemiological, clinical, and experimental evidence has underscored both the pharmacological potential and the nutritional value of dietary fish oil enriched in very long chain n-3 PUFAs such as docosahexaenoic acid (DHA, 22:6, n-3) and eicosapentaenoic acid (EPA, 20:5, n-3). The broad health benefits of very long chain n-3 PUFAs and the pleiotropic effects of dietary fish oil and DHA have been proposed to involve alterations in membrane structure and function, eicosanoid metabolism, gene expression and the formation of lipid peroxidation products, although a comprehensive understanding of the mechanisms of action has yet to be elucidated. In this review, we present data demonstrating that DHA selectively modulates the subcellular localization of lipidated signaling proteins depending on their transport pathway, which may be universally applied to other lipidated protein trafficking. An interesting possibility raised by the current observations is that lipidated proteins may exhibit different subcellular distribution profiles in various tissues, which contain a distinct membrane lipid composition. In addition, the current findings clearly indicate that subcellular localization of proteins with a certain trafficking pathway can be subjected to selective regulation by dietary manipulation. This form of regulated plasma membrane targeting of a select subset of upstream signaling proteins may provide cells with the flexibility to coordinate the arrangement of signaling translators on the cell surface. Ultimately, this may allow organ systems such as the colon to optimally decode, respond, and adapt to the vagaries of an ever-changing extracellular environment.
Article
Crude composition, lipid composition, and tocopherols, ascorbic acid, cholesterol, phytosterols, and squalene content together with fatty acids and antiplatelet activities of total, neutral, and polar lipids of avgotaracho (wax-covered, dried, and salted Mugil cephalus roe) were studied and compared with those of similar products. Wax and steryl esters accounted for 63.7% of roe lipids followed by phosphatidylcholine (PC), which comprised 20.3%. Wax esters were rich in saturated fatty alcohols, monounsaturated fatty acids, and long chain omega3 highly unsaturated fatty acids (HUFA). The fatty acid distribution in roe total and neutral lipids was similar to that of wax esters, while in polar lipids, the omega3 HUFA predominated. Avgotaracho provides significant amounts of protein, fat, alpha-tocopherol, ascorbic acid, and PC, certain amounts of squalene and phytosterols, and cholesterol at levels comparable to hens' eggs. Total, polar, and neutral lipids of avgotaracho exhibited a strong inhibition of platelet activating factors and thrombin, with polar lipids being more active. The results obtained indicate that avgotaracho is a food of high nutritive value, rich in protein and lipids with a healthy lipid profile in terms of omega3/omega6 ratio and major fatty acid classes, while the antiplatelet activity of its oil indicates a putative antithrombotic potential.
Article
Dietary intake of long-chain omega-3 (or n-3) polyunsaturated fatty acids (PUFA), eicosapentaenoic acid (EPA), and docosahexaenoic acid (DHA) can affect numerous processes in the body, including cardiovascular, neurological and immune functions, as well as cancer. Studies on human cancer cell lines, animal models and preliminary trials with human subjects suggest that administration of EPA and DHA, found naturally in our diet in fatty fish, can alter toxicities and/or activity of many drugs used to treat cancer. Multiple mechanisms are proposed to explain how n-3 PUFA modulate the tumor cell response to chemotherapeutic drugs. n-3 PUFA are readily incorporated into cell membranes and lipid rafts, and their incorporation may affect membrane-associated signaling proteins such as Ras, Akt and Her-2/neu. Due to their high susceptibility to oxidation, it has also been proposed that n-3 PUFA may cause irreversible tumor cell damage through increased lipid peroxidation. n-3 PUFA may increase tumor cell susceptibility to apoptosis by altering expression or function of apoptotic proteins, or by modulating activity of survival-related transcription factors such as nuclear factor-kappaB. Some studies suggest n-3 PUFA may increase drug uptake or even enhance drug activation (e.g., in the case of some nucleoside analogue drugs). Further research is warranted to identify specific mechanisms by which n-3 PUFA increase chemotherapy efficacy and to determine the optimal cellular/membrane levels of n-3 PUFA required to promote these mechanisms, such that these fatty acids may be prescribed as adjuvants to chemotherapy.
Decreased polyunsaturated fatty acid content contributes to increased survival in human colon cancer A systemic review of the roles of n-3 fatty acids in health and disease
  • M Oraldi
  • A Trombetta
  • F Biasi
  • R A Canuto
  • M Maggiora
  • G Muzio
Oraldi, M., Trombetta, A., Biasi, F., Canuto, R.A., Maggiora, M., Muzio, G., 2009. Decreased polyunsaturated fatty acid content contributes to increased survival in human colon cancer. J. Oncol., 867915. Riediger, N.D., Othman, R.A., Suh, M., Moghadasian, M.H., 2009. A systemic review of the roles of n-3 fatty acids in health and disease. J. Am. Diet. Assoc. 109, 668– 679.
Taurine as drug and functionl food component Glutathione and Sulfur Amino Acids in Human Health and Disease
  • R C Gupta
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  • M Masella
Gupta, R.C., D'Archivio, M., Masella, R., 2009. Taurine as drug and functionl food component. In: Masella, R., Mazza, G. (Eds.), Glutathione and Sulfur Amino Acids in Human Health and Disease. John Wiley & Sons, Hoboken, NJ, USA, pp. 519–541.
Glutathione and Sulfur Amino Acids in Human Health and Disease
  • R C Gupta
  • M D'archivio
  • R Masella
Gupta, R.C., D'Archivio, M., Masella, R., 2009. Taurine as drug and functionl food component. In: Masella, R., Mazza, G. (Eds.), Glutathione and Sulfur Amino Acids in Human Health and Disease. John Wiley & Sons, Hoboken, NJ, USA, pp. 519-541.
Application of epithelial cell culture in studies of drug transport
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  • J Gråsjö
  • J Taipalensuu
  • G Ocklind
  • P Artursson
Travelin, S., Gråsjö, J., Taipalensuu, J., Ocklind, G., Artursson, P., 2002. Application of epithelial cell culture in studies of drug transport. In: Wise, C. (Ed.), Epithelial Cell Culture Protocols. Humana Press, Totowa, NJ, pp. 233-272.
A comparative study on the effect of algal and fish oil on viability and cell proliferation of Caco-2 cells
  • V A Van Beelen
  • J Roeleveld
  • H Mooibroek
  • L Sijtsma
  • R J Bino
  • D Bosh
  • I M C M Rietjens
  • G M Alink
Van Beelen, V.A., Roeleveld, J., Mooibroek, H., Sijtsma, L., Bino, R.J., Bosh, D., Rietjens, I.M.C.M., Alink, G.M., 2007. A comparative study on the effect of algal and fish oil on viability and cell proliferation of Caco-2 cells. Food Chem. Toxicol. 45, 716-724.