Fatty Acid Ratios as Parameters to Discriminate Between Normal and Tumoral Cells and Compare Drug Treatments in Cancer Cells

The fatty acid (FA) composition of cell membranes represents a metabolic biomarker. However, the FA profile reproducibility in cell cultures remains a significant challenge. In this study, cell FA ratios are validated as metabolic markers alternative to cell FA. To this goal, cell samples belonging to cancer HeLa cells and normal 3T3 fibroblasts, from various experimental sets, are analyzed by a high-performance liquid chromatography system coupled with a photodiode array detector and evaporative light scattering detector (HPLC-DAD/ELSD), and the ratios among the main FA are calculated. Principal component analysis (PCA) separately performed on FA and FA ratio data indicates similar clustering of cell samples concerning the cell type. Moreover, similar scores values t[1] and t[2] and graphical distances are calculated in the PCA plots separately performed on FA and FA ratios measured in cancer HeLa cells subjected to various antitumoral compounds. Last, PCA applied to selected FA ratios measured in various cell lines, obtained in similar experimental conditions, allows to discriminate between normal and tumoral cells. The results substantiate FA ratios as a cell-specific fingerprint, characterized by reproducibility across intra-laboratory conditions, useful for cell characterization, discrimination between normal and tumoral cells, and the comparison of different drug treatments. Practical Applications: The reproducibility of the fatty acid (FA) profile in cell cultures remains a significant challenge. Results obtained from this study improve knowledge about the role of the FA ratio profile as a cell-specific fingerprint characterized by reproducibility across intra-laboratory conditions. The characterization of the specific FA ratio profile of a cell culture, under standardized experimental conditions, can facilitate the comparative evaluation of cell data sets for nutritional, metabolic, and pharmacological studies, overcoming differences in cell culture conditions and FA extraction/analytical procedures

The Role of Lipid Composition in the Sensory Attributes and Acceptability of the Salted and Dried Mullet Roes (Bottarga): A Study in Human and Animal Models

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

Cytogenetic identity: A new parameter for estimating whole-genome differences

In this paper, we introduce a novel parameter, called Cytogenetic Identity, to describe differences and similarities between genomes. Using Whole Comparative Genomic Hybridization plus Digital Image Analysis, we present a new methodology that employs the whole genome, including highly repeated DNA sequences, to provide a general picture about the differences between individuals of the same or different species. The proposed approach has a great potential in many different fields of research, like evolution, ecology, phylogenesis, etc. In the present study, we applied Cytogenetic Identity to establish a quantitative degree of divergence between different goat breeds. Advantages as well as disadvantages of the new parameter are discussedThe work was supported by Fondo Integrativo per la Ricerca (F.I.R.), provided by the University of Cagliari (to M.N. and R.R.

Antimicrobial, antioxidant and anti-tyrosinase properties of extracts of the Mediterranean parasitic plant Cytinus hypocistis

Background: Cytinus is an endophytic parasitic plant occurring in South Africa, Madagascar, and in the Mediterranean region. We have extracted the inflorescences (the only visible part of the plant, emerging from the host roots at the time of blossom) of Cytinus hypocistis collected in Sardinia, Italy, and explored the antimicrobial, antioxidant, anti-tyrosinase, and cytotoxic activities of the extracts. Methods: Extracts from C. hypocistis were prepared using increasing polarity solvents: cyclohexane, ethanol, and water. Phenolic composition were determined through spectrophotometric assays, and antioxidant activity with both electron-transfer and hydrogen-atom assays. Nine different bacterial strains, including clinical isolate methicillin-resistant Staphylococcus aureus, were used in agar diffusion method. Cytotoxicity was tested using against the B16F10 melanoma cell line. Results: While cyclohexane extracts where biologically inactive, ethanolic and aqueous extracts displayed an intriguing activity against several Gram-positive bacterial strains, including methicillin-resistant S. aureus, and against the Gram-negative Acinetobacter baumanii. Compared to the conventional antibiotics like cloxacillin, ampicillin, and oxytetracycline, C. hypocistis extracts were less active in absolute terms, but displayed a wider spectrum (notably, cloxacillin and ampicillin were inactive against methicillin-resistant S. aureus). The ethanolic extract of C. hypocistis was found to be particularly rich in polyphenols, in most part hydrolysable tannins. The antioxidant activity of extracts, tested with several methodologies, resulted to be particularly high in the case of ethanolic extracts, in accordance with the composition in phenolics. In detail, ethanol extracts presented about a twofold higher activity than the water sample when tested through the oxygen radical absorbance capacity-pyrogallol red (ORAC-PYR) assay. Cytotoxicity analysis against the B16F10 melanoma cell line showed that both extracts have not significant cytotoxic effect, even at the highest dose (1000 μg/mL). Tests showed that ethanolic extracts also had the greatest tyrosinase inhibition activity, indicating that C. hypocistis-derived substances could find application in food formulations as anti-browning agents. Conclusions: Overall, these results point to the need of further studies on C. hypocistis extracts, aimed at isolating and fully characterizing its biologically active compounds. © 2015 Zucca et al

Maltese mushroom (Cynomorium coccineum L.) as source of oil with potential anticancer activity

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 mu g/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

Evolution of satellite DNA sequences in two tribes of Bovidae: a cautionary tale

Two clones, Bt1 from Bos taurus and Om1 from Ovis orientalis musimon, were used as probes for hybridization on genomic DNA and on metaphase chromosomes in members of Bovini and Caprini tribes. Bt1 and Om1 are sequences respectively belonging to the 1.715 and 1.714 DNA satellite I families. Southern blots and fluorescence in situ hybridization experiments showed completely coherent results: the Bovini probe Bt1 hybridized only to members of the Bovini tribe and not to members of Caprini. Likewise, the Caprini probe Om1 hybridized only to members of the Caprini tribe and not to members of Bovini. Hybridization signals were detected in the heterochromatic regions of every acrocentric autosome, except for two pairs of autosomes from Capra hircus that did not show hybridization to probe Om1. No signal was detected on X and Y chromosomes or on bi-armed autosomes. Remarkably, probe Om1 showed almost 100% homology with a bacterial sequence reported in Helicobacter pylor

Evaluation of the Antioxidant and Cytotoxic Activities on Cancer Cell Line of Extracts of Parasitic Plants Harvested in Tunisia

Weeds thrive in agricultural environments. However, in certain areas of the world, they are consumed by humans as food, and they can represent a source of valuable active ingredients of ethnomedical interest. In this study, three North African parasitic plants of the Orobanchaceae family, Cistanche violacea, Orobanche crenata and Orobanche lavandulacea, normally considered as weeds, were studied to compare their biological activities. The antioxidant activity of hexane, ethyl acetate, acetone, methanol, and water extracts obtained by sequential maceration was estimated by DPPH, ABTS and FRAP assays. The metabolic profi les of aqueous extracts were studied by 1H NMR spectroscopy. The oils extracted using hexane as solvent were analyzed by GC-FID and HPLC-DAD to characterize the fatty acid profi le. The cytotoxic effect of the oil from the parasitic plants was also studied on B16F10 melanoma cancer cells. All extracts showed antioxidant activity and differences between extracts obtained using different solvents as well as for different plants have been highlighted. The highest total phenolic content was determined in O. crenata acetone extract (3.53±0.20 mmol GAE/g) and in O. lavandulacea methanol extract (2.29±0.02 mmol GAE/g). 1H NMR spectroscopy revealed the presence of amino acids, organic acids and carbohydrates. The major fatty acids in the oils were 18:2 n-6, 16:0, 18:1 n-9, and 18:3 n-3. O. lavandulacea revealed to be a highly cytotoxic species for B16F10 melanoma cells. These data suggest that, despite the extreme climate conditions and the damage that can result, the studied desert parasitic plants can represent a novel and precious source of metabolites, with promising biological activities

Biological activities and nutraceutical potentials of water extracts from different parts of Cynomorium coccineum L. (Maltese Mushroom)

Maltese Mushroom (Cynomorium coccineum L.) is a non-photosynthetic plant that has been used in traditional medicine for many centuries. In this paper, water extracts from the whole plant, external layer and peeled plant were studied to determine the main components responsible for its biological activities, i.e., its antimicrobial, antioxidant, and anti-tyrosinase activities; its cytotoxicity against mouse melanoma B16F10 cells; and its pro-erectile activity in adult male rats. The results of electron transfer and hydrogen transfer assays showed that the antioxidant activity was mainly due to anthocyanins in the external layer, whereas the external layer and peeled plant extracts both inhibited the microbial growth of several Gram-positive strains. In contrast, the whole plant extract had the highest anti-tyrosinase activity and exhibited pro-erectile activity when administered subcutane-ously. Overall, this study elucidated which parts of Maltese Mushroom are responsible for its antimicrobial, antioxidant, and anti-tyrosinase activities and thus which extracts have potential for use in nutraceutical formulation

Cholinium-Based Ionic Liquids from Hydroxycinnamic Acids as New Promising Bioactive Agents: A Combined Experimental and Theoretical Investigation

Cholinium-based ionic liquids (Cho-ILs) are very attractive compounds for medicinal and pharmaceutical applications as bioavailability enhancers in drug formulations and active components in pharmaceutical ingredients. In this study, we synthesized six Cho-ILs from hydroxycinnamic acids (HCA) ([Cho][HCA] ILs), a group of bioactive compounds with poor water solubility. [Cho][HCA] ILs and their parent acids were evaluated for solubility, thermal stability, and antioxidant activity. Furthermore, [Cho][HCA] ILs were screened for their cytotoxicity. To rationalize the experimental antioxidant activities, density functional theory (DFT) calculations were performed. The theoretical approach allowed for identification of the most likely radical scavenging mechanisms involving HCAs and the corresponding ionic forms under the studied experimental conditions and to rationalize the observed activity differences between salts and acids. Overall, our results revealed a higher solubility and free-radical scavenging activity for [Cho][HCA] ILs than corresponding HCAs, a relatively high thermal stability (melting temperature > 100 °C) and negligible cytotoxicity activity. Furthermore, DFT calculations showed that both the hydrogen atom transfer and the sequential proton loss electron transfer mechanisms are likely to occur in aqueous and ethanolic solutions. The picture emerging from our results supports the increasingly expressed idea that [Cho][HCA]ILs are promising candidates for applications in pharmaceutical formulatio