Investigating isotopic markers for hazelnut geographical authentication: Promising variables and potential applications.

Hazelnuts' features and price are influenced by their geographical origin, making them susceptible to fraud, especially counterfeit claims regarding their provenance. Stable isotope analysis is a recognised approach to establish the geographical origin of foods, yet its potential in hazelnut authentication remains unexplored. In this prospective study, we assessed multiple isotopic markers in hazelnuts from different origins and evaluated the most promising variables for geographical authentication by chemometric tools. Our findings indicate that bulk δ18O, along with δ2H and δ13C in the main fatty acid methyl esters, exhibit significant potential in discriminating geographical origins, and 87Sr/86Sr analysis could serve as a proficient confirmatory tool. Though no single marker alone can differentiate between all the studied origins, employing a multi-isotopic approach based on PLS-DA models achieved up to 92.5 % accuracy in leave-10 %-out cross-validation. These findings will probably lay the groundwork for developing robust models for hazelnut geographical authentication based on larger datasets.

Prospective exploration of hazelnut's unsaponifiable fraction for geographical and varietal authentication: A comparative study of advanced fingerprinting and untargeted profiling techniques.

This study compares two data processing techniques (fingerprinting and untargeted profiling) to authenticate hazelnut cultivar and provenance based on its unsaponifiable fraction by GC-MS. PLS-DA classification models were developed on a selected sample set (n = 176). As test cases, cultivar models were developed for "Tonda di Giffoni" vs other cultivars, whereas provenance models were developed for three origins (Chile, Italy or Spain). Both fingerprinting and untargeted profiling successfully classified hazelnuts by cultivar or provenance, revealing the potential of the unsaponifiable fraction. External validation provided over 90 % correct classification, with fingerprinting slightly outperforming. Analysing PLS-DA models' regression coefficients and tentatively identifying compounds corresponding to highly relevant variables showed consistent agreement in key discriminant compounds across both approaches. However, fingerprinting in selected ion mode extracted slightly more information from chromatographic data, including minor discriminant species. Conversely, untargeted profiling acquired in full scan mode, provided pure spectra, facilitating chemical interpretability.

Effect of feeding olive pomace acid oil on pork lipid composition, oxidative stability, colour, and sensory acceptance.

One of the targets of the meat industry is to reduce production costs and to increase the sustainability of the food chain, which has driven the attention towards the use of by-products as feed ingredients. Acid oils are fat by-products coming from the chemical refining process of edible oils, with a high energy value and that are approved as feed ingredients in the European Union. However, meat producers are hesitant to utilise them due to their varying composition and the limited understanding of their impact on animal performance and meat quality. The objective of this study was to evaluate the effects of using olive pomace acid oil (OPAO) instead of its corresponding crude olive pomace oil (OPO) or crude palm oil (PO) in pig diets on lipid composition, lipid oxidation and quality of pork loin (longissimus dorsi), fresh and after commercial refrigerated storage for 8 days. The experimental design consisted of feeding pigs with four diets supplemented with a 5% of PO, OPO, OPAO or a blend (B) of PO and OPAO (50:50, w/w). Fresh and refrigerated pork loin samples were assessed for fatty acid profile; tocopherol (T) and tocotrienol (T3) composition; lipid oxidative stability with the ferrous oxidation-xylenol orange method; 2-thiobarbituric acid (TBA) value; volatile compounds; colour; and sensory acceptance. Results showed that refrigeration reduced the total T + T3 levels and increased the TBA values and the volatile compound concentrations. The refrigerated storage also affected the instrumental colour parameters (L*, a* and b*) but not the overall acceptance of pork. Regarding the diet, pork from OPAO diet showed a higher unsaturated-to-saturated fatty acid ratio than pork from PO and B diets. The lowest T + T3 concentration was found in OPO and OPAO fresh pork and in OPAO refrigerated pork. The oxidative stability of fresh pork was lower for OPAO than for PO diet, but no significant effect of the diet was observed for this parameter in refrigerated pork. The TBA values and volatile compound concentrations of fresh pork were not affected by the diet. After refrigeration, OPAO pork had the highest TBA value and volatile compound concentrations. In any case, colour and consumer acceptance of pork were not affected by diet. In conclusion, in order to upcycle acid oils in pig diets, and considering results on the lipid oxidative stability of pork, it would be preferable to add the OPAO used in this study blended with PO.

Influence of free fatty acid content and degree of fat saturation in laying hen diets on egg quality, yolk fatty acid profile, and cholesterol content.

The aim of the present study was to evaluate the effect of dietary free fatty acid (FFA) content and the degree of saturation on egg quality, yolk fatty acid (FA) profile, and yolk cholesterol content. For a 15-wk period, a total of 144 laying hens (19-wk-old) were randomly assigned to 8 treatments arranged in a 2 × 4 factorial design, with 2 sources of crude oil (soybean oil and palm oil) and 4 levels of FFA (10, 20, 30, and 45%). The dietary treatments were achieved by progressively substituting the original oils with equivalent amounts of their corresponding acid oils (soybean acid oil and palm fatty acid distillate, respectively). No differences in ADFI or egg mass were found. However, dietary FFA reduced egg production (linear, P < 0.05) and increased the feed conversion ratio (linear, P < 0.05). Higher levels of FFA in soybean diets resulted in higher egg weight with higher albumen and yolk weights (linear, P < 0.01). Palm diets presented higher yolk:albumen ratio than soybean diets (P < 0.001), but the effect of FFA did not follow a linear trend. Hens fed soybean diets laid eggs with higher Haugh units (HU) than palm diets (P < 0.001), although increasing the dietary FFA% reduced the HU values in both (linear, P < 0.001). Palm diets enhanced shell quality with greater resistance to breakage, and higher dry matter and ash content than soybean diets (P < 0.05). No differences in egg chemical composition and yolk cholesterol content were found (P > 0.05). The saturation degree had a significant effect on all the analyzed yolk FA (P < 0.001) except for arachidonic acid (C20:4 n-6), whereas increasing the FFA content did not affect to a great extent. These results show that varying dietary FFA level did not affect egg quality and yolk composition as much as the dietary fat source did, supporting the use of acid oils and fatty acid distillates as fat ingredients for feed.

Olive pomace oil and acid oil as alternative fat sources in growing-finishing broiler chicken diets.

The aim of the present study was to investigate the effect of dietary supplementation of olive pomace oil and olive pomace acid oil, which are rich in monounsaturated fatty acids (FA) but differ in free FA content, on growth performance, digestibility and FA profile of abdominal fat and breast meat. A total of 3,048 one-day-old mixed-sex broiler chickens (Ross 308) were randomly distributed into 24 pens and 3 dietary treatments (8 replicates per treatment). Experimental diets were administered for growing (from 22 to 29 d) and finishing (from 30 to 39 d) periods, consisting of a basal diet supplemented with 6% (as-fed basis) palm oil (PO), olive pomace oil (O), or olive pomace acid oil (OA). Animals fed O achieved the lowest feed conversion ratio (P < 0.01), together with the highest AME value (P = 0.003), but no differences were observed between OA and PO. Regarding FA digestibility, O and OA showed higher values than PO for all FA in both apparent ileal digestibility (AID) and apparent total tract digestibility. Comparing the AID between O and OA, no differences were observed for total FA, monounsaturated FA, or polyunsaturated FA, but animals fed OA showed lower AID values for saturated FA than those fed O (P < 0.001). The FA profile of abdominal fat and breast meat reflected that of the diet, with higher monounsaturated FA and lower saturated FA in animals fed O and OA compared to those fed PO. In sum, the inclusion of both olive pomace oil and acid oil in growing-finishing broiler chicken diets led to great performance parameters and high FA digestibility values, together with an enrichment with monounsaturated FA in abdominal fat and breast meat compared to the use of palm oil. However, a better AID of saturated FA and feed conversion ratio is achieved with O compared to OA.

Effects of free-fatty-acid content and saturation degree of the dietary oil sources on lipid-class content and fatty-acid digestibility along the gastrointestinal tract in broilers from 22 to 37 days of age.

The aim of the present study is to assess the effect of the free-fatty-acid (FFA) content and saturation degree of dietary fat (added at 6%) on the fatty-acid (FA) digestibility and lipid-class content along the gastrointestinal tract and excreta in broilers from 22 to 37 d of age. This is essential to determine the potential use of acid oils (refining by-products rich in FFA) in broiler diets as an alternative to crude oils. The study consisted of a 2 × 4 factorial arrangement, which included 2 fat sources (soybean oils - unsaturated, or palm oils - saturated) and 4 levels of FFA (5, 15, 35, and 50%). Samples of digestive content of the gizzard, duodenum, jejunum, ileum and of the excreta were obtained at 37 d of age. Irrespective of the dietary fat source, more than 80% of total FA (TFA) was absorbed in the jejunum. Broilers fed with unsaturated diets had a higher absorption efficiency of FA than did those fed with saturated diets. This conclusion is supported by the lower FFA content and the higher TFA and polyunsaturated FA (PUFA) digestibility coefficients in the ileum (P < 0.001) observed in the former group. The dietary FFA level did not affect the FA absorption process as much as the dietary fat source did. This was supported by the lack of statistical differences among the diets with a similar saturation degree but rather different levels of FFA, for TFA, saturated FA, and PUFA digestibility coefficients both in the jejunum and ileum. However, the interactions reported in the ileum for triacylglycerol and diacylglycerol contents (P < 0.001), as well as for monounsaturated FA digestibility coefficients (P < 0.05) show that the dietary FFA content affects the FA absorption process. The present results show that the inclusion of acid oils in grower-finisher broiler diets with FFA levels up to 35% does not have a negative impact on the FA absorption process.

Effects of dietary free fatty-acid content and saturation degree on lipid-class composition and fatty-acid digestibility along the gastrointestinal tract in broiler starter chickens.

The aim of the present study is to assess the effect of the dietary free fatty acid (FFA) content and dietary fat saturation degree on the fatty-acid (FA) digestibility and lipid-class content along the gastrointestinal tract and excreta in broiler chickens. The 8 experimental diets resulted from replacing crude soybean oil with soybean acid oil from chemical refining, or crude palm oil with palm FA distillate from physical refining. Thus, there were 4 soybean and 4 palm diets with 6% added fat varying in their FFA% (5, 15, 35, and 50%). Samples of digestive content (gizzard, duodenum, jejunum, and ileum) and excreta were collected at 14 D for the determination of the FA digestibility and lipid-class content. The total FA digestibility coefficients reported for the chickens fed S diets in the jejunum, ileum, and excreta were higher than for those fed P diets (P ≤ 0.02). The general greater digestibility of the unsaturated diets was mainly explained by a higher contribution of the ileum to the absorption of saturated FA. The dietary FFA content mainly affected the FA absorption process. The diets with 50% FFA presented lower saturated FA digestibility coefficients in the jejunum and ileum (P ≤ 0.03), and higher content of FFA in the ileum and excreta (P ≤ 0.014), in comparison to the diets with 5% FFA. The 15% FFA diets were not different from the 5% FFA diets, regarding the saturated FA digestibility in the jejunum and excreta, and the FFA content in the ileum and excreta. It was concluded that unsaturated diets with moderate content of dietary FFA (up to 15%) could be used in broiler-chicken starter diets, as they led to similar FA absorption and performance results to the diets with the lowest dietary FFA content. From the present study, it has also been concluded that dietary saturated FA content has a greater impact on FA absorption than the dietary FFA content has.

Buckypaper-Bilirubin Oxidase Biointerface for Electrocatalytic Applications: Buckypaper Thickness.

Electrode materials play an important role on the electrocatalytic properties of immobilized biocatalysts. In this regard, achieving direct electronic communication between the electrode and redox sites of biocatalysts eliminates the need for additional electron transfer mediators for biocatalytic applications in fuel cells and other electrochemical energy devices. In order to increase electrocatalytic currents and power in fuel cells and metal-air batteries, conductive carbon-nanostructure-modified large surface area electrodes are quite useful. Among various electrode materials, freestanding buckypapers made from carbon nanotubes have gained significance as they do not require a solid support material and thus facilitate miniaturization. In this article, we present the effect of buckypaper (BP) thickness on the electrocatalytic properties of a bilirubin oxidase (BOD) enzyme. In this study, we prepared BPs of varying thicknesses ranging from 87 µm, the minimum thickness for suitable handling with a good stability in aqueous experiments, to 380 µm. BOD was adsorbed overnight onto the BPs, mostly via hydrophobic and π-π interactions since the nanotubes used were not chemically functionalized. Furthermore, intercalation of the BOD molecules onto the nanotubes' multicylindrical network is feasible. We determined that the lower range BP thickness (<220 µm) exhibited better sigmoidal shaped electrocatalytic currents than the higher BP-thickness-based BOD biofilms with larger capacitive currents. An oxygen reduction current density of up to 3 mA cm-2 is achieved without the use of any redox mediators or tedious electrode modifications. Using the 87 µm thick BP as the representative case, we were able to obtain distinguishable peaks for all Cu sites of BOD and assign their types, T1, T2, and T3, based on the peak-width at half-maximum in anaerobic cyclic voltammograms. Our peak assignment is further supported by the appearance of dual electrocatalytic oxygen reduction waves at a higher scan rate region (>10 mV s-1) in oxygen-saturated buffer, which is identified to be driven by an ∼3.5 times faster electron transfer rate from the buckypaper to the T2/T3 center than the T1 Cu site. Findings from this study are significant for designing enzyme electrocatalytic systems and biosensors in general and fuel cells and aerobic energy storage devices in particular, where the cathodic oxygen reduction current is often inadequate.

Evolution of lipid classes and fatty acid digestibility along the gastrointestinal tract of broiler chickens fed different fat sources at different ages.

The aim of the present study is to evaluate the effect of the dietary fat saturation degree and age on the lipid class (TAG, DAG, MAG, and FFA) composition and fatty acid digestibility along the gastrointestinal tract (GIT) and excreta in broiler chickens. A total of 120 one-day-old female broiler chickens were randomly distributed in 2 dietary treatments (6 cages/treatment), which resulted from the supplementation of a basal diet with 6% of soybean oil or palm oil. Two digestibility balances were carried out at 14 and 35 d and fatty acid digestibility and lipid class composition were determined in the gizzard, duodenum, jejunum, ileum, and excreta. Along de GIT, both fatty acid digestibility and lipid class composition were influenced by the dietary fat source and the age of the chickens. The absorption of the unsaturated fat was more efficient and faster than it was for the saturated fat. The ability of adult chickens to absorb fat was higher than for young chickens. The results show that the duodenum is the main place of fat digestion (hydrolysis), and the jejunum the main place of fat absorption. The role of the ileum on fat absorption is very important, as it is the last segment of the GIT where the absorption of fatty acids has been described. Thus, it was the contribution of the ileum that was responsible for the higher fat utilization observed for animals fed the unsaturated diet than for those fed the saturated diet at 14 d, and it was also responsible for the improvement on the utilization of the saturated diet between 14 and 35 d. All the results suggest that the absorption of fatty acids is more limiting than is hydrolysis, because the main differences were observed in the jejunum and ileum, where the absorption of fatty acids takes place.

The different molecular structure and glycerol-to-fatty acid ratio of palm oils affect their nutritive value in broiler chicken diets.

The aim of this study is to assess how the fat molecular structure and its glycerol-to-fatty acid ratio (G : FA) affect the fatty acid (FA) apparent absorption of palm oils in broiler chickens. The experimental diets were the result of a basal diet supplemented with 6% of different palm oils. Native palm oil (N), rich in triacylglycerols, was the positive control (T1), and acid palm oil (A), rich in free FA, was the negative control (T2). In order to improve the nutritive value of A, two different nutritional strategies were performed. The first strategy was achieved by adding increasing amounts of free glycerol (G) (4% (T3), 8% (T4) and 16% (T5)) to A, and the second one by adding increasing amounts of mono- (MAG) and diacylglycerols (DAG), coming from re-esterified palm oil (E) (40% (T6), 70% (T7), and 100% (T8)) to A. As a result, eight dietary treatments were formulated with a G : FA ratio ranging from 0.04 to 0.67. These treatments were randomly assigned to 192 one-day-old female broiler chickens (Ross 308), distributed in 48 cages. The results showed how, by keeping the G : FA ratio constant (0.33 mol/mol), the diet with a high MAG and DAG content (T7) achieved higher saturated FA apparent absorption values than did the diet with a high triacylglycerol content (T1) and this, in turn, more than did the diet with a high free FA content (T4). The behavior of oils with high or low G : FA ratio was dependent on whether G was in a free state or esterified as part of acylglycerol molecules. Thus, increasing amounts of G to A did not enhance the total FA apparent absorption, but rather quite the opposite, even impairing the absorption of mono- and polyunsaturated FA. However, increasing amounts of E (rich in MAG and DAG) to A (rich in FFA) did enhance total FA apparent absorption, primarily due to the increased absorption of saturated FA. In conclusion, the greater the G : FA ratio of a palm oil, the greater the absorption of total FA, as long as G is esterified as part of acylglycerol molecules. Thus, the re-esterification process for obtaining E makes sense in order to give added value to A, achieving even greater digestibility values than does its corresponding N.

Use of re-esterified palm oils, differing in their acylglycerol structure, in fattening pig diets.

Re-esterified oils are new fat sources obtained from the chemical esterification of acid oils with glycerol (both economically interesting by-products from oil refining and biodiesel industries, respectively). The different fatty acid (FA) positional distribution and acylglycerol composition of re-esterified oils may enhance the apparent absorption of saturated fatty acids (SFA) and, therefore, their overall nutritive value, which might lead to an increased deposition of SFA. The aim of the present study was to investigate the potential use of re-esterified palm oils, in comparison with their corresponding acid and native oils in fattening pig diets, studying their effects on fatty acid apparent absorption, acylglycerol and free fatty acid (FFA) composition of feces, growth performance, carcass-fat depots and fatty acid composition of backfat. Seventy-two crossbred boars and gilts (average weight of 24.7 ± 2.55 kg) were blocked by initial BW (nine blocks of BW for each gender), housed in adjacent individual boxes, and fed one of the four dietary treatments, which were the result of a basal diet supplemented with 4% (as-fed basis) of native palm oil (PN), acid palm oil (PA), re-esterified palm oil low in mono- and diacylglycerols (PEL), or re-esterified palm oil high in mono- and diacylglycerols (PEH). Regarding results from the digestibility balance, PA and PN showed similar apparent absorption coefficients (P>0.05), despite the high, FFA content of the former. However, re-esterified palm oils (both PEL and PEH) showed a higher apparent absorption of total FA than did their corresponding native and acid oils (P0.05). We conclude that re-esterified oils are interesting fat sources to be considered in fattening pigs.

Use of re-esterified palm oils, differing in their acylglycerol structure, in weaning-piglet diets.

Re-esterified oils are new fat sources obtained from chemical esterification of acid oils with glycerol (both economically interesting by-products from oil refining and biodiesel industries, respectively). The different fatty acid (FA) positional distribution and acylglycerol composition of re-esterified oils may enhance the apparent absorption of saturated fatty acids (SFA) and, thus, their overall nutritive value. The aim of the present study was to investigate the potential use of re-esterified palm oils, in comparison with their corresponding acid and native oils, and also with an unsaturated fat source in weaning-piglet diets. The parameters assessed were: FA apparent absorption, acylglycerol and free fatty acid (FFA) composition of feces, and growth performance. One-hundred and twenty weaning piglets (average weight of 8.50±1.778 kg) were blocked by initial BW (six blocks) and randomly assigned to five dietary treatments, resulting in four piglets per pen and six replicates per treatment. Dietary treatments were a basal diet supplemented with 10% (as-fed basis) of native soybean oil (SN), native palm oil (PN), acid palm oil (PA), re-esterified palm oil low in mono- (MAG) and diacylglycerols (DAG) (PEL), or re-esterified palm oil high in MAG and DAG (PEH). Results from the digestibility balance showed that SN reached the greatest total FA apparent absorption, and statistically different from PN, PA and PEL (P0.05), but PEH achieved the greatest total FA apparent absorption. Animals fed PEL, despite the fact that PEL oil contained more sn-2 SFA, did not show an improved absorption of SFA (P>0.05). Animals fed PA and PN showed similar apparent absorption coefficients (P>0.05), despite the high FFA content of PA oil. The acylglycerol and FFA composition of feces was mainly composed of FFA. There were no significant differences in growth performance (P>0.05). Results of the present study suggest that, despite the different acylglycerol structure of re-esterified oils, there were no significant differences in digestibility or performance with respect to their corresponding PN and PA oils in weaning-piglet diets.

Re-esterified palm oils, compared to native palm oil, do not alter fat absorption, postprandial lipemia or growth performance in broiler chicks.

Re-esterified palm oils are obtained from the chemical esterification of palm acid oils (rich in free fatty acids) with glycerol, both economically interesting by-products from oil refining and biodiesel industries, respectively. Thus, re-esterified palm oils could be an economically interesting alternative to native palm oil in broiler chick diets. However, because they may have different physicochemical properties than have their corresponding native oil, we assessed the effect of fatty acid (FA) positional distribution within acylglycerol molecules and the effect of acylglycerol composition on FA apparent absorption, and their possible consequences on the evolution of postprandial lipemia and growth performance in broiler chicks. Seventy-two 1-day-old female broiler chicks were randomly distributed into 18 cages. The three treatments used were the result of a basal diet supplemented with 6 wt% of native palm oil (N-TAG), re-esterified palm oil (E-TAG), or re-esterified palm oil high in mono- and diacylglycerols (E-MDAG). Chemical esterification raised the fraction of palmitic acid at the sn-2 position from 9.63 mol% in N-TAG oil to 17.9 mol% in E-TAG oil. Furthermore, E-MDAG oil presented a high proportion of mono- (23.1 wt%) and diacylglycerols (51.2 wt%), with FA mainly located at the sn-1,3 positions, which resulted in a lower gross-energy content and an increased solid-fat index at the chicken's body temperature. However, re-esterified palm oils did not alter fat absorption, postprandial lipemia, or growth performance, compared to native palm oil, so they can be used as alternative fat sources in broiler chick diets.

Association of BRCA1 germline mutations in young onset triple-negative breast cancer (TNBC)

BACKGROUND: BRCA1-associated breast cancers have been associated to a triple-negative phenotype. The prevalence of BRCA1 germline mutations in young onset TNBC based on informativeness of family history has not been reported. PATIENTS AND METHODS: From January 2008 to May 2009 were collected blood and tumor samples from patients with TNBC younger than 50 years and without a family history of breast and ovarian cancer in first- and second-degree relatives. Analysis of BRCA1 germline mutations was made. Age at diagnosis and informativeness of family history (presence of female in first- and second-degree relatives alive until age 45) was collected in all cases. Immunohistochemistry of basal-like features was performed centrally in all available tumors. RESULTS: Seven pathogenic mutations were detected in 92 patients (7.6 %), two of them in patients younger than 35 years (28.6 %) (Fisher's exact test, p = 0.631). Three non-classified variants were detected (3.2 %). Family history was informative in two patients with a pathogenic mutation (28.6 %) and not informative in five (71.4 %) (Fisher's exact test, p = 0.121). Of the seven patients with a pathogenic mutation, four had a basal-like phenotype. CONCLUSION: Patients with apparently sporadic TNBC younger than 50 years and a non-informative family history are candidates for germline genetic testing of BRCA1 (AU)

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Association of BRCA1 germline mutations in young onset triple-negative breast cancer (TNBC).

BACKGROUND: BRCA1-associated breast cancers have been associated to a triple-negative phenotype. The prevalence of BRCA1 germline mutations in young onset TNBC based on informativeness of family history has not been reported. PATIENTS AND METHODS: From January 2008 to May 2009 were collected blood and tumor samples from patients with TNBC younger than 50 years and without a family history of breast and ovarian cancer in first- and second-degree relatives. Analysis of BRCA1 germline mutations was made. Age at diagnosis and informativeness of family history (presence of female in first- and second-degree relatives alive until age 45) was collected in all cases. Immunohistochemistry of basal-like features was performed centrally in all available tumors. RESULTS: Seven pathogenic mutations were detected in 92 patients (7.6 %), two of them in patients younger than 35 years (28.6 %) (Fisher's exact test, p = 0.631). Three non-classified variants were detected (3.2 %). Family history was informative in two patients with a pathogenic mutation (28.6 %) and not informative in five (71.4 %) (Fisher's exact test, p = 0.121). Of the seven patients with a pathogenic mutation, four had a basal-like phenotype. CONCLUSION: Patients with apparently sporadic TNBC younger than 50 years and a non-informative family history are candidates for germline genetic testing of BRCA1.

Induced cell toxicity originates dendritic cell death following magnetic hyperthermia treatment.

Magnetic hyperthermia (MH) is based on the use of magnetic nanoparticles (MNPs) to selectively increase the temperature of MNP-loaded target tissues when applying an alternating magnetic field (AMF) in the range of radiofrequency. To date, all MH research has focused on heat generation in an attempt to elucidate the mechanisms for the death of MNP-loaded cells submitted to AMF. However, recent in vitro studies have demonstrated the feasibility of inducing dramatic cell death without increasing the macroscopic temperature during AMF exposure. Here, we show that the cell death observed following AMF exposure, specifically that of MNP-loaded dendritic cells (DCs) in culture, was caused by the release of toxic agents into the cell culture supernatants and not due to a macroscopic temperature increase. We performed MH in vitro experiments to demonstrate that the supernatant of the cell culture following AMF exposure was highly toxic when added to control unloaded DCs, as this treatment led to nearly 100% cell death. Therefore, our results demonstrate that heat is not the only agent responsible for triggering cell death following MH treatment. This finding offers new perspectives for the use of DCs as the proverbial Trojan horse to vectorise MNPs to the target tumour area and these results further support the use of DCs as therapeutic agents against cancer when submitted to AMF. Furthermore, this discovery may help in understanding the mechanism of cell death mediated by exposure to AMF.

Use of recovered frying oils in chicken and rabbit feeds: effect on the fatty acid and tocol composition and on the oxidation levels of meat, liver and plasma.

The addition of some fat co- and by-products to feeds is usual nowadays; however, the regulations of their use are not always clear and vary between countries. For instance, the use of recycled cooking oils is not allowed in the European Union, but they are used in other countries. However, oils recovered from industrial frying processes could show satisfactory quality for this purpose. Here we studied the effects of including oils recovered from the frying industry in rabbit and chicken feeds (at 30 and 60 g/kg, respectively) on the fatty acid (FA) and tocol (tocopherol + tocotrienol) compositon of meat, liver and plasma, and on their oxidative stability. Three dietary treatments (replicated eight times) were compared: fresh non-used oil (LOX); oil discarded from the frying industry, having a high content of secondary oxidation compounds (HOX); and an intermediate level (MOX) obtained by mixing 50 : 50 of LOX and HOX. The FA composition of oil diets and tissues was assessed by GC, their tocol content by HPLC, the thiobarbituric acid value was used to assess tissue oxidation status, and the ferrous oxidation-xylenol orange method was used to assess the susceptibility of tissues to oxidation. Our results indicate that FA composition of rabbit and chicken meat, liver and plasma was scarcely altered by the addition of recovered frying oils to feed. Differences were encountered in the FA composition between species, which might be attributed mainly to differences in the FA digestion, absorption and metabolism between species, and to some physiological dietary factors (i.e. coprophagy in rabbits that involves fermentation with FA structure modification). The α-tocopherol (αT) content of tissues was reduced in response to the lower αT content in the recovered frying oil. Differences in the content of other tocols were encountered between chickens and rabbits, which might be attributable to the different tocol composition of their feeds, as well as to species differences in the digestion and metabolism of tocols. Tissue oxidation and susceptibility to oxidation were in general low and were not greatly affected by the degree of oxidation of the oil added to the feeds. The relative content of polyunsaturated fatty acids/αT in these types of samples would explain the differences observed between species in the susceptibility of each tissue to oxidation. According to our results, oils recovered from the frying industry could be useful for feed uses.

Geographical provenance of palm oil by fatty acid and volatile compound fingerprinting techniques.

Analytical methods are required in addition to administrative controls to verify the geographical origin of vegetable oils such as palm oil in an objective manner. In this study the application of fatty acid and volatile organic compound fingerprinting in combination with chemometrics have been applied to verify the geographical origin of crude palm oil (continental scale). For this purpose 94 crude palm oil samples were collected from South East Asia (55), South America (11) and Africa (28). Partial least squares discriminant analysis (PLS-DA) was used to develop a hierarchical classification model by combining two consecutive binary PLS-DA models. First, a PLS-DA model was built to distinguish South East Asian from non-South East Asian palm oil samples. Then a second model was developed, only for the non-Asian samples, to discriminate African from South American crude palm oil. Models were externally validated by using them to predict the identity of new authentic samples. The fatty acid fingerprinting model revealed three misclassified samples. The volatile compound fingerprinting models showed an 88%, 100% and 100% accuracy for the South East Asian, African and American class, respectively. The verification of the geographical origin of crude palm oil is feasible by fatty acid and volatile compound fingerprinting. Further research is required to further validate the approach and to increase its spatial specificity to country/province scale.

Authentication of organic feed by near-infrared spectroscopy combined with chemometrics: a feasibility study.

Organic products tend to retail at a higher price than their conventional counterparts, which makes them susceptible to fraud. In this study we evaluate the application of near-infrared spectroscopy (NIRS) as a rapid, cost-effective method to verify the organic identity of feed for laying hens. For this purpose a total of 36 organic and 60 conventional feed samples from The Netherlands were measured by NIRS. A binary classification model (organic vs conventional feed) was developed using partial least squares discriminant analysis. Models were developed using five different data preprocessing techniques, which were externally validated by a stratified random resampling strategy using 1000 realizations. Spectral regions related to the protein and fat content were among the most important ones for the classification model. The models based on data preprocessed using direct orthogonal signal correction (DOSC), standard normal variate (SNV), and first and second derivatives provided the most successful results in terms of median sensitivity (0.91 in external validation) and median specificity (1.00 for external validation of SNV models and 0.94 for DOSC and first and second derivative models). A previously developed model, which was based on fatty acid fingerprinting of the same set of feed samples, provided a higher sensitivity (1.00). This shows that the NIRS-based approach provides a rapid and low-cost screening tool, whereas the fatty acid fingerprinting model can be used for further confirmation of the organic identity of feed samples for laying hens. These methods provide additional assurance to the administrative controls currently conducted in the organic feed sector.

Use of palm-oil by-products in chicken and rabbit feeds: effect on the fatty acid and tocol composition of meat, liver and plasma.

This study was undertaken in the framework of a larger European project dealing with the characterization of fat co- and by-products from the food chain, available for feed uses. In this study, we compare the effects, on the fatty acid (FA) and tocol composition of chicken and rabbit tissues, of the addition to feeds of a palm fatty acid distillate, very low in trans fatty acids (TFA), and two levels of the corresponding hydrogenated by-product, containing intermediate and high levels of TFA. Thus, the experimental design included three treatments, formulated for each species, containing the three levels of TFA defined above. Obviously, due to the use of hydrogenated fats, the levels of saturated fatty acids (SFA) show clear differences between the three dietary treatments. The results show that diets high in TFA (76 g/kg fat) compared with those low in TFA (4.4 g/kg fat) led to a lower content of tocopherols and tocotrienols in tissues, although these differences were not always statistically significant, and show a different pattern for rabbit and chicken. The TFA content in meat, liver and plasma increased from low-to-high TFA feeds in both chicken and rabbit. However, the transfer ratios from feed were not proportional to the TFA levels in feeds, reflecting certain differences according to the animal species. Moreover, feeds containing fats higher in TFA induced significant changes in tissue SFA, monounsaturated fatty acids and polyunsaturated fatty acids composition, but different patterns can be described for chicken and rabbit and for each type of tissue.