Algoritmos para la detección de aceite de avellana en aceite de oliva

7 pages, 5 figures, 5 tables.[EN] The fraudulent addition of hazelnut oil to olive oil can be only detected in high proportions (20-25%) using the A-stigmastenol and the difference between triacylglycerols of equivalent carbon number 42, determined experimentally by HPLC and calculated theoretically from the fatty acid composition (AECN42). A new method lies on a sequential comparison of the values of several algorithms with a database built with data obtained from genuine virgin olive oils. The algorithms are: LLLexp en función de %L; (LLL/OLLn)exp - (LLL/OLLn)teor, vs AECN44 and (ECN441LLL), vs %L; being: LLLnexpO LLnexpa nd ECN44ew the percentage of triacylglycerols determined by HPLC, LLLth, OLLnthWr and EcN44th,, the percentage of those calculated theoretically from the fatty acid wmposition; AECN44 the difference between the experimental and theoretica~v alue of ECN44; and finally %L the percentage of linoleic acid. The database has been built considering the values obtained from olive oils of diierent fatty acid composition and from admixtures between them. The method allows the detection of low percentages of hazelnut oil in olive oii ( 5%).[ES] La adición fraudulenta de aceite de avellana en aceite de oliva puede ser detectada sólo en altas proporciones (20–25 %), usando el D7-estigmastenol y la diferencia entre los triglicéridos con número de carbono equivalente igual a 42, determinados experimentalmente por HPLC y teóricamente a partir de la composición de ácidos grasos (DECN42). Se propone un nuevo método que consiste en la comparación de los valores de varios algoritmos con una base de datos de valores experimentales obtenidos de aceites de oliva virgen genuinos. Estos algoritmos son: LLLexp en función de %L; (LLL/OLLn)exp - (LLL/OLLn)teor en función de DECN44 y (ECN44/LLL)exp en función de %L; siendo LLLexp, OLLnexp, y ECN44exp los porcentajes de los triglicéridos obtenidos por HPLC; LLLteor , OLLnteor, y ECN44teor los porcentajes de éstos calculados teóricamente a partir de la composición de ácidos grasos; DECN44 la diferencia entre el valor experimental y teórico del ECN44; y finalmente %L el porcentaje de ácido linoleico. La base de datos se ha recopilado considerando los valores obtenidos de aceites de oliva de diferentes composiciones de ácidos grasos y mezclas entre ellos. El método permite la detección de pequeños porcentajes (5 %) de aceites de avellana en aceites de oliva.We thank to Ministry of Agriculture for the financia1 support (INIA CA098-005).Peer reviewe

Antiangiogenic properties of a nutrient mixture in a model of hemangioma

The pathogenesis of hemangiomas is still largely unknown and the current therapy, such as systemic corticosteroid, vincristine, and interferon-alpha, is toxic and remains unsatisfactory. A nutrient mixture (NM) containing lysine, proline, ascorbic acid and green tea extract has shown significant anti-angiogenic and anti-tumor effect against a number of cancer cell lines. Aim: Using a mouse hemangioendothelioma model, we investigated the efficacy of NM. We also tested the effect of NM in vitro, evaluating cell viability, MMP secretion, invasion, morphology and apoptosis. Methods: Athymic nude mice, 5–6 weeks old, were inoculated with 3 x106 EOMA cells subcutaneously and randomly divided into two groups; group A was fed a regular diet and group B — a regular diet supplemented with 0.5% NM. Four weeks later, the mice were sacrificed and their tumors were excised, weighed and processed for histology. We also tested the effect of NM in vitro. Results: NM inhibited the growth of tumors by 50%. In vitro, NM exhibited dose response cytotoxicity with 10%, 30% and 55% at 10, 100 and 1000 μg/ml. Invasion through Matrigel was inhibited at 50, 100 and 500 μg/ml by 25%, 30% and 100% respectively. NM induced dose-dependent apoptosis of EOMA cells. Conclusions: These results suggest that NM may have therapeutic potential in treating infantile hemangioendotheliomas and, perhaps, other cutaneous vascular tumors

In vivo and in vitro antitumor effects of nutrient mixture in murine leukemia cell line P-388

Aim: Leukemia is characterized by uncontrolled marrow cell proliferation and metastatic foci. We investigated the antitumor potential of a nutrient mixture on malignant leukemia P-388 cells. Methods: The nutrient mixture containing lysine, proline, ascorbic acid, green tea extract and other nutrients is formulated to target key pathways in cancer progression. The cells were treated with the mixture, and tested at doses 0, 10, 50, 100, 500 and 1000 μg/ml in triplicates. The effects were evaluated by cell proliferation, Matrigel invasion, cell morphology and apoptosis. The in vivo effect was measured in male nude mice (n = 12) inoculated with P-388 cells. After randomly dividing in two groups, each group was fed regular and the nutrient mixture supplemented diet and the mice were sacrificed after four weeks. Results: The nutrient mixture decreased P-388 cell proliferation at 500 and 1000 μg/ml. Only 10% cells were viable at 1000 μg/ml. Matrigel invasion was significantly inhibited in a dose dependent manner with virtually total inhibition at 1000 μg/ml. Cell morphological features notably changed with dose increase to 1000 μg/ml. Analysis of apoptotic cells on live green caspase kit exhibited gradual increase with the increasing dose of the nutrient mixture, and at 1000 μg/ml 92% of P-388 cells were in late apoptosis. Tumors in the group of mice supplemented with the nutrient mixture had 50% lower weight compared to the tumors in control group (p = 0.0105). Histopathologically, both the groups of tumors were similar, yet size of tumors in the group treated with the nutrient mixture was considerably smaller. Conclusion: These results indicate that the nutrient mixture exhibited significant action against multiple targets in P-388 leukemia and may have potential in human leukemia

Repression of matrix metalloproteinases and inhibition of cell invasion by a nutrient mixture, containing ascorbic acid, lysine, proline, and green tea extract on human fanconi anemia fibroblast cell lines

Aim: Fanconi Anemia, an autosomal recessive disorder, is characterized by chromosomal abnormality leading to birth defects, progressive bone marrow failure, and a high probability of developing malignancy at an early age. Head and neck squamous cell carcinoma and myeloid leukemia are the major causes of cancer related morbidity and mortality in Fanconi anemia patients. Me­thods: We investigated the effect of a nutrient mixture on Fanconi Anemia human fibroblast cell lines FA‐A:PD20 and FA‐A:PD220 on matrix metalloproteinase expression, invasion, cell proliferation, morphology and apoptosis. The cell lines were grown in a modified Dulbecco’s Eagle medium and at near confluence were treated with the nutrient mixture at increasing doses: 0; 10; 50; 100; 500; 1000 µg/ml. The cells were also treated with PMA to induce MMP-9 expression. Results: Zymography demonstrated MMP‐2 and PMA‐induced MMP‐9 activity. The nutrient mixture inhibited expression of both, MMP-2 and MMP-9, in a dose dependent manner with virtually total inhibition observed at 500 µg/ml. Matrigel invasion was inhibited in both cells lines; with 100% inhibition for FA-A:PD20 at 500 µg/ml and 100% inhibition of FA-A:P220 cells at 100 µg/ml. H&E staining did not indicate any change in cell morphology and causes apoptosis at higher doses. Conclusion: Our data demonstrated that the nutrient mixture inhibited matrix metalloproteinase expression, invasion and induced apoptosis, the important parameters for cancer prevention. The results suggest that the nutrient mixture may have therapeutic potential in Fanconi Anemia associated neoplasia