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Effect of Theobroma cacao flavonoids on immune activation of a lymphoid cell line.

We analysed the effect of (-)-epicatechin and cocoa extract on the activation of a lymphoid cell line. Particularly the expression of IL-2 receptor alpha (IL-2Ralpha or CD25) and, the secretion of IL-2 and IL-4 were established after flavonoid treatment. Two media culture conditions (1 and 10 % of fetal calf serum supplementation) and the different moments of flavonoid addition (simultaneously or 2 h before cell-activation) were compared. IL-2Ralpha (CD25) expression on activated cells was significantly reduced by epicatechin and cocoa extract in a dose-dependent manner, achieving the highest inhibition of about 50 % when flavonoids were added 2 h before stimulation. IL-2 secretion was also inhibited by the presence of both epicatechin and cocoa extract, displaying 60 and 75 % of inhibition, respectively. Cocoa flavonoids were also able to enhance 3-4.5-fold IL-4 release. In summary, cocoa extract down-modulated T lymphocyte activation and therefore the acquired immune response. This fact could be important in some states of the immune system hyperactivity such as autoimmune or chronic inflammatory diseases.

Animals↗

Apparent metabolisable energy and digestibility of shea (Vitellaria paradoxa) fat, cocoa (Theobroma cacao) fat and soybean oil in broiler chicks.

1. The objective of this experiment was to determine and compare the apparent lipid digestibility coefficient and apparent metabolisable energy (AME) value of shea nut (Vitellaria paradoxa, Gaertn.) fat in broiler chickens with that of soybean oil and cocoa fat. 2. One hundred and sixty 13-d-old male broiler chicks were used in a randomised complete block design. The fats were added at 30, 60 and 90 g/kg to a basal diet. A tenth dietary treatment was the basal feed with no added fats or oils. The birds were fed on the diets for 8 d and all droppings were collected for the final 4 d. 3. The mean coefficient of apparent lipid digestibility for shea fat (0.58) was similar to that of cocoa fat (0.54) but lower than that of soybean oil (0.95). There was evidence of a lipid x concentration interaction with the 90 g/kg shea fat diet having low lipid digestibility (0.43). 4. There was an interaction between the effects of dietary lipid concentration and test lipid on AME but, at dietary levels of 60 g/kg and below, the AME of shea fat (22.0 MJ/kg DM) and cocoa fat (26.4 MJ/kg DM) was significantly lower than that of soybean oil (39.8 MJ/kg DM).

Animal Feed↗

Dietary flavanols and procyanidin oligomers from cocoa (Theobroma cacao) inhibit platelet function.

BACKGROUND: Flavonoids may be partly responsible for some health benefits, including antiinflammatory action and a decreased tendency for the blood to clot. An acute dose of flavanols and oligomeric procyanidins from cocoa powder inhibits platelet activation and function over 6 h in humans. OBJECTIVE: This study sought to evaluate whether 28 d of supplementation with cocoa flavanols and related procyanidin oligomers would modulate human platelet reactivity and primary hemostasis and reduce oxidative markers in vivo. DESIGN: Thirty-two healthy subjects were assigned to consume active (234 mg cocoa flavanols and procyanidins/d) or placebo (< or = 6 mg cocoa flavanols and procyanidins/d) tablets in a blinded parallel-designed study. Platelet function was determined by measuring platelet aggregation, ATP release, and expression of activation-dependent platelet antigens by using flow cytometry. Plasma was analyzed for oxidation markers and antioxidant status. RESULTS: Plasma concentrations of epicatechin and catechin in the active group increased by 81% and 28%, respectively, during the intervention period. The active group had significantly lower P selectin expression and significantly lower ADP-induced aggregation and collagen-induced aggregation than did the placebo group. Plasma ascorbic acid concentrations were significantly higher in the active than in the placebo group (P < 0.05), whereas plasma oxidation markers and antioxidant status did not change in either group. CONCLUSIONS: Cocoa flavanol and procyanidin supplementation for 28 d significantly increased plasma epicatechin and catechin concentrations and significantly decreased platelet function. These data support the results of acute studies that used higher doses of cocoa flavanols and procyanidins.

Adult↗

Localization and identification of phenolic compounds in Theobroma cacao L. somatic embryogenesis.

Cocoa breeders and growers continue to face the problem of high heterogeneity between individuals derived from one progeny. Vegetative propagation by somatic embryogenesis could be a way to increase genetic gains in the field. Somatic embryogenesis in cocoa is difficult and this species is considered as recalcitrant. This study was conducted to investigate the phenolic composition of cocoa flowers (the explants used to achieve somatic embryogenesis) and how it changes during the process, by means of histochemistry and conventional chemical techniques. In flowers, all parts contained polyphenolics but their locations were specific to the organ considered. After placing floral explants in vitro, the polyphenolic content was qualitatively modified and maintained in the calli throughout the culture process. Among the new polyphenolics, the three most abundant were isolated and characterized by 1H- and 13C-NMR. They were hydroxycinnamic acid amides: N-trans-caffeoyl-l-DOPA or clovamide, N-trans-p-coumaroyl-l-tyrosine or deoxiclovamide, and N-trans-caffeoyl-l-tyrosine. The same compounds were found also in fresh, unfermented cocoa beans. The synthesis kinetics for these compounds in calli, under different somatic embryogenesis conditions, revealed a higher concentration under non-embryogenic conditions. Given the antioxidant nature of these compounds, they could reflect the stress status of the tissues.

Cacao↗

Genetic differentiation of cocoa (Theobroma cacao L.) populations revealed by RAPD analysis.

In order to preserve and exploit the valuable genetic resources of tropical forest trees, such as cocoa, a systematic assessment of the available genetic variability is necessary. The approach we have used is based on a simple mini-prep DNA extraction procedure together with a polymerase-chain-reaction- (PCR)-based polymorphic assay procedure (RAPD). Twenty-five cocoa accessions: IMCs and PAs collected from Peru and LCTEENs collected from Ecuador, which are difficult to distinguish using morphological or biochemical descriptors, were uniquely fingerprinted using a minimum of three oligonucleotide primers. Analysis of the variability detected using RAPDs clearly discriminated between the geographical origin of the three cocoa populations. Partitioning of variability into within and between population components revealed that most variation was detected within a population. The potential of RAPD analysis to facilitate the rationalization of field gene banks and provide accurate estimates of diversity to allow optimization of collecting strategies is discussed.

Base Sequence↗

Pentameric procyanidins isolated from Theobroma cacao seeds selectively downregulate ErbB2 in human aortic endothelial cells.

Flavonoids isolated from cocoa have biological activities relevant to oxidant defenses, vascular health, tumor suppression, and immune function. The intake of certain dietary flavonoids, along with other dietary substances such as tocopherols, ascorbate, and carotenoids, is epidemiologically associated with a reduced risk of cardiovascular disease. Flavonoids have also been shown to modulate tumor pathology in vitro and in animal models. We took advantage of the conserved sequences found in tyrosine kinases to study the influence of cocoa fractions and controls on gene expression. We report that the pentameric procyanidin (molecular weight of 1442 daltons) fraction isolated from cocoa was a potent inhibitor of tyrosine kinase ErbB2 expression, a receptor important in angiogenesis regulation. Consistent with this primary observation, the cocoa flavonoid fraction also suppressed human aortic endothelial cell (HAEC) growth and decreased expression of two tyrosine kinases responsive to ErbB2 modulation, namely VEGFR-2/KDR and MapK 11/p38beta2. These inhibitory effects were observed when HAECs were treated with the flavonol fraction (molecular weight 280 daltons) isolated from cocoa, which comprise the structural subunits from which the procyanidin flavonoid subclass is biosynthetically constructed. Down-regulation of ErbB2 and inhibition of HAEC growth by cocoa procyanidins may have several downstream implications, including reduced vascular endothelial growth factor (VEGF) activity and angiogenic activity associated with tumor pathology. These results suggest specific dietary flavonoids are capable of selectively inhibiting ErbB2 and therefore may offer important insight into the design of therapeutic agents that target tumors overexpressing ErbB2.

Antioxidants↗

Establishment of the fungal entomopathogen Beauveria bassiana (Ascomycota: Hypocreales) as an endophyte in cocoa seedlings (Theobroma cacao).

The fungal entomopathogen Beauveria bassiana became established as an endophyte in in vitro-grown cocoa seedlings tested for up to 2 mo after inoculation to the radicle with B. bassiana suspensions. The fungus was recovered in culture from stems, leaves and roots. B. bassiana also was detected as an epiphyte 1 and 2 mo postinoculation. Penicillium oxalicum and five bacterial morphospecies also were detected, indicating that these were present as endophytes in the seed.

Animals↗

Characterization of cocoa butter extracted from hybrid cultivars of Theobroma cacao L.

Cocoa butter is the most important fat used in the confectionery and chocolate industries. The main objective of the present study was to determine the physical and chemical characteristics of cocoa butter extracted from hybrid cultivars belonging to the germplasm bank of the Fondo Nacional de Investigaciones Agropecuarias (National Foundation for Agricultural Research). AOAC methods were used for the assessment of the proximal composition of the beans, physical and chemical characteristics as well as for the fatty acid profile of the fat. It was found that there were statistical differences in the proximate composition of the cocoa beans among the cultivars studied as well as the iodine and saponification indices of the butter. Saturated fatty acids were present in higher proportions than unsaturated fatty acids, with palmitic and stearic acid as the main fractions. Oleic acid content was higher than linoleic acid. The fatty acid profile found is the main factor that influences the hard texture of the cocoa butter from Venezuelan cocoa hybrids cultivars.

Butter↗

Somatic embryogenesis from flower explants of cocoa (Theobroma cacao L.).

Two types of flower explants, staminoides and petals, were used for in vitro induction of somatic embryos in cocoa. After 14 days in culture, we observed globular structures and callus formation on both types of explants. However, the better results were obtained on staminoides: 98.3% formed callus and 86.2% somatic embryos on Murashige and Skoog (1962) medium supplemented with sucrose, coconut water, 2,4-D, kinetin and agar.

Cacao↗