Carine Ferrand

Proton NMR quantitative profiling for quality assessment of greenhouse-grown tomato fruit

Tomato is an essential crop in terms of economic importance and nutritional quality. In France, the third most important region for tomato (Solanum lycopersicum L.) production is Aquitaine where the major part of production is now grown soilless under greenhouse conditions with harvest from March to November. Tomato fruit quality at harvest is a direct function of its metabolite content at that time. The aim of this work was to use a global approach to characterize changes in the fruit organoleptic quality at harvest under commercial culture conditions during an entire season for two varieties and two different fertilization practices (with or without recycling of the nutrient solution) for one variety. Absolute quantification data of 32 major compounds in fruit without seeds were obtained through untargeted (proton nuclear magnetic resonance, 1H-NMR) quantitative profiling. These data were complemented by colorimetric analysis of ascorbate and total phenolics. They were analyzed with chemometric approaches. Principal component analysis (PCA) or partial least square analyses (PLS) revealed more discriminant metabolites for season than for variety and showed that nutrient solution recycling had very little effect on fruit composition. These tendencies were confirmed with univariate analyses. 1H-NMR profiling complemented with colorimetric analyses therefore provided a diagnostic tool to follow the changes in organoleptic and nutritional quality of tomato. In addition the quantitative information generated will help to increase our knowledge on the mechanisms of plant response to environmental modifications.

Effect of tyramine, a dietary amine, on glycerol and lactate release by isolated adipocytes from old rats.

Amine degradation by adipocyte amine oxidases leads to the production of metabolites that interact with lipid and glucose metabolisms and their hormonal regulations. To further investigate these interactions, we determined the effect of a dietary amine, tyramine (TYR), on glycerol and lactate releases, respectively taken as indices of lipolytic and glycolytic activities of isolated adipocytes. Old male Wistar rats were used to prepare adipocytes by collagenase dissociation of retroperitoneal fat pads. The two tested doses of tyramine (10 μM and 1 mM) had no effect on basal glycerol release. On the other hand, TYR, at the highest dose tested (1 mM), weakly but significantly increased basal lactate release, which was elevated in adipocytes from old rats. Norepinephrine (NE), highly stimulated adipocyte lipolysis with a submaximal effect at 1 μM which was slightly but significantly inhibited by TYR 1 mM. Insulin 1 nM (INS) also poorly inhibited the NE-stimulated lipolysis in adipocytes isolated from old rats. TYR was able to potentiate the poor antilipolytic efficiency of INS. Under similar conditions, a high dose of NE greatly reduced lactate production and TYR (1 mM) reversed this inhibition of lactate release. INS was also able to totally reverse the inhibitory effect of NE on lactate release, but there was no potentiation between insulin and tyramine effects. It can be concluded that high doses of TYR interact with norepinephrine and insulin, at least on the control of glycerol and lactate release, by counteracting catecholamine effects and by mimicking insulin actions.ResumenLa degradación de aminas por las aminooxidasas del tejido adiposo produce metabolitos capaces de influir sobre los metabolismos lipídico y glucídico y su regulación hormonal. Para investigar acerca de estos efectos, se ha estudiado el efecto de la tiramina (TIR), una amina presente en la dieta, sobre la producción de glicerol y de lactato, como índice respectivo de la lipolisis y de la glicolisis. A partir de tejido adiposo perirrenal de machos viejos de rata Wistar se aislaron adipocitos tras digestión con colagenasa. Se estudióin vitro el efecto de la tiramina a dos concentraciones, 10 μM y 1 mM, sobre la liberación de glicerol y de lactato. Ninguna de ellas modificó la liberación basal de glicerol, pero la mayor estimuló la liberación basal de lactato, ya bastante elevada. La norepinefrina (NE) estimuló fuertemente la lipolisis con un efecto submáximo a 1 μM, que fué inhibido parcial pero significativamente por TIR 1 mM. La producción basal de lactato se redujo por dosis elevadas de NE y esta inhibición se anuló por adición TIR 1 mM. Ademas, la tiramina fue capaz de reforzar la débil acción antilipolítica de la insulina en los adipocitos antilipolítica de la insulina en los adipocitos estimulados por norepinefrina de ratas viejas. Por otra parte, la insulina anuló el efecto inhibidor de la NE sobre la producción basal de lactato sin que se observara potenciación por la presencia de TIR. Estos resultados parecen indicar que la tiramina, a dosis elevadas, puede reemplazar parcialmente el efecto de la insulina e influir sobre el control de la liberación de glicerol y de lactato por la norepinefrina

Synergic effect of vitamin A and high-fat diet in adipose tissue development and nuclear receptor expression in young rats

In order to study the effects of dietary lipids and vitamin A on the development of adipose tissues, young rats were submitted for 8 d to a control or to two cafeteria diets with normal (Caf) or higher (Caf + ) vitamin A levels. Retinoid (retinoic acid receptor (RAR) a, RARg, retinoid X receptor(RXR) alpha) and fatty acid (PPARgamma) receptor mRNA was measured in the subcutaneous white adipose tissue (Swat) and in isolated mature adipocytes by RT-PCR. The stroma vascular fraction was cultured in vitro to test the capacities of the adipocyte precursors to proliferate and differentiate.The Caf diet enriched in vitamin A resulted in an increased adiposity, due to increased adipocyte hypertrophy. This was concomitant with a lower expression of RARa and RARg mRNA (234.6 and 238.6 %) and a higher expression of PPARgamma (+59 %) in the Swat and, to a less extent,in isolated adipocytes. Positive correlations were obtained between PPARgamma mRNA and Swat weights and between PPARgamma and RXRalpha mRNA. By contrast, RARgamma mRNA and Swat masses were negatively correlated. The adipocyte precursors from Caf + Swat proliferated more,in vitro, at the beginning of the culture. This difference progressively disappeared and was totally absent after 8 d of culture, but with a higher percentage of differentiated preadipocytes (+80.3 %) in the Caf + group. In conclusion, lipids and vitamin A act synergistically on the normal growth of the adipose tissue in young rats, concomitant with an imbalance in the pattern of the nuclear receptors. These changes influence the early normal development of the endogenous adipocyte precursors.

Effect of vitamin A content in cafeteria diet on the expression of nuclear receptors in rat subcutaneous adipose tissue

The aim of this study was to determine the effects of cafeteria diet containing control or elevated level of vitamin A on the expression of nuclear receptors in adipose tissue. Male Wistar rats were submitted to 3 experimental diets during 8 weeks, a standard diet and two hyper-energetic, hyperlipidic “cafeteria” diets containing normal (Caf) or higher (Caf+) vitamin A level. During the experiment, body weights and energy intakes were measured. At the end of the experimental period, subcutaneous adipose tissue (Swat) and all the fat mass were removed and weighted. Nuclear receptors mRNA levels of RARα, RARγ, RXRα, PPARγ were measured in the Swat by a real-time semi-quantitative RT-PCR method. We observed that energy intake of Caf+ and Caf groups was significantly higher than that of the control group. Despite a higher increase of the energy intake in the Caf group compared to the Caf+ group, no significant difference was observed in the body weight gain of the Caf+ compared to the Caf group. The Caf+ and Caf diets led to a significant increase of adipose tissue in cafeteria groups as observed in the Swat depot. The mRNA levels of PPARγ and RXRα were significantly increased in the Caf+ group as compared to control group, with a significant positive correlation between these two parameters. Expressions of RARα and RARγ were not modified in experimental groups compared to controls. In conclusion, 8-week exposure to cafeteria diets with normal and higher levels of vitamin A led to an increase of adiposity in rats, associated, only in the group fed with the higher vitamin A level cafeteria diet, with an increase of PPARγ and RXRα expressions in subcutaneous adipose tissue.ResumenEl objetivo del presente trabajo consiste en determinar las consecuencias de un alto contenido en vitamina A en dieta de cafetería sobre la expresión de receptores nucleares en el tejido adiposo. Así, ratas macho Wistar se dividieron en tres grupos: Durante 8 semanas, el grupo control se alimentó con pienso estándar, mientras que los grupos tratados recibieron una dieta rica en grasa (dieta de cafetería) enriquecida (Caf+) o no (Caf) con vitamina A. El peso corporal y la ingesta se determinaron durante todo el experimento. Al final del tratamiento, se pesó el tejido adiposo subcutáneo (Swat) y las otras reservas de grasa. Los niveles de ARNm de los receptores nucleares RARα, RARγ, RXRα, PPARγ se determinaron en el Swat con un método semi-cuantitativo de RT-PCR en tiempo real. Las ingestas energéticas de los grupos Caf+ y Caf fueron significativamente mayores que las del grupo control. A pesar del aumento en la ingesta del grupo Caf respecto del Caf+, no se observaron diferencias significativas en el aumento de peso corporal entre ambos grupos. Además, las dietas de los grupos Caf+ y Caf provocaron un claro aumento del tamaño de las reservas de grasa, incluido el peso del Swat. Los niveles de ARNm de PPARγ y RXRα se incrementaron significativamente en el grupo Caf+ respecto del control, con correlación positiva entre ambos. En cambio, no se modificó la expresión de RARα y RARγ. En suma, 8 semanas de alimentación con dieta de cafetería con niveles normales o elevados de vitamina A dan lugar a aumento de la adiposidad en la rata, asociado con aumento de la expresión de PPARγ y RXRα en el tejido adiposo subcutáneo solo en el grupo que recibió suplemento de vitamina A.

Prolonged treatment with the β3-adrenergic agonist CL 316243 induces adipose tissue remodeling in rat but not in guinea pig: 1) fat store depletion and desensitization of β-adrenergic responses

Abstractβ3-adrenergic agonists have been considered as potent antiobesity and antidiabetic agents mainly on the basis of their beneficial actions discovered twenty years ago in obese and diabetic rodents. The aim of this work was to verify whether prolonged treatment with a β3-adrenergic agonist known to stimulate lipid mobilisation, could promote desensitization of β-adrenergic responses. Wistar rats and guinea pigs were treated during one week with CL 316243 (CL, 1 mg/kg/d) by implanted osmotic minipumps. In control animals, β3-adrenergic agonists were lipolytic in rat but not in guinea pig adipocytes. CL-treatment did not alter body weight gain in both species, but reduced fat stores in rats. Lipolysis stimulation by forskolin was unmodified but responses to β1-, β2- and β3-agonists were reduced in visceral or subcutaneous white adipose tissues of CL-treated rats. Similarly, the β3-adrenergic-dependent impairment of insulin action on glucose transport and lipogenesis in rat adipocytes was diminished after CL-treatment. In rat adipocytes, [125I]ICYP binding and β3-adrenoceptor mRNA levels were reduced after sustained CL administration. These findings show that CL 316243 exerts β3-adrenergic lipolytic and antilipogenic effects in rat adipocytes. These actions, which are likely involved in the fat depletion observed in rat, also lead to the desensitization of all β-adrenergic responses. Therefore this desensitization, together with the lack of slimming action in guinea pig, seriously attenuates the usefulness of β3-agonists as antiobesity agents, and may explain why such agonists have not been conducted to a widespread clinical, use.ResumenLos agonistas β3-adrenérgicos son considerados potentes agentes anti-obesidad y antidiabéticos debido, fundamentalmente, a los efectos beneficiosos que producen en roedores obesos y diabeticos, descubiertos ya hace veinte años. El objetivo del presente estudio fue verificar si un tratamiento prolongado con agonists β3-adrenérgicos, conocidos estimulantes de la movilización lipídica, puede promover la desensibilización de las respuestas β-adrenérgicas. Para ello, se trataron ratas Wistar y cobayas con CL 316243 (CL, 1 mg/kg/d), administrado mediante el implante de minibombas osmóticas, durante una semana. En los adipocitos de ratas control, pero no en los de cobayas control, los agonistas β3-adrenérgicos produjeron efectos lipolíticos. El tratamiento con CL no modificó la ganancia de peso en ninguna de las dos especies, pero redujo los depósitos de grasa en ratas. En el tejido adiposo visceral y subcutáneo de las ratas tratadas con CL, la estimulación de la lipólisis por forskolina no se vió afectada, pero las respuestas a agonistas β1, β2, y β3 se redujeron. De manera análoga, el deterioro de la función insulínica, en lo que al transporte de glucosa y la lipogénesis se refiere, producido por los adrenérgicos β3 y que sólo se observa en los adipocitos de rata, disminuyó tras el tratamiento con CL. En los adipocitos de rata, la unión a [125I]ICYP y los niveles de ARNm del receptor adrenérgico β3 disminuyeron con la administración sostenida de CL. Estos resultados demuestran que el CL 316243 produce efectos lipolítico y antilipogénico únicamente en los adipocitos de rata. Estas acciones, muy probablemente relacionadas con la depleción de grasa observada en la rata, conducen a la desensibilización de todas las respuestas β-adrenérgicas. Esta desensibilización, junto con la ausencia de efecto adelgazante en cobayas, reduce la utilidad de los agonistas β3-adrenérgicos como agentes antiobesidad y podría explicar por qué no se han utilizado en la práctica clínica habitual como nuevos fármacos.

Sorbic acid-amine function interactions.

Sorbic acid has a system of conjugated double bonds which makes it able to undergo nucleophilic addition reactions with certain functions. The interactions between sorbic acid and amine functions present in the endogenous constituents of food were quantified. The formation of new products was demonstrated and the underlying mechanisms studied using ethyl sorbate and various amines. HPLC, GC, GC-SM and NMR analyses of the reaction mixtures enabled the products to be isolated and identified. The addition reactions led, at 20 degrees C, to linear monoadducts and, at 50 degrees C and 80 degrees C, to cyclic derivatives resulting from double addition.

Influence of various parameters on the browning of potassium sorbate in the presence of amines

Potassium sorbate can undergo oxidation to form carbonyl moieties and cause browning. This investigation studied the fate of the compounds produced by auto-oxidation of potassium sorbate and measured the browning in the presence of amines. Experimental plans in which four factors were combined (temperature, oxygen, amine and light) led to the observation that the interaction between oxygen and high temperature (75°C) favoured browning, independently of the presence of amine. At 4°C, the amine seemed to cause a decrease in the proportion of carbonyl-containing compounds which would appear to participate in carbonylamine reactions. At 75°C, the amine forms adducts with the potassium sorbate. In parallel, high temperature favours auto-oxidation reactions that produce carbonyls. GC/MS and NMR analysis of the reaction products of potassium sorbate/amine mixtures led to the identification of cyclic products.